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Keywords = fungal identification

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24 pages, 3604 KB  
Article
From Species Identification to Empirical Therapy: A Machine Learning and Rule-Based Decision Support Framework for Antifungal Resistance Prediction in ICU Candida Infections
by Madalina (Preda) Solomon, Beatrice Mahler, Lia-Mara Ditu, Oana Popescu, Corina-Aurelia Zugravu and Loredana Sabina Cornelia Manolescu
Med. Sci. 2026, 14(2), 319; https://doi.org/10.3390/medsci14020319 (registering DOI) - 15 Jun 2026
Abstract
Objectives: When a Candida species is identified in an ICU patient, susceptibility results are typically available in 24–72 h. In this study, we built a machine learning model using four variables available at identification to estimate resistance probability in real time. Methods [...] Read more.
Objectives: When a Candida species is identified in an ICU patient, susceptibility results are typically available in 24–72 h. In this study, we built a machine learning model using four variables available at identification to estimate resistance probability in real time. Methods: We analysed 747 fungal isolates from 725 ICU patients (January 2021–March 2026). We trained and compared a Random Forest and a Logistic Regression model, evaluating both with temporal cross-validation, permutation feature importance, three-category (S/I/R) prediction, and calibration analysis. Results: Multidrug resistance doubled from 24.5% (2021) to 51.1% (2025), and Candida auris grew eight-fold in three years. Random Forest reached AUC 0.885 on the held-out test set and 0.848 on prospective 2024–2025 data (Brier score 0.093). Species identity and drug choice together explained 87% of predictive signal. Local C. albicans fluconazole resistance (~16%) far exceeded the ECMM European figure of 0%, and C. krusei was four times more prevalent than the continental average. Conclusions: A four-variable model may provide calibrated resistance estimates during the critical gap before susceptibility results return, though performance reflects predominantly deterministic species–drug patterns rather than complex learned biology. Overall performance was comparable to a rule-based lookup table, confirming that the majority of predictive signal derives from established species–drug susceptibility patterns. Meaningful added value is limited to temporal trend tracking and improved prediction where resistance is acquired rather than intrinsic (C. albicans, C. tropicalis hard-subset AUC 0.929 vs. rule-based 0.899). The model complements a local antifungal testing; it does not replace one. Full article
(This article belongs to the Section Immunology and Infectious Diseases)
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24 pages, 695 KB  
Review
Recent Outbreaks, Resistance Trends, and Control Measures in Candida auris and Candida glabrata Infections
by Sepinoud Raeisi, Priya Madhavan and Diajeng Sekar Adisuri
J. Fungi 2026, 12(6), 436; https://doi.org/10.3390/jof12060436 (registering DOI) - 15 Jun 2026
Abstract
The global rise in multidrug-resistant (MDR) fungal pathogens has positioned Candida auris and Candida glabrata as major threats to public health. In recent years, these pathogens have increasingly been reported beyond traditional hospital settings, including neonatal intensive care units, long-term care facilities, oncology [...] Read more.
The global rise in multidrug-resistant (MDR) fungal pathogens has positioned Candida auris and Candida glabrata as major threats to public health. In recent years, these pathogens have increasingly been reported beyond traditional hospital settings, including neonatal intensive care units, long-term care facilities, oncology wards, and post-pandemic critical care environments. International surveillance bodies, including the Centers for Disease Control and Prevention (CDC), European Centre for Disease Prevention and Control (ECDC), World Health Organization (WHO), and regional monitoring networks, have documented escalating antifungal resistance, complex outbreak dynamics, and persistent gaps in infection control implementation. C. auris has emerged as a major etiological agent of healthcare-associated outbreaks, particularly in intensive care and neonatal units. Surveillance data indicate that a high proportion of C. auris isolates exhibit resistance to azoles, often exceeding 80% in some regions, while echinocandin resistance remains variable. Resistance patterns have evolved from predominantly azole resistance to broader multidrug-resistant phenotypes, including treatment-emergent echinocandin resistance. Six genetically distinct clades (I–VI) have been identified, with Clades I, III, and IV associated with large-scale outbreaks, whereas available data suggests that Clades II, V, and VI are more geographically restricted, although evidence for the recently described clades remains limited. C. glabrata is increasingly recognized as a major cause of invasive candidiasis, with rising resistance reported across multiple regions. While reduced azole susceptibility was historically predominant, emerging evidence highlights rising dual azole–echinocandin resistance, adaptive microevolution during antifungal therapy, and biofilm-associated tolerance mechanisms. Despite these advances, significant gaps persist in global resistance surveillance and in the mechanistic understanding of virulence and antifungal adaptation. Current mitigation strategies include antifungal stewardship programs, expanded resistance testing, and strengthened surveillance systems. Advances in rapid diagnostic technologies such as matrix-assisted laser desorption ionization–time of flight (MALDI-TOF) mass spectrometry, polymerase chain reaction (PCR)-based assays, and genomic surveillance have improved pathogen identification and outbreak detection, although accessibility remains limited in resource-constrained settings. This review examines emerging epidemiological, genomic, and antifungal resistance trends in C. auris and C. glabrata and highlights key priorities for improving diagnosis, surveillance, stewardship, and management of multidrug-resistant Candida infections. Full article
(This article belongs to the Special Issue Multidrug-Resistant Fungi, 2nd Edition)
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15 pages, 664 KB  
Article
Escalating Antifungal Resistance Among Candida Species in Reproductive-Age Women in Vietnam: Implications for Women’s Health and Healthcare Systems
by Bac V. G. Nguyen, Tu Thien Nhat Nguyen, Bang Chau Ngoc Tu, Hung Van Cao, Bich Ngoc Thi Nguyen, Thanh Tri Vu, Gia-Phong Vu, Hoai Thu Le and Phuoc Vinh Nguyen
Pathogens 2026, 15(6), 625; https://doi.org/10.3390/pathogens15060625 - 11 Jun 2026
Viewed by 96
Abstract
Vulvovaginal candidiasis (VVC) is a common fungal infection among reproductive-age women and is increasingly challenged by the emergence of non-albicans Candida species and reduced azole susceptibility. This prospective cross-sectional study investigated 235 symptomatic reproductive-age women attending two healthcare facilities in Ho Chi [...] Read more.
Vulvovaginal candidiasis (VVC) is a common fungal infection among reproductive-age women and is increasingly challenged by the emergence of non-albicans Candida species and reduced azole susceptibility. This prospective cross-sectional study investigated 235 symptomatic reproductive-age women attending two healthcare facilities in Ho Chi Minh City, Vietnam, to determine VVC prevalence, Candida species distribution, pregnancy-associated patterns, antifungal susceptibility, and diagnostic performance. Vaginal swabs were cultured on Sabouraud Dextrose Agar and CHROMagar™ Candida, while species identification was confirmed by PCR-RFLP targeting the ITS region. Susceptibility to fluconazole and clotrimazole was assessed using the disk diffusion method. Candida spp. was detected in 55.7% of participants. C. albicans accounted for 50.3% of isolates, whereas non-albicans Candida species represented 49.7%, indicating a substantial species shift. VVC was more frequent among pregnant women, particularly in the third trimester. Most C. albicans, C. tropicalis, and C. parapsilosis isolates remained susceptible to azoles; however, C. glabrata showed markedly reduced susceptibility to fluconazole and clotrimazole. CHROMagar™ Candida reliably identified C. albicans but misclassified several non-albicans Candida isolates compared with PCR-RFLP. These findings highlight the need for routine species-level diagnosis, antifungal susceptibility testing, and strengthened VVC surveillance in reproductive and antenatal healthcare settings in Vietnam. Full article
17 pages, 2466 KB  
Article
Rapid Culture-Independent Detection of Fish Pathogens Using Oxford Nanopore Technologies: Case-Based Insights Across Multiple Species and Tissues
by Konrad Wojnarowski, Paulina Cholewińska, Dongqing Zhao, Yoshikazu Hasegawa, Daniela Denk and Dušan Palić
Pathogens 2026, 15(6), 622; https://doi.org/10.3390/pathogens15060622 - 10 Jun 2026
Viewed by 175
Abstract
Rapid and accurate diagnosis of infectious diseases in aquaculture is essential for preventing major economic and ecological losses. Traditional culture-based methods focus on isolation of individual pathogens, and often are burdened with extended processing times, particularly during investigations of polymicrobial infections. Application of [...] Read more.
Rapid and accurate diagnosis of infectious diseases in aquaculture is essential for preventing major economic and ecological losses. Traditional culture-based methods focus on isolation of individual pathogens, and often are burdened with extended processing times, particularly during investigations of polymicrobial infections. Application of Oxford Nanopore Technologies (ONT) sequencing offers a rapid, culture-independent workflow for the identification of bacterial and fungal pathogens directly from fish tissues. Swab and organ samples from four cases (1: Salmo spp.; 2: Cyprinus carpio; 3: Salvelinus fontinalis; 4: Heniochus acuminatus) were analyzed using ONT long-read sequencing for metagenomic screening and bioinformatic classification. The results revealed case-, species-, and tissue-specific microbial profiles, with external tissues showing higher microbial diversity and internal organs enriched in pathogenic taxa. Dominant pathogens included Streptococcus iniae, Aeromonas hydrophila, Pseudomonas spp., and Saprolegnia parasitica, alongside opportunistic zoonotic bacteria such as Escherichia coli and Acinetobacter baumannii. We demonstrate the potential for diagnostic application of ONT sequencing in investigations and detection of multi-pathogen infections, including assessments of microbial community structure changes during disease outbreaks in aquatic species. The presented workflow enables rapid, cost-effective, and comprehensive pathogen profiling, supporting early disease surveillance and improved management in aquatic veterinary practice. Full article
(This article belongs to the Special Issue Recent Advances in the Diagnosis of Fish Pathogens)
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20 pages, 23407 KB  
Article
Comprehensive Analysis of IDD Transcription Factors and Their Expression Profiling Under Pathogen Stress in Wheat
by Yanzhen Wang, Shikai Lyu, Yanqi Wang, Jialu Li, Xia Liu and Menglin Lei
Biology 2026, 15(12), 904; https://doi.org/10.3390/biology15120904 - 9 Jun 2026
Viewed by 210
Abstract
INDETERMINATE DOMAIN (IDD) transcription factors are plant-specific regulators essential for plant development and stress adaptation. As a globally important staple crop, common wheat (Triticum aestivum L.) is frequently threatened by fungal diseases such as powdery mildew and stripe rust. To date, however, [...] Read more.
INDETERMINATE DOMAIN (IDD) transcription factors are plant-specific regulators essential for plant development and stress adaptation. As a globally important staple crop, common wheat (Triticum aestivum L.) is frequently threatened by fungal diseases such as powdery mildew and stripe rust. To date, however, the IDD gene family in wheat has not been systematically characterized, and its roles in biotic stress responses remain unclear. In this study, we performed genome-wide identification and a comprehensive analysis of the TaIDD gene family. A total of 41 TaIDD genes were identified, which were unevenly distributed across 15 chromosomes and divided into four phylogenetic groups. Synteny and selective pressure analyses demonstrated that segmental duplication was the main driver of family expansion and that TaIDD genes underwent strong purifying selection during evolution. Cis-acting element analysis revealed abundant hormone- and stress-related elements in their promoter regions. Transcriptome and RT-qPCR analyses indicated that TaIDD genes exhibited distinct expression patterns under abiotic and biotic stress. Notably, TaIDD13, TaIDD19, TaIDD27, TaIDD37, TaIDD39, and TaIDD41 were significantly induced by multiple fungal pathogens, suggesting their potential involvement in stress-responsive pathways that may be related to disease resistance. Subcellular localization analysis further confirmed that TaIDD39 was exclusively localized in the nucleus, consistent with its function as a transcriptional regulator. Our findings provide insights into the evolutionary characteristics and stress-response mechanisms of TaIDD genes and highlight TaIDD39 and other potential candidates that may serve as valuable resources for wheat molecular breeding to enhance broad-spectrum disease resistance and stress tolerance. Full article
(This article belongs to the Section Plant Science)
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14 pages, 2039 KB  
Article
Isolation and Characterization of Yeast-like and Filamentous Fungi from Lutzomyia longipalpis (Lutz & Neiva, 1912)
by Lidiane da Silva Nascimento, Caroline da Silva Moraes, Rod James Dillon, Viv Maureen Dillon, Gisela Lara da Costa, Manoel Marques Evangelista Oliveira and Fernando Ariel Genta
Microbiol. Res. 2026, 17(6), 110; https://doi.org/10.3390/microbiolres17060110 - 5 Jun 2026
Viewed by 161
Abstract
Leishmaniases are vector-borne diseases transmitted by phlebotomine sand flies. While bacterial associations in sand fly microbiota are well studied, fungal communities remain poorly characterized, despite their potential role in insect biology and parasite transmission. This study aimed to isolate and characterize yeast-like and [...] Read more.
Leishmaniases are vector-borne diseases transmitted by phlebotomine sand flies. While bacterial associations in sand fly microbiota are well studied, fungal communities remain poorly characterized, despite their potential role in insect biology and parasite transmission. This study aimed to isolate and characterize yeast-like and filamentous fungi from different developmental stages of Lutzomyia longipalpis, the main vector of visceral leishmaniasis in the Americas, to expand knowledge on fungal microbiota and its possible relevance to vector–parasite interactions. Sand fly eggs, larvae, pupae, and adults were sampled from a laboratory colony. Fungi were isolated from insect tissues and diets using culture-based methods. Morphological identification was complemented by partial sequencing of the ITS1-5.8S-ITS2 rDNA region to identify the species. Four fungi were consistently recovered: Candida guilliermondii, Cutaneotrichosporon dermatis, Penicillium sp., and Aspergillus sp. Their presence varied across developmental stages. Presence in the gut was observed for Cu. dermatis, Penicillium sp. and Aspergillus sp. in larvae and C. guilliermondii in adult females. Evidence suggested their presence in different stages from larvae to pupae, and sex-specific differences in adults, with fungi detected only in females. This work documented the mycobiota that may be associated with L. longipalpis, including the first report of Cutaneotrichosporon in sand flies. These findings highlight fungi that may be potential modulators of sand fly biology and Leishmania development, warranting further investigation into their ecological and epidemiological roles. Full article
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15 pages, 20667 KB  
Article
First Report and Biological Characterization of Penicillium crustosum Causing Root Rot in Polygonatum kingianum (Yunnan, China)
by Ming-Xian Zhang, Zi-Han Chen, Li-Hua Wang, Xiao-Yi Yang, You-Yong Zhu and Yu Zhao
Plants 2026, 15(11), 1739; https://doi.org/10.3390/plants15111739 - 3 Jun 2026
Viewed by 203
Abstract
Root rot is a major disease restricting the cultivation and production of Polygonatum kingianum Coll. et Hemsl. This study aimed to identify the causal agent and characterize its biological properties. Pathogens were isolated from diseased rhizomes showing typical symptoms, and their pathogenicity was [...] Read more.
Root rot is a major disease restricting the cultivation and production of Polygonatum kingianum Coll. et Hemsl. This study aimed to identify the causal agent and characterize its biological properties. Pathogens were isolated from diseased rhizomes showing typical symptoms, and their pathogenicity was confirmed through Koch’s postulates using both detached rhizome inoculation and field pot experiments with spore suspension irrigation, in which typical root rot symptoms were reproduced. Based on morphological characteristics and multi-locus phylogenetic analysis (ITS, CaM, RPB2, and TUB), the pathogen was identified as Penicillium crustosum. Biological characterization revealed that the optimal conditions for mycelial growth and sporulation were 25 °C and pH 8–9, with Czapek agar being the most suitable medium. Light conditions significantly influenced fungal development; continuous darkness (24 h) favored mycelial growth, while an alternating light/dark cycle (12 h/12 h) significantly enhanced sporulation. Furthermore, the pathogen exhibited the highest utilization efficiency for soluble starch as a carbon source and peptone or yeast extract as a nitrogen source. These physiological traits suggest a strong adaptive capacity of the pathogen to environmental conditions associated with host rhizomes, which may contribute to disease development under cultivation conditions. To our knowledge, this is the first report of P. crustosum causing root rot in P. kingianum. The findings provide a basis for accurate pathogen identification and improve current understanding of the biological characteristics of this pathogen, thereby supporting future studies on disease monitoring and management. Full article
(This article belongs to the Special Issue Advances in Plant–Fungal Pathogen Interaction—2nd Edition)
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20 pages, 8970 KB  
Article
Data-Driven Identification of Favorable Multi-Fungal Inoculation Timing for Enhanced Humic Acid Recovery from Pretreated Crop Straws
by Peipei Zhang, Chao Zhao, Kunjie Chen, Lijun Xu, Farman Ali Chandio, Xiangjun Zhao and Bin Li
Agriculture 2026, 16(11), 1228; https://doi.org/10.3390/agriculture16111228 - 2 Jun 2026
Viewed by 220
Abstract
Humic acid (HA) production from crop straw is often limited by lignocellulosic recalcitrance and insufficient coordination among functional microorganisms. In this study, a data-driven strategy was developed to evaluate multi-fungal inoculation timing for HA recovery from pretreated straws. Three substrate platforms, namely raw [...] Read more.
Humic acid (HA) production from crop straw is often limited by lignocellulosic recalcitrance and insufficient coordination among functional microorganisms. In this study, a data-driven strategy was developed to evaluate multi-fungal inoculation timing for HA recovery from pretreated straws. Three substrate platforms, namely raw wheat straw (SW), steam-exploded corn straw (SC-SE), and ammoniated steam-exploded rice straw (SR-SE-N), were comparatively evaluated across an 81-run experimental matrix. Pretreatment markedly improved lignocellulose degradation and precursor turnover, with SR-SE-N showing the best humification performance. Based on the selected substrate, a two-factor interaction (2FI) model was established to describe the effects of inoculation timing on HA yield. The model was significant for HA prediction (R2 = 0.8768, adjusted R2 = 0.8398, predicted R2 = 0.7795). Inoculation timing strongly affected HA formation, and within the investigated timing range, the highest HA yield was obtained under simultaneous inoculation of Aspergillus niger, Phanerochaete chrysosporium, and Candida sp. Predicted and experimental HA yields were in close agreement, supporting the reliability of the model. These results indicate that favorable fungal inoculation timing is substrate-dependent and can be effectively identified through data-driven analysis within a bounded experimental range. The study provides a practical basis for improving HA biomanufacturing from pretreated agricultural residues. Full article
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20 pages, 13595 KB  
Article
Enhanced Activity of Leather Materials Coated with Silver, Copper and Graphene Oxides-Decorated TiO2 Nanocomposites and Gamma Irradiated
by Carmen Gaidau, Cosmin Alexe, Rodica Roxana Constantinescu, Laurentiu Dinca, Ioana Stanculescu, Mihalis Cutrubinis and Dragoș Cosma
Materials 2026, 19(11), 2358; https://doi.org/10.3390/ma19112358 - 2 Jun 2026
Viewed by 237
Abstract
It is known that antimicrobial surface treatments are one of the measures that can reduce the spread of viral, bacterial or fungal infections that threaten humans’ life. The excessive use of antibiotics has led to the emergence of resistant microorganisms. This study presents [...] Read more.
It is known that antimicrobial surface treatments are one of the measures that can reduce the spread of viral, bacterial or fungal infections that threaten humans’ life. The excessive use of antibiotics has led to the emergence of resistant microorganisms. This study presents the potential of nanocomposites to impart durable antimicrobial properties, which can be enhanced by the use of gamma radiation, even after two months of activation. Leather surfaces finished with composite titanium dioxide nanoparticles decorated with silver, copper oxide and/or graphene oxide were investigated before, immediately after, and after 60 days of gamma irradiation treatment. The antibacterial activity against Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 6538 was found to be maintained, and even slightly increased over time compared to unirradiated leathers. Analyses of the morphology and composition of the surface of treated leathers using SEM/EDS, ATR/FTIR, as well as photo activity tests allowed the identification of structural characteristics and the modifications induced by gamma radiation activation. Evaluating the resistance properties of leathers finished with the new nanoparticle composites compared to those finished classically confirmed the quality of the applied technologies. These results provide a solution for antimicrobial treatment of medical equipment, including footwear, with potential applications in other areas such as furniture, aircraft or car upholstery, clothing and bags. Full article
(This article belongs to the Special Issue Leather, Textiles and Bio-Based Materials)
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24 pages, 6565 KB  
Review
Bacterial Granulomatous Lung Diseases: Radiological Findings and Differential Diagnosis
by Picchi Stefano Giusto, Minieri Augusto, Lassandro Francesco, Russo Giuseppe and Lassandro Giulia
Infect. Dis. Rep. 2026, 18(3), 53; https://doi.org/10.3390/idr18030053 - 28 May 2026
Viewed by 155
Abstract
Background Granulomatous lung diseases include a spectrum of disorders, both infectious and noninfectious, unified by the presence of granulomas in the lung parenchyma. Granulomas are microscopic, organized collections of immune cells that arise as a response to persistent antigenic stimulation. Infectious granulomatous lung [...] Read more.
Background Granulomatous lung diseases include a spectrum of disorders, both infectious and noninfectious, unified by the presence of granulomas in the lung parenchyma. Granulomas are microscopic, organized collections of immune cells that arise as a response to persistent antigenic stimulation. Infectious granulomatous lung diseases arise from a variety of microbial agents, that include most frequently Mycobacterium tuberculosis, non-tuberculous mycobacteria, Nocardia, and Borrelia, as well as a wide range of fungal pathogens including Histoplasma, Cryptococcus, Pneumocystis, and Aspergillus species. Methods and Results: Definitive diagnosis is achieved through direct identification and subsequent culture of the causative pathogen in appropriate clinical specimens, including sputum, bronchoscopic samples, gastric aspirates, or pleural fluid. Imaging is fundamental for the detection and characterization of pulmonary granulomas. HRCT allows precise assessment of the number, size, and distribution of granulomatous lesions, can suggest an infectious etiology based on specific imaging patterns, and is essential for monitoring response to therapy over time. Differential diagnosis is challenging due to the numerous different imaging appearances with whom granulomatous lung diseases may manifest. Conclusions: The purpose of our review is to describe the spectrum of infectious granulomatous lung diseases caused by bacterial pathogens, highlighting their diverse radiologic presentations in order to assist radiologists in recognizing these entities and improving diagnostic accuracy. Full article
(This article belongs to the Section Bacterial Diseases)
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16 pages, 6572 KB  
Article
Unveiling Species Diversity Within Early-Diverging Fungi from China XIV: Five New Species of Mucorales
by Wen-Xiu Liu, Jing Zhu, Ning Wang, Heng Zhao, Xiao-Yong Liu and Zhi-Dong Zhang
J. Fungi 2026, 12(6), 386; https://doi.org/10.3390/jof12060386 - 27 May 2026
Viewed by 413
Abstract
Mucoralean fungi are mostly saprotrophic. During a fungal investigation of soil in Guangdong and Anhui provinces of China, five new species of Mucorales were discovered, namely Cunninghamella brevispora sp. nov., C. geminata sp. nov., Mucor chlamydosporiferus sp. nov., M. citrinus sp. nov., and [...] Read more.
Mucoralean fungi are mostly saprotrophic. During a fungal investigation of soil in Guangdong and Anhui provinces of China, five new species of Mucorales were discovered, namely Cunninghamella brevispora sp. nov., C. geminata sp. nov., Mucor chlamydosporiferus sp. nov., M. citrinus sp. nov., and M. magnisporus sp. nov. The identification is based on morphological characteristics, as well as molecular phylogenetics of the internal transcribed spacer (ITS), large subunit ribosomal RNA gene (LSU rDNA), translation elongation factor 1-alpha gene (TEF1α), and RNA polymerase II largest subunit gene (RPB1). Cunninghamella brevispora sp. nov. is sister to C. guizhouensis, and is distinguished by short sporangiophores. Cunninghamella geminata sp. nov. is sister to C. subclavata; rhizoids are absent in the former but well-developed in the latter. Mucor chlamydosporiferus sp. nov. is closely related to M. prayagensis, and is characterized by abundant chlamydospores. Mucor citrinus sp. nov. is closely related to M. paraorantomantidis, and is differentiated by pale yellow sporangiospores. Mucor magnisporus sp. nov. is sister to M. merdicola, and is discriminated by large sporangiospores. To date, with the addition of the five new species described herein, the total number of accepted species in the genus Cunninghamella and Mucor has increased to 49 and 163, respectively. Full article
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12 pages, 3198 KB  
Article
First Report of Leaf Spot of Spinacia oleracea Caused by Alternaria burnsii: Aerobiological Implications and Enzymatic Virulence Factor
by Tayyaba Afzal and Roshaan Ahmed
Aerobiology 2026, 4(2), 11; https://doi.org/10.3390/aerobiology4020011 - 26 May 2026
Viewed by 444
Abstract
Spinacia oleracea L. cultivation in South Asia is severely compromised by leaf spot disease caused by fungal plant pathogens, resulting in significant yield and quality losses. In this study, we report the first molecularly confirmed case of an Alternaria burnsii leaf spot on [...] Read more.
Spinacia oleracea L. cultivation in South Asia is severely compromised by leaf spot disease caused by fungal plant pathogens, resulting in significant yield and quality losses. In this study, we report the first molecularly confirmed case of an Alternaria burnsii leaf spot on S. oleracea in Pakistan. Symptomatic S. oleracea leaves exhibiting necrotic lesions with concentric rings were collected during a field survey across Bahawalpur district, Punjab, Pakistan in 2024. After isolation, purification and morphological identification it was identified that it belongs to the Alternaria genus. For the confirmation of species, molecular identification was performed; using the ITS and GAPDH primer revealed that the fungal plant pathogen causing leaf spot of S. oleracea is A. burnsii which was also confirmed by phylogenetic analysis. Koch’s postulates were carried out to confirm pathogenicity on detached leaf assays. To assess the virulence of A. burnsii enzymatic analysis was performed. Notably, enzymatic virulence profiling demonstrated a markedly increased production of polygalacturonase (PG: 16.0 ± 0.8 AU), pectin lyase (PNL: 12.0 ± 0.6 AU) and cellulase (CL: 14.0 ± 0.7 AU) relative to controls (all p < 0.001; LSD = 0.16), with PG having the greatest relative increase. This report expands the known host range for A. burnsii and highlights its two-fold threat: as a bioaerosol disseminable by wind and an enzymatic pathogen. These findings highlight the urgent need for integrated disease management strategies for suppressing leaf spot disease in S. oleracea agroecosystems. Full article
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29 pages, 8097 KB  
Article
Identification of GPI-Anchored Wall Transfer Protein 1 Modulators for Fungal Infections Through Generative AI and Physics-Based Approaches
by Ibrahim A. Alsarra, Rupesh Chikhale, Abdullah M. Al-Mohizea and Md Ataul Islam
Int. J. Mol. Sci. 2026, 27(11), 4767; https://doi.org/10.3390/ijms27114767 - 25 May 2026
Viewed by 314
Abstract
Glycosylphosphatidylinositol (GPI) anchored wall transfer protein 1 (GWT1), a fungal-specific inositol acyltransferase, catalyzes the palmitoylation of GlcN-PI in GPI-anchor biosynthesis, crucial for mannoprotein trafficking and attachment, which are vital for cell wall integrity, biofilm formation, and virulence. More than 60,000 AI-generated molecules produced [...] Read more.
Glycosylphosphatidylinositol (GPI) anchored wall transfer protein 1 (GWT1), a fungal-specific inositol acyltransferase, catalyzes the palmitoylation of GlcN-PI in GPI-anchor biosynthesis, crucial for mannoprotein trafficking and attachment, which are vital for cell wall integrity, biofilm formation, and virulence. More than 60,000 AI-generated molecules produced using REINVENT4 were screened using ADMET-AI and GNINA. DeepSA and PharmacoNet were used to select synthesizable and pharmacophorically rich molecules. The dynamic behaviour was explored using molecular dynamics (MD). Finally, molecular reactivity was assessed using density functional theory (DFT). After ADMET filtering, 6190 compounds were docked against GWT1, of which 315 showed better predicted binding energies than the co-crystal ligand. DeepSA identified 105 readily synthesizable candidates, and PharmacoNet retained 32 compounds with favourable pharmacophoric features, from which four final candidates (AF_M1, AF_M2, AF_M3, and AF_M4) were prioritized for further analysis. MD simulation suggested stable binding behavior towards GWT1. DFT analysis indicated favourable electronic properties, low HOMO-LUMO energy gaps, and stable optimized geometries. These molecules could serve as promising lead candidates and potential new therapeutic agents for invasive fungal infections, pending validation. Full article
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22 pages, 604 KB  
Article
Microbiota Characterization and Bioactive Potential of Broccoli (Brassica oleracea var. italica) By-Products: Implications for Sustainable Antimicrobial Applications
by Iris Gudiño, María José Benito, Alberto Martín and Rocío Casquete
Foods 2026, 15(10), 1786; https://doi.org/10.3390/foods15101786 - 18 May 2026
Viewed by 256
Abstract
Broccoli (Brassica oleracea var. italica) by-products represent an abundant and underutilized source of bioactive compounds with potential applications in sustainable food systems. This study aimed to characterize the microbiota associated with different plant fractions (leaves, stems, and heads) of broccoli (Parthenon [...] Read more.
Broccoli (Brassica oleracea var. italica) by-products represent an abundant and underutilized source of bioactive compounds with potential applications in sustainable food systems. This study aimed to characterize the microbiota associated with different plant fractions (leaves, stems, and heads) of broccoli (Parthenon and Tritón cultivars) and to evaluate the antioxidant and antimicrobial properties of their extracts, using cauliflower as a reference. Microbial counts and fungal identification (ITS sequencing) were performed, while phytochemical profiles were analyzed by HPLC-ESI-QTOF. Antioxidant activity was assessed using DPPH and ABTS assays, and antimicrobial activity under in vitro conditions was evaluated against selected foodborne bacteria and phytopathogenic fungi. Broccoli by-products, particularly leaves, showed lower microbial loads in certain cultivars and were rich in phenolic compounds and glucosinolates; however, higher phenolic content did not always correlate with greater antioxidant activity, highlighting the importance of compound composition. All extracts showed strong antibacterial activity at higher concentrations, especially against Listeria spp. Notably, antifungal activity was selective but relevant, with consistent inhibition observed against Alternaria alternata, while Penicillium purpurogenum and Botrytis cinerea exhibited higher resistance. Overall, these findings highlight the potential of broccoli by-products as sustainable sources of natural bioactive compounds for food applications, particularly in the development of preservation strategies and postharvest treatments. Further studies focusing on individual compounds and their specific biological activities are needed to better understand the mechanisms underlying these effects and to support their application in real food systems. Full article
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13 pages, 1988 KB  
Article
Near-Infrared Transmittance Spectroscopy for Early Screening of Alternaria Contamination and Alternariol Risk in Durum Wheat
by Alessandro Cammerata, Viviana Del Frate, Angela Iori and Francesco Gallucci
Agriculture 2026, 16(10), 1102; https://doi.org/10.3390/agriculture16101102 - 17 May 2026
Viewed by 370
Abstract
Early and non-destructive identification of fungal contamination in cereals is essential to support post-harvest management, reduce economic losses, and mitigate food safety risks along the wheat supply chain. Among filamentous fungi, Alternaria spp. are widespread contaminants of durum wheat and producers of toxic [...] Read more.
Early and non-destructive identification of fungal contamination in cereals is essential to support post-harvest management, reduce economic losses, and mitigate food safety risks along the wheat supply chain. Among filamentous fungi, Alternaria spp. are widespread contaminants of durum wheat and producers of toxic secondary metabolites such as alternariol (AOH), whose early detection remains analytically challenging. The aim of this study was to evaluate the potential of near-infrared transmittance (NIT) spectroscopy as a rapid, non-destructive pre-screening tool for the early identification of Alternaria-contaminated durum wheat lots and associated AOH risk. Samples from three durum wheat cultivars were artificially inoculated with Alternaria spp. and monitored over time. NIT spectra (570–1100 nm) were acquired in transmittance mode and analyzed using partial least squares (PLS) regression, focusing on the 870–1100 nm spectral region. Clear and time-dependent spectral differences were observed between inoculated and control samples, with the strongest discriminative features at 834 and 966 nm. Classification performance was high, with area under the curve (AUC) values between 0.96 and 0.97. ELISA analysis confirmed progressive AOH accumulation in inoculated kernels, consistent with the observed spectral changes, while control experiments excluded autoclaving and visual grain damage as confounding factors. From an applied perspective, the results indicate that NIT spectroscopy can support post-harvest decision-making as a rapid pre-screening approach, enabling the prioritization of suspect wheat lots for confirmatory analytical testing. Multivariate analysis further confirmed the consistency of spectral differences across datasets. Full article
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