Culturable fungi from urban soils in China II, with the description of 18 novel species in Ascomycota (Dothideomycetes, Eurotiomycetes, Leotiomycetes and Sordariomycetes).

As China's urbanisation continues to advance, more people are choosing to live in cities. However, this trend has a significant impact on the natural ecosystem. For instance, the accumulation of keratin-rich substrates in urban habitats has led to an increase in keratinophilic microbes. Despite this, there is still a limited amount of research on the prevalence of keratinophilic fungi in urban areas. Fortunately, our group has conducted in-depth investigations into this topic since 2015. Through our research, we have discovered a significant amount of keratinophilic fungi in soil samples collected from various urban areas in China. In this study, we have identified and characterised 18 new species through the integration of morphological and phylogenetic analyses. These findings reveal the presence of numerous unexplored fungal taxa in urban habitats, emphasising the need for further taxonomic research in urban China.

Keratinophyton chongqingense sp. nov. and Keratinophyton sichuanense sp. nov., from soil in China.

Keratinophyton is a genus of well-known keratinophilic fungi found in various terrestrial habitats. During a survey of keratinolytic fungi in China, a total of 12 isolates of Keratinophyton species, representing eight taxa, were obtained from the soil. Two of these isolates were described as new species based on their morphological characteristics and molecular analyses of the internal transcribed spacer region and the rRNA gene of the nuclear large subunit. Descriptions and illustrations of these two novel species, which are named Keratinophyton chongqingense sp. nov. and Keratinophyton sichuanense sp. nov., are provided herein.

Survey of Keratinophilic Fungi from Feathers of Birds in Tuscany.

Although keratinophilic fungi on avian feathers have been widely described, data from European literature are quite lacking regarding Mediterranean countries. The aim of the present study was therefore to evaluate the occurrence of fungal species on feathers of different avian species in Italy. A total of 378 feather samples from both aquatic (n = 254) and terrestrial birds (n = 124), for a total of 30 bird species, were cultured for keratinophilic fungi. Fungal isolates were recognized by their macro- and micro-scopical morphology, and results were corroborated by PCR and sequencing. Keratinophilic fungi belonging to 11 different species (Scopulariopsis brevicaulis, Chrysosporium keratinophilum, Trichophyton terrestre, Microsporum gypseum, Sepedonium sp., Chrysosporium pannorum, Myriodontium sp., Chrysosporium tropicum, Chrysosporium pruinosum, Chrysosporium luteum and Aphanoascus fulvescens) were isolated from 71 animals (18.8%). The frequency of isolation of keratinophilic species from terrestrial birds was significantly higher in waterfowl. Migratory birds in Italy have been proven to carry pathogenic fungi such as dermatophytes, (A. platyrhyncos, A. crecca, E. rubecula), besides saprophytic species.

Culturable Fungi from Urban Soils in China I: Description of 10 New Taxa.

An investigation of members of the soil keratinophilic fungi community in China resulted in the identification of one new monotypic genus, Zongqia, and 10 new species, 2 of which are affiliated with Solomyces, 1 with the new genus Zongqia, 4 with Pseudogymnoascus, and 3 with Scedosporium. These novel taxa form an independent lineage distinct from other species, based on morphological and multilocus phylogenetic analyses. Descriptions, illustrations, and notes are provided for each taxon. These new taxa of the soil keratinophilic fungi add to the increasing number of fungi known from China, and it is now evident that numerous novel taxa are waiting to be described. IMPORTANCE Keratinophilic fungi are a group that can degrade and utilize keratin-rich material. It is also because of this ability that many taxa can cause infections in animals or humans but remain poorly studied. In this study, we reported a novel genus and 10 novel species, 7 novel species belonging to the order Thelebolales and 3 to the genus Scedosporium, based on multilocus phylogenetic analyses combined with morphological characteristics. Our study significantly updates the taxonomy of Thelebolales and Scedosporium and enhances our understanding of this group of the keratin-degrading fungal community. The findings also encourage future studies on the artificially constructed keratin-degrading microbial consortia.

Molecular systematics of Keratinophyton: the inclusion of species formerly referred to Chrysosporium and description of four new species.

Four new Keratinophyton species (Ascomycota, Pezizomycotina, Onygenales), K. gollerae, K. lemmensii, K. straussii, and K. wagneri, isolated from soil samples originating from Europe (Austria, Italy, and Slovakia) are described and illustrated. The new taxa are well supported by phylogenetic analysis of the internal transcribed spacer region (ITS) region, the combined data analysis of ITS and the nuclear large subunit (LSU) rDNA, and their phenotype. Based on ITS phylogeny, within the Keratinophyton clade, K. lemmensii is clustered with K. durum, K. hubeiense, K. submersum, and K. siglerae, while K. gollerae, K. straussii and K. wagneri are resolved in a separate terminal cluster. All four new species can be well distinguished from other species in the genus based on phenotype characteristics alone. Ten new combinations are proposed for Chrysosporium species which are resolved in the monophyletic Keratinophyton clade. A new key to the recognized species is provided herein.

Keratinophilic fungi from the feathers of Emu (Dromaius novaehollandiae) in Maharastra India.

A total of 70 feathers samples of Emu (Dromaius novaehollandiae) were collected from 7 Emu farms situated at two districts (Raigad and Thane) of Maharashtra (India) and screened for resident keratinophilic fungi. Among them, 44 isolates were recovered and identified by evaluating characteristic macro- and micro-morphological features. Further gene products corresponding to the ITS1-5.8S-ITS2 rDNA region from all isolates were amplified and sequenced. Homology search was performed using BLAST program against non-redundant nucleotide database, and significantly matched DNA sequences deposited to the NCBI Gene Bank for reference purposes. Eight identified fungal species belongs to 7 different genera named as Aphanoascus terreus Ac_MW577456 (21.43%), Microsporum gypseum Ac_MW580920 (14.29%), Ctenomyces serratus Ac_MW577459 (10.0%), Uncinocarpus orissi Ac_MW577461 (5.17%), Aphanoascus verrucosus Ac_MW577458 (4.29%), Gymnascella dankaliensis Ac_MW577460 (2.86%), Gymnoascoideus petalosporus Ac_MW577462 (2.86%) and Arthroderma tuberculatum Ac_MW577457 (1.43%).

Three-gene phylogeny of the genus Arthroderma: Basis for future taxonomic studies.

Arthroderma is the most diverse genus among dermatophytes encompassing species occurring in soil, caves, animal burrows, clinical material and other environments. In this study, we collected ex-type, reference and authentic strains of all currently accepted Arthroderma species and generated sequences of three highly variable loci (ITS rDNA, ß-tubulin, and translation elongation factor 1-α). The number of accepted species was expanded to 27. One novel species, A. melbournense (ex-type strain CCF 6162T = CBS 145858T), is described. This species was isolated from toenail dust collected by a podiatrist in Melbourne, during an epidemiological study of four geographical regions of Eastern Australia. Trichophyton terrestre, Chrysosporium magnisporum, and Chrysosporium oceanitis are transferred to Arthroderma. Typification is provided for T. terrestre that is not conspecific with any of the supposed biological species from the former T. terrestre complex, that is, A. insingulare, A. lenticulare and A. quadrifidum. A multi-gene phylogeny and reference sequences provided in this study should serve as a basis for future phylogenetic studies and facilitate species identification in practice. LAY ABSTRACT: The genus Arthroderma encompasses geophilic dermatophyte species that infrequently cause human and animal superficial infections. Reference sequences from three genetic loci were generated for all currently accepted Arthroderma species and phylogeny was constructed. Several taxonomic novelties are introduced. The newly provided data will facilitate species identification and future taxonomic studies.

Diversity of Onygenalean Fungi in Keratin-Rich Habitats of Maharashtra (India) and Description of Three Novel Taxa.

Extensive survey was carried out in the state of Maharashtra, India, as part of a 3-year project to explore keratinophilic fungal diversity for conservation and biotechnological potential. A total of 578 soil samples were collected from keratin-rich habitats across 24 districts of Maharashtra State. Hair-baiting technique and micro-dilution drop-trail method were employed for isolation and purification of keratinophilic fungi from soil. A total of 66 species belonging to 17 genera of order Onygenales were recorded in hair baits. Eleven taxa were found to be new to science, most of which were rare as they were recorded in only one sample out of the > 500 samples analyzed. Three novel taxa have been characterized at morphological and molecular level and described here as new to science. These taxa include Currahmyces indicus gen. et sp. nov., Canomyces reticulatus gen. et sp. nov., Ctenomyces indicus sp. nov. All these novel taxa are morphologically and phylogenetically distinct from known taxa of order Onygenales. The study indicates that systematic sampling of a larger area is needed to uncover the hidden (unknown) diversity of keratinophilic fungi which is overlooked in sporadic samplings as evident from previous studies.

DNA sequence analysis of keratinophilic fungi isolated from livestock stables in the East of Iran.

BACKGROUND AND PURPOSE: Keratinophilic fungi play an important role in the decomposition of keratinous substances in nature. This capacity induces dermatomycosis in both humans and livestock. The soil of livestock stables can be a reservoir of keratinophilic fungi. Therefore, the present study was conducted to isolate and identify keratinophilic fungi in the soil of the livestock stables located in Qayen, South Khorasan Province, Iran. MATERIALS AND METHODS: This study was conducted on 62 soil samples collected from livestock stables. The samples were cultured by means of hair bait technique (HBT). The identification of the isolates was performed based on their morphological characteristics and then confirmed by polymerase chain reaction and sequencing of the ITS regions of ribosomal DNA. RESULTS: A total of 118 isolates of 7 species from 5 genera were identified. Aphanoascus verrucosu (n=70, 59.36%) was detected as the dominant keratinophilic fungus, followed by Arthroderma quadrifidum (n=13, 11.01%), A. terreus (n=12, 10.16%), Acremonium (n=12, 10.16%), A. gertleri (n=5, 4.23%), Fusarium equiseti (n=3, 2.54%), and Uncinocarpus reesii (n=3, 2.54%). CONCLUSION: Different keratinophilic fungi were isolated from the soil of livestock stables; however, A. verrucosu was found to be the dominant species.

Keratinophilic fungi from the vicinity of salt pan soils of Sambhar lake Rajasthan (India).

Forty soil samples were collected from seven sites in the vicinity of Sambhar lake Rajasthan, India and screened for the presence of keratinophilic fungi using hair baiting techniques for isolation. Seventeen isolates were recovered and identified. The cultures were identified by recognition of their macro- and micro- morphological features. Their identification was confirmed by BLAST using ITS1-5.8S-ITS2 rDNA region and sequences have been deposited in NCBI data base. A total of 34 species belonging to 29 genera were isolated. Among the dermatophytes and related keratinophilic fungi Chrysosporium indicum was predominant followed by Ctenomyces serratus, C. tropicum, Keratinophyton durus, Auxarthron conjugatum, Gymnascella dankaliensis, Gymnoascoideus petalosporus and Uncinocarpus reesii. Twenty-six species belonging to 22 genera represented other species. Our study indicates that keratinophilic fungi and species are found in the soils at the vicinity of the Sambhar Lake, and human activities can be the potential source of pathogenic fungi.

Incidence of Keratinophilic Fungi from the Selected Soils of Kaziranga National Park, Assam (India).

Seventy-eight soil samples were collected from the various locations in the vicinity of Kaziranga National Park (Assam), India, during April to October 2009 and screened for the presence of keratinophilic fungi using the hair baiting techniques for isolation. Thirty-nine isolates were recovered and identified by recognition of their macro- and micromorphological features. Their identification was also confirmed by the BLAST search of sequences of the ITS1-5.8S-ITS2 rDNA region against the NCBI/GenBank data and compared with deposited sequences for identification purpose. Eleven species related to seven genera were recorded viz. Aphanoascus durus (1.28%), Arthroderma tuberculatum (3.84%), Arthroderma corniculatum (1.28%), Chrysosporium indicum (16.66%), C. tropicum (3.84%), Ctenomyces serratus (5.12%), Keratinophyton punsolae (1.28%), Microsporum appendiculatum (1.28%), Microsporum gypseum complex (11.53%), Trichophyton mentagrophytes (11.28%) and T. terrestre (2.56%).

Isolation of keratinophilic fungi from the soil of Greater Tunb, Abu-Musa, and Sirri islands in the Persian Gulf, Iran.

BACKGROUND AND PURPOSE: Keratinophilic fungi are among the important groups of fungi living in the soil. This study aimed to isolate and identify keratinophilic fungi from the soil of three Iranian islands, namely Greater Tunb, Abu Musa, and Sirri, located in the Persian Gulf using morphological and molecular (polymerase chain reaction) methods. MATERIALS AND METHODS: In this study, a total of 60 soil samples were collected from the three islands of Greater Tunb, Abu Musa, and Sirri. The samples were analyzed for the presence of the keratinophilic fungi using a hair baiting technique. Furthermore, the identification of keratinophilic fungi was accomplished through the employment of molecular and sequencing techniques. RESULTS: A total of 130 fungal isolates, including 11 genera with 24 species, were collected. Accordingly, Chrysosporium tropicum (24;18.5%), C. keratinophilum (17; 13.1%), Chrysosporium species (15; 11.5%), Aspergillus species ( 8;6.1%), Aspergillus flavus (8; 6.1%), Penicillium species (8;6.1%), Alternaria spp ( 6; 4.6%), Phoma species (5; 3.8%), Aphanoascus verrucosus (4;3.1%), Fusarium chlamydosporum (4; 3.1%), Aspergillustrreus (4;3.1%), Acremonium species (4; 3.1%), and other fungi( 23; 17.8 %) isolates were identified . All isolates of keratinophilic fungi were isolated from the soils with the pH range of 7-9. CONCLUSION: The results of this study contributed towards a better conceptualization of the incidence pattern of keratinophilic fungi in the regions of Iran. Given that no study has investigated this issue, the findings of the present study can be beneficial for the management of public health surveillance, physicians, and epidemiologists.

Preferential utilization and colonization of keratin baits by different myco-keratinophiles.

Myco-keratinophilic species have a predilection for different keratinous substrates but show variability in their affinity towards them. Keeping this in view, a survey was conducted in the Khardung and Khardung La soils of Ladakh (India) and 28 myco-keratinophilic species belonging to 15 fungal genera (Sarocladium, Aspergillus, Beauveria, Chrysosporium, Cladosporium, Alternaria, Epicoccum, Fusarium, Gibberella, Clonostachys, Paecilomyces, Purpureocillium, Metarhizium, Penicillium and Sagenomella) were isolated by using keratin bait technique. These isolated species were tested for their preferential utilization ability and colonization on different baits by morphological assessment. Different types of keratin baits used were feathers, human hair, human nails and wool. Overall assessment revealed that feathers were colonized and utilized by all the species (100 %), followed in decreasing order by nails (89.29 %), hair (85.71 %) and sheep wool (67.86 %). So, it is concluded that feather baiting technique, could be more useful in trapping keratinophilic fungi than the hair baiting technique which is till date regarded as the best method for the isolation of myco-keratinophiles. On the basis of succession on keratinous baits, the recovered keratinophilic species were also categorized into four categories: early successional species (pioneer colonizers), late successional species (final colonizers), persistent species and no-pattern species.

[Keratinophilic fungi in soils of parks of Corrientes city, Argentina]. / Hongos queratinofílicos en suelos de parques de la ciudad de Corrientes, Argentina.

BACKGROUND: The soil is a natural reservoir of keratinophilic fungi, which are a small but important group of filamentous fungi, some of which typically develop on keratinized tissues of living animals. There are numerous species of saprophytic fungi with recognized keratinophilic abilities, and several studies have been undertaken in order to link their presence to possible human disease. AIMS: To know the biota of geophilic fungi in general and of keratinophilic fungi particularly in soils from two public parks. METHODS: Soil samples from two public parks of Corrientes city, Argentina, were studied during two seasons, using the hook technique and serial dilutions for fungal isolation. RESULTS: Using the hook technique, 170 isolates were classified into 17 genera and 21 species, among which it is worth mentioning the presence of Microsporum canis. Shannon index for keratinophilic fungi in autumn was 2.27, and 1.92 in spring. By means of the serial dilutions technique, 278 fungi isolated were identified into 33 genera and 71 species. Shannon index in autumn was 3.9, and 3.5 in spring. CONCLUSIONS: The soils studied have particularly favorable conditions for the survival of pathogens and opportunistic geophilic fungi for humans and animals.

Mycological and Molecular Analysis of Soil Keratinophilic Fungi Isolated in Summer Beaches of the Korean South Coast / 대한피부과학회지

BACKGROUND: Some species of soil keratinophilic fungi (KPF) are known to be pathogens that may lead to cutaneous infection. People exposed to these species through direct contact with soil on beaches can contract KPF infection. However, there is little literature regarding pathogenic KPF isolated from beaches during summer time. OBJECTIVE: This study aims to evaluate the distribution of soil KPF isolated from beaches in Korea during summer. METHODS: One hundred eighty soil samples from six beaches in the southern coastline of Korea under three different climatic conditions were collected. The KPF species were isolated using the hair-baiting technique. Then, molecular identification was performed by sequencing the ribosomal RNA (rRNA) internal transcribed spacer (ITS) region to investigate the exact species of the isolated fungi. RESULTS: Among the one hundred eighty soil samples, twenty-nine strains (16.1%) of KPF were recovered. The isolation rate of KPF among the beaches varied from 0 to 34.5%. KPF was most frequently isolated in shaded dry areas (30%), followed by sunny dry areas (18.3%), and sunny wet areas (0%). Molecular identification of the fungi using rRNA ITS analysis helped in their classification. Microsporum gypseum/Arthroderma incurvatum (69.0%), Microsporum gypseum/Arthroderma gypseum (3.4%), Trichophyton ajelloi/Arthroderma uncinatum (13.8%), Microsporum cookei/Arthroderma cajetani (10.3%), and Chrysosporium indicum/Aphanoascus terreus (3.4%) were identified. Single nucleotide polymorphism (SNP) was observed at position 180 of the rRNA ITS2 in the 20 strains of Microsporum gypseum/Arthroderma incurvatum, and the species was divided into Types 1 (14 strains) and 2 (6 strains) depending on the base present at the SNP position. The geographic distribution of these two types differed. CONCLUSION: Our results show that the beach is a possible source of keratinophilic fungal infection in humans. People should be aware of pathogenic fungi on the soil of beaches during summer and take measures to prevent possible superficial fungal infections.

Isolation Rates of Fungi from Soil Near Korean Wrestling Fields, Playgrounds, and Beauty Salons in Daegu / 대한피부과학회지

BACKGROUND: Soil is one of the major habitats of fungi. Keratinophilic fungi are an important group of fungi that live in soil, some of which are potential pathogens for humans and animals. OBJECTIVE: The aim of this study was to isolate and identify keratinophilic fungi near Korean wrestling fields, playgrounds, and beauty salons. METHODS: We visited 5 Korean wrestling fields, 15 playgrounds, and 10 beauty salons in Daegu, Korea, from July to August 2015. Soil samples were divided into 3 groups according to the distance from Korean wrestling fields, playgrounds, and beauty salons: near, 10 m, and 100 m. RESULTS: Of 185 soil samples examined, 60 (32.4%) yielded keratinophilic fungi. Specifically, of the 125 samples around Korean wrestling fields and playgrounds, isolation rates were 37.7% (17/45) at the near distance, 27.5% (11/40) at 10 m, and 0% at 100 m. Of the 60 samples around beauty salons, isolation rates were 85% (17/20) at the near distance, 75% (15/20) at 10 m, and 0% at 100 m. Near Korean wrestling fields, playgrounds, and beauty salons, Penicillium spp. were the predominant species (16.9%, 11/65), followed by Fusarium spp. (13.8%, 9/65), Purpureocillium spp. (12.3%, 8/65), Microsporum spp. (6.1%, 4/65), Chrysosporium spp. (3.1%, 2/65), and other species. CONCLUSION: In this study, isolation rates of keratinophilic fungi were higher in the soil closer to the Korean wrestling fields, playgrounds, and beauty salons. Especially near beauty salons, we found keratinophilic fungi in most soil samples. We should be aware of these results and pay attention to eliminate hair or keratins in soils.

Discovery of a sexual stage in Trichophyton onychocola, a presumed geophilic dermatophyte isolated from toenails of patients with a history of T. rubrum onychomycosis.

Trichophyton onychocola is a recently described geophilic dermatophyte that has been isolated from a toenail of Czech patient with a history of onychomycosis due to T. rubrum and clinical suspicion of relapse. In this study, we report a similar case from Denmark in an otherwise healthy 56-year-old man. The patient had a history of great toenail infection caused by T. rubrum in 2004 and presented with suspected relapse in 2011 and 2013. Trichophyton onychocola was the only microbial agent isolated at the second visit in 2013 and the identification was confirmed by DNA sequencing. Direct microscopic nail examination was positive for hyphae, however the etiological significance of T. onychocola was not supported by repeated isolation of the fungus. This new species may be an overlooked geophilic species due to the resemblance to some common species, for example, zoophilic T. interdigitale or some species of geophilic dermatophytes. We included differential diagnosis with phenotypically similar species; however, it is recommended that molecular methods are used for correct identification. The MAT locus of Danish strain was of opposite mating type than in the previously isolated Czech strain and the two isolates were successfully mated. The mating experiments with related heterothallic species T. thuringiense and Arthroderma melis were negative. The sexual state showed all typical signs of arthroderma-morph and is described by using optical as well as scanning electron microscopy. The sexual state was induced on a set of agar media, however low cultivation temperature and the presence of keratin source were crucial for the success rather than formulation of medium.

Isolation of keratinophilic fungi from selected soils of Sanjay Gandhi National Park, Mumbai (India).

One hundred and twenty-five samples were collected from eight different sites in the vicinity of Sanjay Gandhi National Park (SGNP) and screened for the presence of keratinophilic fungi using hair baiting technique for isolation. Seventy-three isolates were recovered and identified. The cultures were identified using macro- and micro-morphological features. Their identification was also confirmed by the BLAST search of sequences of the ITS1-5.8S-ITS2 rDNA region against the NCBI/Genbank data and compared with deposited sequences for identification purpose. Thirteen species of nine genera were isolated viz. Aphanoascus durus (2.4%), Arthroderma corniculatum (1.6%), Auxarthron umbrinum (0.8%), Chrysosporium evolceanui (1.6%), Chrysosporium indicum (16.0%), Chrysosporium tropicum (2.4%), Chrysosporium zonatum (4.0%), Chrysosporium states of Arthroderma tuberculatum (0.8%), Chrysosporium state of Ctenomyces serratus (11.2%), Gymnascella dankaliensis (3.2%), Microsporum gypseum (12.0%), Myriodontium keratinophilum (0.8%) and Trichophyton mentagrophytes (1.6%). Representative of all thirteen species can release the protein in the range of 152.2-322.4 µg/mL in liquid media when grown on human hair in shake flask culture and also decompose 18.4-40.2% of human hair after four weeks of incubation. This study indicates that the soils of SGNP, Mumbai may be significant reservoirs of certain keratinophilic fungi. The keratinolytic activity of these fungi may be playing significant role in superficial infections to man and animals and recycling of keratinic material of this environment.

A preliminary suudy on the occurrence of keratinophilic fungi in soils of Jamaica / Estudo preliminar sobre a ocorrência de fungos queratinofílicos em solos da Jamaica

This report represents the first study of keratinophilic fungi present in soils of Jamaica. Out of the 40 soil samples examined from different habitats, 30 (75%) were positive for the presence of keratinophilic fungi, yielding 36 isolates of keratinophilic fungi. Microsporum gypseum complex (represented by 16 isolates of M. gypseum, and four of M. fulvum) was most frequent, being present in 50% of the samples. A very high occurrence of this dermatophyte in Jamaican soil is of public health significance. The remaining isolates of keratinophilic fungi were represented by Chrysosporium spp (mainly C. indicum and C. tropicum) and Sepedonium sp.

Esta comunicação representa o primeiro estudo sobre fungos queratinofílicos presentes em solos da Jamaica. De 40 amostras de solo examinadas de diferentes localidades, 30 (75%) foram positivas para a presença de fungos queratinofílicos permitindo 36 isolamentos dos mesmos. O complexo Microsporum gypseum (representados por 16 isolamentos de M. gypseum e quatro de M. fulvum) foi o mais frequente, estando presente em 50% das amostras. A muito alta ocorrência deste dermatófito no solo da Jamaica é significante para a saúde pública. Os isolados remanescentes de fungos queratinofílicos foram representados pelo Chrysosporium spp (principalmente C. indicum e C. tropicum) e Sepedonium sp.