Westerdijk Fungal Biodiversity Institute

Prolonged cultivation of certain filamentous fungi, including Aspergillus terreus , on drug-free medium leads to degeneration and morphological heterogeneity, marked by the emergence of fluffy mycelium-type sectors. This phenomenon may indicate alterations in antifungal susceptibility profiles (particularly to amphotericin B (AmB) in A. terreus ), as well as reductions or losses in conidiation, sexuality, secondary metabolite production, and/or virulence. In the present study, various characteristics of an AmB-resistant wild-type (WT) strain and its AmB-susceptible sectorized derivative (ATSec) were characterized. Compared to WT, ATSec exhibited increased susceptibility to AmB, reduced sporulation, and comparable sterol contents and virulence in Galleria mellonella . To elucidate the genes involved in AmB resistance, gene expression levels were compared between WT and ATSec with and without AmB treatment. The expression of P-type ATPase-related genes, which are implicated in membrane composition changes and consequently in AmB resistance, was significantly higher in the WT strain compared to ATSec. Moreover, the up-regulation of genes involved in the biosynthesis of polyketides—a diverse group of secondary metabolites—was higher in WT compared to ATSec, with a significant number of these genes also carrying at least one mutation. The findings of this study indicate that P-type ATPases may significantly be involved in AmB susceptibility and resistance observed in ATSec and WT strains. Additionally, mutations in polyketide synthase genes in ATSec may contribute to the phenotypic alterations associated with the sectorized phenotype. IMPORTANCE Prolonged cultivation of certain filamentous fungi, including Aspergillus terreus, on drug-free medium leads to degeneration and morphological heterogeneity, marked by the emergence of fluffy mycelium-type sectors. This phenomenon may indicate alterations in antifungal susceptibility profiles (particularly to amphotericin B (AmB) in A. terreus), as well as reductions or losses in conidiation, sexuality, secondary metabolite production, and/or virulence. In the present study, various characteristics of an AmB-resistant wild-type strain (WT) and its AmB-susceptible sectorized derivative (ATSec) were characterized. Compared to WT, ATSec exhibited increased susceptibility to AmB, reduced sporulation, and comparable sterol contents and virulence in Galleria mellonella. To elucidate the genes involved in AmB resistance, gene expression levels were compared between WT and ATSec with and without AmB treatment. The expression of P-type ATPase-related genes, which are implicated in membrane composition changes and consequently in AmB resistance, was significantly higher in the WT strain compared to ATSec. Moreover, the up-regulation of genes involved in the biosynthesis of polyketides - a diverse group of secondary metabolites - was higher in WT compared to ATSec, with a significant number of these genes also carrying at least one mutation. The findings of this study indicate that P-type ATPases may significantly be involved in AmB susceptibility and resistance observed in ATSec and WT strains. Additionally, mutations in polyketide synthase genes in ATSec may contribute to the phenotypic alterations associated with the sectorized phenotype.

Background This systematic review and meta-analysis aimed to examine the performance of polymerase chain reaction (PCR) assays for diagnosing mucormycosis. Methods A standardised search was conducted from conception to December 3rd 2024 using PubMed, Embase, Global Health, and Cochrane library. Original studies that used PCR-based methods on any human specimen to diagnose mucormycosis were analysed for eligibility. Using a bivariate meta-analysis, the diagnostic performance of PCR was examined against the European Organisation for Research and Treatment of Cancer–Mycoses Study Group Education and Research Consortium 2020 (EORTC-MSGERC) definitions of proven and probable invasive mould disease, which was modified to include all patients at risk of mucormycosis. The study protocol was registered on the PROSPERO database (CRD42023478667). Findings Of 4855 articles, a total of 30 met inclusion criteria, including 5920 PCR reactions on 5147 non-duplicate specimens from 819 cases of proven/probable mucormycosis and 4266 patients who did not meet the EORTC- MSGERC 2020 criteria. According to specimen type, sensitivity of PCR varied (p < 0.001) whereas specificity was similar (p = 0.662). Bronchoalveolar lavage fluid offered the highest sensitivity of 97.5% (95% CI 83.7–99.7%), specificity of 95.8% (95% CI 89.6–98.4%), positive likelihood ratio (LR+) of 23.5, and negative likelihood ratio (LR−) of 0.03. Tissue provided sensitivity of 86.4% (95% CI 78.9–91.5%), specificity of 90.6% (95% CI 78.1–96.3%), LR+ of 9.2, and LR− of 0.15. Blood provided reduced sensitivity of 81.6% (95% CI 70.1–89.4%), specificity of 95.5% (95% CI 87.4–98.5%), DOR of 95, LR+ of 18.3, and LR− of 0.19. Formalin-fixed paraffin- embedded specimens yielded the lowest sensitivity of 73.0% (95% CI 61.0–82.3%), highest specificity of 96.4% (CI 95% 87.5–99.0%), LR+ of 20.2, and LR− of 0.28. The covariates best explaining heterogeneity of the overall analysis were specimen type, study design (cohort versus case-control) and disease prevalence while patient population (COVID-19 versus other) and PCR (conventional versus quantitative) had less impact on heterogeneity. Interpretation This meta-analysis confirms the high performance of PCR for diagnosing mucormycosis and supports the instatement of PCR detection of free-DNA in blood, BALF and tissue into future updated definitions and diagnostic guidelines for mucormycosis.

Fusarium root rot (FRR) of beans causes considerable losses in the production of common bean and cowpea, which are important staple foods in Brazil. Fusarium solani f. sp. phaseoli has been attributed as the etiological agent of this disease, but under the current understanding of the phylogenetic species concept, this forma specialis may represent different species. In this study, we propose to identify species of the Fusarium solani species complex – FSSC associated with FRR on common bean and cowpea based on molecular phylogenetic analyses and test its pathogenicity. A set of 66 strains was obtained from symptomatic plants collected in distinct geographical regions of Brazil. Based on the phylogenetic analyses of partial DNA sequences of RPB2, we identified F. paranaense (n = 44), F. solani sensu stricto (n = 8), F. suttonianum (n = 10), and F. martii (n = 2), all members of FSSC Clade 3. Two strains were identified as F. brasiliense, a member of Clade 2, according to analyses of RPB2, IGS and locus 44 regions. In pathogenicity tests on common bean and cowpea, all species induced similar symptoms, such as rotting from the main root extending to the hypocotyl and reduction of plant growth. To confirm that these fungi cause disease also in soybean, we inoculated soybean plants. There, F. brasiliense, F. paranaense, F. solani s. str., F. suttonianum, and F. martii induced root lesions. Fusarium brasiliense was the most virulent on all hosts in comparison with other FSSC species. With this study, we clarify the etiology of FRR in Brazil based on phylogenetic analysis and pathogenicity, complementing the concept of forma specialis. Additionally, we provide relevant support to disease management and plant breeding studies such as the selection of cultivars with resistance to Fusarium bean root rot in the field.

Xylooligosaccharides (XOS) are xylose oligomers produced from agro-industrial wastes through xylanases action. They show prebiotic properties that add value to commercial products, such as beverages and foods. Soy husk, rice bran, and corn cob were pretreated and hydrolyzed by recombinant xylanases with high catalytic activity, and the XOS were quantified. Finally, XOS were evaluated for their ability to selectively stimulate the growth of the probiotic Lactobacillus acidophilus. Three recombinant xylanases were produced, with emphasis on SM2 xylanase, which showed activity of 21,853.10 U/mL. Hydrolysis of corn cob generated the higher XOS concentrations, while rice bran saccharification led to high XOS diversity. Soybean husks showed a lower yield concerning XOS release, but it stood out in generating xylohexose. The in vitro prebiotic assay showed that L. acidophilus was able to grow and to ferment the produced XOS, generating mainly propionic acid, which was not observed for the pathogens. Graphical abstract

Fungal keratitis, which is caused primarily by Neocosmospora and Fusarium species, is a significant global health issue that affects more than a million people annually in tropical and subtropical regions. Neocosmospora solani (formerly Fusarium solani) is a leading cause of corneal infections, along with members of the Fusarium oxysporum species complex (FOSC). This study provides new insights by reporting a series of ocular fusariosis cases caused by FOSC members and presenting molecular evidence linking specific haplotypes within FOSC to human infections. We describe three cases of Fusarium keratitis selected from a comprehensive review of clinicopathological data in our institution's archives. These cases were chosen for their distinctive clinical presentations and the involvement of less common Fusarium species. Two of these patients were diagnosed with keratitis and anterior endophthalmitis, and the third patient had a corneal ulcer previously treated with topical antivirals and antibiotics. All patients were successfully treated with topical amphotericin B. The Fusarium isolates from these patients were subjected to detailed molecular characterization, including DNA sequencing (tef1α, rpb2, CaM, tub2, and LSU), amplified fragment length polymorphism (AFLP) marker analysis, and MALDI-TOF MS analysis. Remarkably, our study reports the first case of human infection by F. veterinarium, alongside cases involving F. contaminatum and F. curvatum. Furthermore, a molecular survey using haplotypic networks based on tef1α sequences identified genotypes associated with human infections and revealed the emergence of F. veterinarium clade VII. Our findings emphasize the need for vigilance regarding emerging species within the FOSC, particularly F. veterinarium. This highlights the need for improved diagnostic tools and targeted research to combat fusarioid-related infections effectively.

Aspergilli and other molds are prevalent in the environment and are an important cause of opportunistic infections and seasonal allergies in susceptible patients. This study determined species distribution of various molds in outdoor/indoor air in and around a major hospital and performed antifungal susceptibility testing and molecular fingerprinting of environmental and clinical Aspergillus fumigatus isolates in Kuwait. Sampling for the isolation of molds was performed for a 17-month-period from the water/indoor air of medical/surgical wards/ICUs and outdoor air. Molds were identified by phenotypic characteristics and/or by the PCR-sequencing of rDNA/β-tubulin/calmodulin genes. Antifungal susceptibility testing was done by Etest. Fingerprinting was performed by nine-loci-based microsatellite analysis. A total of 6179 isolates were obtained from outdoor (n = 4406) and indoor (n = 1773) environments. These included Cladosporium spp. (n = 2311), Aspergillus spp. (n = 1327), Penicillium spp. (n = 1325), Paecilomyces spp. (n = 473), Alternaria spp. (n = 218), Bipolaris spp. (n = 133), and other molds (n = 392). Fingerprinting data revealed heterogeneity among clinical and environmental A. fumigatus and shared genotypes among outdoor air and hospital environmental isolates. Itraconazole-resistant A. fumigatus isolates with TR34/L98H mutations in Cyp51A were also recovered from outdoor air (n = 1), a hospital environment (n = 3), and clinical samples (n = 2). More than 15 fungal genera and all four Aspergillus (Nigri, Flavi, Fumigati, and Terrei) sections and nine rare aspergilli were detected. The isolation frequency was higher during the peak allergy season of October/November. The presence of shared genotypes among outdoor air and the hospital environment including triazole-resistant A. fumigatus suggests a reservoir for invasive infections among susceptible hospitalized patients.

Aspergillus nidulans is an important model organism for eukaryotic biology and the reference for the section Nidulantes in comparative studies. In this study, we de novo sequenced the genomes of 25 species of this section. Whole-genome phylogeny of 34 Aspergillus species and Penicillium chrysogenum clarifies the position of clades inside section Nidulantes. Comparative genomics reveals a high genetic diversity between species with 684 up to 2433 unique protein families. Furthermore, we categorized 2118 secondary metabolite gene clusters (SMGC) into 603 families across Aspergilli, with at least 40 % of the families shared between Nidulantes species. Genetic dereplication of SMGC and subsequent synteny analysis provides evidence for horizontal gene transfer of a SMGC. Proteins that have been investigated in A. nidulans as well as its SMGC families are generally present in the section Nidulantes, supporting its role as model organism. The set of genes encoding plant biomass-related CAZymes is highly conserved in section Nidulantes, while there is remarkable diversity of organization of MAT-loci both within and between the different clades. This study provides a deeper understanding of the genomic conservation and diversity of this section and supports the position of A. nidulans as a reference species for cell biology.

Inflammatory diseases of the human gastrointestinal tract are affected by the microbes that reside in the mucosal surfaces. Patients with inflammatory bowel diseases (IBD) have altered bacterial and fungal intestinal compositions, including higher levels of fecal Candida yeasts. Ongoing research indicates that genetic and phenotypic diversity of Candida albicans may be linked with disease severity. Here, we set out to investigate feces-derived C. albicans strains from individuals with IBD and healthy volunteers through microsatellite-based genotyping and phenotypic assays. A seven-locus microsatellite panel was applied, of which six loci were newly developed. It appears that there is no specific lineage of C. albicans that is associated with IBD, but rather that the three study populations (Crohn's disease, ulcerative colitis, healthy volunteers) do have distinguishable distributions of genotypes. In addition, phenotypic characterization by means of enzyme release assays revealed trends between genotypes, virulence-related enzyme activity and clinical biomarkers. We thus show that microsatellite typing can describe genetic diversity of feces-derived C. albicans strains, and that phenotypic diversity of these strains may indeed correlate with fungal genotype or disease. This study opens further possibilities to investigate fecal fungi in relation to severity of inflammation in IBD or in other (intestinal) diseases.

The genus Aspergillus is diverse, including species of industrial importance, human pathogens, plant pests, and model organisms. Aspergillus includes species from sections Usti and Cavernicolus , which until recently were joined in section Usti , but have now been proposed to be non-monophyletic and were split by section Nidulantes , Aenei and Raperi . To learn more about these sections, we have sequenced the genomes of 13 Aspergillus species from section Cavernicolus ( A. cavernicola , A. californicus , and A. egyptiacus ), section Usti ( A. carlsbadensis , A. germanicus , A. granulosus , A. heterothallicus , A. insuetus , A. keveii , A. lucknowensis , A. pseudodeflectus and A. pseudoustus ), and section Nidulantes ( A. quadrilineatus , previously A. tetrazonus ). We compared these genomes with 16 additional species from Aspergillus to explore their genetic diversity, based on their genome content, repeat-induced point mutations (RIPs), transposable elements, carbohydrate-active enzyme (CAZyme) profile, growth on plant polysaccharides, and secondary metabolite gene clusters (SMGCs). All analyses support the split of section Usti and provide additional insights: Analyses of genes found only in single species show that these constitute genes which appear to be involved in adaptation to new carbon sources, regulation to fit new niches, and bioactive compounds for competitive advantages, suggesting that these support species differentiation in Aspergillus species. Sections Usti and Cavernicolus have mainly unique SMGCs. Section Usti contains very large and information-rich genomes, an expansion partially driven by CAZymes, as section Usti contains the most CAZyme-rich species seen in genus Aspergillus . Section Usti is clearly an underutilized source of plant biomass degraders and shows great potential as industrial enzyme producers.

The current paper represents the seventh contribution in the Genera of Fungi series, linking type species of fungal genera to their morphology and DNA sequence data. This manuscript focuses on a genus of dematiaceous hyphomycetes, Hirudinaria. Two species of this genus are treated in this study. Hirudinaria mespili, the type species of the genus, as well as Hirudinaria macrocarpa, are epitypified and provided with DNA sequence data to resolve their phylogeny as members of Mycosphaerellaceae (Mycosphaerellales, Dothideomycetes).

The aim of this study was to identify parameters influencing DNA extraction and PCR amplification efficiencies in an attempt to standardize Mucorales qPCR. The Fungal PCR Initiative Mucorales Laboratory Working Group distributed two panels of simulated samples to 26 laboratories: Panel A (six sera spiked with Mucorales DNA and one negative control serum) and Panel B (six Mucorales DNA extracts). Panel A underwent DNA extraction in each laboratory according to the local procedure and were sent to a central laboratory for testing using three different qPCR techniques: one in-house qPCR assay and two commercial assays (MucorGenius and Fungiplex). Panel B DNA extracts were PCR amplified in each laboratory using local procedures: nine in-house qPCR assays and two commercial kits (MucorGenius and MycoGENIE). All data were compiled and anonymously analyzed at the central laboratory. For Panel A, a total of six different automated platforms and five manual extraction methods were used. Positive rates were 64%, 70%, and 89%, for the MucorGenius, Fungiplex, and the in-house qPCR assay, respectively. Using a large volume of serum for DNA extraction provided the highest analytical sensitivity (82.5% for 1 mL compared with 62.7% for smaller volumes, P < 0.01). For Panel B, five in-house qPCR assays and two commercial kits had >78% positivity. Using larger PCR input volumes (≥7 µL) was associated with the highest sensitivity at 95.5% compared to 58.3% when lower input volumes were used (P < 0.01). Using larger sample volumes for nucleic acid extraction and DNA template volumes for PCR amplification significantly improves the performance of Mucorales qPCR when testing serum. IMPORTANCE Mucormycosis is a life-threatening mold infection affecting immunosuppressed patients but also other patients with diabetes or trauma. Better survival is linked to shorter delays in diagnosis and treatment initiation. Detection of Mucorales-free DNA in serum or plasma using quantitative PCR allows a prompt diagnosis and earlier treatment. Several techniques and protocols of quantitative Mucorales PCR are used in Europe, and improving performance remains a common objective of laboratories participating in the fungal PCR Initiative Working Group. This study, which combined results from 26 laboratories in Europe, showed that the main parameters underpinning sensitivity are the preanalytical variables (volume of serum used for DNA extraction and DNA template volume), irrespective of the extraction platforms and qPCR assay/platform.

Plant biomass degradation is of major importance for many fungi, as this is one of the most abundant and renewable carbon sources on earth. With a global push toward a bio-based economy based on plant biomass conversion, research in this area has obtained enormous momentum. In this chapter, recent insights obtained from fungal genomes and post-genomic studies related to plant biomass degradation will be discussed, focusing in particular on the diversity of the approaches used by different fungi. In addition, an updated overview will be presented of the enzymes currently considered to be involved in the degradation of the plant biomass polymers.

The exploration of new pharmacological compounds from endophytic fungi offers infinite possibilities. The aim of this study was to evaluate the antibacterial and antioxidant activities of extracts from the leaves of Ziziphus lotus and five of its endophytic fungi and investigate the chemical diversity of the secondary metabolites produced. Isolated, purified, and molecularly identified en-dophytes and plant leaves were subjected to ethyl acetate extraction. The antibacterial potential of the extracts was assessed by the disc diffusion method against five bacterial strains: Staphylococcus aureus ATCC 25923; Staphylococcus aureus MU50; Enterococcus faecalis WDCM00009; Escherichia coli ATCC 25922; and Pseudomonas aeruginosa ATCC 27853. DPPH and reducing power tests were performed to assess antioxidant potential. GC-MS analysis was used to identify volatile compounds in extracts. Fungal endophytes were identified as Aspergillus cavernicola, Aspergillus persii, Alternaria alternata, Cladosporium asperlatum, and Fusarium incarnatum-equiseti complex, with respective accession numbers DTO 412-G6, DTO 412-I5, DTO 413-E7, DTO 412-G4, and DTO 414-I2. GC-MS analysis revealed a large number of bioactive compounds. All extracts showed antibacterial activity against at least two of the bacteria tested, and most showed antioxidant activity. The Aspergillus cavernicola extract stood out for its higher phenolic content and higher antioxidant and antibacterial activities in all tests.

The species-rich Fusarium sambucinum species complex (FSAMSC; Fusarium , Nectriaceae , Hypocreales ) is well-known for including devastating plant pathogens and toxigenic species. However, this group of grass-loving fungi also accommodates soil saprobes, endophytes, mycoparasites and rare opportunistic pathogens of humans and other animals. Recent publications have highlighted the vast phylogenetic and biochemical diversity of the FSAMSC, although a large number of taxa in FSAMSC have not been systematically described and still lack Latin binomials. In this study we established the phylogenetic breadth of the FSAMSC using an integrative approach including morphological, multilocus phylogenetic, and coalescence analyses based on five gene regions (calmodulin, RNA polymerase II largest and second largest subunits, translation elongation factor 1-α, and β-tubulin). Results obtained support the recognition of 75 taxa in FSAMSC, including all the currently known species segregates of the Fusarium head-blight pathogen F. graminearum s. lat . Thirty novel species are formally described and illustrated, while four phylogenetic species remain undescribed. An epitype is proposed for the generic type of Fusarium , F. sambucinum , from recently collected material identified by means of morphology, phylogenetics and mating experiments, fixing the phylogenetic application of the name. Additional notes are included on the typification of Fusisporium cerealis (syn. Fusarium cerealis ).

Infectious keratitis is a significant ocular disease that, if left untreated, can lead to blindness. Fungi are among the causative agents that can result in severe symptoms. Keratitis infections are prevalent globally, with a higher incidence reported in tropical and subtropical regions. The current research focused on the molecular diagnosis of fungal keratitis and its prevalence over a 3-year period in northeastern Iran. The study involved the collection of 38 corneal scraping specimens from the Eye Specialized Hospital of Khatam in Mashhad, northeastern Iran. These specimens were cultured on Sabouraud dextrose agar, and the isolates were identified using DNA-based techniques. Among the patients studied ( n = 38), 22 (58%) cases were caused by Aspergillus species ( A. flavus , n = 17, A. fumigatus , n = 3; A. terreus , n = 1; A. tubingensis , n = 1), seven (18%) by Neocosmospora species ( N. falciformis , n = 4; N. solani , n = 3), three (7%) by Candida albicans , two (5%) by Fusarium annulatum , and one case each (2%) by Penicillium chrysogenum , Cladosporium cladosporioides , and Cytospora sp. In addition, one case had a combined infection of A. flavus and P. glabrum . The results indicate a higher incidence of fungal keratitis in males, particularly in the age range of 40–60 years. Aspergillus sp., and specifically A. flavus , had the highest prevalence. Cladosporium cladosporioides is reported for the first time in this area as causal agent of keratitis. Additionally, this is the first report of keratitis likely caused by Cytospora species.

The Mucorales is a group of ancient fungi with global distribution. In the current study we accessed mucoralean fungi isolated from two countries on opposite sides of the Earth and in different hemispheres: South Korea and Brazil. Mucorales isolates were obtained from freshwater, soil, invertebrates, and fruit seeds and identified using phenotypic techniques combined with the DNA sequence data. These analyses revealed 15 new species including one that we affiliated to a newly proposed genus, Neofenellomyces . Names proposed for these 15 new species are Absidia cheongyangensis , A. fluvii , A. kunryangriensis , A. paracylindrospora , A. tarda , A. variiprojecta , A. variispora , Backusella varians , Mucor albicolonia , M. aurantiacus , M. cryophilus , M. glutinatus , M. paraorantomantidis , M. timomeni , and Neofennellomyces jeongsukae . Of these new species, 12 were isolated from South Korea: A. cheongyangensis , A. fluvii , A. kunryangriensis , A. paracylindrospora , B. varians , M. albicolonia , M. aurantiacus , M. cryophilus , M. glutinatus , M. paraorantomantidis , M. timomeni , and N. jeongsukae , and three from Brazil: A. tarda , A. variiprojecta , and A. variispora . Niche specificity of these fungi is discussed including newly recorded invertebrate hosts and a new geographic distribution for species of Backusella , Circinella , Cunninghamella , and Mucor . Given these findings, we provide an inventory of Mucorales .

Cytospora species have commonly been reported as important plant pathogenic fungi with wide host ranges and geographic distributions. With the increase in the number of cryptic species being described, a comprehensive global taxonomic revision of the genus Cytospora is required. The present study includes 399 isolates from 32 countries. These isolates were subjected to DNA sequence analysis for five genomic loci (ITS, act1 , rpb2 , tef1-α and tub2 ). Based on these data, it could be confirmed that Cytospora , Leucostoma , Valsa , Valsella and Valseutypella are congeneric. Furthermore, 111 species of Cytospora could also be reassessed, 44 species and four combinations newly introduced, and new typifications proposed for a further three species. Three asexual morphological groups (including 13 asexual morphological types) and three sexual morphological groups (including eight sexual morphological types) were designated. The present study explored the species diversity of Cytospora and re-evaluated the identity of all cultures in the Westerdijk Fungal Biodiversity Institute (Utrecht, The Netherlands) that were deposited as either Cytospora or as one of its related genera. This is the most comprehensive phylogenetic analysis thus far conducted on Cytospora and the results contribute to an increased understanding of the taxonomy of these important fungi. It is also hoped that the findings will lead to improved management strategies for diseases associated Cytospora species.

During entomopathogenic fungal surveys conducted in Thailand, 15 specimens tentatively classified under Akanthomyces sensu lato were identified. To gain a comprehensive understanding of their taxonomy, molecular phylogenies using combined LSU, TEF1, RPB1, and RPB2 sequence data, together with morphological examination of several Akanthomyces spp. from previous studies were conducted. The analyses revealed distinct clades representing independent lineages within the Cordycipitaceae . These clades were further characterized by different asexual morph types and the respective hosts they parasitize. In this context, we resurrected the genus Lecanicillium to accommodate 12 known species previously classified under Akanthomyces sensu lato , found on diverse hosts. We propose four new genera ? Corniculantispora, Corpulentispora, Zarea, and Zouia ? from species previously identified as Lecanicillium . Notably, certain Akanthomyces species associated with spiders and parasitic on Ophiocordyceps sinensis were reclassified into the new genera Arachnidicola and Kanoksria , respectively. Moreover, we introduce four novel species in Akanthomyces sensu stricto found across a diverse range of moth families: Ak. buriramensis, Ak. fusiformis, Ak. niveus, and Ak. phariformis. Additionally, we provide descriptions and illustrations of the sexual morph linked to Ak. laosensis and Ak. pseudonoctuidarum, along with a second type of synnemata seen in Ak. noctuidarum and Ak. pseudonoctuidarum. To assist with their identification, keys to the genera Akanthomyces, Arachnidicola , and Lecanicillium are provided, but should not be used to replace molecular identification.