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21 pages, 1458 KiB  
Article
Production of a Biosurfactant for Application in the Cosmetics Industry
by Ana Paula Barbosa Cavalcanti, Gleice Paula de Araújo, Káren Gercyane de Oliveira Bezerra, Fabíola Carolina Gomes de Almeida, Maria da Glória Conceição da Silva, Alessandra Sarubbo, Cláudio José Galdino da Silva Júnior, Rita de Cássia Freire Soares da Silva and Leonie Asfora Sarubbo
Fermentation 2025, 11(8), 451; https://doi.org/10.3390/fermentation11080451 (registering DOI) - 2 Aug 2025
Abstract
The cosmetics industry has been seeking to develop products with renewable natural ingredients to reduce the use of or even replace synthetic substances. Biosurfactants can help meet this demand. These natural compounds are renewable, biodegradable, and non-toxic or have low toxicity, offering minimal [...] Read more.
The cosmetics industry has been seeking to develop products with renewable natural ingredients to reduce the use of or even replace synthetic substances. Biosurfactants can help meet this demand. These natural compounds are renewable, biodegradable, and non-toxic or have low toxicity, offering minimal risk to humans and the environment, which has attracted the interest of an emerging consumer market and, consequently, the cosmetics industry. The aim of the present study was to produce a biosurfactant from the yeast Starmerella bombicola ATCC 22214 cultivated in a mineral medium containing 10% soybean oil and 5% glucose. The biosurfactant reduced the surface tension of water from 72.0 ± 0.1 mN/m to 33.0 ± 0.3 mN/m after eight days of fermentation. The yield was 53.35 ± 0.39 g/L and the critical micelle concentration was 1000 mg/L. The biosurfactant proved to be a good emulsifier of oils used in cosmetic formulations, with emulsification indices ranging from 45.90 ± 1.69% to 68.50 ± 1.10%. The hydrophilic–lipophilic balance index demonstrated the wetting capacity of the biosurfactant and its tendency to form oil-in-water (O/W) emulsions, with 50.0 ± 0.20% foaming capacity. The biosurfactant did not exhibit cytotoxicity in the MTT assay or irritant potential. Additionally, an antioxidant activity of 58.25 ± 0.32% was observed at a concentration of 40 mg/mL. The compound also exhibited antimicrobial activity against various pathogenic microorganisms. The characterisation of the biosurfactant using magnetic nuclear resonance and Fourier transform infrared spectroscopy revealed that the biomolecule is a glycolipid with an anionic nature. The results demonstrate that biosurfactant produced in this work has potential as an active biotechnological ingredient for innovative, eco-friendly cosmetic formulations. Full article
(This article belongs to the Special Issue The Industrial Feasibility of Biosurfactants)
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17 pages, 3272 KiB  
Review
Timing Is Everything: The Fungal Circadian Clock as a Master Regulator of Stress Response and Pathogenesis
by Victor Coca-Ruiz and Daniel Boy-Ruiz
Stresses 2025, 5(3), 47; https://doi.org/10.3390/stresses5030047 (registering DOI) - 1 Aug 2025
Abstract
Fungi, from saprophytes to pathogens, face predictable daily fluctuations in light, temperature, humidity, and nutrient availability. To cope, they have evolved an internal circadian clock that confers a major adaptive advantage. This review critically synthesizes current knowledge on the molecular architecture and physiological [...] Read more.
Fungi, from saprophytes to pathogens, face predictable daily fluctuations in light, temperature, humidity, and nutrient availability. To cope, they have evolved an internal circadian clock that confers a major adaptive advantage. This review critically synthesizes current knowledge on the molecular architecture and physiological relevance of fungal circadian systems, moving beyond the canonical Neurospora crassa model to explore the broader phylogenetic diversity of timekeeping mechanisms. We examine the core transcription-translation feedback loop (TTFL) centered on the FREQUENCY/WHITE COLLAR (FRQ/WCC) system and contrast it with divergent and non-canonical oscillators, including the metabolic rhythms of yeasts and the universally conserved peroxiredoxin (PRX) oxidation cycles. A central theme is the clock’s role in gating cellular defenses against oxidative, osmotic, and nutritional stress, enabling fungi to anticipate and withstand environmental insults through proactive regulation. We provide a detailed analysis of chrono-pathogenesis, where the circadian control of virulence factors aligns fungal attacks with windows of host vulnerability, with a focus on experimental evidence from pathogens like Botrytis cinerea, Fusarium oxysporum, and Magnaporthe oryzae. The review explores the downstream pathways—including transcriptional cascades, post-translational modifications, and epigenetic regulation—that translate temporal signals into physiological outputs such as developmental rhythms in conidiation and hyphal branching. Finally, we highlight critical knowledge gaps, particularly in understudied phyla like Basidiomycota, and discuss future research directions. This includes the exploration of novel clock architectures and the emerging, though speculative, hypothesis of “chrono-therapeutics”—interventions designed to disrupt fungal clocks—as a forward-looking concept for managing fungal infections. Full article
(This article belongs to the Collection Feature Papers in Plant and Photoautotrophic Stresses)
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14 pages, 6242 KiB  
Article
Characteristic Analysis of Ictalurus punctatus STING and Screening Validation of Interacting Proteins with Ictalurid herpesvirus 1
by Lihui Meng, Shuxin Li, Hongxun Chen, Sheng Yuan and Zhe Zhao
Microorganisms 2025, 13(8), 1780; https://doi.org/10.3390/microorganisms13081780 - 30 Jul 2025
Viewed by 110
Abstract
The innate immune response is an important defense against invading pathogens. Stimulator of interferon gene (STING) plays an important role in the cyclic GMP-AMP synthase (cGAS)-mediated activation of type I IFN responses. However, some viruses have evolved the ability to inhibit the function [...] Read more.
The innate immune response is an important defense against invading pathogens. Stimulator of interferon gene (STING) plays an important role in the cyclic GMP-AMP synthase (cGAS)-mediated activation of type I IFN responses. However, some viruses have evolved the ability to inhibit the function of STING and evade the host antiviral defenses. Understanding both the mechanism of action and the viruses targets of STING effector is important because of their importance to evade the host antiviral defenses. In this study, the STING (IpSTING) of Ictalurus punctatus was first identified and characterized. Subsequently, the yeast two-hybrid system (Y2HS) was used to screen for proteins from channel catfish virus (CCV, Ictalurid herpesvirus 1) that interact with IpSTING. The ORFs of the CCV were cloned into the pGBKT7 vector and expressed in the AH109 yeast strain. The bait protein expression was validated by autoactivation, and toxicity investigation compared with control (AH109 yeast strain transformed with empty pGBKT7 and pGADT7 vector). Two positive candidate proteins, ORF41 and ORF65, were identified through Y2HS screening as interacting with IpSTING. Their interactions were further validated using co-immunoprecipitation (Co-IP). This represented the first identification of interactions between IpSTING and the CCV proteins ORF41 and ORF65. The data advanced our understanding of the functions of ORF41 and ORF65 and suggested that they might contribute to the evasion of host antiviral defenses. However, the interaction mechanism between IpSTING, and CCV proteins ORF41 and ORF65 still needs to be further explored. Full article
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24 pages, 2240 KiB  
Article
Yeast Diversity on Sandy Lake Beaches Used for Recreation in Olsztyn, Poland
by Tomasz Bałabański, Anna Biedunkiewicz and Jan P. Jastrzębski
Pathogens 2025, 14(8), 744; https://doi.org/10.3390/pathogens14080744 - 29 Jul 2025
Viewed by 396
Abstract
Yeasts possess a range of environmental adaptations that allow them to colonize soil and sand. They can circulate seasonally between different components of lake ecosystems, including beach sand, water, and the coastal phyllosphere. The accumulation of people on beaches promotes the development and [...] Read more.
Yeasts possess a range of environmental adaptations that allow them to colonize soil and sand. They can circulate seasonally between different components of lake ecosystems, including beach sand, water, and the coastal phyllosphere. The accumulation of people on beaches promotes the development and transmission of yeasts, posing an increasing sanitary and epidemiological risk. The aim of this study was to determine the species and quantitative composition of potentially pathogenic and pathogenic yeasts for humans present in the sand of supervised and unsupervised beaches along the shores of lakes in the city of Olsztyn (northeastern Poland). The study material consisted of sand samples collected during two summer seasons (2019; 2020) from 12 research sites on sandy beaches of four lakes located within the administrative boundaries of Olsztyn. Standard isolation and identification methods used in diagnostic mycological laboratories were applied and are described in detail in the following sections of this study. A total of 259 yeast isolates (264, counting species in two-species isolates separately) belonging to 62 species representing 47 genera were obtained during the study. Among all the isolates, five were identified as mixed (two species from a single colony). Eight isolated species were classified into biosafety level 2 (BSL-2) and risk group 2 (RG-2). The highest average number of viable yeast cells was found in sand samples collected in July 2019 (5.56 × 102 CFU/g), August, and September 2020 (1.03 × 103 CFU/g and 1.94 × 103 CFU/g, respectively). The lowest concentrations were in samples collected in April, September, and October 2019, and October 2020 (1.48 × 102 CFU/g, 1.47 × 102 CFU/g, 1.40 × 102 CFU/g, and 1.40 × 102 CFU/g, respectively). The results indicate sand contamination with yeasts that may pose etiological factors for human mycoses. In light of these findings, continuous sanitary-epidemiological monitoring of beach sand and further studies on its mycological cleanliness are warranted, along with actions leading to appropriate legal regulations. Full article
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19 pages, 3813 KiB  
Article
An OSMAC Strategy for the Production of Antimicrobial Compounds by the Amazonian Fungi Talaromyces pinophilus CCM-UEA-F0414 and Penicillium paxilli CCM-UEA-F0591
by Cleudiane Pereira de Andrade, Caroline Dutra Lacerda, Raíssa Assímen Valente, Liss Stone de Holanda Rocha, Anne Terezinha Fernandes de Souza, Dorothy Ívila de Melo Pereira, Larissa Kirsch Barbosa, Cleiton Fantin, Sergio Duvoisin Junior and Patrícia Melchionna Albuquerque
Antibiotics 2025, 14(8), 756; https://doi.org/10.3390/antibiotics14080756 - 27 Jul 2025
Viewed by 267
Abstract
Background/Objectives: The emergence of antimicrobial resistance represents a critical global health threat, requiring the discovery of novel bioactive compounds. Fungi from Amazonian biodiversity are promising sources of secondary metabolites with potential antimicrobial activity. This study aimed to investigate the production of antimicrobial compounds [...] Read more.
Background/Objectives: The emergence of antimicrobial resistance represents a critical global health threat, requiring the discovery of novel bioactive compounds. Fungi from Amazonian biodiversity are promising sources of secondary metabolites with potential antimicrobial activity. This study aimed to investigate the production of antimicrobial compounds by two Amazonian fungal strains using the OSMAC (One Strain–Many Compounds) approach. Methods: Two fungal strains, Talaromyces pinophilus CCM-UEA-F0414 and Penicillium paxilli CCM-UEA-F0591, were cultivated under five distinct culture media to modulate secondary metabolite production. Ethyl acetate extracts were prepared and evaluated for antimicrobial activity against Gram-positive and Gram-negative bacteria, as well as pathogenic yeasts. Chemical characterization was performed using thin-layer chromatography (TLC), Fourier Transform Infrared Spectroscopy (FTIR), Ultraviolet–Visible (UV-Vis) spectroscopy, and Ultra-High-Performance Liquid Chromatography with Diode Array Detection (uHPLC-DAD). Results: The extracts exhibited significant antimicrobial activity, with minimum inhibitory concentrations (MICs) ranging from 78 to 5000 µg/mL. Chemical analyses revealed the presence of phenolic compounds, particularly caffeic and chlorogenic acids. Variations in the culture media substantially affected both the metabolite profiles and antimicrobial efficacy of the extracts. Conclusions: The OSMAC strategy effectively enhanced the metabolic diversity of the Amazonian fungal strains, leading to the production of bioactive metabolites with antimicrobial potential. These findings support the importance of optimizing culture conditions to unlock the biosynthetic capacity of Amazonian fungi as promising sources of antimicrobial agents. Full article
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17 pages, 1978 KiB  
Article
Insights into Persian Gulf Beach Sand Mycobiomes: Promises and Challenges in Fungal Diversity
by Abolfazl Saravani, João Brandão, Bahram Ahmadi, Ali Rezaei-Matehkolaei, Mohammad Taghi Hedayati, Mahdi Abastabar, Hossein Zarrinfar, Mojtaba Nabili, Leila Faeli, Javad Javidnia, Shima Parsay, Zahra Abtahian, Maryam Moazeni and Hamid Badali
J. Fungi 2025, 11(8), 554; https://doi.org/10.3390/jof11080554 - 26 Jul 2025
Viewed by 374
Abstract
Beach Sand Mycobiome is currently among the most important health challenges for viticulture in the world. Remarkably, the study of fungal communities in coastal beach sand and recreational waters remains underexplored despite their potential implications for human health. This research aimed to assess [...] Read more.
Beach Sand Mycobiome is currently among the most important health challenges for viticulture in the world. Remarkably, the study of fungal communities in coastal beach sand and recreational waters remains underexplored despite their potential implications for human health. This research aimed to assess the prevalence of fungal species and the antifungal susceptibility profiles of fungi recovered from the beaches of the Persian Gulf and the Sea of Oman. Sand and seawater samples from 39 stations distributed within 13 beaches along the coastline were collected between May and July 2023. The grown isolates were identified at the species level based on morphological characteristics and DNA sequencing. Antifungal susceptibility testing was performed according to the Clinical Laboratory Standards Institute guidelines. Of 222 recovered isolates, 206 (92.8%) filamentous fungi and 16 (7.2%) yeast strains were identified. Sand-recovered fungi comprised 82.9%, while water-originated fungi accounted for 17.1%. The DNA sequencing technique categorized 191 isolates into 13 genera and 26 species. The most recovered genus was Aspergillus (68.9%), and Aspergillus terreus sensu stricto was the commonly identified species (26.14%). Voriconazole was the most effective antifungal drug against Aspergillus species. Research on fungal contamination levels at these locations could provide a foundation for establishing regulatory frameworks to diminish fungal risks, thereby enhancing public health protection. The ecological significance of fungal communities in sandy beaches to human infections remains to be explored, and earlier reports in the literature may motivate researchers to focus on detecting this mycobiome in natural environments where further investigation is warranted. Ultimately, our discovery serves as a reminder that much remains to be learned about pathogenic fungi and underscores the need for vigilance in areas where emerging pathogens have not yet been identified. Full article
(This article belongs to the Special Issue Fungi Activity on Remediation of Polluted Environments, 2nd Edition)
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11 pages, 971 KiB  
Case Report
Gastric Candidiasis in Five Horses: A Case Series
by Patricia Neira-Egea, Clara Alamar Malvoisin, María de la Cuesta-Torrado, Claudia Bautista-Erler, Valentina Vitale, Sandra Jolly and Carla Cesarini
Microorganisms 2025, 13(8), 1746; https://doi.org/10.3390/microorganisms13081746 - 25 Jul 2025
Viewed by 262
Abstract
Candida spp. are ubiquitous yeasts that are part of most mammals’ microbiota and can become opportunistic pathogens under predisposing conditions. Interestingly, recent studies in human medicine report an increased abundance of Candida spp. in association with inflammatory bowel disease (IBD). Gastrointestinal candidiasis has [...] Read more.
Candida spp. are ubiquitous yeasts that are part of most mammals’ microbiota and can become opportunistic pathogens under predisposing conditions. Interestingly, recent studies in human medicine report an increased abundance of Candida spp. in association with inflammatory bowel disease (IBD). Gastrointestinal candidiasis has been primarily reported in neonatal foals, but not in adult horses. The aim of this study is to describe the morphological, histopathological, and microbiological features of gastric lesions associated with Candida infiltration in five horses referred to two tertiary hospitals for different reasons. Clinical features, findings from gastroscopy, gastric, and duodenal biopsies, as well as fungal and bacterial cultures obtained from gastric lesions will be reported. Macroscopically, gastric lesions showed a characteristic yellow/white pseudo-membranous appearance, similar to lesions reported in foals. The presence of Candida spp. was confirmed by positive culture and/or histopathological evidence of fungal infiltration on the gastric epithelium. Three out of five horses showed histopathological changes in duodenal biopsies, potentially suggesting IBD. These results demonstrate that gastric candidiasis can occur in adult horses. Further research is needed to elucidate the pathogenesis, predisposing factors, and clinical relevance of Candida spp. infections in the equine stomach, as well as their potential impact on gastrointestinal health and overall performance. Full article
(This article belongs to the Section Veterinary Microbiology)
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12 pages, 467 KiB  
Review
Exophiala Bloodstream Infections in Humans—A Narrative Review
by Afroditi Ziogou, Alexios Giannakodimos, Ilias Giannakodimos, Stella Baliou, Andreas G. Tsantes and Petros Ioannou
Pathogens 2025, 14(7), 706; https://doi.org/10.3390/pathogens14070706 - 17 Jul 2025
Viewed by 323
Abstract
Background: Exophiala spp. are dematiaceous fungi with opportunistic pathogenic potential and a widespread environmental presence. Clinical cases of Exophiala spp. fungemia are uncommon. Although rarely encountered in the general population, these organisms are increasingly reported in immunocompromised individuals or those with complex [...] Read more.
Background: Exophiala spp. are dematiaceous fungi with opportunistic pathogenic potential and a widespread environmental presence. Clinical cases of Exophiala spp. fungemia are uncommon. Although rarely encountered in the general population, these organisms are increasingly reported in immunocompromised individuals or those with complex underlying health conditions. Objectives: This review seeks to examine all documented human cases of Exophiala spp. fungemia, with particular focus on aspects such as epidemiology, microbiological features, resistance patterns, therapeutic approaches and associated mortality rates. Methods: A narrative review was conducted using data sourced from the PubMed/MedLine and Scopus databases. Results: A total of 19 articles described infections in 32 patients involving Exophiala spp. fungemia. The mean patient age was 49.2 years, and 65.6% were male. Central venous catheters emerged as the leading predisposing factor (96.9%). Fever represented the most frequent clinical presentation (50%), followed by organ dysfunction (21.9%). The yeast generally demonstrated susceptibility to voriconazole and itraconazole. Voriconazole was also the most frequently administered antifungal (62.5%), followed by amphotericin (31.3%) and micafungin (28.1%). Overall mortality reached 34.4%, with 25% of deaths specifically caused by the infection. Conclusions: Given the potential of Exophiala spp. to cause severe fungemia, healthcare professionals, particularly clinicians and microbiologists, should consider this pathogen in the differential diagnosis when black yeast is detected in blood cultures, especially in patients with immunodeficiency or significant comorbidities, to ensure timely and accurate identification. Full article
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26 pages, 1399 KiB  
Review
Clinical and Molecular Advances on the Black Yeast Exophiala dermatitidis
by Jakub Suchodolski, Mateusz Parol, Karolina Pawlak, Agata Piecuch and Rafał Ogórek
Int. J. Mol. Sci. 2025, 26(14), 6804; https://doi.org/10.3390/ijms26146804 - 16 Jul 2025
Viewed by 394
Abstract
Exophiala dermatitidis is an emerging black yeast recognized in both superficial and life-threatening infections, including those in immunocompetent hosts. This narrative review focuses on recent developments (mostly between 2019 and 2025) in two major areas. First, we examined the clinical and epidemiological background [...] Read more.
Exophiala dermatitidis is an emerging black yeast recognized in both superficial and life-threatening infections, including those in immunocompetent hosts. This narrative review focuses on recent developments (mostly between 2019 and 2025) in two major areas. First, we examined the clinical and epidemiological background of E. dermatitidis, with particular focus on its involvement in cystic fibrosis and CARD9 deficiency, as well as central nervous system, ocular, and systemic infections. Second, we address the molecular basis of its pathogenicity, with particular attention to melanin production, capsule formation, and metabolic adaptation. We also discuss diagnostic challenges and antifungal susceptibility, highlighting gaps between laboratory studies and clinical outcomes. Full article
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15 pages, 2488 KiB  
Article
Antimicrobial Peptides Act-6 and Act 8-20 Derived from Scarabaeidae Cecropins Exhibit Differential Antifungal Activity
by Melissa Rodríguez, Lily J. Toro, Carolina Firacative, Beatriz L. Gómez, Bruno Rivas-Santiago, David Andreu, Jhon C. Castaño, German A. Téllez and Julián E. Muñoz
J. Fungi 2025, 11(7), 519; https://doi.org/10.3390/jof11070519 - 12 Jul 2025
Viewed by 373
Abstract
The number of fungal infections is steadily increasing, with considerable morbidity and mortality. Additionally, antifungal resistance is a growing concern, highlighting the need to develop new treatment options. One alternative is the use of antimicrobial peptides (AMPs). The aim of this study was [...] Read more.
The number of fungal infections is steadily increasing, with considerable morbidity and mortality. Additionally, antifungal resistance is a growing concern, highlighting the need to develop new treatment options. One alternative is the use of antimicrobial peptides (AMPs). The aim of this study was to assess the in vitro and in vivo antifungal activity of designed short AMPs, Act-6 and Act 8-20, derived from cecropin transcripts of beetles from the family Scarabaeidae, against eight reference strains of the pathogenic yeasts Candida and Cryptococcus. We also evaluated the effect of these modified AMPs on the biofilm, morphogenesis, and cell morphology of Candida albicans, as well as the in vivo activity via a murine model of disseminated candidiasis. The AMPs herein analyzed exhibit differential antifungal activity against the yeasts assessed, and inhibit biofilm, hyphae, and pseudohyphae formation with morphological alterations in C. albicans. Moreover, the fungal load in mice treated with these AMPs significantly decreased. Altogether, our results suggest that Act-6 and Act 8-20 are promising antifungal molecules to control mycoses. Full article
(This article belongs to the Special Issue New Strategies to Combat Human Fungal Infections)
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15 pages, 4602 KiB  
Article
Antifungal Nanocomposites from Honeybee Chitosan and Royal Jelly-Mediated Nanosilver for Suppressing Biofilm and Hyphal Formation of Candida albicans
by Mousa Abdullah Alghuthaymi
Polymers 2025, 17(14), 1916; https://doi.org/10.3390/polym17141916 - 11 Jul 2025
Viewed by 358
Abstract
Candida albicans complications challenged researchers and health overseers to discover effectual agents for suppressing such yeast growth, biofilm formation and conversion to hyphal form. The nanomaterials and their composites provided extraordinary bioactivities and functionalities as antimicrobial preparations. The extraction of chitosan (BCt) from [...] Read more.
Candida albicans complications challenged researchers and health overseers to discover effectual agents for suppressing such yeast growth, biofilm formation and conversion to hyphal form. The nanomaterials and their composites provided extraordinary bioactivities and functionalities as antimicrobial preparations. The extraction of chitosan (BCt) from honeybee corpuses was achieved as an innovative biopolymer for nanocomposite formation. The green (bio)synthesis of nanosilver (AgNPs) was promisingly performed using royal jelly (RJ) as a mediator of synthesis. The RJ-synthesized AgNPs had an average diameter of 3.61 nm and were negatively charged (−27.2 mV). The formulated nanocomposites from BCt/RJ/AgNPs at 2:1 (F1), 1:1 (F2), and 1:2 (F3) ratios had average diameters of 63.19, 27.65, and 52.74 nm, where their surface charges were +33.8, +29.3, and −11.5 mV, respectively. The infrared (FTIR) analysis designated molecules’ interactions, whereas the transmission microscopy emphasized the homogenous distribution and impedance of AgNPs within the biopolymers’ nanocomposites. Challenging C. albicans strains with nanomaterials/composites pinpointed their bioactivity for suppressing yeast growth and biofilm formation; the F2 nanocomposite exhibited superior actions, with the lowest inhibitory concentrations (MICs) of 125–175 mg/L, whereas the MIC ranges were 150–200 and 175–225 mg/L for F3 and F1, respectively. The different BCht/RJ/AgNP nanocomposites could entirely suppress the biofilm formation of all C. albicans strains. The scanning microscopy reflected the nanocomposite efficiency for C. albicans cell destruction and the complete suppression of hyphal formation. The application of generated BCht/RJ/AgNP nanocomposites is strongly recommended as they are effectual, natural and advanced materials for combating C. albicans pathogens. Full article
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13 pages, 1650 KiB  
Article
Isolation of a Monoclonal Human scFv Against Cytomegalovirus pp71 Antigen Using Yeast Display
by Kazuhisa Aoki, Rikio Yabe, Sayaka Ono, Mayumi Saeki, Yuri Tanno and Hidetaka Tanno
Antibodies 2025, 14(3), 57; https://doi.org/10.3390/antib14030057 - 10 Jul 2025
Viewed by 340
Abstract
Background: Human cytomegalovirus (CMV) is a major pathogen that poses significant risks to immunocompromised individuals and neonates. The tegument protein pp71, encoded by the UL82 gene, plays a pivotal role in initiating viral lytic replication and evading host immune responses. Despite its clinical [...] Read more.
Background: Human cytomegalovirus (CMV) is a major pathogen that poses significant risks to immunocompromised individuals and neonates. The tegument protein pp71, encoded by the UL82 gene, plays a pivotal role in initiating viral lytic replication and evading host immune responses. Despite its clinical relevance, standardized monoclonal antibodies (mAbs) for pp71 remain limited, prompting the need to expand the available repertoire of antibodies targeting this critical protein. Methods: In this study, we constructed a diverse human single-chain variable fragment (scFv) library using RNA derived from the B cells of four healthy donors. The library was expressed in Saccharomyces cerevisiae, and iterative rounds of magnetic-activated cell sorting (MACS) were performed against recombinant pp71. Clonal enrichment was monitored using flow cytometry. Results: Among the isolated clones, one designated ID2 exhibited high sensitivity and specificity for pp71, as demonstrated by flow cytometry, immunofluorescence, an enzyme-linked immunosorbent assay (ELISA), and biolayer interferometry (BLI). Conclusions: Collectively, these findings establish a novel pp71-specific mAb and underscore the utility of yeast surface display combined with MACS for expanding the antibody toolkit available for CMV research and diagnostics. Full article
(This article belongs to the Section Antibody Discovery and Engineering)
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13 pages, 1536 KiB  
Communication
Complete Sequence Analysis of Grapevine Leafroll-Associated Virus 4 and Interactions Between the Encoded Proteins
by Tingting Du, Yuxin Hao, Jie Gao, Shane Qiao, Guojun Hu, Fang Ren, Xudong Fan and Yafeng Dong
Viruses 2025, 17(7), 952; https://doi.org/10.3390/v17070952 - 5 Jul 2025
Viewed by 402
Abstract
Grapevine leafroll disease is one of the most devastating diseases in the global viticulture industry. Grapevine leafroll-associated virus 4 is one of the main pathogens causing this disease. In this study, we obtained the complete genome sequences of two Chinese isolates of GLRaV-4 [...] Read more.
Grapevine leafroll disease is one of the most devastating diseases in the global viticulture industry. Grapevine leafroll-associated virus 4 is one of the main pathogens causing this disease. In this study, we obtained the complete genome sequences of two Chinese isolates of GLRaV-4 from ‘Baisainie’ and ‘Fantasy Seedless’ grapevines through high-throughput sequencing and overlapping RT-PCR combined with RACE technology. The sequences contain 13,814 and 13,824 nucleotides and code six open reading frames, respectively. Phylogenetic trees based on the coat protein (CP) and heat shock protein 70 (HSP70) genes show that in addition to other GLRaV-4 strains (strains 5, 6, 9, Pr, and Car), the GLRaV-4 strains were divided into two distinct groups. The two isolates obtained in this study were classified into separate branches within GLRaV-4 Group 1. Additionally, we systematically investigated the interactions between the proteins encoded by GLRaV-4 using the yeast two-hybrid and bimolecular fluorescence complementation techniques. We found significant interactions between the GLRaV-4-encoded p23 and HSP70 and CP. This study first reports the complete genomes of two different GLRaV-4 isolates from China and suggests that p23 protein encoded by GLRaV-4 may play an important role in viral pathogenicity due to its interactions with the other two proteins. Full article
(This article belongs to the Special Issue Diversity and Coinfections of Plant or Fungal Viruses, 3rd Edition)
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23 pages, 4022 KiB  
Article
Comprehensive Analysis of Bacterial Communities and Microbiological Quality of Frozen Edible Insects
by Sasiprapa Krongdang, Nipitpong Sawongta, Jintana Pheepakpraw, Achirawit Ngamsomchat, Sutee Wangtueai, Jittimon Wongsa, Thanya Parametthanuwat, Narin Charoenphun and Thararat Chitov
Foods 2025, 14(13), 2347; https://doi.org/10.3390/foods14132347 - 1 Jul 2025
Viewed by 395
Abstract
Edible insects are gaining traction worldwide; however, the existing data regarding their microbiological quality remain inadequate. This study investigated the bacterial communities and microbiological quality of five types of frozen edible insects commercially available in Thailand. Amplicon sequencing revealed Firmicutes (Bacillota) and Proteobacteria [...] Read more.
Edible insects are gaining traction worldwide; however, the existing data regarding their microbiological quality remain inadequate. This study investigated the bacterial communities and microbiological quality of five types of frozen edible insects commercially available in Thailand. Amplicon sequencing revealed Firmicutes (Bacillota) and Proteobacteria (Pseudomonadota) as the main phyla across all samples; Bacteroidota was predominant in house crickets, Actinobacteriota in silkworms, and Desulfobacterota was exclusively found in house and mole crickets. Culture-based assays showed total viable counts, lactic acid bacteria, yeasts–molds, and spore-formers ranging from 3.41–6.58, 2.52–7.41, 1.83–5.62, to 2.00–4.70 log CFU·g−1, respectively. In some samples, Enterobacteriaceae and Escherichia coli, key hygiene indicators, reached 5.05 and 2.70 log CFU·g−1, respectively. Among foodborne pathogens, presumptive Bacillus cereus was found to vary from <1.70 to 3.93 log CFU·g−1, while Clostridium perfringens and Staphylococcus aureus were under the quantitation limit, and Salmonella was absent. Overall, the results indicate significant variation in microbial diversity and quality among different insect types. The high levels of microbial hygiene indicators and foodborne pathogens in some samples raised food safety concerns and point to the need to develop or implement production guidelines and microbiological criteria for frozen edible insects to ensure food safety. Full article
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19 pages, 2039 KiB  
Article
The Study of Metschnikowia pulcherrima E1 in the Induction of Improved Gray Spot Disease Resistance in Loquat Fruit
by Xiaoya Li, Kunkun Wu, Xin Li, Yuhao Zhao and Weihong Sun
J. Fungi 2025, 11(7), 497; https://doi.org/10.3390/jof11070497 - 30 Jun 2025
Viewed by 427
Abstract
In this study, the dominant pathogenic fungus of gray spot disease in loquat, which was isolated from postharvest decaying loquat fruits in Zhenjiang, was identified as Pestalotiopsis vismiae (P. vismiae) by morphological characteristics and DNA sequencing. At the same time, a [...] Read more.
In this study, the dominant pathogenic fungus of gray spot disease in loquat, which was isolated from postharvest decaying loquat fruits in Zhenjiang, was identified as Pestalotiopsis vismiae (P. vismiae) by morphological characteristics and DNA sequencing. At the same time, a strain of yeast E1, which could effectively inhibit the pathogen, was isolated from the loquat leaves and soil and identified as Metschnikowia pulcherrima (M. pulcherrima) by morphological and molecular biological characteristics. It significantly reduced the natural decay of loquat fruits without affecting fruit quality. Metschnikowia pulcherrima E1 (M. pulcherrima E1) exhibited significant biocontrol efficacy against P. vismiae, the causal agent of gray spot in loquat, reducing disease incidence to 22.73% compared to 100% in the control group. Transcriptome analysis revealed 1444 differentially expressed genes (DEGs) (1111 upregulated, 333 downregulated), with key genes (CML19, XTH23, GSTU10) validated by RT-qPCR. The Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis highlighted enrichment in plant–pathogen interactions, glutathione metabolism, and phenylpropanoid biosynthesis. These findings provided molecular insights into yeast-induced resistance, bridging biocontrol applications with mechanistic studies. Full article
(This article belongs to the Special Issue Biological Control of Fungal Plant Pathogens)
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