Pathogens doi: 10.3390/pathogens15070766

Authors: Francesco Broccolo Alessandro Sannino Mauro Pollini Federica Paladini Thierry Mourer Francesca Di Nunzio

Theories on the early steps of the HIV-1 life cycle have been radically revised over the past five years. The long-held assumption that the capsid fully disassembles in the cytoplasm has given way to a more nuanced view: Cytoplasmic disassembly does occur and, in several myeloid systems, is increasingly linked to cytosolic cDNA sensing and to abortive infection. However, a substantial fraction of intact or nearly intact capsid cores instead traverse the nuclear pore complex (NPC) and, upon interacting with the host factor CPSF6, induce liquid–liquid phase separation. This leads to the formation of biomolecular condensates, termed HIV-1 membraneless organelles (HIV-1-MLOs), which subsequently merge with nuclear speckles (NSs). In this Perspective we read these condensates along five interlocking axes. First, the virus drives the host phase separation of cleavage and polyadenylation specificity factor 6 (CPSF6), which quickly fuses with another MLO: the NS composed of the speckle scaffold factors, SON and SRRM2. Second, the resulting condensate behaves as a catalytic site that concentrates the reverse-transcription machinery and thereby promotes integration of the viral DNA into speckle-associated chromatin (SPADs). Third, the same compartment is the final layer of a stratified programme of innate immune evasion, shielding nascent double-stranded DNA from cGAS–STING after cytoplasmic restriction factors and sensors have been outmanoeuvred. Fourth, although demonstrated only in vitro, stable HIV-1-MLOs can maintain the viral RNA genome in the presence of a reverse-transcription inhibitor. Upon removal of the inhibitor, reverse transcription resumes, mirroring, to some extent, the situation in individuals undergoing interruption of antiretroviral therapy and suggesting that these structures may act as a pre-integration reservoir. Fifth, and still largely unexplored, the sanctuary has a pharmacological dimension: anatomical lymphoid compartments, and possibly the condensate itself through selective small-molecule partitioning, may limit antiretroviral drug access. We situate HIV-1-MLOs within the convergent condensate strategies of SARS-CoV-2 and other viruses, and we discuss the clinical, diagnostic, therapeutic, and vaccine implications, including capsid inhibitors as “block-and-expose” tools.

Pathogens doi: 10.3390/pathogens15070765

Authors: Hossein Valishirin Sai Deepika Reddy Yaram Negar Farhang Doost Soumya K. Srivastava Shira L. Broschat

Tick-borne bacterial infections such as those caused by Anaplasma phagocytophilum are difficult to diagnose, particularly at early stages, because they rely on specialized laboratory tests. Dielectric-based measurements provide a label-free way to probe cellular state and may offer useful information. In this study, we explored whether A. phagocytophilum-infected and uninfected HL-60 human promyelocytic leukemia cells can be distinguished using dielectric measurements collected under controlled in vitro conditions. Measurements were performed at two medium conductivities (100 and 300 µS/cm), and three dielectric properties were examined: cytoplasmic conductivity, specific membrane conductance, and specific membrane capacitance. We used exploratory analysis, statistical tests, and interpretable linear classifiers, with performance evaluated using leave-one-out cross-validation. Membrane-related properties, especially specific membrane conductance, showed the clearest separation between infection states and were consistently the most informative across models. Both classification performance and effect sizes were stronger at the higher medium conductivity condition. These results are encouraging, but because the sample size was small and consisted only of technical replicates, they should be interpreted as preliminary measurement-level evidence. This study is intended as a pilot and supports further work with larger datasets, additional cell types, and broader experimental conditions.

Pathogens doi: 10.3390/pathogens15070764

Authors: Abel S. Biguezoton Chaka Traore Haladou Gagara Der Dabire Zakaria Bengaly Mahaman Maaouia Abdou Moussa Raïssa Bakinahe Ntamukunzi Kader Issoufou Maïmouna Ousmane Claude Saegerman Marcella Mori

As part of the PRISMA project, which promotes research and innovation for productive, resilient, and healthy agro-pastoral systems in West Africa, a cross-sectional survey was conducted to assess the apparent seroprevalence and investigate epidemiological factors associated with bovine brucellosis in dairy herds. The survey was carried out in Mali (Bamako, Koulikoro, Mopti, and Sikasso regions) and Niger (Tahoua, Dosso, and Tillabéry regions). A total of 1230 animals from 82 herds were tested, including 645 animals from 43 herds in Mali and 585 animals from 39 herds in Niger. At the herd level, apparent seroprevalence was significantly higher in Mali than in Niger (37.21% vs. 15.38%). Within each country, animal-level seroprevalence varied across region, with the highest levels observed in Bamako (11.11%) and Mopti (7.62%) in Mali, and Dosso (6.67%) in Niger. High within-herd prevalence (>20%) was identified in only one herd per country. The higher overall herd-level seroprevalence in Mali was primarily driven by a greater number of herds with low (≤10%) within-herd seroprevalence. Univariate risk factor analysis at the animal level identified a borderline trend toward higher odds of seropositivity in crossbred cattle than other breeds (OR = 1.95, 95% CI: 1.00–3.81, p = 0.0503). In addition, a classification tree analysis highlighted seven exploratory signals of the positive herd status. These included four animal/management-level factors—herd region, environmental disposal of aborted placentas, the number of lactating cows (as a proxy for animal density), and reported herd abortions—as well as three human-level signals within the household: a history of miscarriage, experiencing heavy night sweats, and suffering from undulant fever. In conclusion, this survey provides new insights into bovine brucellosis and its epidemiology in dairy herds in Mali and Niger, confirming established high-prevalence areas such as Bamako and identifying new zones of concern, such as Mopti and Dosso. Our findings underscore that bovine brucellosis remains an important public health and economic threat in both countries. Given that the identified positive regions are hubs for milk production and consumption, these data may support health authorities in implementing targeted surveillance, control, and prevention strategies under a One Health approach.

Pathogens doi: 10.3390/pathogens15070763

Authors: Matteo Fortunato Anne Tabard-Fougère Giacomo De Marco Oscar Vazquez Christina Steiger Elio Paris Ardian Ramadani Andreas Tsoupras Romain Dayer Dimitri Ceroni

Kingella kingae is now recognised as the leading cause of osteoarticular infections (OAIs) in young children. Because these infections typically produce mild symptoms and a weak inflammatory response, early diagnosis remains challenging. Complete blood count-derived immune-inflammatory biomarkers (IIBs) have recently gained interest as accessible, low-cost indicators of systemic inflammation, with potential diagnostic value in several fields, including oncology, rheumatology, cardiology, and infectious diseases. This study therefore aimed to assess whether IIBs could support the screening of K. kingae OAIs. We retrospectively reviewed the medical records of 209 children admitted to our hospital between 2007 and 2025 with confirmed or highly suspected K. kingae OAIs. Complete blood counts were analysed for each patient, including leukocyte subtypes such as neutrophils, lymphocytes, monocytes, eosinophils, and basophils. We then calculated biomarkers (NLR, MLR, PLR, SII, SIRI, and PIV) and interpreted them using age-adjusted reference values. As expected, most children were younger than 48 months, and septic arthritis was the predominant clinical presentation. Classical acute-phase reactants (WBC, CRP, and ESR) were frequently normal or only mildly elevated. Likewise, most IIBs remained within or near reference ranges; the platelet-to-lymphocyte ratio was the most commonly abnormal marker, exceeding the threshold in only 59.3% of patients. These results indicate that, like conventional inflammatory markers, CBC-derived IIBs have limited standalone screening utility for K. kingae OAIs. Further studies are needed to determine whether systemic IIBs can help distinguish OAIs caused by K. kingae from those caused by pyogenic bacteria.

Pathogens doi: 10.3390/pathogens15070762

Authors: Branislav Milovanović Nikola Marković Elizabeta Ristanović Nikoleta Đorđevski Sonja Atanasievska Kujović Sulin Bulatović Vasko Žugić Maša Petrović Milovan Bojić

Background: Bartonella henselae infection has been associated with a broad spectrum of neurological and autonomic manifestations, although its impact on autonomic nervous system function remains insufficiently characterized, and the aim of this study was to evaluate autonomic function in patients with polymorphic symptoms and Bartonella henselae IgM seroreactivity. Methods: In this cross-sectional study, 75 patients were compared with 75 age- and sex-matched healthy controls, and all participants underwent cardiovascular autonomic reflex testing, short-term (5 min) and long-term (24 h) heart rate variability analysis, and 24 h ambulatory blood pressure monitoring, while head-up tilt testing was performed in the Bartonella group. Results: Abnormal autonomic reflex tests were significantly more frequent in the Bartonella IgM-seroreactive group, particularly those reflecting parasympathetic function, while heart rate variability analysis demonstrated reduced high-frequency components and lower long-term variability indices, and head-up tilt testing revealed heterogeneous hemodynamic responses including orthostatic hypotension and pronounced blood pressure variability, with ambulatory monitoring additionally showing higher nighttime blood pressure values and reduced nocturnal dipping. Conclusions: These findings indicate a consistent pattern of autonomic imbalance characterized predominantly by parasympathetic impairment and altered blood pressure regulation in patients with Bartonella henselae IgM seroreactivity, although further studies are required to clarify underlying mechanisms and clinical implications.

Pathogens doi: 10.3390/pathogens15070761

Authors: Hathem Khelil Rosanna Palumbo Giovanni N. Roviello

Artificial intelligence (AI) has rapidly emerged as a transformative tool in virology, offering new opportunities for the detection, classification, and surveillance of viral pathogens. Recent advances in machine learning, deep neural networks, and multimodal data analysis now enable the identification of viral signatures from genomic sequences, medical images, environmental samples, and social-media-derived epidemiological signals. This review provides a comprehensive overview of state-of-the-art AI methodologies applied to viral pathogen research, with a particular focus on image-based diagnostics, automated quality assessment of virology-related digital content, and predictive modelling for outbreak monitoring. We discuss how convolutional and transformer-based architectures are being used to classify infected tissues, detect viral particles, and support laboratory workflows. Furthermore, we highlight the emerging role of AI in evaluating the reliability of user-generated images and short videos related to infectious diseases, an area increasingly relevant in the age of misinformation. Challenges such as dataset bias, limited annotated virological images, ethical concerns, and the need for standardized quality-assessment pipelines are critically examined. Finally, we outline future research directions, including hybrid AI–biological models, AI-supported viral surveillance in healthcare environments, and the integration of explainable AI to enhance clinical trust.

Pathogens doi: 10.3390/pathogens15070760

Authors: Miriana Quaranta Alessandra Ciccozzi Francesco Branda Leonardo Sernicola Massimo Ciccozzi Stefano Pascarella Alessandra Borsetti Fabio Scarpa

In November 2024, a highly mutated descendant of the Omicron BA.3 subvariant, designated BA.3.2, emerged in South Africa carrying 39 spike mutations, two large N-terminal domain (NTD) deletions and a novel four-amino acid insertion. A key feature of BA.3.2 is extensive NTD remodeling, including a major deletion spanning residues 135–148 affecting the β-hairpin region and contributing to the loss of most of the N1 loop. This study compares the evolutionary dynamics and structural features of BA.3.2 with BA.3. Phylodynamic analyses show that BA.3 underwent early demographic stability followed by a decline in genetic diversity, consistent with limited circulation, whereas BA.3.2 displays recent emergence and a progressive reduction in effective population size without rapid expansion. Selection analyses indicate BA.3 evolution is mainly driven by changes in the receptor-binding domain, while BA.3.2 shows dispersed signals across spike regions, including codon 1162. Structural and molecular dynamic analyses reveal increased flexibility and a broader conformational landscape in the BA.3.2 NTD, driven by the deletion and resulting loss of stabilizing interactions. Overall, BA.3.2 follows a distinct evolutionary trajectory characterized by antigenic remodeling of the spike NTD, underlining the need for continued surveillance of emerging SARS-CoV-2 descendant lineages.

Pathogens doi: 10.3390/pathogens15070759

Authors: Pavlo Petakh Iryna Halabitska Oleh Lushchak Oleksandr Kamyshnyi

Background: Ukraine has substantial regional differences in climate, landscape, forest cover, and river networks, which may influence leptospirosis transmission. However, recent monthly oblast-level variation in leptospirosis incidence has not been systematically described. We used newly available surveillance data for 2023–2025 to assess seasonal and regional patterns of leptospirosis and their associations with weather, climatic zone, forest cover, and river network density. Methods: We analyzed surveillance data from 23 Ukrainian oblasts. Incidence was assessed by month, oblast, and climatic zone. Weather data were aggregated monthly; forest cover and river network density were included as predictors. Associations were assessed using correlation, cross-correlation, and Random Forest ML models. Results: Incidence showed a clear seasonal increase, reaching its highest levels in late summer and autumn, approximately one to three months after seasonal peaks in temperature and precipitation. The highest incidence was observed in Zakarpattia, Chernihiv, and Ternopil oblasts, while incidence by climatic zone was highest in the Carpathian group and lowest in the Steppe zone. Among weather variables, average temperature showed the clearest delayed association with leptospirosis incidence. River network density was the leading ecological predictor in the adjusted models. The positive unadjusted association between forest cover and incidence became negative after adjustment for river network density, suggesting that these variables captured overlapping ecological characteristics. In the Random Forest analysis, river network density was the top-ranked predictor, and the best-performing model achieved an AUC of 0.795. Conclusions: Leptospirosis incidence in Ukraine varied substantially by season and region. Delayed temperature effects, river network density, and forest cover were associated with regional risk. Monthly oblast-level surveillance with climatic and ecological data may help monitor leptospirosis risk, but war-related disruption to diagnosis and reporting should be considered.

Pathogens doi: 10.3390/pathogens15070758

Authors: Umar Saeed Rizwan Uppal Gohar Zaman Muhammad Rehan Uppal Zsolt Jenő Szepesváry Aftab Ahmad Khan Muhammad Usman Qamar Zuhaib Ali Zahra Zahid Piracha

Urinary tract infections are among the most common bacterial infections encountered in clinical practice, with Escherichia coli representing the dominant urinary pathogen. Increasing detection of multidrug-resistant and extended-spectrum beta-lactamase (ESBL)-producing uropathogens has narrowed empirical treatment options and renewed interest in fosfomycin. However, local long-term surveillance data on fosfomycin susceptibility remain limited in Pakistan. This study evaluated temporal changes in major urinary isolate categories and fosfomycin susceptibility patterns within a diagnostic laboratory network in Pakistan from 2013 to 2025. An exploratory molecular sub-analysis was also performed to assess selected resistance-associated transcript patterns in archived fosfomycin-susceptible and fosfomycin-resistant isolates. A retrospective laboratory-based, isolate-level analysis was conducted using anonymized urine culture records. The source database included 34,230 urine sample records, from which eligible culture-positive urinary isolates with required organism classification and fosfomycin susceptibility data were included in the final analytical dataset. Analyses were performed across predefined mutually exclusive study intervals. Organism categories included non-ESBL E. coli, ESBL-producing E. coli, laboratory-coded ESBL E. coli 24 variant, Klebsiella spp., and Enterococcus spp. The ESBL E. coli 24 variant was treated as a laboratory reporting category, not as a genomically confirmed clone or sequence type. Fosfomycin resistance was evaluated using interval-based comparisons and odds ratios. A selected subset of 24 archived isolates, including fosfomycin-susceptible and fosfomycin-resistant E. coli and Klebsiella pneumoniae, was analyzed by RT-qPCR for glpT, uhpT, murA, fosA, fosA3, and blaCTX-M transcript abundance. The final isolate-level analytical dataset included 17,978 eligible urinary isolates. Among urine records with available sex data, female-associated records represented the majority throughout the study period, but this finding reflects laboratory record distribution rather than patient-level UTI prevalence. E. coli remained the predominant urinary isolate category. Non-ESBL E. coli declined across study intervals, whereas ESBL-associated E. coli categories represented a larger proportion of isolates in later years. The laboratory-coded ESBL E. coli 24 variant increased in later intervals, although this finding requires cautious interpretation because confirmatory molecular typing was not performed. Fosfomycin resistance showed a non-linear temporal pattern: resistance decreased from the early to the middle interval and then increased markedly to 23.8% during 2021–2025, while susceptibility declined to 60.6% in the same interval. Compared with the middle interval, isolates from 2021–2025 had higher odds of fosfomycin resistance (OR = 3.64, 95% CI: 3.23–4.12; p < 0.001). In the exploratory molecular subset, resistant isolates showed lower transcript abundance of selected uptake-associated genes, particularly glpT and uhpT, and higher expression of selected fosfomycin- and ESBL-associated genes, including fosA, fosA3, and blaCTX-M. These findings represent transcriptional associations in selected isolates and do not establish definitive resistance mechanisms. Urinary isolates in this diagnostic-network dataset showed a temporal shift toward greater representation of laboratory-reported ESBL-associated E. coli categories and a marked increase in fosfomycin resistance during 2021–2025. The findings support continued local surveillance of urinary pathogens and periodic reassessment of fosfomycin susceptibility for antimicrobial-stewardship guidance. The molecular findings should be interpreted as exploratory transcriptional observations because they were based on a small selected isolate subset and were not supported by genomic, mutational, uptake, or functional validation.

Pathogens doi: 10.3390/pathogens15070757

Authors: Joaquín Saavedra Marcelo Gómez Pamela Muñoz Valentina Bernal Pedro Aburto Eduardo Raffo Brandon Aristizabal Manuel Moroni Gisella Paredes Andrea Monterroza Rodrigo Arancibia Anja Taubert Carlos Hermosilla

Background: The American mink (Neogale vison) is an invasive mustelid in Chile that serves as a natural reservoir host for several pathogens. Cardiorespiratory nematodes of the superfamily Metastrongyloidea, including Aelurostrongylus spp., have been reported in carnivores worldwide but remain poorly documented in South American wildlife and invasive species. This study reports the first detection of Aelurostrongylus spp. infection in American minks in Southern Chile. Methods: Between 2022 and 2024, 27 American mink carcasses were collected from the Los Ríos and Los Lagos regions, Southern Chile. Complete necropsies were performed, and cardiorespiratory organs were examined for presence of parasites. Nematode larvae identified in bronchoalveolar lavage were characterized both morphologically and molecularly through PCR amplification, confirming their classification within the superfamily Metastrongyloidea. Results: Metastrongyloid nematodes were detected in 33.3% (9/27) of examined mink. Histopathological examination of lung tissue in some specimens identified adult parasite specimens. Morphological examination revealed Aelurostrongylus spp. first-stage larvae (L1) with characteristic morphology and morphometry. Molecular analysis confirmed the presence of Aelurostrongylus spp., with sequences showing 99% identity to Aelurostrongylus abstrusus. Most of the infected individuals were adults collected from freshwater environments. Conclusions: This study provides the first evidence of patent Aelurostrongylus spp. infection in neozoan American minks in South America, highlighting the role of this invasive species as a potential reservoir for cardiorespiratory parasites and emphasizing the need for continued surveillance of parasitic infections originating from invasive carnivores.

Pathogens doi: 10.3390/pathogens15070756

Authors: Djana Rrupa Christelle Kaiser Emmanuel Melloul Emilie Uldry Nermin Halkic Guido Beldi Severin Gloor Anja Lachenmayer Gaëtan-Romain Joliat

Long-term outcomes after hepatectomy for alveolar echinococcosis (AE) in immunosuppressed patients remain poorly investigated. This study evaluated recurrence rates and recurrence-free/overall survivals (RFS/OS) after AE resection in immunocompromised and immunocompetent patients. Consecutive patients operated for liver AE in two university hospitals were retrospectively collected (2000–2021). Immunosuppressed patients were defined as patients who had a reduced ability to fight infection due to certain diseases or treatments. 195 patients had hepatectomy for liver AE. Preoperative albendazole was given in 111 cases (57%). Fifty-two patients (27%) were considered immunosuppressed. Patients in the immunosuppressed and immunocompetent cohorts had similar preoperative characteristics. Recurrences occurred in 10 patients (immunosuppressed group: 4, non-immunosuppressed group: 6) within a median follow-up of 58 months (95%CI 48–68). No significant differences in RFS and OS were found between the immunosuppressed and immunocompetent groups (212 vs. 206 months, p = 0.625 and 236 vs. 210 months, p = 0.282). Two-year recurrence rates were 0% in the immunosuppressed cohort and 1% (1/142) in the immunocompetent patients (p = 0.466). Absence of preoperative albendazole and lesion size were predictive of recurrence after hepatectomy. Immunosuppression was not found to be a risk factor for recurrence (HR 1.4, p = 0.626). In this bicentric study, immunocompromised patients did not have significantly different recurrence rates, RFS, and OS than immunocompetent patients.

Pathogens doi: 10.3390/pathogens15070755

Authors: Yaqi Xie Quanman Hu Shuaiyin Chen Baoqin Zhang

Objective: The purpose of this study is to study cytokine indicators for the identification of severe fever with thrombocytopenia syndrome (SFTS) severity. Methods: We searched the literature in PubMed, Embase, and Web of Science published before 7 April 2026. The main results are presented as forest plots. Subgroup analyses, sensitivity analyses, and publication bias were also performed. Results: A total of 22 articles were eventually included in our study. Our findings demonstrate that circulating concentrations of Interleukin-6 (IL-6) (SMD = 2.02, 95% CI: 1.53–2.51, I2 = 95.2%), Interleukin-10 (IL-10) (SMD = 1.18, 95% CI: 0.92–1.44, I2 = 71.3%), Interleukin-8 (IL-8) (SMD = 0.91, 95% CI: 0.61–1.20, I2 = 69%), Tumor necrosis factor-alpha (TNF-α) (SMD = 0.70, 95% CI: 0.43–0.96, I2 = 64.6%), Interferon-gamma (IFN-γ) (SMD = 1.32, 95% CI: 0.69–1.95, I2 = 89.1%), Interleukin-1 beta (IL-1β) (SMD = 1.78, 95% CI: 0.85–2.71, I2 = 94.8%), Monocyte chemoattractant protein-1 (MCP-1) (SMD = 1.15, 95% CI: 0.80–1.50, I2 = 46.1%), Interferon-alpha (IFN-α) (SMD = 1.53, 95% CI: 0.38–2.68, I2 = 89.6%), Granulocyte Colony-Stimulating Factor (G-CSF) (SMD = 1.79, 95% CI: 0.99–2.59, I2 = 68.1%) and Inducible protein 10 (IP-10) (SMD = 1.11, 95% CI: 0.60–1.62, I2 = 58.3%) are significantly elevated in patients with severe SFTS compared with those with mild disease, whereas Transforming Growth Factor-beta (TGF-β) (SMD = −0.51, 95% CI: −0.78–−0.24, I2 = 6.0%) and RANTES (SMD = −0.10, 95% CI: −0.40–0.20, I2 = 0.0%) levels are reduced in the severe group. Conclusions: By analyzing the cytokine indicators of SFTS patients, we have found some indicators that are representative of SFTS severity. Our findings provide a clinically actionable basis for early severity prediction and further useful evidence for clinicians to manage severe patients efficiently.

Pathogens doi: 10.3390/pathogens15070754

Authors: Yuhan Ding Chao Huang Shuo Ning Jingxian Liu Yiyue Ge Ying Chi

Protein phosphorylation is a central post-translational modification. In pathogenic fungi, it dynamically governs morphogenesis, stress adaptation, and antifungal drug resistance. Using high-resolution mass spectrometry-based phosphoproteomics, researchers have systematically mapped phosphorylation dynamics in WHO-priority pathogens—Candida albicans, Aspergillus fumigatus, Cryptococcus neoformans, and the multidrug-resistant Candidozyma auris (formerly Candida auris). These studies reveal that thousands of phosphorylation events are coordinately reprogrammed in response to antifungal drug exposure, host-derived oxidative stress, and temperature shifts. Integration of available datasets suggests a “conserved-core/divergent-output” organization. Shared kinase hubs like cAMP-PKA, HOG-MAPK and calcineurin are broadly conserved across species. Downstream substrate networks, however, have diverged, producing distinct virulence outputs in each pathogen. Notably, C. auris remains completely uncharacterized at the phosphoproteomic level. This review provides a comprehensive synthesis of the phosphoproteomic landscape across these pathogens, and discusses how phosphoproteomic data are guiding the rational prioritization of kinases and phosphatases as next-generation antifungal drug targets—with direct implications for clinical surveillance and public health.

Pathogens doi: 10.3390/pathogens15070753

Authors: Karolina M. Czajka Asma Bilgasem Yamamah A. Al-Jumaili Denver Kitching Graham Buchan Anu Nair Michael Reich Chibike Ijomah Gopi E. Saikrishna Chris Verschoor Stacey A. Santi Danielle Brabant-Kirwan Ravi Singh Vasu Appanna Deborah Saunders Sujeenthar Tharmalingam

Rapid and accurate identification of Candida species is critical for guiding antifungal therapy, especially with the emergence of intrinsically resistant pathogens. However, diagnostics using culture-based methods remain slow and labor-intensive, limiting timely treatment decisions. This systematic review evaluated the diagnostic performance and clinical applicability of nucleic acid amplification tests (NAATs) for Candida species identification using a PubMed search completed on 23 June 2025. A total of 888 records were screened, 333 full-text articles were assessed, and 158 studies were included based on criteria including comparison with standard diagnostic methods, diagnostic performance reporting, and involvement of clinical samples. PCR-based approaches were the most widely used, including conventional, nested, multiplex, real-time, and droplet digital PCR. Isothermal methods such as loop-mediated isothermal amplification (LAMP) and recombinase polymerase amplification (RPA) were increasingly represented. Common molecular targets included the ITS and 18S/28S rDNA regions. The risk of bias assessment was completed with the QUADAS-2 tool. Diagnostic performance varied depending on assay design, specimen type, gene target, and reference method. Excellent specificity and low limits of detection were achieved, especially with isothermal platforms offering the shortest turnaround times and greatest potential for point-of-care implementation. Multiplex assays were particularly advantageous for detecting mixed-species samples, while highly specific assays were optimal for distinguishing clinically important species such as Candidozyma auris, Nakaseomyces glabratus, and Pichia kudriavzevii. Overall, NAATs represent a promising diagnostic tool for Candida species identification, but broader clinical adoption will require improved standardization, validation across diverse patient populations, and clearer interpretation of fungal burden in the context of colonization versus infection.

Pathogens doi: 10.3390/pathogens15070752

Authors: Maowen Sun Cong Yuan Yu Zhang Xueliang Zhu Lei Shi Yueyue Duan Wenquan Mao Luyao Li Yanghe Liu Qi Wang

The emergence of swine acute diarrhea syndrome coronavirus (SADS-CoV), an alpha-coronavirus that causes fatal enteric disease in neonatal piglets with mortality rates up to 90%, demonstrates that bat-origin coronavirus has expanded its host range to pigs. Although SADS-CoV exhibits significant pandemic potential and capacity for cross-species transmission, the replication mechanisms of SADS-CoV remain largely unexplored. Identifying host factors responsible for SADS-CoV replication and elucidating its underlying mechanisms is essential for advancing fundamental knowledge of coronavirus biology and developing antiviral therapies. Here, we identified PABPC1 as a novel interactor of the SADS-CoV nucleocapsid (N) protein. Overexpression of PABPC1 restricted SADS-CoV infection, whereas knockdown of PABPC1 enhanced viral replication. Further study indicated PABPC1 as a host restriction factor of SADS-CoV in a manner dependent on its PABC domain. Mechanistically, PABPC1 enhances the ubiquitination of the N protein, therefore facilitating its recognition by the cargo receptor TOLLIP for selective autophagic degradation. This study systematically analyzes the interaction of host factors and the SADS-CoV N protein and identifies PABPC1 as a host restriction factor that limits viral replication via TOLLIP-mediated selective autophagy degradation of the N protein. These findings expand our knowledge of the SADS-CoV replication mechanism and provide additional antiviral strategies for controlling SADS-CoV.

Pathogens doi: 10.3390/pathogens15070751

Authors: Yan Yu Mei Liu Ying Zhang Xinyue Shen Jianmei Li Qin Yang Mei Xue Yabin Dai

Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS) are regarded as the most clinically and economically important avian Mycoplasma species, posing significant challenges to the poultry industry worldwide. In this study, MG and MS antibodies were detected by ELISA in 857 unvaccinated Chinese native chickens from 21 breeder flocks on four multi-age farms. The overall seropositive rates were 86.3% for MG and 98.4% for MS. Offspring from an infected breeder flock were reared under different environmental conditions, and serological monitoring was conducted over a 68-week production cycle. The antibody titers and seropositive rates of both mycoplasmas in birds reared in an isolation room were significantly lower than those in birds reared on the farm after maternal antibodies vanished. By adopting strict biosecurity measures, MG and MS infections were effectively controlled in an offspring flock derived from infected breeders, as evidenced by a substantial decrease in seropositive rates. The results suggested that MG and MS infections were prevalent in the breeder flocks of Chinese native chickens. Horizontal rather than vertical infection was the primary cause of high flock infection levels under field conditions. Strict biosecurity measures could be adopted to initially control MG and MS infections in offspring from the breeder flocks with high infection levels.

Pathogens doi: 10.3390/pathogens15070750

Authors: Célio Alfredo Lurdes Delgado Lúcel Fernandes Omar Manito Mavilingue Ilídio Filipe Manuel António Castro Helder Cortes

Background: Pigs serve as reservoir hosts for various gastrointestinal parasites, posing significant implications for livestock productivity and public health. Data on gastrointestinal parasites in pigs in Mozambique are scarce, while previous studies have focused mainly on porcine cysticercosis. This study aimed to assess the prevalence and diversity of gastrointestinal parasites in pigs of southern Mozambique. Methods: A cross-sectional epidemiological study was conducted with 339 fecal samples, corresponding to the same number of pigs, one sample per animal. Ritchie concentration and Ziehl-Neelsen staining methods were used to detect parasites. Results: An overall prevalence of 88.5% (300/339; 95% CI: 85.1–91.9) was obtained. Eight distinct parasites were identified, with Coccidia exhibiting the highest prevalence at 64.3% (218/339), Strongyle-type eggs at 32.5% (109/339), Cryptosporidium spp. at 26.6% (90/339), Entamoeba spp. at 25.1% (85/339), Balantioides coli at 22.7% (77/339), Ascaris suum at 17.4% (59/339), Giardia spp. at 14.2% (48/339), and Trichuris spp. at 5.6% (19/339). None of the variables analyzed showed a significant association with the infection of pigs (p > 0.05). Conclusions: This is the first large-scale, multi-parasitic epidemiological investigation of gastrointestinal parasites in pigs across southern Mozambique. The detection of Giardia spp., Cryptosporidium spp., Balantioides coli, and Entamoeba spp. underscores the need for molecular methods to clarify the zoonotic potential of these parasites in the region.

Pathogens doi: 10.3390/pathogens15070749

Authors: Jesús Humberto Reyna-Fuentes Pablo González-Alanis Zeferino Blanco-Martínez Flaviano Benavides-González Ana Lucía Urbizu-González María de la Luz Vázquez-Sauceda Mirelly Venecia Mireles-Villanueva

The emergence of multidrug-resistant (MDR) bacteria has increased the need for alternative antimicrobial agents from natural sources. Propolis, a resinous bee product produced by honey bees (Apis mellifera) from plant resins, is rich in bioactive compounds with recognized antimicrobial and antioxidant properties. This study evaluated the phytochemical composition, antioxidant capacity, and antibacterial activity of propolis extracts obtained using ethanol, methanol, acetone, and water against clinically relevant MDR bacterial isolates. Total phenolic content was determined using the Folin–Ciocalteu assay, total flavonoid content by the aluminum chloride colorimetric method, and condensed tannin content by the butanol–HCl assay. Antioxidant activity was assessed using the 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 2,2-diphenyl-1-picrylhydrazyl (DPPH), and ferric reducing antioxidant power (FRAP) assays. Antibacterial activity was evaluated through minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), MBC/MIC ratio, biofilm formation inhibition, and bacterial growth inhibition assays against MDR isolates of Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Enterococcus faecium, and Enterobacter cloacae. Significant differences among extraction solvents were observed for all phytochemical and antioxidant parameters (p < 0.0001). Ethanolic extracts exhibited the highest concentrations of total phenolics, flavonoids, and condensed tannins, as well as the greatest antioxidant activity. MIC values ranged from 3.13 to 25 mg/mL, whereas MBC values ranged from 6.25 to >50 mg/mL depending on bacterial species and extraction solvent. E. coli and E. faecium were the most susceptible isolates, while P. aeruginosa and E. cloacae showed greater tolerance. Most MBC/MIC ratios indicated bactericidal activity. Biofilm inhibition was significantly affected by bacterial strain and extraction solvent (p < 0.001), with ethanolic extracts producing the highest inhibition percentages. These findings demonstrate that ethanolic propolis extracts exhibit strong antioxidant activity, as well as bacteriostatic, bactericidal, and biofilm-formation-inhibitory effects, supporting their potential as complementary antimicrobial agents within a One Health framework.

Pathogens doi: 10.3390/pathogens15070748

Authors: Hande Berk Cam Leyla Kilinc Hasan Avci Hakan Soylu Tugrul Cakir Derya Seyman Filiz Kizilates Nefise Oztoprak Ismail Ustunel

Central venous catheters are a major risk factor for Candida albicans vascular infections, which remain challenging to manage. Although antifungal therapy is standard, the host pathways shaping vascular responses—particularly the Notch signaling pathway (NSP)—are not well characterized in this context. In addition, the potential influence of the prostaglandin pathway on vascular NSP-related responses during infection remains unclear. In this study, a rat model of central venous catheter-associated C. albicans infection was used to evaluate microbiological outcomes and vascular NSP-related protein expression. Immunohistochemical analyses were performed to assess Candida immunostaining alongside the expression of Notch receptors (Notch1–3) and ligands (DLL1/4, Jagged1/2) in vascular tissues. Experimental groups included sham, infected control, antifungal-treated (fluconazole, caspofungin, liposomal amphotericin B), and prostaglandin pathway-intervention groups (sulprostone and sulprostone followed by indomethacin). C. albicans infection was associated with higher vascular NSP-related protein expression compared with sham animals. Antifungal-treated groups showed lower NSP-related protein expression, while fungicidal agents were associated with absence of fungal growth in catheter and kidney cultures. In the sulprostone–indomethacin-treated group, NSP-related protein expression levels were lower than those in the sulprostone-treated group despite persistent fungal burden. In conclusion, central venous catheter-associated C. albicans infection was associated with altered vascular NSP-related protein expression. Differences in NSP-related protein expression patterns were observed across antifungal- and prostaglandin pathway-intervention groups. These findings are descriptive and do not allow causal inference but may provide a basis for future studies exploring the role of NSP in vascular responses to C. albicans infection.

Pathogens doi: 10.3390/pathogens15070747

Authors: Sveinung Wergeland Sørbye Bente Marie Falang Mona Antonsen Elin Richardsen

HPV DNA-positive women with ASC-US/LSIL cytology constitute a heterogeneous triage group. We evaluated genotype-specific HPV E6/E7 mRNA testing with PreTect HPV-Proofer’7 in a population-based cohort from Tromsø and compared the findings descriptively with a previously published cohort from Bodø. This retrospective quality-assurance study included 1006 HPV DNA-positive women with ASC-US/LSIL cytology screened between 2019 and 2024, with linkage to regional pathology records through 31 December 2025. The assay detects E6/E7 mRNA from HPV16, 18, 31, 33, 45, 52, and 58. ASC-US/LSIL accounted for 42.5% of linked HPV DNA-positive women in Tromsø compared with 22.3% in Bodø. Among the mRNA-tested ASC-US/LSIL cohorts, mRNA positivity was similar (40.8% vs. 44.6%). In Tromsø, CIN2+ was recorded in 26.1% of mRNA-positive and 7.9% of mRNA-negative women (RR 3.31, 95% CI 2.41–4.55), whereas CIN3+ was recorded in 3.9% and 1.0%, respectively (RR 3.88, 95% CI 1.53–9.82). For CIN2+, sensitivity was 69.5%, specificity 64.4%, PPV 26.1%, and NPV 92.1%. Among HPV16/18 DNA-positive women, CIN2+ risk was 42.7% in mRNA-positive and 13.4% in mRNA-negative women. Simulated mRNA-guided referral reduced immediate colposcopy referrals by 59.2%. Genotype-specific mRNA testing provided clinically relevant risk stratification, with directionally similar patterns being observed in the Tromsø and Bodø cohorts; however, mRNA-negative women require structured surveillance.

Pathogens doi: 10.3390/pathogens15070746

Authors: Vy Dinh Bao Tran Dong-Hyuk Jeong

Toxoplasma gondii is a zoonotic protozoan transmitted by environmentally persistent oocysts and by tissue cysts in infected prey or meat. Although wild cervids and wild canids are increasingly used as wildlife sentinels, their complementary ecological roles in integrated T. gondii surveillance have not been comprehensively synthesized. This structured narrative review compares infection evidence in five wild cervid species and three wild canid species to examine how feeding ecology shapes exposure and to assess their complementary value in wildlife surveillance. Peer-reviewed literature published between 2000 and 2026 was retrieved from PubMed, Scopus, ScienceDirect, and Google Scholar. Studies reporting evidence of T. gondii exposure or infection in wild cervids or wild canids were included, with serological evidence evaluated separately from molecular or histological detection. Cervids showed geographically variable exposure consistent with ingestion of oocysts from contaminated vegetation, soil, and water, supporting their use as sentinels of environmental contamination. Wild canids often showed higher reported seropositivity, although direct comparisons were limited by assay, sampling, and demographic heterogeneity. Their predatory, scavenging, and omnivorous diets allow access to both environmental oocysts and tissue cysts. Cervids and canids should therefore be treated as complementary rather than interchangeable indicators: cervids primarily reflect environmental exposure, whereas canids integrate environmental and trophic transmission. This review provides an ecological framework to support integrated One Health wildlife surveillance by combining environmental and trophic indicators for improved risk assessment and food-safety planning.

Pathogens doi: 10.3390/pathogens15070745

Authors: Catarina Freitas José Eduardo Pereira Eliana Costa Olga Alves Gilberto Igrejas Patrícia Poeta Vanessa Silva

Coagulase-negative staphylococci (CoNSs) are major opportunistic pathogens in healthcare settings, particularly affecting immunocompromised patients and those with indwelling medical devices. Their growing antimicrobial resistance and ability to form biofilms present significant therapeutic challenges. This study analyzed 148 clinical CoNS isolates to determine species distribution, antimicrobial resistance, resistance genes, and biofilm production. Staphylococcus epidermidis was the most prevalent species (49.3%), followed by Staphylococcus hominis (27.0%) and Staphylococcus capitis (6.8%). Penicillin (83.8%), erythromycin (72.3%), and cefoxitin (66.2%) showed the highest resistance rates. Notably, 60.8% of isolates were resistant to fusidic acid and 45.3% to clindamycin, with inducible resistance in 12.1%. Among aminoglycosides, tobramycin resistance (45.3%) was most frequent. Resistance to ciprofloxacin and trimethoprim–sulfamethoxazole reached 45.9%, while mupirocin resistance was 17.6%. Among isolates resistant to penicillin and/or cefoxitin (n = 128), the mecA gene was detected in 69.5%; the mecC gene was absent. Only 38.5%% of mupirocin-resistant isolates carried the mupA gene. Trimethoprim resistance was mainly associated with dfrA (60.3%) and dfrG (14.7%). Biofilm super-producers accounted for 64.9% of isolates and non-producers for 7.4%, with no significant link between biofilm formation and antibiotic resistance. These findings reinforce the clinical relevance of multidrug-resistant, mupirocin-resistant, and biofilm-forming CoNSs, underscoring the need for improved surveillance and infection control.

Pathogens doi: 10.3390/pathogens15070744

Authors: Sara Valdes José Miguel Saavedra Eugenia Jerez Elida Lebtun Roxana Vargas Pabla Barra Jorge R. Toledo Jaime Romero Harold Oliva Pedro Ilardi

Tenacibaculum maritimum is a major etiological agent of tenacibaculosis in marine fish and represents an increasing concern for Chilean salmon aquaculture; however, updated information on the phenotypic and molecular diversity of circulating isolates is limited. This study characterized 40 isolates recovered from Atlantic salmon (Salmo salar), rainbow trout (Oncorhynchus mykiss), and red cusk eel (Genypterus chilensis) obtained from outbreaks between 2020 and 2024. Isolates were analyzed using biochemical and phenotypic assays, multiplex PCR targeting the O-antigen gene cluster (O-AGC), REP-PCR-based genetic fingerprinting, and experimental bath challenges in Atlantic salmon. Phenotypic characterization revealed species-consistent traits but variable gelatin and starch hydrolysis and differences in seawater tolerance. O-AGC typing identified four molecular serotypes (1-0, 3-1, 3-2, and 4-0), with serotypes 1-0 and 3-2 detected for the first time in Chilean salmonids. Genetic fingerprinting distinguished previously described profiles and two novel REP patterns (REP6 and REP7), indicating additional genomic heterogeneity within dominant serotypes. Virulence assays showed cumulative mortality ranging from 0% to 100%, with serotype 3-1 isolates generally associated with higher mortality and serotype 4-0 displaying broad intra-serotype variability. These findings document substantial phenotypic, antigenic, and genetic diversity among Chilean T. maritimum isolates and provide epidemiologically relevant information for disease surveillance and vaccine design in salmon aquaculture.

Pathogens doi: 10.3390/pathogens15070743

Authors: Umberto Rolla Fabio Persico Giovanbattista Guadagnini Antonio Caleffi Annalisa Scollo

Lawsonia intracellularis, the etiologic agent of porcine proliferative enteropathy (PPE), represents a growing challenge for modern swine production systems. The infection often occurs in a subclinical form and, together with limited awareness of its true production impact, may delay diagnosis and the timely adoption of control measures. In our opinion, this under-recognition of subclinical infection represents the greatest barrier to successful PPE control. Disease management is further complicated by frequent co-infections, limitations of currently available diagnostic tools, and the progressive reduction in routine antimicrobial use, which has revealed the true burden of PPE under commercial conditions. We therefore argue that vaccination should no longer be considered a stand-alone preventive measure, but rather the cornerstone of an integrated, farm-specific management strategy combining accurate diagnostic interpretation, optimized production flows, biosecurity, nutritional management, and continuous veterinary–farmer collaboration. Drawing on both published evidence and the long-term field experience of the authors, this Opinion discusses the key challenges limiting effective PPE control and proposes a practical framework to improve disease recognition, prevention, and sustainable management under commercial production conditions.

Pathogens doi: 10.3390/pathogens15070742

Authors: Sergey V. Shcherbinin Daria A. Bykovtseva Natalia S. Bardina Andrey V. Varkentin

Leptospirosis remains a significant worldwide zoonotic threat, with domestic dogs playing a pivotal role in pathogen circulation across anthropogenic landscapes. This study aimed to assess the spatial distribution of ecological niches of pathogenic Leptospira spp. and conduct epizootic risk mapping within the Russian Federation. A two-stage modeling approach based on the maximum entropy algorithm (MaxEnt) was deployed. At the first stage, the abiotic model revealed the fundamental survival niche of the pathogen, identifying precipitation (700–1200 mm) and temperature (7–12 °C) as critical determinants. The accuracy of this model was AUC = 0.908 ± 0.007. At the second stage, the biotic model achieved a predictive accuracy of AUC = 0.980 ± 0.003, demonstrating that dog population density was the primary driver of risk, exhibiting a distinct threshold pattern (200–300 animals/km2). Furthermore, while rodent density remains the foundational ecological reservoir capable of independent transmission, the presence of additional domestic reservoirs, including livestock and dog populations, synergistically amplifies the transmission pathways, shaping the highest-risk zones within shared environments. These findings indicate distinct spatial risk boundaries within shared human–animal landscapes, suggesting that domestic dogs function as effective indicators for proactive zoonotic monitoring under the “One Health” framework.

Pathogens doi: 10.3390/pathogens15070741

Authors: Nevzat Kılınç Murat Dikilitaş Canan Can Avinash Mishra

Zymoseptoria tritici (Desm. Quaedvlieg & Crous), known as the wheat leaf spot disease agent, is a highly virulent fungus that induces blotch and necrosis on leaves. Although it is known to cause severe infections under humid conditions, recent observations suggest that it can also infect wheat leaves under drought and high-temperature conditions, possibly influenced by global warming. Recent findings showed that Z. tritici could easily tolerate various abiotic stresses, including drought, water stress, salinity, and temperature. It has been evident that the fungus can tolerate pesticide stress, as indicated by the increased frequency and number of pesticide applications throughout the growing season. Under stress conditions, the fungi, unlike crop plants, could easily tolerate stress by rapidly modifying gene expression and reducing spore production and mycelial growth without downregulating major biochemical components that play significant roles in pathogenicity and virulence. Z. tritici can accumulate melanin under stress conditions; therefore, an increase in pathogenicity under drought or salinity stress is not unexpected. Recent studies have shown that the pathogenicity of the fungus is increasing, and more virulent, toxin-producing pathogens might emerge in the future. Since drought and high-temperature stresses significantly affect crop plants, the adaptation of pathogenic microorganisms to these conditions could be inevitable if abiotic stress persists. Under these circumstances, the crop loss would be more pronounced. A critical aspect of this process is the assessment of DNA integrity in both wheat and the pathogen under drought stress conditions. The organism that better maintains DNA integrity is considered to exhibit greater drought tolerance. Therefore, our main target should be DNA health when developing or breeding new wheat varieties, considering double- or even multiple-stress conditions. We should finally state that we are very optimistic about generating highly stress-tolerant wheat varieties via metabolomic and proteomic approaches without compromising quality. However, the impact and combination of stress factors are becoming increasingly complex.

Pathogens doi: 10.3390/pathogens15070740

Authors: Andrea Marino Emmanuele Venanzi Rullo Giuseppe Pipitone Alessandro Giorgio Geremia Andrea De Vito Antonio Albanese Nicholas Geremia Alessandro Franzò Federica Cosentino Paolo Italia Maria Elena Ciuppa Andrea Buzzi Laura Santoro Chiara Iaria Giordano Madeddu Carmelo Iacobello Rosa Manuele Fabrizio Pulvirenti Bruno Cacopardo Giuseppe Nunnari

Background: Long-acting lipoglycopeptides (LALs), including dalbavancin and oritavancin, may facilitate outpatient-oriented management of Gram-positive infections by reducing the need for prolonged hospitalization and daily intravenous therapy. However, real-world evidence on their use in day hospital pathways, particularly for complex off-label infections, remains limited. This study evaluated the clinical effectiveness, safety, and economic impact of LAL-based day hospital management, comparing in-label acute bacterial skin and skin-structure infections (ABSSSI) with complex off-label indications. Methods: This retrospective, multicenter observational study included 160 adult patients treated with dalbavancin and/or oritavancin in a day hospital setting across nine Italian centers between January 2020 and December 2025. Indications were classified as in-label ABSSSI or off-label infections, including osteomyelitis, prosthetic joint infection, spondylodiscitis, endocarditis, septic arthritis, and prosthetic cardiac infection. The primary endpoint was clinical success at final follow-up. Secondary endpoints included adverse drug events, readmissions, inflammatory marker response, and a budget impact cost-offset analysis based on avoided inpatient bed-days. Results: Overall, 54.4% of patients received LALs for off-label indications, mainly osteomyelitis, prosthetic joint infection, and spondylodiscitis. Clinical outcome was available for 159 patients, with an overall success rate of 86.8% (138/159). Success rates were 90.4% for in-label ABSSSI and 83.7% for off-label indications, with no statistically significant difference between groups (p = 0.247). In exploratory analyses, monotherapy was associated with lower failure odds; this most likely reflects confounding by indication and should not be interpreted as a treatment effect. Inflammatory markers significantly decreased from baseline to end of therapy. Adverse drug events were uncommon (3.3%), mild, and did not lead to treatment discontinuation. In a scenario-based cost-offset model (no matched inpatient comparator), the strategy corresponded to 1107–2214 avoided inpatient bed-days; estimated net savings ranged from approximate cost-neutrality under conservative assumptions (€11,976) to €676,176 under maximum assumptions, and one-way sensitivity analysis showed a possible net loss at low inpatient daily-cost values—underscoring that these are modeled rather than observed savings. Conclusions: In this uncontrolled, retrospective cohort, LAL-based day hospital management was feasible and associated with favorable observed clinical outcomes, an acceptable safety profile, and a potential for cost offset in selected patients with Gram-positive infections, including complex off-label indications. Because no matched inpatient or conventional OPAT comparator was included, these findings should be regarded as hypothesis-generating and cannot establish comparative efficacy, superiority, or actual cost savings.

Pathogens doi: 10.3390/pathogens15070739

Authors: Chao Yan Xuanfeng Liu Yujie Chen An Su Xue Ren Tingting Zhang Jing Yuan

This study aimed to use quantitative phosphoproteomics to explore phosphorylation characteristics in Klebsiella pneumoniae (Kpn)-induced liver abscess (KPLA) formation and liver injury. The Kpn strain LA-Kpn006 was phenotypically and genotypically characterized. A murine model of KPLA was established via intragastric inoculation. Phosphoproteomics and bioinformatic analyses were conducted to identify and quantify phosphosites and phosphoproteins. LA-Kpn006 displayed a hypermucoviscous phenotype, serotype K1, ST23 genotype, and harbored six major virulence genes. Inoculation induced liver colonization and typical histopathological abscess lesions. We quantified 3017 phosphoproteins covering 12,798 phosphosites (dominated by serine phosphorylation); 1723 proteins were upregulated and 425 downregulated. Bioinformatic analyses revealed remodeling in metabolism, stress response, signal transduction, and cytoskeleton organization. Upregulated proteins converged on fatty acid elongation, inositol phosphate metabolism, and the tricarboxylic acid cycle; downregulated proteins were enriched in PI3K–Akt, IL-17 signaling, and T-cell differentiation. Protein–protein interaction network analysis identified 10 key proteins (Src, Rac1, Actb, Hsp90aa1, Hsp90ab1, Egfr, Rps6, Pik3CA, Itgb1, and Fyn) that mediate inflammatory signaling, cytoskeleton remodeling, and immune infiltration. Dysregulated phosphorylation networks drive pathological metabolic adaptation, suppressed immune homeostasis, and cytoskeletal disorganization, collectively facilitating KPLA progression. The identified hub proteins and pathways represent high-value mechanistic targets and candidate therapeutic vulnerabilities for KPLA.

Pathogens doi: 10.3390/pathogens15070738

Authors: Lucas R. G. Crosby Md Fahim Khan Larry S. McDaniel Lance E. Keller

Streptococcus pneumoniae is a leading cause of respiratory infections and is often treated with β-lactam antibiotics despite frequent resistance. The β-lactam resistance is through mutations in penicillin-binding proteins (PBPs), but the impact of PBP regulation remains poorly understood. Here, we investigate the role of gene regulation by the oligopeptide-binding protein aliD on β-lactam susceptibility. aliD-expressing strains exhibit elevated minimum inhibitory concentrations (MICs) to the β-lactam antibiotics amoxicillin and cefdinir compared to isogenic aliD mutants. In contrast, aliD had minimal effects on susceptibility to vancomycin, suggesting a mechanism specific to β-lactam antibiotics. We demonstrate that aliD increases clpL expression with or without antibiotics present. Inducible expression of clpL resulted in stepwise increases in amoxicillin MIC, demonstrating that elevated clpL expression directly contributes to decreased β-lactam susceptibility. Furthermore, we show that increased clpL expression enhances PBP2x surface exposure in a dose-dependent manner. Additionally, aliD-expressing strains exhibited significantly increased cell wall cross-linking relative to aliD mutants. Together, these findings identify a novel regulatory pathway in which aliD enhances clpL expression, promotes PBP2x surface exposure, and increases peptidoglycan cross-linking, ultimately reducing β-lactam susceptibility. This work expands current understanding of pneumococcal antibiotic resistance and suggests that peptide sensing through oligopeptide transport systems may influence antibiotic susceptibility in host-specific environments.

Pathogens doi: 10.3390/pathogens15070737

Authors: Florian Plaku Ilir Kusi Esmeralda Dushku Anastasia Paraskeva Virginia Giantzi Erinda Lika Fatbardh Sallaku Theofilos Papadopoulos Elena Tsavea Charalampos Kotzamanidis

This study presents the first comprehensive molecular characterization of Escherichia coli producing extended-spectrum beta-lactamases (ESBL-Ec) in surface waters in Albania, focusing on the Shkumbini river. Antimicrobial resistance (AMR) in aquatic ecosystems poses a significant threat to public health, yet data from Albania remain scarce. Thirty water samples were collected from six locations near Elbasan between September 2022 and February 2024. Following the WHO Tricycle protocol, 52 ESBL-Ec isolates were recovered and characterized for antimicrobial susceptibility, biofilm formation, resistance genotypes and clonal relatedness via pulsed-field gel electrophoresis (PFGE). ESBL-Ec was detected in 80% of the samples analyzed, with 94.2% of the isolates classified as multidrug-resistant (MDR). High resistance frequencies were observed for ampicillin (98.1%) and cefotaxime (86.5%), while 7.7% of the isolates displayed colistin resistance associated with the mcr-3 gene. The blaCTX-M-1 genotype was the most prevalent (57.7%), and almost half of the isolates harbored multiple ESBL genes. Phylogroup A (46.2%) predominated, followed by the high-risk extraintestinal lineages B2 (23.1%) and D (11.5%). PFGE revealed high genetic heterogeneity, with 51 distinct pulsotypes indicating multiple sources of contamination, such as untreated municipal, agricultural and industrial waste. Additionally, 55.8% of the isolates were capable of forming biofilms. These results highlight the critical role of the Shkumbini river as a reservoir for highly resistant pathogens and emphasize the urgent need for integrated environmental surveillance and improved wastewater management in Albania.

Pathogens doi: 10.3390/pathogens15070736

Authors: Andrea Tedesco Pietro Sponga Giulia Bertoli Andrea Matucci Graziana Da Rold Federica Obber Lucia Moro Chiara Piubelli Francesca Perandin Concetta Castilletti Fabio Formenti Cristina Mazzi Salvatore Scarso Dora Buonfrate Federico Gobbi

Background: Data on tick-borne diseases (TBDs) in Italy are fragmented. This study aimed to describe tick-borne pathogens (TBPs) and TBDs observed in a hospital in Italy and to assess the correlation between TBPs and human disease. Methods: A prospective observational study was conducted in a single hospital between March 2024 and January 2025. Inclusion criteria were: individuals ≥ 8 years of age and consent to study participation. Data and blood samples were collected at baseline (T0), and 12–18 weeks later (T1). Ticks collected at T0 were pooled and analyzed by real-time PCR. Serological and molecular tests were used for TBD diagnosis. Results: Ninety-three individuals (median age 39 years; 52.7% female) were enrolled. The pathogens most frequently detected in the tick pools were Rickettsia spp. (15.1%) and Borrelia spp. (7.5%). Sixteen participants (17.2%) developed TBD-compatible symptoms, but TBD was confirmed in only four cases: Lyme borreliosis (n = 2), scalp eschar and neck lymphadenopathy after tick bite (SENLAT, n = 1), and tick-borne encephalitis (TBE, n = 1). Concordance between TBP detection and the corresponding TBD development in humans was observed in only two cases (2.1%). Conclusions: The low concordance between pathogens detected in participant-level tick pools and confirmed human tick-borne disease supports the limited clinical value of tick testing in this setting.

Pathogens doi: 10.3390/pathogens15070735

Authors: Rana H. M. Hussien Alexandra M. Kortsinoglou Martyn J. Wood Vassili N. Kouvelis Wanissa Mellikeche Mustapha Touray Babalwa Tembeni Mazen Alzain Faisal Alotaibi Islam S. Sobhy Zack Saud E. Joel Loveridge Daniel C. Eastwood Tariq M. Butt

Entomopathogenic fungi (EPF) are well established as biological control agents, but their emerging role as endophytes reveals a broader and more powerful function in crop protection. By colonizing plant tissues, endophytic entomopathogenic fungi (EEPF) create a dynamic tripartite interaction between plants, fungi, and herbivores, enabling systemic, plant-mediated pest suppression. This review synthesizes current knowledge on the behavioral and ecological responses of herbivorous arthropods to EEPF-colonized plants, with an emphasis on the mechanisms and implications for integrated pest management (IPM). Growing evidence indicates that EEPF consistently modify herbivore behavior and performance across diverse crops and insect taxa. Colonization frequently alters feeding, host selection, and oviposition, often deterring pests, although mediated responses may vary among fungal species, host plants, insect taxa, and environmental conditions. These responses are driven by EEPF-induced changes in plant chemistry, including shifts in volatile organic compounds (VOCs) and defensive metabolites. In parallel, EEPF impair insect fitness by delaying development, reducing survival, and lowering fecundity, thereby suppressing pest populations. These plant-mediated and behavioral changes extend to multitrophic interactions, potentially affecting associations with natural enemies and the transmission efficiency of some insect vectors of plant viruses. Despite rapid progress, critical gaps remain in resolving the mechanistic basis of these interactions and their stability under field conditions. Advancing the application of EEPF will require integrated approaches combining microbial ecology, chemical ecology, and insect behavioral biology. Harnessing these interactions offers a compelling pathway to reduce reliance on synthetic pesticides while enhancing the resilience and sustainability of agricultural systems.

Pathogens doi: 10.3390/pathogens15070734

Authors: Maria Katsiari Charikleia Nikolaou Kalliopi Theodoridou Eleftheria Palla Athanasios Tsakris Georgia Vrioni

Introduction: Candidemia is the most significant invasive fungal disease in critically ill patients. As a shift towards non-albicans Candida (NAC) species has been observed in recent years, differentiation among Candida species is essential for appropriate therapeutic management and improved prognosis. This study evaluated trends in the epidemiology of candidemia, species-specific characteristics, treatment practices, and mortality in a multidisciplinary Greek intensive care unit (ICU). Materials and Methods: In this single-center retrospective observational study, 90 ICU patients with candidemia were evaluated. Clinical characteristics, infection-related factors, treatment practices, and outcomes were compared according to isolated Candida species and patient survival. Results: C. parapsilosis accounted for the majority of isolates (33.7%; 41.9% fluconazole-resistant), followed by C. albicans (31.5%) and Candidozyma auris (23.9%). No significant differences in co-morbidities, origin of candidemia, or treatment-related factors were identified. However, NAC bloodstream infections occurred significantly later during ICU hospitalization, were associated with lower disease severity, and were preceded by nearly five-fold higher exposure to antifungal agents before candidemia onset. Overall, the ICU mortality rate was 53.3%. No significant differences were observed among species-specific mortality rates. Neither prompt initiation of antifungal therapy nor the antifungal class administered was associated with improved survival. Conclusions: NAC species predominated among candidemia episodes in critically ill patients, with C. parapsilosis emerging as the most frequent isolate and exhibiting substantial fluconazole resistance. Mortality remained high and did not differ significantly according to species. Treatment timing and antifungal class were not associated with outcome, highlighting the need for continued surveillance and optimized management strategies in the ICU.

Pathogens doi: 10.3390/pathogens15070733

Authors: Xuanheng Tai Yiying Zhang Huijie Yang Wei Zou

The immunological paradigm is undergoing profound shifts. Classic “self-nonself” recognition remains foundational, but microbiome research has expanded immunological models toward homeostasis, tolerance, and host–microbe co-regulation. Within this framework, gut microbiota act as the core drivers of the development and calibration of the host immune system. This review examines host–microbiome immune interactions across species from a macroevolutionary perspective. We use C. elegans, mouse models, and humans as comparative systems representing different levels of immune complexity and translational relevance. These include Caenorhabditis elegans (primitive innate immunity via cytosolic surveillance), mouse models (microbiota-driven shaping of adaptive immunity and homeostatic trade-offs), and the human system (modern lifestyle-induced “evolutionary mismatch” and inflammatory diseases). The immune system has evolved significantly over time. It has transitioned from independent cellular integrity monitoring to a complex network that is heavily reliant on external microbial cues. This potential integration of developmental signals from commensal microbes enhances environmental adaptability but may also increase host susceptibility to inflammatory disorders under modern ecological shifts. Deconstructing this cross-species interaction blueprint offers significant theoretical value. Furthermore, it provides the guiding logic for developing next-generation precision microbiome therapies that target the holobiont, such as engineered microbial consortia and personalized postbiotic interventions.

Pathogens doi: 10.3390/pathogens15070732

Authors: Suhana Chattopadhyay Leena Malayil Emmanuel F. Mongodin Amy R. Sapkota

Tobacco products (e.g., cigarettes, little cigars) harbor diverse bacterial communities and long-term tobacco use alters the oral microbiome, potentially leading to oral disease. However, no studies have evaluated the immediate changes in the oral bacterial microbiota that could occur after using a new tobacco product. To address this knowledge gap, buccal swab and saliva samples were collected from forty cigarette smokers before and after a single use of a little cigar product on two separate visits. Total DNA was extracted from a total of 320 samples. The 16S rRNA gene was amplified from these samples and sequenced on the Illumina HiSeq to characterize the bacterial microbiota. Oral bacterial diversity was not significantly different between pre- and post-smoking samples. However, post-smoking buccal samples were enriched with Delftia, Leptotrichia, Pseudomonas and Stenotrophomonas (genera that can include opportunistic pathogens) and post-smoking saliva samples were enriched with Catonella when compared to the pre-smoking samples. In summary, single use of a little cigar product does not immediately impact overall oral bacterial diversity among cigarette smokers; however, post-smoking oral samples may have a higher relative abundance of some bacterial genera. Hence, smoking a new tobacco product could potentially alter the relative abundance of some bacterial types within the oral cavity of smokers, highlighting a potential microbiological pathway through which tobacco use could contribute to oral disease risk.

Pathogens doi: 10.3390/pathogens15070731

Authors: Jing Liao Xingxin Liu Jingjing Luo Jiaji Ling Liting Liang Ziyi Yan Wenjing Wu Wei Zhou Yongmei Jiang

The escalating problem of bacterial drug resistance poses a severe threat to global public health, with Acinetobacter baumannii (A. baumannii) exhibiting particularly high resistance rates that are closely linked to its Quorum Sensing (QS) system. This narrative review synthesizes current knowledge on the A. baumannii QS system, focusing on its essential role in mediating antimicrobial resistance, as well as the latest progress in developing QS inhibitors. Key findings indicate that the A. baumannii QS system enhances bacterial survival by promoting biofilm formation, and regulating the expression of efflux pump and resistance genes. However, significant translational challenges remain, including the risk of inducing resistance, the poor bioavailability and suboptimal pharmacokinetic properties, and their reduced efficacy in complex biological systems. In conclusion, while QS inhibitors represent a promising therapeutic target, overcoming current developmental bottlenecks requires combining them with traditional antibiotics, exploring novel drug delivery strategies, and integrating cutting-edge tools to accelerate drug discovery and clinical application.

Pathogens doi: 10.3390/pathogens15070730

Authors: Alberto Rizzo Davide Moschese Federica Salari Luca Carsana Samuel Lazzarin Andrea Cavallo Chiara Fusetti Loriana Morelli Francesco Caruso Alessandra Lombardi Andrea Giacomelli Manuela Nebuloni Alberto Dolci Andrea Gori

The clinical relevance of concurrent vaccine-preventable anal HPV genotype infections, particularly for short-term repeated detection and cytologic outcomes in screened high-risk populations, remains uncertain. We performed a single-centre retrospective study of 145 adults with at least one 9-valent vaccine-preventable anal HPV genotype at baseline and repeat HPV DNA testing and cytology approximately 12 months later. Baseline infections were classified as single- or multiple-genotype infections. Outcomes included clearance of all baseline vaccine-preventable genotypes, genotype-specific repeated detection, detection of new vaccine-preventable genotypes, complete HPV negativity, and cytologic regression or progression. At baseline, 70 participants (48.3%) had single and 75 (51.7%) had multiple genotype infections; most were male (95.2%) and people living with HIV (88.3%), reflecting a highly selected anal HPV-screened cohort. Multiple infections were associated with more frequent abnormal baseline cytology (69.3% vs. 45.7%). At follow-up, clearance of all baseline vaccine-preventable genotypes was more common after single than multiple infections (34.3% vs. 12.0%). Complete HPV negativity was also more frequent after single infection (24.3% vs. 4.0%), although this stringent endpoint is intrinsically more difficult to achieve in individuals with multiple baseline infections. New vaccine-preventable genotype detection did not differ significantly between groups. Genotype-specific repeated detection was generally similar between groups, except for HPV58, a finding based on small subgroup numbers. Cytologic regression was more frequent after single infection, but not statistically significant. Multiple vaccine-preventable anal HPV genotype infections were associated with greater baseline cytologic abnormality and lower clearance of baseline vaccine-preventable genotypes at 12 months. Given the retrospective design, selected population, limited event counts, and residual confounding, these findings should be interpreted as hypothesis-generating and do not prove impaired biological clearance.

Pathogens doi: 10.3390/pathogens15070729

Authors: Gabriella Locsmándi Enikő Kádár-Hürkecz Zsuzsa Kienle Katalin Tárnoki-Boross Krisztina Sima Tímea Erdősi László Egyed

One hundred and ninety free-ranging rat individuals from dozens of sampling areas (towns, farms, villages) were investigated for Leptospira infections. From the renal tissues (kidneys and urinary bladders), DNA was extracted, and the samples were screened by an lfb1-specific PCR assay. The PCR products were sequenced. One hundred and three (54.2%) of the samples proved to be positive. All detected Leptospires belonged to the pathogenic phylogenetic cluster, and only one species was detected: all 103 positive samples belonged to L. interrogans lfb1 species group 1. In parallel, 353 human samples (urine, anticoagulated blood, tissues) from 232 patients over the period 2022–2025 submitted with suspected Leptospira infections were tested by commercial multiplex real-time PCR kits. Twenty-nine (8%) positive samples were found, which were retested by lfb1-specific PCR. The quality of 20 PCR products was sufficient for sequencing, representing 14 individual patients. Among the 14 positive patients we identified two Leptospira species: L. kirschneri lfb1 species group 6 in one case and L. interrogans in 13 cases (one lfb1 species group 3 imported case, 12 lfb1 species group 2). Comparison of the lfb1 sequences obtained from rats (lfb1: 1) and human cases (lfb1: 2, 1: 3, 1: 6) indicated that, although rat populations maintain the pathogen in high prevalence, they could not be the sources of the identified human Leptospira infections.

Pathogens doi: 10.3390/pathogens15070728

Authors: Daniel A. Nwaubani Rakshya Baral Tamunobelema Solomon Mustafa Ali Tania Moharrery Samendra P. Sherchan

This study established long-term wastewater surveillance of Aichi virus (AiV) in Maryland. AiV, a member of the Kobuvirus genus associated with acute gastroenteritis, has established itself as an integral marker for wastewater-based monitoring; however, two key research questions remain unaddressed for the Baltimore metropolitan area: (1) whether AiV is consistently detectable in municipal wastewater throughout the year, and (2) whether its concentrations exhibit a measurable seasonal pattern. To address these hypotheses, influent samples were collected on a weekly basis from WWTP-A and WWTP-B from January to December 2023 (with grab sampling conducted at WWTP-A and automated collection deployed for the influent sampling of the water treatment plant B). All samples (n = 51) were subjected to PEG 8000 concentration, RNA extraction, cDNA synthesis, and RT-qPCR quantification. We observed AiV RNA in 94.12% of the samples from both facilities (25/51 at WWTP-A and 23/51 at WWTP-B) with concentrations that ranged from 2.5 to 3.63 log10 gc/L and a seasonal pattern showing consistent declines: loads for WWTP-A declining from winter (3.58 log10 gc/L) to fall (2.56) and for WWTP-B from winter (3.28 log10 gc/L) to fall (2.31). The year-round constant AiV presence provides a strong basis for its use as a stable viral marker within wastewater-based epidemiology efforts.

Pathogens doi: 10.3390/pathogens15070727

Authors: Ignacio Hernan Pineda Etcheber Cheryld Mutel Gonzalez Javiera Antonia Bascuñan Maiz Antonia Cesped Astete Mauricio Soto Vasquez

Infective endocarditis (IE) is a severe pathology with recent changes in its epidemiological profile, characterised by older patients with more comorbidities. The objective of this study is to describe the clinical and microbiological characteristics, as well as potential factors associated with mortality, of patients with IE in a tertiary academic centre. Material and Methods: Descriptive, retrospective, and observational study of patients over 18 years of age with a confirmed diagnosis of IE, conducted between 2021 and 2023 at the Dr Hernán Henríquez Aravena Hospital in Temuco, Chile. Biodemographic variables, risk factors, microbiology, echocardiographic findings, and complications were analysed using descriptive statistics and logistic regression models. Results: 119 patients were included (average age 60 years; 65.5% male; 28.5% rural). The most frequent risk factors were arterial hypertension (55%) and diabetes mellitus (29%). 18% were on haemodialysis (HD). Microbiological isolation was achieved in 78.1% of cases, with Streptococcus gallolyticus the most frequent isolate (16.8%), followed by Staphylococcus aureus (15.1%) and coagulase-negative Staphylococcus (15.1%). Complications were present in 69% of cases, mainly emboli (43%) and septic shock (23%)—59.6% required surgery. Global mortality was 44.5%, with a decreasing annual trend (from 58% in 2021 to 33% in 2023). Comorbidities and complications independently associated with mortality were chronic renal failure on HD (OR 5.76; p = 0.001), heart failure (OR 3.13; p = 0.025), and septic shock (OR 3.31; p = 0.016). Conclusions: IE in this centre presents an aggressive profile and a high burden of comorbidities. The prevalence of S. gallolyticus is a notable regional observation not reported in similar populations. Mortality remains high, with an improving trend.

Pathogens doi: 10.3390/pathogens15070726

Authors: Marisia Madrigal-Solis Mirjam Kaestli Mark Mayo Celeste Woerle Ella M. Meumann Bart J. Currie

Background: Melioidosis correlates strongly with rainfall, and there is substantial diversity in climate between melioidosis-endemic locations. The Northern Territory of Australia epitomises the “wet/dry” tropics, with a prolonged dry season from May to October. We analysed dry season cases of melioidosis during 35 consecutive years and compared these with wet season cases. We aimed to provide insights into how dry season cases of melioidosis may occur in this region and explore non-rainfall exposures that are usually not considered in the wet season. Methods: Case epidemiological and clinical data were extracted from the Darwin Prospective Melioidosis Study. Weather parameters, including daily rainfall, were analysed using generalised additive models and conditional logistic regressions to assess associations between dry season cases and preceding rainfall. Results: Of 1520 melioidosis cases between 1989 and 2024, there were 325 (21%) in the dry season. While the well-recognised clinical diversity of melioidosis was also seen amongst dry season cases, pneumonia was proportionally less common and cutaneous melioidosis was more common than in the wet season. A total of 23% of dry season patients had no identified clinical risk factors for melioidosis, compared to 14% in the wet season. Mortality was 8% in the dry season and 11% in the wet season. There was a range of plausible explanations for many of the dry season cases, including unseasonal rainfall prior to infection. Infections in urban settings were notable, with anthropogenic factors such as irrigation and construction resulting in persistence of Burkholderia pseudomallei in the environment during the dry season. A total of 3% of cases remained unexplained. Conclusions: Not all dry season cases are explained by infection occurring the previous wet season or by unseasonal rainfall in the dry. Identification of cases in the dry season support the need for year-round prevention strategies during potential exposure to contaminated water or soil. Further prospective studies are needed to better define the infecting events resulting in melioidosis, especially in the dry season. These studies should include timely history taking from the case and their family and selected environmental sampling for B. pseudomallei.

Pathogens doi: 10.3390/pathogens15070725

Authors: Ewelina Truszkowska Małgorzata Salamonowicz-Bodzioch Jowita Frączkiewicz Krzysztof Kałwak Filip Pierlejewski Małgorzata Nowak Maciej Zdunek Wojciech Młynarski Krzysztof Czyżewski Kamila Jaremek Oliwia Grochowska Patrycja Zalas-Więcek Katarzyna Derwich Weronika Stolpa Karolina Baranowska Agnieszka Mizia-Malarz Olga Gryniewicz-Kwiatkowska Magdalena Łukszo Bożenna Dembowska-Bagińska Ewa Bień Ninela Irga-Jaworska Jan Styczyński Olga Zając-Spychała

Serratia species are Gram-negative pathogens responsible for a wide range of nosocomial infections. This multicenter nationwide retrospective study aimed to describe the epidemiology, clinical characteristics, antimicrobial susceptibility, and outcomes of Serratia infections in pediatric oncology patients and hematopoietic stem cell transplantation (HSCT) recipients in Poland between 2012 and 2023. A total of 36 Serratia infection episodes were identified in patients under 20 years of age, including 30 cases (83.3%) in the oncological (OHD) group and six (16.7%) among HSCT recipients. The median age was 4.30 years. The most common underlying diseases were acute lymphoblastic leukemia (36.1%) and central nervous system tumors (16.7%). Bloodstream infections predominated in OHD patients (33.3%), whereas urinary tract infections were most frequent in HSCT recipients (83.3%). S. marcescens was the most commonly isolated species. More than half of isolates (53.3%) showed antimicrobial resistance, with extended-spectrum β-lactamase (ESBL)-producing strains in 26.7% and AmpC β-lactamase-producing strains in 13.3%. Multidrug resistance occurred in 30%. Treatment most often included amikacin, piperacillin/tazobactam, and carbapenems. Five deaths occurred in the OHD group and one in the HSCT group, none directly related to Serratia infection. Although uncommon, Serratia infections remain clinically relevant due to their high antimicrobial resistance, underscoring the need for antimicrobial stewardship.

Pathogens doi: 10.3390/pathogens15070724

Authors: Elena De Carolis Terenzio Cosio Carlotta Magrì Marialaura Del Mondo Riccardo Torelli Flora Marzia Liotti Paola Bernaschi Tiziana D’ Inzeo Giovanni Vento Maurizio Sanguinetti

Background: Rare fungal infections may represent under-recognized causes of healthcare-associated sepsis, particularly when caused by emerging or difficult-to-identify pathogens. We aimed to characterize Candida viswanathii isolates recovered in the setting of panel-negative candidemia and to assess the contribution of an integrated diagnostic workflow. Methods: We investigated seven C. viswanathii isolates overall, including three recovered at our institution from blood, urine, and bronchoalveolar lavage of a NICU patient, as well as four bloodstream isolates from a second pediatric center included for comparison. Isolates were analyzed by culture and microscopy, three MALDI-TOF MS platforms, internal transcribed spacer sequencing, Fourier transform infrared (FTIR) spectroscopy and antifungal susceptibility testing. Results: C. viswanathii was repeatedly recovered from blood, urine and bronchoalveolar lavage, while the FilmArray BCID2 panel remained negative. All MALDI-TOF MS systems with updated databases correctly identified the yeast at the species level; identification was confirmed by sequencing. Fourier transform infrared analysis showed clustering of clinical isolates and clearly separated C. viswanathii from related Candida species. All isolates exhibited low MICs to echinocandins and amphotericin B as well as moderately elevated fluconazole MICs (2–4 mg/L). Conclusion: This study supports the use of explicit diagnostic algorithms for rare fungal pathogens in yeast-positive, syndromic panel-negative blood cultures. In this setting, updated MALDI-TOF MS libraries and FTIR spectroscopy may provide useful adjunctive support for the recognition and phenotypic discrimination of atypical yeasts within an integrated laboratory workflow.

Pathogens doi: 10.3390/pathogens15070723

Authors: Gorana Veinović Sanja Ćakić Darko Mihaljica Ratko Sukara Aleksandra Stefanović Elizabeta Ristanović Eva Ružić-Sabljić Snežana Tomanović

Serbia is characterised by the high prevalence and diversity of Borrelia burgdorferi sensu lato strains in ticks, but the reported incidence of Lyme borreliosis is much lower than that in regions with a similar prevalence of Borrelia in ticks. To evaluate the susceptibility and viability profiles of Borrelia strains circulating in local enzootic cycles, we analysed their response to human complement in vitro using short-term incubation periods. A total of 31 strains, comprising 27 Serbian tick isolates, two tick isolates from Spain, one tick isolate from Portugal, and one human skin isolate from Portugal (12 Borrelia afzelii, 12 Borrelia lusitaniae, three Borrelia bavariensis, two Borrelia garinii, and two Borrelia valaisiana), were analysed using serum susceptibility testing. The motility of spirochetes was assessed one and three hours after incubation with normal human serum (NHS) and heat-inactivated serum (HIS) used as a control. The absence of a statistically significant decrease in strain motility after incubation with NHS compared with HIS indicated the resistance of the strains. All tested B. afzelii and B. bavariensis strains and one B. valaisiana strain were serum-resistant, whereas both B. garinii strains were serum-susceptible. Based on statistical analysis, all B. lusitaniae strains were classified as serum-susceptible; however, two local tick isolates and the human reference strain demonstrated distinct biological heterogeneity, maintaining significantly higher residual motility after short-term exposure to human serum, which underscores strain-specific and geographic variations in complement tolerance.

Pathogens doi: 10.3390/pathogens15070722

Authors: Joanna Kulisz Zbigniew Zając Aneta Woźniak Sara Moutailler Angélique Foucault-Simonin Alejandro Cabezas-Cruz

Ixodes ricinus is the principal vector of numerous tick-borne pathogens (TBPs) in Europe and is typically associated with forest sites that provide favorable microclimatic conditions. However, this species may also occur in meadow ecosystems, which are generally regarded as suboptimal environments and remain insufficiently studied from an epidemiological perspective. The aim of this study was to determine and compare the prevalence of Borrelia spp. and Rickettsia spp. in adult I. ricinus occurring in urban and rural meadow sites in eastern Poland. Ticks collected between June 2023 and May 2024 were screened for Borrelia spp. and Rickettsia spp. using high-throughput microfluidic real-time PCR targeting the 23S rRNA and ITS regions, respectively. The DNA of Borrelia spp. was detected in 14.8% of ticks from the urban site and 10.7% from the rural site, whereas Rickettsia spp. were detected in 5.6% and 8.9% of specimens, respectively. No significant differences in pathogen prevalence were observed between sites. The results confirmed the presence of Borrelia spp. and Rickettsia spp. in adult I. ricinus collected in urban and rural meadow sites.

Pathogens doi: 10.3390/pathogens15070721

Authors: Na Young Hong Minyu Qin Min A Lim Youkyoung Kim Sook Hee Park Sung Jun Park Hyun Jun Kang Byeong Ryeon Kim Ji Hyuk Park Seok Ju Yoo Kwan Lee

Vaccination remains an effective intervention for reducing coronavirus disease 2019-related deaths, while the population-level evidence regarding the association between vaccination dose and mortality remains limited. This study aimed to investigate the association between vaccination status, independent risk factors, and COVID-19 mortality, using population-based surveillance data from Gyeongsangbuk-do, South Korea. A population-based retrospective cohort study was conducted using 698,537 confirmed cases from Gyeongsangbuk-do, between January 2021 and June 2022, including 1008 deaths. Univariable and multivariable logistic regression analyses were performed to identify factors associated with mortality. Additional analyses were conducted among adults aged ≥ 65 years. The overall case fatality rate was 0.14%, increasing to 0.88% among older adults. Older age (≥65 years; OR = 87.262, 95% CI: 67.265–114.472) and underlying disease (OR = 20.394, 95% CI: 17.136–24.270) were strongly associated with mortality. After adjustment for sex, age, and underlying disease, unvaccinated individuals had substantially higher odds of death than vaccinated individuals (aOR = 7.897, 95% CI: 6.903–9.034). Booster vaccination (≥3 doses) was associated with markedly reduced mortality in both the overall population (aOR = 0.094, 95% CI: 0.081–0.109) and older adults (aOR = 0.119, 95% CI: 0.102–0.139). Advanced age, underlying diseases, and lack of vaccination were significant factors associated with COVID-19 mortality during the mass vaccination period. These findings support continued efforts to improve booster vaccination coverage among older adults and individuals with underlying diseases.

Pathogens doi: 10.3390/pathogens15070720

Authors: Katarzyna W. Pancer Magdalena Rosińska Gerhard Dobler Daniel Rabczenko Agnieszka Kołakowska-Kulesza Beata Gad Anna Poznańska Piotr Grabarczyk

TBEV is a major cause of viral central nervous system infections in Europe, with heterogeneous geographical distribution and substantial underdiagnosis in low-incidence regions. This study aimed to evaluate the validity of regional TBE risk classification in Poland by combining surveillance-based incidence data with serological markers of TBEV exposure. Plasma samples from 5541 blood donors residing in nine regions were tested by anti-TBEV IgG ELISA, followed by confirmatory VNT, IFA, and anti-NS1 IgG ELISA to differentiate infection-induced from vaccine-induced antibodies. Regions were classified based on average TBE incidence registered in surveillance systems in 2015–2019. Overall, 272 (4.9%) donors were positive in TBEV IgG ELISA and 177 (3.2%) also in a confirmatory assay. Seroprevalence expressed by markers consistent with past TBEV infection (anti-NS1 IgG) was estimated at 0.13% (95% CI 0.05–0.26%), ranging from 0.0% to 0.33% by voivodeship, whereas vaccine-induced immunity accounted for the majority of samples with detected specific antibodies (3.1%). Surprisingly, seroprevalence in a highly affected region (0.29%) was at the same level as in one that was less affected (0.33%). Moreover, all except one seropositive donor lived in urban areas. In four out of nine voivodeships, no anti-NS1 TBEV IgG was detected. By utilising precise laboratory algorithms, we demonstrated a much lower seroprevalence that estimated from prior research relying on screening tests. In addition, we confirmed low vaccination coverage. Integrating sero-epidemiological data with surveillance systems may improve risk assessment and inform targeted prevention strategies.

Pathogens doi: 10.3390/pathogens15070719

Authors: Alís Hassan Salman Lara Nazareth Afonso Ana Jullia Alves Guimarães Andressa Alves Soares Paula Pereira da Matta Ari Sérgio de Oliveira Lemos Patrícia de Almeida Machado Juliana da Trindade Granato

The Pan American Health Organization (PAHO) recognizes ethnicity as a structural determinant of health, yet ethnic–racial analyses of neglected tropical diseases remain scarce. This study investigated cumulative notification and mortality rates of American cutaneous leishmaniasis (ACL) in Brazil from 2016 to 2025 according to the official Brazilian race/color categories. Data were obtained from the Notifiable Diseases Information System (SINAN/DATASUS), and temporal and spatial patterns were analyzed. ACL showed marked geographic heterogeneity, with the highest cumulative notification rates concentrated in the North and Central-West regions. Indigenous and Asian populations presented the highest cumulative notification rates, whereas mortality rates were highest among Indigenous populations and remained elevated among Brown populations in several settings. Spatial clusters were concentrated mainly in the Brazilian Amazon, reinforcing the persistence of territorial inequalities in ACL distribution. Overall, the findings reveal substantial race/color and geographic disparities in both ACL occurrence and mortality in Brazil, underscoring the need for more equitable surveillance, prevention, and control strategies.

Pathogens doi: 10.3390/pathogens15070718

Authors: Yurong Song Ming Cheng Xiaoling Li Ying Wang Yulong Zong Jing Li Guozhong Chen Shiyong Chen Yuan Cao

Vibrio vulnificus (V. vulnificus) is a type of bacterium commonly found in estuarine environments. It can cause necrotizing wound infections and sepsis, both of which are associated with high mortality rates. Protein lysine acetylation is a widespread post-translational modification (PTM) of proteins, and it participates in numerous cellular processes, including regulation, in bacteria. However, the finer landscape of lysine acetylation in V. vulnificus remains unexplored. In this study, acetylated proteins with low cellular abundance were enriched from V. vulnificus MO6-24/O using anti-acetyl-lysine immunoprecipitation and identified using LC-MS/MS, and the acetylation was further confirmed by Western blot analysis. We mapped 2035 lysine acetylation sites to 841 proteins, accounting for approximately 18.5% of the entire protein sequence of V. vulnificus MO6-24/O. Comprehensive bioinformatic characterization of the acetylome indicated that lysine acetylation is associated with metabolic regulation, particularly targeting enzymes regulating carbon metabolic functions and biosynthesis. In addition, sequence motif analysis identified two conserved patterns surrounding acetylated lysines: enrichment of lysine or arginine residues at the +4/+5 positions, and a preference for tyrosine, histidine, or phenylalanine residues at the −1/+1 positions. Furthermore, analysis of the protein–protein interaction network indicated that lysine acetylation influences numerous molecular interactions among proteins. Collectively, this acetylome investigation establishes a foundation for future studies aimed at elucidating physiological functions of protein lysine acetylation in V. vulnificus.

Pathogens doi: 10.3390/pathogens15070717

Authors: Maryna Golovchenko Lucie Krätzerová Heather MacTavish Vett Lloyd Natalie Rudenko

Disseminated human Lyme borreliosis is primarily associated with invasive spirochetes from the Borrelia burgdorferi sensu lato complex. Host–Borrelia interactions have been studied in a diverse range of vertebrate reservoirs. But despite the key role of the complement system in innate immunity, comparative studies evaluating the susceptibility of individual Borrelia species to human complement-mediated killing are limited. Using serum sensitivity assays, we analyzed complement-mediated killing of 10 B. burgdorferi s.l. genospecies in healthy individuals of different ages and sexes. The tested genospecies showed markedly different sensitivities to human complement. Statistical clustering divided the genospecies into three distinct groups based on whether their sensitivity to human complement was high, medium, or low. Complement resistance did not correlate with the recognized pathogenicity of the genospecies; species with unclear pathogenic potential exhibited resistance levels comparable to the major species causing human Lyme borreliosis, B. burgdorferi sensu stricto, B. afzelii and B. garinii. Females generally showed reduced complement-mediated killing compared to males. In addition, complement activity tended to decline with age, identifying host age and biological sex as important factors influencing the human innate immune response against Borrelia. Our findings suggest that the ability to evade human complement may be more widespread among Borrelia species than previously assumed. These results support an emerging model in which complement-mediated selection may shape host associations and transmission success without serving as an absolute predictor of Borrelia virulence.

Pathogens doi: 10.3390/pathogens15070716

Authors: Kiramage Chathuranga W. A. Gayan Chathuranga Tania F. de Koning-Ward Jong-Soo Lee

Ubiquitination is a post-translational modification that governs various facets of eukaryotic biology, including protein stability, signaling, and immune regulation. The modification process is mediated by a coordinated enzymatic cascade, in which E3 ubiquitin ligases confer substrate specificity and determine the functional outcome of ubiquitin attachment. In the case of a virus infection, host cellular signaling networks undergo major ubiquitin-dependent changes to protect the host cell, including remodeling of cellular organelles, coordination of innate immunity, and reprogramming of metabolic pathways to prevent virus replication. African swine fever virus (ASFV) has evolved numerous strategies to counteract or evade these responses, thereby manipulating host defenses and promoting its replication. By modulating ubiquitination-dependent host cellular functions, the virus can regulate key immune signaling factors, suppress interferon production, and interfere with inflammatory pathways. These actions not only antagonize antiviral defenses but also remodel cellular homeostasis to favor infection. The important interplay between host defense and viral manipulation underscores the versatility of the ubiquitin system as a battleground in ASFV infection. In this review, we discussed mechanistic insights into how ASFV subverts ubiquitin pathways during host–virus interactions. This comprehensive knowledge might be beneficial for pharmaceutical exploration of host E3 ligase-dependent anti-ASFV treatment.

Pathogens doi: 10.3390/pathogens15070711

Authors: Lekshmi K. Edison Subhashinie Kariyawasam

Foodborne pathogens remain a major public health challenge, particularly in the context of antimicrobial resistance and persistent contamination across animal, food-processing, and retail environments. This review examines bacteriophages as precision antimicrobials for controlling major foodborne bacteria, including Salmonella, Campylobacter, Shiga toxin-producing Escherichia coli (STEC), Listeria monocytogenes, and Vibrio spp., and summarizes the biological basis of phage-mediated control: strictly lytic life cycles, receptor-specific adsorption, direct bacterial killing, biofilm disruption, and resistance-associated fitness trade-offs. It further discusses pre-harvest, post-harvest, and processing-environment applications, with emphasis on matrix-dependent efficacy, delivery strategies, commercial products, and regulatory status. While bacteriophages offer high specificity and may help preserve the native microbiome, their integration into multi-hurdle food-safety systems require careful validation because their performance is influenced by narrow host ranges, bacterial resistance, food-matrix effects, formulation constraints, and regulatory complexity and scale-up challenges. Broader implementation will require rationally designed phage-cocktails, thorough genomic safety screening, matrix-specific validation studies, scalable manufacturing processes, and continuous monitoring for post-application resistance. Overall, bacteriophages should be viewed as promising but context-dependent adjuncts to validated food-safety and One Health frameworks, rather than stand-alone solution for reducing foodborne pathogen burdens.

Pathogens doi: 10.3390/pathogens15070714

Authors: Asuman Akar

Background: Healthcare-associated bloodstream infections remain a significant cause of morbidity and mortality in hospitalized children, particularly in intensive care settings. Carbapenem-resistant Gram-negative bacterial (CR-GNB) bloodstream infections and fungemia may present with overlapping clinical features. This can complicate empirical treatment decisions in resource-limited settings. This study evaluated baseline clinical and laboratory characteristics associated with CR-GNB bloodstream infections and fungemia among high-risk pediatric patients. Methods: This retrospective observational cohort study included pediatric patients aged 0–18 years who were evaluated at the time of clinical deterioration and blood culture collection for suspected healthcare-associated bloodstream infection before empirical antifungal therapy initiation for the index episode. Patients who subsequently received empirical antifungal therapy between May 2023 and September 2025 were retrospectively screened. Of the 240 screened patients, 103 met the inclusion criteria and were classified into CR-GNB (n = 56) and fungemia (n = 47) groups based on blood culture results. Clinical, laboratory, and microbiological data were analyzed using univariate and multivariable statistical methods. Results: Observed 90-day all-cause mortality was higher in the CR-GNB group than in the fungemia group (50.0% vs. 29.8%, p = 0.038). Central venous catheter use was more frequent (91.1% vs. 48.9%, p = 0.006), and platelet counts were lower (median: 120 × 109/L vs. 259 × 109/L, p = 0.011) in patients with CR-GNB bloodstream infections. In multivariable analysis, thrombocytopenia (OR: 4.22, 95% CI: 1.35–13.17; p = 0.013) and central venous catheter use (OR: 5.53, 95%: CI 1.89–16.26; p = 0.002) were independently associated with CR-GNB bloodstream infections. The model showed moderate discrimination (AUC = 0.786). Conclusion: In this selected high-risk cohort, thrombocytopenia and central venous catheter use were associated with CR-GNB bloodstream infections. Observed mortality was higher in the CR-GNB group, but this finding should be interpreted with caution as adjusted mortality analysis and standardized severity assessment were not performed. These findings are hypothesis-generating and require validation in larger prospective studies before guiding empirical treatment decisions.

Pathogens doi: 10.3390/pathogens15070715

Authors: Madison R. Stevens Kristin Hamaker Ian Gereg Matthew Shaub Lane Potts Erica A. Miller Taylor C. Chan Madeline Vile Andrew Di Salvo Kevin D. Niedringhaus

Rabies is a universally fatal viral disease that affects a wide variety of mammalian species, including carnivores and herbivores. While testing and confirmed reports in wild carnivores are common, comparatively less is known about this disease in wild herbivores. Between 2022 and 2024, nine white-tailed deer (Odocoileus virginianus) in Pennsylvania, USA were diagnosed with rabies. Eight (89%) presented with erratic neurologic signs and one was found dead. Evidence of chronic self-mutilation characterized by hair loss and thickened, hardened, and hyperpigmented skin was present on the head in seven cases (78%) and/or body or legs in two cases (22%), while two (22%) did not have any evidence of external lesions. Microscopically, lesions were like those in other mammalian species, with variably severe inflammation and abundant cytoplasmic inclusion bodies with associated antigen immunoreactivity. This case series highlights a common and characteristic gross lesion that can prompt caution and testing for rabies in deer and further expands understanding of this disease in an uncommonly diagnosed species.

Pathogens doi: 10.3390/pathogens15070713

Authors: Fatima Hisam Anisha Reddy Konakalla Eranda Berisha Maria del Carmen Chacon Castro Spandan Mukherjee Claire Wang Benjamin R. Sheirbon Tracie Delgado Erica L. Sanchez

Gammaherpesviruses (GHVs) represent a global clinical burden as the causative agents of Kaposi’s sarcoma and mononucleosis, among other diseases. Kaposi’s sarcoma-associated herpesvirus (KSHV) and Epstein–Barr virus (EBV) are the most studied human GHVs, and murine gammaherpesvirus 68 (MHV-68) is a recognized experimental model. GHVs are defined by their modulation of the host cell to establish lifelong latent infections and increase host dysregulation during periodic reactivation. Due to their ubiquitous changes in host cells, systems-level techniques are well-suited to study GHV infections at all stages of the central dogma: genomics, transcriptomics, and proteomics. Furthermore, metabolomics can reveal the final metabolic changes across numerous host cellular pathways. This review assesses the current knowledge on GHV infections gained through omics techniques. We also identify gaps and propose future directions, including the development of new therapeutic strategies. Early omics techniques have characterized large swaths of infection for EBV, KSHV, and MHV-68, revealing conserved genes, homologous transcripts, and proteins. Modern omics techniques have enabled higher-resolution studies, yielding insights into heterogeneity in viral-host gene, transcript, and protein modulation strategies across geographical populations, viral subtypes, inter- and intra-patient infections, and latent and lytic states. The metabolome during GHV infections remains the least understood, but current studies have identified essential modulations of nucleotide, amino acid, and lipid synthesis by EBV, KSHV, and MHV-68. Importantly, the application of integrative omics methods to GHV infections remains a promising direction of study as the increased resolution of modern techniques meets the need for greater understanding of differences in each GHV infection.

Pathogens doi: 10.3390/pathogens15070712

Authors: Hakan Çarpar Ömer Faruk Coşkun

During the 2024 growing season, black carrot (Daucus carota subsp. sativus) plants showing symptoms consistent with phytoplasma infection, including leaf chlorosis, reduced leaf size, witches’ broom, excessive fibrous root formation, multiple lateral taproots, and floral phyllody, were observed in production fields in Hatay Province, Türkiye. To identify the associated phytoplasma, 23 symptomatic plants and two asymptomatic control plants were analysed using PCR-based molecular detection, sequencing, BLASTn comparison, and phylogenetic analyses of the 16S rRNA, secA, tuf, and imp gene regions. The SAP11 gene was also screened as an additional virulence-associated molecular marker, but no functional characterization was performed. All symptomatic samples yielded amplicons of the expected sizes for the targeted loci, whereas no amplification was obtained from asymptomatic controls. Sequence comparisons revealed >99% nucleotide identity with members of the peanut witches’ broom group, and multilocus phylogenetic analyses consistently placed the black carrot phytoplasma isolate within the 16SrII-D subgroup. To our knowledge, this study provides the first documented evidence of a 16SrII-D phytoplasma associated with black carrot in Türkiye. The finding is relevant for plant pathology and plant protection because it indicates the occurrence of a phytoplasma lineage with potential epidemiological importance in an economically important vegetable production area. These results provide a basis for future studies on disease distribution, insect vectors, alternative host plants, epidemiology, and management strategies in black carrot production systems.

Pathogens doi: 10.3390/pathogens15070709

Authors: Eglė Rudaitytė-Lukošienė Liuda Kutkienė Saulius Švažas Dalius Butkauskas Petras Prakas

Sarcocystis spp. are apicomplexan parasites that form sarcocysts mainly in the muscles or central nervous system of intermediate hosts and sporocysts in the intestines of definitive hosts. Three species, Sarcocystis falcatula, Sarcocystis calchasi and Sarcocystis halieti, are potentially pathogenic to their intermediate hosts. Over the past two decades, we have examined 72 thrushes across four species (redwing (Turdus iliacus), common blackbird (Turdus merula), song thrush (Turdus philomelos), and fieldfare (Turdus pilaris)) for sarcocysts to better understand their role as intermediate hosts. Sarcocysts were detected in 28 individuals (38.9%). Most sarcocysts observed by light microscopy were of a single morphological type consistent with Sarcocystis turdusi. A molecular analysis of ITS1 sequences confirmed the presence of S. turdusi in the common blackbird, song thrush, and fieldfare, establishing the latter two bird species as new intermediate hosts. In contrast, cox1 was not a sufficiently variable locus for species differentiation. Additionally, a single sarcocyst with a smooth cyst wall, distinct from S. turdusi, was detected in a common blackbird and identified as S. halieti based on ITS1 sequence analysis. This atypical host record represents an isolated finding in a long-term dataset. Further sampling is required to confirm its epidemiological significance.

Pathogens doi: 10.3390/pathogens15070710

Authors: Ramchander Nadipelly Mohamed Selim Jagathiswaran Radhakrishnan Gun Temeeyasen Eric Nelson Travis J. Clement Naveen Duhan Sunil K. Mor

Porcine reproductive and respiratory syndrome (PRRS) is a serious disease that causes reproductive failure and late abortion in breeder farms, as well as severe respiratory manifestations in pigs of all ages. In the United States, the swine industry has been affected by PRRSV-2, particularly due to control challenges and the virus’s ongoing evolution. In this study, we conducted a genetic characterization of the 642 PRRSV-2 ORF5 sequences generated from diagnostic submissions at the Animal Disease Research and Diagnostic Laboratory (ADRDL) at South Dakota State University (SDSU). These sequences were generated over a 3-year period from 2023 through 2025, with 208 (32.4%), 130 (20.2%), and 304 (47.4%) sequences in 2023, 2024, and 2025, respectively. These sequences were from swine diagnostic submissions from ten states in the Midwest, including Minnesota (MN; 350/642, 54.5%), South Dakota (SD; 182/642, 28.3%), and Iowa (IA; 73/642, 11.4%), collectively accounting for 94.2% of submitted sequences. The genetic typing revealed that the L1C sub-lineage was the most frequently detected among the submitted sequences, accounting for around 60%, with L1C.5 as the major contributor (371/642). Other sub-lineages, such as L5A, L1A, L1D, and L1H, were also detected, but in smaller numbers. More than 80% of the detected sequences were classified as wild-type or wild-like based on ORF5 identity to commercial vaccine reference strains. Interestingly, this study documented increased detection of the recently reported L1C.5.32 variant, which was the most frequently detected variant in 2025 (123 detections), particularly in Minnesota. Moreover, L1C.5.34, L1C.5.36, L1C.3.25, and L1C.2 variants were also detected, and the L1H.18 variant showed increased detection, especially in 2025. In conclusion, this study provides genetic characterization of PRRSV-2 ORF5 sequences from Midwestern U.S. diagnostic submissions (a key U.S. swine production region), submitted to the SDSU ADRDL.

Pathogens doi: 10.3390/pathogens15070708

Authors: Jhon Omar Palomino-Tenorio Obert Marín-Sánchez Jimmy Ango-Bedriñana Ruy D. Chacón Homero Ango-Aguilar

Hepatitis B virus (HBV) infection remains a major public-health challenge in Peru, particularly in historically hyperendemic Amazonian and Andean regions; however, the structure, evolution, and equity of national HBV research have not been systematically evaluated. We conducted a PRISMA-informed bibliometric analysis of all peer-reviewed and theses on HBV in Peru published between 1988 and 2023 using Scopus, Google Scholar, and the Peruvian National Repository (RENATI). Bibliometric indicators, collaboration networks, thematic structure, and temporal thematic evolution were analyzed in R using bibliometrix- and network-based approaches. The final corpus comprised 232 documents, with a marked increase in production after 2005 and a publication peak in 2018. Scientific output was strongly concentrated in Lima-based institutions, while several departments historically associated with HBV endemicity exhibited minimal or absent research production. Nearly half of the corpus corresponded to undergraduate and postgraduate theses. Thematic analyses revealed persistent predominance of epidemiology, seroprevalence, and vaccination-related research, whereas molecular virology, therapeutics, and translational research remained peripheral or poorly represented. International collaboration was markedly limited. Overall, Peruvian HBV research has expanded quantitatively but remains geographically centralized and shows only limited correspondence with the contemporary geographic distribution of HBV incidence, while also remaining only partially aligned with the contemporary global HBV research frontier. These findings provide an evidence-based framework to guide research-priority setting, territorial equity policies, and strategic investment in infectious disease research capacity in Peru. Moreover, the weak association observed between scientific production and departmental HBV incidence suggests that factors beyond contemporary epidemiological burden contribute to the current distribution of research activity in Peru, highlighting a critical but often overlooked dimension of health inequity in low- and middle-income countries (LMIC) research systems.

Pathogens doi: 10.3390/pathogens15070707

Authors: Alexander Martens Markus Schauer Mohamad Motevalli Susanne Mair Brigitte König

Background: Public fitness facilities are high-contact environments that facilitate microbial transfer via shared surfaces; however, temporal dynamics and device-specific contamination patterns remain insufficiently characterized. Methods: A repeated-measures observational study was conducted in a fitness facility over five consecutive weekdays (Monday to Friday). A total of 180 surface samples were collected from 12 gym devices, each sampled three times daily (morning, noon, and evening). Surface-associated cultivable bacteria were recovered using culture-based methods followed by MALDI-TOF MS identification. Ecological metrics, including species richness and Shannon diversity, were calculated, and taxa were classified by origin (skin-associated versus environmental). Device-specific contamination profiles were developed using a composite index incorporating pathogen presence, contamination frequency, and persistence. Temporal trends and predictors of contamination were analyzed using mixed-effects regression models. All statistical analyses were performed in R. Results: A total of 248 bacterial isolates were identified, representing 61 species across 32 families, with a predominance of skin-associated taxa (72.2%). Sampling time point was a strong independent predictor of contamination (adjusted OR for noon vs. morning: 7.19; p < 0.001). While overall microbial diversity remained stable across devices (Shannon index, p = 0.44), substantial heterogeneity was observed in pathogen prevalence, multispecies burden, and persistence. The functional trainer and leg extension showed the highest composite risk scores (42.3%), while the ab crunch machine and upper body ergometer demonstrated significantly increasing contamination trends over the sampling period (p < 0.05). Co-occurrence analysis showed nonrandom microbial associations, with the strongest positive links between Micrococcus luteus and Staphylococcus saprophyticus (Φ = 0.76) and Staphylococcus aureus (Φ = 0.61). Conclusions: Gym equipment surfaces harbor predominantly human-associated microbial communities exhibiting dynamic temporal contamination patterns, and on selected devices, increasing the baseline contamination across consecutive cleaning cycles. The findings indicate that contamination patterns on shared fitness equipment are dominated by taxa commonly associated with human skin and support targeted hygiene interventions focused on frequently contacted devices and periods of elevated contamination.

Pathogens doi: 10.3390/pathogens15070706

Authors: Xiuli Zhao Yinling Li Ziyan Wang Zhenzhou Wan Chiyu Zhang

This study aimed to investigate the prevalence of human pegivirus type 1 (HPgV-1) among healthy individuals, type 2 diabetes patients, and cancer patients in Taizhou, Jiangsu, China. A total of 2872 participants, including 2052 healthy individuals, 373 type 2 diabetes patients, and 447 cancer patients, were enrolled at Taizhou Fourth People’s Hospital between 2022 and 2024. Serum samples were collected from each participant, and HPgV-1 RNA was detected using a reverse transcription-polymerase chain reaction (RT-PCR) assay. The positive rates were compared across the three groups. The age range of healthy individuals was 5–91 years, while that of type 2 diabetes and cancer patients was 25–85 years and 34–91 years, respectively. The prevalence of HPgV-1 among healthy individuals was 15.3% (313/2052), which was significantly higher than that in type 2 diabetes patients (11.0%, 41/373, p < 0.05) and cancer patients (9.2%, 41/447, p < 0.001). In healthy individuals, the prevalence of HPgV-1 increased from childhood to young adulthood, peaking in the 19–40 years age group, followed by a decline after 40 years of age, indicating an age-related pattern. Similarly, HPgV-1 load increased from childhood to adulthood. No significant gender differences in HPgV-1 prevalence were observed across the three cohorts. In conclusion, HPgV-1 infection exhibits an age-dependent prevalence, with the lowest rate in children and adolescents and the highest in young adults (19–40 years). The significantly lower prevalence of HPgV-1 in type 2 diabetes and cancer patients raises the intriguing question of whether HPgV-1 infection may play a protective or contributory role in the pathogenesis of diabetes and cancers.

Pathogens doi: 10.3390/pathogens15070705

Authors: Anna J. Harte Elias Mafuru Athumani Ramadhani Tamsyn Derrick Harry Pickering Tara Mtuy Patrick Massae Ehsan Ghasemian Aiweda Malissa Robin L. Bailey David C. W. Mabey Matthew J. Burton Martin. J. Holland

Trachoma, caused by Chlamydia trachomatis (Ct), persists as a major cause of preventable blindness despite the global SAFE strategy. Understanding how Ct genovars and genovariants influence infection dynamics and clinical outcomes is crucial for sustaining elimination efforts and informing vaccine development. A four-year longitudinal study was conducted in a trachoma-endemic region of Tanzania across multiple rounds of mass drug administration (MDA) with azithromycin. Ct infections were genotyped by ompA sequencing to identify genovars and genovariants. Associations between genetic variants, bacterial load, and clinical signs of trachoma were assessed. Following MDA, a shift in Ct genovar prevalence occurred from genovar B to genovar A. Genovar B was associated with more severe clinical signs, including follicles, papillae, and scarring, whereas genovar A infections exhibited higher bacterial loads. Among 121 individuals with recurrent infections, 94% were re-infected with the same genovar, indicating limited protective immunity and incomplete clearance despite MDA coverage exceeding 60%. The genovariants B2, B9, and A2 predominated, with an A → T amino acid substitution in B9 potentially modifying antigenic recognition. Post-MDA, normalized genovariant diversity increased, suggesting ongoing transmission or strain reintroduction. Distinct genovar-associated clinical and immunological patterns underscore the need to elucidate genovar-specific virulence and immune evasion mechanisms. These findings provide key insights for optimizing trachoma control and advancing vaccine development.

Pathogens doi: 10.3390/pathogens15070704

Authors: Catherine A. MacNary Sai Chaitanya Rajendra Gaekwar Alexander Lemenze Ayaan Naik Ritesh Tandon Salwa Ahmed Bobby Brooke Herrera Pooja Jain

Human T-lymphotropic virus type 1 (HTLV-1)-associated myelopathy/tropical spastic paraparesis (HAM/TSP) is a chronic neuroinflammatory disorder often leading to demyelination of the spinal cord. Progression to HAM/TSP is closely associated with the high proviral load and the presence of virally infected CD4+ T cells that release extracellular vesicles (EVs). Exosomes, an EV subtype released by many cell types, transport proteins and nucleic acids that regulate intercellular communication and have been implicated in the progression of cancer and neuroinflammatory diseases. Herein, we have studied the effect of exosomes from HTLV-1 infected cells on the Peripheral Blood Mononuclear Cells (PBMCs) of HAM/TSP patients by single-cell sequencing utilizing innovative Honeycomb technology. We observed a distinct transcriptional response in monocyte populations compared with other immune cell types. Given that monocytes remain understudied in HTLV-1 pathogenesis, these findings highlight a potential role for infection-derived exosomes in shaping monocyte-driven immune dysregulation in HAM/TSP. A total of 41 genes were identified to be differentially expressed in HAM/TSP monocytes treated with exosomes; 28 were upregulated and 13 were downregulated. The most significantly altered genes are involved in chemokine activity and signaling, macrophage differentiation, lipid metabolism, and lysosomal function. Overall, our data suggests that exosome-treated HAM/TSP monocytes undergo immune remodeling that favors cell recruitment, activation, and a shift toward a mixed M1/M2-like phenotype. Such a shift may support viral persistence and chronic inflammation. These findings highlight a potential therapeutic pathway for addressing HTLV-1-induced neuroinflammation by modulating exosome-mediated signaling.

Pathogens doi: 10.3390/pathogens15070703

Authors: Antonia Touloudi Alexios Giannakopoulos Panagiota Tyrnenopoulou Athanasios Siasios Zoi Athanasakopoulou Garyfallenia Tsinopoulou Marina Sofia Vassiliki Spyrou George C. Fthenakis Charalambos Billinis Dimitrios C. Chatzopoulos

Equids, primarily horses, are mostly used for recreational purposes, although in some rural areas they also serve as working animals, maintaining close and frequent contact with humans. Their risk of exposure to vector-borne and zoonotic pathogens can be affected by host-related factors, management practices and environmental conditions. This study aimed to investigate the seroprevalence and associated risk factors for infections by Babesia caballi, Toxoplasma gondii, Coxiella burnetii, and Borrelia burgdorferi sensu lato in equids from Southwestern Greece. A total of 159 equids were tested using commercial serological assays. Weighted prevalence estimates were applied to account for unequal sampling. Associations were assessed using chi-square tests and logistic regression. Ecological niche modelling was employed to evaluate geographic patterns and environmental correlates. Seroprevalence was highest for B. caballi (8.81%), followed by T. gondii (7.55%) and C. burnetii (1.26%). No seropositive animals were detected for B. burgdorferi sensu lato. Ecological niche modelling showed acceptable predictive performance for B. caballi, with BIO14 and BIO6 emerging as the main environmental predictors. In contrast, the T. gondii model exhibited unacceptable predictive performance, and its environmental associations should therefore be interpreted cautiously. Complementary Random Forest analyses yielded comparable environmental rankings but showed higher classification performance for T. gondii than for B. caballi. Overall, the findings contribute to understanding pathogen exposure patterns in equids and underscore the importance of integrating epidemiological and environmental data in surveillance efforts.

Pathogens doi: 10.3390/pathogens15070702

Authors: Bach Xuan Pham Linh Khanh Bui Tawin Inpankaew

Canine vector-borne pathogens (CVBPs) pose a major challenge in shelter medicine, yet data from shelter populations in Vietnam remain unknown. This study determined the prevalence, and risk factors of CVBPs in shelter dogs in northern Vietnam. Blood samples from 300 apparently healthy dogs from three shelters in Hanoi were screened by PCR for Babesia vogeli, Hepatozoon canis, Rickettsia spp., and Mycoplasma spp. Representative positive amplicons underwent Sanger sequencing and BLAST analysis. Sequence analysis showed 96.07–100% identity with reference strains, with phylogenetic trees confirming clustering within B. vogeli, H. canis, M. haemocanis and R. felis clades. Overall, 43.7% (131/300) of dogs were infected with at least one pathogen, with shelter-level prevalence ranging from 38.0 to 52.0%. Single infections accounted for 35.0%, dominated by R. felis (25.7%) and M. haemocanis (24.0%), B. vogeli and H. canis were low (1.3% each). Co-infections were found in 8.7% of dogs, primarily R. felis and M. haemocanis (8.3%). No evaluated host factors (age, sex, breed, body size, housing style) significantly associated with infection (p > 0.05). This study provides the first molecular evidence of canine vector-borne pathogen circulation in Vietnamese shelter dogs, emphasizing the need of ectoparasite control and One Health-oriented surveillance.

Pathogens doi: 10.3390/pathogens15070701

Authors: Holly Yu Amanda R. Mercadante Kate Halsby Alexandra Loew-Baselli Ye Tan Juanita Edwards Frederick J. Angulo Elizabeth Begier Mendwas Dzingina Johan S. Berglund Anna Moniuszko-Malinowska Franc Strle James H. Stark on behalf of the BOLD Study Group on behalf of the BOLD Study Group

Lyme borreliosis (LB) can manifest as a localized infection or, if left untreated, disseminated disease. This analysis used Patient-Reported Outcome tools to assess general health, fatigue, pain, and cognitive function in patients with localized or disseminated LB, compared with age- and site-matched controls without LB. All participants were enrolled in the Burden of Lyme Disease (BOLD) study; LB cases were assessed at 3 time points: the enrollment visit, and two follow-up visits conducted within 10 months of enrollment. Controls were evaluated at 2 time points: the enrollment contact and a follow-up contact 16–18 months post-enrollment. Symptom severity and general health quality between groups were compared using two-tailed Student’s t-tests and multivariate regression analyses. Disseminated LB cases reported greater fatigue and pain than localized LB (disseminated LB fatigue scores: 4.1, 3.7, and 3.5 at Visits 1, 2, and 3, respectively vs. localized LB: 2.9, 3.0, and 2.9; disseminated LB pain scores: 2.5, 1.6, and 1.3 vs. localized LB: 0.9, 0.6, and 0.8) and controls (fatigue score: 3.2; pain score: 1.0). Disseminated LB was also associated with reduced health-related quality of life. The results of this study indicate that health-related quality of life may vary by disease stage and that patients with disseminated disease showed persistent impairment during the acute phase and at 10-month follow-up.

Pathogens doi: 10.3390/pathogens15070700

Authors: Darío S. López-Delgado Mathias S. Renteros-Ramirez Joshua Emmanuel Arteaga-Bolaños Harold E. Vásquez-Ucros Kevin Alexander Burbano-Castro Valentina Reina-Melo Jessica Niebles-Blanco Nancy Calzada-Gonzales Lysien I. Zambrano Valmore Bermudez Alfonso J. Rodriguez-Morales

We sincerely thank Iqhrammullah and Rampengan for their thoughtful and valuable Comment [...]

Pathogens doi: 10.3390/pathogens15070699

Authors: Muhammad Iqhrammullah Derren D. C. H. Rampengan

We read with great interest the systematic review and meta-analysis by López-Delgado et al. [...]

Pathogens doi: 10.3390/pathogens15070698

Authors: Nefer Roberta Gianotto Neftj Ragusa Virginia Deut Chiara Mattivi Marta Cherubini Scarafoni Silvia Dominici Giulia Mazzetti Matteo Sandei Chiara Lo Presti Cenni Manuela Mario Michele Calvo Massimo Berger

Respiratory syncytial virus (RSV) is the leading cause of bronchiolitis and hospitalization in infants worldwide. In 2024, the Piedmont region introduced universal immunoprophylaxis with Nirsevimab for all infants experiencing their first RSV season. We carried out a multicenter retrospective observational study across the three pediatric units of ASL TO4 (Ivrea, Ciriè, Chivasso), comparing bronchiolitis-related hospitalizations during the 2023–2024 season (pre-Nirsevimab) with those from the 2024–2025 season (post-Nirsevimab). The primary outcome was the proportion of RSV-positive hospitalizations. Secondary outcomes included age at admission, need for respiratory support, PICU/NICU transfer, and length of stay. Immunization coverage was assessed using the regional electronic registry. Immunization coverage exceeded 88% across all centers (overall 90.4%). A total of 179 bronchiolitis hospitalizations were recorded (134 pre- vs. 45 post-Nirsevimab). RSV-positive admissions showed a reduction from 70.9% to 55.6% after implementation (OR 0.52; 95% CI 0.24–1.09). Center-specific analyses suggested reductions in Ciriè (OR 2.48; 95% CI 1.41–4.39) and Chivasso (OR 2.28; 95% CI 1.09–4.77), with a similar trend observed in Ivrea. In a supplementary denominator-based analysis restricted to infants younger than 12 months, RSV-related hospitalization incidence decreased from 42.0 to 10.3 per 1000 infants between seasons (OR 4.23; 95% CI 2.64–6.78; p < 0.0001). Disease severity remained unchanged between seasons in terms of respiratory support, length of stay, and PICU/NICU transfers. Age at admission increased significantly during the post-intervention season (mean 118.3 vs. 160.9 days; Welch’s two-sample t-test, p = 0.026). Among 15 immunized infants hospitalized in 2024–2025, 6 were RSV-positive, none required intensive care, and only two needed high-flow nasal cannula (HFNC). Universal Nirsevimab prophylaxis was associated with a trend toward reduction in RSV-related hospitalizations at the aggregate level, although the overall comparison did not reach statistical significance. Center-specific analyses suggested reductions in RSV-positive admissions in some participating units. A supplementary denominator-based analysis among infants younger than 12 months showed a lower incidence of RSV-related hospitalizations during the post-implementation season. No evidence of increased severity among breakthrough cases was observed. High coverage demonstrated the feasibility of implementation and its potential public health value. Continued longitudinal surveillance over additional RSV seasons is essential to better define the durability of protection and long-term epidemiological impact.

Pathogens doi: 10.3390/pathogens15070697

Authors: Vasiliki Rapti Anna-Pelagia Magiorakos Efthymia Giannitsioti Garyfallia Poulakou

Since its first description in the late nineteenth century, the epidemiology of infective endocarditis (IE) has changed considerably. Once primarily affecting younger individuals with structural heart disease, IE is now increasingly encountered in older patients with multiple comorbidities and frequent healthcare exposure. Population ageing, end-stage renal disease (ESRD), immunosuppression, and injection drug use (IDU) have broadened the pool of susceptible hosts. At the same time, the increasing use of prosthetic valves (PVs), cardiac implantable electronic devices (CIEDs), and transcatheter cardiac interventions has reshaped the clinical spectrum of IE. This epidemiological transition has also been accompanied by shifts in microbiological patterns, with a growing predominance of staphylococci and enterococci, as well as marked geographic and socioeconomic variation in disease burden. This review summarizes the contemporary epidemiology of IE, with an emphasis on the host-, healthcare-, and microbiological factors underlying its evolving clinical profile.

Pathogens doi: 10.3390/pathogens15070696

Authors: Jose G. Montoya Stephanie M. Cho Stephanie Smith Carlos A. Gomez Despina G. Contopoulos-Ioannidis

Targeted immunotherapies with biologics, small molecules, and CAR T-cell therapies have revolutionized treatment across autoimmune, chronic inflammatory, oncologic, and transplant-related conditions. However, they have also expanded the population of patients susceptible to opportunistic infections. Toxoplasma gondii (T. gondii), a globally prevalent parasite, has emerged as an underrecognized pathogen in this immunomodulated host population. Toxoplasmosis, in such patients, can occur either through reactivation of a chronic/latent/past infection or from an acute/primary infection and may be severe and even fatal. We present here the recommendations for such patients from the Remington Lab, the National Reference Center for Toxoplasmosis in the US. Screening for Toxoplasma infections is needed at baseline prior to starting targeted immunotherapy to identify seropositive patients who would benefit from prophylaxis or pre-emptive strategies and seronegative patients who would benefit from measures to prevent primary/acute infections. Prompt diagnosis of Toxoplasma disease (toxoplasmosis) with molecular tools (T. gondii PCR and/or agnostic metagenomics next-generation sequencing), and prompt initiation of anti-Toxoplasma therapy, can be lifesaving and prevent permanent neurocognitive sequelae and vision loss. The immunomodulatory effects of these therapies persist for several months after discontinuation, thereby extending the window of vulnerability. T. gondii-seropositive women are at increased risk of vertical transmission, even if targeted immunotherapy was discontinued several months before conception. We make a call for education, guidelines, prospective registries, targeted research, and addition of toxoplasmosis risk in the Warnings section of drug leaflets (and particularly so for T. gondii-seropositive women who intend to conceive after having been on targeted immunotherapies).

Pathogens doi: 10.3390/pathogens15070695

Authors: Xolani H. Makhoba

Polyamines such as putrescine, spermidine, and spermine play essential roles in most living organisms. They regulate fundamental processes, like cell proliferation, differentiation, growth, gene expression (DNA/RNA stability, transcription, and translation), and signal transduction. As important regulators, polyamines influence development, stress responses, and the progression of health and disease, including cancer and aging. These positively charged molecules have been extensively studied for decades. In humans, polyamines are often researched as potential therapeutic targets for diseases such as malaria and, more recently, COVID-19. Obligate parasites, such as Plasmodium falciparum, and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), rely on host cellular machinery for survival, replication, and growth. Notably, both hosts and pathogens need polyamines to sustain these processes. This review summarizes current advances in understanding the roles of polyamines in humans, viruses, and obligate parasites. It also explores strategies to prevent pathogens from hijacking host polyamine metabolism as a way toward developing novel therapeutic interventions.

Pathogens doi: 10.3390/pathogens15070694

Authors: José Mario Atondo-Pacheco Rosalino Flores-Rocha María de J. López-López Idalia Enríquez-Verdugo Daniel Eduardo Zatarain Jesús Daniel Solis-Carrasco Nohelia Castro-del Campo Sandra Berenice Medina-Rodríguez Soila Maribel Gaxiola-Camacho Nohemí Castro-del Campo

Zoonotic diseases represent an increasing public health concern worldwide, particularly in endemic regions of northwestern Mexico. This study aimed to evaluate household-level environmental and behavioral risk factors associated with Ehrlichia spp. infection in dogs living in areas with documented human rickettsiosis cases in Culiacán, Sinaloa, Mexico. A cross-sectional study was conducted, including 105 canine blood samples collected from urban and rural areas, with previous human rickettsiosis cases reported between 2021 and 2023. Microscopic examination and PCR amplification targeting the 16S rRNA gene of Ehrlichia spp. were performed. Morphological detection revealed a prevalence of 33.33%, while molecular analysis showed a prevalence of 43.83%. Risk factors significantly associated with infection included household waste, soil-type environments, free-roaming behavior identified through epidemiological analysis, and limited knowledge of tick-borne diseases among dog owners. These findings provide evidence of Ehrlichia spp. infection in dogs from areas with documented human rickettsiosis cases and highlight the value of dogs as indicators of household-level exposure to tick-borne pathogens in endemic communities.

Pathogens doi: 10.3390/pathogens15070693

Authors: Luca Dalle Carbonare Anna Vareschi Kevin Dervishi Michela Deiana Elena Locatelli Arianna Minoia Francesca Cristiana Piritore Alessandra Ruggiero Ilda Czobor Barbu Donato Zipeto Chiara Piubelli Maria Teresa Valenti

In this exploratory pilot study, quantitative analyses were performed on seven leather wheelchairs and the protective barrier was evaluated on three leather wheelchairs, while shotgun metagenomic sequencing (Illumina and Oxford Nanopore) was conducted on pooled samples obtained from seven leather and three fabric wheelchairs to characterize microbial DNA recovered from wheelchair surfaces under routine clinical conditions. Microbial DNA and biomass were detected on all sampled surfaces, with median DNA concentrations of approximately 0.015 ng/µL, median cell counts of approximately 4.8 × 105 cells/mL, and median OD600 values of approximately 0.038, although variability among wheelchairs was observed. NGS analysis revealed heterogeneous microbial communities composed mainly of taxa associated with human skin microbiota and environmental sources. Opportunistic taxa including Escherichia coli, Staphylococcus haemolyticus, Achromobacter xylosoxidans, and Clostridioides difficile DNA were detected. Differences in microbial composition were observed between the pooled fabric and leather samples, with fabric samples characterized by the dominance of specific taxa and leather samples exhibiting a more heterogeneous microbial profile. In addition, median DNA concentration, cell counts, and OD600 values were reduced by approximately 98–100% on the protective barrier compared with uncovered wheelchair surfaces, with statistically significant differences between conditions. Overall, these findings suggest that hospital wheelchairs may harbor measurable levels of microbial biomass and microbial DNA despite routine sanitation procedures. Lower contamination levels were observed on the protective barrier under the conditions tested. Due to the exploratory nature of the study, the small sample size, and the use of pooled samples for metagenomic analyses, these observations should be interpreted with caution and require confirmation in larger studies.

Pathogens doi: 10.3390/pathogens15070692

Authors: Xijie Ding Jianxing Li Guojun Chen Chaoyue Ji Weiguo Hu

Purpose: This study aimed to determine whether positive admission urine culture is associated with stone burden, renal involvement, pathogen distribution, and measured urinary biochemical profiles in men and women with calcium oxalate stones. Methods: We retrospectively analyzed adults who underwent percutaneous nephrolithotomy or ureterorenoscopy for upper urinary tract stones between 2016 and 2020. Calcium oxalate stones were defined as stones containing ≥50% calcium oxalate monohydrate and/or dihydrate by Fourier transform infrared spectroscopy. Patients were compared by sex and then stratified by admission urine culture status within each sex. Results: Among 1257 patients, 878 were men and 379 were women. Women had a higher culture-positive rate than men (59.6% vs. 28.2%, p < 0.001), despite lower 24-h urinary calcium, uric acid, sodium, potassium, phosphorus, and chloride. In men, culture positivity was associated with recurrent stones, renal stone location, larger maximum stone diameter, and lower eGFR, but not with measured 24-h urinary parameters. In women, culture positivity was associated with renal stone location, larger maximum stone diameter, lower eGFR, and modestly lower urinary calcium. Escherichia coli predominated among culture-positive women, whereas men showed a broader pathogen distribution. Conclusions: Positive admission urine culture was associated with greater stone burden and renal involvement in calcium oxalate stone disease, without a uniformly higher measured urinary biochemical profile. Culture status may provide clinically relevant phenotypic information alongside measured urinary biochemical assessment, although interpretation is limited by the absence of key CaOx-related urinary parameters such as oxalate, citrate, and supersaturation indices.

Pathogens doi: 10.3390/pathogens15070691

Authors: Juliana Aline Souza Lemos Barbara de Oliveira Baptista Carolina de Souza Faria Pereira Hugo Amorim dos Santos de Souza Jenifer Peixoto de Barros Rodrigo Medeiros Martorano Rodrigo Nunes Rodrigues-da-Silva Evelyn Kety Pratt Riccio Dave Richard Paulo Renato Rivas Totino Josué da Costa Lima-Junior Cláudio Tadeu Daniel-Ribeiro Lilian Rose Pratt-Riccio

Vacuolar Protein Sorting 29 (VPS29) is a highly conserved subunit of the retromer complex, which mediates retrograde transport from endosomes to the Golgi apparatus and plays a critical role in membrane trafficking, protein recycling, and organelle biogenesis. In Plasmodium falciparum, the retromer has been implicated in the formation of apical organelles essential for parasite invasion and replication. In this study, we investigated naturally acquired antibody responses to P. falciparum VPS29 (PfVPS29) and the genetic diversity of the vps29 gene in isolates from three malaria-endemic areas of the Brazilian Amazon. Naturally acquired responses to PfVPS29 were evaluated by ELISA in 533 individuals, and genetic diversity was assessed in 62 P. falciparum isolates. Only 17% of participants displayed IgG reactivity, whereas 73.5% showed IgM responses, indicating limited IgG acquisition but a predominant IgM profile associated with recent or ongoing exposure. IgG subclass analysis revealed a predominance of cytophilic IgG1 and IgG3 among responders. IgM responses were significantly boosted during P. falciparum infection. Sequence analysis revealed no polymorphisms among Brazilian isolates, and comparison with global datasets confirmed the high conservation of the PfVPS29 coding sequence. Together, these findings show that PfVPS29 is a highly conserved intracellular protein that elicits an atypical humoral response dominated by IgM, with limited class switching to IgG, like other conserved or repetitive malaria antigens. These results highlight PfVPS29 as an example of a conserved intracellular antigen that induces non-classical humoral responses in naturally exposed populations.

Pathogens doi: 10.3390/pathogens15070690

Authors: Sucharita Panigrahi Matrujyoti Pattnaik Rachita Pradhan Debaprasad Parai Shishirendu Ghosal Anoop Velayudhan Punit Prasad Adyasha Panda Debdutta Bhattacharya Sanghamitra Pati

Introduction: Integrated One Health-based surveillance of pathogens in wastewater suggests its potential for monitoring community health and preventing the emergence and spread of infectious diseases. Despite the growing popularity of Wastewater Surveillance (WWS) and its clinical utility, its uniformity remains poorly understood, especially concerning its clinical evidence. This review systematically synthesizes evidence on the role of wastewater surveillance in early pathogen detection and outbreak preparedness, with particular emphasis on its implications for One Health surveillance. Methods: We systematically searched PubMed, EMBASE, ProQuest and EBSCO CINAHL databases. Retrieved articles were screened by two reviewers, and conflicts were resolved by a third reviewer. Initially, 539 studies were retrieved as potentially eligible published articles, of which 16 articles fulfilled the inclusion criteria. Results: Most pathogens identified in the included studies were associated with respiratory and gastrointestinal infections. Studies found a positive link between the presence of pathogens in wastewater and clinical cases, depicting potential exposure and transmission within the communities. A season-specific upsurge was observed among the identified pathogens in circulation. In addition, the duration and frequency of sample collection in socio-vulnerable areas provide early warning of disease outbreaks. Few studies have explicitly operationalized a One Health framework, highlighting the need for integrated human, animal, and environmental surveillance systems in future wastewater surveillance programmes. Conclusion: The review emphasized wastewater surveillance as a promising complementary approach for the early detection and tracking of pathogens. Future research is needed to standardize surveillance approaches and strengthen One Health integration across human, animal and environmental health systems.

Pathogens doi: 10.3390/pathogens15070689

Authors: Bernal León Gabriel González Bradd Mendoza-Guido Consuelo Carrillo Luis L. Rodríguez Kathryn A. Hanley Nidia S. Trovao

Vesicular stomatitis virus (VSV) causes clinical disease in livestock that mimics Foot-and-Mouth Disease, necessitating its status as a reportable pathogen to the World Organization for Animal Health. Given the importance of accurate classification for epidemiological surveillance, this study aims to update VSV taxonomic organization using whole-genome sequence criteria to better monitor disease dynamics. Using phylogenetic analysis and the Species Demarcation Tool (SDT), we analyzed pairwise identity across publicly available sequences, constructing a rooted maximum likelihood tree to cluster strains based on robust genetic identity scores. The results demonstrate that nucleotide divergences exceeding 30% define distinct species within the Vesiculovirus genus. Within a species, divergences between 10% and 30% successfully delineate genotypes, while differences between 6% and 10% identify specific subtypes. These quantitative benchmarks provide a precise framework for viral classification, significantly enhancing genomic surveillance and the ability to track the evolution and transmission of VSV genotypes and subtypes across diverse geographic regions.

Pathogens doi: 10.3390/pathogens15070688

Authors: Sylwia Andrzejczuk Monika Wujec Łukasz Świątek Urszula Kosikowska

Dermatophytosis caused by Trichophyton spp. represents a growing public health concern, exacerbated by the increasing prevalence of difficult-to-treat infections and the emergence of resistance to standard antifungal agents, such as terbinafine. In this study, we evaluated the in vitro antifungal activity of a thiosemicarbazide derivative, 1-[(1-methyl-4-nitroimidazol-2-yl)carbonyl]-4-(3-methylphenyl)thiosemicarbazide, against a panel of anthropophilic and zoophilic Trichophyton spp. dermatophytes. Susceptibility testing was performed via the broth microdilution method to determine the minimum inhibitory concentrations (MICs) and minimum fungicidal concentrations (MFCs). The compound demonstrated antifungal efficacy against all the strains tested, with MIC values ranging from 31.25 to 125 µg/mL. Crucially, low MFC/MIC ratios (≤4) confirmed a fungicidal effect, which was further corroborated by microscopic analysis revealing severe hyphal damage in treated dermatophytes. The derivative exhibited consistent activity against both T. rubrum and T. mentagrophytes, including clinical isolates. To our knowledge, this is one of the first detailed evaluations of a nitroimidazole–thiosemicarbazide hybrid against Trichophyton spp. that combines quantitative MIC/MFC testing with morphological assessment. The tested thiosemicarbazide derivative noticeably decreased the viability of VERO and A375 cells, but even at the highest tested concentration, after 72 h of incubation, the cellular viability remained at 65%. These findings suggest that the investigated compound is a promising candidate for the development of new fungicidal drugs or disinfectants for the effective prophylaxis or management of superficial mycoses in human and veterinary medicine.

Pathogens doi: 10.3390/pathogens15070687

Authors: Li Li Xiaoqian Shi Mingyang Liu Tongzhen Xu Yinan Chen Ranjiaxi Wang Qiyue Zhang Dan Li

The intratumoral microbiota has emerged as a critical component of the tumor microenvironment (TME), with accumulating evidence indicating that its biological functions are influenced not only by microbial composition but also by their spatial organization within tumor tissues. This review summarizes the historical development and potential origins of intratumoral microbiota, and elaborates on the concept and biological significance of spatial heterogeneity. Based on recurrent spatial distribution patterns reported across different tumor types, we propose a conceptual framework comprising several putative spatial niches, including hypoxic/necrotic, immune-enriched, stromal-associated, invasive/metastatic, and intracellular niches. We further discuss the potential mechanisms contributing to the establishment and maintenance of spatial heterogeneity. The clinical significance of spatial microbial signatures is critically evaluated, alongside a comprehensive overview of spatial analytical methodologies, ranging from in situ hybridization and immunology-based approaches to emerging spatial omics and multi-omics integration strategies. Finally, we address key challenges and limitations, including contamination control, causal inference, barriers to clinical translation, and the underexplored spatial dimensions of the intratumoral mycobiome and virome. By synthesizing current knowledge and identifying critical gaps, this review aims to provide a conceptual and methodological framework for advancing spatially resolved investigations of intratumoral microbiota and facilitating their potential translational applications in precision oncology.

Pathogens doi: 10.3390/pathogens15070686

Authors: Verena Zerbato Stefano Guicciardi Roberto Baldan Chiara Fanelli Lisa Fusaro Alexandru Botan Nicola Benvenuto Giovanna Maria Nicolò Luigi Principe Dan Alexandru Toc Mauro Giuffrè Lory Saveria Crocé Luca Frulloni Abbas Yadegar Stefano Di Bella Alberto Enrico Maraolo

Purpose: Campylobacter spp. are a common cause of acute enteric infections. In immunocompromised or elderly patients, they can lead to extraintestinal infections, including bacteremia. The clinical significance of Campylobacter bacteremia is not fully understood. Methods: We conducted a systematic review and meta-analysis on Campylobacter spp. bacteremia, including studies published up to June 2024. Results: Twenty-five retrospective observational studies, published between 1978 and 2024, were included. The studies involved a total of 2480 patients, with a mean age range across studies from 1 to 70 years; 62.45% were male. The pooled prevalence of Campylobacter species was: C. jejuni 60% [95% CI 0.45–0.73], C. coli 8% [95% CI 0.04–0.13], C. fetus 7% [95% CI 0.03–0.15], and other species 9% [95% CI 0.04–0.16]. Mortality was the primary outcome in 22 studies, with a pooled case-fatality risk of 5% [95% CI 0.03–0.09]. Univariate meta-regression showed higher mortality associated with C. fetus (β = 3.217, [95% CI 0.632–5.802], p = 0.017), immunocompromised status (β = 2.749, [95% CI 0.316–5.184], p = 0.029), and chronic liver disease (β = 5.072, [95% CI 0.424–9.720], p = 0.034). Regarding complications, secondary localizations (e.g., endovascular infections) showed a pooled prevalence of 9% [95% CI: 0.04–0.18]; relapses, 3% [95% CI 0.02–0.04]; endocarditis, 2% [95% CI 0.01–0.03]; and persistent bacteremia, 1% [95% CI 0.001–0.27]. Conclusion: Campylobacter spp. bacteremia shows a considerable risk of mortality and complications.

Pathogens doi: 10.3390/pathogens15070685

Authors: Ziyat Abdel Zauresh Zhumadilova Raikhan Mussagalieva Aigul Abdirassilova Svetlana Issaeva Galina Kovaleva Bolatbek Baitursyn Beck Abdeliyev Temirkhan Sagidulin Nurbol Shaki Damira Shonshabayeva Alim Saduakas Tatyana Meka-Mechenko

Plague remains a significant natural focal zoonotic infection with continuing epidemiological relevance in the Republic of Kazakhstan. This study provides a comprehensive assessment of epizootological dynamics in natural plague foci during 2020–2025 through the integration of historical epidemiological data, phenotypic and molecular characterization of Yersinia pestis, and GIS-based spatial analysis. The study utilized long-term surveillance data (1920–2025), epidemiological records of human plague cases (1926–2003), phenotypic analysis of 1526 strains, and whole-genome sequencing of 75 representative isolates. Epizootological monitoring demonstrated high surveillance coverage and stable monitoring capacity, together with a marked increase in the application of molecular diagnostic methods. By 2025, both the number and isolation rate of Y. pestis strains increased substantially, while the epizootically active area expanded in 2024–2025, although the overall long-term trend in active area was not statistically significant. Despite these fluctuations, Y. pestis populations remained highly stable, with 94.9% phenotypically typical and 97.5% genotypically typical strains, and no evidence of antimicrobial resistance. Spatial autocorrelation analysis using Moran’s I revealed significant clustering of epizootics (Moran’s I = 0.1627; z = 4.39; p < 0.001), indicating non-random spatial distribution and localized zones of increased epizootic activity. No human plague cases have been recorded since 2003 during the period of sustained epidemiological surveillance and control measures. These findings highlight the potential utility of integrating spatial modeling and molecular surveillance into risk-oriented plague monitoring and control strategies.

Pathogens doi: 10.3390/pathogens15070684

Authors: Sawsan Mubarak Joud Jarrah Yara K. Edor Omar Khresat Hadeel AlGhawrie

Multidrug-resistant (MDR) pathogens present a significant threat to hematopoietic stem cell transplant (HSCT) recipients; despite their critical implications, regional data on their infection patterns remain scarce. This study aimed to characterize the incidence, pathogen and antimicrobial resistance distribution of clinically confirmed bacterial infections among HSCT recipients. A retrospective analysis was conducted at King Hussein Cancer Center, Jordan (2018–2022). MDR pathogens were defined per CDC criteria. During the study period, 1157 HSCT procedures were performed. A total of 327 patients developed clinically documented bacterial infections, yielding an overall cumulative incidence of 28.3%, with a higher burden in the pediatric cohort (34.7%), including exclusive identification of Klebsiella oxytoca in pediatrics (2.3%). Gram-negative bacteria dominated, with Escherichia coli (50.5%) and Klebsiella pneumoniae (22.0%) being most common. Extended-spectrum beta-lactamase (ESBL) production was the dominant resistance mechanism (71.3%), followed by carbapenem-resistant Enterobacteriaceae (CRE; 14.1%), methicillin-resistant Staphylococcus aureus (MRSA; 8.6%), and carbapenem-resistant Pseudomonas aeruginosa (CRPA; 7.0%). The urogenital (39.1%) and bloodstream (31.2%) were the most infected sites. Significant site-specific associations were noted for ESBL production, MDR-Acinetobacter baumannii (p < 0.001) and MRSA (p = 0.007). Temporal analysis revealed a convergent MDR peak in 2021. Our findings offer critical insights into MDR pathogen incidence in HSCT recipients in the Middle East, informing improved infection management and intensified antimicrobial stewardship in this high-risk population.

Pathogens doi: 10.3390/pathogens15070683

Authors: Miray Tonk-Rügen Ludwig E. Hoelzle Alejandro Cabezas-Cruz

The recent publication by Larska et al [...]

Pathogens doi: 10.3390/pathogens15070682

Authors: Kenichi W. Okamoto Luis F. Chaves Luke Bergmann Rodrick D. Wallace Robert G. Wallace

Public health policy foundationally impacts how pathogens spread, yet despite multiple pathogens of broader societal concern emerging, little research has examined how policy affects pathogen evolution. To evaluate this connection, we examine how varying public health approaches impact how viral immune susceptibility, including resistance to vaccines, evolves. Integrating evolutionary epidemiological modeling and critical geography, we compare how distinct public health responses early in the COVID-19 pandemic affected the potential evolution of immune evasion in SARS-CoV-2 across four territories: Costa Rica, Panama, Texas, and Uruguay. We use parameter estimates inferred from confirmed case and vaccination time series via stochastic ensemble Kalman filtering in each territory. Our analyses suggest viral immune resistance was most likely to emerge in Texas, which relied almost exclusively on vaccines for disease control. In contrast, regions with comparatively fewer health disparities that also rigorously applied interventions, such as shelter-in-place orders and household support, may have better prevented vaccine resistance from evolving. These comparative analyses highlight the key role policy choices play, potentially representing different governance goals for population health and wellbeing. We argue that such choices impact not only disease spread but also pathogen evolution along epidemiologically critical dimensions like viral immune susceptibility. Our study thus demonstrates how public health priorities drive social–evolutionary feedbacks.

Pathogens doi: 10.3390/pathogens15070681

Authors: Magdalena Stachnik Monika Normant-Saremba Anna Kycko

Hepatospora eriocheir is a microsporidian parasite of the Chinese mitten crab Eriocheir sinensis, an invasive decapod now widely distributed in European inland and coastal waters. Although the host is common across much of Europe, confirmed European records of H. eriocheir have remained scarce and, until now, have not included the Baltic Sea region. In this study, 15 adult E. sinensis collected from the Vistula Lagoon, southern Baltic Sea, were examined using gross pathological assessment, wet-mount microscopy, histopathology, PCR amplification, Sanger sequencing, and phylogenetic analyses of parasite SSU rRNA and host COI sequences. Hepatopancreatic alterations were observed in several individuals, ranging from pale discolouration and friability to loss of normal tissue organisation. Spores were detected in fresh squash preparations from affected tissue, and histology revealed epithelial disruption, intratubular spore accumulation, and necrotic changes consistent with progressive microsporidian infection. Molecular screening confirmed H. eriocheir in 60.0% of crabs, with positive cases occurring in both females and males. The parasite sequences formed a single, well-supported clade and were highly similar to previously reported H. eriocheir sequences from the United Kingdom and China. Host COI sequences represented three mitochondrial haplotypes, indicating that the infection occurred across more than one host mitochondrial haplotype background. These findings constitute the first record of H. eriocheir in E. sinensis from the Baltic Sea and support the hypothesis that infected crabs reaching the Vistula Lagoon are connected with the wider North Sea invasion system rather than an isolated Baltic lineage.

Pathogens doi: 10.3390/pathogens15070680

Authors: Shuo Wang Liwen Yin Jinhao Yang Changru Zhang Zhi Yao Xiaolei Pan

Pseudomonas aeruginosa can utilize abundant phosphatidylcholine (PC) and phosphatidylethanolamine (PE) within the host as energy and structural substrates. Fatty acids, choline, and ethanolamine liberated from PC and PE can each serve as the sole carbon source to support bacterial growth in vitro. Our previous work demonstrated that fatty acid metabolism is critical for acute pulmonary infection caused by P. aeruginosa. The pathogen senses host-derived fatty acids via the transcriptional regulator PvrA, which activates fatty acid utilization pathways and drives the production of virulence factors required for acute infection. In this study, we demonstrate that during acute pulmonary infection in mice, P. aeruginosa upregulates fatty acid catabolism while simultaneously repressing choline and ethanolamine uptake and metabolism pathways. Deletion of the transcriptional activators GbdR and EatR (which control choline and ethanolamine utilization respectively) enhances pulmonary bacterial colonization. We further identify fatty acids as environmental signals that trigger repression of choline and ethanolamine utilization programs. PvrA mediates this signaling cascade by directly binding to the promoters of gbdR and eatR and suppressing their transcription upon fatty acid exposure.

Pathogens doi: 10.3390/pathogens15070679

Authors: Maria Gavatha Emmanouil Angelos Rigopoulos Miranda Alexopoulou Vasileios Petrakis Nikoleta Babaka Olga Tsachouridou Dimitrios Pilalas Charis Chari Alexandra Vorria Evaggelia Bogosian Petros Ioannou Sofia Ioannou Efstratios Patsatzis Maria N. Gamaletsou Andreas Rafail Tzatzimos Periklis Panagopoulos Symeon Metallidis Dimitrios Papazoglou Konstantinos Tosios Karolina Akinosoglou

Oral manifestations are common in people living with HIV (PLWH) and may affect oral health-related quality of life (OHRQoL), while data from Greece remain limited. This multicenter prospective cohort study evaluated oral health status and OHRQoL among PLWH and explored associations with antiretroviral therapy (ART) and clinical factors. Overall, 370 PLWH from seven referral centers were included. Participants underwent oral examination, with oral hygiene assessed using the Simplified Oral Hygiene Index (OHI-S), and completed the Oral Health Impact Profile-14 (OHIP-14). Statistical analyses were performed using IBM SPSS Statistics v29.0, while multinomial and binary logistic regression identified predictors of oral hygiene status and OHRQoL, respectively. Most participants were male (76.5%), had CD4 counts ≥ 200 cells/μL (95.4%), and were receiving ART (98.6%). Annual dental check-ups, daily tooth brushing, mouthwash use, and dental floss use were reported by 54.1%, 69.5%, 31.9%, and 23.8%, respectively. The median OHI-S score was 2.0 (IQR:1.5–2.7), with 16.9% having poor OHI-S; the median OHIP-14 score was 11 (IQR: 7–15), with 64.4% reporting poor OHRQoL. Male sex was associated with lower odds of poor OHRQoL (OR = 0.377; p = 0.006), whereas ART regimen independently predicted poor OHRQoL. These findings support patient-centered oral healthcare within HIV care.

Pathogens doi: 10.3390/pathogens15070678

Authors: Katrina Warnes Daniel S. Mills Andrew S. Cooke Stefan H. Millson Simon R. Clegg

Vaccination is one of the most important measures for infectious disease control. Recently, media-generated concern about vaccine-associated adverse effects has produced a rise in both human and animal “anti-vaccination” movements. This study aimed to understand factors involved in dog owner vaccination decisions and explore whether there has been an increase in titer testing. An online survey targeting dog owners received a total of 2585 responses, which showed 79% of respondents had their dogs vaccinated in the last 12 months. A few owners had never vaccinated their dogs, and 13% of owners used titer testing prior to booster vaccinations. The factors with the strongest positive predictors for vaccination were requirements by third party services (e.g., kennels) and, for a negative response, lack of time. For respondents that had not vaccinated, the factors with the strongest predictive powers to determine if they titer test were education/working in the veterinary industry for a positive response and not having heard of negative side effects after vaccinating for a negative response. Overall, no evidence was found that a rise in anti-vaccination attitudes was pervasive in dog owners; however, the study shows that the veterinary profession has work to do to ensure herd immunity is maintained within dog populations.

Pathogens doi: 10.3390/pathogens15070676

Authors: Murimisi Mukansi Helen C. Steel Theresa M. Rossouw Ismail Kalla Colin Menezes Martin Nieuwoudt Ronald Anderson Charles Feldman

No previous sub-Saharan studies have compared patients with community-acquired pneumonia (CAP) and COVID-19 pneumonia, the focus of this study. Consecutive adult patients hospitalized with CAP (n = 59) or COVID-19 pneumonia (n = 74) were compared regarding multiple characteristics, including cardiac biomarkers. In multivariable logistic regression analysis, differences were noted among various clinical features. Troponin I concentrations (p = 0.00028) and the Troponin I/NT-pro BNP ratio (p = 0.00048) were significantly higher in COVID-19 compared with CAP. After adjustment for age, these differences remained significant (troponin I p = 0.0019; ratio p = 0.00054), while BNP concentrations were now higher in CAP (p = 0.009). PCA demonstrated that BNP and NT-pro BNP contributed most strongly to the dominant cardiac biomarker signature, suggesting shared cardiopulmonary stress across both diseases. Exploratory subgroup analyses suggested higher troponin I levels among people living with HIV and COVID-19, although interaction modelling did not demonstrate significant effect modification by HIV status. Both CAP and COVID-19 pneumonia were associated with evidence of cardiac stress; however, COVID-19 demonstrated a relatively stronger myocardial injury signature characterized by higher troponin I concentrations and an increased Troponin I/NT-pro BNP ratio while CAP had evidence of greater hemodynamic cardiac strain, as evidenced by the higher levels of BNP. The findings suggest that the mechanisms of cardiac involvement may differ between viral and bacterial respiratory infections.

Pathogens doi: 10.3390/pathogens15070677

Authors: Gregory D. Hawley Ambika Agrawal Maryam Alhashmi Milica Novakovic Andrea K. Boggild

Leptospirosis is an emerging zoonotic infectious disease in the adventure traveler with recreational freshwater exposure. We describe three cases of probable leptospirosis acquired during whitewater rafting trips in rural Ecuador. Rafting took place following periods of heavy rainfall and flooding of rivers, which are well documented risk factors for leptospirosis. Two of our patients presented to our rapid assessment clinic with acute febrile illness after travel to a region with a confirmed leptospirosis outbreak, while the third patient presented four weeks after a convalescent episode of acute febrile jaundice during a dramatic rise in reported leptospirosis cases in Ecuador. Delayed turnaround times for confirmatory microbiologic diagnostics in leptospirosis challenge its management and necessitate provisional clinical diagnosis and empiric antimicrobials based on compatible clinical symptoms, biochemical abnormalities, exposure history, and known outbreak epidemiology. We herein situate the presentations of our patients within the broader literature by reviewing the clinical, diagnostic, therapeutic, and prognostic considerations when caring for leptospirosis patients. Leptospirosis should be considered in the differential diagnosis of all patients presenting with acute febrile illness following adventure travel with freshwater exposure, and empiric treatment should be considered given the absence of readily available confirmatory microbiologic testing.

Pathogens doi: 10.3390/pathogens15070675

Authors: Humberto Pires Sónia Saraiva Manuela Matos Ana Patrícia Lopes Maria da Conceição Fontes Cristina Pintado Luís Figueira Sérgio Santos-Silva Ana Cristina Matos Ana Cláudia Coelho Luís Cardoso João Rodrigo Mesquita

Tularemia is a zoonotic disease caused by the highly contagious bacterium Francisella tularensis, with rabbits, hares, and rodents considered as primary reservoirs. Clinical manifestations in humans can range from mild to life-threatening, depending on the mode of exposure (ingestion, inhalation, skin contact, injection, or tick bite), with high fever being a common feature. The primary aim of the present research was to evaluate the circulation of F. tularensis and the potential infection among wild boar (Sus scrofa) and red deer (Cervus elaphus) from the central region of Portugal. A total of 368 samples (including serum and organ samples) were collected from 184 wild boar and 184 red deer. For serological analysis, two commercially available enzyme-linked immunosorbent assay (ELISA) kits were used to detect immunoglobulin M (IgM) and G (IgG) antibodies to F. tularensis. Antibodies to F. tularensis (IgG and IgM) were detected in one adult male wild boar, corresponding to a seroprevalence of 0.54% (1/184, 95% confidence interval [95% CI]: 0.01–3.0%). No antibodies to F. tularensis were detected in red deer. Molecular detection by PCR was negative in the seropositive animal, in which submandibular lymph node, liver, and spleen samples were analysed targeting the 16S rRNA gene. These findings indicate exposure of wild boar to F. tularensis in central Portugal, suggesting a sporadic presence of the pathogen. Although no evidence of active infection was detected in the analysed tissues, the presence of seropositive individuals highlights the need for further investigation. Despite the very low seroprevalence observed, the zoonotic potential of F. tularensis supports the importance of continued surveillance within a One Health perspective.

Pathogens doi: 10.3390/pathogens15070674

Authors: Elena Teona Cosovanu Silvia Ionescu Eric Oliviu Cosovanu Costin Damian Bogdan Aurelian Stana Ecaterina Iftime Antoneta Dacia Petroaie Tiberiu Lunguleac Ileana Katerina Ioniuc Elena Adorata Coman Cristina Daniela Dimitriu Demetra Gabriela Socolov Luminita Smaranda Iancu Irina Draga Caruntu Ramona Gabriela Ursu

Background: Early-onset neonatal listeriosis is a rare, life-threatening infection of vertical origin caused by Listeria monocytogenes. First-line therapy is intravenous ampicillin combined with an aminoglycoside; acquired β-lactam resistance is exceptionally uncommon. Case Presentation: A 34-week preterm female neonate (birth weight 1990 g, appropriate for gestational age) was born to a febrile primigravida with fetid greenish amniotic fluid at a regional secondary maternity and transferred at 30 h of life to our tertiary NICU with respiratory failure requiring mechanical ventilation. L. monocytogenes was recovered from blood, gastric aspirate, pharyngeal exudate, ocular secretion, and skin swab. Gradient strip susceptibility testing reported ampicillin and trimethoprim–sulfamethoxazole non-susceptibility, although confirmatory broth microdilution was unavailable. Broad-spectrum empirical therapy was revised on Day 5 to include ampicillin–sulbactam, with piperacillin–tazobactam and gentamicin continued. A follow-up blood culture on Day 9 remained sterile through 7 days. The hospital course was complicated by thrombocytopenia, transiently elevated aminotransferases, and a Grade I subependymal hemorrhage; tertiary NICU length of stay was 25 days. Conclusions: Recovery under a multi-agent regimen precludes attribution of effect to any single component. Discordant gradient strip susceptibility results in L. monocytogenes should be confirmed by broth microdilution before any therapeutic change; survivors of severe early-onset listeriosis require structured multidisciplinary follow-up.

Pathogens doi: 10.3390/pathogens15070673

Authors: Reuven Yosef

Domestic cats (Felis catus) are ubiquitous companion animals that provide substantial psychological and social benefits to children and adults alike, but they also serve as reservoirs and vectors for a wide range of zoonotic pathogens. Close physical contact between cats and children, frequent use of shared environments such as homes, playgrounds, and sandboxes, and still-developing hygiene behaviours increase opportunities for exposure to protozoa, helminths, bacteria, fungi, and ectoparasite-borne agents. This review synthesizes current evidence on key feline-associated zoonoses of pediatric concern—including Toxoplasma gondii, Toxocara cati, Ancylostoma spp., Dipylidium caninum, Bartonella henselae, Salmonella enterica, Campylobacter jejuni, Pasteurella multocida, Microsporum canis, flea-borne Rickettsia species, and rabies—with emphasis on transmission routes, clinical manifestations, and risk modifiers in children, pregnant women, and immunocompromised individuals. Within a One Health framework, we also summarize global publication trends on feline zoonoses, discuss how urban cat ecology and management (including free-ranging cats in child-frequented environments) may shape pediatric risk, and outline practical prevention strategies centred on hygiene, veterinary care, and targeted education for caregivers and children.

Pathogens doi: 10.3390/pathogens15070672

Authors: D. P. Grosa D. Zavadska Z. Freimane L. Karele G. Karelis

Background: Tick-borne encephalitis (TBE) is an endemic neuroinfectious disease prevalent in parts of Europe and is often associated with persistent neurological and cognitive sequelae. The aim of this study was to evaluate the long-term outcomes and predictors of post-encephalitic sequelae in adult patients with TBE in Latvia. Methods: A retrospective cohort with prospective follow-up was used that included 105 adult patients hospitalized with laboratory-confirmed TBE between 2018 and 2024. The patients’ clinical and demographic data were extracted from medical records, and reassessments were performed ≥6 months after discharge using structured clinical and neurological evaluations for neurocognitive, subjective, and neurological sequelae. Disease severity was classified using the Mickienė and Bogovič criteria, and sequelae severity was defined according to the Bohr criteria for post-encephalitic syndrome (PES). Results: Sequelae were observed in 52/105 (49.5%) patients and were more frequent in meningoencephalitis than in meningitis cases (18/25 [72.0%] vs. 33/77 [42.9%]). The most common persistent symptoms were impaired concentration (33/52 [63.5%]), fatigue (29/52 [55.8%]), and sleep disturbances (21/52 [40.4%]). Neurological sequelae included tremor (23/52 [44.2%]), vertigo (11/52 [21.2%]), and hearing impairment (5/52 [9.8%]). According to the Bohr criteria, most of the patients had mild sequelae (42/52 [80.8%]), while 10/52 [19.2%] had moderate sequelae; no severe cases were observed. In the multivariable analysis, increasing age was independently associated with greater sequelae severity (OR = 1.045 per year; 95% CI, 1.015–1.073; p = 0.003). Sex, comorbidities, biphasic disease, and length of hospital stay were not significant predictors. Acute neurological manifestations, particularly paresis (p = 0.002) and tremor (p = 0.019), were associated with worse outcomes. Although the disease severity scores correlated with sequelae in unadjusted analyses, neither the Mickienė nor the Bogovič classification independently predicted outcomes after adjustment. Conclusions: Nearly half of the hospitalized patients with TBE included in this study developed long-term sequelae, which were predominantly neurocognitive and mild in severity. Age was the primary independent predictor of worse outcomes, while acute neurological deficits such as paresis and tremor also indicated increased risk. These findings highlight the substantial burden of post-encephalitic syndrome and the need for structured long-term follow-up in TBE survivors.

Pathogens doi: 10.3390/pathogens15070671

Authors: Antonios Synapalos Anastasia Diakou Stefanos Sgardelis

Brown bear populations in Greece face multiple threats, and parasitic infections may pose an additional risk to these vulnerable animals. This study represents the first comprehensive assessment of endoparasite occurrence, prevalence, and seasonality in brown bears in Greece, in relation to geographical location and the animal’s different physiological phases. A total of 918 faecal samples were collected over a three-year period from regions with brown bear presence in Greece. For each sample, the date of collection and the coordinates of the site were recorded. Samples were examined using sedimentation, flotation, and McMaster techniques, while the Baermann method was additionally applied to a subset of 195 samples. Spatial and temporal patterns in parasite occurrence and diversity were analysed using generalised additive models (GAMs). Ten parasitic taxa were identified, with Baylisascaris transfuga being the most prevalent (39.8%), followed by Crenosoma spp. (26%), Uncinaria spp. (18.09%), and Dicrocoelium dendriticum (14.38%). Less prevalent taxa included Eucoleus aerophilus, Sarcocystis spp., Toxascaris leonina, Eimeria spp., Linguatula serrata, and Taeniidae. Μixed infections, involving two or more parasites, were detected in 22% of the samples. The prevalence of B. transfuga was higher in late autumn, with high-risk infection areas identified in both late summer and autumn. In contrast, Uncinaria spp. and D. dendriticum showed no seasonal variation, while D. dendriticum exhibited spatial clustering patterns similar to B. transfuga but without clear seasonal trends. These findings highlight the widespread occurrence and complexity of parasitic infections in Greek brown bears. Continued long-term monitoring is essential to improve understanding of transmission dynamics and the ecological processes shaping parasite distribution in this animal species.

Pathogens doi: 10.3390/pathogens15070670

Authors: Peter Collignon John J. Beggs Jan M. Bell Denise Daley Elizabeth Roughead on behalf of the Australian Group on Antimicrobial Resistance on behalf of the Australian Group on Antimicrobial Resistance

Background: Antimicrobial resistance (AMR) is widely considered to be driven by antimicrobial consumption through within-host selection. However, whether this mechanism adequately explains population-level patterns of resistance in invasive infections remains uncertain. If antimicrobial use is the dominant determinant, resistance should be highest in demographic groups with the greatest exposure. Methods: We conducted a retrospective analysis of 44,792 community-onset Escherichia coli bloodstream infection episodes identified through national Australian surveillance data (2013–2024). Resistance prevalence across individual antimicrobials and composite multidrug resistance panels was analysed by age and sex. These data were compared with community antimicrobial dispensing derived from the Pharmaceutical Benefits Scheme. Mean resistance was modelled as a function of age and sex. Results: Antimicrobial use was substantially higher in females than males (~23% overall) and increased markedly with age, with individuals aged ≥80 years receiving approximately three times more antimicrobials than those aged 25–30 years. In contrast, resistance was consistently lower in females across most antimicrobials and composite measures. Resistance demonstrated an inverted U-shaped age distribution, peaking at 30–40 years before declining in older age groups. From early adulthood to older age, antimicrobial dispensing increased threefold, whereas mean resistance declined by approximately 20%. These patterns were consistent across antimicrobial classes, years, and jurisdictions. Conclusions: These findings show that demographic patterns of antimicrobial resistance in community-onset E. coli bloodstream infections are not well explained by a simple population-level consumption model. These findings should be interpreted as important hypothesis-generating insights. Although antimicrobial exposure remains important for individual-level selection, the observed discordance between prescribing and resistance suggests that other factors, including differences in transmission pathways, healthcare contact, disease prevalence, community sanitation and socioeconomic circumstances may also significantly shape resistance patterns.

Pathogens doi: 10.3390/pathogens15070669

Authors: Raikhan Mussagalieva Ziyat Abdel Zauresh Zhumadilova Aigul Abdirassilova Svetlana Issaeva Bolatbek Baitursyn Nurbol Shaki Beck Abdeliyev Dinmukhammed Otebay Tatyana Meka-Mechenko

Camels are increasingly recognized as an important epizootological and epidemiological link in natural plague foci, contributing to the transmission of Yersinia pestis from wildlife to humans. In the Republic of Kazakhstan, where natural plague foci occupy up to 40% of the territory, the rapid growth of camel populations may significantly enhance epidemiological risks. The aim of this study was to perform a comprehensive assessment of the role of camels in the epizootology and epidemiology of plague based on retrospective data (1907–2003) and contemporary monitoring (2000–2025), including spatial analysis and risk zoning. A total of 64 cases of camel plague and 43 epizootic foci were identified during the historical period. More than 400 human cases, including fatal outcomes, associated with infected camels were documented, with direct contact during slaughter and meat processing accounting for 94.7% of infections. Spatial analysis and epidemiological zoning revealed a heterogeneous risk distribution, with western and southern regions representing the highest-risk areas. Serological investigation (n = 2726) showed 75.6% seropositivity, likely reflecting substantial population immunity largely associated with vaccination. Despite increasing camel population size (from 227.7 to 304.0 thousand heads in 2020–2025), vaccination coverage varied between 32.0% and 51.0%, reflecting risk-based preventive strategies. The absence of recent camel plague cases supports the effectiveness of integrated control measures, including vaccination, surveillance, and the establishment of protective zones. These findings suggest that camels remain an important component of plague transmission systems and should be systematically integrated into surveillance programs within a One Health framework.

Pathogens doi: 10.3390/pathogens15070668

Authors: Alexander O. Frolov Anna I. Solovyeva Marina N. Malysheva Maria E. Belokon Grigory N. Machakhtyrov Varvara A. Machakhtyrova Anatoly A. Bondarev Maria S. Maximova Anna I. Ganyukova

The subgenus Megatrypanum Hoare, 1964, with the type species Trypanosoma theileri Laveran, 1902, comprises stercorarian trypanosomes of mammals. A substantial portion of this subgenus consists of T. theileri-like trypanosomes parasitizing cervids and bovids worldwide. Similar to most other members of the genus Trypanosoma that lack obvious economic importance, the biology of T. theileri-like trypanosomes remains poorly understood. In particular, fundamental aspects such as their host specificity, host–parasite interactions, and the morphology of developmental stages have been studied only to a limited extent. In this work, we provide a detailed description of the development and cellular organization of T. theileri-like trypanosomes in the horsefly Hybomitra montana using transmission electron microscopy (TEM). We show for the first time that T. theileri-like trypanosomes possess a well-developed cytostome–cytopharyngeal complex, morphologically similar to those in other stercorarian trypanosomes. This complex is present in the studied trypanosomes at the epimastigote stage and degrades during metacyclogenesis. In the host ileum, epimastigotes and trypomastigotes at different stages of metacyclogenesis are embedded in a fibrillar matrix that isolates them from the gut lumen. This promotes their accumulation in the vector, thereby increasing the efficiency of future infection of the vertebrate host, which occurs via contamination of the oral mucosa.

Pathogens doi: 10.3390/pathogens15070667

Authors: Marianna Imparato Annalisa Buonanno Angela Maione Monica Matuozzo Chiara D’Ambrosio Andrea Scaloni Marco Guida Emilia Galdiero Elisabetta de Alteriis

This study describes extracellular vesicles (EVs) isolated from the culture supernatant of a Kluyveromyces marxianus strain deriving from an artisanal sourdough. Previous work had clearly shown the probiotic properties of the yeast isolate and its antagonistic activities against clinical fluconazole-resistant Candida albicans strains. Characterization of the isolated EVs by nanotracking particle analysis showed they had a mean diameter of 157.7 nm. Proteomic characterization of the purified EVs identified a complex array of 100 proteins. Both C. albicans planktonic growth and biofilm formation were inhibited by K. marxianus EVs, as well as adhesion and invasion of Candida cells in the vaginal epithelial A-431 cells. In the same cell model, K. marxianus EVs exerted an immunomodulatory effect affecting the secretion of pro-inflammatory and anti-inflammatory cytokines. Further, the expression of C. albicans SAP2 and SAP6 genes, coding for two aspartyl proteases involved in the invasion and damage of the epithelial mucosa, was affected by the presence of the yeast EVs. Overall, the results of this study show that K. marxianus EVs retain, at least in part, the beneficial features of the live microorganism, representing a postbiotic cell-free alternative preparation potentially useful for the management of C. albicans vaginal infections.