Search Results (58)

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

Penicillium species are major postharvest pathogens of fruits and vegetables, causing significant economic losses and posing serious threats to food safety through mycotoxin contamination. This review systematically summarizes the pathogenic mechanisms, metabolic diversity, and eco-friendly strategies of postharvest Penicillium pathogens. The application of CRISPR-Cas9 technology has enabled precise functional analysis of pathogenicity-related genes (e.g., PacC, PeStuA) and regulatory elements involved in fungicide resistance (e.g., FlbC). RNA interference-based strategies, including host-induced gene silencing (HIGS) and spray-induced gene silencing (SIGS), offer promising non-transgenic approaches for disease control. Additionally, artificial intelligence-assisted species identification and fermentation regulation have improved research efficiency. Future integration of multidisciplinary technologies will facilitate sustainable management of postharvest diseases. Full article

Climate change is intensifying ecological instability in agricultural ecosystems, altering pest distribution and increasing the risk of fungal contamination and mycotoxin accumulation in crops. Among emerging threats, the invasive brown marmorated stink bug Halyomorpha halys has rapidly expanded across Europe, including Croatia. The species is polyphagous, and in the ripening of most plant crops, it accounts for a significant population and causes extensive plant damage. This study investigates the capacity of H. halys to disseminate mycotoxigenic fungi, with an emphasis on aflatoxin B₁ (AFB₁) and ochratoxin A (OTA) producers. One hundred adult H. halys individuals were collected from maize ears in Osijek, Croatia, in August 2023. Fungal propagules washed from their surfaces were inoculated on PDA and MEA media. A total of 212 pure fungal cultures were obtained, of which 202 belonged to the genera Aspergillus and Penicillium. Molecular identification using β-tubulin (BenA) gene sequencing revealed six Aspergillus and six Penicillium species, forming two well-supported phylogenetic clades. Aspergillus parasiticus was the most dominant species. Mycotoxin screening confirmed AFB₁ production in 20 isolates, all identified as A. parasiticus, while no OTA-producing strains were detected. The predominance of aflatoxigenic species on H. halys indicates that it may play a previously underrecognized role in the dispersal of mycotoxigenic fungi in agroecosystems. Full article

Patulin is a mycotoxin produced mainly by Penicillium expansum on apples. P. expansum is a fruit pathogen that can cause apple soft rot. However, much is unknown about the characteristics of P. expansum and influence of major environmental parameters on its patulin production and growth on apple puree agar media (APAM). In this study, we evaluated the influence of pH, temperature, and relative humidity (RH) on patulin production by P. expansum OM1 and its growth (colony diameter and mycelial dry weight) on APAM after isolation and identification of the patulin-producing fungal strain from an apple. The fungal isolate produced the largest quantity of patulin on APAM under 15 °C, pH 4.0, and RH 98%, while it had the highest growth rates on the same media under 25 °C, pH 4.0–6.0, and RH 98%. Our data demonstrated that three important physicochemical factors (pH, temperature, and RH) substantially influenced the patulin production by the fungal species and its growth on APAM. Moreover, our results revealed that patulin was not detected on APAM at 5 °C after 7 days of incubation and that a trace amount of patulin was produced by the fungal strain along with its slow growth on the same media at 5 °C after 14 days. It suggests that patulin contamination by P. expansum on apples could be controlled during postharvest storage below 5 °C. These findings could provide fundamental knowledge for development of efficient strategies to prevent the occurrence of apples contaminated with patulin produced by P. expansum on them during postharvest storage. Full article

Black pepper (Piper nigrum L.) has historically been among the most consumed spices globally. Brazil is one of the world’s largest producers and exporters, and is the largest in the Western Hemisphere. This study describes two new fungal species associated with black pepper commercialized in Brazil. The first, Penicillium pipericola sp. nov., belongs to the subgenus Penicillium, section Paradoxa, series Atramentosa. The second, Syncephalastrum brasiliense sp. nov., belongs to the order Mucorales, family Syncephalastraceae. The taxonomic classification of these species was supported by a pluralistic approach, based on multilocus phylogenetic analyses, morphological analyses, and metabolomics. Furthermore, the metabolomic analysis revealed considerable biosynthetic versatility of the new species under different cultivation conditions, producing metabolites with therapeutic and biotechnological potential. The identification of these species increases the understanding of fungal diversity in the black pepper production chain and may have important implications for the microbiological quality of the product, for the understanding of ecological interactions within the agroecosystem and for potential industrial applications. Full article

The genus Agave (family Asparagaceae) represents the second-most important group of plants in Mexico. Several fungal species have been identified as causal agents of leaf spot disease affecting Agave salmiana and A. lechuguilla, producing necrotic lesions that compromise plant health and productivity. Pathogenicity experiments were conducted under greenhouse conditions, field tests were performed, and in vitro antagonism using Trichoderma asperellum strain JEAB02 against selected pathogenic isolates was evaluated. Phylogenetic analysis of genomic DNA fragments allowed the identification of 26 fungal isolates belonging to Curvularia lunata, C. verruculosa, Bipolaris zeae, Alternaria alternata, Fusarium lactis, Epicoccum sorghinum, Myrmaecium rubricosum, Penicillium diversum, and Aspergillus oryzae. In pathogenicity assays under greenhouse conditions on A. salmiana and A. lechuguilla, treatments T5–T12 exhibited statistically similar levels of disease severity (33.10–37.29%), caused mainly by C. verruculosa, A. alternata, B. zeae, and F. lactis. In field tests, Agave plants inoculated with the selected pathogenic fungi (T4, T5, T7, T8, T10, and T11) showed 21.07–36.73% leaf damage after 75 days. The antagonistic effect of T. asperellum JEAB02 caused complete (100%) growth inhibition of the pathogenic isolate JCPN27 and inhibition levels from 99.81 to 99.98% for isolates JCPN18, JCPN24, JCPN28, JCPN29, JCPN31, and JCPN33, demonstrating its high potential as a biological control agent. Full article

Tannases (tannin acyl hydrolases, EC 3.1.1.20) are enzymes of industrial interest due to their ability to hydrolyze hydrolyzable tannins into bioactive compounds like gallic acid. In this study fungal strains capable of producing extracellular tannase were isolated and identified. From tannin-rich substrates, 24 fungal isolates were obtained, of which 17 showed tannase activity. Molecular identification based on calmodulin gene sequencing identified three species of tannase-producing black aspergilli: Aspergillus luchuensis, A. niger (formerly A. welwitschiae), and A. uvarum. The isolate A. uvarum HT4 exhibited the highest extracellular tannase activity (182 U/mL) and was selected for further study. Whole-genome sequencing of HT4 revealed 15 putative tannase genes, most sharing high identity with A. uvarum CBS 121591. Two divergent genes appeared to be acquired via horizontal gene transfer from Aspergillus brunneoviolaceus and Penicillium angulare. Proteomic analysis of the secretome confirmed the expression of two extracellular tannases. The enzyme showed optimal activity at pH 5.0–6.0 and 40–50 °C. Secretome analysis revealed hydrolytic enzymes typical of saprophytic fungi in lignocellulose-rich environments. Importantly, no biosynthetic gene clusters of major mycotoxins were detected, supporting the biosafety of HT4 for industrial applications. Full article

Caves and other subterranean ecosystems are characterized by stable, low temperatures, high humidity, and limited nutrient input, creating unique environments for extremophilic microorganisms. Among them, fungi play key roles in organic matter degradation, mineral interactions, and biogeochemical cycling, yet the diversity and adaptations of cold-adapted fungi in cave habitats remain insufficiently explored. This study investigated psychrophilic and psychrotolerant fungi inhabiting the stone surfaces of Ravništarka Cave in Eastern Serbia. Biofilm samples were collected from nine sites and analyzed using culture-based isolation on both nutrient-rich and diluted media, followed by incubation at 10 °C, 25 °C, and 37 °C. Fungal identification combined morphological characteristics with molecular analyses of the ITS region and BenA gene, while ecological roles were assigned using FUNGuild. A total of 41 fungal species were documented, spanning Ascomycota (53.1%), Basidiomycota (43.7%), and Mortierellomycota (3.1%) phyla. The genus Penicillium exhibited the greatest species richness, with 14 taxa documented, including P. chrysogenum, P. glandicola, and P. solitum, all previously associated with cold or oligotrophic environments. The psychrotolerant species Mortierella alpina was the only representative of Mortierellomycota. Ecological guild assignment revealed fungi functioning at different trophic levels, highlighting their multifunctional ecological roles in extreme subterranean habitats. Full article

Pseudomonas aeruginosa is a harmful pathogen that causes a variety of acute and chronic infections through quorum sensing (QS) mechanisms. The increasing resistance of this bacterium to numerous antibiotics has created a demand for new medications that specifically target QS. Endophytes can be the source of compounds with antibacterial properties. This research is the first to examine tannic acid (TA) produced by endophytic fungus as a potential biotherapeutic agent. A novel endophytic fungal isolate identified as Penicillium oxalicum was derived from the cladodes of Opuntia ficus-indica (L.). The species identification for this isolate was confirmed through sequencing of the internal transcribed spacer region. The metabolites from the culture of this isolate were extracted using ethyl acetate, then separated and characterized using chromatographic methods. This led to the acquisition of TA, a compound that shows strong anti-QS and excellent antibacterial effects against extensively drug-resistant P. aeruginosa strains. Furthermore, it was shown that treating P. aeruginosa with the obtained TA reduced the secretion of virulence factors controlled by QS in a dose-dependent manner, indicating that TA inhibited the QS characteristics of P. aeruginosa. Simultaneously, TA significantly inhibited the expression of genes associated with QS, including rhlR/I, lasR/I, and pqsR. In addition, in silico virtual molecular docking showed that TA could efficiently bind to QS receptor proteins. Our results showed that P. oxalicum could be a new source of TA for the treatment of infections caused by extensively drug-resistant P. aeruginosa. Full article

This study aimed to examine fungi diversity in dried beef meat sold in Ekiti State, characterize the isolated fungi, and determine the aflatoxin-producing ability of the Aspergillus fungi in the samples. Dried beef meat was collected from different markets in Ekiti State and screened for the presence of filamentous fungi using molecular methods. Samples were cultured aseptically on potato dextrose agar (PDA) for fungi isolation, and molecular identification was performed using DNA extraction, Polymerase chain Reaction (PCR), ITS-1/ITS-4 primer pair, and nucleotide sequencing. The results obtained indicated a range of filamentous fungi genera including Aspergillus, Rhizopus, Penicillium, Fusarium, Cladosporium, Alternaria, and other fungi species contaminating the dried meat at (43%), (42%), (3%), (2%), (2%), (1%), and (7%), respectively. High incidences were recorded for Aspergillus flavus, Aspergillus niger, and Aspergillus fumigatus in most of the screened samples. Aspergillus flavus accounted for (24.7%) of all the Aspergillus species isolated with the presence of the gene needed for aflatoxin production. The occurrences of these filamentous fungal species pose a cause for concern, as most of these fungal species are known producers of certain toxic substances. Maximum likelihood phylogenetic analysis showed a high similarity index score, which indicated a good relationship between isolated Aspergillus Species and the closely related strains from GenBank, isolated from different sources and countries. The implication of this study is that consumer health may be at risk through exposure to contaminated dried meat. Full article

The European sweet cherry Prunus avium (L.), a member of the Rosaceae family, is one of the most popular and economically valuable fruits. However, the rapid spread of gummosis and poor management practices have become the major obstacles to their production. To identify pathogenic microorganisms responsible for gummosis disease, we conducted observations comparing the garden of Bailuyuan, which heavily suffered from gummosis disease and horn beetle damage, with the orchard of Mayuhe, which only suffered from gummosis disease, both from Xi’an, Shaanxi, China. Samples were obtained from the healthy tissues and gummosis disease tissues that used the Illumina sequence of 16S rRNA and the internal transcribed spacer region (ITS) to identify bacterial and fungal communities in these samples. An alpha diversity analysis revealed a significantly higher fungal diversity of disease than in healthy tissue in the gummosis period. The results suggested that an imbalance in the fungal genera may be associated with gummosis disease. Species relative analyses showed some bacterial genera (Pelagibacterium, Halomonas, Azospirillum, Aquabacterium and Alistipes) and fungal genera (Penicillium, Alternaria and Rhodotorula) in the diseased tissues of gummosis. Among these, the increased relative abundance of the bacteria genes Halomonas, Pelagibacterium, Chelativorans, Pantoea, Aquabacterium, Alternaria and fungi genes Penicillium, Cystobasidium, Rhodotorula may be associated with gummosis of P. avium. The bacterial genera Methylobacterium, Psychroglaciecola, Aeromonas, Conexibacter and fungal genera Didymella, Aureobasidium, Mycosphaerella, Meyerozyma are probably antagonists of the pathogen of gummosis. These findings are an initial step in the identification of potential candidates for the biological control of the disease. Full article

The Penicillium genus exhibits a broad global distribution and holds substantial economic value in sectors including agriculture, industry, and medicine. Particularly in agriculture, Penicillium species significantly impact plants, causing diseases and contamination that adversely affect crop yields and quality. Timely detection of Penicillium species is crucial for controlling disease and preventing mycotoxins from entering the food chain. To tackle this issue, we implement a novel species identification approach called Analysis of whole GEnome (AGE). Here, we initially applied bioinformatics analysis to construct specific target sequence libraries from the whole genomes of seven Penicillium species with significant economic impact: P. canescens, P. citrinum, P. oxalicum, P. polonicum, P. paneum, P. rubens, and P. roqueforti. We successfully identified seven Penicillium species using the target we screened combined with Sanger sequencing and CRISPR-Cas12a technologies. Notably, based on CRISPR-Cas12a technology, AGE can achieve rapid and accurate identification of genomic DNA samples at a concentration as low as 0.01 ng/µL within 30 min. This method features high sensitivity and portability, making it suitable for on-site detection. This robust molecular approach provides precise fungal species identification with broad implications for agricultural control, industrial production, clinical diagnostics, and food safety. Full article

Woodpeckers exhibit selectivity when choosing tree cavities for nest development in forest ecosystems, and fungi play a significant and important role in this ecological process. Therefore, there is a complex and intricate relationship between the various behaviors of woodpeckers and the occurrence of fungal species. Research into the complex bond between fungi and woodpeckers was undertaken to provide more information about this remarkable ecological relationship. Through the process of line transect sampling, woodpecker traces were searched for, and mist nets were set up to capture them. A total of 21 woodpeckers belonging to four species were captured. High-throughput sequencing of the ITS region was performed on fungal-conserved samples to enable an in-depth analysis of the fungal communities linked to the woodpeckers’ nests. Members of Ascomycota were the most abundant in the samples, accounting for 91.96% of the total, demonstrating the importance of this group in the forest ecosystem of this station. The statistical results indicate significant differences in the fungal diversity carried by woodpeckers among the different groups. Species of Cladosporium were found to be the most prevalent of all the detected fungal genera, accounting for 49.3%. The top 15 most abundant genera were Cladosporium, Trichoderma, Beauveria, Epicococcum, Hypoxylon, Penicillium, Nigrospora, Aspergillus, Oidiodendron, Cercospora, Talaromyces, Phialemo-nium, Petriella, Cordyceps, and Sistotrema. The standard Bray–Curtis statistical technique was used in a hierarchical clustering analysis to compute inter-sample distances, allowing for the identification of patterns and correlations within the dataset. We discovered that in the grouped samples from woodpeckers, there were differences in the diversity of fungal communities carried by four woodpecker species, but the less dominant fungal species were still similar. The findings highlight the need to consider these diverse ecological linkages in woodpecker research and conservation efforts. Full article

Palm kernel cake (PKC), a byproduct of palm oil extraction, serves an important role in Ecuador’s animal feed industry. The emergence of yellow-orange fungal growth in PKC on some cattle farms in Ecuador sparked concerns within the cattle industry regarding a potential mycotoxin-producing fungus on this substrate. Due to the limited availability of analytical chemistry techniques in Ecuador for mycotoxin detection, we chose to isolate and identify the fungus to determine its association with mycotoxin-producing genera. Through molecular identification via ITS region sequencing, we identified the yellow-orange fungus as the yeast Candida ethanolica. Furthermore, we isolated two other fungi—the yeast Pichia kudriavzevii, and the fungus Geotrichum candidum. Molecular identification confirmed that all three species are not classified as mycotoxin-producing fungi but in contrast, the literature indicates that all three have demonstrated antifungal activity against Aspergillus and Penicillium species, genera associated with mycotoxin production. This suggests their potential use in biocontrol to counter the colonization of harmful fungi. We discuss preventive measures against the fungal invasion of PKC and emphasize the importance of promptly identifying fungi on this substrate. Rapid recognition of mycotoxin-producing and pathogenic genera holds the promise of mitigating cattle intoxication and the dissemination of mycotoxins throughout the food chain. Full article

Soil and sediment host microorganisms are able to survive in extremely resource-limited environments. Therefore, more and more attention is being paid to cave sediments as a reservoir of microbiota. The aim of this study is the speleomycological evaluation of the culturable soil and sediment fungal communities in the Brestovská Cave. To explore the origins of fungi, speleomycological studies were conducted both inside and outside the cave under investigation. Additionally, two incubation temperatures (5 and 24 °C) were used to increase the species spectrum of isolated fungi. To achieve the most accurate species identification, we combined an assessment of morphological characteristics of the isolates with molecular sequencing (ITS, internal transcribed spacer). Twenty different species were found and the most frequent was Penicillium commune, followed by Trichosporiella cerebriformis and Pseudogymnoascus pannorum. To our knowledge, our study has enabled the first identification of fungal species such as Penicillium swiecicki, Cephalotrichum hinnuleum, Cosmpospora berkeleyana, Lecythophora hoffmannii, Ambomucor seriatoinflatus, and Mortierella minutissima in underground sites. Our data showed that the abundance and composition of the fungal community varied between the indoor and outdoor samples and thus from the entrance and less visited sites deeper in the cave. Full article