Proceedings Latest open access articles published in Proceedings at https://www.mdpi.com/journal/proceedings https://www.mdpi.com/journal/proceedings MDPI en Creative Commons Attribution (CC-BY) MDPI support@mdpi.com Proceedings, Vol. 147, Pages 2: Student Engagement, Autonomy and Teacher Support in Romanian High Schools: Evidence from a National Study https://www.mdpi.com/2504-3900/147/1/2 This study situates autonomy-related learning behaviors and forms of teacher support within the broader framework of student engagement in Romanian high schools. It presents a secondary analysis of a national mixed-methods study conducted in 2023. Quantitative analyses used responses from 3181 students in a two-stage cluster survey involving 237 school/specialization units; qualitative evidence came from focus groups and interviews in ten schools. Descriptive analyses, selected correlations, and an individual-level cross-tabulation were integrated with thematic findings. Personal learning methods, self-assessment, strategy adaptation, and a preference for learning independently were frequently reported, whereas learning-time management and additional self-directed learning were less common. Preferences for learning independently and for teacher explanations coexisted among 66.7% of students; teacher explanations were similarly endorsed across groups reporting higher and lower frequencies of learning better independently. Selected planning, monitoring, and strategy-adjustment behaviors were positively associated with corrective feedback, teacher explanations, or teacher-selected resources. Qualitative accounts showed how pedagogical guidance may follow independent effort and support understanding and strategy adjustment, while workload, long school days, and limited learning time may constrain responsibility for learning. The findings support guided autonomy as an analytical perspective linking student initiative with pedagogical structure, formative feedback, and relevant resources. 2026-09-08 Proceedings, Vol. 147, Pages 2: Student Engagement, Autonomy and Teacher Support in Romanian High Schools: Evidence from a National Study

Proceedings doi: 10.3390/proceedings2026147002

Authors: Adela M. Țăranu Simona A. Pascal

This study situates autonomy-related learning behaviors and forms of teacher support within the broader framework of student engagement in Romanian high schools. It presents a secondary analysis of a national mixed-methods study conducted in 2023. Quantitative analyses used responses from 3181 students in a two-stage cluster survey involving 237 school/specialization units; qualitative evidence came from focus groups and interviews in ten schools. Descriptive analyses, selected correlations, and an individual-level cross-tabulation were integrated with thematic findings. Personal learning methods, self-assessment, strategy adaptation, and a preference for learning independently were frequently reported, whereas learning-time management and additional self-directed learning were less common. Preferences for learning independently and for teacher explanations coexisted among 66.7% of students; teacher explanations were similarly endorsed across groups reporting higher and lower frequencies of learning better independently. Selected planning, monitoring, and strategy-adjustment behaviors were positively associated with corrective feedback, teacher explanations, or teacher-selected resources. Qualitative accounts showed how pedagogical guidance may follow independent effort and support understanding and strategy adjustment, while workload, long school days, and limited learning time may constrain responsibility for learning. The findings support guided autonomy as an analytical perspective linking student initiative with pedagogical structure, formative feedback, and relevant resources.

]]> Student Engagement, Autonomy and Teacher Support in Romanian High Schools: Evidence from a National Study Adela M. Țăranu Simona A. Pascal doi: 10.3390/proceedings2026147002 Proceedings 2026-09-08 Proceedings 2026-09-08 147 1 Proceeding Paper 2 10.3390/proceedings2026147002 https://www.mdpi.com/2504-3900/147/1/2 Proceedings, Vol. 145, Pages 10: Teachers’ Interest, Self-Confidence, and Perceived Ability to Integrate Historical Scientific Instruments into Their Teaching https://www.mdpi.com/2504-3900/145/1/10 This study investigates whether a teacher professional development programme on the educational use of Historical Scientific Instruments enhances teachers’ interest, self-confidence, and perceived ability to integrate HSIs into science teaching. The study was conducted within the Erasmus+ INSIGHT project and involved 25 primary and secondary school teachers participating in a 26 h blended professional development programme; 19 completed a retrospective pre–post questionnaire, and three participated in semi-structured interviews. According to initial findings, participants reported significantly higher post-programme scores across all three dimensions, highlighting the value of inquiry-based, hands-on, and historically contextualized learning experiences in their reported professional learning. 2026-09-02 Proceedings, Vol. 145, Pages 10: Teachers’ Interest, Self-Confidence, and Perceived Ability to Integrate Historical Scientific Instruments into Their Teaching

Proceedings doi: 10.3390/proceedings2026145010

Authors: Constantina Stefanidou Panagiotis (Takis) Lazos Vasiliki Psoma Constantine Skordoulis

This study investigates whether a teacher professional development programme on the educational use of Historical Scientific Instruments enhances teachers’ interest, self-confidence, and perceived ability to integrate HSIs into science teaching. The study was conducted within the Erasmus+ INSIGHT project and involved 25 primary and secondary school teachers participating in a 26 h blended professional development programme; 19 completed a retrospective pre–post questionnaire, and three participated in semi-structured interviews. According to initial findings, participants reported significantly higher post-programme scores across all three dimensions, highlighting the value of inquiry-based, hands-on, and historically contextualized learning experiences in their reported professional learning.

]]> Teachers’ Interest, Self-Confidence, and Perceived Ability to Integrate Historical Scientific Instruments into Their Teaching Constantina Stefanidou Panagiotis (Takis) Lazos Vasiliki Psoma Constantine Skordoulis doi: 10.3390/proceedings2026145010 Proceedings 2026-09-02 Proceedings 2026-09-02 145 1 Proceeding Paper 10 10.3390/proceedings2026145010 https://www.mdpi.com/2504-3900/145/1/10 Proceedings, Vol. 145, Pages 9: Enhancing Creative Thinking Through Coding: Preliminary Evidence from an Experimental Study in Primary and Lower-Secondary Education https://www.mdpi.com/2504-3900/145/1/9 Coding and educational robotics are increasingly promoted in schools as catalysts for creative thinking, yet recent reviews suggest that this link is not automatic and depends critically on pedagogical design. Empirical evidence from Italian compulsory education remains scarce. We report preliminary findings from the Coding&Learning project, a quasi-experimental study involving 291 enrolled students (138 primary, 153 lower-secondary) and 23 trainers across four Italian schools. Verbal divergent thinking was assessed pre- and post-intervention with the Divergent Association Task (DAT); complete pre–post data were available for 217 students. All four groups improved between pre- and post-test, and within-group gains were statistically significant in both experimental groups (primary: +3.27 points, p = 0.031; lower-secondary: +2.61 points, p = 0.007) and in the lower-secondary control group. When experimental and control classes were compared directly—through gain-score t-tests and ANCOVA controlling for pre-test—the between-condition differences did not reach significance, and effect sizes were small (primary g = 0.18; lower-secondary g ≈ 0), as is not unexpected for a brief, statistically underpowered preliminary study. At the primary level, a larger share of experimental students improved (66.7% vs. 52.3%). Trainers reported coherent perceived gains in dispositions adjacent to verbal divergent thinking. Findings are specific to verbal divergent thinking and should not be generalised to creativity as a broader construct. 2026-09-02 Proceedings, Vol. 145, Pages 9: Enhancing Creative Thinking Through Coding: Preliminary Evidence from an Experimental Study in Primary and Lower-Secondary Education

Proceedings doi: 10.3390/proceedings2026145009

Authors: Federica Pelizzari Marta Giudici Michele Marangi Simona Ferrari

Coding and educational robotics are increasingly promoted in schools as catalysts for creative thinking, yet recent reviews suggest that this link is not automatic and depends critically on pedagogical design. Empirical evidence from Italian compulsory education remains scarce. We report preliminary findings from the Coding&Learning project, a quasi-experimental study involving 291 enrolled students (138 primary, 153 lower-secondary) and 23 trainers across four Italian schools. Verbal divergent thinking was assessed pre- and post-intervention with the Divergent Association Task (DAT); complete pre–post data were available for 217 students. All four groups improved between pre- and post-test, and within-group gains were statistically significant in both experimental groups (primary: +3.27 points, p = 0.031; lower-secondary: +2.61 points, p = 0.007) and in the lower-secondary control group. When experimental and control classes were compared directly—through gain-score t-tests and ANCOVA controlling for pre-test—the between-condition differences did not reach significance, and effect sizes were small (primary g = 0.18; lower-secondary g ≈ 0), as is not unexpected for a brief, statistically underpowered preliminary study. At the primary level, a larger share of experimental students improved (66.7% vs. 52.3%). Trainers reported coherent perceived gains in dispositions adjacent to verbal divergent thinking. Findings are specific to verbal divergent thinking and should not be generalised to creativity as a broader construct.

]]> Enhancing Creative Thinking Through Coding: Preliminary Evidence from an Experimental Study in Primary and Lower-Secondary Education Federica Pelizzari Marta Giudici Michele Marangi Simona Ferrari doi: 10.3390/proceedings2026145009 Proceedings 2026-09-02 Proceedings 2026-09-02 145 1 Proceeding Paper 9 10.3390/proceedings2026145009 https://www.mdpi.com/2504-3900/145/1/9 Proceedings, Vol. 150, Pages 13: Should Biological Markers Be Included in Forensic Psychiatric Reports? https://www.mdpi.com/2504-3900/150/1/13 Background: Across jurisdictions, forensic psychiatry evaluations primarily rely on clinical assessment, psychometric testing, and historical and behavioural information. Following advances in neuroscience and neurolaw, there is a growing interest in incorporating biological markers (such as structural and functional neuroimaging (structural MRI, CT, fMRI), electrophysiology (EEG)) into forensic psychiatric reports. Empirical studies indicate that courts are increasingly encountering such data, particularly in cases of homicide, severe violence, insanity evaluations, and capital sentencing. Methods/Results: Neuroimaging can document brain disease or injury that may impair cognition, affect regulation, or impulse control, important to assessment of criminal accountability, and future risk. Biological markers may corroborate psychiatric and neurological diagnoses, and as such make expert reasoning more transparent. Assessments that omit biological investigations may be considered incomplete, with a risk of undermining fair judicial process and equality before the law, as well as hampering future treatment and risk management. However, there are notable challenges that limit the use of biomarkers in forensic psychiatry evaluations. Neuroscientific findings have methodological and inferential limitations affecting their practical use in individual cases. Furthermore, there is a mismatch between the empirical nature of neuroscience and the normative legal questions such as those of criminal accountability and intent. Additional concerns include risks of over-interpretation (“brain overclaim”), “double-edged sword” effects (mitigation versus inferred dangerousness), inequitable access, and the absence of robust, jurisdiction-specific standards. Conclusions: The inclusion of biological markers in forensic psychiatry evaluations has considerable potential benefits but also raises challenges that need to be addressed in future practice, research, and guideline development. 2026-09-01 Proceedings, Vol. 150, Pages 13: Should Biological Markers Be Included in Forensic Psychiatric Reports?

Proceedings doi: 10.3390/proceedings2026150013

Authors: Unn Kristin Hauvik

Background: Across jurisdictions, forensic psychiatry evaluations primarily rely on clinical assessment, psychometric testing, and historical and behavioural information. Following advances in neuroscience and neurolaw, there is a growing interest in incorporating biological markers (such as structural and functional neuroimaging (structural MRI, CT, fMRI), electrophysiology (EEG)) into forensic psychiatric reports. Empirical studies indicate that courts are increasingly encountering such data, particularly in cases of homicide, severe violence, insanity evaluations, and capital sentencing. Methods/Results: Neuroimaging can document brain disease or injury that may impair cognition, affect regulation, or impulse control, important to assessment of criminal accountability, and future risk. Biological markers may corroborate psychiatric and neurological diagnoses, and as such make expert reasoning more transparent. Assessments that omit biological investigations may be considered incomplete, with a risk of undermining fair judicial process and equality before the law, as well as hampering future treatment and risk management. However, there are notable challenges that limit the use of biomarkers in forensic psychiatry evaluations. Neuroscientific findings have methodological and inferential limitations affecting their practical use in individual cases. Furthermore, there is a mismatch between the empirical nature of neuroscience and the normative legal questions such as those of criminal accountability and intent. Additional concerns include risks of over-interpretation (“brain overclaim”), “double-edged sword” effects (mitigation versus inferred dangerousness), inequitable access, and the absence of robust, jurisdiction-specific standards. Conclusions: The inclusion of biological markers in forensic psychiatry evaluations has considerable potential benefits but also raises challenges that need to be addressed in future practice, research, and guideline development.

]]> Should Biological Markers Be Included in Forensic Psychiatric Reports? Unn Kristin Hauvik doi: 10.3390/proceedings2026150013 Proceedings 2026-09-01 Proceedings 2026-09-01 150 1 Abstract 13 10.3390/proceedings2026150013 https://www.mdpi.com/2504-3900/150/1/13 Proceedings, Vol. 148, Pages 5: Too Human to Resist: AI Anthropomorphism and Manipulation Vulnerability in Gen Z https://www.mdpi.com/2504-3900/148/1/5 Conversational AI has entered adolescents’ everyday routines as something more intimate than a search engine and less accountable than a human adviser. Based on a free association task with Gen Z participants and focus group interviews with high school students, this study examines how young users describe AI when they speak about thinking, trust, emotional safety, and influence. The findings reveal a recurrent tension: AI is valued for speed, availability, and patience while sometimes being associated with hallucination, reduced effort, dependency, and personal exposure. The article explores how perceived manipulation vulnerability may be shaped by two connected pathways: cognitive reliance on unstable knowledge and emotional trust in human-like interaction. The exploratory study draws on a free association task with 125 Gen Z participants and four focus groups with 46 high school students aged 16–18. 2026-08-31 Proceedings, Vol. 148, Pages 5: Too Human to Resist: AI Anthropomorphism and Manipulation Vulnerability in Gen Z

Proceedings doi: 10.3390/proceedings2026148005

Authors: Veronika László Lucia Bubrik Zoltán András Dániel

Conversational AI has entered adolescents’ everyday routines as something more intimate than a search engine and less accountable than a human adviser. Based on a free association task with Gen Z participants and focus group interviews with high school students, this study examines how young users describe AI when they speak about thinking, trust, emotional safety, and influence. The findings reveal a recurrent tension: AI is valued for speed, availability, and patience while sometimes being associated with hallucination, reduced effort, dependency, and personal exposure. The article explores how perceived manipulation vulnerability may be shaped by two connected pathways: cognitive reliance on unstable knowledge and emotional trust in human-like interaction. The exploratory study draws on a free association task with 125 Gen Z participants and four focus groups with 46 high school students aged 16–18.

]]> Too Human to Resist: AI Anthropomorphism and Manipulation Vulnerability in Gen Z Veronika László Lucia Bubrik Zoltán András Dániel doi: 10.3390/proceedings2026148005 Proceedings 2026-08-31 Proceedings 2026-08-31 148 1 Proceeding Paper 5 10.3390/proceedings2026148005 https://www.mdpi.com/2504-3900/148/1/5 Proceedings, Vol. 146, Pages 126: Detection of a New Reproductive Location of Twaite Shad (Alosa fallax) with eDNA in the Ter River, NE Catalonia https://www.mdpi.com/2504-3900/146/1/126 The Twaite shad (Alosa fallax Lacépède, 1800) is an endangered anadromous fish listed as Vulnerable on the IUCN Red List. In northern Catalonia (NE Iberian Peninsula), the species maintains a small reproducing population in the Fluvià River, while in the Ter River only sparse, irregular records exist and the status of the species has remained essentially unknown. This study aimed to assess the status of Twaite shad in the lower Ter River through a combined approach of scientific rod-and-reel fishing and eDNA analysis, with the specific objectives of confirming adult presence during the spawning season and detecting juveniles as evidence of effective reproduction. Water samples for eDNA analysis were collected at multiple sites along the lower Ter, across five sampling campaigns conducted between May and September 2025. Duplicate samples of mean 32 L each were filtered in situ and amplified using the MiFish 12S marker with twelve PCR replicates per site and sequenced using Illumina high-throughput technology at a 40 M read sequencing depth per sample. Complementary scientific fishing was carried out using a rod-and-reel technique with demonstrated effectiveness for Twaite shad, comprising five spring sessions (40 h of effort) and four autumn sessions (80 h of effort) at key habitat sites. The eDNA approach proved markedly more sensitive than conventional fishing. Notably, high-resolution eDNA confirmed the presence of the species below the Ullà weir, demonstrating its capacity to move upstream past the Torroella de Montgrí gauging station, most likely facilitated by elevated discharge events during spring. A confirmed eDNA detection in late August provides strong evidence of juvenile presence and, consequently, of effective reproduction in the Ter River. Conclusions: This study provides the first evidence that Twaite shad reproduce in the Ter River, confirming the existence of a previously undescribed breeding population. The eDNA approach demonstrated substantially greater sensitivity than conventional fishing methods for this cryptic migratory species. Priority actions include the identification of spawning grounds, the extension of eDNA surveys to reaches upstream of Ullà—now feasible given the current breach in the weir—and the genetic characterisation of this population to determine its origin, diversity, and potential introgression. 2026-08-31 Proceedings, Vol. 146, Pages 126: Detection of a New Reproductive Location of Twaite Shad (Alosa fallax) with eDNA in the Ter River, NE Catalonia

Proceedings doi: 10.3390/proceedings2026146126

Authors: Miguel A. Fuentes Quim Pou-Rovira Neus Mairal Laia Nogué Jenny Caner Víctor Osorio Teresa Buchaca Marc Ventura

The Twaite shad (Alosa fallax Lacépède, 1800) is an endangered anadromous fish listed as Vulnerable on the IUCN Red List. In northern Catalonia (NE Iberian Peninsula), the species maintains a small reproducing population in the Fluvià River, while in the Ter River only sparse, irregular records exist and the status of the species has remained essentially unknown. This study aimed to assess the status of Twaite shad in the lower Ter River through a combined approach of scientific rod-and-reel fishing and eDNA analysis, with the specific objectives of confirming adult presence during the spawning season and detecting juveniles as evidence of effective reproduction. Water samples for eDNA analysis were collected at multiple sites along the lower Ter, across five sampling campaigns conducted between May and September 2025. Duplicate samples of mean 32 L each were filtered in situ and amplified using the MiFish 12S marker with twelve PCR replicates per site and sequenced using Illumina high-throughput technology at a 40 M read sequencing depth per sample. Complementary scientific fishing was carried out using a rod-and-reel technique with demonstrated effectiveness for Twaite shad, comprising five spring sessions (40 h of effort) and four autumn sessions (80 h of effort) at key habitat sites. The eDNA approach proved markedly more sensitive than conventional fishing. Notably, high-resolution eDNA confirmed the presence of the species below the Ullà weir, demonstrating its capacity to move upstream past the Torroella de Montgrí gauging station, most likely facilitated by elevated discharge events during spring. A confirmed eDNA detection in late August provides strong evidence of juvenile presence and, consequently, of effective reproduction in the Ter River. Conclusions: This study provides the first evidence that Twaite shad reproduce in the Ter River, confirming the existence of a previously undescribed breeding population. The eDNA approach demonstrated substantially greater sensitivity than conventional fishing methods for this cryptic migratory species. Priority actions include the identification of spawning grounds, the extension of eDNA surveys to reaches upstream of Ullà—now feasible given the current breach in the weir—and the genetic characterisation of this population to determine its origin, diversity, and potential introgression.

]]> Detection of a New Reproductive Location of Twaite Shad (Alosa fallax) with eDNA in the Ter River, NE Catalonia Miguel A. Fuentes Quim Pou-Rovira Neus Mairal Laia Nogué Jenny Caner Víctor Osorio Teresa Buchaca Marc Ventura doi: 10.3390/proceedings2026146126 Proceedings 2026-08-31 Proceedings 2026-08-31 146 1 Abstract 126 10.3390/proceedings2026146126 https://www.mdpi.com/2504-3900/146/1/126 Proceedings, Vol. 145, Pages 8: Caregiver-Reported Language Skill Attainment in Arabic-Speaking Saudi Children with Cochlear Implants: A Cross-Sectional Study https://www.mdpi.com/2504-3900/145/1/8 Language outcomes after paediatric cochlear implantation remain under-described in Arabic speaking contexts. This cross-sectional descriptive survey examined caregiver-reported language skill items in a convenience sample of 78 Arabic-speaking Saudi children with cochlear implants, aged 1–5 years. Respondents included parents, other family caregivers, two speech–language specialists, and one teacher; they completed a 28-item Arabic questionnaire developed from prior literature and refined through expert and caregiver review. Responses were scored from never (1) to always (5). Item means ranged from 2.79 to 4.68. Comparatively higher ratings were reported for distinguishing personal belongings and using eye contact, facial expressions, and body movements during play. Comparatively lower ratings occurred for several complex receptive–expressive items, including producing three- to four-word sentences, understanding age-referenced wh-questions, producing 5–10 words after implantation, answering yes/no questions, and using sounds purposefully. In exploratory, unadjusted associations based on self-reported timing categories, higher total scores were associated with longer reported cochlear implant use and younger reported implantation age category. These findings identify items with comparatively lower caregiver ratings in this sample and may inform priorities for future age-stratified research and instrument development in Saudi clinical and educational contexts. 2026-08-28 Proceedings, Vol. 145, Pages 8: Caregiver-Reported Language Skill Attainment in Arabic-Speaking Saudi Children with Cochlear Implants: A Cross-Sectional Study

Proceedings doi: 10.3390/proceedings2026145008

Authors: Taghreed Waslalah Alhzmi

Language outcomes after paediatric cochlear implantation remain under-described in Arabic speaking contexts. This cross-sectional descriptive survey examined caregiver-reported language skill items in a convenience sample of 78 Arabic-speaking Saudi children with cochlear implants, aged 1–5 years. Respondents included parents, other family caregivers, two speech–language specialists, and one teacher; they completed a 28-item Arabic questionnaire developed from prior literature and refined through expert and caregiver review. Responses were scored from never (1) to always (5). Item means ranged from 2.79 to 4.68. Comparatively higher ratings were reported for distinguishing personal belongings and using eye contact, facial expressions, and body movements during play. Comparatively lower ratings occurred for several complex receptive–expressive items, including producing three- to four-word sentences, understanding age-referenced wh-questions, producing 5–10 words after implantation, answering yes/no questions, and using sounds purposefully. In exploratory, unadjusted associations based on self-reported timing categories, higher total scores were associated with longer reported cochlear implant use and younger reported implantation age category. These findings identify items with comparatively lower caregiver ratings in this sample and may inform priorities for future age-stratified research and instrument development in Saudi clinical and educational contexts.

]]> Caregiver-Reported Language Skill Attainment in Arabic-Speaking Saudi Children with Cochlear Implants: A Cross-Sectional Study Taghreed Waslalah Alhzmi doi: 10.3390/proceedings2026145008 Proceedings 2026-08-28 Proceedings 2026-08-28 145 1 Proceeding Paper 8 10.3390/proceedings2026145008 https://www.mdpi.com/2504-3900/145/1/8 Proceedings, Vol. 145, Pages 7: GenAI and Epistemic Practices in Science Education: Opportunities and Didactic Challenges https://www.mdpi.com/2504-3900/145/1/7 Generative artificial intelligence (GenAI) offers opportunities for dialogic scaffolding, scientific reasoning, formative feedback, and instructional design, while raising cognitive, epistemic, didactic, and ethical risks. This review synthesized 34 publications from 2023 to 2026 that were identified through searches of Scopus, Web of Science, and ERIC. Guided by didactic mediation, epistemic practices, and metacognitive regulation, the synthesis indicates that GenAI’s educational contributions depend on three dimensions: learner regulation, teacher mediation, and disciplinary grounding. Six interrelated conditions are organized within these three dimensions to position GenAI as a mediating artefact rather than an epistemic authority. Given the heterogeneous and emerging evidence base, this framework constitutes an interpretive synthesis requiring empirical validation across science education contexts. 2026-08-28 Proceedings, Vol. 145, Pages 7: GenAI and Epistemic Practices in Science Education: Opportunities and Didactic Challenges

Proceedings doi: 10.3390/proceedings2026145007

Authors: Meryem Janati Idrissi Omar Aarab El Mabrouk Nabil Amri

Generative artificial intelligence (GenAI) offers opportunities for dialogic scaffolding, scientific reasoning, formative feedback, and instructional design, while raising cognitive, epistemic, didactic, and ethical risks. This review synthesized 34 publications from 2023 to 2026 that were identified through searches of Scopus, Web of Science, and ERIC. Guided by didactic mediation, epistemic practices, and metacognitive regulation, the synthesis indicates that GenAI’s educational contributions depend on three dimensions: learner regulation, teacher mediation, and disciplinary grounding. Six interrelated conditions are organized within these three dimensions to position GenAI as a mediating artefact rather than an epistemic authority. Given the heterogeneous and emerging evidence base, this framework constitutes an interpretive synthesis requiring empirical validation across science education contexts.

]]> GenAI and Epistemic Practices in Science Education: Opportunities and Didactic Challenges Meryem Janati Idrissi Omar Aarab El Mabrouk Nabil Amri doi: 10.3390/proceedings2026145007 Proceedings 2026-08-28 Proceedings 2026-08-28 145 1 Proceeding Paper 7 10.3390/proceedings2026145007 https://www.mdpi.com/2504-3900/145/1/7 Proceedings, Vol. 139, Pages 30: Statement of Peer Review https://www.mdpi.com/2504-3900/139/1/30 In submitting conference proceedings to Proceedings, the Volume Editors of the proceedings would like to certify to the publisher that all papers published in this volume have been subjected to peer review by the designated expert referees and were administered by the Volume Editors strictly following the policies announced on the conference website [...] 2026-08-28 Proceedings, Vol. 139, Pages 30: Statement of Peer Review

Proceedings doi: 10.3390/proceedings2026139030

Authors: Valentina Berardinetti Pierpaolo Limone Giusi Antonia Toto

In submitting conference proceedings to Proceedings, the Volume Editors of the proceedings would like to certify to the publisher that all papers published in this volume have been subjected to peer review by the designated expert referees and were administered by the Volume Editors strictly following the policies announced on the conference website [...]

]]> Statement of Peer Review Valentina Berardinetti Pierpaolo Limone Giusi Antonia Toto doi: 10.3390/proceedings2026139030 Proceedings 2026-08-28 Proceedings 2026-08-28 139 1 Editorial 30 10.3390/proceedings2026139030 https://www.mdpi.com/2504-3900/139/1/30 Proceedings, Vol. 139, Pages 29: Preface: The Image as a Form of Knowledge: Transdisciplinary Perspectives on Visual Education https://www.mdpi.com/2504-3900/139/1/29 In the contemporary landscape of educational and communication research, characterized by an increasingly visual, interconnected, and constantly evolving ecosystem, Visual Education emerges as an epistemological and operational field of transdisciplinary nature, able to integrate knowledge from pedagogy, digital technologies, visual arts, and cognitive sciences, with the aim of critically interpreting the cultural, cognitive, and social dynamics of contemporaneity [...] 2026-08-28 Proceedings, Vol. 139, Pages 29: Preface: The Image as a Form of Knowledge: Transdisciplinary Perspectives on Visual Education

Proceedings doi: 10.3390/proceedings2026139029

Authors: Valentina Berardinetti Pierpaolo Limone Giusi Antonia Toto

In the contemporary landscape of educational and communication research, characterized by an increasingly visual, interconnected, and constantly evolving ecosystem, Visual Education emerges as an epistemological and operational field of transdisciplinary nature, able to integrate knowledge from pedagogy, digital technologies, visual arts, and cognitive sciences, with the aim of critically interpreting the cultural, cognitive, and social dynamics of contemporaneity [...]

]]> Preface: The Image as a Form of Knowledge: Transdisciplinary Perspectives on Visual Education Valentina Berardinetti Pierpaolo Limone Giusi Antonia Toto doi: 10.3390/proceedings2026139029 Proceedings 2026-08-28 Proceedings 2026-08-28 139 1 Editorial 29 10.3390/proceedings2026139029 https://www.mdpi.com/2504-3900/139/1/29 Proceedings, Vol. 145, Pages 6: Integrating Artificial Intelligence into Irregular Warfare Education: A Framework for Military Learning and Training https://www.mdpi.com/2504-3900/145/1/6 Artificial Intelligence (AI) is increasingly embedded in contemporary military operations, yet its effective and ethical integration depends largely on how defense institutions educate and prepare personnel. While technological advancements receive significant attention, the pedagogical systems that enable responsible adoption remain underexamined. This study examines how AI can be systematically incorporated into professional military education and training for irregular warfare. Using qualitative doctrinal analysis and thematic synthesis of United States and NATO policy documents and scholarly literature, the research develops a five-layer instructional framework linking strategic alignment, doctrinal integration, operational learning, data-driven intelligence education, and human-AI governance. Findings indicate that successful AI adoption requires coordinated reforms in doctrine, instructional design, and ethical oversight rather than isolated technical training, supported by experiential learning and continuous professional development. By reframing AI integration as a pedagogical transformation rather than a purely technical upgrade, this research offers practical guidance for educators, policymakers, and training institutions seeking to enhance operational readiness while preserving transparency, accountability, and principled human judgment. The framework is intended to safeguard against the potential misuse of AI by embedding rigorous ethical governance, accountability, and appropriate human oversight throughout military education and training. 2026-08-27 Proceedings, Vol. 145, Pages 6: Integrating Artificial Intelligence into Irregular Warfare Education: A Framework for Military Learning and Training

Proceedings doi: 10.3390/proceedings2026145006

Authors: Maurice Dawson Samson Quaye

Artificial Intelligence (AI) is increasingly embedded in contemporary military operations, yet its effective and ethical integration depends largely on how defense institutions educate and prepare personnel. While technological advancements receive significant attention, the pedagogical systems that enable responsible adoption remain underexamined. This study examines how AI can be systematically incorporated into professional military education and training for irregular warfare. Using qualitative doctrinal analysis and thematic synthesis of United States and NATO policy documents and scholarly literature, the research develops a five-layer instructional framework linking strategic alignment, doctrinal integration, operational learning, data-driven intelligence education, and human-AI governance. Findings indicate that successful AI adoption requires coordinated reforms in doctrine, instructional design, and ethical oversight rather than isolated technical training, supported by experiential learning and continuous professional development. By reframing AI integration as a pedagogical transformation rather than a purely technical upgrade, this research offers practical guidance for educators, policymakers, and training institutions seeking to enhance operational readiness while preserving transparency, accountability, and principled human judgment. The framework is intended to safeguard against the potential misuse of AI by embedding rigorous ethical governance, accountability, and appropriate human oversight throughout military education and training.

]]> Integrating Artificial Intelligence into Irregular Warfare Education: A Framework for Military Learning and Training Maurice Dawson Samson Quaye doi: 10.3390/proceedings2026145006 Proceedings 2026-08-27 Proceedings 2026-08-27 145 1 Proceeding Paper 6 10.3390/proceedings2026145006 https://www.mdpi.com/2504-3900/145/1/6 Proceedings, Vol. 150, Pages 12: Involving Service Users in Research in Forensic Psychiatry https://www.mdpi.com/2504-3900/150/1/12 Background: The involvement of people with lived experience in psychiatric research is gaining momentum internationally, yet forensic psychiatry remains largely absent from this development—particularly in Germany. Highly regulated settings, stigma, and institutional barriers make meaningful service user involvement both rare and challenging. The PART Advisory Board serves as a best practice model for conducting research not merely about service users, but genuinely with them—as co-researchers throughout the entire research process. Methods: The PART Advisory Board brings together researchers and individuals with lived experience, including current inpatients and formerly detained persons. Involvement spans all phases of research—from topic identification and study design to recruitment and data collection. Examples include structured project presentations with guideline-based feedback aimed at achieving active participation within entire projects—such as serving as an expert panel for guidelines on VR use in forensic settings. Board members also revise patient information materials, co-develop flyers, and present them as co-speakers at conferences. Results: This multi-level involvement improves research quality, relevance, and acceptance. Researchers gain practice-oriented perspectives that refine methods and strengthen interpretation. Board members experience empowerment through recognition of their expertise. Initial analyses confirm that participatory structures are feasible in highly regulated forensic contexts when roles are clear, attitudes are respectful, and communication is continuous. Conclusions: The PART Advisory Board demonstrates that participatory research in forensic psychiatry is both achievable and enriching. Successful implementation requires tailored processes, flexible structures, and individual support throughout the research journey—offering a transferable model for embedding service user involvement as a permanent part of forensic research and practice. 2026-08-27 Proceedings, Vol. 150, Pages 12: Involving Service Users in Research in Forensic Psychiatry

Proceedings doi: 10.3390/proceedings2026150012

Authors: Eva Drewelow Birgit Völlm

Background: The involvement of people with lived experience in psychiatric research is gaining momentum internationally, yet forensic psychiatry remains largely absent from this development—particularly in Germany. Highly regulated settings, stigma, and institutional barriers make meaningful service user involvement both rare and challenging. The PART Advisory Board serves as a best practice model for conducting research not merely about service users, but genuinely with them—as co-researchers throughout the entire research process. Methods: The PART Advisory Board brings together researchers and individuals with lived experience, including current inpatients and formerly detained persons. Involvement spans all phases of research—from topic identification and study design to recruitment and data collection. Examples include structured project presentations with guideline-based feedback aimed at achieving active participation within entire projects—such as serving as an expert panel for guidelines on VR use in forensic settings. Board members also revise patient information materials, co-develop flyers, and present them as co-speakers at conferences. Results: This multi-level involvement improves research quality, relevance, and acceptance. Researchers gain practice-oriented perspectives that refine methods and strengthen interpretation. Board members experience empowerment through recognition of their expertise. Initial analyses confirm that participatory structures are feasible in highly regulated forensic contexts when roles are clear, attitudes are respectful, and communication is continuous. Conclusions: The PART Advisory Board demonstrates that participatory research in forensic psychiatry is both achievable and enriching. Successful implementation requires tailored processes, flexible structures, and individual support throughout the research journey—offering a transferable model for embedding service user involvement as a permanent part of forensic research and practice.

]]> Involving Service Users in Research in Forensic Psychiatry Eva Drewelow Birgit Völlm doi: 10.3390/proceedings2026150012 Proceedings 2026-08-27 Proceedings 2026-08-27 150 1 Abstract 12 10.3390/proceedings2026150012 https://www.mdpi.com/2504-3900/150/1/12 Proceedings, Vol. 150, Pages 11: Multimodal Correlates of Aggression Potential in Early-Stage Psychosis: Integrating Brain Structure, Polygenic Risk, and Social Adversity https://www.mdpi.com/2504-3900/150/1/11 Introduction: Aggression in early-stage mental disorders imposes a substantial clinical and societal burden and is shaped by environmental, neurobiological, and genetic factors. Understanding their joint contribution could aid risk assessment and early intervention. Methods: We used data from the prospective, multicentric European PRONIA project, comprising 402 minimally medicated participants (mean [SD] age = 25.34 [5.84]; female sex = 49.5%) with clinical high-risk states for psychosis (n = 116), recent-onset depression (n = 146) or psychosis (n = 140). Multi-block sparse partial least squares (MB-SPLS) algorithm was employed within a nested cross-validation framework to detect multivariate associations between aggression potential (excited component of the PANSS), social adversity, structural and diffusion MRI, and polygenic risk. Post hoc analyses examined specificity to overall illness severity and associations with personality, emotion recognition, coping, resilience, and functional remission. Results: MB-SPLS linked higher aggression potential in recent-onset psychosis to greater discrimination, emotional and physical trauma, bullying, higher polygenic risk for anxiety, schizophrenia, tension, and bipolar disorder, ventricular enlargement, reduced fronto-parietal-temporal white matter volume, and microstructural alterations of commissural, association, and limbic tracts. This signature was related to, but not fully explained by, illness severity and was associated with lower agreeableness, impaired emotion recognition, and poorer functional remission. Conclusions: Aggression potential in early psychosis corresponds to a distinct, multilevel biotype spanning environmental adversity, genetic liability, and predominantly white matter alterations, and is related to but not reducible to overall illness severity. Associations with social cognition, personality, and functional outcome highlight its clinical relevance and potential target points for early identification and intervention. 2026-08-27 Proceedings, Vol. 150, Pages 11: Multimodal Correlates of Aggression Potential in Early-Stage Psychosis: Integrating Brain Structure, Polygenic Risk, and Social Adversity

Proceedings doi: 10.3390/proceedings2026150011

Authors: Clara Weyer Clara Vetter Jaroslav Rokicki Unn Kristin Haukvik Kolja Schiltz Peter Falkai Nikolaos Koutsouleris David Popovic the PRONIA Consortium the PRONIA Consortium

Introduction: Aggression in early-stage mental disorders imposes a substantial clinical and societal burden and is shaped by environmental, neurobiological, and genetic factors. Understanding their joint contribution could aid risk assessment and early intervention. Methods: We used data from the prospective, multicentric European PRONIA project, comprising 402 minimally medicated participants (mean [SD] age = 25.34 [5.84]; female sex = 49.5%) with clinical high-risk states for psychosis (n = 116), recent-onset depression (n = 146) or psychosis (n = 140). Multi-block sparse partial least squares (MB-SPLS) algorithm was employed within a nested cross-validation framework to detect multivariate associations between aggression potential (excited component of the PANSS), social adversity, structural and diffusion MRI, and polygenic risk. Post hoc analyses examined specificity to overall illness severity and associations with personality, emotion recognition, coping, resilience, and functional remission. Results: MB-SPLS linked higher aggression potential in recent-onset psychosis to greater discrimination, emotional and physical trauma, bullying, higher polygenic risk for anxiety, schizophrenia, tension, and bipolar disorder, ventricular enlargement, reduced fronto-parietal-temporal white matter volume, and microstructural alterations of commissural, association, and limbic tracts. This signature was related to, but not fully explained by, illness severity and was associated with lower agreeableness, impaired emotion recognition, and poorer functional remission. Conclusions: Aggression potential in early psychosis corresponds to a distinct, multilevel biotype spanning environmental adversity, genetic liability, and predominantly white matter alterations, and is related to but not reducible to overall illness severity. Associations with social cognition, personality, and functional outcome highlight its clinical relevance and potential target points for early identification and intervention.

]]> Multimodal Correlates of Aggression Potential in Early-Stage Psychosis: Integrating Brain Structure, Polygenic Risk, and Social Adversity Clara Weyer Clara Vetter Jaroslav Rokicki Unn Kristin Haukvik Kolja Schiltz Peter Falkai Nikolaos Koutsouleris David Popovic the PRONIA Consortium the PRONIA Consortium doi: 10.3390/proceedings2026150011 Proceedings 2026-08-27 Proceedings 2026-08-27 150 1 Abstract 11 10.3390/proceedings2026150011 https://www.mdpi.com/2504-3900/150/1/11 Proceedings, Vol. 150, Pages 10: Gaze Behavior Towards Children in Pedophilia and Hebephilia https://www.mdpi.com/2504-3900/150/1/10 Background: Indirect measurement methods play a central role in the assessment of deviant sexual interests, including pedohebephilia. Among these approaches, eye-tracking paradigms that assess gaze behavior may serve as indirect indicators of deviant vs. non-deviant sexual interest. However, the diagnostic utility of eye-tracking measures for distinguishing pedohebephilia from sexual interest in adults (teleiophilia) remains unclear. Objective: The aim of this study was to examine differences in gaze behavior among individuals with pedohebephilic and teleiophilic sexual interest and to determine the extent to which eye-tracking measures can differentiate pedophilia and hebephilia from teleiophilia. Method: Gaze behavior was assessed using an eye tracker while images of children were presented alongside images of adults. Gaze measures included the time to first fixation, first fixation count and total duration of fixation. Sexual interest was determined through a clinical interview as part of a standard intake procedure of an outpatient prevention program. Results: The sample comprised men with pedophilia, hebephilia and a teleiophilic control group. Men with pedophilic sexual interest showed a significantly higher number of first fixations and a shorter time to first fixation on child stimuli compared with adult stimuli, while the teleiophilic group showed the opposite pattern. Findings for the hebephilic group were inconclusive. Total fixation duration was more evenly distributed between child and adult stimuli in the pedophilic group, whereas the other two groups showed longer fixation durations for adult stimuli [1]. Conclusions: The findings suggest that eye-tracking may provide a useful additional approach for assessing deviant sexual interest. 2026-08-26 Proceedings, Vol. 150, Pages 10: Gaze Behavior Towards Children in Pedophilia and Hebephilia

Proceedings doi: 10.3390/proceedings2026150010

Authors: Puyan Heindl Kolja Schiltz

Background: Indirect measurement methods play a central role in the assessment of deviant sexual interests, including pedohebephilia. Among these approaches, eye-tracking paradigms that assess gaze behavior may serve as indirect indicators of deviant vs. non-deviant sexual interest. However, the diagnostic utility of eye-tracking measures for distinguishing pedohebephilia from sexual interest in adults (teleiophilia) remains unclear. Objective: The aim of this study was to examine differences in gaze behavior among individuals with pedohebephilic and teleiophilic sexual interest and to determine the extent to which eye-tracking measures can differentiate pedophilia and hebephilia from teleiophilia. Method: Gaze behavior was assessed using an eye tracker while images of children were presented alongside images of adults. Gaze measures included the time to first fixation, first fixation count and total duration of fixation. Sexual interest was determined through a clinical interview as part of a standard intake procedure of an outpatient prevention program. Results: The sample comprised men with pedophilia, hebephilia and a teleiophilic control group. Men with pedophilic sexual interest showed a significantly higher number of first fixations and a shorter time to first fixation on child stimuli compared with adult stimuli, while the teleiophilic group showed the opposite pattern. Findings for the hebephilic group were inconclusive. Total fixation duration was more evenly distributed between child and adult stimuli in the pedophilic group, whereas the other two groups showed longer fixation durations for adult stimuli [1]. Conclusions: The findings suggest that eye-tracking may provide a useful additional approach for assessing deviant sexual interest.

]]> Gaze Behavior Towards Children in Pedophilia and Hebephilia Puyan Heindl Kolja Schiltz doi: 10.3390/proceedings2026150010 Proceedings 2026-08-26 Proceedings 2026-08-26 150 1 Abstract 10 10.3390/proceedings2026150010 https://www.mdpi.com/2504-3900/150/1/10 Proceedings, Vol. 148, Pages 4: A Reinforcement Learning Approach for Climate Change Adaptation and Policy: Using a Deep Q-Network for Multi-Sectorial Resilience in Kitui County, Kenya https://www.mdpi.com/2504-3900/148/1/4 Climate change presents serious challenges, especially in dry and semi-dry areas, and these challenges are most severe in sub-Saharan Africa, where people are dependent on rain-fed agriculture. To address this issue, a new type of reinforcement learning (RL) mechanism that can be used to simulate and optimize climate adaptation policies in Kitui County, Kenya, was developed. In this study, combined geospatial data encompassing several variables, such as precipitation, the health of vegetation, population, and economic indicators, were used to create a custom reinforcement learning environment. A Deep Q-Network (DQN) agent was developed to distribute restricted financial resources amongst nine climate adaptation policies over a 25-year simulation period. The findings clearly show that the artificial intelligence agent targeted the long-term sustainability of the ecosystem. In connection to this, it made the choice of agroforestry in 76.0% of its decision cycles, and at the same time, it tactically bypassed high-capital, high-latency infrastructure such as dams and opted for distributed adaptive measures. This framework provides a powerful tool to support policymakers in a data-driven manner to analyze the interdependence of urgent aid and sustainable development. The study demonstrates the possibilities of using AI-powered decision support systems in the field of climate policy planning for the areas most affected by climate change. 2026-08-24 Proceedings, Vol. 148, Pages 4: A Reinforcement Learning Approach for Climate Change Adaptation and Policy: Using a Deep Q-Network for Multi-Sectorial Resilience in Kitui County, Kenya

Proceedings doi: 10.3390/proceedings2026148004

Authors: Ndinde Eya Ekolle

Climate change presents serious challenges, especially in dry and semi-dry areas, and these challenges are most severe in sub-Saharan Africa, where people are dependent on rain-fed agriculture. To address this issue, a new type of reinforcement learning (RL) mechanism that can be used to simulate and optimize climate adaptation policies in Kitui County, Kenya, was developed. In this study, combined geospatial data encompassing several variables, such as precipitation, the health of vegetation, population, and economic indicators, were used to create a custom reinforcement learning environment. A Deep Q-Network (DQN) agent was developed to distribute restricted financial resources amongst nine climate adaptation policies over a 25-year simulation period. The findings clearly show that the artificial intelligence agent targeted the long-term sustainability of the ecosystem. In connection to this, it made the choice of agroforestry in 76.0% of its decision cycles, and at the same time, it tactically bypassed high-capital, high-latency infrastructure such as dams and opted for distributed adaptive measures. This framework provides a powerful tool to support policymakers in a data-driven manner to analyze the interdependence of urgent aid and sustainable development. The study demonstrates the possibilities of using AI-powered decision support systems in the field of climate policy planning for the areas most affected by climate change.

]]> A Reinforcement Learning Approach for Climate Change Adaptation and Policy: Using a Deep Q-Network for Multi-Sectorial Resilience in Kitui County, Kenya Ndinde Eya Ekolle doi: 10.3390/proceedings2026148004 Proceedings 2026-08-24 Proceedings 2026-08-24 148 1 Proceeding Paper 4 10.3390/proceedings2026148004 https://www.mdpi.com/2504-3900/148/1/4 Proceedings, Vol. 150, Pages 9: Genes and Environment in Shaping Human Behavior: Legal and Forensic Perspectives https://www.mdpi.com/2504-3900/150/1/9 Background: Research in behavioral genetics has demonstrated that genetic factors significantly contribute to individual differences in behavior, while environmental exposures shape gene expression through epigenetic mechanisms. This interaction is also relevant to the development of antisocial behavior and psychopathic traits. One of the first evidence of gene–environment interaction was the association between low-activity variants of the MAOA gene, childhood maltreatment, and increased risk of antisocial behavior [1]. Similarly, our research in incarcerated populations showed that adverse paternal parenting is associated with higher levels of psychopathy and the HTR1B rs13212041 TT genotype appears to modulate the individual susceptibility to negative experiences [2]. Single genetic variants, however, exert only modest effects and current evidence supports a polygenic model in which multiple genetic factors interact with environmental adversity to influence neurodevelopment and behavioral outcomes. Using a genome-wide/endophenotype informed analysis, for example, we identified novel gene–environment interactions as risk factors for psychopathy, involving three independent genetic loci in interaction with paternal maltreatment, which were previously associated with disruptive behavior, temperament, and neuroticism [3]. More recently, we also evaluated whether machine-learning models, integrating behavioral, environmental, and genetic variables, could be helpful to predict psychopathic traits. Methods: We compared logistic regression, random forest, support vector machine, XGBoost, and multilayer perceptron. Results: Support vector machine showed the highest accuracy for predicting Psychopathy Check List-Revised (PCL-R) Factor 2 (antisocial lifestyle). Feature-importance analyses identified impulsivity (BIS-11), empathy (IRI), childhood maltreatment (MOPS), and 12 SNPs as the most informative predictors. Notably, removing genetic variables or MOPS scores substantially reduced the model accuracy, indicating that both genetic and environmental information meaningfully contributed to prediction of antisocial behavior. Conclusions: These findings confirm that genetic influences are neither deterministic nor sufficient to explain criminal behavior but may contribute to interindividual differences in vulnerability, particularly through their interaction with environmental and psychosocial factors. In forensic psychiatry, the integration of genetic and environmental information into behavioral assessment may provide additional objective correlates that complement, rather than replace, traditional clinical and psychosocial evaluations. Such an integrated approach could potentially contribute to a more comprehensive understanding of individual vulnerability and behavioral trajectories. However, the use of genetic information in assessments of criminal responsibility should be approached with caution and proven expertise, given the complex, multifactorial nature of antisocial and criminal behavior. 2026-08-21 Proceedings, Vol. 150, Pages 9: Genes and Environment in Shaping Human Behavior: Legal and Forensic Perspectives

Proceedings doi: 10.3390/proceedings2026150009

Authors: Silvia Pellegrini Sara Palumbo Lucia Billeci

Background: Research in behavioral genetics has demonstrated that genetic factors significantly contribute to individual differences in behavior, while environmental exposures shape gene expression through epigenetic mechanisms. This interaction is also relevant to the development of antisocial behavior and psychopathic traits. One of the first evidence of gene–environment interaction was the association between low-activity variants of the MAOA gene, childhood maltreatment, and increased risk of antisocial behavior [1]. Similarly, our research in incarcerated populations showed that adverse paternal parenting is associated with higher levels of psychopathy and the HTR1B rs13212041 TT genotype appears to modulate the individual susceptibility to negative experiences [2]. Single genetic variants, however, exert only modest effects and current evidence supports a polygenic model in which multiple genetic factors interact with environmental adversity to influence neurodevelopment and behavioral outcomes. Using a genome-wide/endophenotype informed analysis, for example, we identified novel gene–environment interactions as risk factors for psychopathy, involving three independent genetic loci in interaction with paternal maltreatment, which were previously associated with disruptive behavior, temperament, and neuroticism [3]. More recently, we also evaluated whether machine-learning models, integrating behavioral, environmental, and genetic variables, could be helpful to predict psychopathic traits. Methods: We compared logistic regression, random forest, support vector machine, XGBoost, and multilayer perceptron. Results: Support vector machine showed the highest accuracy for predicting Psychopathy Check List-Revised (PCL-R) Factor 2 (antisocial lifestyle). Feature-importance analyses identified impulsivity (BIS-11), empathy (IRI), childhood maltreatment (MOPS), and 12 SNPs as the most informative predictors. Notably, removing genetic variables or MOPS scores substantially reduced the model accuracy, indicating that both genetic and environmental information meaningfully contributed to prediction of antisocial behavior. Conclusions: These findings confirm that genetic influences are neither deterministic nor sufficient to explain criminal behavior but may contribute to interindividual differences in vulnerability, particularly through their interaction with environmental and psychosocial factors. In forensic psychiatry, the integration of genetic and environmental information into behavioral assessment may provide additional objective correlates that complement, rather than replace, traditional clinical and psychosocial evaluations. Such an integrated approach could potentially contribute to a more comprehensive understanding of individual vulnerability and behavioral trajectories. However, the use of genetic information in assessments of criminal responsibility should be approached with caution and proven expertise, given the complex, multifactorial nature of antisocial and criminal behavior.

]]> Genes and Environment in Shaping Human Behavior: Legal and Forensic Perspectives Silvia Pellegrini Sara Palumbo Lucia Billeci doi: 10.3390/proceedings2026150009 Proceedings 2026-08-21 Proceedings 2026-08-21 150 1 Abstract 9 10.3390/proceedings2026150009 https://www.mdpi.com/2504-3900/150/1/9 Proceedings, Vol. 150, Pages 8: Predictors of Violence in Schizophrenia Spectrum Disorders: A Multimodal Approach https://www.mdpi.com/2504-3900/150/1/8 Background: The risk of violence is elevated in patients with Schizophrenia Spectrum Disorders (SSD). However, current violence risk assessment approaches rely predominantly on clinical and historical factors and remain limited in predictive accuracy, highlighting the need for more objective complementary markers. Methods: The present study examined the neurobiological differences between violent (n = 66) and non-violent (n = 166) SSD patients, and compared the predictive utility of these neurobiological markers and clinical risk factors, including childhood trauma and psychopathology, in distinguishing violent from non-violent individuals. We further aimed to construct a multimodal predictive model using the strongest predictors from both domains, evaluating whether combining neural and clinical variables improves predictive performance beyond either single domain. Structural and resting-state MRI scans were acquired for patients and healthy controls (n = 504), who were used to calibrate the normative model. Neuroimaging data were analyzed within a normative modelling framework, and univariate associations between neurobiological and clinical factors and violence were examined. Results: A multimodal model integrating neurobiological and clinical measures achieved moderate classification performance (74.7% balanced accuracy), outperforming all individual predictors. The strongest predictors included the medial orbitofrontal cortex, caudal middle frontal gyrus, limbic-default mode network functional connectivity, and childhood sexual abuse. Conclusions: Our findings suggest that alterations in fronto-limbic systems involved in goal-directed decision-making and emotion regulation may contribute to violent behaviour in SSD. Furthermore, the improved performance of the multimodal model supports the potential added utility of integrating neuroimaging markers with established clinical risk factors in violence risk assessment. 2026-08-20 Proceedings, Vol. 150, Pages 8: Predictors of Violence in Schizophrenia Spectrum Disorders: A Multimodal Approach

Proceedings doi: 10.3390/proceedings2026150008

Authors: Aline Huynh Unn K. Haukvik Megan Campbell Kristien van der Walt Jaroslav Rokicki

Background: The risk of violence is elevated in patients with Schizophrenia Spectrum Disorders (SSD). However, current violence risk assessment approaches rely predominantly on clinical and historical factors and remain limited in predictive accuracy, highlighting the need for more objective complementary markers. Methods: The present study examined the neurobiological differences between violent (n = 66) and non-violent (n = 166) SSD patients, and compared the predictive utility of these neurobiological markers and clinical risk factors, including childhood trauma and psychopathology, in distinguishing violent from non-violent individuals. We further aimed to construct a multimodal predictive model using the strongest predictors from both domains, evaluating whether combining neural and clinical variables improves predictive performance beyond either single domain. Structural and resting-state MRI scans were acquired for patients and healthy controls (n = 504), who were used to calibrate the normative model. Neuroimaging data were analyzed within a normative modelling framework, and univariate associations between neurobiological and clinical factors and violence were examined. Results: A multimodal model integrating neurobiological and clinical measures achieved moderate classification performance (74.7% balanced accuracy), outperforming all individual predictors. The strongest predictors included the medial orbitofrontal cortex, caudal middle frontal gyrus, limbic-default mode network functional connectivity, and childhood sexual abuse. Conclusions: Our findings suggest that alterations in fronto-limbic systems involved in goal-directed decision-making and emotion regulation may contribute to violent behaviour in SSD. Furthermore, the improved performance of the multimodal model supports the potential added utility of integrating neuroimaging markers with established clinical risk factors in violence risk assessment.

]]> Predictors of Violence in Schizophrenia Spectrum Disorders: A Multimodal Approach Aline Huynh Unn K. Haukvik Megan Campbell Kristien van der Walt Jaroslav Rokicki doi: 10.3390/proceedings2026150008 Proceedings 2026-08-20 Proceedings 2026-08-20 150 1 Abstract 8 10.3390/proceedings2026150008 https://www.mdpi.com/2504-3900/150/1/8 Proceedings, Vol. 150, Pages 4: Patterns of Substance Use in a Forensic Psychiatric Hospital in Germany https://www.mdpi.com/2504-3900/150/1/4 Background: This presentation is based on a paper that was published last year in the peer-reviewed journal Recht & Psychiatrie [1]. The study investigates consumption patterns in patients who have been diagnosed with a substance use disorder and are being treated in a forensic psychiatric hospital under section 64 of the German penal code. High cost of treatment and a higher risk of reoffending when terminated prematurely underlines the importance of determining who has a high chance of completing treatment and why. It is important to note, that the investigation of dynamic predictors of recidivism in patients who undergo mandatory forensic addiction treatment has been insufficiently examined. One dynamic factor could be the consumption pattern during treatment. Relapses during addiction treatment is not uncommon and relapse rates range from 16.7 to 81.6%. However, evidence shows that moderate consumption does not necessarily have a negative effect on legal prognosis. So far, there are no clear findings as to how substance use during therapy affects recidivism or early treatment termination. Furthermore, consumption patterns have not yet been conclusively conceptualized. Studies don’t go beyond quantifying relapses and only investigate factors that promote or protect patients from relapsing. So far, there has been no investigation of actual consumption patterns during forensic mandatory treatment. Therefore, the goal of this study was to provide a description and classification of consumption patterns, identify factors that promote substance use or encourage abstinence and describe the relationship between consumption patterns and treatment termination. Methods: This qualitative study examined the components of consumption patterns via 17 interviews with patients, who underwent mandatory treatment in a forensic psychiatric hospital in Germany and two focus groups with employees of the same hospital. Results: Findings show different patterns: temporal, observed and internal. A temporal pattern can be divided into five overarching types (progressive, regressive, constant, intermittent, chaotic). Internal patterns of factors within and outside the patient constitute the observed consumption patterns. Conclusions: The reflection of the observed behavior allows therapeutic intervention points. Future studies should validate the components and further investigate the relationship between certain patterns of consumption behavior and recidivism. 2026-08-19 Proceedings, Vol. 150, Pages 4: Patterns of Substance Use in a Forensic Psychiatric Hospital in Germany

Proceedings doi: 10.3390/proceedings2026150004

Authors: Finn Sörensen Birgit Völlm

Background: This presentation is based on a paper that was published last year in the peer-reviewed journal Recht & Psychiatrie [1]. The study investigates consumption patterns in patients who have been diagnosed with a substance use disorder and are being treated in a forensic psychiatric hospital under section 64 of the German penal code. High cost of treatment and a higher risk of reoffending when terminated prematurely underlines the importance of determining who has a high chance of completing treatment and why. It is important to note, that the investigation of dynamic predictors of recidivism in patients who undergo mandatory forensic addiction treatment has been insufficiently examined. One dynamic factor could be the consumption pattern during treatment. Relapses during addiction treatment is not uncommon and relapse rates range from 16.7 to 81.6%. However, evidence shows that moderate consumption does not necessarily have a negative effect on legal prognosis. So far, there are no clear findings as to how substance use during therapy affects recidivism or early treatment termination. Furthermore, consumption patterns have not yet been conclusively conceptualized. Studies don’t go beyond quantifying relapses and only investigate factors that promote or protect patients from relapsing. So far, there has been no investigation of actual consumption patterns during forensic mandatory treatment. Therefore, the goal of this study was to provide a description and classification of consumption patterns, identify factors that promote substance use or encourage abstinence and describe the relationship between consumption patterns and treatment termination. Methods: This qualitative study examined the components of consumption patterns via 17 interviews with patients, who underwent mandatory treatment in a forensic psychiatric hospital in Germany and two focus groups with employees of the same hospital. Results: Findings show different patterns: temporal, observed and internal. A temporal pattern can be divided into five overarching types (progressive, regressive, constant, intermittent, chaotic). Internal patterns of factors within and outside the patient constitute the observed consumption patterns. Conclusions: The reflection of the observed behavior allows therapeutic intervention points. Future studies should validate the components and further investigate the relationship between certain patterns of consumption behavior and recidivism.

]]> Patterns of Substance Use in a Forensic Psychiatric Hospital in Germany Finn Sörensen Birgit Völlm doi: 10.3390/proceedings2026150004 Proceedings 2026-08-19 Proceedings 2026-08-19 150 1 Abstract 4 10.3390/proceedings2026150004 https://www.mdpi.com/2504-3900/150/1/4 Proceedings, Vol. 150, Pages 6: Immunological and Lipid Metabolic Markers of Aggression https://www.mdpi.com/2504-3900/150/1/6 Background: Aggression and violence are complex behavioral phenomena with substantial biological underpinnings involving immune activation and lipid metabolism. Previous evidence has linked inflammatory signaling and altered cholesterol homeostasis to impulsive and antisocial behaviors. The present exploratory study investigated differential gene expression patterns associated with severe violent offending and their relationship with aggression-related phenotypes. Methods: Whole-blood samples from 19 male severe violent offenders and 11 healthy controls were analyzed using Clariom S gene expression arrays followed by qPCR validation. Participants additionally completed psychometric assessments including the State Trait Anger Expression Inventory-II (STAXI-II), the Psychopathic Personality Inventory-Revised (PPI-R), and the Appetitive and Facilitative Aggression Scale (AFAS). Results: Differential expression analysis identified 833 significantly dysregulated genes (FDR < 0.1; fold change ≥ 1.5) in violent offenders relative to controls. Among the most strongly altered genes were IL1B, TLR4, PTGS2 (COX-2), ABCA1, IL6R, PRF1, and MMP9. qPCR validation confirmed significant upregulation of MMP9, PTGS2, IL6R, and PRF1. Gene x behavior analyses demonstrated significant associations between PTGS2 expression and reactive as well as outwardly directed aggression, while MMP9 expression correlated with psychopathy-related traits. Furthermore, several lipid metabolism-related genes, including ABCA1, ABCG1, NPC1, DGAT2, and TLR4, showed expression patterns suggestive of altered intracellular lipid trafficking and lysosomal lipid accumulation. ABCA1 expression correlated positively with outward anger expression (STAXI-II), whereas ABCG1 and TLR4 were negatively associated with inwardly expressed anger (STAXI-II). Conclusions: These findings support a model in which innate immune activation and dysregulated lipid metabolism contribute to aggression-related phenotypes and severe violent behavior. Although exploratory and limited by sample size, the results highlight potential molecular pathways underlying pathological aggression and warrant further investigation in larger cohorts. 2026-08-19 Proceedings, Vol. 150, Pages 6: Immunological and Lipid Metabolic Markers of Aggression

Proceedings doi: 10.3390/proceedings2026150006

Authors: Michael Fritz Karlheinz Holzmann Priscila Batista da Rosa Judith Streb David Engblom Manuela Dudeck

Background: Aggression and violence are complex behavioral phenomena with substantial biological underpinnings involving immune activation and lipid metabolism. Previous evidence has linked inflammatory signaling and altered cholesterol homeostasis to impulsive and antisocial behaviors. The present exploratory study investigated differential gene expression patterns associated with severe violent offending and their relationship with aggression-related phenotypes. Methods: Whole-blood samples from 19 male severe violent offenders and 11 healthy controls were analyzed using Clariom S gene expression arrays followed by qPCR validation. Participants additionally completed psychometric assessments including the State Trait Anger Expression Inventory-II (STAXI-II), the Psychopathic Personality Inventory-Revised (PPI-R), and the Appetitive and Facilitative Aggression Scale (AFAS). Results: Differential expression analysis identified 833 significantly dysregulated genes (FDR < 0.1; fold change ≥ 1.5) in violent offenders relative to controls. Among the most strongly altered genes were IL1B, TLR4, PTGS2 (COX-2), ABCA1, IL6R, PRF1, and MMP9. qPCR validation confirmed significant upregulation of MMP9, PTGS2, IL6R, and PRF1. Gene x behavior analyses demonstrated significant associations between PTGS2 expression and reactive as well as outwardly directed aggression, while MMP9 expression correlated with psychopathy-related traits. Furthermore, several lipid metabolism-related genes, including ABCA1, ABCG1, NPC1, DGAT2, and TLR4, showed expression patterns suggestive of altered intracellular lipid trafficking and lysosomal lipid accumulation. ABCA1 expression correlated positively with outward anger expression (STAXI-II), whereas ABCG1 and TLR4 were negatively associated with inwardly expressed anger (STAXI-II). Conclusions: These findings support a model in which innate immune activation and dysregulated lipid metabolism contribute to aggression-related phenotypes and severe violent behavior. Although exploratory and limited by sample size, the results highlight potential molecular pathways underlying pathological aggression and warrant further investigation in larger cohorts.

]]> Immunological and Lipid Metabolic Markers of Aggression Michael Fritz Karlheinz Holzmann Priscila Batista da Rosa Judith Streb David Engblom Manuela Dudeck doi: 10.3390/proceedings2026150006 Proceedings 2026-08-19 Proceedings 2026-08-19 150 1 Abstract 6 10.3390/proceedings2026150006 https://www.mdpi.com/2504-3900/150/1/6 Proceedings, Vol. 150, Pages 7: Long Lasting Impacts: The Role of Childhood Adversity for Treatment Completion in Prison Rehabilitation https://www.mdpi.com/2504-3900/150/1/7 Background: People in prison have a higher prevalence of adverse childhood experiences (ACEs) compared to the general population. This has been found across different cultural contexts. While treatment programs for people in prison have moderate completion rates, the impact of ACEs on peoples’ treatment and outcomes has been neglected in research, especially for men. Methods/Results: In a series of studies with men attending violent offending treatment programs, it was shown that the ability to complete treatment was substantially impacted by adverse childhood experiences. The association between diver-sity of childhood adversities and treatment outcomes was not better explained by posttraumatic symptoms or antisocial, borderline or psychopathic personality traits. Conclusions: Replication of these associations in various cultural contexts, implications for forensic assessment and treatment practice should be considered. Future research should investigate the pathways for these associations, including physiological and hormonal changes due to the adverse experiences, and acute stress reactivity in interpersonal interactions. 2026-08-19 Proceedings, Vol. 150, Pages 7: Long Lasting Impacts: The Role of Childhood Adversity for Treatment Completion in Prison Rehabilitation

Proceedings doi: 10.3390/proceedings2026150007

Authors: Hedwig Eisenbarth

Background: People in prison have a higher prevalence of adverse childhood experiences (ACEs) compared to the general population. This has been found across different cultural contexts. While treatment programs for people in prison have moderate completion rates, the impact of ACEs on peoples’ treatment and outcomes has been neglected in research, especially for men. Methods/Results: In a series of studies with men attending violent offending treatment programs, it was shown that the ability to complete treatment was substantially impacted by adverse childhood experiences. The association between diver-sity of childhood adversities and treatment outcomes was not better explained by posttraumatic symptoms or antisocial, borderline or psychopathic personality traits. Conclusions: Replication of these associations in various cultural contexts, implications for forensic assessment and treatment practice should be considered. Future research should investigate the pathways for these associations, including physiological and hormonal changes due to the adverse experiences, and acute stress reactivity in interpersonal interactions.

]]> Long Lasting Impacts: The Role of Childhood Adversity for Treatment Completion in Prison Rehabilitation Hedwig Eisenbarth doi: 10.3390/proceedings2026150007 Proceedings 2026-08-19 Proceedings 2026-08-19 150 1 Abstract 7 10.3390/proceedings2026150007 https://www.mdpi.com/2504-3900/150/1/7 Proceedings, Vol. 150, Pages 5: Ethical Pros and Cons of Research on Particularly Vulnerable Patient Populations https://www.mdpi.com/2504-3900/150/1/5 Background: Research involving particularly vulnerable patient populations represents a central ethical challenge in contemporary medicine. On the one hand, inclusion of these groups is essential to ensure evidence-based care and to prevent health inequities. On the other hand, they require special protection due to limited autonomy, structural dependency, and increased risk of exploitation. Methods: This presentation examines the tension between necessary inclusion and ethical risk through the lens of core bioethical principles and current research ethics frameworks. Forensic psychiatry serves as a paradigmatic example, as clinical care, research, and legal control are closely intertwined. Results: The presentation focuses on three key dimensions of risk: limited autonomy, potential exploitation, and distorted consent processes. It further highlights that ethical risks are not uniform but vary depending on the type of research, such as therapeutic interventions, neurobiological studies, or risk assessment research. This underscores the need for nuanced ethical evaluation rather than categorical inclusion or exclusion. Conclusions: The presentation argues that the ethical imperative is not to exclude vulnerable populations from research, but to include them under conditions that ensure transparency, fairness, and heightened protection. Practical approaches to ethical responsible research include enhanced consent procedures, independent advocacy, and context-sensitive study design. 2026-08-19 Proceedings, Vol. 150, Pages 5: Ethical Pros and Cons of Research on Particularly Vulnerable Patient Populations

Proceedings doi: 10.3390/proceedings2026150005

Authors: Irina Franke

Background: Research involving particularly vulnerable patient populations represents a central ethical challenge in contemporary medicine. On the one hand, inclusion of these groups is essential to ensure evidence-based care and to prevent health inequities. On the other hand, they require special protection due to limited autonomy, structural dependency, and increased risk of exploitation. Methods: This presentation examines the tension between necessary inclusion and ethical risk through the lens of core bioethical principles and current research ethics frameworks. Forensic psychiatry serves as a paradigmatic example, as clinical care, research, and legal control are closely intertwined. Results: The presentation focuses on three key dimensions of risk: limited autonomy, potential exploitation, and distorted consent processes. It further highlights that ethical risks are not uniform but vary depending on the type of research, such as therapeutic interventions, neurobiological studies, or risk assessment research. This underscores the need for nuanced ethical evaluation rather than categorical inclusion or exclusion. Conclusions: The presentation argues that the ethical imperative is not to exclude vulnerable populations from research, but to include them under conditions that ensure transparency, fairness, and heightened protection. Practical approaches to ethical responsible research include enhanced consent procedures, independent advocacy, and context-sensitive study design.

]]> Ethical Pros and Cons of Research on Particularly Vulnerable Patient Populations Irina Franke doi: 10.3390/proceedings2026150005 Proceedings 2026-08-19 Proceedings 2026-08-19 150 1 Abstract 5 10.3390/proceedings2026150005 https://www.mdpi.com/2504-3900/150/1/5 Proceedings, Vol. 145, Pages 5: Developer Education for Sustainable Web Development: Evidence from the Balkan Region https://www.mdpi.com/2504-3900/145/1/5 Sustainable web development focuses on designing web pages that minimize their environmental impact by reducing digital carbon emissions while maintaining optimal website performance and user experience. This study emphasizes the necessity of adopting sustainable web development practices in the Balkan region, highlighting the role of institutional education and web developers in encouraging environmentally responsible web practices within a Technology-Enhanced Education context. A descriptive cross-country analysis was conducted on a total of 8976 websites across five Balkan countries—Albania, Greece, Serbia, Montenegro, and North Macedonia—selected from public domain registries and categorized by sector (education, organization, government, and commercial domains). Data were collected during December 2025. DNS verification was conducted to identify active domains, and hosting sustainability was determined through the Green Web Foundation API. “Green hosting” is operationally defined as hosting that is completely powered by renewable energy sources. Overall, the findings suggest that the prevalence of green hosting in the region is very low, at approximately 27.77%, with substantial variation across sectors. Educational institutions (.edu) and non-profit organizations (.org) have higher adoption rates, while public administrations (.gov) and small commercial websites generally fall behind. The country with the highest percentage of green hosting is Greece (64.07%), while the lowest is Albania, with less than 1%. These findings reveal significant gaps in knowledge and practices among developers and institutions, indicating an urgent need for targeted educational interventions. Crucially, this study connects empirical evidence directly to education. Recognized knowledge gaps guide curriculum development and the design of practical training modules to equip developers and institutional personnel with the skills to adopt sustainable web practices. This research establishes a regional comparative baseline and offers practical recommendations for incorporating sustainability into web development education. The findings endorse strategies and policies aimed at promoting environmentally sustainable digital infrastructure in the Balkan area. 2026-08-19 Proceedings, Vol. 145, Pages 5: Developer Education for Sustainable Web Development: Evidence from the Balkan Region

Proceedings doi: 10.3390/proceedings2026145005

Authors: Alma Sheko

Sustainable web development focuses on designing web pages that minimize their environmental impact by reducing digital carbon emissions while maintaining optimal website performance and user experience. This study emphasizes the necessity of adopting sustainable web development practices in the Balkan region, highlighting the role of institutional education and web developers in encouraging environmentally responsible web practices within a Technology-Enhanced Education context. A descriptive cross-country analysis was conducted on a total of 8976 websites across five Balkan countries—Albania, Greece, Serbia, Montenegro, and North Macedonia—selected from public domain registries and categorized by sector (education, organization, government, and commercial domains). Data were collected during December 2025. DNS verification was conducted to identify active domains, and hosting sustainability was determined through the Green Web Foundation API. “Green hosting” is operationally defined as hosting that is completely powered by renewable energy sources. Overall, the findings suggest that the prevalence of green hosting in the region is very low, at approximately 27.77%, with substantial variation across sectors. Educational institutions (.edu) and non-profit organizations (.org) have higher adoption rates, while public administrations (.gov) and small commercial websites generally fall behind. The country with the highest percentage of green hosting is Greece (64.07%), while the lowest is Albania, with less than 1%. These findings reveal significant gaps in knowledge and practices among developers and institutions, indicating an urgent need for targeted educational interventions. Crucially, this study connects empirical evidence directly to education. Recognized knowledge gaps guide curriculum development and the design of practical training modules to equip developers and institutional personnel with the skills to adopt sustainable web practices. This research establishes a regional comparative baseline and offers practical recommendations for incorporating sustainability into web development education. The findings endorse strategies and policies aimed at promoting environmentally sustainable digital infrastructure in the Balkan area.

]]> Developer Education for Sustainable Web Development: Evidence from the Balkan Region Alma Sheko doi: 10.3390/proceedings2026145005 Proceedings 2026-08-19 Proceedings 2026-08-19 145 1 Proceeding Paper 5 10.3390/proceedings2026145005 https://www.mdpi.com/2504-3900/145/1/5 Proceedings, Vol. 150, Pages 3: A Latent Class Analysis of Mental Disorders, Substance Use, and Aggressive Antisocial Behaviour Among Swedish Forensic Psychiatric Patients https://www.mdpi.com/2504-3900/150/1/3 Patients in forensic mental health services (FMHS) with co-occurring mental disorders (MD), substance use disorders (SUD), and a history of violent crime—often referred to as the “triply troubled”—represent one of the most clinically complex and poorly researched patient populations. Standard diagnostics tend to treat this group as homogeneous, which may impede the development of patient-centred treatment and risk management. This study aimed to identify patient subgroups within this population based on patterns of MDs, substance use history, and aggressive antisocial behaviour (AAB). A latent class analysis (LCA) was conducted using clinical data from 119 patients at a high-security FMHS facility in Sweden. Indicator variables included lifetime diagnoses of SUD and MDs and types of substances used. History of AAB and criminal offending were used as distal outcomes. A four-class solution provided the best model fit. Class 1 (polysubstance, high AAB; 100% SUD probability) and Class 4 (polysubstance, bipolar disorder; 49% SUD probability) exhibited pronounced criminal and AAB history. Class 2 (psychosis and cannabis-limited substance use) and Class 3 (autism and limited substance use) represented less criminologically ladened subgroups. Notably, Classes 1 and 4 shared extensive AAB histories despite differing SUD diagnostic probabilities, highlighting the limitations of diagnosis alone as a basis for risk assessment. The four-class typology offers a clinically meaningful framework for stratifying triply troubled patients in FMHS, aligning with the Risk–Need–Responsivity model and severity-graded treatment recommendations The typology has direct implications for treatment differentiation, relapse prevention, and risk management in forensic psychiatric settings. 2026-08-17 Proceedings, Vol. 150, Pages 3: A Latent Class Analysis of Mental Disorders, Substance Use, and Aggressive Antisocial Behaviour Among Swedish Forensic Psychiatric Patients

Proceedings doi: 10.3390/proceedings2026150003

Authors: Johan Green Ann-Sophie Lindqvist Bagge Natalie Laporte Peter Andiné Märta Wallinius Malin Hildebrand Karlén

Patients in forensic mental health services (FMHS) with co-occurring mental disorders (MD), substance use disorders (SUD), and a history of violent crime—often referred to as the “triply troubled”—represent one of the most clinically complex and poorly researched patient populations. Standard diagnostics tend to treat this group as homogeneous, which may impede the development of patient-centred treatment and risk management. This study aimed to identify patient subgroups within this population based on patterns of MDs, substance use history, and aggressive antisocial behaviour (AAB). A latent class analysis (LCA) was conducted using clinical data from 119 patients at a high-security FMHS facility in Sweden. Indicator variables included lifetime diagnoses of SUD and MDs and types of substances used. History of AAB and criminal offending were used as distal outcomes. A four-class solution provided the best model fit. Class 1 (polysubstance, high AAB; 100% SUD probability) and Class 4 (polysubstance, bipolar disorder; 49% SUD probability) exhibited pronounced criminal and AAB history. Class 2 (psychosis and cannabis-limited substance use) and Class 3 (autism and limited substance use) represented less criminologically ladened subgroups. Notably, Classes 1 and 4 shared extensive AAB histories despite differing SUD diagnostic probabilities, highlighting the limitations of diagnosis alone as a basis for risk assessment. The four-class typology offers a clinically meaningful framework for stratifying triply troubled patients in FMHS, aligning with the Risk–Need–Responsivity model and severity-graded treatment recommendations The typology has direct implications for treatment differentiation, relapse prevention, and risk management in forensic psychiatric settings.

]]> A Latent Class Analysis of Mental Disorders, Substance Use, and Aggressive Antisocial Behaviour Among Swedish Forensic Psychiatric Patients Johan Green Ann-Sophie Lindqvist Bagge Natalie Laporte Peter Andiné Märta Wallinius Malin Hildebrand Karlén doi: 10.3390/proceedings2026150003 Proceedings 2026-08-17 Proceedings 2026-08-17 150 1 Abstract 3 10.3390/proceedings2026150003 https://www.mdpi.com/2504-3900/150/1/3 Proceedings, Vol. 140, Pages 10: Statement of Peer Review https://www.mdpi.com/2504-3900/140/1/10 In submitting conference proceedings to Proceedings, the volume editors of the proceedings certify to the publisher that all papers published in this volume have been subjected to peer review administered by the volume editors [...] 2026-08-17 Proceedings, Vol. 140, Pages 10: Statement of Peer Review

Proceedings doi: 10.3390/proceedings2026140010

Authors: Isabel María García Sánchez

In submitting conference proceedings to Proceedings, the volume editors of the proceedings certify to the publisher that all papers published in this volume have been subjected to peer review administered by the volume editors [...]

]]> Statement of Peer Review Isabel María García Sánchez doi: 10.3390/proceedings2026140010 Proceedings 2026-08-17 Proceedings 2026-08-17 140 1 Editorial 10 10.3390/proceedings2026140010 https://www.mdpi.com/2504-3900/140/1/10 Proceedings, Vol. 140, Pages 9: Preface of the 1st International Online Conference on Administrative Sciences (IOCAS 2026) https://www.mdpi.com/2504-3900/140/1/9 The 1st International Online Conference on Administrative Sciences (IOCAS 2026) was hosted online from 4 to 5 February 2026 [...] 2026-08-17 Proceedings, Vol. 140, Pages 9: Preface of the 1st International Online Conference on Administrative Sciences (IOCAS 2026)

Proceedings doi: 10.3390/proceedings2026140009

Authors: Isabel María García Sánchez

The 1st International Online Conference on Administrative Sciences (IOCAS 2026) was hosted online from 4 to 5 February 2026 [...]

]]> Preface of the 1st International Online Conference on Administrative Sciences (IOCAS 2026) Isabel María García Sánchez doi: 10.3390/proceedings2026140009 Proceedings 2026-08-17 Proceedings 2026-08-17 140 1 Editorial 9 10.3390/proceedings2026140009 https://www.mdpi.com/2504-3900/140/1/9 Proceedings, Vol. 150, Pages 2: Developing a National Database on Forensic-Psychiatric Patients: The CONNECT Study https://www.mdpi.com/2504-3900/150/1/2 Background: Forensic concepts should be guided by current evidence. However, there is currently a lack of even basic information on individuals who have committed a criminal offence in relation to a mental disorder in Germany. Methods: The CONNECT project aims to add knowledge on this patient group by conducting an annual survey of all forensic psychiatric hospitals in Germany. The resulting database will include comprehensive clinical, legal and socio-demographic data. The project aims are (a) to offer a much-needed overview of the sociodemographic, clinical, offending and treatment data of this group, (b) to identify predicting factors associated with leave status and discharge, and (c) to establish rates of reconviction and its predictors at a three-year follow-up. Results: This contribution examines the procedures and obstacles associated with the establishment of the survey. These include concerns about data protection, the necessity of engaging and persuading the ministries of each federal German state, and the scarcity of resources within hospitals, as well as hospital organization and communication. The objective is to foster an understanding of the implementation of a national reference date survey. Conclusions: It is anticipated that this project will have a substantial impact within clinical and academic contexts, while also offering value to policymakers and care providers. 2026-08-17 Proceedings, Vol. 150, Pages 2: Developing a National Database on Forensic-Psychiatric Patients: The CONNECT Study

Proceedings doi: 10.3390/proceedings2026150002

Authors: Katja Köppen Birgit Völlm

Background: Forensic concepts should be guided by current evidence. However, there is currently a lack of even basic information on individuals who have committed a criminal offence in relation to a mental disorder in Germany. Methods: The CONNECT project aims to add knowledge on this patient group by conducting an annual survey of all forensic psychiatric hospitals in Germany. The resulting database will include comprehensive clinical, legal and socio-demographic data. The project aims are (a) to offer a much-needed overview of the sociodemographic, clinical, offending and treatment data of this group, (b) to identify predicting factors associated with leave status and discharge, and (c) to establish rates of reconviction and its predictors at a three-year follow-up. Results: This contribution examines the procedures and obstacles associated with the establishment of the survey. These include concerns about data protection, the necessity of engaging and persuading the ministries of each federal German state, and the scarcity of resources within hospitals, as well as hospital organization and communication. The objective is to foster an understanding of the implementation of a national reference date survey. Conclusions: It is anticipated that this project will have a substantial impact within clinical and academic contexts, while also offering value to policymakers and care providers.

]]> Developing a National Database on Forensic-Psychiatric Patients: The CONNECT Study Katja Köppen Birgit Völlm doi: 10.3390/proceedings2026150002 Proceedings 2026-08-17 Proceedings 2026-08-17 150 1 Abstract 2 10.3390/proceedings2026150002 https://www.mdpi.com/2504-3900/150/1/2 Proceedings, Vol. 148, Pages 3: Navigating the Paradox of Progress: A Qualitative Study of Women’s Entrepreneurship in Morocco https://www.mdpi.com/2504-3900/148/1/3 This study examines how Moroccan women reconcile entrepreneurial endeavors with domestic and social responsibilities. Adopting a qualitative relational framework, the research draws on semi-structured interviews conducted with 17 female entrepreneurs across three regions. The findings indicate that entrepreneurship is positively associated with expanding women’s economic participation and enhancing their social status. However, the persistence of entrenched societal skepticism, disproportionate domestic responsibilities and exclusionary male-centric business networks constitutes a structural barrier to both policy designs and outcomes. The study concludes that women’s entrepreneurship in Morocco represents a social activism that requires a restructuring of intersectional realities to challenge the gendered status quo. 2026-08-14 Proceedings, Vol. 148, Pages 3: Navigating the Paradox of Progress: A Qualitative Study of Women’s Entrepreneurship in Morocco

Proceedings doi: 10.3390/proceedings2026148003

Authors: Najoua Khayati Abdelali Lahrech

This study examines how Moroccan women reconcile entrepreneurial endeavors with domestic and social responsibilities. Adopting a qualitative relational framework, the research draws on semi-structured interviews conducted with 17 female entrepreneurs across three regions. The findings indicate that entrepreneurship is positively associated with expanding women’s economic participation and enhancing their social status. However, the persistence of entrenched societal skepticism, disproportionate domestic responsibilities and exclusionary male-centric business networks constitutes a structural barrier to both policy designs and outcomes. The study concludes that women’s entrepreneurship in Morocco represents a social activism that requires a restructuring of intersectional realities to challenge the gendered status quo.

]]> Navigating the Paradox of Progress: A Qualitative Study of Women’s Entrepreneurship in Morocco Najoua Khayati Abdelali Lahrech doi: 10.3390/proceedings2026148003 Proceedings 2026-08-14 Proceedings 2026-08-14 148 1 Proceeding Paper 3 10.3390/proceedings2026148003 https://www.mdpi.com/2504-3900/148/1/3 Proceedings, Vol. 150, Pages 1: Preface of the Global Minds in Forensic Psychiatry—Advancing Research and Practice (GMFP 2026) https://www.mdpi.com/2504-3900/150/1/1 The field of forensic psychiatry holds a unique position at the intersection of mental health care, neuroscience, psychology, criminology, ethics, and law [...] 2026-08-13 Proceedings, Vol. 150, Pages 1: Preface of the Global Minds in Forensic Psychiatry—Advancing Research and Practice (GMFP 2026)

Proceedings doi: 10.3390/proceedings2026150001

Authors: Michael Fritz Christian Götzl Manuela Dudeck

The field of forensic psychiatry holds a unique position at the intersection of mental health care, neuroscience, psychology, criminology, ethics, and law [...]

]]> Preface of the Global Minds in Forensic Psychiatry—Advancing Research and Practice (GMFP 2026) Michael Fritz Christian Götzl Manuela Dudeck doi: 10.3390/proceedings2026150001 Proceedings 2026-08-13 Proceedings 2026-08-13 150 1 Editorial 1 10.3390/proceedings2026150001 https://www.mdpi.com/2504-3900/150/1/1 Proceedings, Vol. 145, Pages 4: Transforming Initial Wonder into Structured Understanding: A Descriptive Report of an Experiential Science Outreach Station https://www.mdpi.com/2504-3900/145/1/4 The transition from secondary school to science, technology, engineering, and mathematics (STEM) higher education may be hindered when science is perceived as abstract or disconnected from everyday experience. This descriptive educational-practice report presents Beyond the Magic, a participatory science station implemented during the annual ConCiencia FACTA outreach event at the Faculty of Food Science and Technology, National University of Comahue, Argentina. Approximately 200 students aged 15–18 from eight regional secondary schools attended the event in October 2025. During approximately 45-min visits in groups of 10–15, students encountered eight short chemistry and physics experiences organized around prediction, observation, guided explanation, and discussion. Retrospective facilitator observations were organized into descriptive categories: predictions and alternative explanations, spontaneous questions, everyday-life connections, changes in willingness to participate, questions about university study and research, and participation by accompanying teachers. The facilitators perceived broader participation as each visit progressed. These observations were not counted, coded, independently assessed, or collected as contemporaneous transcripts, and no measures of learning, situational interest, STEM identity, or educational aspirations were administered. The contribution is therefore a replicable implementation design rather than evidence of educational effectiveness or changes in STEM pathways. 2026-08-12 Proceedings, Vol. 145, Pages 4: Transforming Initial Wonder into Structured Understanding: A Descriptive Report of an Experiential Science Outreach Station

Proceedings doi: 10.3390/proceedings2026145004

Authors: Facundo Carlos Namor Carolina Antonella Paulino

The transition from secondary school to science, technology, engineering, and mathematics (STEM) higher education may be hindered when science is perceived as abstract or disconnected from everyday experience. This descriptive educational-practice report presents Beyond the Magic, a participatory science station implemented during the annual ConCiencia FACTA outreach event at the Faculty of Food Science and Technology, National University of Comahue, Argentina. Approximately 200 students aged 15–18 from eight regional secondary schools attended the event in October 2025. During approximately 45-min visits in groups of 10–15, students encountered eight short chemistry and physics experiences organized around prediction, observation, guided explanation, and discussion. Retrospective facilitator observations were organized into descriptive categories: predictions and alternative explanations, spontaneous questions, everyday-life connections, changes in willingness to participate, questions about university study and research, and participation by accompanying teachers. The facilitators perceived broader participation as each visit progressed. These observations were not counted, coded, independently assessed, or collected as contemporaneous transcripts, and no measures of learning, situational interest, STEM identity, or educational aspirations were administered. The contribution is therefore a replicable implementation design rather than evidence of educational effectiveness or changes in STEM pathways.

]]> Transforming Initial Wonder into Structured Understanding: A Descriptive Report of an Experiential Science Outreach Station Facundo Carlos Namor Carolina Antonella Paulino doi: 10.3390/proceedings2026145004 Proceedings 2026-08-12 Proceedings 2026-08-12 145 1 Proceeding Paper 4 10.3390/proceedings2026145004 https://www.mdpi.com/2504-3900/145/1/4 Proceedings, Vol. 146, Pages 125: Nocturnal Hypoxia Reduces Aerobic Scope and Telencephalon Size in Cleaner Wrasse Despite Enhanced Inhibitory Control https://www.mdpi.com/2504-3900/146/1/125 Ocean deoxygenation is intensifying globally, and hypoxia is increasingly documented on tropical coral reefs; yet, its consequences for the physiology and social functioning of reef fishes remain poorly understood. Here, we experimentally exposed the bluestreak cleaner wrasse (Labroides dimidiatus) to two repeated nocturnal severe hypoxia events (<2 mg O2 L−1), reflecting the upper range of currently observed reef conditions. We quantified aerobic performance using intermittent flow respirometry across three phases: baseline (n = 24, prior to exposure), post-hypoxia (n = 24, the morning following the first hypoxia event), and post-recovery (n = 22, three days later). Control fish were followed over the same time frame and tested with the same protocol to account for handling or timing effects. Cooperative performance (n = 22) was assessed separately using an ecologically relevant Feeding Against Preference (FAP) task conducted after the second hypoxic night, allowing us to test whether inhibitory control is sensitive to acute oxygen limitation. Brain morphology (n = 12) was subsequently quantified in a subset of individuals to assess potential changes in relative brain size and regional investment. Repeated hypoxia was associated with persistent reductions in aerobic performance. Maximum and resting metabolic rates declined across phases in hypoxia-exposed fish, resulting in reduced absolute and factorial aerobic scope relative to controls. Hypoxia also altered performance in the FAP task, with exposed fish feeding more often against their food preference, a result consistent with enhanced inhibitory control. In contrast, hypoxia exposure was associated with a significant reduction in relative telencephalon size, while other brain regions and overall brain size were unaffected. These findings suggest that severe nocturnal hypoxic events can impose lasting constraints on aerobic capacity and selectively alter brain structure, while also modifying self-control during cooperative interactions. Together, these results indicate that oxygen limitation can affect cognition both directly and through underlying physiological and neural mechanisms. As hypoxic events intensify under ongoing climate change, such constraints may have important implications for the stability of cleaner–client mutualisms, which could ultimately affect broader reef-level ecosystem stability. 2026-08-12 Proceedings, Vol. 146, Pages 125: Nocturnal Hypoxia Reduces Aerobic Scope and Telencephalon Size in Cleaner Wrasse Despite Enhanced Inhibitory Control

Proceedings doi: 10.3390/proceedings2026146125

Authors: Beatriz P. Pereira Catarina Barros Rui Rosa José Ricardo Paula

Ocean deoxygenation is intensifying globally, and hypoxia is increasingly documented on tropical coral reefs; yet, its consequences for the physiology and social functioning of reef fishes remain poorly understood. Here, we experimentally exposed the bluestreak cleaner wrasse (Labroides dimidiatus) to two repeated nocturnal severe hypoxia events (<2 mg O2 L−1), reflecting the upper range of currently observed reef conditions. We quantified aerobic performance using intermittent flow respirometry across three phases: baseline (n = 24, prior to exposure), post-hypoxia (n = 24, the morning following the first hypoxia event), and post-recovery (n = 22, three days later). Control fish were followed over the same time frame and tested with the same protocol to account for handling or timing effects. Cooperative performance (n = 22) was assessed separately using an ecologically relevant Feeding Against Preference (FAP) task conducted after the second hypoxic night, allowing us to test whether inhibitory control is sensitive to acute oxygen limitation. Brain morphology (n = 12) was subsequently quantified in a subset of individuals to assess potential changes in relative brain size and regional investment. Repeated hypoxia was associated with persistent reductions in aerobic performance. Maximum and resting metabolic rates declined across phases in hypoxia-exposed fish, resulting in reduced absolute and factorial aerobic scope relative to controls. Hypoxia also altered performance in the FAP task, with exposed fish feeding more often against their food preference, a result consistent with enhanced inhibitory control. In contrast, hypoxia exposure was associated with a significant reduction in relative telencephalon size, while other brain regions and overall brain size were unaffected. These findings suggest that severe nocturnal hypoxic events can impose lasting constraints on aerobic capacity and selectively alter brain structure, while also modifying self-control during cooperative interactions. Together, these results indicate that oxygen limitation can affect cognition both directly and through underlying physiological and neural mechanisms. As hypoxic events intensify under ongoing climate change, such constraints may have important implications for the stability of cleaner–client mutualisms, which could ultimately affect broader reef-level ecosystem stability.

]]> Nocturnal Hypoxia Reduces Aerobic Scope and Telencephalon Size in Cleaner Wrasse Despite Enhanced Inhibitory Control Beatriz P. Pereira Catarina Barros Rui Rosa José Ricardo Paula doi: 10.3390/proceedings2026146125 Proceedings 2026-08-12 Proceedings 2026-08-12 146 1 Abstract 125 10.3390/proceedings2026146125 https://www.mdpi.com/2504-3900/146/1/125 Proceedings, Vol. 141, Pages 4: Adaptive Performance and Cognitive Regulation in Immersive VR Vocational Training https://www.mdpi.com/2504-3900/141/1/4 Immersive virtual reality (VR) provides a valuable platform for examining cognitive regulation and adaptive performance in realistic vocational tasks. This study investigated adaptation in a hand-tracked VR coffee-preparation task across two trials: a fully guided trial and a subsequent reduced-guidance trial. Thirty participants (N = 30) completed both trials. Performance was assessed using NASA-TLX-based subjective workload ratings, confidence, perceived interaction difficulty, segment-level task times, total completion time, and object-drop errors. Results showed significant reductions in workload, interaction difficulty, insertion, grinding, tamping, and total completion times, as well as fewer dropped objects, while confidence increased. Prior VR experience was associated with lower interaction difficulty, whereas coffee-related experience was associated with higher confidence. 2026-08-12 Proceedings, Vol. 141, Pages 4: Adaptive Performance and Cognitive Regulation in Immersive VR Vocational Training

Proceedings doi: 10.3390/proceedings2026141004

Authors: Irene Ioannidou Spiros Sirmakessis Maria Rigou

Immersive virtual reality (VR) provides a valuable platform for examining cognitive regulation and adaptive performance in realistic vocational tasks. This study investigated adaptation in a hand-tracked VR coffee-preparation task across two trials: a fully guided trial and a subsequent reduced-guidance trial. Thirty participants (N = 30) completed both trials. Performance was assessed using NASA-TLX-based subjective workload ratings, confidence, perceived interaction difficulty, segment-level task times, total completion time, and object-drop errors. Results showed significant reductions in workload, interaction difficulty, insertion, grinding, tamping, and total completion times, as well as fewer dropped objects, while confidence increased. Prior VR experience was associated with lower interaction difficulty, whereas coffee-related experience was associated with higher confidence.

]]> Adaptive Performance and Cognitive Regulation in Immersive VR Vocational Training Irene Ioannidou Spiros Sirmakessis Maria Rigou doi: 10.3390/proceedings2026141004 Proceedings 2026-08-12 Proceedings 2026-08-12 141 1 Proceeding Paper 4 10.3390/proceedings2026141004 https://www.mdpi.com/2504-3900/141/1/4 Proceedings, Vol. 145, Pages 3: Enhancing Statistical Learning in Primary Education Through Scratch: An Educational Intervention Study https://www.mdpi.com/2504-3900/145/1/3 The teaching of statistics in primary education remains underdeveloped, despite its relevance today and its importance in developing students’ statistical literacy. Furthermore, difficulties in mathematics are linked to negative attitudes and low motivation, which has driven the use of ICT as a teaching resource. In this context, Scratch emerges as a promising tool for facilitating learning through block-based programming, although its impact on motivation is still debatable. This study analyses a quasi-experimental intervention using Scratch compared to traditional methodology in the fifth grade of primary school, evaluating academic performance, conceptual understanding, attitudes and motivation toward mathematics, and perceived usability. The results show significant improvements in learning and performance, as well as greater perceived usability, but no change in motivation. 2026-08-11 Proceedings, Vol. 145, Pages 3: Enhancing Statistical Learning in Primary Education Through Scratch: An Educational Intervention Study

Proceedings doi: 10.3390/proceedings2026145003

Authors: Francisco Zambrano-Romero Adrián Gordillo-Merino

The teaching of statistics in primary education remains underdeveloped, despite its relevance today and its importance in developing students’ statistical literacy. Furthermore, difficulties in mathematics are linked to negative attitudes and low motivation, which has driven the use of ICT as a teaching resource. In this context, Scratch emerges as a promising tool for facilitating learning through block-based programming, although its impact on motivation is still debatable. This study analyses a quasi-experimental intervention using Scratch compared to traditional methodology in the fifth grade of primary school, evaluating academic performance, conceptual understanding, attitudes and motivation toward mathematics, and perceived usability. The results show significant improvements in learning and performance, as well as greater perceived usability, but no change in motivation.

]]> Enhancing Statistical Learning in Primary Education Through Scratch: An Educational Intervention Study Francisco Zambrano-Romero Adrián Gordillo-Merino doi: 10.3390/proceedings2026145003 Proceedings 2026-08-11 Proceedings 2026-08-11 145 1 Proceeding Paper 3 10.3390/proceedings2026145003 https://www.mdpi.com/2504-3900/145/1/3 Proceedings, Vol. 140, Pages 8: Cognitive Extensions in Entrepreneurship: A Systematic Review of Artificial Intelligence as a Support System for Entrepreneurial Thinking https://www.mdpi.com/2504-3900/140/1/8 As entrepreneurial environments become increasingly data-intensive, this study examines the literature on artificial intelligence in entrepreneurship through the perspective of cognitive extension. Positioned at the intersection of entrepreneurship and intelligent systems, it investigates the thematic structure and intellectual evolution of this research domain through a systematic bibliometric approach based on Scopus data (1990–2026), incorporating descriptive analysis, keyword co-occurrence mapping, and citation-based techniques. The findings suggest that the literature is organized around artificial intelligence, machine learning, and entrepreneurship, with a pronounced emphasis on decision-making in early-stage venture contexts. Highly cited contributions consistently frame artificial intelligence as a decision-support mechanism that enhances predictive capacity and information processing while remaining contingent upon human judgment. The results further indicate a field characterized by rapid expansion, increasing collaboration, and substantial conceptual diversity. Overall, the study interprets these developments through the cognitive extension perspective, suggesting that the literature increasingly conceptualizes artificial intelligence as a complementary resource that supports, rather than substitutes for, entrepreneurial reasoning. 2026-08-10 Proceedings, Vol. 140, Pages 8: Cognitive Extensions in Entrepreneurship: A Systematic Review of Artificial Intelligence as a Support System for Entrepreneurial Thinking

Proceedings doi: 10.3390/proceedings2026140008

Authors: Aristidis Bitzenis Nikos Koutsoupias Marios Nosios

As entrepreneurial environments become increasingly data-intensive, this study examines the literature on artificial intelligence in entrepreneurship through the perspective of cognitive extension. Positioned at the intersection of entrepreneurship and intelligent systems, it investigates the thematic structure and intellectual evolution of this research domain through a systematic bibliometric approach based on Scopus data (1990–2026), incorporating descriptive analysis, keyword co-occurrence mapping, and citation-based techniques. The findings suggest that the literature is organized around artificial intelligence, machine learning, and entrepreneurship, with a pronounced emphasis on decision-making in early-stage venture contexts. Highly cited contributions consistently frame artificial intelligence as a decision-support mechanism that enhances predictive capacity and information processing while remaining contingent upon human judgment. The results further indicate a field characterized by rapid expansion, increasing collaboration, and substantial conceptual diversity. Overall, the study interprets these developments through the cognitive extension perspective, suggesting that the literature increasingly conceptualizes artificial intelligence as a complementary resource that supports, rather than substitutes for, entrepreneurial reasoning.

]]> Cognitive Extensions in Entrepreneurship: A Systematic Review of Artificial Intelligence as a Support System for Entrepreneurial Thinking Aristidis Bitzenis Nikos Koutsoupias Marios Nosios doi: 10.3390/proceedings2026140008 Proceedings 2026-08-10 Proceedings 2026-08-10 140 1 Proceeding Paper 8 10.3390/proceedings2026140008 https://www.mdpi.com/2504-3900/140/1/8 Proceedings, Vol. 145, Pages 2: Metaverse Adoption in Built-Environment Education: A Kirkpatrick Model-Based Evaluation of Educator Training Outcomes https://www.mdpi.com/2504-3900/145/1/2 The rapid digitalisation of the construction and built-environment sectors has increased demand for innovative teaching methods in higher education. Among emerging digital tools, metaverse applications offer immersive and interactive learning environments with the potential to transform traditional pedagogical practices. This study evaluates the adoption of the metaverse application (EON-XR) in built-environment education by examining educator training outcomes using the Kirkpatrick model. The evaluation focuses on a structured training workshop designed to equip built-environment educators with the skills to integrate metaverse-based applications into first-year teaching modules. A post-training evaluation employed a Google Forms survey instrument explicitly aligned with Kirkpatrick’s four evaluation levels: reaction, learning, behaviour, and results, with Levels 3 (Behaviour) and 4 (Results) operationalised as behavioural-intention and perceived-results proxies collected immediately post-training, ahead of any classroom implementation. The instrument captured educators’ perceptions of usability, relevance, pedagogical value, behavioural intention to apply acquired skills, and the broader institutional and disciplinary implications of metaverse adoption. Both qualitative and quantitative data were analyzed to assess the training programme’s effectiveness across these four dimensions. Findings indicate a generally positive reaction to metaverse training, with educators recognizing the relevance and user-friendliness of the EON-XR application. Evidence of learning was demonstrated by increased confidence and perceived competence in applying metaverse tools for teaching. At the behavioural level, participants expressed strong intentions to integrate metaverse applications into their teaching practice. At the results level, the training was perceived as enhancing teaching engagement, stimulating interest in construction digitalisation, and contributing to the institutional standing of higher education providers. The study concludes that metaverse-based educator training can play a critical role in advancing digital pedagogy within built-environment education. By applying the Kirkpatrick model, this research provides a structured, replicable framework for evaluating the adoption of immersive technology in higher education contexts. 2026-08-06 Proceedings, Vol. 145, Pages 2: Metaverse Adoption in Built-Environment Education: A Kirkpatrick Model-Based Evaluation of Educator Training Outcomes

Proceedings doi: 10.3390/proceedings2026145002

Authors: Olusegun Aanuoluwapo Oguntona

The rapid digitalisation of the construction and built-environment sectors has increased demand for innovative teaching methods in higher education. Among emerging digital tools, metaverse applications offer immersive and interactive learning environments with the potential to transform traditional pedagogical practices. This study evaluates the adoption of the metaverse application (EON-XR) in built-environment education by examining educator training outcomes using the Kirkpatrick model. The evaluation focuses on a structured training workshop designed to equip built-environment educators with the skills to integrate metaverse-based applications into first-year teaching modules. A post-training evaluation employed a Google Forms survey instrument explicitly aligned with Kirkpatrick’s four evaluation levels: reaction, learning, behaviour, and results, with Levels 3 (Behaviour) and 4 (Results) operationalised as behavioural-intention and perceived-results proxies collected immediately post-training, ahead of any classroom implementation. The instrument captured educators’ perceptions of usability, relevance, pedagogical value, behavioural intention to apply acquired skills, and the broader institutional and disciplinary implications of metaverse adoption. Both qualitative and quantitative data were analyzed to assess the training programme’s effectiveness across these four dimensions. Findings indicate a generally positive reaction to metaverse training, with educators recognizing the relevance and user-friendliness of the EON-XR application. Evidence of learning was demonstrated by increased confidence and perceived competence in applying metaverse tools for teaching. At the behavioural level, participants expressed strong intentions to integrate metaverse applications into their teaching practice. At the results level, the training was perceived as enhancing teaching engagement, stimulating interest in construction digitalisation, and contributing to the institutional standing of higher education providers. The study concludes that metaverse-based educator training can play a critical role in advancing digital pedagogy within built-environment education. By applying the Kirkpatrick model, this research provides a structured, replicable framework for evaluating the adoption of immersive technology in higher education contexts.

]]> Metaverse Adoption in Built-Environment Education: A Kirkpatrick Model-Based Evaluation of Educator Training Outcomes Olusegun Aanuoluwapo Oguntona doi: 10.3390/proceedings2026145002 Proceedings 2026-08-06 Proceedings 2026-08-06 145 1 Proceeding Paper 2 10.3390/proceedings2026145002 https://www.mdpi.com/2504-3900/145/1/2 Proceedings, Vol. 149, Pages 1: Abstract of the 1st International Online Conference on Risks, 6–7 July 2026 https://www.mdpi.com/2504-3900/149/1/1 The 1st International Online Conference on Risks (IOCR 2026) was held online from 6–7 July 2026. At a time characterised by intensifying market volatility, rapidly evolving regulatory landscapes, and a proliferation of emerging threats—ranging from climate-related disruptions to cyber vulnerabilities—this conference aims to be a pivotal forum for interdisciplinary dialogue. In a world where risks intertwine and impact every aspect of business, society, and the global economy, the need for collaborative and innovative approaches to risk management has never been more pressing. 2026-08-03 Proceedings, Vol. 149, Pages 1: Abstract of the 1st International Online Conference on Risks, 6–7 July 2026

Proceedings doi: 10.3390/proceedings2026149001

Authors: Steven Haberman

The 1st International Online Conference on Risks (IOCR 2026) was held online from 6–7 July 2026. At a time characterised by intensifying market volatility, rapidly evolving regulatory landscapes, and a proliferation of emerging threats—ranging from climate-related disruptions to cyber vulnerabilities—this conference aims to be a pivotal forum for interdisciplinary dialogue. In a world where risks intertwine and impact every aspect of business, society, and the global economy, the need for collaborative and innovative approaches to risk management has never been more pressing.

]]> Abstract of the 1st International Online Conference on Risks, 6–7 July 2026 Steven Haberman doi: 10.3390/proceedings2026149001 Proceedings 2026-08-03 Proceedings 2026-08-03 149 1 Conference Report 1 10.3390/proceedings2026149001 https://www.mdpi.com/2504-3900/149/1/1 Proceedings, Vol. 140, Pages 7: Abstracts of the 1st International Online Conference on Administrative Sciences (IOCAS 2026) https://www.mdpi.com/2504-3900/140/1/7 The 1st International Online Conference on Administrative Sciences (IOCAS 2026) was hosted online from 4 to 5 February 2026. This conference presented the latest studies in public management, leadership, strategic management and entrepreneurship. The goal was to examine, explore and critically engage with issues and advances in these areas. 2026-07-23 Proceedings, Vol. 140, Pages 7: Abstracts of the 1st International Online Conference on Administrative Sciences (IOCAS 2026)

Proceedings doi: 10.3390/proceedings2026140007

Authors: Isabel María García Sánchez

The 1st International Online Conference on Administrative Sciences (IOCAS 2026) was hosted online from 4 to 5 February 2026. This conference presented the latest studies in public management, leadership, strategic management and entrepreneurship. The goal was to examine, explore and critically engage with issues and advances in these areas.

]]> Abstracts of the 1st International Online Conference on Administrative Sciences (IOCAS 2026) Isabel María García Sánchez doi: 10.3390/proceedings2026140007 Proceedings 2026-07-23 Proceedings 2026-07-23 140 1 Conference Report 7 10.3390/proceedings2026140007 https://www.mdpi.com/2504-3900/140/1/7 Proceedings, Vol. 146, Pages 124: A Strategic Acoustic Telemetry Infrastructure for Marine Biodiversity in the Strait of Gibraltar and Gulf of Cádiz: STRAITS https://www.mdpi.com/2504-3900/146/1/124 Introduction: The Strait of Gibraltar, a narrow gateway between the Atlantic Ocean and the Mediterranean Sea, concentrates intense biotic exchange and functions as a global “ecological gate” for marine species. Despite its ecological relevance, sustained acoustic telemetry monitoring has long been hindered by oceanographic complexity, strong currents, intense maritime traffic, and geopolitical constraints. As its Atlantic approach, the Gulf of Cádiz is a highly productive area supporting diverse habitats, key spawning and nursery grounds, and major migration pathways, while underpinning important commercial fisheries. Objective: to deploy a system to enable continuous monitoring of animal movements across this key biogeographic boundary. Methodology: The EU-funded STRAITS project, a multidisciplinary consortium integrating physical oceanography and marine biotelemetry, has deployed an acoustic telemetry curtain across the Strait of Gibraltar, complemented by a coastal acoustic receiver array in the Gulf of Cádiz. Results: Over the past two years, a consistent pattern of frequent acoustic detections has demonstrated the system’s potential. Data reveal spatio-temporal migration patterns of commercially important species such as Atlantic bluefin tuna (Thunnus thynnus) and meagre (Argyrosomus regius), alongside conservation-relevant species like ocean sunfish (Mola mola). Conclusions: These results highlight the Strait of Gibraltar as a biodiversity hotspot and a strategic location for quantifying connectivity, migration timing, and species fluxes, supporting ecosystem-based management and blue policy frameworks. 2026-07-16 Proceedings, Vol. 146, Pages 124: A Strategic Acoustic Telemetry Infrastructure for Marine Biodiversity in the Strait of Gibraltar and Gulf of Cádiz: STRAITS

Proceedings doi: 10.3390/proceedings2026146124

Authors: César Vilas Miguel Cabanellas-Reboredo David Abecasis Juan Jiménez-Rincón Óscar Mansilla Ricardo F. Sanchez-Leal

Introduction: The Strait of Gibraltar, a narrow gateway between the Atlantic Ocean and the Mediterranean Sea, concentrates intense biotic exchange and functions as a global “ecological gate” for marine species. Despite its ecological relevance, sustained acoustic telemetry monitoring has long been hindered by oceanographic complexity, strong currents, intense maritime traffic, and geopolitical constraints. As its Atlantic approach, the Gulf of Cádiz is a highly productive area supporting diverse habitats, key spawning and nursery grounds, and major migration pathways, while underpinning important commercial fisheries. Objective: to deploy a system to enable continuous monitoring of animal movements across this key biogeographic boundary. Methodology: The EU-funded STRAITS project, a multidisciplinary consortium integrating physical oceanography and marine biotelemetry, has deployed an acoustic telemetry curtain across the Strait of Gibraltar, complemented by a coastal acoustic receiver array in the Gulf of Cádiz. Results: Over the past two years, a consistent pattern of frequent acoustic detections has demonstrated the system’s potential. Data reveal spatio-temporal migration patterns of commercially important species such as Atlantic bluefin tuna (Thunnus thynnus) and meagre (Argyrosomus regius), alongside conservation-relevant species like ocean sunfish (Mola mola). Conclusions: These results highlight the Strait of Gibraltar as a biodiversity hotspot and a strategic location for quantifying connectivity, migration timing, and species fluxes, supporting ecosystem-based management and blue policy frameworks.

]]> A Strategic Acoustic Telemetry Infrastructure for Marine Biodiversity in the Strait of Gibraltar and Gulf of Cádiz: STRAITS César Vilas Miguel Cabanellas-Reboredo David Abecasis Juan Jiménez-Rincón Óscar Mansilla Ricardo F. Sanchez-Leal doi: 10.3390/proceedings2026146124 Proceedings 2026-07-16 Proceedings 2026-07-16 146 1 Abstract 124 10.3390/proceedings2026146124 https://www.mdpi.com/2504-3900/146/1/124 Proceedings, Vol. 139, Pages 28: The Role of Visual Media in Didactic Innovation: Insights from Experimental Pedagogy https://www.mdpi.com/2504-3900/139/1/28 This contribution reports on a Visual Education pilot intervention implemented within the TFA program at the University of Bari (2025). Using a single-group research design (N = 216), the study examines the effects of integrating concept maps, infographics, and videos on disciplinary knowledge, the quality of participants’ outputs, and trainees’ perceptions. Preliminary findings suggest that visual tools may support content comprehension and organization while simultaneously highlighting the importance of structured and reflective visual design in teacher education. 2026-07-16 Proceedings, Vol. 139, Pages 28: The Role of Visual Media in Didactic Innovation: Insights from Experimental Pedagogy

Proceedings doi: 10.3390/proceedings2026139028

Authors: Alberto Fornasari

This contribution reports on a Visual Education pilot intervention implemented within the TFA program at the University of Bari (2025). Using a single-group research design (N = 216), the study examines the effects of integrating concept maps, infographics, and videos on disciplinary knowledge, the quality of participants’ outputs, and trainees’ perceptions. Preliminary findings suggest that visual tools may support content comprehension and organization while simultaneously highlighting the importance of structured and reflective visual design in teacher education.

]]> The Role of Visual Media in Didactic Innovation: Insights from Experimental Pedagogy Alberto Fornasari doi: 10.3390/proceedings2026139028 Proceedings 2026-07-16 Proceedings 2026-07-16 139 1 Proceeding Paper 28 10.3390/proceedings2026139028 https://www.mdpi.com/2504-3900/139/1/28 Proceedings, Vol. 144, Pages 3: Abstracts of the 1st International Online Conference on Societies, 21–23 January 2026 https://www.mdpi.com/2504-3900/144/1/3 Prof [...] 2026-07-10 Proceedings, Vol. 144, Pages 3: Abstracts of the 1st International Online Conference on Societies, 21–23 January 2026

Proceedings doi: 10.3390/proceedings2026144003

Authors: Gregor Wolbring

Prof [...]

]]> Abstracts of the 1st International Online Conference on Societies, 21–23 January 2026 Gregor Wolbring doi: 10.3390/proceedings2026144003 Proceedings 2026-07-10 Proceedings 2026-07-10 144 1 Conference Report 3 10.3390/proceedings2026144003 https://www.mdpi.com/2504-3900/144/1/3 Proceedings, Vol. 141, Pages 3: School Opportunity for Developing Human Intelligence Potential—An Inclusive Case Study Model Based on Organizational Needs Analysis https://www.mdpi.com/2504-3900/141/1/3 This study encourages mainstream schools to adopt inclusion processes aligned with their resources and capacity to foster own inclusive culture. Within a pilot study initially framed as a quasi-experimental design, the project dynamics demonstrated that participatory action research (PAR) enhances the experimental case study through stakeholder reflection. By implementing a Quality Deployment methodology centered on beneficiary needs, innovative solutions emerged for both students at risk of academic failure and those with high abilities, driven by the necessity to maximize their cognitive potential. 2026-07-09 Proceedings, Vol. 141, Pages 3: School Opportunity for Developing Human Intelligence Potential—An Inclusive Case Study Model Based on Organizational Needs Analysis

Proceedings doi: 10.3390/proceedings2026141003

Authors: Loredana Adriana I. Patrascoiu

This study encourages mainstream schools to adopt inclusion processes aligned with their resources and capacity to foster own inclusive culture. Within a pilot study initially framed as a quasi-experimental design, the project dynamics demonstrated that participatory action research (PAR) enhances the experimental case study through stakeholder reflection. By implementing a Quality Deployment methodology centered on beneficiary needs, innovative solutions emerged for both students at risk of academic failure and those with high abilities, driven by the necessity to maximize their cognitive potential.

]]> School Opportunity for Developing Human Intelligence Potential—An Inclusive Case Study Model Based on Organizational Needs Analysis Loredana Adriana I. Patrascoiu doi: 10.3390/proceedings2026141003 Proceedings 2026-07-09 Proceedings 2026-07-09 141 1 Proceeding Paper 3 10.3390/proceedings2026141003 https://www.mdpi.com/2504-3900/141/1/3 Proceedings, Vol. 148, Pages 2: Demographic and Epidemiological Transitions and Implications for Family Support in Brunei Darussalam https://www.mdpi.com/2504-3900/148/1/2 Brunei Darussalam has undergone a rapid demographic and epidemiological transition characterized by declining fertility, increased life expectancy, and a shift from infectious diseases to chronic non-communicable conditions. While these transitions are well described at the population level, their implications for family support systems are less explored. This study synthesizes national data to examine how changes in population structure and disease burden are reshaping caregiving and intergenerational support. Findings highlight shrinking family size, increasing dependency, and growing care complexity. Applying the welfare diamond framework, the analysis shows that these transitions are progressively shifting care responsibilities beyond the traditional family towards greater roles for the state, market and community sectors. These findings suggest that Brunei Darussalam is undergoing an emerging care transition that has important implications for long-term care policy. 2026-07-07 Proceedings, Vol. 148, Pages 2: Demographic and Epidemiological Transitions and Implications for Family Support in Brunei Darussalam

Proceedings doi: 10.3390/proceedings2026148002

Authors: Yi Sing Wee Shyh Poh Teo

Brunei Darussalam has undergone a rapid demographic and epidemiological transition characterized by declining fertility, increased life expectancy, and a shift from infectious diseases to chronic non-communicable conditions. While these transitions are well described at the population level, their implications for family support systems are less explored. This study synthesizes national data to examine how changes in population structure and disease burden are reshaping caregiving and intergenerational support. Findings highlight shrinking family size, increasing dependency, and growing care complexity. Applying the welfare diamond framework, the analysis shows that these transitions are progressively shifting care responsibilities beyond the traditional family towards greater roles for the state, market and community sectors. These findings suggest that Brunei Darussalam is undergoing an emerging care transition that has important implications for long-term care policy.

]]> Demographic and Epidemiological Transitions and Implications for Family Support in Brunei Darussalam Yi Sing Wee Shyh Poh Teo doi: 10.3390/proceedings2026148002 Proceedings 2026-07-07 Proceedings 2026-07-07 148 1 Proceeding Paper 2 10.3390/proceedings2026148002 https://www.mdpi.com/2504-3900/148/1/2 Proceedings, Vol. 140, Pages 6: Evaluating the Impact of Pakistan’s “Digital Pakistan” Initiative on Public Service Delivery https://www.mdpi.com/2504-3900/140/1/6 Digital Pakistan is a pivotal move towards bridging the gap between old bureaucratic methods and new ones, offering a technologically advanced, modernized form of administration for the masses. This paper will critically analyze the initiative’s effects on the delivery of social services to the population through a methodical SWOT analysis, highlighting existing shortcomings, such as insufficient infrastructure and the digital divide, that limit access for all. Based on a qualitative study using secondary data (2018–2024), it was found that although the adoption of more than 40 e-governance solutions and the Digital Pakistan Hub has led to an increase in transparency and responsiveness of the administration, there still remain great threats, such as security breaches in the cyber-sphere and a 67 percent gender gap in access to the Internet. The findings emphasize that although digital tools have helped ease bureaucracy, they are not very effective because of unequal digital literacy. This means that the paper advocates inclusive policymaking that focuses on boosting digital equity and infrastructural resilience to achieve sustainable public-sector reforms. The findings offer a practical roadmap for policymakers to bridge the technological vision with citizen-centric service delivery. 2026-07-06 Proceedings, Vol. 140, Pages 6: Evaluating the Impact of Pakistan’s “Digital Pakistan” Initiative on Public Service Delivery

Proceedings doi: 10.3390/proceedings2026140006

Authors: Saba Farah Hadia Safeer Choudhry Mahnoor Javed

Digital Pakistan is a pivotal move towards bridging the gap between old bureaucratic methods and new ones, offering a technologically advanced, modernized form of administration for the masses. This paper will critically analyze the initiative’s effects on the delivery of social services to the population through a methodical SWOT analysis, highlighting existing shortcomings, such as insufficient infrastructure and the digital divide, that limit access for all. Based on a qualitative study using secondary data (2018–2024), it was found that although the adoption of more than 40 e-governance solutions and the Digital Pakistan Hub has led to an increase in transparency and responsiveness of the administration, there still remain great threats, such as security breaches in the cyber-sphere and a 67 percent gender gap in access to the Internet. The findings emphasize that although digital tools have helped ease bureaucracy, they are not very effective because of unequal digital literacy. This means that the paper advocates inclusive policymaking that focuses on boosting digital equity and infrastructural resilience to achieve sustainable public-sector reforms. The findings offer a practical roadmap for policymakers to bridge the technological vision with citizen-centric service delivery.

]]> Evaluating the Impact of Pakistan’s “Digital Pakistan” Initiative on Public Service Delivery Saba Farah Hadia Safeer Choudhry Mahnoor Javed doi: 10.3390/proceedings2026140006 Proceedings 2026-07-06 Proceedings 2026-07-06 140 1 Proceeding Paper 6 10.3390/proceedings2026140006 https://www.mdpi.com/2504-3900/140/1/6 Proceedings, Vol. 148, Pages 1: Relieving Suffering as a Health System Responsibility: Governance and Global Inequities in Palliative Care https://www.mdpi.com/2504-3900/148/1/1 Palliative care evolved through recognition of subjectively experienced suffering, yet its availability depends on how societies recognize, prioritize and act. This paper presents a narrative, conceptually oriented synthesis of interdisciplinary literature from social science, bioethics and global health policy to examine how concepts of consciousness and suffering shape the governance and distribution of palliative care within health systems, particularly between high-income and low- and middle-income countries. These disparities are mediated by resources, as well as regulatory regimes, financing structures, workforce authority, and political prioritization. Relieving suffering is a health system responsibility that requires policy coherence, proportionate regulation and accountability in addressing global inequities. 2026-07-06 Proceedings, Vol. 148, Pages 1: Relieving Suffering as a Health System Responsibility: Governance and Global Inequities in Palliative Care

Proceedings doi: 10.3390/proceedings2026148001

Authors: Sunjida Shahriah Shyh Poh Teo

Palliative care evolved through recognition of subjectively experienced suffering, yet its availability depends on how societies recognize, prioritize and act. This paper presents a narrative, conceptually oriented synthesis of interdisciplinary literature from social science, bioethics and global health policy to examine how concepts of consciousness and suffering shape the governance and distribution of palliative care within health systems, particularly between high-income and low- and middle-income countries. These disparities are mediated by resources, as well as regulatory regimes, financing structures, workforce authority, and political prioritization. Relieving suffering is a health system responsibility that requires policy coherence, proportionate regulation and accountability in addressing global inequities.

]]> Relieving Suffering as a Health System Responsibility: Governance and Global Inequities in Palliative Care Sunjida Shahriah Shyh Poh Teo doi: 10.3390/proceedings2026148001 Proceedings 2026-07-06 Proceedings 2026-07-06 148 1 Proceeding Paper 1 10.3390/proceedings2026148001 https://www.mdpi.com/2504-3900/148/1/1 Proceedings, Vol. 140, Pages 5: Emotional Intelligence Revisited: Leadership Adaptation in Environments Mediated by Intelligent Technologies https://www.mdpi.com/2504-3900/140/1/5 This study examines the role of emotional intelligence in leadership contexts mediated by intelligent technologies, focusing on how socio-emotional processes are shaped within digital communication environments and artificial intelligence-enabled systems. Situated at the intersection of leadership and technological transformation, the study aims to clarify the thematic structure and intellectual development of this emerging research domain. A systematic bibliometric approach is employed, drawing on data from the Scopus database for 2004–2026 and integrating descriptive analysis, keyword co-occurrence mapping, and co-citation techniques. The results suggest a rapidly expanding yet fragmented body of literature structured around three principal trajectories: leadership in virtual and distributed environments, the integration of emotional processes with intelligent systems, and competence-based approaches linked to digital transformation. Across these strands, emotional intelligence is consistently associated with communication, trust, and team effectiveness, while conditions of mediated interaction increasingly shape its role. The findings further reveal a degree of conceptual dispersion alongside the absence of a standardized vocabulary. Overall, the study concludes that emotional intelligence remains central to leadership effectiveness but is reconfigured as a context-dependent capability embedded within sociotechnical systems. 2026-07-02 Proceedings, Vol. 140, Pages 5: Emotional Intelligence Revisited: Leadership Adaptation in Environments Mediated by Intelligent Technologies

Proceedings doi: 10.3390/proceedings2026140005

Authors: Nikos Koutsoupias Marios Nosios

This study examines the role of emotional intelligence in leadership contexts mediated by intelligent technologies, focusing on how socio-emotional processes are shaped within digital communication environments and artificial intelligence-enabled systems. Situated at the intersection of leadership and technological transformation, the study aims to clarify the thematic structure and intellectual development of this emerging research domain. A systematic bibliometric approach is employed, drawing on data from the Scopus database for 2004–2026 and integrating descriptive analysis, keyword co-occurrence mapping, and co-citation techniques. The results suggest a rapidly expanding yet fragmented body of literature structured around three principal trajectories: leadership in virtual and distributed environments, the integration of emotional processes with intelligent systems, and competence-based approaches linked to digital transformation. Across these strands, emotional intelligence is consistently associated with communication, trust, and team effectiveness, while conditions of mediated interaction increasingly shape its role. The findings further reveal a degree of conceptual dispersion alongside the absence of a standardized vocabulary. Overall, the study concludes that emotional intelligence remains central to leadership effectiveness but is reconfigured as a context-dependent capability embedded within sociotechnical systems.

]]> Emotional Intelligence Revisited: Leadership Adaptation in Environments Mediated by Intelligent Technologies Nikos Koutsoupias Marios Nosios doi: 10.3390/proceedings2026140005 Proceedings 2026-07-02 Proceedings 2026-07-02 140 1 Proceeding Paper 5 10.3390/proceedings2026140005 https://www.mdpi.com/2504-3900/140/1/5 Proceedings, Vol. 147, Pages 1: Stigma as a Behavioral Determinant of Healthy Aging: Ageism and Dementia as Illustrative Contexts https://www.mdpi.com/2504-3900/147/1/1 Stigma, including ageism and dementia-related stigma, operates as a behavioral determinant shaping beliefs, expectations and actions across the life course. This narrative conceptual synthesis draws on World Health Organization frameworks and selected literature to examine how stigma influences behavior at individual and professional levels. Ageism reinforces assumptions of decline and dependency, which may be internalized, reducing engagement in health-promoting behaviors. Dementia stigma contributes to delayed help-seeking, underdiagnosis, and therapeutic nihilism, influencing clinical decision-making and access to rehabilitation and psychosocial interventions. Addressing stigma is critical to modifying behavior, improving care engagement, and supporting inclusive, person-centered approaches to healthy aging and dementia. 2026-06-30 Proceedings, Vol. 147, Pages 1: Stigma as a Behavioral Determinant of Healthy Aging: Ageism and Dementia as Illustrative Contexts

Proceedings doi: 10.3390/proceedings2026147001

Authors: Priyasha Parekh Julia Whyte Shyh Poh Teo

Stigma, including ageism and dementia-related stigma, operates as a behavioral determinant shaping beliefs, expectations and actions across the life course. This narrative conceptual synthesis draws on World Health Organization frameworks and selected literature to examine how stigma influences behavior at individual and professional levels. Ageism reinforces assumptions of decline and dependency, which may be internalized, reducing engagement in health-promoting behaviors. Dementia stigma contributes to delayed help-seeking, underdiagnosis, and therapeutic nihilism, influencing clinical decision-making and access to rehabilitation and psychosocial interventions. Addressing stigma is critical to modifying behavior, improving care engagement, and supporting inclusive, person-centered approaches to healthy aging and dementia.

]]> Stigma as a Behavioral Determinant of Healthy Aging: Ageism and Dementia as Illustrative Contexts Priyasha Parekh Julia Whyte Shyh Poh Teo doi: 10.3390/proceedings2026147001 Proceedings 2026-06-30 Proceedings 2026-06-30 147 1 Proceeding Paper 1 10.3390/proceedings2026147001 https://www.mdpi.com/2504-3900/147/1/1 Proceedings, Vol. 141, Pages 2: Rethinking Mediation in Early Childhood Mathematics: A Conceptual Framework Informed by Inclusive Research https://www.mdpi.com/2504-3900/141/1/2 This paper starts from an understanding of intelligence as learning potential that develops through mediated participation in social, material, and cultural practices. The study examines how mediation is conceptualized in early childhood mathematics education through an integrative literature review of 23 studies across mathematics education and related fields. While semiotic mediation provides a strong theoretical foundation for understanding the role of artefacts, signs, and teacher-guided interaction, the analysis highlights embodied, multimodal, environmental, and relational aspects that often remain implicit in research and practice. Informed by inclusive research, the study offers a conceptual framework for making these aspects more visible. The paper contributes to a more fine-grained understanding of mediation, with implications for inclusive approaches to early childhood mathematics education. 2026-06-29 Proceedings, Vol. 141, Pages 2: Rethinking Mediation in Early Childhood Mathematics: A Conceptual Framework Informed by Inclusive Research

Proceedings doi: 10.3390/proceedings2026141002

Authors: Francesca Granone

This paper starts from an understanding of intelligence as learning potential that develops through mediated participation in social, material, and cultural practices. The study examines how mediation is conceptualized in early childhood mathematics education through an integrative literature review of 23 studies across mathematics education and related fields. While semiotic mediation provides a strong theoretical foundation for understanding the role of artefacts, signs, and teacher-guided interaction, the analysis highlights embodied, multimodal, environmental, and relational aspects that often remain implicit in research and practice. Informed by inclusive research, the study offers a conceptual framework for making these aspects more visible. The paper contributes to a more fine-grained understanding of mediation, with implications for inclusive approaches to early childhood mathematics education.

]]> Rethinking Mediation in Early Childhood Mathematics: A Conceptual Framework Informed by Inclusive Research Francesca Granone doi: 10.3390/proceedings2026141002 Proceedings 2026-06-29 Proceedings 2026-06-29 141 1 Proceeding Paper 2 10.3390/proceedings2026141002 https://www.mdpi.com/2504-3900/141/1/2 Proceedings, Vol. 146, Pages 122: Spatial Hotspots, Habitat Partitioning and Seasonal Dynamics of Sharks and Batoids in Lhaviyani Atoll, Central Maldives https://www.mdpi.com/2504-3900/146/1/122 Introduction: As apex and mesopredators, elasmobranchs help maintain marine ecosystem balance by shaping food-web structure and habitat connectivity, yet more than one-third of species are threatened with extinction. Identifying where and when they aggregate within atoll systems is therefore a prerequisite for spatially explicit conservation planning. Lhaviyani Atoll, in the central Maldives, lies within a recognised Indian Ocean elasmobranch hotspot and hosts two Important Shark and Ray Areas (ISRAs), yet fine-scale information on aggregation sites, habitat partitioning and seasonal use remains limited. Objective: To map persistent activity hotspots, characterise habitat partitioning between sharks and batoids, quantify seasonal and inter-annual dynamics, and provide an ecological basis for habitat-focused conservation in Lhaviyani Atoll. Methodology: Using a seven-year (2017–2024) opportunistic dive-log dataset of 12,732 SCUBA surveys and 142,994 elasmobranch records across 94 dive sites, spatial kernel-density estimation was applied separately to sharks and batoids to identify activity hotspots and visualise spatial overlap. Habitat associations were examined across substrate types and reef geomorphic zones. Seasonal and inter-annual dynamics in relative abundance and diversity (Shannon, Pielou’s evenness) were quantified across monsoon phases and the 2017–2024 period. Results: Twenty-eight species (14 sharks, 14 batoids) were recorded, including 23 listed as threatened on the IUCN Red List (4 Critically Endangered, 12 Endangered, 7 Vulnerable). Four persistent activity hotspots were identified along the northern atoll rim, two overlapping with the Fushifaru Kandu and Kuredhu–Huravalhi–Komandoo ISRAs. Sharks were concentrated along more complex exposed and semi-sheltered slopes and high-flow channels, with significantly higher occurrence on reef and sheltered reef slopes and lower occurrence on rubble and sand substrates; batoids were distributed broadly within lagoonal habitats with no strong substrate or geomorphic preferences. Relative abundance and diversity peaked during the late southwest monsoon (August–September) and declined into the northeast monsoon (December–March); after 2021, diversity and evenness increased while overall abundance declined. Conclusions: Persistent hotspots, contrasting habitat use by sharks and batoids, and consistent monsoonal seasonality support the ecological relevance of existing ISRAs in Lhaviyani Atoll, while providing finer-scale information on habitat partitioning and additional priority areas for threatened elasmobranchs, including four Critically Endangered species. 2026-06-25 Proceedings, Vol. 146, Pages 122: Spatial Hotspots, Habitat Partitioning and Seasonal Dynamics of Sharks and Batoids in Lhaviyani Atoll, Central Maldives

Proceedings doi: 10.3390/proceedings2026146122

Authors: Margarida Vizeu-Pinheiro Sebastião Farias Maria Lourie Saoirse Tak-Yung Macklin Paula Dominguez Rein-Loring Ray van Eeden Rui Rosa

Introduction: As apex and mesopredators, elasmobranchs help maintain marine ecosystem balance by shaping food-web structure and habitat connectivity, yet more than one-third of species are threatened with extinction. Identifying where and when they aggregate within atoll systems is therefore a prerequisite for spatially explicit conservation planning. Lhaviyani Atoll, in the central Maldives, lies within a recognised Indian Ocean elasmobranch hotspot and hosts two Important Shark and Ray Areas (ISRAs), yet fine-scale information on aggregation sites, habitat partitioning and seasonal use remains limited. Objective: To map persistent activity hotspots, characterise habitat partitioning between sharks and batoids, quantify seasonal and inter-annual dynamics, and provide an ecological basis for habitat-focused conservation in Lhaviyani Atoll. Methodology: Using a seven-year (2017–2024) opportunistic dive-log dataset of 12,732 SCUBA surveys and 142,994 elasmobranch records across 94 dive sites, spatial kernel-density estimation was applied separately to sharks and batoids to identify activity hotspots and visualise spatial overlap. Habitat associations were examined across substrate types and reef geomorphic zones. Seasonal and inter-annual dynamics in relative abundance and diversity (Shannon, Pielou’s evenness) were quantified across monsoon phases and the 2017–2024 period. Results: Twenty-eight species (14 sharks, 14 batoids) were recorded, including 23 listed as threatened on the IUCN Red List (4 Critically Endangered, 12 Endangered, 7 Vulnerable). Four persistent activity hotspots were identified along the northern atoll rim, two overlapping with the Fushifaru Kandu and Kuredhu–Huravalhi–Komandoo ISRAs. Sharks were concentrated along more complex exposed and semi-sheltered slopes and high-flow channels, with significantly higher occurrence on reef and sheltered reef slopes and lower occurrence on rubble and sand substrates; batoids were distributed broadly within lagoonal habitats with no strong substrate or geomorphic preferences. Relative abundance and diversity peaked during the late southwest monsoon (August–September) and declined into the northeast monsoon (December–March); after 2021, diversity and evenness increased while overall abundance declined. Conclusions: Persistent hotspots, contrasting habitat use by sharks and batoids, and consistent monsoonal seasonality support the ecological relevance of existing ISRAs in Lhaviyani Atoll, while providing finer-scale information on habitat partitioning and additional priority areas for threatened elasmobranchs, including four Critically Endangered species.

]]> Spatial Hotspots, Habitat Partitioning and Seasonal Dynamics of Sharks and Batoids in Lhaviyani Atoll, Central Maldives Margarida Vizeu-Pinheiro Sebastião Farias Maria Lourie Saoirse Tak-Yung Macklin Paula Dominguez Rein-Loring Ray van Eeden Rui Rosa doi: 10.3390/proceedings2026146122 Proceedings 2026-06-25 Proceedings 2026-06-25 146 1 Abstract 122 10.3390/proceedings2026146122 https://www.mdpi.com/2504-3900/146/1/122 Proceedings, Vol. 146, Pages 120: eDNA Metabarcoding and Traditional Surveys for Fish Monitoring in Coastal Wetlands https://www.mdpi.com/2504-3900/146/1/120 Introduction: Mediterranean coastal wetlands are highly dynamic ecosystems that support diverse fish communities and are often of high conservation value. The Ebro Delta is one of the most important coastal wetlands in the Western Mediterranean, and knowledge of fish assemblages is essential for its effective conservation and management. Environmental DNA (eDNA) metabarcoding provides a non-invasive approach that can potentially complement conventional surveys for fish biodiversity monitoring. Objective: This study aimed to evaluate the potential of eDNA metabarcoding as a complementary tool to conventional fyke net surveys for fish biodiversity monitoring in the Mediterranean coastal wetlands. Methodology: In 2022, fish assemblages were surveyed across 12 areas of the Ebro Delta using eDNA metabarcoding (12S MiFish) and conventional fyke net sampling. Results were compared with a 22-year historical dataset. Results: A total of 27 fish taxa were detected, 13 of which were exclusive to eDNA, 11 were shared between methods, and three were recorded only by fyke nets. The reliability of eDNA metabarcoding was supported by the detection of endangered species, such as Anguilla anguilla and Apricaphanius iberus; ubiquitous taxa, such as Atherina boyeri and Pomatoschistus microps; and invasive species, such as Gambusia holbrooki and Cyprinus carpio. Detection of invasive species was maximized using eDNA. While eDNA revealed higher species richness than fyke nets, community composition differed significantly between methods, reflecting distinct detection patterns. eDNA preferentially detected non-benthic species, whereas fyke nets were more robust for benthic taxa detection. eDNA recovered most historically recorded species but failed to detect some taxa, such as Misgurnus anguillicaudatus and Sardina pilchardus. Despite richness differences, the two methods provided complementary views of fish assemblages, highlighting method-specific detection limitations and opportunities. Conclusions: eDNA does not fully replace conventional surveys and their combined use improves the detection of threatened and invasive species, better supporting conservation and management. 2026-06-23 Proceedings, Vol. 146, Pages 120: eDNA Metabarcoding and Traditional Surveys for Fish Monitoring in Coastal Wetlands

Proceedings doi: 10.3390/proceedings2026146120

Authors: Nati Franch Marc Ventura Carles Alcaraz Víctor Osorio David Mateu Lluís Jornet Helena Fanlo Josep M. Queral Miguel Clavero Núria Cid

Introduction: Mediterranean coastal wetlands are highly dynamic ecosystems that support diverse fish communities and are often of high conservation value. The Ebro Delta is one of the most important coastal wetlands in the Western Mediterranean, and knowledge of fish assemblages is essential for its effective conservation and management. Environmental DNA (eDNA) metabarcoding provides a non-invasive approach that can potentially complement conventional surveys for fish biodiversity monitoring. Objective: This study aimed to evaluate the potential of eDNA metabarcoding as a complementary tool to conventional fyke net surveys for fish biodiversity monitoring in the Mediterranean coastal wetlands. Methodology: In 2022, fish assemblages were surveyed across 12 areas of the Ebro Delta using eDNA metabarcoding (12S MiFish) and conventional fyke net sampling. Results were compared with a 22-year historical dataset. Results: A total of 27 fish taxa were detected, 13 of which were exclusive to eDNA, 11 were shared between methods, and three were recorded only by fyke nets. The reliability of eDNA metabarcoding was supported by the detection of endangered species, such as Anguilla anguilla and Apricaphanius iberus; ubiquitous taxa, such as Atherina boyeri and Pomatoschistus microps; and invasive species, such as Gambusia holbrooki and Cyprinus carpio. Detection of invasive species was maximized using eDNA. While eDNA revealed higher species richness than fyke nets, community composition differed significantly between methods, reflecting distinct detection patterns. eDNA preferentially detected non-benthic species, whereas fyke nets were more robust for benthic taxa detection. eDNA recovered most historically recorded species but failed to detect some taxa, such as Misgurnus anguillicaudatus and Sardina pilchardus. Despite richness differences, the two methods provided complementary views of fish assemblages, highlighting method-specific detection limitations and opportunities. Conclusions: eDNA does not fully replace conventional surveys and their combined use improves the detection of threatened and invasive species, better supporting conservation and management.

]]> eDNA Metabarcoding and Traditional Surveys for Fish Monitoring in Coastal Wetlands Nati Franch Marc Ventura Carles Alcaraz Víctor Osorio David Mateu Lluís Jornet Helena Fanlo Josep M. Queral Miguel Clavero Núria Cid doi: 10.3390/proceedings2026146120 Proceedings 2026-06-23 Proceedings 2026-06-23 146 1 Abstract 120 10.3390/proceedings2026146120 https://www.mdpi.com/2504-3900/146/1/120 Proceedings, Vol. 146, Pages 121: Aglomerular Renal Function in Teleosts: A Comparative Molecular and Physiological Approach https://www.mdpi.com/2504-3900/146/1/121 Introduction: The aglomerular kidney, characterised by the absence of functional glomeruli and reliance on tubular secretion alone, has evolved independently across multiple teleost lineages occupying diverse environments, including notothenioids of the Southern Ocean, gadids of cold North Atlantic and Arctic waters, and syngnathids distributed across temperate and tropical seas. Despite sharing this independently derived renal architecture, these groups face markedly distinct osmotic, thermal and chemical challenges in their natural habitats. How aglomerular kidneys cope with environmental stressors, including fluctuations in temperature and salinity, and how they handle the excretion of xenobiotics and other exogenous compounds through exclusively tubular mechanisms, remains poorly understood. The physiological and molecular responses underlying renal function in these lineages have received comparatively little attention relative to their glomerular counterparts. Objective: This study investigates how aglomerular kidneys across phylogenetically distinct teleost lineages respond, at molecular and physiological levels, to contrasting environmental conditions, namely temperature and salinity challenges predicted to alter the functional demands on renal osmoregulation. Methodology: Adult and juvenile specimens from target lineages were subjected to controlled exposure experiments combining different temperature and salinity regimes. Blood and urine samples were collected to assess osmolality and ionic composition. Renal tissues were processed for gene expression analysis of key transport and structural genes, histology, immunohistochemistry and enzymatic activity assays, providing an integrated picture of renal function under each condition. Results: Molecular analyses are currently underway. Preliminary work has established protocols for RNA extraction and quantitative PCR across target species, enabling comparative gene expression profiling to proceed across the full dataset. Conclusions: A comparative physiological and molecular approach across aglomerular teleost lineages will clarify whether shared renal architecture translates into shared functional responses to environmental challenge and identify lineage-specific mechanisms of renal adaptation, with broader relevance for predicting how these fishes may respond to ongoing environmental change. 2026-06-23 Proceedings, Vol. 146, Pages 121: Aglomerular Renal Function in Teleosts: A Comparative Molecular and Physiological Approach

Proceedings doi: 10.3390/proceedings2026146121

Authors: José Teixeira Bernardo Pinto Jonathan M. Wilson Pedro M. Guerreiro Filipe Castro

Introduction: The aglomerular kidney, characterised by the absence of functional glomeruli and reliance on tubular secretion alone, has evolved independently across multiple teleost lineages occupying diverse environments, including notothenioids of the Southern Ocean, gadids of cold North Atlantic and Arctic waters, and syngnathids distributed across temperate and tropical seas. Despite sharing this independently derived renal architecture, these groups face markedly distinct osmotic, thermal and chemical challenges in their natural habitats. How aglomerular kidneys cope with environmental stressors, including fluctuations in temperature and salinity, and how they handle the excretion of xenobiotics and other exogenous compounds through exclusively tubular mechanisms, remains poorly understood. The physiological and molecular responses underlying renal function in these lineages have received comparatively little attention relative to their glomerular counterparts. Objective: This study investigates how aglomerular kidneys across phylogenetically distinct teleost lineages respond, at molecular and physiological levels, to contrasting environmental conditions, namely temperature and salinity challenges predicted to alter the functional demands on renal osmoregulation. Methodology: Adult and juvenile specimens from target lineages were subjected to controlled exposure experiments combining different temperature and salinity regimes. Blood and urine samples were collected to assess osmolality and ionic composition. Renal tissues were processed for gene expression analysis of key transport and structural genes, histology, immunohistochemistry and enzymatic activity assays, providing an integrated picture of renal function under each condition. Results: Molecular analyses are currently underway. Preliminary work has established protocols for RNA extraction and quantitative PCR across target species, enabling comparative gene expression profiling to proceed across the full dataset. Conclusions: A comparative physiological and molecular approach across aglomerular teleost lineages will clarify whether shared renal architecture translates into shared functional responses to environmental challenge and identify lineage-specific mechanisms of renal adaptation, with broader relevance for predicting how these fishes may respond to ongoing environmental change.

]]> Aglomerular Renal Function in Teleosts: A Comparative Molecular and Physiological Approach José Teixeira Bernardo Pinto Jonathan M. Wilson Pedro M. Guerreiro Filipe Castro doi: 10.3390/proceedings2026146121 Proceedings 2026-06-23 Proceedings 2026-06-23 146 1 Abstract 121 10.3390/proceedings2026146121 https://www.mdpi.com/2504-3900/146/1/121 Proceedings, Vol. 146, Pages 116: Life-History Parameters and Population Dynamics of Key Small Pelagic Fishes in São Tomé and Príncipe (Gulf of Guinea) https://www.mdpi.com/2504-3900/146/1/116 Small pelagic fishes are essential for artisanal fisheries and food security in São Tomé and Príncipe, yet biological information required for stock assessment remains scarce. This study examined the population dynamics and life-history traits of Caranx crysos, Euthynnus alletteratus, Hemiramphus balao, and Cheilopogon melanurus using 9052 specimens collected from artisanal landings between 2023 and 2025. C. melanurus (35.2%) and H. balao (34.0%) dominated the sampled catches, followed by C. crysos (18.1%) and E. alletteratus (12.7%). Standardized CPUE series modelled using GAMs revealed significant temporal and seasonal variation in relative abundance, with contrasting species-specific trends. Length–weight relationships revealed negative allometric growth in three of the four species examined (75%), with b values ranging from 2.50 to 3.19, while Fulton’s condition factor differed significantly among species (Kruskal–Wallis χ2 = 6702.7, p < 0.001). Sex-ratio analyses showed significant deviations from parity in C. crysos and C. melanurus, whereas E. alletteratus and H. balao remained balanced. Gonadosomatic index and maturity-stage distributions indicated year-round reproductive activity with distinct spawning peaks. Length at first maturity (L50) ranged from 30.2 cm to 38.8 cm among species. Growth parameters estimated from length-frequency data using the von Bertalanffy Growth Function fitted through ELEFAN_GA in TropFishR yielded L∞ values of 43.9–68.4 cm and K values of 0.065–0.336 yr⁻1. Growth performance index (φ′) ranged from 2.48 to 2.99, corresponding to theoretical longevities of 8.9–46.3 years. Length-based cohort analysis indicated biomass concentration in intermediate size classes and increasing fishing mortality towards larger individuals. Exploitation rates revealed contrasting fishing pressures among species, while sensitivity analyses identified growth and mortality parameters as the main sources of uncertainty. These findings provide the first integrated biological baseline for the assessment and management of small pelagic resources in São Tomé and Príncipe. 2026-06-23 Proceedings, Vol. 146, Pages 116: Life-History Parameters and Population Dynamics of Key Small Pelagic Fishes in São Tomé and Príncipe (Gulf of Guinea)

Proceedings doi: 10.3390/proceedings2026146116

Authors: Wilfred Boa Morte Zacarias Bupebe Júlio Sanca Mirian Gorett Gomes Cravid Bocar Sabaly Baldé

Small pelagic fishes are essential for artisanal fisheries and food security in São Tomé and Príncipe, yet biological information required for stock assessment remains scarce. This study examined the population dynamics and life-history traits of Caranx crysos, Euthynnus alletteratus, Hemiramphus balao, and Cheilopogon melanurus using 9052 specimens collected from artisanal landings between 2023 and 2025. C. melanurus (35.2%) and H. balao (34.0%) dominated the sampled catches, followed by C. crysos (18.1%) and E. alletteratus (12.7%). Standardized CPUE series modelled using GAMs revealed significant temporal and seasonal variation in relative abundance, with contrasting species-specific trends. Length–weight relationships revealed negative allometric growth in three of the four species examined (75%), with b values ranging from 2.50 to 3.19, while Fulton’s condition factor differed significantly among species (Kruskal–Wallis χ2 = 6702.7, p < 0.001). Sex-ratio analyses showed significant deviations from parity in C. crysos and C. melanurus, whereas E. alletteratus and H. balao remained balanced. Gonadosomatic index and maturity-stage distributions indicated year-round reproductive activity with distinct spawning peaks. Length at first maturity (L50) ranged from 30.2 cm to 38.8 cm among species. Growth parameters estimated from length-frequency data using the von Bertalanffy Growth Function fitted through ELEFAN_GA in TropFishR yielded L∞ values of 43.9–68.4 cm and K values of 0.065–0.336 yr⁻1. Growth performance index (φ′) ranged from 2.48 to 2.99, corresponding to theoretical longevities of 8.9–46.3 years. Length-based cohort analysis indicated biomass concentration in intermediate size classes and increasing fishing mortality towards larger individuals. Exploitation rates revealed contrasting fishing pressures among species, while sensitivity analyses identified growth and mortality parameters as the main sources of uncertainty. These findings provide the first integrated biological baseline for the assessment and management of small pelagic resources in São Tomé and Príncipe.

]]> Life-History Parameters and Population Dynamics of Key Small Pelagic Fishes in São Tomé and Príncipe (Gulf of Guinea) Wilfred Boa Morte Zacarias Bupebe Júlio Sanca Mirian Gorett Gomes Cravid Bocar Sabaly Baldé doi: 10.3390/proceedings2026146116 Proceedings 2026-06-23 Proceedings 2026-06-23 146 1 Abstract 116 10.3390/proceedings2026146116 https://www.mdpi.com/2504-3900/146/1/116 Proceedings, Vol. 146, Pages 119: LIFE RESQUE ALPYR: Ecological Restoration of High Mountain Lakes in the Pyrenees by Fish Removal https://www.mdpi.com/2504-3900/146/1/119 Currently, more than half of Pyrenean high mountain lakes are occupied by fish as a result of historical introductions that date back centuries and which have accelerated during the last 70 years. In the southern slope of these mountains, the main fish that have been introduced are Brown trout (Salmo trutta), Brook trout (Salvelinus fontinalis), Rainbow trout (Oncorhynchus mykiss), and European minnow (Phoxinus sp). The specific impacts of the introduction of fish include, among others, the transformation of the ecosystem structure and trophic relationships, and the reduction and extirpation of native species. The project LIFE RESQUE ALPYR (LIFE20 NAT/ES/000369), started in 2022 and ending in 2026, includes among its main objectives the restoration of ten high mountain lakes with fish (trout or minnow) and the recovery of native species of European interest by the eradication of introduced fish. We planned and executed continuous and sustained campaigns to achieve the complete removal of fish. From 2022, we began with operations in seven objective lakes by means of several capture techniques, mainly gill nets for trout and a combination of gill nets, fyke-nets, and electrofishing for minnows. In 2024 and 2025, in three other lakes, chemical treatments with rotenone were carried out to achieve rapid eradication of fish. Currently, we have already achieved the complete removal of fish in four lakes, either with sustained capture or chemical treatments. In the other three lakes, this objective is also expected to be achieved in 2026, and only few individuals persist in actuality. In the other two lakes, the European minnow has been removed, and trout are now the focus of a two-stage strategy. In the remaining lake, we have only achieved a reduction in the European minnow (>50% reduction), with trout still remaining. We present, in detail, the methodologies applied and the results obtained. 2026-06-23 Proceedings, Vol. 146, Pages 119: LIFE RESQUE ALPYR: Ecological Restoration of High Mountain Lakes in the Pyrenees by Fish Removal

Proceedings doi: 10.3390/proceedings2026146119

Authors: Quim Pou-Rovira Jordi Delgado Eloi Cruset Teresa Buchaca Víctor Osorio Danilo Buñay Nerina Gilbert Claudia Riera Barend Vandrooge Raimon Prats Pilar Fernández Joan O. Grimalt Rocco Tiberti Marc Ventura

Currently, more than half of Pyrenean high mountain lakes are occupied by fish as a result of historical introductions that date back centuries and which have accelerated during the last 70 years. In the southern slope of these mountains, the main fish that have been introduced are Brown trout (Salmo trutta), Brook trout (Salvelinus fontinalis), Rainbow trout (Oncorhynchus mykiss), and European minnow (Phoxinus sp). The specific impacts of the introduction of fish include, among others, the transformation of the ecosystem structure and trophic relationships, and the reduction and extirpation of native species. The project LIFE RESQUE ALPYR (LIFE20 NAT/ES/000369), started in 2022 and ending in 2026, includes among its main objectives the restoration of ten high mountain lakes with fish (trout or minnow) and the recovery of native species of European interest by the eradication of introduced fish. We planned and executed continuous and sustained campaigns to achieve the complete removal of fish. From 2022, we began with operations in seven objective lakes by means of several capture techniques, mainly gill nets for trout and a combination of gill nets, fyke-nets, and electrofishing for minnows. In 2024 and 2025, in three other lakes, chemical treatments with rotenone were carried out to achieve rapid eradication of fish. Currently, we have already achieved the complete removal of fish in four lakes, either with sustained capture or chemical treatments. In the other three lakes, this objective is also expected to be achieved in 2026, and only few individuals persist in actuality. In the other two lakes, the European minnow has been removed, and trout are now the focus of a two-stage strategy. In the remaining lake, we have only achieved a reduction in the European minnow (>50% reduction), with trout still remaining. We present, in detail, the methodologies applied and the results obtained.

]]> LIFE RESQUE ALPYR: Ecological Restoration of High Mountain Lakes in the Pyrenees by Fish Removal Quim Pou-Rovira Jordi Delgado Eloi Cruset Teresa Buchaca Víctor Osorio Danilo Buñay Nerina Gilbert Claudia Riera Barend Vandrooge Raimon Prats Pilar Fernández Joan O. Grimalt Rocco Tiberti Marc Ventura doi: 10.3390/proceedings2026146119 Proceedings 2026-06-23 Proceedings 2026-06-23 146 1 Abstract 119 10.3390/proceedings2026146119 https://www.mdpi.com/2504-3900/146/1/119 Proceedings, Vol. 146, Pages 114: Otolith Shape Variation in Bluefish (Pomatomus saltatrix) Along the Brazilian Southeast–South Coast Assessed Through Elliptical Fourier and Wavelet Transformed Descriptors https://www.mdpi.com/2504-3900/146/1/114 Introduction: Understanding population structure is essential for effective fishery management, and otolith shape analysis provides a robust framework for detecting spatial variation in marine fish populations. The bluefish (Pomatomus saltatrix), a commercially important and widely distributed species along the southeastern and southern Brazilian coast, may exhibit subtle population structuring that is still not fully resolved. Methodology: Otolith contour variation was analyzed using two complementary approaches: Elliptical Fourier Descriptors (EFD) and Wavelet Transformed Descriptors (WTD). A total of 75 individuals (25/site) were sampled from Rio de Janeiro, São Paulo, and Santa Catarina, with total lengths ranging from 33.2 to 45.5 cm. Multivariate analyses included Permutational Multivariate Analysis of Variance (PERMANOVA), pairwise Hotelling’s t-tests (HT2), Flexible Discriminant Analysis (FDA), and jackknife reclassification matrices (JKC). Results: For the EFD approach, PERMANOVA showed no significant differences among localities, while FDA revealed partial overlap among groups and a JKC overall reclassification accuracy of 46%. In contrast, the WTD approach detected significant spatial differences, with PERMANOVA indicating overall variation among localities and HT2 identifying significant differences between Santa Catarina and the other regions. FDA improved visual separation of Rio de Janeiro samples, although the JCK accuracy decreased to 35%. Conclusion: The combined results suggest the presence of weak to moderate spatial structuring in P. saltatrix along the studied coastline. However, inconsistencies among analytical approaches and relatively low reclassification success rate to the original site indicate that the observed differentiation is insufficient to conclusively define distinct population units, remaining compatible with either a single stock exhibiting spatial heterogeneity or weakly differentiated subpopulations. 2026-06-23 Proceedings, Vol. 146, Pages 114: Otolith Shape Variation in Bluefish (Pomatomus saltatrix) Along the Brazilian Southeast–South Coast Assessed Through Elliptical Fourier and Wavelet Transformed Descriptors

Proceedings doi: 10.3390/proceedings2026146114

Authors: Bianca Neves Felippe Alexandre Daros Rafael Schroeder Rafael Gaio Kulzer Marcus Rodrigues da Costa Rodolfo Miguel Silva Alberto Teodorico Correia

Introduction: Understanding population structure is essential for effective fishery management, and otolith shape analysis provides a robust framework for detecting spatial variation in marine fish populations. The bluefish (Pomatomus saltatrix), a commercially important and widely distributed species along the southeastern and southern Brazilian coast, may exhibit subtle population structuring that is still not fully resolved. Methodology: Otolith contour variation was analyzed using two complementary approaches: Elliptical Fourier Descriptors (EFD) and Wavelet Transformed Descriptors (WTD). A total of 75 individuals (25/site) were sampled from Rio de Janeiro, São Paulo, and Santa Catarina, with total lengths ranging from 33.2 to 45.5 cm. Multivariate analyses included Permutational Multivariate Analysis of Variance (PERMANOVA), pairwise Hotelling’s t-tests (HT2), Flexible Discriminant Analysis (FDA), and jackknife reclassification matrices (JKC). Results: For the EFD approach, PERMANOVA showed no significant differences among localities, while FDA revealed partial overlap among groups and a JKC overall reclassification accuracy of 46%. In contrast, the WTD approach detected significant spatial differences, with PERMANOVA indicating overall variation among localities and HT2 identifying significant differences between Santa Catarina and the other regions. FDA improved visual separation of Rio de Janeiro samples, although the JCK accuracy decreased to 35%. Conclusion: The combined results suggest the presence of weak to moderate spatial structuring in P. saltatrix along the studied coastline. However, inconsistencies among analytical approaches and relatively low reclassification success rate to the original site indicate that the observed differentiation is insufficient to conclusively define distinct population units, remaining compatible with either a single stock exhibiting spatial heterogeneity or weakly differentiated subpopulations.

]]> Otolith Shape Variation in Bluefish (Pomatomus saltatrix) Along the Brazilian Southeast–South Coast Assessed Through Elliptical Fourier and Wavelet Transformed Descriptors Bianca Neves Felippe Alexandre Daros Rafael Schroeder Rafael Gaio Kulzer Marcus Rodrigues da Costa Rodolfo Miguel Silva Alberto Teodorico Correia doi: 10.3390/proceedings2026146114 Proceedings 2026-06-23 Proceedings 2026-06-23 146 1 Abstract 114 10.3390/proceedings2026146114 https://www.mdpi.com/2504-3900/146/1/114 Proceedings, Vol. 146, Pages 113: Spermatozoa Morphology in Mediterranean Elasmobranchs https://www.mdpi.com/2504-3900/146/1/113 Introduction: Elasmobranchs play a crucial role in ecosystem regulation, but they are highly vulnerable to rapid environmental changes, particularly those driven by anthropogenic activities. Therefore, elasmobranchs are among the most threatened vertebrate groups worldwide, with overfishing and habitat degradation representing the primary threats to their survival. To address these challenges, in situ and ex situ conservation programs are complementary approaches. Objective: The implementation of assisted reproductive technologies, still poorly developed for elasmobranchs, represents a critical component of these ex situ strategies. Focused on that aspect, the main goal of this work was to get a better understanding of the sperm cells morphologies of different Mediterranean elasmobranch species. Results: The Elasmobranchii spermatozoa possesses a long and he-lical head, an elongated midpiece, and a flagellum supplemented with additional ultrastructural components to its axoneme. The comparative analysis of sperm head morphology revealed substantial interspecific variation among the studied elasmobranchs. Head length was relatively conserved, ranging from 48.5 to 62.0 μm, whereas helical parameters showed much greater variability. S. canicula and M. mobular exhibited the most compact head morphology, characterized by short helical wavelengths, low amplitudes, and the highest numbers of helices. In contrast, the batoids R. rhinobatos, R. radula, and R. clavata displayed broader, more widely spaced helices and fewer turns. Phylogenetic patterns were partially evident, as the closely related rajids shared very similar sperm morphology, while R. rhinobatos showed a comparable batoid morphotype. However, similarities between the distantly related M. mobular and S. canicula, and differences between the scyliorhinids S. canicula and G. melastomus, suggest that ecological and reproductive factors, in addition to phylogeny, have influenced the evolution of sperm head morphology in elasmobranchs. Conclusion: Elasmobranchii species possess big spermatozoa (compared to bony fishes) with an elongated helical head and tail similar to one currently existing (but later diverged) in birds, reptiles, and amphibians, which can be considered an evolutionary ancient. Sperm head morphology varies markedly among elasmobranchs, mainly regarding helical traits rather than head length. While phylogeny explains similarities among rajids, convergent patterns in distantly related species suggest that additional ecological and reproductive factors influence sperm evolution and structural design. 2026-06-23 Proceedings, Vol. 146, Pages 113: Spermatozoa Morphology in Mediterranean Elasmobranchs

Proceedings doi: 10.3390/proceedings2026146113

Authors: Yáiza F. Jorreto Victor Gallego Luz Pérez Thales S. França Juan F. Asturiano

Introduction: Elasmobranchs play a crucial role in ecosystem regulation, but they are highly vulnerable to rapid environmental changes, particularly those driven by anthropogenic activities. Therefore, elasmobranchs are among the most threatened vertebrate groups worldwide, with overfishing and habitat degradation representing the primary threats to their survival. To address these challenges, in situ and ex situ conservation programs are complementary approaches. Objective: The implementation of assisted reproductive technologies, still poorly developed for elasmobranchs, represents a critical component of these ex situ strategies. Focused on that aspect, the main goal of this work was to get a better understanding of the sperm cells morphologies of different Mediterranean elasmobranch species. Results: The Elasmobranchii spermatozoa possesses a long and he-lical head, an elongated midpiece, and a flagellum supplemented with additional ultrastructural components to its axoneme. The comparative analysis of sperm head morphology revealed substantial interspecific variation among the studied elasmobranchs. Head length was relatively conserved, ranging from 48.5 to 62.0 μm, whereas helical parameters showed much greater variability. S. canicula and M. mobular exhibited the most compact head morphology, characterized by short helical wavelengths, low amplitudes, and the highest numbers of helices. In contrast, the batoids R. rhinobatos, R. radula, and R. clavata displayed broader, more widely spaced helices and fewer turns. Phylogenetic patterns were partially evident, as the closely related rajids shared very similar sperm morphology, while R. rhinobatos showed a comparable batoid morphotype. However, similarities between the distantly related M. mobular and S. canicula, and differences between the scyliorhinids S. canicula and G. melastomus, suggest that ecological and reproductive factors, in addition to phylogeny, have influenced the evolution of sperm head morphology in elasmobranchs. Conclusion: Elasmobranchii species possess big spermatozoa (compared to bony fishes) with an elongated helical head and tail similar to one currently existing (but later diverged) in birds, reptiles, and amphibians, which can be considered an evolutionary ancient. Sperm head morphology varies markedly among elasmobranchs, mainly regarding helical traits rather than head length. While phylogeny explains similarities among rajids, convergent patterns in distantly related species suggest that additional ecological and reproductive factors influence sperm evolution and structural design.

]]> Spermatozoa Morphology in Mediterranean Elasmobranchs Yáiza F. Jorreto Victor Gallego Luz Pérez Thales S. França Juan F. Asturiano doi: 10.3390/proceedings2026146113 Proceedings 2026-06-23 Proceedings 2026-06-23 146 1 Abstract 113 10.3390/proceedings2026146113 https://www.mdpi.com/2504-3900/146/1/113 Proceedings, Vol. 146, Pages 117: Study of Exotic Ichthyofauna: The Particular Case of the Invasive Potential of Phoxinus phoxinus in Sousa River, North Portugal https://www.mdpi.com/2504-3900/146/1/117 Introduction: Biological invasions constitute one of the main threats to freshwater ecosystems, causing significant ecological changes through the introduction of exotic species that compete with or prey upon native species. In Portugal, the introduction and spread of exotic species in lotic and lentic ecosystems, such as pike (Esox lucius), European catfish (Silurus glanis), and largemouth bass (Micropterus salmoides), all top predators, may have a big impact on autochthonous species. In contrast, bleak (Alburnus alburnus), European perch (Perca fluviatilis), and common carp (Cyprinus carpio) compete aggressively for food resources. In the Sousa River basin, gudgeon (Gobio lozanoi) is considered an exotic species with potential ecological impact, with the minnow (Phoxinus phoxinus) stand having been recently identified in Portugal and, so far, recorded only in this river basin, and not yet being classified as an invasive species in Portugal. Public knowledge regarding invasive aquatic biodiversity remains a significant bottleneck for conservation. Because recreational angling is a prominent dispersal vector, initiatives that directly target this community are relevant. Objective: The aim is to carry out a bibliographic review on the exotic ichthyofauna species present in the Sousa River, with special focus on the invasion potential of the minnow (P. phoxinus). Methodology: The literature review was conducted based on the ScienceDirect, Springer Nature Link, and Fauna Norvegica databases, selecting publications between 2006 and 2025 concerning relevant studies on the potentially invasive characteristics of the minnow (P. phoxinus). The methodology is based on the analysis of studies regarding the impacts caused on riparian ecosystems. Results: The species P. phoxinus presents a generalist diet and high adaptive capacity, allowing it to colonise new habitats and compete aggressively with native species for trophic resources. Its presence is associated with negative impacts on brown trout populations (Salmo trutta), reducing growth and productivity, especially in mountain ecosystems. Increased species density also causes a significant decrease in benthic macroinvertebrate biodiversity. Studies conducted in the Douro basin indicate that the arrival of minnow in Portugal resulted from human action, probably associated with its use as live bait in recreational fishing. Conservation programmes use diverse tactics to bridge the awareness gap. Recent initiatives feature electrofishing demonstrations to visually differentiate species, theatrical performances, and even culinary show-cooking events using invasive predators like the European catfish to promote harvesting. Conclusions: The potential transition of P. phoxinus into an exotic and invasive species may be associated with the ecological pressure exerted on native communities, particularly through competition for trophic resources, highlighting the need to assess its dispersion in the Sousa basin and its impacts on fish fauna and benthic macroinvertebrates. It is important to do more sampling to understand its real distribution in the Sousa Basin. Additionally is important to explain to recreational anglers and the general population the impacts of fish transfer and the adverse effects of invasive species on freshwater Portuguese ecosystems. 2026-06-23 Proceedings, Vol. 146, Pages 117: Study of Exotic Ichthyofauna: The Particular Case of the Invasive Potential of Phoxinus phoxinus in Sousa River, North Portugal

Proceedings doi: 10.3390/proceedings2026146117

Authors: Hugo Lopes André Oliveira António Martinho João Soares Carrola

Introduction: Biological invasions constitute one of the main threats to freshwater ecosystems, causing significant ecological changes through the introduction of exotic species that compete with or prey upon native species. In Portugal, the introduction and spread of exotic species in lotic and lentic ecosystems, such as pike (Esox lucius), European catfish (Silurus glanis), and largemouth bass (Micropterus salmoides), all top predators, may have a big impact on autochthonous species. In contrast, bleak (Alburnus alburnus), European perch (Perca fluviatilis), and common carp (Cyprinus carpio) compete aggressively for food resources. In the Sousa River basin, gudgeon (Gobio lozanoi) is considered an exotic species with potential ecological impact, with the minnow (Phoxinus phoxinus) stand having been recently identified in Portugal and, so far, recorded only in this river basin, and not yet being classified as an invasive species in Portugal. Public knowledge regarding invasive aquatic biodiversity remains a significant bottleneck for conservation. Because recreational angling is a prominent dispersal vector, initiatives that directly target this community are relevant. Objective: The aim is to carry out a bibliographic review on the exotic ichthyofauna species present in the Sousa River, with special focus on the invasion potential of the minnow (P. phoxinus). Methodology: The literature review was conducted based on the ScienceDirect, Springer Nature Link, and Fauna Norvegica databases, selecting publications between 2006 and 2025 concerning relevant studies on the potentially invasive characteristics of the minnow (P. phoxinus). The methodology is based on the analysis of studies regarding the impacts caused on riparian ecosystems. Results: The species P. phoxinus presents a generalist diet and high adaptive capacity, allowing it to colonise new habitats and compete aggressively with native species for trophic resources. Its presence is associated with negative impacts on brown trout populations (Salmo trutta), reducing growth and productivity, especially in mountain ecosystems. Increased species density also causes a significant decrease in benthic macroinvertebrate biodiversity. Studies conducted in the Douro basin indicate that the arrival of minnow in Portugal resulted from human action, probably associated with its use as live bait in recreational fishing. Conservation programmes use diverse tactics to bridge the awareness gap. Recent initiatives feature electrofishing demonstrations to visually differentiate species, theatrical performances, and even culinary show-cooking events using invasive predators like the European catfish to promote harvesting. Conclusions: The potential transition of P. phoxinus into an exotic and invasive species may be associated with the ecological pressure exerted on native communities, particularly through competition for trophic resources, highlighting the need to assess its dispersion in the Sousa basin and its impacts on fish fauna and benthic macroinvertebrates. It is important to do more sampling to understand its real distribution in the Sousa Basin. Additionally is important to explain to recreational anglers and the general population the impacts of fish transfer and the adverse effects of invasive species on freshwater Portuguese ecosystems.

]]> Study of Exotic Ichthyofauna: The Particular Case of the Invasive Potential of Phoxinus phoxinus in Sousa River, North Portugal Hugo Lopes André Oliveira António Martinho João Soares Carrola doi: 10.3390/proceedings2026146117 Proceedings 2026-06-23 Proceedings 2026-06-23 146 1 Abstract 117 10.3390/proceedings2026146117 https://www.mdpi.com/2504-3900/146/1/117 Proceedings, Vol. 146, Pages 115: CMSY and Trend Analyses Reveal Depletion in Data-Poor Migratory Fish Stocks of Galicia (NW Spain) https://www.mdpi.com/2504-3900/146/1/115 Diadromous fishes support culturally important small-scale fisheries in Galicia (NW Spain), but their assessment is constrained by scarce biological and effort data. We evaluated nine data-poor fishery units using first-sale records for Alosa alosa, Anguilla anguilla, Petromyzon marinus and Platichthys flesus from six Galician ports (A Guarda, Arcade, Carril, Ferrol, Vigo and Rianxo) over the period 1997–2026. The workflow combined data cleaning and ecological classification, Mann–Kendall and Theil–Sen trend analyses, actual catch over maximum catch, and catch-only CMSY models with species-specific resilience priors. Results revealed widespread signals of depletion in both anadromous and catadromous groups. Six of the nine units were classified as collapsed or critical by CMSY, three as overexploited, and none as sustainable. The most severe cases were sea lamprey (P. marinus) in A Guarda and Carril (B/BMSY = 0.10 and 0.11, critical). A. alosa in A Guarda and European eel (A. anguilla) in Arcade, Carril and A Guarda (glass eel) were classified as collapsed (B/BMSY = 0.35–0.50). Trend analyses detected significant declines in lamprey at A Guarda and Carril and in adult eel at Ferrol, whereas P. flesus in Rianxo showed a significant positive slope but remained under alert status. Overall, three stocks showed increasing trends (although only one was statistically significant), while six showed declining trends (of which only three were significant). These findings showed that public landing time series can support precautionary assessment in data-limited migratory fisheries, and justify adaptive harvest restrictions, intensified monitoring, and routine screening to support recovery and management prioritization. 2026-06-23 Proceedings, Vol. 146, Pages 115: CMSY and Trend Analyses Reveal Depletion in Data-Poor Migratory Fish Stocks of Galicia (NW Spain)

Proceedings doi: 10.3390/proceedings2026146115

Authors: David J. Nachón Marta Cousido-Rocha Anxo Paz M. Grazia Pennino Santiago Cerviño Francisco Izquierdo Patricia Verísimo Pablo Caballero Estíbaliz Díaz

Diadromous fishes support culturally important small-scale fisheries in Galicia (NW Spain), but their assessment is constrained by scarce biological and effort data. We evaluated nine data-poor fishery units using first-sale records for Alosa alosa, Anguilla anguilla, Petromyzon marinus and Platichthys flesus from six Galician ports (A Guarda, Arcade, Carril, Ferrol, Vigo and Rianxo) over the period 1997–2026. The workflow combined data cleaning and ecological classification, Mann–Kendall and Theil–Sen trend analyses, actual catch over maximum catch, and catch-only CMSY models with species-specific resilience priors. Results revealed widespread signals of depletion in both anadromous and catadromous groups. Six of the nine units were classified as collapsed or critical by CMSY, three as overexploited, and none as sustainable. The most severe cases were sea lamprey (P. marinus) in A Guarda and Carril (B/BMSY = 0.10 and 0.11, critical). A. alosa in A Guarda and European eel (A. anguilla) in Arcade, Carril and A Guarda (glass eel) were classified as collapsed (B/BMSY = 0.35–0.50). Trend analyses detected significant declines in lamprey at A Guarda and Carril and in adult eel at Ferrol, whereas P. flesus in Rianxo showed a significant positive slope but remained under alert status. Overall, three stocks showed increasing trends (although only one was statistically significant), while six showed declining trends (of which only three were significant). These findings showed that public landing time series can support precautionary assessment in data-limited migratory fisheries, and justify adaptive harvest restrictions, intensified monitoring, and routine screening to support recovery and management prioritization.

]]> CMSY and Trend Analyses Reveal Depletion in Data-Poor Migratory Fish Stocks of Galicia (NW Spain) David J. Nachón Marta Cousido-Rocha Anxo Paz M. Grazia Pennino Santiago Cerviño Francisco Izquierdo Patricia Verísimo Pablo Caballero Estíbaliz Díaz doi: 10.3390/proceedings2026146115 Proceedings 2026-06-23 Proceedings 2026-06-23 146 1 Abstract 115 10.3390/proceedings2026146115 https://www.mdpi.com/2504-3900/146/1/115 Proceedings, Vol. 146, Pages 104: Monitoring the Use of Pelagic Fish Aggregation Devices by Largemouth Bass Using Tridimensional Fine-Scale Acoustic Positional Telemetry https://www.mdpi.com/2504-3900/146/1/104 Fish Aggregating Devices (FADs), traditionally used to attract and concentrate fish, can also serve as effective environmental enrichment tools in reservoirs, particularly in those with homogeneous characteristics and scarce refuge habitat, enhancing structural complexity and promoting recreational fishing opportunities. This study aimed to evaluate patterns in the use of prototype fish aggregation devices (FADs) in small size reservoirs. It was conducted at the Nascentes Reservoir (Crato), a small Mediterranean reservoir (ca. 10 ha) located in southern Portugal. These FADs were installed to enhance refuge habitat for fish species of interest to recreational fisheries, particularly largemouth bass (Micropterus salmoides Lacepède, 1802), thereby promoting the occurrence of trophy specimens. Two types of FADs were deployed and tested: (1) bank FADs (TREES), used in shallow waters near the margins; and (2) pelagic FADs (DAPs), suspended in the water column in deeper areas at the center of the reservoir. To monitor movement patterns and habitat use, an acoustic telemetry receiver array was deployed with a design to secure a three-dimensional fine-scale positioning with high accuracy. A total of 20 largemouth bass were tagged with acoustic transmitters equipped with pressure (i.e., depth) sensors. A before–after approach was used with 10 fish tracked before FAD deployment and 10 after. Results of fish behavior analysis provide strong evidence of fish using DAPs, but not TREES. In the presence of FADs, fish reduced their home ranges and movement amplitudes, becoming closely associated with these artificial habitats. Several environmental predictors explained fish behavior in the presence of artificial refuges, namely, diel period, moonlight intensity, and fish depth. The findings of this study are expected to contribute to the development of guidelines for refuge habitat enhancement in small- to medium-sized Mediterranean reservoirs, thereby increasing their recreational fishing attractiveness. 2026-06-23 Proceedings, Vol. 146, Pages 104: Monitoring the Use of Pelagic Fish Aggregation Devices by Largemouth Bass Using Tridimensional Fine-Scale Acoustic Positional Telemetry

Proceedings doi: 10.3390/proceedings2026146104

Authors: Miguel Encarnado Carlos M. Alexandre Bernardo Quintella Esmeralda Pereira Ana F. Belo Ana Filipa Silva João P. Marques António Faro Pedro R. Almeida

Fish Aggregating Devices (FADs), traditionally used to attract and concentrate fish, can also serve as effective environmental enrichment tools in reservoirs, particularly in those with homogeneous characteristics and scarce refuge habitat, enhancing structural complexity and promoting recreational fishing opportunities. This study aimed to evaluate patterns in the use of prototype fish aggregation devices (FADs) in small size reservoirs. It was conducted at the Nascentes Reservoir (Crato), a small Mediterranean reservoir (ca. 10 ha) located in southern Portugal. These FADs were installed to enhance refuge habitat for fish species of interest to recreational fisheries, particularly largemouth bass (Micropterus salmoides Lacepède, 1802), thereby promoting the occurrence of trophy specimens. Two types of FADs were deployed and tested: (1) bank FADs (TREES), used in shallow waters near the margins; and (2) pelagic FADs (DAPs), suspended in the water column in deeper areas at the center of the reservoir. To monitor movement patterns and habitat use, an acoustic telemetry receiver array was deployed with a design to secure a three-dimensional fine-scale positioning with high accuracy. A total of 20 largemouth bass were tagged with acoustic transmitters equipped with pressure (i.e., depth) sensors. A before–after approach was used with 10 fish tracked before FAD deployment and 10 after. Results of fish behavior analysis provide strong evidence of fish using DAPs, but not TREES. In the presence of FADs, fish reduced their home ranges and movement amplitudes, becoming closely associated with these artificial habitats. Several environmental predictors explained fish behavior in the presence of artificial refuges, namely, diel period, moonlight intensity, and fish depth. The findings of this study are expected to contribute to the development of guidelines for refuge habitat enhancement in small- to medium-sized Mediterranean reservoirs, thereby increasing their recreational fishing attractiveness.

]]> Monitoring the Use of Pelagic Fish Aggregation Devices by Largemouth Bass Using Tridimensional Fine-Scale Acoustic Positional Telemetry Miguel Encarnado Carlos M. Alexandre Bernardo Quintella Esmeralda Pereira Ana F. Belo Ana Filipa Silva João P. Marques António Faro Pedro R. Almeida doi: 10.3390/proceedings2026146104 Proceedings 2026-06-23 Proceedings 2026-06-23 146 1 Abstract 104 10.3390/proceedings2026146104 https://www.mdpi.com/2504-3900/146/1/104 Proceedings, Vol. 146, Pages 123: DiadSea Project: Transnational Cooperation to Improve the Management and Conservation of Diadromous Fish at Sea https://www.mdpi.com/2504-3900/146/1/123 Diadromous fish provide key ecological and socio-economic services in European Atlantic catchments, yet their marine phase remains poorly understood and weakly integrated into management. Involving nine partners from Portugal, Spain, France and Ireland, the DiadSea Interreg Atlantic Area initiative aims to fill these critical knowledge gaps on the marine and estuarine phases and to translate this information into coordinated conservation and fisheries management tools. To do so, the project combines historical and newly collected fishery-dependent and -independent data (landings, by-catch, cooperative surveys with commercial and recreational fishers) with advanced microchemical, genetic and environmental DNA (eDNA) analyses to characterize marine distributions, mixing areas and connectivity for shads, Atlantic salmon, sea trout, European eel, sea lamprey and other diadromous species. It also includes innovative case studies on lamprey tagging and intestinal metabarcoding, coastal habitat suitability mapping for shads using river plumes and environmental variables, and joint otolith microchemistry–genomics approaches to reassess European eel panmixia and maternal origin at the Atlantic scale. Species distribution models under present and future climate scenarios, specifically RCP4.5 and RCP8.5, are used to identify priority marine areas for conservation, zones of high temporal turnover and key interfaces ensuring longitudinal (river–sea) and latitudinal connectivity, which will feed into an updated interactive web Atlas of diadromous species. In parallel, DiadSea establishes a transnational observatory of stakeholders to harmonize legislation, co-develop adaptive fisheries management plans and produce climate-aware policy guidelines, while capacity-building actions include an origin-labeling scheme for sustainably harvested diadromous fish, educational games and a comic book to raise awareness among younger generations and the wider public. Together, these work packages will deliver the first integrated, marine-focused, evidence-based and decision-support framework for diadromous fishes in the North-Eastern Atlantic, strengthening conservation, sustainable fisheries and stakeholder engagement under ongoing climate change. 2026-06-22 Proceedings, Vol. 146, Pages 123: DiadSea Project: Transnational Cooperation to Improve the Management and Conservation of Diadromous Fish at Sea

Proceedings doi: 10.3390/proceedings2026146123

Authors: Rufino Vieira-Lanero Sandra Barca Fernando Cobo Catarina S. Mateus Pedro R. Almeida Joana Boavida-Portugal Carlos M. Alexandre Maria João Lança Helena Adão Bernardo Ruivo Quintella João Pereira Aurore Baisez Clarisse Boulenger Eric Feunteun Russell Poole Ciara O’Leary Anthony Brett

Diadromous fish provide key ecological and socio-economic services in European Atlantic catchments, yet their marine phase remains poorly understood and weakly integrated into management. Involving nine partners from Portugal, Spain, France and Ireland, the DiadSea Interreg Atlantic Area initiative aims to fill these critical knowledge gaps on the marine and estuarine phases and to translate this information into coordinated conservation and fisheries management tools. To do so, the project combines historical and newly collected fishery-dependent and -independent data (landings, by-catch, cooperative surveys with commercial and recreational fishers) with advanced microchemical, genetic and environmental DNA (eDNA) analyses to characterize marine distributions, mixing areas and connectivity for shads, Atlantic salmon, sea trout, European eel, sea lamprey and other diadromous species. It also includes innovative case studies on lamprey tagging and intestinal metabarcoding, coastal habitat suitability mapping for shads using river plumes and environmental variables, and joint otolith microchemistry–genomics approaches to reassess European eel panmixia and maternal origin at the Atlantic scale. Species distribution models under present and future climate scenarios, specifically RCP4.5 and RCP8.5, are used to identify priority marine areas for conservation, zones of high temporal turnover and key interfaces ensuring longitudinal (river–sea) and latitudinal connectivity, which will feed into an updated interactive web Atlas of diadromous species. In parallel, DiadSea establishes a transnational observatory of stakeholders to harmonize legislation, co-develop adaptive fisheries management plans and produce climate-aware policy guidelines, while capacity-building actions include an origin-labeling scheme for sustainably harvested diadromous fish, educational games and a comic book to raise awareness among younger generations and the wider public. Together, these work packages will deliver the first integrated, marine-focused, evidence-based and decision-support framework for diadromous fishes in the North-Eastern Atlantic, strengthening conservation, sustainable fisheries and stakeholder engagement under ongoing climate change.

]]> DiadSea Project: Transnational Cooperation to Improve the Management and Conservation of Diadromous Fish at Sea Rufino Vieira-Lanero Sandra Barca Fernando Cobo Catarina S. Mateus Pedro R. Almeida Joana Boavida-Portugal Carlos M. Alexandre Maria João Lança Helena Adão Bernardo Ruivo Quintella João Pereira Aurore Baisez Clarisse Boulenger Eric Feunteun Russell Poole Ciara O’Leary Anthony Brett doi: 10.3390/proceedings2026146123 Proceedings 2026-06-22 Proceedings 2026-06-22 146 1 Abstract 123 10.3390/proceedings2026146123 https://www.mdpi.com/2504-3900/146/1/123 Proceedings, Vol. 146, Pages 108: Transitional Waters: Critical Habitats for Coastal Fish Species and Fisheries † https://www.mdpi.com/2504-3900/146/1/108 Transitional waters—such as estuaries, lagoons, deltas, and coastal wetlands—are dynamic environments where freshwater and seawater interact, forming highly productive and biologically diverse ecosystems. Shaped by temperature and salinity gradients, tidal influence, sediment transport, and nutrient-rich conditions, these habitats support diverse ecological functions. Their structural complexity—including seagrass beds, salt marshes, mudflats, and mangroves—provides essential habitats for many fish species. These areas are crucial for fish life cycles, serving as nurseries, spawning grounds, feeding zones, and refuges from predators. Many commercially important species depend on them during early life stages before moving offshore, making them vital for both commercial and recreational fisheries. Beyond food provision, they deliver key ecosystem services, including water purification, coastal protection, and carbon storage. Research on the fish community of the Ria Formosa lagoon in Portugal since the 1980s highlights long-term changes in the fish community and the dominant role of habitat structure and temporal dynamics. Subtidal seagrass beds support higher fish abundance and diversity than unvegetated areas, acting as key nursery habitats and provide important fish provisioning services. Seasonal variation is also central, driven by recruitment pulses of marine migrants in late winter–spring. Recent pressures on this system have been driven by human activity and environmental change. Seagrass loss reduces nursery and feeding areas, while pollution degrades water quality. Overfishing (including illegal fishing), recreational activities, and aquaculture expansion add stress. Climate warming and invasive species such as Caulerpa prolifera, further disrupt ecosystem balance and threaten biodiversity. Sustainable management—such as habitat restoration, protected areas, and integrated policies—is essential to preserve the ecological and economic value of this unique lagoon. Ongoing research, monitoring, habitat restoration, and stakeholder engagement remain critical for ensuring resilience. 2026-06-22 Proceedings, Vol. 146, Pages 108: Transitional Waters: Critical Habitats for Coastal Fish Species and Fisheries †

Proceedings doi: 10.3390/proceedings2026146108

Authors: Karim Erzini

Transitional waters—such as estuaries, lagoons, deltas, and coastal wetlands—are dynamic environments where freshwater and seawater interact, forming highly productive and biologically diverse ecosystems. Shaped by temperature and salinity gradients, tidal influence, sediment transport, and nutrient-rich conditions, these habitats support diverse ecological functions. Their structural complexity—including seagrass beds, salt marshes, mudflats, and mangroves—provides essential habitats for many fish species. These areas are crucial for fish life cycles, serving as nurseries, spawning grounds, feeding zones, and refuges from predators. Many commercially important species depend on them during early life stages before moving offshore, making them vital for both commercial and recreational fisheries. Beyond food provision, they deliver key ecosystem services, including water purification, coastal protection, and carbon storage. Research on the fish community of the Ria Formosa lagoon in Portugal since the 1980s highlights long-term changes in the fish community and the dominant role of habitat structure and temporal dynamics. Subtidal seagrass beds support higher fish abundance and diversity than unvegetated areas, acting as key nursery habitats and provide important fish provisioning services. Seasonal variation is also central, driven by recruitment pulses of marine migrants in late winter–spring. Recent pressures on this system have been driven by human activity and environmental change. Seagrass loss reduces nursery and feeding areas, while pollution degrades water quality. Overfishing (including illegal fishing), recreational activities, and aquaculture expansion add stress. Climate warming and invasive species such as Caulerpa prolifera, further disrupt ecosystem balance and threaten biodiversity. Sustainable management—such as habitat restoration, protected areas, and integrated policies—is essential to preserve the ecological and economic value of this unique lagoon. Ongoing research, monitoring, habitat restoration, and stakeholder engagement remain critical for ensuring resilience.

]]> Transitional Waters: Critical Habitats for Coastal Fish Species and Fisheries † Karim Erzini doi: 10.3390/proceedings2026146108 Proceedings 2026-06-22 Proceedings 2026-06-22 146 1 Abstract 108 10.3390/proceedings2026146108 https://www.mdpi.com/2504-3900/146/1/108 Proceedings, Vol. 146, Pages 112: Preliminary Insights into the Daily and Seasonal Fishway Use by Luciobarbus bocagei Beyond the Spawning Migration Period https://www.mdpi.com/2504-3900/146/1/112 Introduction: River connectivity is a fundamental component of lotic ecosystems, frequently disrupted by anthropogenic barriers. Fishways are critical mitigation tools for restoring this connectivity; however, their temporal use by fish is often assumed to be restricted to specific seasonal windows (mainly spawning season). Understanding the fine-scale temporal dynamics of fishway use by native species such as the common barbel (Luciobarbus bocagei) is essential for effective, long-term river management and the conservation of Iberian freshwater biodiversity. Objective: This study aims to evaluate the daily and seasonal patterns of use of pool-type fishways by L. bocagei, in order to improve the determination of their temporal period of use and provide data-driven recommendations for their management. Methodology: The study was conducted in the middle reach of the Duero River (Burgos province). Over a multi-year period, more than 1000 individual barbels were captured, tagged with Passive Integrated Transponder (PIT) tags, and released. Movements were continuously monitored across two pool-type fishways. Telemetry detection data were analyzed to evaluate daily rhythms and seasonal use, as well as to estimate the start and end dates of the movement periods. Results: Our analysis revealed a highly marked daily pattern in fishway use, showing a clear increase during daylight hours, with more pronounced peaks at dawn and dusk. This daily activity dynamically adjusts throughout the year in response to the solar cycle. Furthermore, we successfully determined that the seasonal activity extends from March to November, a pattern that remained consistent throughout the different study years. Crucially, the telemetry data empirically demonstrated that L. bocagei utilizes fishways for a substantial portion of the year, indicating relevant non-reproductive movements. Conclusions: The extended temporal use of these fishways indicates their potential ecological functions throughout much of the year. This must be reflected in management and maintenance protocols that ensure sufficient flow conditions, cleaning, and active vigilance during most of the year. Additionally, this predictability reinforces the value of fishways as strategic biological monitoring stations for endemic populations. 2026-06-22 Proceedings, Vol. 146, Pages 112: Preliminary Insights into the Daily and Seasonal Fishway Use by Luciobarbus bocagei Beyond the Spawning Migration Period

Proceedings doi: 10.3390/proceedings2026146112

Authors: Francisco Javier Sanz-Ronda Juan Francisco Fuentes-Pérez Ana García-Vega Marina Martínez-Miguel Juan de María-Arnaiz Francisco Javier Bravo-Córdoba

Introduction: River connectivity is a fundamental component of lotic ecosystems, frequently disrupted by anthropogenic barriers. Fishways are critical mitigation tools for restoring this connectivity; however, their temporal use by fish is often assumed to be restricted to specific seasonal windows (mainly spawning season). Understanding the fine-scale temporal dynamics of fishway use by native species such as the common barbel (Luciobarbus bocagei) is essential for effective, long-term river management and the conservation of Iberian freshwater biodiversity. Objective: This study aims to evaluate the daily and seasonal patterns of use of pool-type fishways by L. bocagei, in order to improve the determination of their temporal period of use and provide data-driven recommendations for their management. Methodology: The study was conducted in the middle reach of the Duero River (Burgos province). Over a multi-year period, more than 1000 individual barbels were captured, tagged with Passive Integrated Transponder (PIT) tags, and released. Movements were continuously monitored across two pool-type fishways. Telemetry detection data were analyzed to evaluate daily rhythms and seasonal use, as well as to estimate the start and end dates of the movement periods. Results: Our analysis revealed a highly marked daily pattern in fishway use, showing a clear increase during daylight hours, with more pronounced peaks at dawn and dusk. This daily activity dynamically adjusts throughout the year in response to the solar cycle. Furthermore, we successfully determined that the seasonal activity extends from March to November, a pattern that remained consistent throughout the different study years. Crucially, the telemetry data empirically demonstrated that L. bocagei utilizes fishways for a substantial portion of the year, indicating relevant non-reproductive movements. Conclusions: The extended temporal use of these fishways indicates their potential ecological functions throughout much of the year. This must be reflected in management and maintenance protocols that ensure sufficient flow conditions, cleaning, and active vigilance during most of the year. Additionally, this predictability reinforces the value of fishways as strategic biological monitoring stations for endemic populations.

]]> Preliminary Insights into the Daily and Seasonal Fishway Use by Luciobarbus bocagei Beyond the Spawning Migration Period Francisco Javier Sanz-Ronda Juan Francisco Fuentes-Pérez Ana García-Vega Marina Martínez-Miguel Juan de María-Arnaiz Francisco Javier Bravo-Córdoba doi: 10.3390/proceedings2026146112 Proceedings 2026-06-22 Proceedings 2026-06-22 146 1 Abstract 112 10.3390/proceedings2026146112 https://www.mdpi.com/2504-3900/146/1/112 Proceedings, Vol. 146, Pages 97: Size-Based Indicators Reveal a Long-Term Decreasing Trend in an Estuarine Fish Assemblage and the Cumulative Impacts of Warming https://www.mdpi.com/2504-3900/146/1/97 Introduction: Long-term ecological changes in estuarine communities are primarily driven by anthropogenic and environmental pressures. While abundance-based indicators are commonly used to assess these shifts, they often mask underlying ecological aspects related to age and/or size dynamics that may not necessarily be reflected in the abundance-based approach. Objective: This work tested a size-based indicator approach to examine the long-term changes in the size structure of the Mondego estuarine fish community (Portugal), using a 22-year dataset (2003 to 2025). Methodology: To capture the whole size structure, eight size-based indicators were applied, including mean length (MeanL), length at the 10th percentile (L10), median length (MedianL), length at the 90th percentile (L90), mean length of the 90th percentile (Lmax), size spectrum, the Large Fish index, and the Shannon index of length classes, at community and species levels and subsequently considered these in relation with with local and large-scale environmental factors. Results: Linear models identified a sharp, consistent decline in the overall size of the community, significantly correlated with the North Atlantic Oscillation index (NAO) and increasing estuarine water temperatures. A dynamic factor analysis (DFA) further identified one common trend across species for all indicators, corroborating the decrease in the overall size of the community while also acknowledging contrasting responses from different species, suggesting a heterogenous response across the fish community. Conclusions: These results highlight the importance of size-based indicators when assessing long-term ecological changes in marine ecosystems, allowing us to better understand how size structures shift, their relationship with a changing environment, and the long-term ecological outcomes in terms of community stability, resilience, recruitment, and ecosystem functioning. 2026-06-22 Proceedings, Vol. 146, Pages 97: Size-Based Indicators Reveal a Long-Term Decreasing Trend in an Estuarine Fish Assemblage and the Cumulative Impacts of Warming

Proceedings doi: 10.3390/proceedings2026146097

Authors: Alexandre Carreira Sara Lourenço Manuel J. Rodrigues Filipe Costa Ana Lígia Primo Milene Guerreiro Miguel A. Pardal Szymon Smoliński Filipe Martinho

Introduction: Long-term ecological changes in estuarine communities are primarily driven by anthropogenic and environmental pressures. While abundance-based indicators are commonly used to assess these shifts, they often mask underlying ecological aspects related to age and/or size dynamics that may not necessarily be reflected in the abundance-based approach. Objective: This work tested a size-based indicator approach to examine the long-term changes in the size structure of the Mondego estuarine fish community (Portugal), using a 22-year dataset (2003 to 2025). Methodology: To capture the whole size structure, eight size-based indicators were applied, including mean length (MeanL), length at the 10th percentile (L10), median length (MedianL), length at the 90th percentile (L90), mean length of the 90th percentile (Lmax), size spectrum, the Large Fish index, and the Shannon index of length classes, at community and species levels and subsequently considered these in relation with with local and large-scale environmental factors. Results: Linear models identified a sharp, consistent decline in the overall size of the community, significantly correlated with the North Atlantic Oscillation index (NAO) and increasing estuarine water temperatures. A dynamic factor analysis (DFA) further identified one common trend across species for all indicators, corroborating the decrease in the overall size of the community while also acknowledging contrasting responses from different species, suggesting a heterogenous response across the fish community. Conclusions: These results highlight the importance of size-based indicators when assessing long-term ecological changes in marine ecosystems, allowing us to better understand how size structures shift, their relationship with a changing environment, and the long-term ecological outcomes in terms of community stability, resilience, recruitment, and ecosystem functioning.

]]> Size-Based Indicators Reveal a Long-Term Decreasing Trend in an Estuarine Fish Assemblage and the Cumulative Impacts of Warming Alexandre Carreira Sara Lourenço Manuel J. Rodrigues Filipe Costa Ana Lígia Primo Milene Guerreiro Miguel A. Pardal Szymon Smoliński Filipe Martinho doi: 10.3390/proceedings2026146097 Proceedings 2026-06-22 Proceedings 2026-06-22 146 1 Abstract 97 10.3390/proceedings2026146097 https://www.mdpi.com/2504-3900/146/1/97 Proceedings, Vol. 146, Pages 111: Longlines for Sampling, Reduction and Eradication of Large Alien Invasive Predatory Species: The Case of European Catfish https://www.mdpi.com/2504-3900/146/1/111 Introduction: Longlines are traditional fishing gear and are widely used in marine systems for both quantitative and qualitative sampling of large predators like tuna, swordfish, sailfish, marlin or sharks. This highly selective method has been applied for the sampling of European catfish (Silurus glanis), the largest freshwater fish in Europe. This apex predator is a highly valuable fish species in its native localities of Central and Eastern Europe (commercial fishing, anglers’ trophy fishing, biomanipulation purposes) but a dangerous alien invasive species in South and Western Europe. Objective: The efficiency and selectivity of longlines for European catfish sampling were compared with more traditional fishing methods like gillnets and electrofishing. Methodology: European catfish were sampled in native areas (Římov, Žlutice, Vrchlice, Hubenov, Lipno, Želivka reservoirs; Czech Republic) and areas of invasion (Lakes Campagna, Avigliana Grande, Maggiore; Italy; Belver, Meimoa, Cedillo reservoirs; Portugal; Iznajar, Mequinenza reservoirs; Spain) following the protocols of best catfish (Silurus glanis) capture methodologies in small and large lakes and reservoirs established for the current needs of the European Commission within the LIFE PREDATOR project. Results: The longline efficiency expressed as the proportion of individual hooks catching catfish (live baits exposed overnight) was from low to medium (5–25%; Czech reservoirs, Italian lakes, Meimoa reservoir) to very high (up to over 50%; Portuguese and Spanish reservoirs). In many water bodies, specifically in areas of invasion, catfish represented 100% of the longline catch (Campagna, Maggiore, Cedillo, Iznajar, Mequinenza). In the rest of the water bodies, the by-catch ranged from 2 to 20% with the highest values occurring in its native localities (due to the presence of large individuals of Northern pike, Esox lucius, another predatory fish in these systems). In areas of invasion, the inconsiderable by-catch was composed of other non-native predatory fish species like pikeperch (Sander lucioperca), European perch (Perca fluviatilis) or channel catfish (Ictalurus punctatus). Both the efficiency and selectivity of longlines were higher compared to gillnets and electrofishing, and longlines also caught larger catfish individuals. Conclusions: Longlines represent an efficient and extremely selective method for European catfish sampling, reduction and even eradication, especially in areas of invasion. Their usage, however, requires specific equipment, skills and also baits. 2026-06-22 Proceedings, Vol. 146, Pages 111: Longlines for Sampling, Reduction and Eradication of Large Alien Invasive Predatory Species: The Case of European Catfish

Proceedings doi: 10.3390/proceedings2026146111

Authors: Martin Čech Lukáš Vejřík Luboš Kočvara Vladislav Draštík Zuzana Sajdlová Diogo Dias Rui Rivaes Diogo Ribeiro Beatriz Castro Filipe Ribeiro Carlos Fernández-Delgado Agustín P. Monteoliva Pietro Volta

Introduction: Longlines are traditional fishing gear and are widely used in marine systems for both quantitative and qualitative sampling of large predators like tuna, swordfish, sailfish, marlin or sharks. This highly selective method has been applied for the sampling of European catfish (Silurus glanis), the largest freshwater fish in Europe. This apex predator is a highly valuable fish species in its native localities of Central and Eastern Europe (commercial fishing, anglers’ trophy fishing, biomanipulation purposes) but a dangerous alien invasive species in South and Western Europe. Objective: The efficiency and selectivity of longlines for European catfish sampling were compared with more traditional fishing methods like gillnets and electrofishing. Methodology: European catfish were sampled in native areas (Římov, Žlutice, Vrchlice, Hubenov, Lipno, Želivka reservoirs; Czech Republic) and areas of invasion (Lakes Campagna, Avigliana Grande, Maggiore; Italy; Belver, Meimoa, Cedillo reservoirs; Portugal; Iznajar, Mequinenza reservoirs; Spain) following the protocols of best catfish (Silurus glanis) capture methodologies in small and large lakes and reservoirs established for the current needs of the European Commission within the LIFE PREDATOR project. Results: The longline efficiency expressed as the proportion of individual hooks catching catfish (live baits exposed overnight) was from low to medium (5–25%; Czech reservoirs, Italian lakes, Meimoa reservoir) to very high (up to over 50%; Portuguese and Spanish reservoirs). In many water bodies, specifically in areas of invasion, catfish represented 100% of the longline catch (Campagna, Maggiore, Cedillo, Iznajar, Mequinenza). In the rest of the water bodies, the by-catch ranged from 2 to 20% with the highest values occurring in its native localities (due to the presence of large individuals of Northern pike, Esox lucius, another predatory fish in these systems). In areas of invasion, the inconsiderable by-catch was composed of other non-native predatory fish species like pikeperch (Sander lucioperca), European perch (Perca fluviatilis) or channel catfish (Ictalurus punctatus). Both the efficiency and selectivity of longlines were higher compared to gillnets and electrofishing, and longlines also caught larger catfish individuals. Conclusions: Longlines represent an efficient and extremely selective method for European catfish sampling, reduction and even eradication, especially in areas of invasion. Their usage, however, requires specific equipment, skills and also baits.

]]> Longlines for Sampling, Reduction and Eradication of Large Alien Invasive Predatory Species: The Case of European Catfish Martin Čech Lukáš Vejřík Luboš Kočvara Vladislav Draštík Zuzana Sajdlová Diogo Dias Rui Rivaes Diogo Ribeiro Beatriz Castro Filipe Ribeiro Carlos Fernández-Delgado Agustín P. Monteoliva Pietro Volta doi: 10.3390/proceedings2026146111 Proceedings 2026-06-22 Proceedings 2026-06-22 146 1 Abstract 111 10.3390/proceedings2026146111 https://www.mdpi.com/2504-3900/146/1/111 Proceedings, Vol. 146, Pages 96: Non-Native Inland Fish Across the Circum-Mediterranean Region: A Comprehensive Inventory https://www.mdpi.com/2504-3900/146/1/96 Introduction: The circum-Mediterranean region is a global biodiversity hotspot, hosting a highly distinctive freshwater fauna with a high degree of endemism and conservation concern. However, these ecosystems are increasingly threatened by biological invasions, particularly by non-native fish species, which represent a major driver of biodiversity loss. Objective: This study aims to compile a comprehensive and updated inventory of non-native inland fish species across the circum-Mediterranean region and to identify the main taxonomic, biogeographical, and socio-environmental drivers shaping their distribution. Methodology: We conducted an extensive review of the scientific literature, online databases (including EASIN, GISD, and CABI), and technical reports to compile records of non-native fish species across inland and transitional waters of Mediterranean-climate basins. Analyses focused on species composition, taxonomic representativeness, introduction pathways, native regions, and the relationship between species richness and selected environmental and socio-economic variables. Results: A total of 151 non-native fish species were recorded across the study area. Italy, Spain, Bosnia and Herzegovina, France, and Croatia exhibited the highest numbers of established species. Taxonomic representation was uneven, with Salmoniformes and Esociformes overrepresented among established non-native species, while Siluriformes and Characiformes were underrepresented. Most introductions originated from Europe, Asia, and North America, primarily through intentional releases and escape events. Non-native species richness was positively correlated with gross domestic product, precipitation, and the number of dams, highlighting the role of economic development and habitat modification in facilitating invasions. Conclusions: Biological invasions by non-native fishes are widespread across the Mediterranean basin and are strongly driven by human activities and environmental conditions. The high invasion levels observed in this biodiversity hotspot pose a significant threat to endemic freshwater faunas. These findings underscore the need for coordinated transnational management strategies, stricter regulation of introduction pathways, and prioritization of high-risk species to mitigate further impacts. 2026-06-22 Proceedings, Vol. 146, Pages 96: Non-Native Inland Fish Across the Circum-Mediterranean Region: A Comprehensive Inventory

Proceedings doi: 10.3390/proceedings2026146096

Authors: Carlos Cano-Barbacil Emili García-Berthou Filipe Ribeiro Marko Ćaleta Jesús Pedreño Francisco José Oliva-Paterna

Introduction: The circum-Mediterranean region is a global biodiversity hotspot, hosting a highly distinctive freshwater fauna with a high degree of endemism and conservation concern. However, these ecosystems are increasingly threatened by biological invasions, particularly by non-native fish species, which represent a major driver of biodiversity loss. Objective: This study aims to compile a comprehensive and updated inventory of non-native inland fish species across the circum-Mediterranean region and to identify the main taxonomic, biogeographical, and socio-environmental drivers shaping their distribution. Methodology: We conducted an extensive review of the scientific literature, online databases (including EASIN, GISD, and CABI), and technical reports to compile records of non-native fish species across inland and transitional waters of Mediterranean-climate basins. Analyses focused on species composition, taxonomic representativeness, introduction pathways, native regions, and the relationship between species richness and selected environmental and socio-economic variables. Results: A total of 151 non-native fish species were recorded across the study area. Italy, Spain, Bosnia and Herzegovina, France, and Croatia exhibited the highest numbers of established species. Taxonomic representation was uneven, with Salmoniformes and Esociformes overrepresented among established non-native species, while Siluriformes and Characiformes were underrepresented. Most introductions originated from Europe, Asia, and North America, primarily through intentional releases and escape events. Non-native species richness was positively correlated with gross domestic product, precipitation, and the number of dams, highlighting the role of economic development and habitat modification in facilitating invasions. Conclusions: Biological invasions by non-native fishes are widespread across the Mediterranean basin and are strongly driven by human activities and environmental conditions. The high invasion levels observed in this biodiversity hotspot pose a significant threat to endemic freshwater faunas. These findings underscore the need for coordinated transnational management strategies, stricter regulation of introduction pathways, and prioritization of high-risk species to mitigate further impacts.

]]> Non-Native Inland Fish Across the Circum-Mediterranean Region: A Comprehensive Inventory Carlos Cano-Barbacil Emili García-Berthou Filipe Ribeiro Marko Ćaleta Jesús Pedreño Francisco José Oliva-Paterna doi: 10.3390/proceedings2026146096 Proceedings 2026-06-22 Proceedings 2026-06-22 146 1 Abstract 96 10.3390/proceedings2026146096 https://www.mdpi.com/2504-3900/146/1/96 Proceedings, Vol. 146, Pages 110: Marine Recreational Fishing in Portugal: Social and Biological Perspectives for Improved Management https://www.mdpi.com/2504-3900/146/1/110 Introduction: Marine recreational fishing (MRF) is a socially, economically, and ecologically important activity. However, the absence of systematic monitoring and research in Portugal has limited a comprehensive understanding of the sector and its broader implications. Addressing these gaps is essential to support evidence-based and adaptive recreational fisheries management. Objective: This work aimed to improve understanding of marine recreational fishing in Portugal by integrating social and biological perspectives across national, regional, and local scales. Methodology: A multidisciplinary approach was applied. At the national scale, online questionnaire data were combined with licencing information and regional effort estimates to quantify fishing effort, catches, release rates, catch composition, and direct trip expenditures. The same survey framework was also used to identify recreational fisher profiles based on motivations, behaviours, fishing practices, and attitudes toward management. At the local scale, shore angling surveys conducted before the implementation of the Natural Marine Park of the Algarve Reef were used to estimate fishing activity and establish a baseline for future monitoring. From a biological perspective, field-based catch-and-release experiments were conducted to assess post-release mortality and physiological disturbance, using indicators such as mortality, condition, capture and handling variables, and blood biomarkers of stress and recovery. Results: The national estimates documented substantial fishing effort, catches, and direct expenditure, confirming the relevance of MRF in Portugal at both ecological and socio-economic levels. Portuguese recreational fishers were also shown to be heterogeneous in their motivations, behaviours, and practices, reinforcing the need for management approaches that reflect this diversity. At the local scale, shore angling estimates from the Natural Marine Park of the Algarve Reef provided an important pre-protection baseline for the now established Marine Protected Area, enabling future comparisons and long-term assessment of management effects. Biological experiments showed that recreational fishing can generate measurable physiological disturbance in fish, with post-release condition influenced by capture and handling factors. Among the variables examined, hook location emerged as particularly important, with deep-hooked fish showing greater impairment and higher mortality risk. Conclusions: Together, these findings show that improving recreational fisheries management in Portugal requires integrating social patterns, fishery magnitude, and biological responses to capture and release. Long-term monitoring programmes, combined with management approaches adapted to different fisher profiles and fishing contexts, are essential to support more effective and sustainable recreational fisheries governance. 2026-06-22 Proceedings, Vol. 146, Pages 110: Marine Recreational Fishing in Portugal: Social and Biological Perspectives for Improved Management

Proceedings doi: 10.3390/proceedings2026146110

Authors: Samira Nuñez-Velazquez Pedro M. Guerreiro Gonçalo Jacinto Pedro Veiga Mafalda Rangel

Introduction: Marine recreational fishing (MRF) is a socially, economically, and ecologically important activity. However, the absence of systematic monitoring and research in Portugal has limited a comprehensive understanding of the sector and its broader implications. Addressing these gaps is essential to support evidence-based and adaptive recreational fisheries management. Objective: This work aimed to improve understanding of marine recreational fishing in Portugal by integrating social and biological perspectives across national, regional, and local scales. Methodology: A multidisciplinary approach was applied. At the national scale, online questionnaire data were combined with licencing information and regional effort estimates to quantify fishing effort, catches, release rates, catch composition, and direct trip expenditures. The same survey framework was also used to identify recreational fisher profiles based on motivations, behaviours, fishing practices, and attitudes toward management. At the local scale, shore angling surveys conducted before the implementation of the Natural Marine Park of the Algarve Reef were used to estimate fishing activity and establish a baseline for future monitoring. From a biological perspective, field-based catch-and-release experiments were conducted to assess post-release mortality and physiological disturbance, using indicators such as mortality, condition, capture and handling variables, and blood biomarkers of stress and recovery. Results: The national estimates documented substantial fishing effort, catches, and direct expenditure, confirming the relevance of MRF in Portugal at both ecological and socio-economic levels. Portuguese recreational fishers were also shown to be heterogeneous in their motivations, behaviours, and practices, reinforcing the need for management approaches that reflect this diversity. At the local scale, shore angling estimates from the Natural Marine Park of the Algarve Reef provided an important pre-protection baseline for the now established Marine Protected Area, enabling future comparisons and long-term assessment of management effects. Biological experiments showed that recreational fishing can generate measurable physiological disturbance in fish, with post-release condition influenced by capture and handling factors. Among the variables examined, hook location emerged as particularly important, with deep-hooked fish showing greater impairment and higher mortality risk. Conclusions: Together, these findings show that improving recreational fisheries management in Portugal requires integrating social patterns, fishery magnitude, and biological responses to capture and release. Long-term monitoring programmes, combined with management approaches adapted to different fisher profiles and fishing contexts, are essential to support more effective and sustainable recreational fisheries governance.

]]> Marine Recreational Fishing in Portugal: Social and Biological Perspectives for Improved Management Samira Nuñez-Velazquez Pedro M. Guerreiro Gonçalo Jacinto Pedro Veiga Mafalda Rangel doi: 10.3390/proceedings2026146110 Proceedings 2026-06-22 Proceedings 2026-06-22 146 1 Abstract 110 10.3390/proceedings2026146110 https://www.mdpi.com/2504-3900/146/1/110 Proceedings, Vol. 146, Pages 109: Diagnosis of the Present Situation of the Spanish Toothcarp (Apricaphanius iberus) in Empordà Wetlands https://www.mdpi.com/2504-3900/146/1/109 Currently, in Catalonia, Spanish toothcarp (Apricaphanius iberus) has a discontinuous distribution, with its main populations located in the Ebro Delta and the Empordà wetlands. Along the Catalan coast, there are some small, isolated populations, most of which have been recently introduced. In the Empordà area, this species still maintains three isolated populations within two natural parks, where it currently occupies the northernmost site of its global distribution. Between 2017 and 2025, several fish sampling campaigns were carried out in the Empordà wetlands (Northeast Catalonia), gradually covering the entire area of potential distribution of this species in the area. In total, 228 points have been sampled, at least in one occasion, in all types of water bodies (rivers, canals, lagoons, marshes). In 106 points, more than two sampling events have been performed. Furthermore, in 2025, a specific diagnosis was carried out to better understand the current situation of the species in these protected spaces after a prolonged and severe drought. Sampling was carried out everywhere with fykenets, adding the use of nets and electrofishing in some stations located in large bodies of water. The known historical evolution of Spanish toothcarp distribution in the Empordà has been presented since the first data was made available at the beginning of the 1980s of the 20th century until now. Today, this fish only occupies approximately 10% of its potential distribution in the area. Over the last five decades, its distribution has expanded and contracted several times, but in 2025, it was once again in the worst possible situation of the analysis period. Therefore, the implementation of protected areas, or the execution of several large ecological restoration projects, have only succeeded in maintaining existing populations, but not in the expansion of its potential distribution to unoccupied areas. The main factors that explain this general situation are mainly (1) the proliferation of exotic species, (2) the loss of ecological status, and (3) hydrological alterations and increasingly severe droughts. Thus, the recovery and long-term conservation of Spanish toothcarp in the Empordà area inevitably requires the implementation of more extensive and decisive measures to reverse the incidence of these factors. 2026-06-22 Proceedings, Vol. 146, Pages 109: Diagnosis of the Present Situation of the Spanish Toothcarp (Apricaphanius iberus) in Empordà Wetlands

Proceedings doi: 10.3390/proceedings2026146109

Authors: Quim Pou-Rovira Neus Mairal Elena Farré Bernat Dalmau Llorenç Ferrer Maria Spotti Eloi Cruset

Currently, in Catalonia, Spanish toothcarp (Apricaphanius iberus) has a discontinuous distribution, with its main populations located in the Ebro Delta and the Empordà wetlands. Along the Catalan coast, there are some small, isolated populations, most of which have been recently introduced. In the Empordà area, this species still maintains three isolated populations within two natural parks, where it currently occupies the northernmost site of its global distribution. Between 2017 and 2025, several fish sampling campaigns were carried out in the Empordà wetlands (Northeast Catalonia), gradually covering the entire area of potential distribution of this species in the area. In total, 228 points have been sampled, at least in one occasion, in all types of water bodies (rivers, canals, lagoons, marshes). In 106 points, more than two sampling events have been performed. Furthermore, in 2025, a specific diagnosis was carried out to better understand the current situation of the species in these protected spaces after a prolonged and severe drought. Sampling was carried out everywhere with fykenets, adding the use of nets and electrofishing in some stations located in large bodies of water. The known historical evolution of Spanish toothcarp distribution in the Empordà has been presented since the first data was made available at the beginning of the 1980s of the 20th century until now. Today, this fish only occupies approximately 10% of its potential distribution in the area. Over the last five decades, its distribution has expanded and contracted several times, but in 2025, it was once again in the worst possible situation of the analysis period. Therefore, the implementation of protected areas, or the execution of several large ecological restoration projects, have only succeeded in maintaining existing populations, but not in the expansion of its potential distribution to unoccupied areas. The main factors that explain this general situation are mainly (1) the proliferation of exotic species, (2) the loss of ecological status, and (3) hydrological alterations and increasingly severe droughts. Thus, the recovery and long-term conservation of Spanish toothcarp in the Empordà area inevitably requires the implementation of more extensive and decisive measures to reverse the incidence of these factors.

]]> Diagnosis of the Present Situation of the Spanish Toothcarp (Apricaphanius iberus) in Empordà Wetlands Quim Pou-Rovira Neus Mairal Elena Farré Bernat Dalmau Llorenç Ferrer Maria Spotti Eloi Cruset doi: 10.3390/proceedings2026146109 Proceedings 2026-06-22 Proceedings 2026-06-22 146 1 Abstract 109 10.3390/proceedings2026146109 https://www.mdpi.com/2504-3900/146/1/109 Proceedings, Vol. 146, Pages 106: Landmark-Based Geometric Morphometrics Approach to Unravel the Population Structure of the Bluefish (Pomatomus saltatrix) in Southeastern–South Brazil https://www.mdpi.com/2504-3900/146/1/106 Introduction: Understanding population structure is essential for the effective management of marine resources. The bluefish (Pomatomus saltatrix) is a widely distributed and commercially important species along the Brazilian coast, yet its population structure in the South–Southeastern region remains unclear. Objectives: This study aimed to assess the body morphological variation and to infer the population structure of P. saltatrix along the South and Southeastern coasts of Brazil using a landmark-based geometric morphometrics method. Methodology: Individuals were collected between August and October 2024 from artisanal fisheries in Rio de Janeiro (n = 46), São Paulo (n = 37), and Santa Catarina (n = 29). A total of 12 landmarks were used to derive a truss network and extract 25 transformed distances (DTs). Data were analyzed using univariate (one-way ANOVA, followed by Tukey’s test if needed, or Kruskal–Wallis followed by Dunn’s test if needed) and multivariate statistics (PERMANOVA and Flexible Discriminant Analysis (FDA)). Results: Significant regional differences were detected for most DTs, except DT3, DT4, DT6, DT9, DT13, DT24, and DT25. PERMANOVA revealed significant differences among all sampling locations, including pairwise comparisons (p < 0.05). FDA achieved an overall reclassification success of 93% of individuals to their original location. Conclusion: Despite the high reclassification accuracy, the results support the existence of a single stock with spatial structuring rather than completely distinct population units, indicating that P. saltatrix in this region is not homogeneous. 2026-06-22 Proceedings, Vol. 146, Pages 106: Landmark-Based Geometric Morphometrics Approach to Unravel the Population Structure of the Bluefish (Pomatomus saltatrix) in Southeastern–South Brazil

Proceedings doi: 10.3390/proceedings2026146106

Authors: Bianca Neves Felippe Alexandre Daros Rafael Schroeder Rafael Gaio Kulzer Marcus Rodrigues da Costa Rodolfo Miguel Silva Alberto Teodorico Correia

Introduction: Understanding population structure is essential for the effective management of marine resources. The bluefish (Pomatomus saltatrix) is a widely distributed and commercially important species along the Brazilian coast, yet its population structure in the South–Southeastern region remains unclear. Objectives: This study aimed to assess the body morphological variation and to infer the population structure of P. saltatrix along the South and Southeastern coasts of Brazil using a landmark-based geometric morphometrics method. Methodology: Individuals were collected between August and October 2024 from artisanal fisheries in Rio de Janeiro (n = 46), São Paulo (n = 37), and Santa Catarina (n = 29). A total of 12 landmarks were used to derive a truss network and extract 25 transformed distances (DTs). Data were analyzed using univariate (one-way ANOVA, followed by Tukey’s test if needed, or Kruskal–Wallis followed by Dunn’s test if needed) and multivariate statistics (PERMANOVA and Flexible Discriminant Analysis (FDA)). Results: Significant regional differences were detected for most DTs, except DT3, DT4, DT6, DT9, DT13, DT24, and DT25. PERMANOVA revealed significant differences among all sampling locations, including pairwise comparisons (p < 0.05). FDA achieved an overall reclassification success of 93% of individuals to their original location. Conclusion: Despite the high reclassification accuracy, the results support the existence of a single stock with spatial structuring rather than completely distinct population units, indicating that P. saltatrix in this region is not homogeneous.

]]> Landmark-Based Geometric Morphometrics Approach to Unravel the Population Structure of the Bluefish (Pomatomus saltatrix) in Southeastern–South Brazil Bianca Neves Felippe Alexandre Daros Rafael Schroeder Rafael Gaio Kulzer Marcus Rodrigues da Costa Rodolfo Miguel Silva Alberto Teodorico Correia doi: 10.3390/proceedings2026146106 Proceedings 2026-06-22 Proceedings 2026-06-22 146 1 Abstract 106 10.3390/proceedings2026146106 https://www.mdpi.com/2504-3900/146/1/106 Proceedings, Vol. 146, Pages 105: Image Analysis Criteria for the Macroscopic Assessment of Skin Healing in Atlantic Salmon https://www.mdpi.com/2504-3900/146/1/105 Introduction: Fish skin is the first line of defense against the aquatic environment, acting as a physical, chemical, and immunological barrier. In addition to preventing pathogen entry, the skin and its mucus contribute to osmoregulation, innate immunity, and redox balance. Skin lesions—caused by mechanical damage, parasites, environmental stress, or handling—disrupt this barrier, increasing susceptibility to infections, inflammation, and production losses. Thus, efficient skin regeneration is essential for fish welfare and performance. Nutrition plays a key role in this process by providing substrates for epithelial repair, immune function, and antioxidant defense. Among dietary factors, zinc (Zn) is particularly important due to its involvement in cell proliferation, enzymatic activity, and maintenance of skin integrity. Objective: Our objective is to assess the effectiveness of image-based analysis in quantifying the skin healing process in Atlantic salmon fed diets supplemented with zinc. Methodology: The trial comprised three dietary treatments: a control diet with 42 mg Zn per kg (D1), and two diets supplemented up to 120 mg/kg of zinc, derived from inorganic (D2) or organic (D3) forms. Pit-tagged fish with an initial body weight (78 ± 0.1 g) were fed the diets for 75 days. After 15 days of experimental feeding, a standardized wound lesion (2.5 mm diameter × 0.5 mm depth) was inflicted in deeply anesthetized fish, with a disposable biopsy punch, in the dorsal area. After wound infliction, the fish resumed their normal feeding regime for the rest of the trial days. The progression of skin wound healing was assessed using standardized digital image analysis. High-resolution photographs of individual wounds were collected 8, 16, 24 and 32 days post-wounding. All images were acquired under standardized conditions with the inclusion of ArUco identifiers to enable a subsequent computer-assisted comparison. Morphometric parameters (wound width, diameter, perimeter and area) were used to assess wound contraction and closure over time. In parallel, a semi-quantitative visual scoring system was applied to each wound image to capture qualitative aspects of healing that are not fully described by morphometric data alone. Results: Full data analysis is currently underway, but the first results show beneficial effects of dietary zinc supplementation on the skin regenerative process. Conclusions: The combined use of objective digital measurements and standardized visual scoring enabled a comprehensive evaluation of wound healing progress, bridging quantitative tissue remodeling with biologically relevant phenotypic outcomes. This image-based framework provides a sensitive and reproducible approach for assessing dietary interventions targeting skin regeneration and barrier restoration in Atlantic salmon. 2026-06-22 Proceedings, Vol. 146, Pages 105: Image Analysis Criteria for the Macroscopic Assessment of Skin Healing in Atlantic Salmon

Proceedings doi: 10.3390/proceedings2026146105

Authors: João Leça Bruna Henriques Filipe Soares Cláudia Magalhães Rui Rocha Paulo Rema

Introduction: Fish skin is the first line of defense against the aquatic environment, acting as a physical, chemical, and immunological barrier. In addition to preventing pathogen entry, the skin and its mucus contribute to osmoregulation, innate immunity, and redox balance. Skin lesions—caused by mechanical damage, parasites, environmental stress, or handling—disrupt this barrier, increasing susceptibility to infections, inflammation, and production losses. Thus, efficient skin regeneration is essential for fish welfare and performance. Nutrition plays a key role in this process by providing substrates for epithelial repair, immune function, and antioxidant defense. Among dietary factors, zinc (Zn) is particularly important due to its involvement in cell proliferation, enzymatic activity, and maintenance of skin integrity. Objective: Our objective is to assess the effectiveness of image-based analysis in quantifying the skin healing process in Atlantic salmon fed diets supplemented with zinc. Methodology: The trial comprised three dietary treatments: a control diet with 42 mg Zn per kg (D1), and two diets supplemented up to 120 mg/kg of zinc, derived from inorganic (D2) or organic (D3) forms. Pit-tagged fish with an initial body weight (78 ± 0.1 g) were fed the diets for 75 days. After 15 days of experimental feeding, a standardized wound lesion (2.5 mm diameter × 0.5 mm depth) was inflicted in deeply anesthetized fish, with a disposable biopsy punch, in the dorsal area. After wound infliction, the fish resumed their normal feeding regime for the rest of the trial days. The progression of skin wound healing was assessed using standardized digital image analysis. High-resolution photographs of individual wounds were collected 8, 16, 24 and 32 days post-wounding. All images were acquired under standardized conditions with the inclusion of ArUco identifiers to enable a subsequent computer-assisted comparison. Morphometric parameters (wound width, diameter, perimeter and area) were used to assess wound contraction and closure over time. In parallel, a semi-quantitative visual scoring system was applied to each wound image to capture qualitative aspects of healing that are not fully described by morphometric data alone. Results: Full data analysis is currently underway, but the first results show beneficial effects of dietary zinc supplementation on the skin regenerative process. Conclusions: The combined use of objective digital measurements and standardized visual scoring enabled a comprehensive evaluation of wound healing progress, bridging quantitative tissue remodeling with biologically relevant phenotypic outcomes. This image-based framework provides a sensitive and reproducible approach for assessing dietary interventions targeting skin regeneration and barrier restoration in Atlantic salmon.

]]> Image Analysis Criteria for the Macroscopic Assessment of Skin Healing in Atlantic Salmon João Leça Bruna Henriques Filipe Soares Cláudia Magalhães Rui Rocha Paulo Rema doi: 10.3390/proceedings2026146105 Proceedings 2026-06-22 Proceedings 2026-06-22 146 1 Abstract 105 10.3390/proceedings2026146105 https://www.mdpi.com/2504-3900/146/1/105 Proceedings, Vol. 146, Pages 99: Genomic and Phylogenetic Insights into the Hybridogenetic Origin of the Probably Extinct Iberian Endemic Squalius palaciosi https://www.mdpi.com/2504-3900/146/1/99 Introduction: Squalius palaciosi (Doadrio, 1980; Leuciscidae) is a highly threatened freshwater fish species with an extremely restricted distribution, currently confined to a few tributaries on the right bank of the Guadalquivir River basin. During the 1980s, its populations were abundant and constituted a dominant component of local fish communities. However, multiple threats led to a drastic population decline, bringing the species to the brink of extinction. From an evolutionary perspective, S. palaciosi is particularly remarkable due to its polyploid condition and its potential involvement in hybridogenetic complexes, a rare phenomenon in the Iberian Peninsula. Hybridogenetic systems are well documented in its congeners Squalius alburnoides, widely distributed across Iberian river basins, and Squalius sp., restricted to the Guadiana basin. In these systems, the maternal lineage is shared (Squalius pyrenaicus), whereas the paternal lineage varies and remains unknown in S. palaciosi. Objective: This study aims to generate the first genomic data for S. palaciosi and to elucidate its evolutionary origin, as well as its mitochondrial and nuclear phylogenetic relationships within hybridogenetic complexes. Methodology: Genomic DNA was extracted from skeletal remains of preserved specimens housed in the fish collection of the National Museum of Natural Sciences (MNCN-CSIC) and subjected to Illumina short-read sequencing. After quality filtering, potential contaminant reads were removed. The complete mitochondrial genome and several nuclear gene fragments were assembled. Mitochondrial phylogenetic analyses were conducted using publicly available whole-genome sequencing data from Iberian freshwater fish species. Nuclear gene fragments were taxonomically assigned using BLAST analyses. Results: Phylogenetic analyses revealed that S. palaciosi is closely related at the mitochondrial level to S. alburnoides and S. tartessicus, with strong statistical support. BLAST-based taxonomic assignments of nuclear markers suggest the involvement of multiple Iberian freshwater fish species in the hybridogenetic origin of S. palaciosi. Conclusions: Our results provide novel insights into the evolutionary history of S. palaciosi and support a complex hybridogenetic origin involving multiple parental lineages. This study contributes to a better understanding of hybridogenetic speciation in freshwater fishes, a rare but evolutionarily significant process. 2026-06-22 Proceedings, Vol. 146, Pages 99: Genomic and Phylogenetic Insights into the Hybridogenetic Origin of the Probably Extinct Iberian Endemic Squalius palaciosi

Proceedings doi: 10.3390/proceedings2026146099

Authors: Silvia Perea Miriam Casal-López Hamid Reza Ghanavi Ignacio Doadrio

Introduction: Squalius palaciosi (Doadrio, 1980; Leuciscidae) is a highly threatened freshwater fish species with an extremely restricted distribution, currently confined to a few tributaries on the right bank of the Guadalquivir River basin. During the 1980s, its populations were abundant and constituted a dominant component of local fish communities. However, multiple threats led to a drastic population decline, bringing the species to the brink of extinction. From an evolutionary perspective, S. palaciosi is particularly remarkable due to its polyploid condition and its potential involvement in hybridogenetic complexes, a rare phenomenon in the Iberian Peninsula. Hybridogenetic systems are well documented in its congeners Squalius alburnoides, widely distributed across Iberian river basins, and Squalius sp., restricted to the Guadiana basin. In these systems, the maternal lineage is shared (Squalius pyrenaicus), whereas the paternal lineage varies and remains unknown in S. palaciosi. Objective: This study aims to generate the first genomic data for S. palaciosi and to elucidate its evolutionary origin, as well as its mitochondrial and nuclear phylogenetic relationships within hybridogenetic complexes. Methodology: Genomic DNA was extracted from skeletal remains of preserved specimens housed in the fish collection of the National Museum of Natural Sciences (MNCN-CSIC) and subjected to Illumina short-read sequencing. After quality filtering, potential contaminant reads were removed. The complete mitochondrial genome and several nuclear gene fragments were assembled. Mitochondrial phylogenetic analyses were conducted using publicly available whole-genome sequencing data from Iberian freshwater fish species. Nuclear gene fragments were taxonomically assigned using BLAST analyses. Results: Phylogenetic analyses revealed that S. palaciosi is closely related at the mitochondrial level to S. alburnoides and S. tartessicus, with strong statistical support. BLAST-based taxonomic assignments of nuclear markers suggest the involvement of multiple Iberian freshwater fish species in the hybridogenetic origin of S. palaciosi. Conclusions: Our results provide novel insights into the evolutionary history of S. palaciosi and support a complex hybridogenetic origin involving multiple parental lineages. This study contributes to a better understanding of hybridogenetic speciation in freshwater fishes, a rare but evolutionarily significant process.

]]> Genomic and Phylogenetic Insights into the Hybridogenetic Origin of the Probably Extinct Iberian Endemic Squalius palaciosi Silvia Perea Miriam Casal-López Hamid Reza Ghanavi Ignacio Doadrio doi: 10.3390/proceedings2026146099 Proceedings 2026-06-22 Proceedings 2026-06-22 146 1 Abstract 99 10.3390/proceedings2026146099 https://www.mdpi.com/2504-3900/146/1/99 Proceedings, Vol. 146, Pages 102: Freshwater Aquarium Fish Imports: From Species and Quantities to Origins and Risks https://www.mdpi.com/2504-3900/146/1/102 Introduction: The global ornamental fish trade is a rapidly expanding sector and a major pathway for the introduction of non-native species, particularly in freshwater ecosystems in developed countries. The introduction of non-native species can result in a range of ecological impacts, including predation, competition, hybridization, and disease transmission, often leading to ecosystem degradation and biotic homogenization. Therefore, it represents a clear ecological risk, especially serious in freshwater systems with a high endemism rate, such as the Iberian Peninsula. The occurrence of ornamental non-native species in the Iberian Peninsula has been common, yet little has been done to describe the overall ornamental fish trade as a first step to evaluate invasion risk. Objective: This study characterizes the import dynamics of ornamental freshwater fish in Portugal between 2020 and 2024 and evaluates its potential role as a pathway for species introductions. Methodology: Data were obtained from the Institute for Nature Conservation and Forests database, including information on species composition, quantities, sizes, prices, and countries of origin. A total of 431 records were analyzed, resulting in 27,689 validated entries of imported freshwater fish, which were taxonomically verified and filtered to retain only freshwater species. Results: A total of 666 species from 88 families were identified, with an average of 380 species imported annually, reflecting high taxonomic diversity. Import volumes increased from approximately 1.25 million individuals in 2020 to 1.75 million in 2024, while total import value nearly doubled from €300,000 to €600,000. Imports were predominantly from five Southeast Asian countries, particularly Indonesia and Vietnam, and largely supported by aquaculture production (88%). A stable core of highly traded species, including Carassius auratus, Poecilia reticulata, and Paracheirodon innesi, suggests a sustained and very high propagule pressure, while some species variability was observed on yearly basis, suggesting the importance of monitoring programs on actual imports. Conclusions: Overall, the ornamental fish trade represents a significant and growing pathway for biological invasions in Portugal. The combination of increasing trade volume, high species diversity, and persistent dominance of key taxa highlights the need for improved monitoring, regulatory frameworks, and public awareness to mitigate ecological risks. 2026-06-22 Proceedings, Vol. 146, Pages 102: Freshwater Aquarium Fish Imports: From Species and Quantities to Origins and Risks

Proceedings doi: 10.3390/proceedings2026146102

Authors: Luísa Sousa Carla Silva Pedro Anastácio Filipe Ribeiro

Introduction: The global ornamental fish trade is a rapidly expanding sector and a major pathway for the introduction of non-native species, particularly in freshwater ecosystems in developed countries. The introduction of non-native species can result in a range of ecological impacts, including predation, competition, hybridization, and disease transmission, often leading to ecosystem degradation and biotic homogenization. Therefore, it represents a clear ecological risk, especially serious in freshwater systems with a high endemism rate, such as the Iberian Peninsula. The occurrence of ornamental non-native species in the Iberian Peninsula has been common, yet little has been done to describe the overall ornamental fish trade as a first step to evaluate invasion risk. Objective: This study characterizes the import dynamics of ornamental freshwater fish in Portugal between 2020 and 2024 and evaluates its potential role as a pathway for species introductions. Methodology: Data were obtained from the Institute for Nature Conservation and Forests database, including information on species composition, quantities, sizes, prices, and countries of origin. A total of 431 records were analyzed, resulting in 27,689 validated entries of imported freshwater fish, which were taxonomically verified and filtered to retain only freshwater species. Results: A total of 666 species from 88 families were identified, with an average of 380 species imported annually, reflecting high taxonomic diversity. Import volumes increased from approximately 1.25 million individuals in 2020 to 1.75 million in 2024, while total import value nearly doubled from €300,000 to €600,000. Imports were predominantly from five Southeast Asian countries, particularly Indonesia and Vietnam, and largely supported by aquaculture production (88%). A stable core of highly traded species, including Carassius auratus, Poecilia reticulata, and Paracheirodon innesi, suggests a sustained and very high propagule pressure, while some species variability was observed on yearly basis, suggesting the importance of monitoring programs on actual imports. Conclusions: Overall, the ornamental fish trade represents a significant and growing pathway for biological invasions in Portugal. The combination of increasing trade volume, high species diversity, and persistent dominance of key taxa highlights the need for improved monitoring, regulatory frameworks, and public awareness to mitigate ecological risks.

]]> Freshwater Aquarium Fish Imports: From Species and Quantities to Origins and Risks Luísa Sousa Carla Silva Pedro Anastácio Filipe Ribeiro doi: 10.3390/proceedings2026146102 Proceedings 2026-06-22 Proceedings 2026-06-22 146 1 Abstract 102 10.3390/proceedings2026146102 https://www.mdpi.com/2504-3900/146/1/102 Proceedings, Vol. 146, Pages 107: Multisubstance Screening Supports a High-Throughput Zebrafish Thigmotaxis Assay for One Health-Oriented Neurotoxicity Assessment https://www.mdpi.com/2504-3900/146/1/107 Introduction: Aquatic contaminants can alter fish behavior before overt toxicity becomes evident, making neurobehavioral endpoints relevant for ecosystem protection and for hazard prioritization within a One Health framework. We recently developed a high-throughput visual-acoustic zebrafish larval thigmotaxis assay in which edge preference is interpreted as an anxiety-like behavioral endpoint, thereby adding spatial phenotyping beyond conventional locomotion metrics. Objective: To evaluate assay performance in a multisubstance screening challenge and determine whether it can discriminate distinct behavioral fingerprints without prior knowledge of chemical identity. Methodology: Zebrafish larvae were exposed for 1 h at 120 hpf. For each substance, 24 larvae were tested per condition, with six concentrations per substance, plus positive and negative controls. Larvae were challenged using alternating light/dark and tapping/quiet paradigms. The primary endpoint was the percentage of time spent at the edge as a proxy for anxiety-like behavior, while total distance and mean total velocity when moving were used as contextual locomotor metrics; edge distance and edge velocity were used as supportive spatial metrics. Data from 37 substances were analyzed through a standardized automated workflow. Results: Controls performed as expected and supported assay stability across runs. The chemical screening revealed heterogeneous but reproducible behavioral fingerprints. Seven substances produced weak/minimal acute responses, ten showed predominantly suppressive profiles, three predominantly activating profiles, nine showed prominent thigmotaxis-specific anxiety-like signals not explained by locomotion alone, and eight displayed mixed or stimulus-dependent patterns, including non-monotonic responses. Several substances altered edge preference while distance and velocity changed less, differently, or in the opposite direction, indicating behavioral reorganization rather than simple hypo- or hyperactivity. The multi-stimulus design was critical because some effects were evident only under specific sensory contexts. Conclusions: The multisubstance challenge supports the discriminatory capacity, robustness, and added value of the assay for high-throughput neurobehavioral screening. By capturing anxiety-like behavior through thigmotaxis and complementing it with locomotor context, the method improves phenotypic resolution for aquatic pollution assessment and offers a sensitive fish-based NAM to prioritize chemicals of concern for both environmental and human health-oriented testing strategies. 2026-06-22 Proceedings, Vol. 146, Pages 107: Multisubstance Screening Supports a High-Throughput Zebrafish Thigmotaxis Assay for One Health-Oriented Neurotoxicity Assessment

Proceedings doi: 10.3390/proceedings2026146107

Authors: Monica Torres-Ruiz María Muñoz-Palencia Laura Sánchez-Ramos Ana I. Cañas-Portilla Antonio de la Vieja

Introduction: Aquatic contaminants can alter fish behavior before overt toxicity becomes evident, making neurobehavioral endpoints relevant for ecosystem protection and for hazard prioritization within a One Health framework. We recently developed a high-throughput visual-acoustic zebrafish larval thigmotaxis assay in which edge preference is interpreted as an anxiety-like behavioral endpoint, thereby adding spatial phenotyping beyond conventional locomotion metrics. Objective: To evaluate assay performance in a multisubstance screening challenge and determine whether it can discriminate distinct behavioral fingerprints without prior knowledge of chemical identity. Methodology: Zebrafish larvae were exposed for 1 h at 120 hpf. For each substance, 24 larvae were tested per condition, with six concentrations per substance, plus positive and negative controls. Larvae were challenged using alternating light/dark and tapping/quiet paradigms. The primary endpoint was the percentage of time spent at the edge as a proxy for anxiety-like behavior, while total distance and mean total velocity when moving were used as contextual locomotor metrics; edge distance and edge velocity were used as supportive spatial metrics. Data from 37 substances were analyzed through a standardized automated workflow. Results: Controls performed as expected and supported assay stability across runs. The chemical screening revealed heterogeneous but reproducible behavioral fingerprints. Seven substances produced weak/minimal acute responses, ten showed predominantly suppressive profiles, three predominantly activating profiles, nine showed prominent thigmotaxis-specific anxiety-like signals not explained by locomotion alone, and eight displayed mixed or stimulus-dependent patterns, including non-monotonic responses. Several substances altered edge preference while distance and velocity changed less, differently, or in the opposite direction, indicating behavioral reorganization rather than simple hypo- or hyperactivity. The multi-stimulus design was critical because some effects were evident only under specific sensory contexts. Conclusions: The multisubstance challenge supports the discriminatory capacity, robustness, and added value of the assay for high-throughput neurobehavioral screening. By capturing anxiety-like behavior through thigmotaxis and complementing it with locomotor context, the method improves phenotypic resolution for aquatic pollution assessment and offers a sensitive fish-based NAM to prioritize chemicals of concern for both environmental and human health-oriented testing strategies.

]]> Multisubstance Screening Supports a High-Throughput Zebrafish Thigmotaxis Assay for One Health-Oriented Neurotoxicity Assessment Monica Torres-Ruiz María Muñoz-Palencia Laura Sánchez-Ramos Ana I. Cañas-Portilla Antonio de la Vieja doi: 10.3390/proceedings2026146107 Proceedings 2026-06-22 Proceedings 2026-06-22 146 1 Abstract 107 10.3390/proceedings2026146107 https://www.mdpi.com/2504-3900/146/1/107 Proceedings, Vol. 146, Pages 98: Heat Hardening in Grey Mullets: Physiological Responses of Juvenile Chelon labrosus and Chelon aurata Under Simulated Short-Term Marine Heatwaves https://www.mdpi.com/2504-3900/146/1/98 Introduction: Marine heatwaves are increasing in frequency and intensity, posing major challenges for fishes inhabiting shallow coastal ecosystems. Short-term exposure to extreme warming can alter metabolic performance and thermal tolerance, with potential consequences for species persistence and school composition in thermally variable habitats. Understanding the capacity of coastal fishes to withstand acute warming events is therefore essential for predicting ecological responses to climate change. Objective: We aimed to determine the effects of simulated marine heatwaves on thermal tolerance and metabolic performance in juvenile grey mullets, Chelon labrosus and Chelon aurata, two abundant sympatric species inhabiting the Ria Formosa lagoon (southern Portugal). Methodology: Juvenile mullets acclimated at 17 °C were exposed to simulated heatwave treatments of 23, 27, or 33 °C and sampled either at peak temperature or after 48 h and 1-week recovery at 17 °C. Critical thermal maximum (CTmax, using a 1 °C/min thermal ramp), static oxygen consumption (MO2), and intermittent respirometry parameters were measured. Standard metabolic rate (SMR), maximum metabolic rate (MMR), and aerobic scope (AS) were derived from intermittent respirometry. A complementary temperature-ramp (>3 h at each temperature step 17, 23, 27 and 33 °C) was performed to evaluate routine metabolic rate and estimate Q10 values across increasing temperatures. Additional plasma and tissue analyses are being conducted to assess energetic substrate mobilization and cellular responses to thermal and oxidative stress. Results: CTmax increased significantly with warming in both treatment modes, demonstrating rapid heat hardening in juvenile mullets. Fish exposed to 27 and 33 °C exhibited higher CTmax than control fish, and this elevated tolerance persisted after recovery. Chelon labrosus showed slightly higher CTmax values than C. aurata. Oxygen consumption increased with temperature, with the strongest responses occurring at 33 °C. SMR increased markedly with warming, particularly in heatwave-exposed fish, while MMR increased mainly at the highest temperature treatment. In contrast, AS showed no clear thermal optimum or decline across treatments. Routine metabolic rate increased non-linearly with temperature in the complementary ramp experiment, with a mean Q10 of 2.28, confirming strong thermal dependence of metabolism. Conclusions: Juvenile mullets possess substantial short-term thermal plasticity and can rapidly increase heat tolerance during marine heatwaves but this enhanced tolerance is accompanied by elevated metabolic costs under extreme warming, indicating potential energetic trade-offs near upper thermal limits. Differential physiological responses between species may influence school composition and ecological performance across thermal landscapes. Ongoing plasma and tissue analyses will further clarify the energetic and cellular mechanisms underlying thermal and oxidative stress resilience in coastal fishes. 2026-06-22 Proceedings, Vol. 146, Pages 98: Heat Hardening in Grey Mullets: Physiological Responses of Juvenile Chelon labrosus and Chelon aurata Under Simulated Short-Term Marine Heatwaves

Proceedings doi: 10.3390/proceedings2026146098

Authors: Inês Amaral Rita A. Costa Antonio Zamora-López Wim Zimmermann Adrián Guerrero-Gómez Sílvia F. Gregório Pedro M. Guerreiro

Introduction: Marine heatwaves are increasing in frequency and intensity, posing major challenges for fishes inhabiting shallow coastal ecosystems. Short-term exposure to extreme warming can alter metabolic performance and thermal tolerance, with potential consequences for species persistence and school composition in thermally variable habitats. Understanding the capacity of coastal fishes to withstand acute warming events is therefore essential for predicting ecological responses to climate change. Objective: We aimed to determine the effects of simulated marine heatwaves on thermal tolerance and metabolic performance in juvenile grey mullets, Chelon labrosus and Chelon aurata, two abundant sympatric species inhabiting the Ria Formosa lagoon (southern Portugal). Methodology: Juvenile mullets acclimated at 17 °C were exposed to simulated heatwave treatments of 23, 27, or 33 °C and sampled either at peak temperature or after 48 h and 1-week recovery at 17 °C. Critical thermal maximum (CTmax, using a 1 °C/min thermal ramp), static oxygen consumption (MO2), and intermittent respirometry parameters were measured. Standard metabolic rate (SMR), maximum metabolic rate (MMR), and aerobic scope (AS) were derived from intermittent respirometry. A complementary temperature-ramp (>3 h at each temperature step 17, 23, 27 and 33 °C) was performed to evaluate routine metabolic rate and estimate Q10 values across increasing temperatures. Additional plasma and tissue analyses are being conducted to assess energetic substrate mobilization and cellular responses to thermal and oxidative stress. Results: CTmax increased significantly with warming in both treatment modes, demonstrating rapid heat hardening in juvenile mullets. Fish exposed to 27 and 33 °C exhibited higher CTmax than control fish, and this elevated tolerance persisted after recovery. Chelon labrosus showed slightly higher CTmax values than C. aurata. Oxygen consumption increased with temperature, with the strongest responses occurring at 33 °C. SMR increased markedly with warming, particularly in heatwave-exposed fish, while MMR increased mainly at the highest temperature treatment. In contrast, AS showed no clear thermal optimum or decline across treatments. Routine metabolic rate increased non-linearly with temperature in the complementary ramp experiment, with a mean Q10 of 2.28, confirming strong thermal dependence of metabolism. Conclusions: Juvenile mullets possess substantial short-term thermal plasticity and can rapidly increase heat tolerance during marine heatwaves but this enhanced tolerance is accompanied by elevated metabolic costs under extreme warming, indicating potential energetic trade-offs near upper thermal limits. Differential physiological responses between species may influence school composition and ecological performance across thermal landscapes. Ongoing plasma and tissue analyses will further clarify the energetic and cellular mechanisms underlying thermal and oxidative stress resilience in coastal fishes.

]]> Heat Hardening in Grey Mullets: Physiological Responses of Juvenile Chelon labrosus and Chelon aurata Under Simulated Short-Term Marine Heatwaves Inês Amaral Rita A. Costa Antonio Zamora-López Wim Zimmermann Adrián Guerrero-Gómez Sílvia F. Gregório Pedro M. Guerreiro doi: 10.3390/proceedings2026146098 Proceedings 2026-06-22 Proceedings 2026-06-22 146 1 Abstract 98 10.3390/proceedings2026146098 https://www.mdpi.com/2504-3900/146/1/98 Proceedings, Vol. 146, Pages 103: Range Expansion and Shifting Aggregation Dynamics of Tub Gurnard Trigla lyra (Scorpaenoidei: Triglidae) in Galician and Cantabrian Sea Waters (NE Atlantic, 1993–2025) https://www.mdpi.com/2504-3900/146/1/103 Introduction: The tub gurnard (Trigla lyra) is a demersal species of commercial interest whose long-term distributional dynamics remain poorly understood. Understanding spatial and temporal changes is essential for fisheries management and for assessing biogeographic shifts. Objective: To characterise the spatio-temporal distribution and persistence of T. lyra across Galician and Cantabrian Sea waters over a 33-year period (1993–2025) and to identify environmental and fishing drivers associated with observed changes. Methodology: We analysed data from the DEMERSALES bottom trawl survey series (1993–2025), for which the sampling design remained consistent throughout. Species distribution was modelled using a delta–GAM framework (presence–absence and positive values), complemented by a presence-only GAM fitted to Vessel Monitoring System (VMS) data; because these data were only available for 2009–2023, this model was restricted to that period for biological coherence. Environmental predictors included bathymetry, slope, sediment composition (organic matter, mud, fine and coarse sand), bottom temperature, and salinity. Spatial structure was assessed using aggregation curves, occupied area, centre of gravity, a Space Selectivity Index, and an Index of Persistence. Results: The occupied area increased from 45 to 963 km2 (+2040%), accompanied by a sustained decline in the Space Selectivity Index and a westward shift of the distributional centroid (~20 km), indicating progressive range broadening. The frequency of occurrence rose from 4.5% in 1993 to 87.7% in 2025, reflecting a marked increase in spatial occupancy and encounter probability. Abundance increased sharply after 2015 (+47%), consistent with strong positive year effects in the GAM. Higher occurrence and densities were associated with muddy substrates, intermediate to high organic content, and depths of 100–300 m, matching the stable aggregation cores found along the shelf break. A reduction in trawling effort (−38% in mean intensity, −17% in swept area over 14 years) likely facilitated these trends. Conclusions: T. lyra expanded its distribution and shifted westward between 1993 and 2025, with persistent aggregation cores on the shelf break. No significant effect of temperature was found, suggesting that climate warming is not the primary driver; the expansion appears most plausibly to have been favoured by the decline in fishing pressure. 2026-06-22 Proceedings, Vol. 146, Pages 103: Range Expansion and Shifting Aggregation Dynamics of Tub Gurnard Trigla lyra (Scorpaenoidei: Triglidae) in Galician and Cantabrian Sea Waters (NE Atlantic, 1993–2025)

Proceedings doi: 10.3390/proceedings2026146103

Authors: Juan Carlos Arronte Jose Manuel González-Irusta Francisco Velasco Alberto Serrano

Introduction: The tub gurnard (Trigla lyra) is a demersal species of commercial interest whose long-term distributional dynamics remain poorly understood. Understanding spatial and temporal changes is essential for fisheries management and for assessing biogeographic shifts. Objective: To characterise the spatio-temporal distribution and persistence of T. lyra across Galician and Cantabrian Sea waters over a 33-year period (1993–2025) and to identify environmental and fishing drivers associated with observed changes. Methodology: We analysed data from the DEMERSALES bottom trawl survey series (1993–2025), for which the sampling design remained consistent throughout. Species distribution was modelled using a delta–GAM framework (presence–absence and positive values), complemented by a presence-only GAM fitted to Vessel Monitoring System (VMS) data; because these data were only available for 2009–2023, this model was restricted to that period for biological coherence. Environmental predictors included bathymetry, slope, sediment composition (organic matter, mud, fine and coarse sand), bottom temperature, and salinity. Spatial structure was assessed using aggregation curves, occupied area, centre of gravity, a Space Selectivity Index, and an Index of Persistence. Results: The occupied area increased from 45 to 963 km2 (+2040%), accompanied by a sustained decline in the Space Selectivity Index and a westward shift of the distributional centroid (~20 km), indicating progressive range broadening. The frequency of occurrence rose from 4.5% in 1993 to 87.7% in 2025, reflecting a marked increase in spatial occupancy and encounter probability. Abundance increased sharply after 2015 (+47%), consistent with strong positive year effects in the GAM. Higher occurrence and densities were associated with muddy substrates, intermediate to high organic content, and depths of 100–300 m, matching the stable aggregation cores found along the shelf break. A reduction in trawling effort (−38% in mean intensity, −17% in swept area over 14 years) likely facilitated these trends. Conclusions: T. lyra expanded its distribution and shifted westward between 1993 and 2025, with persistent aggregation cores on the shelf break. No significant effect of temperature was found, suggesting that climate warming is not the primary driver; the expansion appears most plausibly to have been favoured by the decline in fishing pressure.

]]> Range Expansion and Shifting Aggregation Dynamics of Tub Gurnard Trigla lyra (Scorpaenoidei: Triglidae) in Galician and Cantabrian Sea Waters (NE Atlantic, 1993–2025) Juan Carlos Arronte Jose Manuel González-Irusta Francisco Velasco Alberto Serrano doi: 10.3390/proceedings2026146103 Proceedings 2026-06-22 Proceedings 2026-06-22 146 1 Abstract 103 10.3390/proceedings2026146103 https://www.mdpi.com/2504-3900/146/1/103 Proceedings, Vol. 146, Pages 91: Population Stability of the Endangered Iberian Killifish in Coastal Salt Pans (SE Spain): Hydrological Management as the Main Drive https://www.mdpi.com/2504-3900/146/1/91 Introduction: The Iberian killifish (Apricaphanius iberus), an endemic cyprinodontid of the Spanish Mediterranean coast, is a key conservation species due to its restricted distribution, high sensitivity to habitat alteration and invasive fish, and ecological role in saline wetlands. Classified as Endangered by national administration and listed in Annex II of the EU Habitats Directive, its populations have undergone severe declines across its native range. Coastal salt pans have become critical refuges for the persistence of this species, often supporting some of the highest population densities recorded. However, the conservation value of these anthropogenic systems strongly depends on hydrological management, which directly influences habitat stability and suitability. Objectives: This study aims to assess the long-term population responses of A. iberus under contrasting hydrological management regimes in coastal salt pans. Methodology: We analysed ten years of monitoring data across five salt pan circuits in the perimeter area of the Mar Menor coastal lagoon (SE Spain), covering a salinity gradient and three management regimes: economic exploitation (EE), preventive management (PM), and non-management (NM). Population metrics (occurrence, abundance and breeder proportion) were evaluated. Results: Population performance differed markedly among management regimes. EE circuits showed consistently high occurrence and abundance, including in hypersaline ponds, high breeder proportions, low interannual variability, and weak dependence on meteorological conditions and ephemeral prey. PM circuits displayed intermediate patterns. In contrast, NM circuits exhibited low occurrence and abundance, low breeder proportions, high temporal fluctuations, and strong dependence on meteorological variability. Conclusions: Hydrological management is a key determinant of A. iberus population stability in coastal salt pans. Maintaining or restoring traditional management practices enhances habitat suitability, buffers environmental variability, and supports conservation actions such as reintroduction. These findings highlight the critical role of managed anthropogenic wetlands as refuges for endangered fishes and provide a transferable framework for conservation under global change. 2026-06-22 Proceedings, Vol. 146, Pages 91: Population Stability of the Endangered Iberian Killifish in Coastal Salt Pans (SE Spain): Hydrological Management as the Main Drive

Proceedings doi: 10.3390/proceedings2026146091

Authors: Víctor Manuel Álvarez-Navarro Adrián Guerrero-Gómez Antonio Zamora-López Antonio Andrés Herrero-Reyes Elena Parra-Espín Mar Torralva José Manuel Zamora-Marín José Manuel Vidal-Gil Francisco José Oliva-Paterna

Introduction: The Iberian killifish (Apricaphanius iberus), an endemic cyprinodontid of the Spanish Mediterranean coast, is a key conservation species due to its restricted distribution, high sensitivity to habitat alteration and invasive fish, and ecological role in saline wetlands. Classified as Endangered by national administration and listed in Annex II of the EU Habitats Directive, its populations have undergone severe declines across its native range. Coastal salt pans have become critical refuges for the persistence of this species, often supporting some of the highest population densities recorded. However, the conservation value of these anthropogenic systems strongly depends on hydrological management, which directly influences habitat stability and suitability. Objectives: This study aims to assess the long-term population responses of A. iberus under contrasting hydrological management regimes in coastal salt pans. Methodology: We analysed ten years of monitoring data across five salt pan circuits in the perimeter area of the Mar Menor coastal lagoon (SE Spain), covering a salinity gradient and three management regimes: economic exploitation (EE), preventive management (PM), and non-management (NM). Population metrics (occurrence, abundance and breeder proportion) were evaluated. Results: Population performance differed markedly among management regimes. EE circuits showed consistently high occurrence and abundance, including in hypersaline ponds, high breeder proportions, low interannual variability, and weak dependence on meteorological conditions and ephemeral prey. PM circuits displayed intermediate patterns. In contrast, NM circuits exhibited low occurrence and abundance, low breeder proportions, high temporal fluctuations, and strong dependence on meteorological variability. Conclusions: Hydrological management is a key determinant of A. iberus population stability in coastal salt pans. Maintaining or restoring traditional management practices enhances habitat suitability, buffers environmental variability, and supports conservation actions such as reintroduction. These findings highlight the critical role of managed anthropogenic wetlands as refuges for endangered fishes and provide a transferable framework for conservation under global change.

]]> Population Stability of the Endangered Iberian Killifish in Coastal Salt Pans (SE Spain): Hydrological Management as the Main Drive Víctor Manuel Álvarez-Navarro Adrián Guerrero-Gómez Antonio Zamora-López Antonio Andrés Herrero-Reyes Elena Parra-Espín Mar Torralva José Manuel Zamora-Marín José Manuel Vidal-Gil Francisco José Oliva-Paterna doi: 10.3390/proceedings2026146091 Proceedings 2026-06-22 Proceedings 2026-06-22 146 1 Abstract 91 10.3390/proceedings2026146091 https://www.mdpi.com/2504-3900/146/1/91 Proceedings, Vol. 146, Pages 95: Movement Patterns of the Iberian Barbel (Luciobarbus bocagei Steindachner, 1864) in a Recently Impounded River https://www.mdpi.com/2504-3900/146/1/95 The Iberian barbel (Luciobarbus bocagei Steindachner, 1864) is a potamodromous species that displays migratory movements, with adults moving upstream during spring to spawn, followed by downstream migration in autumn by both adults and juveniles to feed and inhabit more productive river stretches. Increasing river fragmentation due to dam construction and operation causes a loss of connectivity and suitable habitat, which can affect this natural behaviour. In a tributary of River Douro, River Tua, a 108 m high dam, was recently built (i.e., Foz Tua dam in 2017) at only 1.1 km from the river mouth, leaving the upstream spawning area inaccessible. To evaluate the species behavioural response to this impact, a passive acoustic telemetry array was deployed in the study area, and between 2018 and 2023, 120 tagged fish had their movements tracked. The results showed two different behavioural profiles in the same population with migratory (42.5%) and resident (54.0%) individuals. During this period, a specific experimental study was developed to evaluate the response of a subset of 90 fish, captured upstream of the Foz Tua dam and translocated to an alternative downstream Douro tributary (River Pinhão, ~20 km downstream from River Tua). From these, 66.7% remained at the release site, whilst 23.3% returned to the river of origin, i.e., River Tua. From the returned fraction, 28.6% of the tagged fish maintained this migratory behaviour between both rivers in the following years. Generalized Additive Models were used for each of the two behavioural profiles observed in this study, to identify which environmental variables were associated with the presence of the tagged barbels downstream the Foz Tua dam. Ecological flow, temperature and day-period were some of the predictors explaining the use of this river stretch downstream of Foz Tua dam. This study substantially updates the available information regarding this species’ movement patterns at large spatial and temporal scales, contributing to enhancement of management and conservation programs for potamodromous species, in highly impounded and fragmented rivers. 2026-06-22 Proceedings, Vol. 146, Pages 95: Movement Patterns of the Iberian Barbel (Luciobarbus bocagei Steindachner, 1864) in a Recently Impounded River

Proceedings doi: 10.3390/proceedings2026146095

Authors: Ana S. Rato Carlos M. Alexandre Ana F. Silva João P. Marques Sara S. Silva Maria J. Lança Bernardo R. Quintella Pedro R. Almeida

The Iberian barbel (Luciobarbus bocagei Steindachner, 1864) is a potamodromous species that displays migratory movements, with adults moving upstream during spring to spawn, followed by downstream migration in autumn by both adults and juveniles to feed and inhabit more productive river stretches. Increasing river fragmentation due to dam construction and operation causes a loss of connectivity and suitable habitat, which can affect this natural behaviour. In a tributary of River Douro, River Tua, a 108 m high dam, was recently built (i.e., Foz Tua dam in 2017) at only 1.1 km from the river mouth, leaving the upstream spawning area inaccessible. To evaluate the species behavioural response to this impact, a passive acoustic telemetry array was deployed in the study area, and between 2018 and 2023, 120 tagged fish had their movements tracked. The results showed two different behavioural profiles in the same population with migratory (42.5%) and resident (54.0%) individuals. During this period, a specific experimental study was developed to evaluate the response of a subset of 90 fish, captured upstream of the Foz Tua dam and translocated to an alternative downstream Douro tributary (River Pinhão, ~20 km downstream from River Tua). From these, 66.7% remained at the release site, whilst 23.3% returned to the river of origin, i.e., River Tua. From the returned fraction, 28.6% of the tagged fish maintained this migratory behaviour between both rivers in the following years. Generalized Additive Models were used for each of the two behavioural profiles observed in this study, to identify which environmental variables were associated with the presence of the tagged barbels downstream the Foz Tua dam. Ecological flow, temperature and day-period were some of the predictors explaining the use of this river stretch downstream of Foz Tua dam. This study substantially updates the available information regarding this species’ movement patterns at large spatial and temporal scales, contributing to enhancement of management and conservation programs for potamodromous species, in highly impounded and fragmented rivers.

]]> Movement Patterns of the Iberian Barbel (Luciobarbus bocagei Steindachner, 1864) in a Recently Impounded River Ana S. Rato Carlos M. Alexandre Ana F. Silva João P. Marques Sara S. Silva Maria J. Lança Bernardo R. Quintella Pedro R. Almeida doi: 10.3390/proceedings2026146095 Proceedings 2026-06-22 Proceedings 2026-06-22 146 1 Abstract 95 10.3390/proceedings2026146095 https://www.mdpi.com/2504-3900/146/1/95 Proceedings, Vol. 146, Pages 82: The Impact of River Fragmentation on Freshwater Fish Population Connectivity in Spanish River Basins https://www.mdpi.com/2504-3900/146/1/82 Introduction: Artificial infrastructure interrupts river longitudinal connectivity, preventing the free flow of water, matter, energy, and organisms through the system, altering the habitat and impacting freshwater biodiversity. Freshwater fishes are especially sensitive to this threat, since they are constrained to the limits of the river network. Transversal obstacles, such as dams and weirs, hinder their movements upstream and downstream and fragment populations. Longitudinal connectivity can be simply measured as the proportion of connected river length in a basin. However, other indices have been suggested more recently, measuring connectivity as the proportion of connected elements of interest (e.g., populations of a species) in a river basin. Objective: The aim of this work was to study (1) the degree of connectivity of native freshwater fish species populations in eleven Spanish river basins and (2) the impact of artificial river fragmentation in these basins on population connectivity. Methodology: Fish populations’ location and size were estimated through sampling presence data, and completed using the predicted occurrence of each species in a river basin, calculated through Species Distribution Models (SDMs). To estimate the degree of connectivity between populations of each species, the Population Connectivity Index (PCI) was calculated under two scenarios: the “current” scenario, considering all the artificial obstacles fragmenting the river network and their specific passabilities, and the “natural” scenario, considering that all the artificial obstacles in the river network were completely passable. Results: Native freshwater fish populations are severely fragmented in Spanish rivers, with a mean current PCI of 9.8% across species and river basins. The impact of artificial fragmentation is high, causing a mean decrease in PCI of 52 percentage points across species and river basins. Moreover, although the impact of artificial river fragmentation is high in all river basins, it is important to point out that there are significant differences between river basins attributed to their size and the specific traits of the ichthyofauna inhabiting them. Conclusions: The degree of connectivity in a river basin varies depending on the elements of interest considered. Therefore, incorporating ichthyofauna into the decision-making process is essential to improve the effectiveness of river restoration actions. 2026-06-22 Proceedings, Vol. 146, Pages 82: The Impact of River Fragmentation on Freshwater Fish Population Connectivity in Spanish River Basins

Proceedings doi: 10.3390/proceedings2026146082

Authors: Lide Izeta-Zalduendo Rafael Miranda José Barquín Alexia M. González-Ferreras Maria Moran-Luis Francisco J. Peñas Amaia A. Rodeles Ana Sánchez-Alcázar Ana Villarroya David Galicia

Introduction: Artificial infrastructure interrupts river longitudinal connectivity, preventing the free flow of water, matter, energy, and organisms through the system, altering the habitat and impacting freshwater biodiversity. Freshwater fishes are especially sensitive to this threat, since they are constrained to the limits of the river network. Transversal obstacles, such as dams and weirs, hinder their movements upstream and downstream and fragment populations. Longitudinal connectivity can be simply measured as the proportion of connected river length in a basin. However, other indices have been suggested more recently, measuring connectivity as the proportion of connected elements of interest (e.g., populations of a species) in a river basin. Objective: The aim of this work was to study (1) the degree of connectivity of native freshwater fish species populations in eleven Spanish river basins and (2) the impact of artificial river fragmentation in these basins on population connectivity. Methodology: Fish populations’ location and size were estimated through sampling presence data, and completed using the predicted occurrence of each species in a river basin, calculated through Species Distribution Models (SDMs). To estimate the degree of connectivity between populations of each species, the Population Connectivity Index (PCI) was calculated under two scenarios: the “current” scenario, considering all the artificial obstacles fragmenting the river network and their specific passabilities, and the “natural” scenario, considering that all the artificial obstacles in the river network were completely passable. Results: Native freshwater fish populations are severely fragmented in Spanish rivers, with a mean current PCI of 9.8% across species and river basins. The impact of artificial fragmentation is high, causing a mean decrease in PCI of 52 percentage points across species and river basins. Moreover, although the impact of artificial river fragmentation is high in all river basins, it is important to point out that there are significant differences between river basins attributed to their size and the specific traits of the ichthyofauna inhabiting them. Conclusions: The degree of connectivity in a river basin varies depending on the elements of interest considered. Therefore, incorporating ichthyofauna into the decision-making process is essential to improve the effectiveness of river restoration actions.

]]> The Impact of River Fragmentation on Freshwater Fish Population Connectivity in Spanish River Basins Lide Izeta-Zalduendo Rafael Miranda José Barquín Alexia M. González-Ferreras Maria Moran-Luis Francisco J. Peñas Amaia A. Rodeles Ana Sánchez-Alcázar Ana Villarroya David Galicia doi: 10.3390/proceedings2026146082 Proceedings 2026-06-22 Proceedings 2026-06-22 146 1 Abstract 82 10.3390/proceedings2026146082 https://www.mdpi.com/2504-3900/146/1/82 Proceedings, Vol. 146, Pages 93: Invasive Fish Dominance in a Mediterranean Basin: An Updated Inventory from the Segura River (SE Spain) https://www.mdpi.com/2504-3900/146/1/93 Introduction: Freshwater fish communities in Mediterranean basins have undergone profound changes over recent decades due to biological invasions, habitat alteration, and hydrological regulation. The Segura River basin (SE Spain), particularly in the Region of Murcia, represents a paradigmatic case of these transformations, with increasing pressures on native ichthyofauna and growing relevance for recreational fisheries. Objective: This study aims to provide an updated inventory of fish communities in the rivers and reservoirs of the Region of Murcia, assessing current composition, relative abundance, and conservation status. Methodology: Fish assemblages were surveyed during sampling campaigns conducted in autumn 2023, 2024 and 2025. Sampling sites included representative reservoirs and river reaches within the middle sector of the Segura River basin, focusing on areas of interest for recreational fishing. Passive fishing gears were used in reservoirs, while electrofishing was conducted in riverine habitats. Presence and relative abundance data were recorded for all detected taxa. Results: A total of 15 taxa were identified (8 in reservoirs and 15 in river reaches). Reservoir communities were almost entirely dominated by non-native invasive species, including Cyprinus carpio, Alburnus alburnus, Sander lucioperca, Micropterus salmoides, Lepomis gibbosus, and Gambusia holbrooki, with Luciobarbus sclateri as the only native species. Riverine sections of the main channel also showed a strong dominance of exotic taxa, with additional species such as Esox lucius, Oncorhynchus mykiss, Gobio lozanoi, and Pseudochondrostoma polylepis. Native species detected included L. sclateri (dominant), Anguilla anguilla, and Salariopsis fluviatilis, the latter showing a recent expansion likely linked to human-mediated introduction. Isolated tributaries hosted the most valuable assemblages, including populations of L. sclateri, Squalius tartessicus, and the endangered Valencia hispanica (recently unauthorized translocations into the Segura river basin). Conclusions: Current fish communities in the Region of Murcia reflect a marked ecological degradation compared to historical conditions, driven by the proliferation of non-native species and habitat alteration. Changes over the last 30 years have been particularly pronounced in reservoirs and regulated sections of the main river channel, where local extinction of the native S. tartessicus has also been detected. Only the headwaters of certain tributaries retain fish assemblages of notable conservation interest, highlighting their priority for management and protection. 2026-06-22 Proceedings, Vol. 146, Pages 93: Invasive Fish Dominance in a Mediterranean Basin: An Updated Inventory from the Segura River (SE Spain)

Proceedings doi: 10.3390/proceedings2026146093

Authors: Elena Parra-Espín José Manuel Zamora-Marín José Manuel Vidal-Gil Antonio Zamora-López Antonio Guillén-Beltrán Miguel Ángel Richarte Adrián Guerrero-Gómez Antonio Andrés Herrero-Reyes Víctor Manuel Álvarez-Navarro Jorge Madrid-Ruiz Rocío Peñalver Mar Torralva Francisco José Oliva-Paterna

Introduction: Freshwater fish communities in Mediterranean basins have undergone profound changes over recent decades due to biological invasions, habitat alteration, and hydrological regulation. The Segura River basin (SE Spain), particularly in the Region of Murcia, represents a paradigmatic case of these transformations, with increasing pressures on native ichthyofauna and growing relevance for recreational fisheries. Objective: This study aims to provide an updated inventory of fish communities in the rivers and reservoirs of the Region of Murcia, assessing current composition, relative abundance, and conservation status. Methodology: Fish assemblages were surveyed during sampling campaigns conducted in autumn 2023, 2024 and 2025. Sampling sites included representative reservoirs and river reaches within the middle sector of the Segura River basin, focusing on areas of interest for recreational fishing. Passive fishing gears were used in reservoirs, while electrofishing was conducted in riverine habitats. Presence and relative abundance data were recorded for all detected taxa. Results: A total of 15 taxa were identified (8 in reservoirs and 15 in river reaches). Reservoir communities were almost entirely dominated by non-native invasive species, including Cyprinus carpio, Alburnus alburnus, Sander lucioperca, Micropterus salmoides, Lepomis gibbosus, and Gambusia holbrooki, with Luciobarbus sclateri as the only native species. Riverine sections of the main channel also showed a strong dominance of exotic taxa, with additional species such as Esox lucius, Oncorhynchus mykiss, Gobio lozanoi, and Pseudochondrostoma polylepis. Native species detected included L. sclateri (dominant), Anguilla anguilla, and Salariopsis fluviatilis, the latter showing a recent expansion likely linked to human-mediated introduction. Isolated tributaries hosted the most valuable assemblages, including populations of L. sclateri, Squalius tartessicus, and the endangered Valencia hispanica (recently unauthorized translocations into the Segura river basin). Conclusions: Current fish communities in the Region of Murcia reflect a marked ecological degradation compared to historical conditions, driven by the proliferation of non-native species and habitat alteration. Changes over the last 30 years have been particularly pronounced in reservoirs and regulated sections of the main river channel, where local extinction of the native S. tartessicus has also been detected. Only the headwaters of certain tributaries retain fish assemblages of notable conservation interest, highlighting their priority for management and protection.

]]> Invasive Fish Dominance in a Mediterranean Basin: An Updated Inventory from the Segura River (SE Spain) Elena Parra-Espín José Manuel Zamora-Marín José Manuel Vidal-Gil Antonio Zamora-López Antonio Guillén-Beltrán Miguel Ángel Richarte Adrián Guerrero-Gómez Antonio Andrés Herrero-Reyes Víctor Manuel Álvarez-Navarro Jorge Madrid-Ruiz Rocío Peñalver Mar Torralva Francisco José Oliva-Paterna doi: 10.3390/proceedings2026146093 Proceedings 2026-06-22 Proceedings 2026-06-22 146 1 Abstract 93 10.3390/proceedings2026146093 https://www.mdpi.com/2504-3900/146/1/93 Proceedings, Vol. 146, Pages 85: Evaluation of Effluent Water Effects on Cyprinodon variegatus Larvae for the Ecotoxicological Assessment of Endocrine Disruptors Acting on Estrogenic and Androgenic Pathways https://www.mdpi.com/2504-3900/146/1/85 Introduction: The discharge of treated wastewater into coastal and marine environments represents a continuous source of pollutants, including pharmaceuticals and plastic additives with endocrine-disrupting (ED) potential. These compounds are of increasing concern for the European Union due to their capacity to interfere with hormonal systems and their inclusion in current environmental monitoring priorities. ED compounds may induce sublethal effects in aquatic organisms, particularly in vertebrates, where endocrine pathways are highly conserved. In this context, the use of Cyprinodon variegatus, a euryhaline fish species, provides a suitable model to assess potential risks in marine ecosystems. Despite advances in wastewater treatment technologies, the persistence of biologically active substances in treated effluents remains a concern. Objective: This study aims to evaluate whether treated effluent water still contains compounds with endocrine-disrupting activity and to assess their potential effects on marine organisms. Methodology: Larvae of C. variegatus from a laboratory stock maintained at ECIMAT (University of Vigo), one of the few available stocks of this species in Europe, were exposed for 48 h to environmentally relevant dilutions (1:10, 1:30, and 1:100) of wastewater treatment plant effluent collected after UV disinfection as the final treatment step. Pools of 10 larvae were used for each condition. Sublethal effects were assessed through gene expression analysis using quantitative PCR (qPCR), targeting biomarkers involved in endocrine regulation. Two housekeeping genes (tbp and hprt) were used for normalization. Estrogenic responses were evaluated through vtgab and zp2, while androgenic responses were assessed using 17hsd and 11hsd. Results: Preliminary results indicate significant alterations in estrogen-related gene expression, particularly in vitellogenin (vtgab) and zona pellucida (zp2), highlighting the activation of estrogenic pathways and supporting the presence of endocrine-disrupting activity in treated effluent water. Conclusions: This study highlights the relevance of assessing endocrine disrupting activity in treated effluents and supports the use of molecular biomarkers as sensitive tools for evaluating their potential impact on marine ecosystems, contributing to the improvement of wastewater monitoring and management strategies. 2026-06-22 Proceedings, Vol. 146, Pages 85: Evaluation of Effluent Water Effects on Cyprinodon variegatus Larvae for the Ecotoxicological Assessment of Endocrine Disruptors Acting on Estrogenic and Androgenic Pathways

Proceedings doi: 10.3390/proceedings2026146085

Authors: Raquel Abad Antía Alonso Alexandre M. Schönemann Humberto Quesada Ricardo Beiras

Introduction: The discharge of treated wastewater into coastal and marine environments represents a continuous source of pollutants, including pharmaceuticals and plastic additives with endocrine-disrupting (ED) potential. These compounds are of increasing concern for the European Union due to their capacity to interfere with hormonal systems and their inclusion in current environmental monitoring priorities. ED compounds may induce sublethal effects in aquatic organisms, particularly in vertebrates, where endocrine pathways are highly conserved. In this context, the use of Cyprinodon variegatus, a euryhaline fish species, provides a suitable model to assess potential risks in marine ecosystems. Despite advances in wastewater treatment technologies, the persistence of biologically active substances in treated effluents remains a concern. Objective: This study aims to evaluate whether treated effluent water still contains compounds with endocrine-disrupting activity and to assess their potential effects on marine organisms. Methodology: Larvae of C. variegatus from a laboratory stock maintained at ECIMAT (University of Vigo), one of the few available stocks of this species in Europe, were exposed for 48 h to environmentally relevant dilutions (1:10, 1:30, and 1:100) of wastewater treatment plant effluent collected after UV disinfection as the final treatment step. Pools of 10 larvae were used for each condition. Sublethal effects were assessed through gene expression analysis using quantitative PCR (qPCR), targeting biomarkers involved in endocrine regulation. Two housekeeping genes (tbp and hprt) were used for normalization. Estrogenic responses were evaluated through vtgab and zp2, while androgenic responses were assessed using 17hsd and 11hsd. Results: Preliminary results indicate significant alterations in estrogen-related gene expression, particularly in vitellogenin (vtgab) and zona pellucida (zp2), highlighting the activation of estrogenic pathways and supporting the presence of endocrine-disrupting activity in treated effluent water. Conclusions: This study highlights the relevance of assessing endocrine disrupting activity in treated effluents and supports the use of molecular biomarkers as sensitive tools for evaluating their potential impact on marine ecosystems, contributing to the improvement of wastewater monitoring and management strategies.

]]> Evaluation of Effluent Water Effects on Cyprinodon variegatus Larvae for the Ecotoxicological Assessment of Endocrine Disruptors Acting on Estrogenic and Androgenic Pathways Raquel Abad Antía Alonso Alexandre M. Schönemann Humberto Quesada Ricardo Beiras doi: 10.3390/proceedings2026146085 Proceedings 2026-06-22 Proceedings 2026-06-22 146 1 Abstract 85 10.3390/proceedings2026146085 https://www.mdpi.com/2504-3900/146/1/85 Proceedings, Vol. 146, Pages 83: Baseline Elemental Profile of Juvenile Sharks from a Multispecies Nursery Area off West Africa (Sal Rei Bay, Boa Vista Island, Cabo Verde) https://www.mdpi.com/2504-3900/146/1/83 Introduction: Establishing baseline descriptions of inorganic elements in the early life stages of sharks and in their respective nursery areas is essential for assessing anthropogenic impacts and supporting conservation strategies. Objectives: This study presents the first baseline of plasma trace element concentrations (Al, Zn, As, Cu, Cr, Cd, Co, Mn, Ti, Ni, Hg, Pb) for four juvenile shark species (Carcharhinus limbatus, Paragaleus pectoralis, Rhizoprionodon acutus, and Sphyrna lewini) from Sal Rei Bay, Boa Vista Island, Cabo Verde—the first multi-species shark nursery area described in Atlantic Africa. Methodology: Seawater and sediment samples were collected from eight sites and analyzed along with plasma samples using total reflection X-ray fluorescence spectroscopy (TXRF). Sediment granulometry and pollution indices, including the enrichment factor (EF), ecological risk index (RI), and metal pollution index (MPI), were used to characterize habitat contamination. Data were analyzed using statistical models to explore spatial and element-specific patterns. Results: Overall, environmental contamination was low, with slight increases in Cd, Co, and Hg at sites 1 and 2, near the fishing port, and at site 5, likely reflecting natural transport, sediment redistribution, and enhanced nearshore deposition. Juvenile sharks exhibited generally low plasma trace element concentrations, although species-specific elemental signatures were evident: elevated levels of Al and Cu in C. limbatus, Zn in S. lewini, and As in R. acutus and P. pectoralis. Conclusions: These findings establish critical baseline reference values for trace elements in juvenile sharks from a key Atlantic nursery area. The results provide an essential framework for future biomonitoring efforts and contribute to the management and conservation of Cabo Verdean shark nursery habitats. 2026-06-22 Proceedings, Vol. 146, Pages 83: Baseline Elemental Profile of Juvenile Sharks from a Multispecies Nursery Area off West Africa (Sal Rei Bay, Boa Vista Island, Cabo Verde)

Proceedings doi: 10.3390/proceedings2026146083

Authors: Marta Ramalho Catarina Caldeira-Santos Melanie Court Jaquelino Varela Bernardo Duarte Rui Rosa

Introduction: Establishing baseline descriptions of inorganic elements in the early life stages of sharks and in their respective nursery areas is essential for assessing anthropogenic impacts and supporting conservation strategies. Objectives: This study presents the first baseline of plasma trace element concentrations (Al, Zn, As, Cu, Cr, Cd, Co, Mn, Ti, Ni, Hg, Pb) for four juvenile shark species (Carcharhinus limbatus, Paragaleus pectoralis, Rhizoprionodon acutus, and Sphyrna lewini) from Sal Rei Bay, Boa Vista Island, Cabo Verde—the first multi-species shark nursery area described in Atlantic Africa. Methodology: Seawater and sediment samples were collected from eight sites and analyzed along with plasma samples using total reflection X-ray fluorescence spectroscopy (TXRF). Sediment granulometry and pollution indices, including the enrichment factor (EF), ecological risk index (RI), and metal pollution index (MPI), were used to characterize habitat contamination. Data were analyzed using statistical models to explore spatial and element-specific patterns. Results: Overall, environmental contamination was low, with slight increases in Cd, Co, and Hg at sites 1 and 2, near the fishing port, and at site 5, likely reflecting natural transport, sediment redistribution, and enhanced nearshore deposition. Juvenile sharks exhibited generally low plasma trace element concentrations, although species-specific elemental signatures were evident: elevated levels of Al and Cu in C. limbatus, Zn in S. lewini, and As in R. acutus and P. pectoralis. Conclusions: These findings establish critical baseline reference values for trace elements in juvenile sharks from a key Atlantic nursery area. The results provide an essential framework for future biomonitoring efforts and contribute to the management and conservation of Cabo Verdean shark nursery habitats.

]]> Baseline Elemental Profile of Juvenile Sharks from a Multispecies Nursery Area off West Africa (Sal Rei Bay, Boa Vista Island, Cabo Verde) Marta Ramalho Catarina Caldeira-Santos Melanie Court Jaquelino Varela Bernardo Duarte Rui Rosa doi: 10.3390/proceedings2026146083 Proceedings 2026-06-22 Proceedings 2026-06-22 146 1 Abstract 83 10.3390/proceedings2026146083 https://www.mdpi.com/2504-3900/146/1/83 Proceedings, Vol. 146, Pages 94: How Many Native Brown Trout Species Are There Within the Iberian Peninsula? https://www.mdpi.com/2504-3900/146/1/94 Introduction: Brown trout (Salmo trutta L.) is a salmonid fish considered a genetically and geographically highly structured species. The species is catalogued as vulnerable in the Spanish Red List since 1992. Based on morphological differentiation, some authors have proposed splitting Salmo trutta into many taxa and in recent years, several binomials involving morphologically differentiated Western Mediterranean brown trout morphs have regained relevance as distinct species and are now assessed in the IUCN Red List. However, Spanish legislation still considers a single species, S. trutta, inhabiting Atlantic and Mediterranean rivers. Analyses of mitochondrial DNA variation across the species’ native range have revealed numerous matrilineal lineages and helped disentangle the phylogenetic relationships and biogeographic history of brown trout. Four largely distant evolutionary lineages defined by sequencing the mitochondrial control region (Adriatic, AD, Mediterranean, ME, Atlantic, AT and Duero, DU) are native among the Iberian brown trout populations. Stocking activities have resulted in introgressive hybridization of these native populations with non-native northern Atlantic stocks throughout the Iberian rivers and compromise the evolutionary singularities of brown trout in the Iberian Peninsula. Methodology: In this study, we compare complete mitogenomes from Iberian brown trout specimens with those from other regions, using both newly sequenced samples and whole mitogenomes from the GenBank database and estimate their divergence times relative to those observed among species of other Salmoninae genera, such as Oncorhynchus and Salvelinus, to assess patterns of species-specific divergence among Iberian brown trout populations. Results: The evolutionary relationships of the specimens using their mitogenomes fitted with previous analyses using partial mitochondrial sequences and confirmed the strong differentiation among the Iberian lineages. All the branches involving distant Iberian mitogenomes are sister branches with taxa historically described under different binomials (such as Salmo macrostigma, S. multipunctatus, S. pallaryi). Conclusions: The results provide evidence supporting the existence of potentially distinct and undescribed native Salmo species in the Iberian rivers. 2026-06-22 Proceedings, Vol. 146, Pages 94: How Many Native Brown Trout Species Are There Within the Iberian Peninsula?

Proceedings doi: 10.3390/proceedings2026146094

Authors: Álvaro Pita Manuel Vera Sandra Heras Jordi Morata Raúl Tonda David -Diez-del-Molino Dorte Bekkevold Adrián Casanova Alba Abras Maria Inés Roldán Jose-Luis García-Marin

Introduction: Brown trout (Salmo trutta L.) is a salmonid fish considered a genetically and geographically highly structured species. The species is catalogued as vulnerable in the Spanish Red List since 1992. Based on morphological differentiation, some authors have proposed splitting Salmo trutta into many taxa and in recent years, several binomials involving morphologically differentiated Western Mediterranean brown trout morphs have regained relevance as distinct species and are now assessed in the IUCN Red List. However, Spanish legislation still considers a single species, S. trutta, inhabiting Atlantic and Mediterranean rivers. Analyses of mitochondrial DNA variation across the species’ native range have revealed numerous matrilineal lineages and helped disentangle the phylogenetic relationships and biogeographic history of brown trout. Four largely distant evolutionary lineages defined by sequencing the mitochondrial control region (Adriatic, AD, Mediterranean, ME, Atlantic, AT and Duero, DU) are native among the Iberian brown trout populations. Stocking activities have resulted in introgressive hybridization of these native populations with non-native northern Atlantic stocks throughout the Iberian rivers and compromise the evolutionary singularities of brown trout in the Iberian Peninsula. Methodology: In this study, we compare complete mitogenomes from Iberian brown trout specimens with those from other regions, using both newly sequenced samples and whole mitogenomes from the GenBank database and estimate their divergence times relative to those observed among species of other Salmoninae genera, such as Oncorhynchus and Salvelinus, to assess patterns of species-specific divergence among Iberian brown trout populations. Results: The evolutionary relationships of the specimens using their mitogenomes fitted with previous analyses using partial mitochondrial sequences and confirmed the strong differentiation among the Iberian lineages. All the branches involving distant Iberian mitogenomes are sister branches with taxa historically described under different binomials (such as Salmo macrostigma, S. multipunctatus, S. pallaryi). Conclusions: The results provide evidence supporting the existence of potentially distinct and undescribed native Salmo species in the Iberian rivers.

]]> How Many Native Brown Trout Species Are There Within the Iberian Peninsula? Álvaro Pita Manuel Vera Sandra Heras Jordi Morata Raúl Tonda David -Diez-del-Molino Dorte Bekkevold Adrián Casanova Alba Abras Maria Inés Roldán Jose-Luis García-Marin doi: 10.3390/proceedings2026146094 Proceedings 2026-06-22 Proceedings 2026-06-22 146 1 Abstract 94 10.3390/proceedings2026146094 https://www.mdpi.com/2504-3900/146/1/94 Proceedings, Vol. 146, Pages 92: Seven Years of Citizen Science Reveal Spatial and Seasonal Priorities for Shark and Batoid Conservation in the Central Maldives https://www.mdpi.com/2504-3900/146/1/92 Introduction: Elasmobranchs play a vital role in marine food webs through top-down control and the structuring of ecosystem stability, yet more than one-third of species face extinction. The Maldives, a recognised Indian Ocean hotspot for shark and batoid diversity, designated its EEZ as a shark sanctuary in 2010, but multispecies elasmobranch occurrence patterns and environmental drivers remain poorly characterised in Lhaviyani Atoll in the central Maldives, which hosts two Important Shark and Ray Areas (ISRAs). Recreational SCUBA networks can turn routine dive activity into long-term conservation evidence, already informing nearly 10% of the western Indian Ocean ISRAs. Objective: To characterise spatiotemporal patterns of elasmobranch assemblages in Lhaviyani Atoll (2017–2024), quantify how environmental and geomorphic drivers shape relative abundance, diversity, and hotspots, and provide evidence for targeted elasmobranch conservation. Methodology: A seven-year opportunistic dive-log dataset of 12,732 SCUBA surveys and 142,994 elasmobranch records across 94 dive sites was analysed. Effort-standardised relative abundance and community metrics (Shannon diversity, Pielou’s evenness) were modelled against sea surface temperature (SST), salinity, dissolved oxygen, chlorophyll-a, zonal current velocity, substrate type, and reef geomorphology using generalised additive models (GAMs). Spatial analyses identified persistent northern-rim aggregation areas aligned with ISRAs. Results: Twenty-eight species (14 sharks, 14 batoids) were recorded, including 23 threatened on the IUCN Red List (4 Critically Endangered, 12 Endangered, 7 Vulnerable). Relative abundance and diversity peaked during the late southwest monsoon (August–September) and declined during the northeast monsoon (December–March). After 2021, diversity and evenness increased while overall abundance declined. Relative abundance was primarily driven by SST, salinity, and current velocity; for sharks, dissolved oxygen and chlorophyll-a were additionally significant, whereas batoid abundance was driven mainly by temperature, oxygen, and current velocity. Four persistent hotspots along the northern atoll rim were identified, with sharks concentrated along exposed slopes and channels, and batoids distributed broadly within lagoonal habitats. Conclusions: Long-term citizen science dive-log monitoring is cost-effective for elasmobranch conservation in remote tropical seascapes. These results show how dive-industry partnerships can inform conservation governance over a decade after sanctuary designation, supporting targeted, habitat-focused management as shark and batoid conservation frameworks continue to evolve. 2026-06-22 Proceedings, Vol. 146, Pages 92: Seven Years of Citizen Science Reveal Spatial and Seasonal Priorities for Shark and Batoid Conservation in the Central Maldives

Proceedings doi: 10.3390/proceedings2026146092

Authors: Margarida Vizeu-Pinheiro Sebastião Farias Maria Lourie Saoirse Tak-Yung Macklin Paula Dominguez Rein-Loring Ray van Eeden Rui Rosa

Introduction: Elasmobranchs play a vital role in marine food webs through top-down control and the structuring of ecosystem stability, yet more than one-third of species face extinction. The Maldives, a recognised Indian Ocean hotspot for shark and batoid diversity, designated its EEZ as a shark sanctuary in 2010, but multispecies elasmobranch occurrence patterns and environmental drivers remain poorly characterised in Lhaviyani Atoll in the central Maldives, which hosts two Important Shark and Ray Areas (ISRAs). Recreational SCUBA networks can turn routine dive activity into long-term conservation evidence, already informing nearly 10% of the western Indian Ocean ISRAs. Objective: To characterise spatiotemporal patterns of elasmobranch assemblages in Lhaviyani Atoll (2017–2024), quantify how environmental and geomorphic drivers shape relative abundance, diversity, and hotspots, and provide evidence for targeted elasmobranch conservation. Methodology: A seven-year opportunistic dive-log dataset of 12,732 SCUBA surveys and 142,994 elasmobranch records across 94 dive sites was analysed. Effort-standardised relative abundance and community metrics (Shannon diversity, Pielou’s evenness) were modelled against sea surface temperature (SST), salinity, dissolved oxygen, chlorophyll-a, zonal current velocity, substrate type, and reef geomorphology using generalised additive models (GAMs). Spatial analyses identified persistent northern-rim aggregation areas aligned with ISRAs. Results: Twenty-eight species (14 sharks, 14 batoids) were recorded, including 23 threatened on the IUCN Red List (4 Critically Endangered, 12 Endangered, 7 Vulnerable). Relative abundance and diversity peaked during the late southwest monsoon (August–September) and declined during the northeast monsoon (December–March). After 2021, diversity and evenness increased while overall abundance declined. Relative abundance was primarily driven by SST, salinity, and current velocity; for sharks, dissolved oxygen and chlorophyll-a were additionally significant, whereas batoid abundance was driven mainly by temperature, oxygen, and current velocity. Four persistent hotspots along the northern atoll rim were identified, with sharks concentrated along exposed slopes and channels, and batoids distributed broadly within lagoonal habitats. Conclusions: Long-term citizen science dive-log monitoring is cost-effective for elasmobranch conservation in remote tropical seascapes. These results show how dive-industry partnerships can inform conservation governance over a decade after sanctuary designation, supporting targeted, habitat-focused management as shark and batoid conservation frameworks continue to evolve.

]]> Seven Years of Citizen Science Reveal Spatial and Seasonal Priorities for Shark and Batoid Conservation in the Central Maldives Margarida Vizeu-Pinheiro Sebastião Farias Maria Lourie Saoirse Tak-Yung Macklin Paula Dominguez Rein-Loring Ray van Eeden Rui Rosa doi: 10.3390/proceedings2026146092 Proceedings 2026-06-22 Proceedings 2026-06-22 146 1 Abstract 92 10.3390/proceedings2026146092 https://www.mdpi.com/2504-3900/146/1/92 Proceedings, Vol. 146, Pages 88: Advancing the Quality Diagnosis and Monitoring of Aquatic Pollution https://www.mdpi.com/2504-3900/146/1/88 Introduction: Aquatic chemical pollution is among the most worrying threats to ecosystem health. There is an ever-increasing variety of pollutant substances detected across the source-to-sea continuum, causing loss of biodiversity and ecological disequilibrium. Achieving cleaner and healthier systems relies on carrying out sustained, cost-effective, diagnosis and aquatic effects monitoring, within the adaptive management cycle. The available methods are, however, cumbersome, which creates a clear need for innovative expeditious approaches for low-cost surveillance monitoring. In the last decade, Raman Spectroscopy (RS) has gained wide recognition for application to biological questions, for its ability to uncover the complexity of molecules and their interactions. Various fields, from pharmacology to disease diagnosis and prognosis, have suffered an innovation revolution through the application of RS. In this technique inelastic light scattering of a small part of photons of an incident electromagnetic monochromatic light beam (ranging from near-infrared to visible or ultraviolet) is caused by the molecular vibration of chemical bonds. This results in shifts in energy, which indicate discrete vibrational modes of polarisable molecules, providing qualitative and quantitative assessments of the chemical composition and molecular structure of the sample. The technique shows high sensitivity, no need for sample preparation and the possibility of use in non-invasive and label-free analysis. Objective: The aim of this work is to present and discuss evidence about the application of Raman Spectroscopy (RS) to environmental diagnosis and aquatic effect monitoring of pollution. Methodology: The technique was applied to different biological models, i.e., diatoms, zebrafish embryos and larvae and freshwater snails. Quality assessments with diatoms were tested in environmental monitoring, while assessments with other models were done upon exposure to metals and organic contaminants. Results and conclusions: The Raman spectra obtained from the samples analysed comprised bands detected within the 800 to 2000 cm−1 wavenumber range. These were related to bond vibrations of carbohydrates, DNA phosphate groups, proteins or CH, NH and OH stretching in lipids and proteins. Data analysis using chemometric methods clearly distinguished pollutant exposure from control sites or treatments, pointing out the potential for surveyance monitoring. The next steps include the comparison with other sensitive methods (e.g., locomotion and avoidance behaviours, omics methods) to assess efficiency and bring further mechanistic understanding. 2026-06-22 Proceedings, Vol. 146, Pages 88: Advancing the Quality Diagnosis and Monitoring of Aquatic Pollution

Proceedings doi: 10.3390/proceedings2026146088

Authors: Laura Guimarães Luís Oliva-Teles Raquel Pinto Cláudia Teixeira Pedro Rodrigues Matilde Moreira-Santos António Paulo Carvalho

Introduction: Aquatic chemical pollution is among the most worrying threats to ecosystem health. There is an ever-increasing variety of pollutant substances detected across the source-to-sea continuum, causing loss of biodiversity and ecological disequilibrium. Achieving cleaner and healthier systems relies on carrying out sustained, cost-effective, diagnosis and aquatic effects monitoring, within the adaptive management cycle. The available methods are, however, cumbersome, which creates a clear need for innovative expeditious approaches for low-cost surveillance monitoring. In the last decade, Raman Spectroscopy (RS) has gained wide recognition for application to biological questions, for its ability to uncover the complexity of molecules and their interactions. Various fields, from pharmacology to disease diagnosis and prognosis, have suffered an innovation revolution through the application of RS. In this technique inelastic light scattering of a small part of photons of an incident electromagnetic monochromatic light beam (ranging from near-infrared to visible or ultraviolet) is caused by the molecular vibration of chemical bonds. This results in shifts in energy, which indicate discrete vibrational modes of polarisable molecules, providing qualitative and quantitative assessments of the chemical composition and molecular structure of the sample. The technique shows high sensitivity, no need for sample preparation and the possibility of use in non-invasive and label-free analysis. Objective: The aim of this work is to present and discuss evidence about the application of Raman Spectroscopy (RS) to environmental diagnosis and aquatic effect monitoring of pollution. Methodology: The technique was applied to different biological models, i.e., diatoms, zebrafish embryos and larvae and freshwater snails. Quality assessments with diatoms were tested in environmental monitoring, while assessments with other models were done upon exposure to metals and organic contaminants. Results and conclusions: The Raman spectra obtained from the samples analysed comprised bands detected within the 800 to 2000 cm−1 wavenumber range. These were related to bond vibrations of carbohydrates, DNA phosphate groups, proteins or CH, NH and OH stretching in lipids and proteins. Data analysis using chemometric methods clearly distinguished pollutant exposure from control sites or treatments, pointing out the potential for surveyance monitoring. The next steps include the comparison with other sensitive methods (e.g., locomotion and avoidance behaviours, omics methods) to assess efficiency and bring further mechanistic understanding.

]]> Advancing the Quality Diagnosis and Monitoring of Aquatic Pollution Laura Guimarães Luís Oliva-Teles Raquel Pinto Cláudia Teixeira Pedro Rodrigues Matilde Moreira-Santos António Paulo Carvalho doi: 10.3390/proceedings2026146088 Proceedings 2026-06-22 Proceedings 2026-06-22 146 1 Abstract 88 10.3390/proceedings2026146088 https://www.mdpi.com/2504-3900/146/1/88 Proceedings, Vol. 146, Pages 89: Comparative Osteology of Species of Genus Thorichthys Meek 1904 (Cichliformes: Cichlidae) https://www.mdpi.com/2504-3900/146/1/89 Introduction: The genus Thorichthys comprises nine species of Central American freshwater cichlids, which are distributed along the eastern slopes of Mexico and Honduras. These species are grouped into two subgroups: “helleri” and “meeki”. While some analyses of the group’s ecomorphology and character evolution have been conducted, the skeletal structures of this genus remain to be described. Objective: This study provides an osteological description of the species in the genus Thorichthys. Methodology: The specimens were sectioned and stained with alizarin red, and the bone structures were analyzed and described. Results: The genus Thorichthys possesses 12 abdominal vertebrae and 12–13 caudal vertebrae. Species of “helleri” group are characterized by the following points: (1) there is a urohyal bone with a sharply concave dorsal margin and a spiniform, inclined process; (2) typically, there are a total of 14–15 gill rakers in the first gill arch; (3) the opercular bone bears a dorsally displaced, posterior protuberance; (4) a total of 12–13 anal pterygiophores; and (5) a slightly recurved cephalic profile. Meanwhile species of the “meeki” group exhibit the following characteristics: (1) a urohyal with a broad, unsharpened dorsal concavity and a small, blunt, nearly straight process; (2) a total of 15–21 gill spines; (3) a dorsally positioned protuberance in the middle of the opercular bone; (4) a total of 14–16 proximal anal pterygiophores; and (5) a straight cephalic profile. Conclusions: These bony elements are proposed as a complement to the taxonomic differentiation of the species in Thorichthys, and to contribute new morphological characteristics to the study of other cichlid genera. 2026-06-22 Proceedings, Vol. 146, Pages 89: Comparative Osteology of Species of Genus Thorichthys Meek 1904 (Cichliformes: Cichlidae)

Proceedings doi: 10.3390/proceedings2026146089

Authors: Eduardo López-Segovia Juan J. Schmitter-Soto Jesús Alvarado-Ortega Luis Fernando Del Moral-Flores

Introduction: The genus Thorichthys comprises nine species of Central American freshwater cichlids, which are distributed along the eastern slopes of Mexico and Honduras. These species are grouped into two subgroups: “helleri” and “meeki”. While some analyses of the group’s ecomorphology and character evolution have been conducted, the skeletal structures of this genus remain to be described. Objective: This study provides an osteological description of the species in the genus Thorichthys. Methodology: The specimens were sectioned and stained with alizarin red, and the bone structures were analyzed and described. Results: The genus Thorichthys possesses 12 abdominal vertebrae and 12–13 caudal vertebrae. Species of “helleri” group are characterized by the following points: (1) there is a urohyal bone with a sharply concave dorsal margin and a spiniform, inclined process; (2) typically, there are a total of 14–15 gill rakers in the first gill arch; (3) the opercular bone bears a dorsally displaced, posterior protuberance; (4) a total of 12–13 anal pterygiophores; and (5) a slightly recurved cephalic profile. Meanwhile species of the “meeki” group exhibit the following characteristics: (1) a urohyal with a broad, unsharpened dorsal concavity and a small, blunt, nearly straight process; (2) a total of 15–21 gill spines; (3) a dorsally positioned protuberance in the middle of the opercular bone; (4) a total of 14–16 proximal anal pterygiophores; and (5) a straight cephalic profile. Conclusions: These bony elements are proposed as a complement to the taxonomic differentiation of the species in Thorichthys, and to contribute new morphological characteristics to the study of other cichlid genera.

]]> Comparative Osteology of Species of Genus Thorichthys Meek 1904 (Cichliformes: Cichlidae) Eduardo López-Segovia Juan J. Schmitter-Soto Jesús Alvarado-Ortega Luis Fernando Del Moral-Flores doi: 10.3390/proceedings2026146089 Proceedings 2026-06-22 Proceedings 2026-06-22 146 1 Abstract 89 10.3390/proceedings2026146089 https://www.mdpi.com/2504-3900/146/1/89 Proceedings, Vol. 146, Pages 90: Prioritizing Sites for Fish Translocation Actions: Developing a Fragmentation Index for the Conservation of Diadromous Species https://www.mdpi.com/2504-3900/146/1/90 Introduction: Restoring riverine connectivity is a cornerstone of ecological restoration for migratory fish populations. When physical barriers like dams lack effective fishways, translocation to more suitable sites becomes an alternative. Objectives: This study aims to present a decision-support methodology based on the Fragmentation Index (FI), designed to prioritize release sites in alternative river stretches that maximize the likelihood of survival of translocated diadromous fish. Methodology: The method integrates field-based obstacle characterization and transposability classification, together with a weighted penalty for restrictive obstacles located closer to the confluence with the main stem. The methodology was applied to six tributaries of the Douro River, targeting the European eel (Anguilla anguilla). Results: The FI successfully distinguished between functional reaches and severely fragmented systems. Results revealed high heterogeneity among the studied tributaries, with the Távora (FI = 1.07) and Ceira (FI = 1.12) Rivers identified as top priorities due to low fragmentation and stable hydrology. In contrast, the Tedo River (FI = 5.18) illustrates index’s sensitivity. Despite a high barrier density, its downstream stretch of ~14 km remains functionally connected because the first restrictive obstacles are located far upstream from the confluence. Conversely, the Torto River (FI = 0) was excluded due to severe drought conditions, underscoring the need to pair connectivity metrics with hydrologic viability. Conclusions: For large-scale translocations, this framework enables distributing fish across multiple systems to safeguard the ecological integrity of recipient communities while ensuring individuals can successfully complete their life cycles. Overall, this approach provides a quantitative and replicable framework for managing endangered species by prioritizing release sites with high longitudinal connectivity. 2026-06-22 Proceedings, Vol. 146, Pages 90: Prioritizing Sites for Fish Translocation Actions: Developing a Fragmentation Index for the Conservation of Diadromous Species

Proceedings doi: 10.3390/proceedings2026146090

Authors: Marta Ramalho Ana S. Rato Carlos M. Alexandre Bernardo R. Quintella Pedro R. Almeida

Introduction: Restoring riverine connectivity is a cornerstone of ecological restoration for migratory fish populations. When physical barriers like dams lack effective fishways, translocation to more suitable sites becomes an alternative. Objectives: This study aims to present a decision-support methodology based on the Fragmentation Index (FI), designed to prioritize release sites in alternative river stretches that maximize the likelihood of survival of translocated diadromous fish. Methodology: The method integrates field-based obstacle characterization and transposability classification, together with a weighted penalty for restrictive obstacles located closer to the confluence with the main stem. The methodology was applied to six tributaries of the Douro River, targeting the European eel (Anguilla anguilla). Results: The FI successfully distinguished between functional reaches and severely fragmented systems. Results revealed high heterogeneity among the studied tributaries, with the Távora (FI = 1.07) and Ceira (FI = 1.12) Rivers identified as top priorities due to low fragmentation and stable hydrology. In contrast, the Tedo River (FI = 5.18) illustrates index’s sensitivity. Despite a high barrier density, its downstream stretch of ~14 km remains functionally connected because the first restrictive obstacles are located far upstream from the confluence. Conversely, the Torto River (FI = 0) was excluded due to severe drought conditions, underscoring the need to pair connectivity metrics with hydrologic viability. Conclusions: For large-scale translocations, this framework enables distributing fish across multiple systems to safeguard the ecological integrity of recipient communities while ensuring individuals can successfully complete their life cycles. Overall, this approach provides a quantitative and replicable framework for managing endangered species by prioritizing release sites with high longitudinal connectivity.

]]> Prioritizing Sites for Fish Translocation Actions: Developing a Fragmentation Index for the Conservation of Diadromous Species Marta Ramalho Ana S. Rato Carlos M. Alexandre Bernardo R. Quintella Pedro R. Almeida doi: 10.3390/proceedings2026146090 Proceedings 2026-06-22 Proceedings 2026-06-22 146 1 Abstract 90 10.3390/proceedings2026146090 https://www.mdpi.com/2504-3900/146/1/90 Proceedings, Vol. 146, Pages 84: Monitoring and Mitigation of Migratory Fish Accumulation Influx Downstream of the Foz Tua Dam https://www.mdpi.com/2504-3900/146/1/84 The Tua River is a tributary of the Douro River in the North of Portugal used as a spawning ground for potamodromous fish, namely the Iberian barbel (Luciobarbus bocagei, Steindachner, 1864). Although access to this tributary became severely constrained after the construction of the Foz Tua Hydroelectric Facility (AHFT), fish continued to use the remaining accessible 1.1 km stretch of the Tua River below the dam, especially during their spawning season. Therefore, this study presents the monitoring of migratory fish influx downstream of the AHFT and associated mitigation measures. Fixed and mobile surveys, using an ARIS 1800 sonar, and focused on Iberian barbel were conducted between March and July, from 2023 to 2025. In 2023, fixed sonar monitoring recorded 100,289 individuals, showing a progressive increase over the sampling period, while mobile surveys confirmed high local concentrations (2083 individuals) and temporal fluctuations. In 2024, total counts rose substantially to 182,216 individuals (fixed surveys) and 2656 individuals (mobile surveys), with a peak in early May followed by a gradual reduction in these numbers. In 2025, the highest abundance was observed, with 196,935 individuals (fixed surveys) and 5441 individuals (mobile surveys), alongside higher variability between monitoring campaigns. Overall, these results suggest an intensifying pattern of fish accumulation downstream of the AHFT during the sampled periods, with recurring seasonal peaks. As a method to mitigate massive accumulation of fish downstream of this dam, in 2024 and 2025, a near real-time detection and mitigation protocol was implemented. This protocol identifies an initial “trigger” and a sequential methodology that recognizes possible massive accumulation scenarios, followed by the application of an adaptive operational management measure (e.g., ecological flow regulation) by the AHFT. The application of these measures effectively contributed to reducing fish accumulation during the critical periods. In conclusion, the results highlight a consistent increase in migratory fish accumulation, over the study period, downstream of the AHFT. The successful application of adaptive measures demonstrates that the implemented strategy seems to be effective so far and provides a strong basis for future management actions. 2026-06-22 Proceedings, Vol. 146, Pages 84: Monitoring and Mitigation of Migratory Fish Accumulation Influx Downstream of the Foz Tua Dam

Proceedings doi: 10.3390/proceedings2026146084

Authors: Ana Beatriz Oliveira Ana Sofia Rato Carlos M. Alexandre Rita Almeida Maria João Lança Bernardo R. Quintella Pedro R. Almeida

The Tua River is a tributary of the Douro River in the North of Portugal used as a spawning ground for potamodromous fish, namely the Iberian barbel (Luciobarbus bocagei, Steindachner, 1864). Although access to this tributary became severely constrained after the construction of the Foz Tua Hydroelectric Facility (AHFT), fish continued to use the remaining accessible 1.1 km stretch of the Tua River below the dam, especially during their spawning season. Therefore, this study presents the monitoring of migratory fish influx downstream of the AHFT and associated mitigation measures. Fixed and mobile surveys, using an ARIS 1800 sonar, and focused on Iberian barbel were conducted between March and July, from 2023 to 2025. In 2023, fixed sonar monitoring recorded 100,289 individuals, showing a progressive increase over the sampling period, while mobile surveys confirmed high local concentrations (2083 individuals) and temporal fluctuations. In 2024, total counts rose substantially to 182,216 individuals (fixed surveys) and 2656 individuals (mobile surveys), with a peak in early May followed by a gradual reduction in these numbers. In 2025, the highest abundance was observed, with 196,935 individuals (fixed surveys) and 5441 individuals (mobile surveys), alongside higher variability between monitoring campaigns. Overall, these results suggest an intensifying pattern of fish accumulation downstream of the AHFT during the sampled periods, with recurring seasonal peaks. As a method to mitigate massive accumulation of fish downstream of this dam, in 2024 and 2025, a near real-time detection and mitigation protocol was implemented. This protocol identifies an initial “trigger” and a sequential methodology that recognizes possible massive accumulation scenarios, followed by the application of an adaptive operational management measure (e.g., ecological flow regulation) by the AHFT. The application of these measures effectively contributed to reducing fish accumulation during the critical periods. In conclusion, the results highlight a consistent increase in migratory fish accumulation, over the study period, downstream of the AHFT. The successful application of adaptive measures demonstrates that the implemented strategy seems to be effective so far and provides a strong basis for future management actions.

]]> Monitoring and Mitigation of Migratory Fish Accumulation Influx Downstream of the Foz Tua Dam Ana Beatriz Oliveira Ana Sofia Rato Carlos M. Alexandre Rita Almeida Maria João Lança Bernardo R. Quintella Pedro R. Almeida doi: 10.3390/proceedings2026146084 Proceedings 2026-06-22 Proceedings 2026-06-22 146 1 Abstract 84 10.3390/proceedings2026146084 https://www.mdpi.com/2504-3900/146/1/84 Proceedings, Vol. 146, Pages 87: Identifying Priority Conservation Areas for Iberian Freshwater Fish: National vs. Transboundary Approach https://www.mdpi.com/2504-3900/146/1/87 Introduction: Freshwater habitats underpin global biodiversity and provide an array of essential ecosystem services to humans. However, threat hotspots like the Iberian Peninsula combine severe anthropogenic impacts (habitat degradation, climate change, and biological invasions, among others) with a high number of endemic range-restricted freshwater species. Despite the urgency, current conservation actions fall short of providing adequate protection. The irreplaceability index has been proposed as a useful assessment tool to focus limited efforts on areas that provide the highest benefit for threatened species. However, the transboundary nature of many rivers in the Iberian Peninsula can be a source of inefficiencies in protection if prioritisation efforts are conducted at a national rather than a peninsular scale. Objective: The aim of this study is to identify priority conservation basins for threatened native freshwater fish in the Iberian Peninsula and to evaluate the impact of national versus transboundary management strategies on the spatial protection afforded to these species. Methodology: The irreplaceability index was calculated for each basin by integrating basin richness, species rarity and their IUCN Red List conservation status. First, we modelled the species’ probability of presence using field observations recorded since 2000. Rarity was then calculated as the ratio between the modelled probability and the total number of basins within the species’ theoretical natural distribution. We then weighted each species’ rarity by its IUCN Red List conservation status, with higher weights to threatened species. We then calculated the basin irreplaceability index as the sum across all the species present in the basin of their conservation status-weighted rarity and ranked them according to this index. We replicated this approach considering Spain and Portugal independently, and both countries as one conservation planning unit. Results and Conclusions: The most irreplaceable basins were those harbouring a high density of threatened, narrow-range endemics. The priorities in each country differ depending on whether management strategies adopt a national or a broader geographical approach. Therefore, effective conservation requires transboundary planification to safeguard the shared biodiversity across countries. 2026-06-22 Proceedings, Vol. 146, Pages 87: Identifying Priority Conservation Areas for Iberian Freshwater Fish: National vs. Transboundary Approach

Proceedings doi: 10.3390/proceedings2026146087

Authors: Ignacio Pons Imanol Miqueleiz Marta Rodríguez Rey Rafael Miranda

Introduction: Freshwater habitats underpin global biodiversity and provide an array of essential ecosystem services to humans. However, threat hotspots like the Iberian Peninsula combine severe anthropogenic impacts (habitat degradation, climate change, and biological invasions, among others) with a high number of endemic range-restricted freshwater species. Despite the urgency, current conservation actions fall short of providing adequate protection. The irreplaceability index has been proposed as a useful assessment tool to focus limited efforts on areas that provide the highest benefit for threatened species. However, the transboundary nature of many rivers in the Iberian Peninsula can be a source of inefficiencies in protection if prioritisation efforts are conducted at a national rather than a peninsular scale. Objective: The aim of this study is to identify priority conservation basins for threatened native freshwater fish in the Iberian Peninsula and to evaluate the impact of national versus transboundary management strategies on the spatial protection afforded to these species. Methodology: The irreplaceability index was calculated for each basin by integrating basin richness, species rarity and their IUCN Red List conservation status. First, we modelled the species’ probability of presence using field observations recorded since 2000. Rarity was then calculated as the ratio between the modelled probability and the total number of basins within the species’ theoretical natural distribution. We then weighted each species’ rarity by its IUCN Red List conservation status, with higher weights to threatened species. We then calculated the basin irreplaceability index as the sum across all the species present in the basin of their conservation status-weighted rarity and ranked them according to this index. We replicated this approach considering Spain and Portugal independently, and both countries as one conservation planning unit. Results and Conclusions: The most irreplaceable basins were those harbouring a high density of threatened, narrow-range endemics. The priorities in each country differ depending on whether management strategies adopt a national or a broader geographical approach. Therefore, effective conservation requires transboundary planification to safeguard the shared biodiversity across countries.

]]> Identifying Priority Conservation Areas for Iberian Freshwater Fish: National vs. Transboundary Approach Ignacio Pons Imanol Miqueleiz Marta Rodríguez Rey Rafael Miranda doi: 10.3390/proceedings2026146087 Proceedings 2026-06-22 Proceedings 2026-06-22 146 1 Abstract 87 10.3390/proceedings2026146087 https://www.mdpi.com/2504-3900/146/1/87 Proceedings, Vol. 146, Pages 86: Hybridization as an Emerging Threat to Iberian Freshwater Ichthyofauna https://www.mdpi.com/2504-3900/146/1/86 In the Iberian Peninsula, the rapid expansion of the invasive species Alburnus alburnus (bleak) has intensified its contact with several endemic cyprinid species, raising concerns about hybridization, introgression, and the loss of genetic diversity. Despite increasing evidence of hybridization, data remain limited for many Iberian River basins, where endemic species persist in fragmented and vulnerable habitats. The aim of this study is to assess the extent and spatial distribution of hybridization between the bleak and the following native cyprinid species, Anaecypris hispanica (jarabugo), Iberochondrostoma lemmingii (pardilla), Pseudochondrostoma willkommii (Guadiana nase), and Squalius alburnoides (calandino), across several rivers within the Guadalquivir Basin. To this end, Sanger sequencing will be performed on the mitochondrial cytochrome b gene (maternal lineage; approximately 1000 base pairs (bp)) and the nuclear β-actin gene (paternal lineage; approximately 950 bp) from individuals of all endemic species and the bleak. Parental species and putative hybrids were initially identified in the field using diagnostic morphological and meristic characters, including number of rays in the anal fin morphology, mouth position, and the number of lateral line scales. Molecular analyses will include haplotype network reconstruction and phylogenetic tree inference to evaluate relationships among individuals from different species and to assess lineage divergence. The results will allow us to: (1) detect hybrids between the bleak and endemic cyprinids, (2) identify hybridization events among endemic cyprinid species, and (3) evaluate the correspondence between diagnostic morphological and meristic traits and the molecular identification of hybrid individuals. Overall, these findings will provide key information for the conservation management of endangered Iberian freshwater fishes in the context of invasive species expansion and global change. 2026-06-22 Proceedings, Vol. 146, Pages 86: Hybridization as an Emerging Threat to Iberian Freshwater Ichthyofauna

Proceedings doi: 10.3390/proceedings2026146086

Authors: Álvaro Checa Felipe Morcillo Paloma Alcorlo Anabel Perdices

In the Iberian Peninsula, the rapid expansion of the invasive species Alburnus alburnus (bleak) has intensified its contact with several endemic cyprinid species, raising concerns about hybridization, introgression, and the loss of genetic diversity. Despite increasing evidence of hybridization, data remain limited for many Iberian River basins, where endemic species persist in fragmented and vulnerable habitats. The aim of this study is to assess the extent and spatial distribution of hybridization between the bleak and the following native cyprinid species, Anaecypris hispanica (jarabugo), Iberochondrostoma lemmingii (pardilla), Pseudochondrostoma willkommii (Guadiana nase), and Squalius alburnoides (calandino), across several rivers within the Guadalquivir Basin. To this end, Sanger sequencing will be performed on the mitochondrial cytochrome b gene (maternal lineage; approximately 1000 base pairs (bp)) and the nuclear β-actin gene (paternal lineage; approximately 950 bp) from individuals of all endemic species and the bleak. Parental species and putative hybrids were initially identified in the field using diagnostic morphological and meristic characters, including number of rays in the anal fin morphology, mouth position, and the number of lateral line scales. Molecular analyses will include haplotype network reconstruction and phylogenetic tree inference to evaluate relationships among individuals from different species and to assess lineage divergence. The results will allow us to: (1) detect hybrids between the bleak and endemic cyprinids, (2) identify hybridization events among endemic cyprinid species, and (3) evaluate the correspondence between diagnostic morphological and meristic traits and the molecular identification of hybrid individuals. Overall, these findings will provide key information for the conservation management of endangered Iberian freshwater fishes in the context of invasive species expansion and global change.

]]> Hybridization as an Emerging Threat to Iberian Freshwater Ichthyofauna Álvaro Checa Felipe Morcillo Paloma Alcorlo Anabel Perdices doi: 10.3390/proceedings2026146086 Proceedings 2026-06-22 Proceedings 2026-06-22 146 1 Abstract 86 10.3390/proceedings2026146086 https://www.mdpi.com/2504-3900/146/1/86 Proceedings, Vol. 146, Pages 81: Phylogeography of Thorichthys meeki in Southeastern Mexico (Actinopteri: Cichlidae) https://www.mdpi.com/2504-3900/146/1/81 Introduction: The fish fauna in the Yucatan Peninsula exhibit a unique composition, shaped by the region’s complex geological and hydrological history. Thorichthys meeki is a widely distributed cichlid species that has been the subject of morphological, behavioral, and phylogenetic studies. Nevertheless, the historical and biogeographical processes that have shaped its diversification and genetic structure remain unexplored. Objective: The objective of this study was to identify phylogeographic patterns and assess the genetic diversity of T. meeki in southeastern Mexico. Methodology: Genomic DNA was extracted from the muscle tissue of specimens preserved in 90% ethanol. DNA was amplified using a polymerase chain reaction (PCR) and sequences were obtained from two molecular markers: cytochrome oxidase 1 (CO1) and S7 intron 1. Phylogeographic analyses included genetic diversity indices molecular variance analyses to assess population structure and haplotype networks. Results: A total of 60 CO1 and 40 S7 intron 1 sequences were obtained from 26 sampling sites. For CO1, 14 haplotypes were identified, with high haplotype diversity (Hd = 0.84), and low nucleotide diversity (π = 0.0048). For S7 intron 1, 16 haplotypes were recovered with lower haplotype diversity (Hd = 0.543) and π = 0.021. A hierarchical AMOVA (2 groups) showed the greatest variation; 72.3% in CO1 and 79.8% in S7 within population groups with significant ΦST values. Haplotype networks identified three haplogroups for each gene. A single CO1 haplotype was found in 26 sequences from 13 sampling sites. For S7 intron 1, one haplotype was identified in 22 sequences and was shered from all populations. Conclusions: A widely distributed haplotype for CO1 and S7 reflects historical connectivity among hydrological systems. These preliminary results provide an interesting insights into the processesthat have influenced the distribution of T. meeki on the Yucatán Peninsula. 2026-06-22 Proceedings, Vol. 146, Pages 81: Phylogeography of Thorichthys meeki in Southeastern Mexico (Actinopteri: Cichlidae)

Proceedings doi: 10.3390/proceedings2026146081

Authors: Eduardo López-Segovia Luis Fernando Del Moral-Flores Juan J. Schmitter-Soto Eduardo López-Urrutia Verónica García-Castillo Carlos Pérez-Plasencia Omar Domínguez-Domínguez Eloísa Torres-Hernández Jorge Ciros-Pérez

Introduction: The fish fauna in the Yucatan Peninsula exhibit a unique composition, shaped by the region’s complex geological and hydrological history. Thorichthys meeki is a widely distributed cichlid species that has been the subject of morphological, behavioral, and phylogenetic studies. Nevertheless, the historical and biogeographical processes that have shaped its diversification and genetic structure remain unexplored. Objective: The objective of this study was to identify phylogeographic patterns and assess the genetic diversity of T. meeki in southeastern Mexico. Methodology: Genomic DNA was extracted from the muscle tissue of specimens preserved in 90% ethanol. DNA was amplified using a polymerase chain reaction (PCR) and sequences were obtained from two molecular markers: cytochrome oxidase 1 (CO1) and S7 intron 1. Phylogeographic analyses included genetic diversity indices molecular variance analyses to assess population structure and haplotype networks. Results: A total of 60 CO1 and 40 S7 intron 1 sequences were obtained from 26 sampling sites. For CO1, 14 haplotypes were identified, with high haplotype diversity (Hd = 0.84), and low nucleotide diversity (π = 0.0048). For S7 intron 1, 16 haplotypes were recovered with lower haplotype diversity (Hd = 0.543) and π = 0.021. A hierarchical AMOVA (2 groups) showed the greatest variation; 72.3% in CO1 and 79.8% in S7 within population groups with significant ΦST values. Haplotype networks identified three haplogroups for each gene. A single CO1 haplotype was found in 26 sequences from 13 sampling sites. For S7 intron 1, one haplotype was identified in 22 sequences and was shered from all populations. Conclusions: A widely distributed haplotype for CO1 and S7 reflects historical connectivity among hydrological systems. These preliminary results provide an interesting insights into the processesthat have influenced the distribution of T. meeki on the Yucatán Peninsula.

]]> Phylogeography of Thorichthys meeki in Southeastern Mexico (Actinopteri: Cichlidae) Eduardo López-Segovia Luis Fernando Del Moral-Flores Juan J. Schmitter-Soto Eduardo López-Urrutia Verónica García-Castillo Carlos Pérez-Plasencia Omar Domínguez-Domínguez Eloísa Torres-Hernández Jorge Ciros-Pérez doi: 10.3390/proceedings2026146081 Proceedings 2026-06-22 Proceedings 2026-06-22 146 1 Abstract 81 10.3390/proceedings2026146081 https://www.mdpi.com/2504-3900/146/1/81 Proceedings, Vol. 146, Pages 80: Assessing Long-Term Drought Effects on Guadalquivir Estuary Nursery Function and Fisheries Production Based on a Long-Term Ecological Research Project: Guadalquivir_LTER 1997–2027 https://www.mdpi.com/2504-3900/146/1/80 Introduction: Climate change is reducing freshwater availability worldwide, making it essential to understand how freshwater inflow influences estuarine ecosystem functioning and marine fisheries productivity. In the Gulf of Cádiz (SW Spain), one of the most important fishing areas in Spain, the Guadalquivir Estuary serves as a key nursery habitat for commercially important fish and crustacean species. Objective: The aim of this study is to evaluate the effects of droughts and floods on estuarine functioning and coastal fisheries. Methodology: We analyzed 25 years of monthly data (1997–2022) from the Guadalquivir Long-Term Ecological Research Program (GUADALQUIVIR-LTER), using time-series analyses and dynamic structural equation modelling. Environmental variables, zooplankton and mysid biomass, and juvenile biomass of anchovy, sardine, and meagre were examined to assess trophic relationships and recruitment dynamics. Results: Our findings show that positive North Atlantic Oscillation (NAO) phases, associated with drought conditions in southern Europe, reduced freshwater inflow from the Alcalá del Río Dam into the estuary. Freshwater input increased organic matter and turbidity, which positively affected the mysid Rhopalophthalmus tartessicus, an important prey species for anchovy recruits. The mysid Mesopodopsis slabberi showed the strongest positive effect on anchovy recruitment (0.39). Although turbidity initially had a negative effect on M. slabberi, a significant positive effect appeared after monthly lag = 4. Conclusions: These findings demonstrate that spring freshwater inflow is essential for maintaining estuarine productivity, enhancing mysid abundance, and supporting anchovy recruitment, ultimately benefiting adult fish stocks after juveniles migrate from the estuary to coastal waters. 2026-06-22 Proceedings, Vol. 146, Pages 80: Assessing Long-Term Drought Effects on Guadalquivir Estuary Nursery Function and Fisheries Production Based on a Long-Term Ecological Research Project: Guadalquivir_LTER 1997–2027

Proceedings doi: 10.3390/proceedings2026146080

Authors: César Vilas Ray Czaja Arnaud Grüss Stefenia van Bergeijk Enrique González-Ortegón J. Pedro Cañavate

Introduction: Climate change is reducing freshwater availability worldwide, making it essential to understand how freshwater inflow influences estuarine ecosystem functioning and marine fisheries productivity. In the Gulf of Cádiz (SW Spain), one of the most important fishing areas in Spain, the Guadalquivir Estuary serves as a key nursery habitat for commercially important fish and crustacean species. Objective: The aim of this study is to evaluate the effects of droughts and floods on estuarine functioning and coastal fisheries. Methodology: We analyzed 25 years of monthly data (1997–2022) from the Guadalquivir Long-Term Ecological Research Program (GUADALQUIVIR-LTER), using time-series analyses and dynamic structural equation modelling. Environmental variables, zooplankton and mysid biomass, and juvenile biomass of anchovy, sardine, and meagre were examined to assess trophic relationships and recruitment dynamics. Results: Our findings show that positive North Atlantic Oscillation (NAO) phases, associated with drought conditions in southern Europe, reduced freshwater inflow from the Alcalá del Río Dam into the estuary. Freshwater input increased organic matter and turbidity, which positively affected the mysid Rhopalophthalmus tartessicus, an important prey species for anchovy recruits. The mysid Mesopodopsis slabberi showed the strongest positive effect on anchovy recruitment (0.39). Although turbidity initially had a negative effect on M. slabberi, a significant positive effect appeared after monthly lag = 4. Conclusions: These findings demonstrate that spring freshwater inflow is essential for maintaining estuarine productivity, enhancing mysid abundance, and supporting anchovy recruitment, ultimately benefiting adult fish stocks after juveniles migrate from the estuary to coastal waters.

]]> Assessing Long-Term Drought Effects on Guadalquivir Estuary Nursery Function and Fisheries Production Based on a Long-Term Ecological Research Project: Guadalquivir_LTER 1997–2027 César Vilas Ray Czaja Arnaud Grüss Stefenia van Bergeijk Enrique González-Ortegón J. Pedro Cañavate doi: 10.3390/proceedings2026146080 Proceedings 2026-06-22 Proceedings 2026-06-22 146 1 Abstract 80 10.3390/proceedings2026146080 https://www.mdpi.com/2504-3900/146/1/80 Proceedings, Vol. 146, Pages 79: Trends in Conservation and Exploitation of Skates (Rajidae) in the Northeast Atlantic and Mediterranean: Implications for Management https://www.mdpi.com/2504-3900/146/1/79 Introduction: Skates (Rajidae) are cornerstone elasmobranchs, yet their intrinsic biological constraints, like slow growth, late maturation, and low fecundity, render them exceptionally susceptible to anthropogenic pressure. Despite their ecological and economic importance, tracking their population trajectories is historically hindered by “taxonomic blurring” and aggregated reporting in commercial fisheries. Objective: This study evaluates long-term conservation trends and exploitation dynamics of Rajidae species in the Northeast Atlantic and the Mediterranean Sea. Methodology: We analyzed 31 Rajidae species across the Northeast Atlantic and the Mediterranean Sea (FAO Areas 27 and 37) by integrating IUCN Red List assessments, species-specific life-history traits (maximum body size and depth distribution), and FAO fisheries landing data from 1992 to 2023. Descriptive analyses and Spearman correlations were used to assess temporal trends in conservation status and exploitation patterns. Results: Our synthesis reveals that some species show improvements in IUCN Red List category assessments, likely driven by recent management interventions such as species-specific reporting, catch quotas, and targeted retention bans. However, we also identify a critical mismatch between policy and biology: current Total Allowable Catches (TACs) and minimum landing sizes often do not explicitly incorporate species-specific life-history traits, inadvertently favoring smaller, less-marketable taxa while leaving larger, vulnerable species at risk. While FAO landings offer a valuable broad-scale overview of exploitation, the results highlight the limitations of aggregated fisheries statistics for species-level conservation assessments. Conclusions: These findings underline the need to adopt more precise and species-specific fisheries management approaches for Rajidae, including expanded regional monitoring programs, the use of data collected by on-board observers or electronic monitoring tools, and improved control of data reporting procedures, to prevent continued aggregation of species-level data. 2026-06-19 Proceedings, Vol. 146, Pages 79: Trends in Conservation and Exploitation of Skates (Rajidae) in the Northeast Atlantic and Mediterranean: Implications for Management

Proceedings doi: 10.3390/proceedings2026146079

Authors: Sara Lourenço Catarina N. S. Silva Miguel A. Pardal Paolo Momigliano André S. Afonso Filipe Martinho

Introduction: Skates (Rajidae) are cornerstone elasmobranchs, yet their intrinsic biological constraints, like slow growth, late maturation, and low fecundity, render them exceptionally susceptible to anthropogenic pressure. Despite their ecological and economic importance, tracking their population trajectories is historically hindered by “taxonomic blurring” and aggregated reporting in commercial fisheries. Objective: This study evaluates long-term conservation trends and exploitation dynamics of Rajidae species in the Northeast Atlantic and the Mediterranean Sea. Methodology: We analyzed 31 Rajidae species across the Northeast Atlantic and the Mediterranean Sea (FAO Areas 27 and 37) by integrating IUCN Red List assessments, species-specific life-history traits (maximum body size and depth distribution), and FAO fisheries landing data from 1992 to 2023. Descriptive analyses and Spearman correlations were used to assess temporal trends in conservation status and exploitation patterns. Results: Our synthesis reveals that some species show improvements in IUCN Red List category assessments, likely driven by recent management interventions such as species-specific reporting, catch quotas, and targeted retention bans. However, we also identify a critical mismatch between policy and biology: current Total Allowable Catches (TACs) and minimum landing sizes often do not explicitly incorporate species-specific life-history traits, inadvertently favoring smaller, less-marketable taxa while leaving larger, vulnerable species at risk. While FAO landings offer a valuable broad-scale overview of exploitation, the results highlight the limitations of aggregated fisheries statistics for species-level conservation assessments. Conclusions: These findings underline the need to adopt more precise and species-specific fisheries management approaches for Rajidae, including expanded regional monitoring programs, the use of data collected by on-board observers or electronic monitoring tools, and improved control of data reporting procedures, to prevent continued aggregation of species-level data.

]]> Trends in Conservation and Exploitation of Skates (Rajidae) in the Northeast Atlantic and Mediterranean: Implications for Management Sara Lourenço Catarina N. S. Silva Miguel A. Pardal Paolo Momigliano André S. Afonso Filipe Martinho doi: 10.3390/proceedings2026146079 Proceedings 2026-06-19 Proceedings 2026-06-19 146 1 Abstract 79 10.3390/proceedings2026146079 https://www.mdpi.com/2504-3900/146/1/79 Proceedings, Vol. 146, Pages 77: River Restoration Through Weir Removal Enhances Native Fish Connectivity in a Mediterranean River https://www.mdpi.com/2504-3900/146/1/77 Introduction: The Guadiana basin harbours one of the most threatened freshwater fish faunas in Europe, concentrating a high number of Iberian endemisms and species of high conservation interest. Among these, the Iberian minnowcarp (Anaecypris hispanica) and the Iberian Arched-Mouth Nase (Iberochondrostoma lemmingii) stand out as threatened species whose populations are in serious decline. Methodology: In March 2023, an obsolete river barrier, the Galaches weir, was removed from the Odeleite River (Guadiana basin), releasing an 8 km lotic stretch previously inaccessible to downstream fish fauna. This study aimed to evaluate the effects of weir removal on the composition and structure of the fish community, using a Before-After design with fish sampling conducted before (2022) and after (2024) weir removal. Fish sampling took place in early summer by means of electrofishing in two 150 m long river sites, one upstream and one downstream of the weir. Results: Before removal, both upstream and downstream sites were dominated by native leuciscids (A. hispanica, I. lemmingii, Squalius pyrenaicus, and Pseudochondrostoma willkommii) and cyprinids (Luciobarbus spp.), with non-natives Lepomis gibbosus and Australoheros facetus occurring in lower abundance. After removal, native species remained clearly dominant at both sites, while A. facetus was no longer detected and L. gibbosus did not increase in abundance, suggesting that barrier removal did not favour the spread of non-native fish. Species size–structure revealed the presence of multiple size classes, possibly supporting a regular recruitment of key native taxa: A. hispanica occurred almost exclusively with total lengths of 4 to 6 cm, whereas I. lemmingii and other native leuciscids displayed broader size ranges, spanning juveniles to adults, consistent with sustained reproduction and growth in the restored river stretch. Conclusions: These findings support the hypothesis that weir removal has improved the sustainability of native fish populations such as A. hispanica by improving habitat connectivity and increasing recruitment. The results highlight the importance of weir removal for the conservation of threatened species in Mediterranean river systems. 2026-06-19 Proceedings, Vol. 146, Pages 77: River Restoration Through Weir Removal Enhances Native Fish Connectivity in a Mediterranean River

Proceedings doi: 10.3390/proceedings2026146077

Authors: José Maria Santos Paulo Branco Maria João Costa Ana Filipa Filipe

Introduction: The Guadiana basin harbours one of the most threatened freshwater fish faunas in Europe, concentrating a high number of Iberian endemisms and species of high conservation interest. Among these, the Iberian minnowcarp (Anaecypris hispanica) and the Iberian Arched-Mouth Nase (Iberochondrostoma lemmingii) stand out as threatened species whose populations are in serious decline. Methodology: In March 2023, an obsolete river barrier, the Galaches weir, was removed from the Odeleite River (Guadiana basin), releasing an 8 km lotic stretch previously inaccessible to downstream fish fauna. This study aimed to evaluate the effects of weir removal on the composition and structure of the fish community, using a Before-After design with fish sampling conducted before (2022) and after (2024) weir removal. Fish sampling took place in early summer by means of electrofishing in two 150 m long river sites, one upstream and one downstream of the weir. Results: Before removal, both upstream and downstream sites were dominated by native leuciscids (A. hispanica, I. lemmingii, Squalius pyrenaicus, and Pseudochondrostoma willkommii) and cyprinids (Luciobarbus spp.), with non-natives Lepomis gibbosus and Australoheros facetus occurring in lower abundance. After removal, native species remained clearly dominant at both sites, while A. facetus was no longer detected and L. gibbosus did not increase in abundance, suggesting that barrier removal did not favour the spread of non-native fish. Species size–structure revealed the presence of multiple size classes, possibly supporting a regular recruitment of key native taxa: A. hispanica occurred almost exclusively with total lengths of 4 to 6 cm, whereas I. lemmingii and other native leuciscids displayed broader size ranges, spanning juveniles to adults, consistent with sustained reproduction and growth in the restored river stretch. Conclusions: These findings support the hypothesis that weir removal has improved the sustainability of native fish populations such as A. hispanica by improving habitat connectivity and increasing recruitment. The results highlight the importance of weir removal for the conservation of threatened species in Mediterranean river systems.

]]> River Restoration Through Weir Removal Enhances Native Fish Connectivity in a Mediterranean River José Maria Santos Paulo Branco Maria João Costa Ana Filipa Filipe doi: 10.3390/proceedings2026146077 Proceedings 2026-06-19 Proceedings 2026-06-19 146 1 Abstract 77 10.3390/proceedings2026146077 https://www.mdpi.com/2504-3900/146/1/77 Proceedings, Vol. 146, Pages 75: Impact of Small Weirs on the Distribution, Abundance, and Habitat Use of the Threatened Species Achondrostoma asturicense https://www.mdpi.com/2504-3900/146/1/75 Mediterranean rivers experience various pressures that cause native fish populations to decline. This is the case of Achondrostoma asturicense, a threatened endemic species recently classified as "endangered" (EN) by the Portuguese Red Book. In northeastern Portugal, the main populations occur in the Carvalhais (Tua basin), Maçãs, and Angueira (Sabor basin) rivers. This study aimed to evaluate the effect of small weirs on the distribution, abundance, and habitat use of A. asturicense populations along the longitudinal gradient of River Angueira. Three river zones (Z1 to Z3) affected by small weirs were selected along the watercourse. In each zone, four sampling sites were distributed relative to the weir, as follows: A1—200 m downstream; A2—immediately downstream; A3—under the influence of the reservoir; and A4—200 m upstream (reference). Sampling was carried out in three seasons: winter, spring, and summer 2025. Physical–chemical (water), hydromorphological (habitats), and biological (fish) elements were assessed in accordance with the protocols of the Water Framework Directive. The results revealed the worst water quality in the summer season due to flow reduction, dissolved oxygen depletion, elevated conductivity, and high nutrient (N, P) concentrations, leading to eutrophication phenomena. The distribution and abundance of A. asturicense were negatively influenced by the presence of small weirs, which were dominated by the non-native species Lepomis gibbosus. PERMANOVA analysis (two-way, p < 0.05) identified significant differences (Pseudo-F = 28.349, p < 0.05) between sampling sites and river zones, and paired similarity analysis tests (ANOSIM; one-way, p < 0.05) confirmed that these differences occur only between the weir reservoir (A3) and the remaining sampling sites. A. asturicense showed a maximum length (Lmax) of 135 mm, isometric growth only in the upstream zone (Z1), and significant differences in body condition (0.75 < K < 0.84). This species showed a preference for rheophilic habitats, with coarse substrate and diversity of currents and distinct cover mainly guaranteed by submerged aquatic macrophytes. A. asturicense populations are severely affected in the River Angueira by habitat fragmentation, riparian degradation, pollution, invasive alien species (e.g., L. gibbosus, P. clarkii, N. vison), and climate change, justifying the development of mitigation and restoration measures for the conservation of natural habitats and native threatened species. 2026-06-19 Proceedings, Vol. 146, Pages 75: Impact of Small Weirs on the Distribution, Abundance, and Habitat Use of the Threatened Species Achondrostoma asturicense

Proceedings doi: 10.3390/proceedings2026146075

Authors: Amílcar Teixeira Ryan Alves da Silva Douglas Santos Fernando Miranda Fernando Teixeira

Mediterranean rivers experience various pressures that cause native fish populations to decline. This is the case of Achondrostoma asturicense, a threatened endemic species recently classified as "endangered" (EN) by the Portuguese Red Book. In northeastern Portugal, the main populations occur in the Carvalhais (Tua basin), Maçãs, and Angueira (Sabor basin) rivers. This study aimed to evaluate the effect of small weirs on the distribution, abundance, and habitat use of A. asturicense populations along the longitudinal gradient of River Angueira. Three river zones (Z1 to Z3) affected by small weirs were selected along the watercourse. In each zone, four sampling sites were distributed relative to the weir, as follows: A1—200 m downstream; A2—immediately downstream; A3—under the influence of the reservoir; and A4—200 m upstream (reference). Sampling was carried out in three seasons: winter, spring, and summer 2025. Physical–chemical (water), hydromorphological (habitats), and biological (fish) elements were assessed in accordance with the protocols of the Water Framework Directive. The results revealed the worst water quality in the summer season due to flow reduction, dissolved oxygen depletion, elevated conductivity, and high nutrient (N, P) concentrations, leading to eutrophication phenomena. The distribution and abundance of A. asturicense were negatively influenced by the presence of small weirs, which were dominated by the non-native species Lepomis gibbosus. PERMANOVA analysis (two-way, p < 0.05) identified significant differences (Pseudo-F = 28.349, p < 0.05) between sampling sites and river zones, and paired similarity analysis tests (ANOSIM; one-way, p < 0.05) confirmed that these differences occur only between the weir reservoir (A3) and the remaining sampling sites. A. asturicense showed a maximum length (Lmax) of 135 mm, isometric growth only in the upstream zone (Z1), and significant differences in body condition (0.75 < K < 0.84). This species showed a preference for rheophilic habitats, with coarse substrate and diversity of currents and distinct cover mainly guaranteed by submerged aquatic macrophytes. A. asturicense populations are severely affected in the River Angueira by habitat fragmentation, riparian degradation, pollution, invasive alien species (e.g., L. gibbosus, P. clarkii, N. vison), and climate change, justifying the development of mitigation and restoration measures for the conservation of natural habitats and native threatened species.

]]> Impact of Small Weirs on the Distribution, Abundance, and Habitat Use of the Threatened Species Achondrostoma asturicense Amílcar Teixeira Ryan Alves da Silva Douglas Santos Fernando Miranda Fernando Teixeira doi: 10.3390/proceedings2026146075 Proceedings 2026-06-19 Proceedings 2026-06-19 146 1 Abstract 75 10.3390/proceedings2026146075 https://www.mdpi.com/2504-3900/146/1/75 Proceedings, Vol. 146, Pages 101: Outreach Programme LIFE PREDATOR: From Schools to Fishermen https://www.mdpi.com/2504-3900/146/1/101 Introduction: Knowledge of Iberian freshwater fish fauna among the general public is scarce and generally limited to a handful of species. Moreover, this knowledge gap increases as time goes by, particularly in younger generations, due to the lack of content on native fish fauna in school programmes. Nevertheless, schools across the country are proving increasingly receptive to innovative approaches that engage students in meaningful, real-world learning. Objectives: The LIFE PREDATOR programme leverages this opportunity to educate young people about freshwater fish communities. It aims to prevent the spread of the largest invasive fish in Portugal, the European catfish (Silurus glanis), by engaging students as active conservation ambassadors. Methodology: In inland Portugal, fishing is a cultural practice, and children frequently participate in angling activities alongside friends and family members. By reaching children, the programme simultaneously targets future anglers, potential decision-makers, and a channel for intergenerational knowledge transfer. Results: Thus far, the programme has reached over 5000 students and almost 60 schools, mostly throughout the Tagus basin. Preliminary assessments revealed improvements in students’ ability to name emblematic native fish species like the Iberian nase (Pseudochondrostoma spp.), European eel (Anguilla anguilla) and Iberian barbel (Luciobarbus spp.), and recognise the threats posed by invasives like the European catfish. To ensure national scalability, we have developed learning materials designed for use by teachers across Portugal, which are to be made available for free online. Beyond the dissemination directed to adult fishermen, which is often more demanding, Conclusions: LIFE PREDATOR ensures that knowledge about native river fauna, invasive species, and responsible fishing practices is conveyed through trusted, familiar voices. This intergenerational transmission model has the potential to embed long-lasting behavioural change within future fishing communities. 2026-06-18 Proceedings, Vol. 146, Pages 101: Outreach Programme LIFE PREDATOR: From Schools to Fishermen

Proceedings doi: 10.3390/proceedings2026146101

Authors: Mafalda Moncada Diogo Ribeiro Beatriz Castro Diogo Dias Rui Rivaes Filipe Ribeiro

Introduction: Knowledge of Iberian freshwater fish fauna among the general public is scarce and generally limited to a handful of species. Moreover, this knowledge gap increases as time goes by, particularly in younger generations, due to the lack of content on native fish fauna in school programmes. Nevertheless, schools across the country are proving increasingly receptive to innovative approaches that engage students in meaningful, real-world learning. Objectives: The LIFE PREDATOR programme leverages this opportunity to educate young people about freshwater fish communities. It aims to prevent the spread of the largest invasive fish in Portugal, the European catfish (Silurus glanis), by engaging students as active conservation ambassadors. Methodology: In inland Portugal, fishing is a cultural practice, and children frequently participate in angling activities alongside friends and family members. By reaching children, the programme simultaneously targets future anglers, potential decision-makers, and a channel for intergenerational knowledge transfer. Results: Thus far, the programme has reached over 5000 students and almost 60 schools, mostly throughout the Tagus basin. Preliminary assessments revealed improvements in students’ ability to name emblematic native fish species like the Iberian nase (Pseudochondrostoma spp.), European eel (Anguilla anguilla) and Iberian barbel (Luciobarbus spp.), and recognise the threats posed by invasives like the European catfish. To ensure national scalability, we have developed learning materials designed for use by teachers across Portugal, which are to be made available for free online. Beyond the dissemination directed to adult fishermen, which is often more demanding, Conclusions: LIFE PREDATOR ensures that knowledge about native river fauna, invasive species, and responsible fishing practices is conveyed through trusted, familiar voices. This intergenerational transmission model has the potential to embed long-lasting behavioural change within future fishing communities.

]]> Outreach Programme LIFE PREDATOR: From Schools to Fishermen Mafalda Moncada Diogo Ribeiro Beatriz Castro Diogo Dias Rui Rivaes Filipe Ribeiro doi: 10.3390/proceedings2026146101 Proceedings 2026-06-18 Proceedings 2026-06-18 146 1 Abstract 101 10.3390/proceedings2026146101 https://www.mdpi.com/2504-3900/146/1/101 Proceedings, Vol. 146, Pages 100: Movements and Dispersal of Wild and Stocked Brown Trout (Salmo trutta Linnaeus, 1758) in Mountain Rivers of NE Portugal https://www.mdpi.com/2504-3900/146/1/100 Brown trout (Salmo trutta L.) is a bioindicator species of ecological integrity in mountain rivers of northern Portugal. Habitat loss and recreational fishing justify sustainable management to balance the conservation and exploitation of these fish populations. In fact, salmonid streams in NE Portugal are low productive watercourses and fish stocking has been continuously demanded by fishermen. However, this most common management action must be analyzed carefully and determined the effective increase for local fisheries, taking into consideration the potential dispersal of stocked fish. The objective of the present study, developed in River Sabor, was to determine short- and medium-term movement and dispersal patterns and habitat preferences of wild and stocked Brown trout, using radio telemetry, during a weekly monitoring 4-month period (October to February 2026; n = 18). Fish was sampled by electrofishing at beginning and the end of the experiment. Twenty-four adult Brown trout, equally distributed by two salmonid sections, and three groups, (1) wild resident (River Sabor) (213–270 mm TL); (2) wild non-resident (from contiguous basin, River Baceiro) (200–375 mm TL) and (3) rear-captivity (Castrelos Fishfarms, ICNF) (227–365 mm TL) fish, were surgically implanted with radio transmitters. Significant differences (KW-H (2;24) = 4.67; p = 0.09) were observed for the dispersal distances, considering fish detected at least in five sampling events, ranging from 120–1437 m for the wild resident stationary group to 192–14,150 m for the stocked mobile group. Moreover, wild non-resident fish displayed higher movement in the upstream direction, in opposition to the downstream movement of stocked individuals. Wild resident and non-resident trout tended to display increased movements during November and December, probably related to spawning activity, showing preferences by riffle and run habitats. Stocked fish were detected in pool habitats (mainly weir reservoirs), exhibiting significantly lower growth rates, and increased movement during January and February, particularly during flood events. These findings are valuable information for managers related to movement patterns, habitat use and stocking management. 2026-06-18 Proceedings, Vol. 146, Pages 100: Movements and Dispersal of Wild and Stocked Brown Trout (Salmo trutta Linnaeus, 1758) in Mountain Rivers of NE Portugal

Proceedings doi: 10.3390/proceedings2026146100

Authors: Amílcar Teixeira Fernando Miranda Fernando Teixeira

Brown trout (Salmo trutta L.) is a bioindicator species of ecological integrity in mountain rivers of northern Portugal. Habitat loss and recreational fishing justify sustainable management to balance the conservation and exploitation of these fish populations. In fact, salmonid streams in NE Portugal are low productive watercourses and fish stocking has been continuously demanded by fishermen. However, this most common management action must be analyzed carefully and determined the effective increase for local fisheries, taking into consideration the potential dispersal of stocked fish. The objective of the present study, developed in River Sabor, was to determine short- and medium-term movement and dispersal patterns and habitat preferences of wild and stocked Brown trout, using radio telemetry, during a weekly monitoring 4-month period (October to February 2026; n = 18). Fish was sampled by electrofishing at beginning and the end of the experiment. Twenty-four adult Brown trout, equally distributed by two salmonid sections, and three groups, (1) wild resident (River Sabor) (213–270 mm TL); (2) wild non-resident (from contiguous basin, River Baceiro) (200–375 mm TL) and (3) rear-captivity (Castrelos Fishfarms, ICNF) (227–365 mm TL) fish, were surgically implanted with radio transmitters. Significant differences (KW-H (2;24) = 4.67; p = 0.09) were observed for the dispersal distances, considering fish detected at least in five sampling events, ranging from 120–1437 m for the wild resident stationary group to 192–14,150 m for the stocked mobile group. Moreover, wild non-resident fish displayed higher movement in the upstream direction, in opposition to the downstream movement of stocked individuals. Wild resident and non-resident trout tended to display increased movements during November and December, probably related to spawning activity, showing preferences by riffle and run habitats. Stocked fish were detected in pool habitats (mainly weir reservoirs), exhibiting significantly lower growth rates, and increased movement during January and February, particularly during flood events. These findings are valuable information for managers related to movement patterns, habitat use and stocking management.

]]> Movements and Dispersal of Wild and Stocked Brown Trout (Salmo trutta Linnaeus, 1758) in Mountain Rivers of NE Portugal Amílcar Teixeira Fernando Miranda Fernando Teixeira doi: 10.3390/proceedings2026146100 Proceedings 2026-06-18 Proceedings 2026-06-18 146 1 Abstract 100 10.3390/proceedings2026146100 https://www.mdpi.com/2504-3900/146/1/100 Proceedings, Vol. 146, Pages 78: First Evidence of Gene Duplications Related with Hyaluronic Acid Metabolism in Elopomorpha https://www.mdpi.com/2504-3900/146/1/78 Elopomorpha is one of the most primitive teleost lineages and is characterised by a unique trait, the leptocephalus larva. Its leaf-like and transparent body is largely composed of glycosaminoglycan (GAG), which makes it distinctive from other fish larvae. Hyaluronic acid (HA) has been identified as the principal GAG present in these larvae and is thought to play several physiological roles, such as floatability, osmoregulation, and energy storage. During the larval stage, HA reaches a concentration of up to 40 to 50% of the body composition due to a HA synthesis peak. Later in metamorphosis, HA concentration decreases and, after this, adults show a normal HA metabolism. These observations could be explained if the larva has an alternative HA metabolism compared to the adult. It is known that HA synthesis is done by three hyaluronan synthase (HAS) enzymes, which differ in the length of HA synthetized, codified by three genes: HAS1, HAS2 and HAS3. Therefore, to test the hypothesis of a larval-specific metabolism, the genomic basis of these genes in Elopomorpha will be analysed. To this end, HAS coding and protein sequences from nine Elopomorpha, four Osteoglossomorpha and 21 Clupeocephala species were mined from the NCBI database to ensure a broad phylogenetic coverage. To guarantee a complete retrieval of HAS genes, the BLAST tool (https://blast.ncbi.nlm.nih.gov/Blast.cgi, accessed on 1 March 2026) and synteny analysis were performed. Then, Pseudochecker was used to confirm the functionality of the candidate genes obtained. Functional genes were used for phylogenetic analyses, maximum likelihood and Bayesian inference to test orthology. Both lineage and species duplications of HAS1 and HAS2 were found in Elopomorpha, which would be responsible for the synthesis of HA during the larval stage. These new genetic patterns provide the first evidence of a possible alternative metabolism of HA during the larval stage of Elopomorpha. 2026-06-18 Proceedings, Vol. 146, Pages 78: First Evidence of Gene Duplications Related with Hyaluronic Acid Metabolism in Elopomorpha

Proceedings doi: 10.3390/proceedings2026146078

Authors: Zoé Lemoine André Gomes-dos-Santos David Barros-García

Elopomorpha is one of the most primitive teleost lineages and is characterised by a unique trait, the leptocephalus larva. Its leaf-like and transparent body is largely composed of glycosaminoglycan (GAG), which makes it distinctive from other fish larvae. Hyaluronic acid (HA) has been identified as the principal GAG present in these larvae and is thought to play several physiological roles, such as floatability, osmoregulation, and energy storage. During the larval stage, HA reaches a concentration of up to 40 to 50% of the body composition due to a HA synthesis peak. Later in metamorphosis, HA concentration decreases and, after this, adults show a normal HA metabolism. These observations could be explained if the larva has an alternative HA metabolism compared to the adult. It is known that HA synthesis is done by three hyaluronan synthase (HAS) enzymes, which differ in the length of HA synthetized, codified by three genes: HAS1, HAS2 and HAS3. Therefore, to test the hypothesis of a larval-specific metabolism, the genomic basis of these genes in Elopomorpha will be analysed. To this end, HAS coding and protein sequences from nine Elopomorpha, four Osteoglossomorpha and 21 Clupeocephala species were mined from the NCBI database to ensure a broad phylogenetic coverage. To guarantee a complete retrieval of HAS genes, the BLAST tool (https://blast.ncbi.nlm.nih.gov/Blast.cgi, accessed on 1 March 2026) and synteny analysis were performed. Then, Pseudochecker was used to confirm the functionality of the candidate genes obtained. Functional genes were used for phylogenetic analyses, maximum likelihood and Bayesian inference to test orthology. Both lineage and species duplications of HAS1 and HAS2 were found in Elopomorpha, which would be responsible for the synthesis of HA during the larval stage. These new genetic patterns provide the first evidence of a possible alternative metabolism of HA during the larval stage of Elopomorpha.

]]> First Evidence of Gene Duplications Related with Hyaluronic Acid Metabolism in Elopomorpha Zoé Lemoine André Gomes-dos-Santos David Barros-García doi: 10.3390/proceedings2026146078 Proceedings 2026-06-18 Proceedings 2026-06-18 146 1 Abstract 78 10.3390/proceedings2026146078 https://www.mdpi.com/2504-3900/146/1/78 Proceedings, Vol. 146, Pages 76: Optimizing Fishway Efficiency Through an Integrated Adaptive Management Framework: A Case Study in the Duero River https://www.mdpi.com/2504-3900/146/1/76 Introduction: River fragmentation caused by hydropower infrastructure remains a primary threat to aquatic biodiversity, creating a critical need for fish passage solutions that can adapt to high environmental variability. Although adaptive management (AM) has the potential to significantly improve longitudinal connectivity and ecological resilience, its application in real-world fishway operations is currently limited. Objective: This study aims to present and validate a flexible AM framework designed to optimize fish passage by integrating low-cost monitoring systems with automated data processing and predictive modeling. Methodology: The proposed system combines a sensor network for real-time water-level and environmental monitoring with biological performance data obtained through Passive Integrated Transponder (PIT) technology. These data were processed locally using edge computing. Over a two-year period, weekly aggregated data were used to develop Random Forest models to identify the primary drivers of fish movement. Results: The final model successfully identified five key drivers: luminosity, water temperature, and three nested hydraulic parameters at the fishway’s upstream section. Validation at a vertical-slot fishway in Vadocondes (Duero River, Spain) showed that retrospective optimization—specifically adjusting sluice-gate regulation—could increase downstream water levels and reduce drops at the first cross wall. This adjustment demonstrated a substantial increase in predicted fish passage without requiring changes to the hydropower plant’s core operation. Conclusions: The framework is highly flexible and transferable to other regulated river systems. However, its success is contingent upon the definition of clear ecological objectives and the seamless integration of monitoring results into the day-to-day operation of river infrastructure. 2026-06-18 Proceedings, Vol. 146, Pages 76: Optimizing Fishway Efficiency Through an Integrated Adaptive Management Framework: A Case Study in the Duero River

Proceedings doi: 10.3390/proceedings2026146076

Authors: Marina Martínez-Miguel Ana García-Vega Francisco Javier Bravo-Córdoba Francisco J. Sanz-Ronda Juan Francisco Fuentes-Pérez

Introduction: River fragmentation caused by hydropower infrastructure remains a primary threat to aquatic biodiversity, creating a critical need for fish passage solutions that can adapt to high environmental variability. Although adaptive management (AM) has the potential to significantly improve longitudinal connectivity and ecological resilience, its application in real-world fishway operations is currently limited. Objective: This study aims to present and validate a flexible AM framework designed to optimize fish passage by integrating low-cost monitoring systems with automated data processing and predictive modeling. Methodology: The proposed system combines a sensor network for real-time water-level and environmental monitoring with biological performance data obtained through Passive Integrated Transponder (PIT) technology. These data were processed locally using edge computing. Over a two-year period, weekly aggregated data were used to develop Random Forest models to identify the primary drivers of fish movement. Results: The final model successfully identified five key drivers: luminosity, water temperature, and three nested hydraulic parameters at the fishway’s upstream section. Validation at a vertical-slot fishway in Vadocondes (Duero River, Spain) showed that retrospective optimization—specifically adjusting sluice-gate regulation—could increase downstream water levels and reduce drops at the first cross wall. This adjustment demonstrated a substantial increase in predicted fish passage without requiring changes to the hydropower plant’s core operation. Conclusions: The framework is highly flexible and transferable to other regulated river systems. However, its success is contingent upon the definition of clear ecological objectives and the seamless integration of monitoring results into the day-to-day operation of river infrastructure.

]]> Optimizing Fishway Efficiency Through an Integrated Adaptive Management Framework: A Case Study in the Duero River Marina Martínez-Miguel Ana García-Vega Francisco Javier Bravo-Córdoba Francisco J. Sanz-Ronda Juan Francisco Fuentes-Pérez doi: 10.3390/proceedings2026146076 Proceedings 2026-06-18 Proceedings 2026-06-18 146 1 Abstract 76 10.3390/proceedings2026146076 https://www.mdpi.com/2504-3900/146/1/76 Proceedings, Vol. 146, Pages 74: Widespread Gene Reorganizations in Teleost Mitochondria Are Driven by Ecological Transitions https://www.mdpi.com/2504-3900/146/1/74 The vertebrate mitochondrial genome (mitogenome) is a small, circular DNA molecule typically ~16–17 kb in length, encoding 37 genes that are essential for the electron transport chain, the mechanism that drives mostly all the ATP synthesis in cells. Owing to its central role in energy metabolism, its structure is highly conserved across vertebrate lineages in both the number and relative position of each gene in the genome. Nevertheless, different variations have been found in several teleost lineages, including antarctic fishes (Nototheniidae), gadiforms, hatchetfishes (Sternoptychidae), and Batrachoidiformes. The explanation for these phenomena remains unknown yet may reflect shifts in functional constraints and can provide insights into lineage-specific and/or coevolutionary processes. This raises the possibility that mitogenome structure is related to habitat selection, potentially reflecting environmental influences on energetic regulation. To further test this hypothesis, we studied more than 400 teleost species across all major teleost lineages. The mitogenome sequences were downloaded from NCBI and annotated using two independent algorithms (MITOZ and MITOS) and then compared with a reference (Danio rerio) to find any deviation from the standard structure. Similarly, ecological data was downloaded from FishBase using the R Package “rfishbase” 5.0.3. Two independent ancestral reconstruction analyses were carried out for both traits, “Mitogenome” and “Habitat”, using a reference evolutionary tree for teleosts to unravel both evolutionary histories. The possible association between mitogenome and habitat was then assessed using a suite of phylogenetic comparative methods, including Pagel’s correlation test (corHMM) to evaluate whether both traits evolved in a correlated fashion, branch-level co-transition analysis to identify lineages where structural changes and habitat shifts co-occurred, and node-by-node comparisons of ancestral state probabilities across the phylogeny. Preliminary results suggest a correlation between some deep-sea environments and a modified mitogenome structure, with structural deviations tending to cluster in lineages inhabiting greater depths. These exploratory findings raise the possibility that changes in mitogenome architecture may be linked to adaptations in energetic metabolism required for life in extreme low-energy environments. Further analyses are underway to clarify the functional significance of these genomic changes and their relationship to ecological and metabolic pressures in teleost evolution. 2026-06-18 Proceedings, Vol. 146, Pages 74: Widespread Gene Reorganizations in Teleost Mitochondria Are Driven by Ecological Transitions

Proceedings doi: 10.3390/proceedings2026146074

Authors: David Barros-García André Gomes-dos-Santos André M. Machado Francisco Baldó

The vertebrate mitochondrial genome (mitogenome) is a small, circular DNA molecule typically ~16–17 kb in length, encoding 37 genes that are essential for the electron transport chain, the mechanism that drives mostly all the ATP synthesis in cells. Owing to its central role in energy metabolism, its structure is highly conserved across vertebrate lineages in both the number and relative position of each gene in the genome. Nevertheless, different variations have been found in several teleost lineages, including antarctic fishes (Nototheniidae), gadiforms, hatchetfishes (Sternoptychidae), and Batrachoidiformes. The explanation for these phenomena remains unknown yet may reflect shifts in functional constraints and can provide insights into lineage-specific and/or coevolutionary processes. This raises the possibility that mitogenome structure is related to habitat selection, potentially reflecting environmental influences on energetic regulation. To further test this hypothesis, we studied more than 400 teleost species across all major teleost lineages. The mitogenome sequences were downloaded from NCBI and annotated using two independent algorithms (MITOZ and MITOS) and then compared with a reference (Danio rerio) to find any deviation from the standard structure. Similarly, ecological data was downloaded from FishBase using the R Package “rfishbase” 5.0.3. Two independent ancestral reconstruction analyses were carried out for both traits, “Mitogenome” and “Habitat”, using a reference evolutionary tree for teleosts to unravel both evolutionary histories. The possible association between mitogenome and habitat was then assessed using a suite of phylogenetic comparative methods, including Pagel’s correlation test (corHMM) to evaluate whether both traits evolved in a correlated fashion, branch-level co-transition analysis to identify lineages where structural changes and habitat shifts co-occurred, and node-by-node comparisons of ancestral state probabilities across the phylogeny. Preliminary results suggest a correlation between some deep-sea environments and a modified mitogenome structure, with structural deviations tending to cluster in lineages inhabiting greater depths. These exploratory findings raise the possibility that changes in mitogenome architecture may be linked to adaptations in energetic metabolism required for life in extreme low-energy environments. Further analyses are underway to clarify the functional significance of these genomic changes and their relationship to ecological and metabolic pressures in teleost evolution.

]]> Widespread Gene Reorganizations in Teleost Mitochondria Are Driven by Ecological Transitions David Barros-García André Gomes-dos-Santos André M. Machado Francisco Baldó doi: 10.3390/proceedings2026146074 Proceedings 2026-06-18 Proceedings 2026-06-18 146 1 Abstract 74 10.3390/proceedings2026146074 https://www.mdpi.com/2504-3900/146/1/74 Proceedings, Vol. 146, Pages 73: Rethinking Species Distribution Modelling for Freshwater Fish Under Environmental Changes https://www.mdpi.com/2504-3900/146/1/73 Introduction: Species Distribution Models (SDMs) are widely used to infer environmental drivers of freshwater fish distributions and to project biodiversity responses to climate and land-use change. However, freshwater ecosystems present specific conceptual and methodological challenges, including dendritic network structure, strong spatial autocorrelation, dispersal constraints, and scale mismatches between biological processes and environmental predictors that remain insufficiently addressed. At the same time, emerging data sources such as environmental DNA (eDNA) and high-resolution remote sensing offer new opportunities to improve data coverage and ecological realism in SDMs. Methodology: Focusing on Iberian systems as illustrative case studies, here, we synthesize the following recent advances and challenges in SDM applications to freshwater fishes: (i) the implications of using presence–absence versus abundance data; (ii) the integration of hydrological and connectivity metrics as predictors; (iii) approaches to explicitly account for spatial structure and biotic interactions; and (iv) the contribution of novel datasets, including eDNA and remote sensing. Furthermore, we examine the performance and transferability of correlative models under analogue and non-analogue climate conditions. Results: Our synthesis highlights the importance of incorporating network topology, seasonality, dispersal constraints, and novel data sources to improve ecological realism and predictive performance. The integration of emerging biodiversity and environmental data can substantially reduce data gaps and improve model calibration and validation, particularly in poorly sampled systems. Nonetheless, model transferability remains a challenge, particularly for endemic and range-restricted species. Advancing freshwater SDMs through the integration of hydrologically explicit frameworks and novel data sources will strengthen their capacity to support evidence-based management of freshwater fish assemblages facing accelerating environmental changes. 2026-06-18 Proceedings, Vol. 146, Pages 73: Rethinking Species Distribution Modelling for Freshwater Fish Under Environmental Changes

Proceedings doi: 10.3390/proceedings2026146073

Authors: Ana Filipa Filipe Janine da Silva Virgilio Hermoso

Introduction: Species Distribution Models (SDMs) are widely used to infer environmental drivers of freshwater fish distributions and to project biodiversity responses to climate and land-use change. However, freshwater ecosystems present specific conceptual and methodological challenges, including dendritic network structure, strong spatial autocorrelation, dispersal constraints, and scale mismatches between biological processes and environmental predictors that remain insufficiently addressed. At the same time, emerging data sources such as environmental DNA (eDNA) and high-resolution remote sensing offer new opportunities to improve data coverage and ecological realism in SDMs. Methodology: Focusing on Iberian systems as illustrative case studies, here, we synthesize the following recent advances and challenges in SDM applications to freshwater fishes: (i) the implications of using presence–absence versus abundance data; (ii) the integration of hydrological and connectivity metrics as predictors; (iii) approaches to explicitly account for spatial structure and biotic interactions; and (iv) the contribution of novel datasets, including eDNA and remote sensing. Furthermore, we examine the performance and transferability of correlative models under analogue and non-analogue climate conditions. Results: Our synthesis highlights the importance of incorporating network topology, seasonality, dispersal constraints, and novel data sources to improve ecological realism and predictive performance. The integration of emerging biodiversity and environmental data can substantially reduce data gaps and improve model calibration and validation, particularly in poorly sampled systems. Nonetheless, model transferability remains a challenge, particularly for endemic and range-restricted species. Advancing freshwater SDMs through the integration of hydrologically explicit frameworks and novel data sources will strengthen their capacity to support evidence-based management of freshwater fish assemblages facing accelerating environmental changes.

]]> Rethinking Species Distribution Modelling for Freshwater Fish Under Environmental Changes Ana Filipa Filipe Janine da Silva Virgilio Hermoso doi: 10.3390/proceedings2026146073 Proceedings 2026-06-18 Proceedings 2026-06-18 146 1 Abstract 73 10.3390/proceedings2026146073 https://www.mdpi.com/2504-3900/146/1/73 Proceedings, Vol. 146, Pages 64: AQUArestore: Advancing Dynamic Riverine Ecosystem Restoration Through Science–Community Co-Development https://www.mdpi.com/2504-3900/146/1/64 Introduction: AQUArestore is a three-year project focused on promoting adaptive ecological restoration strategies for river ecosystems in the vulnerable cross-border region of Portugal. The project responds to pressing environmental challenges across the territory, including severe habitat degradation, climate vulnerability, declining water security, and biodiversity loss, with particular concern for freshwater fish communities, making river restoration essential to preserve native species and freshwater ecosystem services. Objective: The project aims to develop a replicable framework for restoration of Mediterranean transboundary riverine habitats, supporting the objectives of the EU Nature Restoration Law (NRL, Regulation 2024/1991). The consortium AQUArestore will develop (1) robust restoration indicators, (2) implement living labs for restoration experimentation, and (3) establish capacity-building and training programs for technicians and citizens. Methodology: The project kick-off meeting was used to operationalize project tasks, detail the implementation calendar and milestones, and clarify responsibilities of each project member and partner institutions within the different work tasks. The meeting gathered consortium members from the coordinating institution CEF-ISA (researchers at the Instituto Superior de Agronomia) and partners WWF Portugal (an environmental NGO) and Mushmore Cooperative, each one contributing according to their respective expertise and institutional objectives. Results: The AQUArestore project kick-off meeting took place in January 2026 at ISA, Lisbon, and included a presentation of the NRL and a detailed discussion of project task development. In detail, the activities will begin with the compilation of information on previously restored sites (Task 1). This will support the development and validation of environmental and biodiversity indicators of restoration outcomes, including those linked to freshwater fish assemblages and riparian vegetation (Task 2). The project will then establish two living labs as platforms to test nature-based solutions in collaboration with stakeholders and local communities (Task 3). In parallel, AQUArestore will strengthen technical capacity through training for practitioners and public authorities (Task 4). Finally, dissemination will be supported through citizen science, communication activities, and stakeholder engagement, fostering a broader impact (Task 5). Together, these tasks provide an integrated, science-based, and participatory framework aiming to support adaptive river restoration under climate and environmental changes. Conclusions: By integrating ecological restoration, biodiversity and environmental monitoring, and stakeholder engagement, AQUArestore is expected to contribute to the recovery of Mediterranean freshwater ecosystems and improve habitat quality and connectivity for native fish communities, enhancing resilience to climate change and other anthropogenic pressures. 2026-06-18 Proceedings, Vol. 146, Pages 64: AQUArestore: Advancing Dynamic Riverine Ecosystem Restoration Through Science–Community Co-Development

Proceedings doi: 10.3390/proceedings2026146064

Authors: Ana Filipa Filipe Maria João Costa Arthur Cupertino Maria Teresa Ferreira Daniel Mameri Patricia María Rodríguez-González José M. Santos Catarina Grilo José Pedro Ramião João Oliveira

Introduction: AQUArestore is a three-year project focused on promoting adaptive ecological restoration strategies for river ecosystems in the vulnerable cross-border region of Portugal. The project responds to pressing environmental challenges across the territory, including severe habitat degradation, climate vulnerability, declining water security, and biodiversity loss, with particular concern for freshwater fish communities, making river restoration essential to preserve native species and freshwater ecosystem services. Objective: The project aims to develop a replicable framework for restoration of Mediterranean transboundary riverine habitats, supporting the objectives of the EU Nature Restoration Law (NRL, Regulation 2024/1991). The consortium AQUArestore will develop (1) robust restoration indicators, (2) implement living labs for restoration experimentation, and (3) establish capacity-building and training programs for technicians and citizens. Methodology: The project kick-off meeting was used to operationalize project tasks, detail the implementation calendar and milestones, and clarify responsibilities of each project member and partner institutions within the different work tasks. The meeting gathered consortium members from the coordinating institution CEF-ISA (researchers at the Instituto Superior de Agronomia) and partners WWF Portugal (an environmental NGO) and Mushmore Cooperative, each one contributing according to their respective expertise and institutional objectives. Results: The AQUArestore project kick-off meeting took place in January 2026 at ISA, Lisbon, and included a presentation of the NRL and a detailed discussion of project task development. In detail, the activities will begin with the compilation of information on previously restored sites (Task 1). This will support the development and validation of environmental and biodiversity indicators of restoration outcomes, including those linked to freshwater fish assemblages and riparian vegetation (Task 2). The project will then establish two living labs as platforms to test nature-based solutions in collaboration with stakeholders and local communities (Task 3). In parallel, AQUArestore will strengthen technical capacity through training for practitioners and public authorities (Task 4). Finally, dissemination will be supported through citizen science, communication activities, and stakeholder engagement, fostering a broader impact (Task 5). Together, these tasks provide an integrated, science-based, and participatory framework aiming to support adaptive river restoration under climate and environmental changes. Conclusions: By integrating ecological restoration, biodiversity and environmental monitoring, and stakeholder engagement, AQUArestore is expected to contribute to the recovery of Mediterranean freshwater ecosystems and improve habitat quality and connectivity for native fish communities, enhancing resilience to climate change and other anthropogenic pressures.

]]> AQUArestore: Advancing Dynamic Riverine Ecosystem Restoration Through Science–Community Co-Development Ana Filipa Filipe Maria João Costa Arthur Cupertino Maria Teresa Ferreira Daniel Mameri Patricia María Rodríguez-González José M. Santos Catarina Grilo José Pedro Ramião João Oliveira doi: 10.3390/proceedings2026146064 Proceedings 2026-06-18 Proceedings 2026-06-18 146 1 Abstract 64 10.3390/proceedings2026146064 https://www.mdpi.com/2504-3900/146/1/64 Proceedings, Vol. 146, Pages 67: Hepatic PCNA and Caspase-3 Dynamics in Brown Trout Across Reproductive Stages https://www.mdpi.com/2504-3900/146/1/67 Introduction: The liver is a central metabolic regulator responsible not only for energy storage and allocation but also for the production of essential egg proteins. In seasonal breeders such as brown trout, physiological demands increase during spawning, requiring strong metabolic and structural adaptations. Understanding the cellular mechanisms underlying these fluctuations is essential for clarifying how liver function is regulated during reproduction. Objective: The aim of this study was to investigate cellular mechanisms underlying liver remodelling during the reproductive cycle of male and female brown trout by examining proliferation and apoptosis markers at both molecular and protein levels during four distinct reproductive stages: spawning capable, regressing, regenerating, and developing. Methodology: Adult male and female brown trout were sampled at four reproductive stages. Liver gene expression of proliferating cell nuclear antigen (PCNA) and caspase-3 was analysed using qRT-PCR. At the protein level, liver tissue sections were immunolabelled with antibodies against PCNA and caspase-3. Caspase-3 staining intensity was quantified using ImageJ, while PCNA expression was assessed by counting positively stained nuclei. Results: PCNA gene expression was upregulated during the regressing stage, while PCNA-positive nuclei immunolabelling increased during the regressing and spawning-capable stages. Caspase-3 transcript levels showed no significant differences among stages, whereas immunostaining increased during regenerating and developing stages. Conclusions: Overall, the liver undergoes dynamic cellular remodelling throughout the reproductive cycle, balancing proliferation and apoptosis to meet reproductive demands. The results suggest that proliferation is more evident during regenerating stages, while apoptotic activity may be regulated post-transcriptionally. PCNA also indicates that proliferative activity may persist during spawning capable stages, reflecting ongoing mitotic readiness despite reduced overall proliferation. 2026-06-18 Proceedings, Vol. 146, Pages 67: Hepatic PCNA and Caspase-3 Dynamics in Brown Trout Across Reproductive Stages

Proceedings doi: 10.3390/proceedings2026146067

Authors: Amândio de Barros Diana Santos Tiago Lourenço Célia Lopes Tânia Vieira Madureira Eduardo Rocha

Introduction: The liver is a central metabolic regulator responsible not only for energy storage and allocation but also for the production of essential egg proteins. In seasonal breeders such as brown trout, physiological demands increase during spawning, requiring strong metabolic and structural adaptations. Understanding the cellular mechanisms underlying these fluctuations is essential for clarifying how liver function is regulated during reproduction. Objective: The aim of this study was to investigate cellular mechanisms underlying liver remodelling during the reproductive cycle of male and female brown trout by examining proliferation and apoptosis markers at both molecular and protein levels during four distinct reproductive stages: spawning capable, regressing, regenerating, and developing. Methodology: Adult male and female brown trout were sampled at four reproductive stages. Liver gene expression of proliferating cell nuclear antigen (PCNA) and caspase-3 was analysed using qRT-PCR. At the protein level, liver tissue sections were immunolabelled with antibodies against PCNA and caspase-3. Caspase-3 staining intensity was quantified using ImageJ, while PCNA expression was assessed by counting positively stained nuclei. Results: PCNA gene expression was upregulated during the regressing stage, while PCNA-positive nuclei immunolabelling increased during the regressing and spawning-capable stages. Caspase-3 transcript levels showed no significant differences among stages, whereas immunostaining increased during regenerating and developing stages. Conclusions: Overall, the liver undergoes dynamic cellular remodelling throughout the reproductive cycle, balancing proliferation and apoptosis to meet reproductive demands. The results suggest that proliferation is more evident during regenerating stages, while apoptotic activity may be regulated post-transcriptionally. PCNA also indicates that proliferative activity may persist during spawning capable stages, reflecting ongoing mitotic readiness despite reduced overall proliferation.

]]> Hepatic PCNA and Caspase-3 Dynamics in Brown Trout Across Reproductive Stages Amândio de Barros Diana Santos Tiago Lourenço Célia Lopes Tânia Vieira Madureira Eduardo Rocha doi: 10.3390/proceedings2026146067 Proceedings 2026-06-18 Proceedings 2026-06-18 146 1 Abstract 67 10.3390/proceedings2026146067 https://www.mdpi.com/2504-3900/146/1/67 Proceedings, Vol. 146, Pages 63: Thermal Modulation of Cytochrome P450 1A Immunostaining in Single and Mixture PAH-Exposed Brown Trout Hepatocytes https://www.mdpi.com/2504-3900/146/1/63 Introduction: Temperature is a key environmental factor influencing the physiological and biochemical processes of aquatic organisms, including xenobiotic metabolism. Understanding how temperature modulates the toxicological effects of pollutants such as polycyclic aromatic hydrocarbons (PAHs) is crucial in the context of climate change. Among these compounds, benzo[a]pyrene (BaP) and benzo[a]anthracene (BaA) are priority pollutants in aquatic environments, resulting from incomplete combustion. Their relevance is attributed to persistence and metabolic bioactivation potential. Fish primary hepatocyte cultures represent a relevant in vitro model for studying combined effects of thermal stress and chemical exposures, while supporting the 3Rs principles (Replacement, Reduction, and Refinement). Objective: This study aims to assess temperature-dependent effects of BaP and BaA, and their mixtures in brown trout hepatocytes using cytochrome P450 1A (CYP1A) immunohistochemistry as an indicator of xenobiotic metabolism. Methodology: Primary hepatocytes were isolated using a two-step collagenase perfusion method and cultured in 24-well plates at 18 °C and 22 °C. Cells were exposed for 72 h to supplemented L-15 medium (control) or to 0.1% dimethyl sulfoxide in supplemented L-15 medium (solvent control), as well as to single exposures of 1 and 10 µM of BaP and BaA and to equimolar mixtures of both compounds (1 and 10 µM). Viability was assessed using the lactate dehydrogenase (LDH) assay. CYP1A immunostaining was quantified based on cytoplasmic staining intensity relative to background area. Results: No significant effects on cell viability were observed under any condition. Temperature significantly reduced CYP1A expression in single exposures at 22 °C compared to 18 °C. BaP induced a significant dose-dependent increase, while BaA differed from controls only at 10 µM. In mixtures, only treatment- and dose-dependent effects were observed, with no temperature influence detected. Conclusions: Overall, the data highlight temperature as a key modulator of biochemical responses to PAHs, with single and mixed exposures eliciting distinct effects and suggesting potential synergism in mixtures. 2026-06-18 Proceedings, Vol. 146, Pages 63: Thermal Modulation of Cytochrome P450 1A Immunostaining in Single and Mixture PAH-Exposed Brown Trout Hepatocytes

Proceedings doi: 10.3390/proceedings2026146063

Authors: Rodrigo Alves Célia Lopes Rosária Seabra Sofia Esquível Maria J. Rocha Eduardo Rocha Tânia Vieira Madureira

Introduction: Temperature is a key environmental factor influencing the physiological and biochemical processes of aquatic organisms, including xenobiotic metabolism. Understanding how temperature modulates the toxicological effects of pollutants such as polycyclic aromatic hydrocarbons (PAHs) is crucial in the context of climate change. Among these compounds, benzo[a]pyrene (BaP) and benzo[a]anthracene (BaA) are priority pollutants in aquatic environments, resulting from incomplete combustion. Their relevance is attributed to persistence and metabolic bioactivation potential. Fish primary hepatocyte cultures represent a relevant in vitro model for studying combined effects of thermal stress and chemical exposures, while supporting the 3Rs principles (Replacement, Reduction, and Refinement). Objective: This study aims to assess temperature-dependent effects of BaP and BaA, and their mixtures in brown trout hepatocytes using cytochrome P450 1A (CYP1A) immunohistochemistry as an indicator of xenobiotic metabolism. Methodology: Primary hepatocytes were isolated using a two-step collagenase perfusion method and cultured in 24-well plates at 18 °C and 22 °C. Cells were exposed for 72 h to supplemented L-15 medium (control) or to 0.1% dimethyl sulfoxide in supplemented L-15 medium (solvent control), as well as to single exposures of 1 and 10 µM of BaP and BaA and to equimolar mixtures of both compounds (1 and 10 µM). Viability was assessed using the lactate dehydrogenase (LDH) assay. CYP1A immunostaining was quantified based on cytoplasmic staining intensity relative to background area. Results: No significant effects on cell viability were observed under any condition. Temperature significantly reduced CYP1A expression in single exposures at 22 °C compared to 18 °C. BaP induced a significant dose-dependent increase, while BaA differed from controls only at 10 µM. In mixtures, only treatment- and dose-dependent effects were observed, with no temperature influence detected. Conclusions: Overall, the data highlight temperature as a key modulator of biochemical responses to PAHs, with single and mixed exposures eliciting distinct effects and suggesting potential synergism in mixtures.

]]> Thermal Modulation of Cytochrome P450 1A Immunostaining in Single and Mixture PAH-Exposed Brown Trout Hepatocytes Rodrigo Alves Célia Lopes Rosária Seabra Sofia Esquível Maria J. Rocha Eduardo Rocha Tânia Vieira Madureira doi: 10.3390/proceedings2026146063 Proceedings 2026-06-18 Proceedings 2026-06-18 146 1 Abstract 63 10.3390/proceedings2026146063 https://www.mdpi.com/2504-3900/146/1/63 Proceedings, Vol. 146, Pages 71: Exploration of Enantioselective Effects of MDPV on Zebrafish Embryogenesis https://www.mdpi.com/2504-3900/146/1/71 Introduction: Synthetic cathinones (SC) are an emerging class of neuroactive contaminants increasingly detected in aquatic systems due to their widespread recreational use. Their continuous release at ng–µg L−1 levels is particularly concerning, as these compounds are specifically designed to alter neural function, raising the likelihood of subtle yet ecologically relevant effects in non-target organisms. Among them, 3,4-methylenedioxypyrovalerone (MDPV) is one of the most-reported SC in wastewater and surface waters. Nevertheless, its chiral nature has been largely overlooked in ecotoxicological studies, despite growing evidence that enantiomers can differ markedly in biological activity, potentially leading to underestimated environmental risks. Objective: The ecotoxicological impact of racemic MDPV ((R,S)-MDPV) and its separate enantiomers ((R)-MDPV and (S)-MDPV) were examined using zebrafish (Danio rerio) as a model, focusing on survival and embryonic development. Methodology: Zebrafish embryos, at approximately 3-hours post-fertilization (hpf), were exposed over 96 h to environmentally relevant concentrations of MDPV forms (0.18−2.8 μg L−1). Each treatment and control group included 50 animals distributed across 5 replicates. Mortality was assessed at multiple developmental stages (7, 24, 48, 72, and 96 h), along with cumulative mortality. Developmental endpoints included spontaneous movements (24 h), heartbeat (48 h), and hatching rate (48 and 72 h), quantified using stereomicroscopy and video analysis. Results: MDPV showed concentration and enantioselective effects, with (S)-MDPV being the most toxic. Behavioral and cardiac responses varied across forms, while hatching depended on concentration and time without a clear enantioselective pattern. Conclusions: MDPV disrupts early zebrafish development, impairing survival and embryonic development in a concentration-dependent and enantioselective manner, with (S)-MDPV demonstrating greater toxicity. These findings emphasize the importance of considering chirality in the environmental risk assessment of psychoactive contaminants such as SC, as enantiomer-specific effects may influence organism fitness, survival, and broader ecological outcomes. 2026-06-18 Proceedings, Vol. 146, Pages 71: Exploration of Enantioselective Effects of MDPV on Zebrafish Embryogenesis

Proceedings doi: 10.3390/proceedings2026146071

Authors: Ariana Pérez-Pereira Ondina Ribeiro Luís Félix Maria Tiritan Cláudia Ribeiro João Carrola

Introduction: Synthetic cathinones (SC) are an emerging class of neuroactive contaminants increasingly detected in aquatic systems due to their widespread recreational use. Their continuous release at ng–µg L−1 levels is particularly concerning, as these compounds are specifically designed to alter neural function, raising the likelihood of subtle yet ecologically relevant effects in non-target organisms. Among them, 3,4-methylenedioxypyrovalerone (MDPV) is one of the most-reported SC in wastewater and surface waters. Nevertheless, its chiral nature has been largely overlooked in ecotoxicological studies, despite growing evidence that enantiomers can differ markedly in biological activity, potentially leading to underestimated environmental risks. Objective: The ecotoxicological impact of racemic MDPV ((R,S)-MDPV) and its separate enantiomers ((R)-MDPV and (S)-MDPV) were examined using zebrafish (Danio rerio) as a model, focusing on survival and embryonic development. Methodology: Zebrafish embryos, at approximately 3-hours post-fertilization (hpf), were exposed over 96 h to environmentally relevant concentrations of MDPV forms (0.18−2.8 μg L−1). Each treatment and control group included 50 animals distributed across 5 replicates. Mortality was assessed at multiple developmental stages (7, 24, 48, 72, and 96 h), along with cumulative mortality. Developmental endpoints included spontaneous movements (24 h), heartbeat (48 h), and hatching rate (48 and 72 h), quantified using stereomicroscopy and video analysis. Results: MDPV showed concentration and enantioselective effects, with (S)-MDPV being the most toxic. Behavioral and cardiac responses varied across forms, while hatching depended on concentration and time without a clear enantioselective pattern. Conclusions: MDPV disrupts early zebrafish development, impairing survival and embryonic development in a concentration-dependent and enantioselective manner, with (S)-MDPV demonstrating greater toxicity. These findings emphasize the importance of considering chirality in the environmental risk assessment of psychoactive contaminants such as SC, as enantiomer-specific effects may influence organism fitness, survival, and broader ecological outcomes.

]]> Exploration of Enantioselective Effects of MDPV on Zebrafish Embryogenesis Ariana Pérez-Pereira Ondina Ribeiro Luís Félix Maria Tiritan Cláudia Ribeiro João Carrola doi: 10.3390/proceedings2026146071 Proceedings 2026-06-18 Proceedings 2026-06-18 146 1 Abstract 71 10.3390/proceedings2026146071 https://www.mdpi.com/2504-3900/146/1/71