Analysis Of ITS Sequences Research Articles

Comparison of Endophytic and Epiphytic Microbial Communities in Surviving and Dead Korean Fir (Abies koreana) Using Metagenomic Sequencing

Plant endophyte and epiphyte communities cooperatively interact with their host plants and play crucial roles in sustaining plant fitness. In Korea, a variety of studies have been conducted to elucidate the reasons for the declining population of the endangered Korean fir (Abies koreana), but the relationship between microbiota and the healthy condition of trees remains unclear. Here, we conducted bacterial 16S rRNA gene and fungal ITS sequence analyses to dissect the composition of endophytic and epiphytic microbiota in both live and dead trees located in the same Mt. Jiri habitat. In the live trees, the bacterial class Armatimonadia and the lichenized fungi groups were significantly dominant, whereas many bacterial and fungal taxa mainly found in rotten wood were enriched in the dead trees. Functional prediction of the microbial communities in live trees suggested the possibility that bacterial endophytes and epiphytes play a role in inorganic nutrient metabolism and fungal endophytes and epiphytes produce biologically active secondary metabolites, thereby contributing to the healthy condition of Korean fir trees. The ecological function of endophytes and epiphytes in dead trees was predicted to be involved in the decomposition of wood for nutrient recycling. Our analyses revealed a distinct difference in microbial communities depending on the health condition of Korean fir trees. The results from this study would be useful for understanding the ecological function of endophytic and epiphytic microorganisms to conserve and manage this endangered species from ecologically vulnerable environments.

New insight into morphological and genetic diversity of Phlyctema vagabunda and Neofabraea kienholzii causing bull's eye rot on apple and pear

AbstractFungi of genera Phlyctema and Neofabraea are the causal agents of bull's eye rot, a major postharvest disease of pome fruits. To investigate their morphological and genetic diversity, isolates obtained in Italy and Chile from decayed fruit and rainwater between 2014 and 2019 were grown on two agar media, inoculated onto four fruit cultivars and compared using four marker genes. Consistent intra‐ and interspecies phenotypic differences were recorded among isolates identified as P. vagabunda (two main morphotypes, PvM‐I and PvM‐II, were distinguished) and N. kienholzii. In particular, the Chilean isolates belonging to PvM‐I showed low sporulation in vitro, while isolates belonging to PvM‐II showed the most abundant sporulation and also formed conidiomata deep within fruit tissue. Host cultivar influenced the disease incidence in unwounded, inoculated fruit. Cripps Pink and Golden Delicious apples favoured the formation of P. vagabunda conidiomata and macroconidia, while Granny Smith apples and/or Kaiser pears restricted sporulation of some isolates of PvM‐I. Mycelial cords of P. vagabunda and N. kienholzii were consistently recorded in inoculated fruit, suggesting their possible involvement as a source of inoculum. Propagules of P. vagabunda were present in rainwater collected from apple plants from September to October in Italy. According to sequence analysis of ITS, EF‐1α, TUB2 and ACT1 regions of the fungi, 12 distinct sequence types were identified, three of which were characteristic of isolates from the Southern Hemisphere. The condensed maximum‐likelihood phylogenetic tree separated the 50 P. vagabunda isolates into six phylogroups, suggesting a correlation with their geographical distribution.

A new species of Agaricus sect. Agaricus (Agaricaceae) from India

Agaricus albovariabilis is presented as a new species from peninsular India on the basis of morphological features and ITS sequence analysis. It is charcterized by small to medium sized white basidiomata, becoming brownish at maturity, unchanging context, mild anise odor, negative Schäffer's and KOH reactions, and clavate to vesiculose clavate cheilocystidia. It is presented with detailed descriptions, photographs, phylogenetic reconstruction and a comparison with phenetically similar species.

Serological evidence of chronic pulmonary Aspergillosis in tuberculosis patients in Kenya

BackgroundPulmonary tuberculosis (PTB) is a significant risk factor for fungal infection. The cavitary lesions post PTB serves as a good reservoir for fungal colonization and subsequent infection. Furthermore, the severe immunosuppression associated with HIV and TB co-infection is another predisposition. The inadequate capacity to investigate and manage fungal infection in PTB patients increases their morbidity and mortality. The study aimed to provide serological evidence of chronic pulmonary aspergillosis (CPA) among PTB patients in Kenya. Towards this, we analysed 234 serum samples from patients presenting with persistent clinical features of PTB infections despite TB treatment in four referral hospitals.MethodsThis was a cross sectional laboratory based study and patients were recruited following an informed consent. Serological detection of Aspergillus fumigatus IgG was done using enzyme-linked immunosorbent assay (Bordier Affinity Products SA). Sputum samples were subjected to microscopy and standard fungal culture. The isolated fungi were subjected to macro and micro morphological identifications and confirmed by sequence analysis of calmadulin, betatubilin and ITS genes.ResultsSerological evidence of CPA or fungal sensitization was 46(19.7%) and equivocal or borderline was 14(6.0%). Mycological investigations of sputum resulted in 88(38%) positive for fungal culture. Aspergillus spp. accounted for 25(28%) of which A. fumigatus was 13(14.8%), A. niger 8(9.1%), A. terreus, A. flavus, A. candidus and A. clavatus 1 (1.1%) each. This was followed by Penicillium spp. 10 (11.4%), Scedosporium spp. 5 (5.7%) and Rhizopus spp. 3 (3.4%). Among the yeasts; Candida albicans accounted for 18(20.5%) followed by C. glabrata 5(5.7%). Cryptococcus spp. was isolated from 3(3.4%) of the samples while 13(14.8%) were other yeasts.ConclusionChronic pulmonary aspergillosis is a significant co-morbidity in PTB patients in Kenya that could be misdiagnosed as relapse or treatment failures in the absence of reliable diagnostic and clinical management algorithm. It could be the cause of persistent clinical symptoms despite TB treatment often misdiagnosed as TB smear/GeneXpert MTB/RIF® negative or relapse. We recommend that all patients with persistent clinical symptoms despite TB treatment should be subjected to fungal investigations before retreatment.

Talaromyces santanderensis: A New Cadmium-Tolerant Fungus from Cacao Soils in Colombia.

Inorganic pollutants in Colombian cocoa (Theobroma cacao L.) agrosystems cause problems in the production, quality, and exportation of this raw material worldwide. There has been an increased interest in bioprospecting studies of different fungal species focused on the biosorption of heavy metals. Furthermore, fungi constitute a valuable, profitable, ecological, and efficient natural soil resource that could be considered in the integrated management of cadmium mitigation. This study reports a new species of Talaromyces isolated from a cocoa soil sample collected in San Vicente de Chucurí, Colombia. T. santanderensis is featured by Lemon Yellow (R. Pl. IV) mycelium on CYA, mono-to-biverticillade conidiophores, and acerose phialides. T. santanderensis is distinguished from related species by its growth rate on CYAS and powdery textures on MEA, YES and OA, high acid production on CREA and smaller conidia. It is differentiated from T. lentulus by its growth rate on CYA medium at 37 °C without exudate production, its cream (R. PI. XVI) margin on MEA, and dense sporulation on YES and CYA. Phylogenetic analysis was performed using a polyphasic approach, including different phylogenetic analyses of combined and individual ITS, CaM, BenA, and RPB2 gene sequences that indicate that it is new to science and is named Talaromyces santanderensis sp. nov. This new species belongs to the Talaromyces section and is closely related to T. lentulus, T. soli, T. tumuli, and T. pratensis (inside the T. pinophilus species complex) in the inferred phylogeny. Mycelia growth of the fungal strains was subjected to a range of 0–400 mg/kg Cd and incorporated into malt extract agar (MEA) in triplicates. Fungal radial growth was recorded every three days over a 13-day incubation period and In vitro cadmium tolerance tests showed a high tolerance index (0.81) when the mycelium was exposed to 300 mg/kg of Cd. Results suggest that T. santanderensis showed tolerance to Cd concentrations that exceed the permissible limits for contaminated soils, and it is promising for its use in bioremediation strategies to eliminate Cd from highly contaminated agricultural soils.

A phylogenetic perspective of antiviral species of the genus Artemisia (Asteraceae-Anthemideae): A proposal of anti SARS-CoV-2 (COVID-19) candidate taxa

IntroductionDifferent classes of disease-causing viruses are widely distributed universally. Plant-based medicines are anticipated to be effective cures for viral diseases including the COVID-19, instigated by severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2). This study displays the phylogenetic perspective of Artemisia and proposes some candidate taxa against different viral diseases, including SARS-CoV-2. MethodsData of Artemisia with antiviral activity were obtained from different published sources and electronic searches. A phylogenetic analysis of the nrDNA ITS sequences of reported antiviral Artemisia species, along with the reference species retrieved from the NCBI GenBank database, was performed using the maximum likelihood (ML) approach. ResultsIn total, 23 Artemisia species have been documented so far with antiviral activity for 17 different types of viral diseases. 17 out of 23 antiviral Artemisia species were included in the ITS phylogeny, which presented the distribution of these antiviral Artemisia species in clades corresponding to different subgenera of the genus Artemisia. In the resultant ML tree, 10 antiviral Artemisia species appeared within the subgenus Artemisia clade, 2 species appeared within the subgenus Absinthium clade, 3 species appeared within the subgenus Dracunculus clade, and 2 species appeared within the subgenus Seriphidium clade. DiscussionArtemisia species from different subgenera with antiviral activity are prevalent in the genus, with most antiviral species belonging to the subgenus Artemisia. A detailed analysis of taxa from all subgenera, particularly the subgenus Artemisia, is therefore proposed in order to discover compounds with potential anti-SARS-CoV-2 activity.

New Records of the Psychrophilic Tetracladium Species Isolated from Freshwater Environments in Korea

We investigated the diversity of aquatic fungi from freshwater environments in Korea. From the survey of aquatic fungi, three Tetracladium (Leotiomycetes, Helotiales) strains, NNIBRFG814, NNIBRFG1997, and NNIBRFG3891, were isolated from decaying leaves of Fraxinus rhynchophylla located in Taebaek-si, Gangwon-do; freshwater foam in the freshwater of the Gam stream in Gimcheon-si; and from filtered freshwater of the Yangsan stream in Mungyeong-si, Gyeongsangbuk-do, respectively. The isolated fungal strains were identified using ITS sequence analysis and morphological characters. A sequence analysis indicated that the fungal isolates were closely related to Tetracladium breve MK371730 (100%, NNIBRFG814), Tetracladium setigerum KU519120 (98.96%, NNIBRFG1997), and T. marchalianum MK353126 (98.14%, NNIBRFG3891). In terms of growth conditions, it was confirmed that NNIBRFG814 grew best at 20 °C, did not grow above 30 °C, and grew at 5 °C as a psychrophilic fungus. NNIBRFG814 was included in the T. breve clade but its morphology was not similar to that of T. breve. Thus, NNIBRFG814 is a new species, and T. fraxineum sp. nov. NNIBRFG1997 and NNIBRFG3891 are the first records in Korea of T. setigerum and T. marchalianum, respectively.

P245 Arthrinium species, a filamentous ascomycetes isolated from samples of human cutaneous infections-report from a medical mycology laboratory of Assam, North-East India

Poster session 2, September 22, 2022, 12:30 PM - 1:30 PMObjectivesThis study aims to report the isolation of closely related Arthrinium species from superficial skin lesions of five cases from a medical mycology laboratory of Assam, North East India.MethodsThe lesions were decontaminated with 70% ethanol and skin scrapings were collected on a sterilized glass plate. Direct mounts were prepared in 10%-20% KOH and cultures were put in Saboraud's Dextrose Agar with antibiotics, 5% sheep blood agar, and dermatophyte test medium (Himedia, India). Plates and tubes were incubated as per standard mycological techniques described. Molecular identification was done using ITS sequence analysis using ITS1 and ITS4 universal primers.ResultsDirect mount showed presence of hyphae with arthrospores in 3/5 cases. In one case, fungal hyphae was seen along with spore-like oval or round structures of about 3-4 μm diameter. Pure growth was seen after 7-14 days in multiple culture tubes in all five cases. Colonies were white, downy initially becoming white, and floccose on further incubation. Subculture on PDA in all the cases for 15-20 days revealed black, round, and oval spores of 3-5 μm suggesting Arthrinium spp.The taxonomical identification was done by constructing a phylogenetic tree of the ITS sequences of the Arthrinium isolates of this study along with reference Arthrinium strains and Seiridium phylicae as the outgroup taxa.The phylogenetic analysis clustered the isolates of this study into closely related Arthrinium species.ConclusionThe genus Arthrinium belonging to the family Apiosporaceae, class Sordariomycetes which comprises of a group of filamentous ascomycetes fungi is rarely reported from human infections. We are reporting closely related Arthrinium spp from five cases of skin lesions from Assam, North East India. Three of the 5 cases hailed from tea garden areas of Assam. Arthrinium isolation in clinically significant cases and in multiple tubes may not be disregarded as a contaminant.

Distribution, cytotoxicity, and antioxidant activity of fungal endophytes isolated from Tsuga chinensis (Franch.) Pritz. in Ha Giang province, Vietnam

PurposeAn endangered Tsuga chinensis (Franch.) Pritz. is widely used as a natural medicinal herb in many countries, but little has been reported on its culturable endophytic fungi capable of producing secondary metabolites applied in modern medicine and pharmacy. The present study aimed to evaluate the distribution of fungal endophytes and their cytotoxic and antioxidant properties.MethodsThis study used the surface sterilization method to isolate endophytic fungi which were then identified using morphological characteristics and ITS sequence analysis. The antimicrobial and cytotoxic potentials of fungal ethyl acetate extracts were evaluated by the minimum inhibitory concentration (MIC) and sulforhodamine B (SRB) assays, respectively. Paclitaxel-producing fungi were primarily screened using PCR-based molecular markers. Additionally, biochemical assays were used to reveal the antioxidant potencies of selected strains.ResultsA total of sixteen endophytic fungi that belonged to 7 known and 1 unknown genera were isolated from T. chinensis. The greatest number of endophytes was found in leaves (50%), followed by stems (31.3%) and roots (18.7%). Out of 16 fungal strains, 33.3% of fungal extracts showed significant antimicrobial activities against at least 4 pathogens with inhibition zones ranging from 11.0 ± 0.4 to 25.8 ± 0.6 mm. The most prominent cytotoxicity against A549 and MCF7 cell lines (IC50 value < 92.4 μg/mL) was observed in Penicillium sp. SDF4, Penicillium sp. SDF5, Aspergillus sp. SDF8, and Aspergillus sp. SDF17. Out of three key genes (dbat, bapt, ts) involved in paclitaxel biosynthesis, strains SDF4, SDF8, and SDF17 gave one or two positive hits, holding the potential for producing the billion-dollar anticancer drug paclitaxel. Furthermore, four bioactive strains also displayed remarkable and wide-range antioxidant activity against DPPH, hydroxyl radical, and superoxide anion, which was in relation to the high content of flavonoids and polyphenols detected.ConclusionThe present study exploited for the first time fungal endophytes from T. chinensis as a promising source for the discovery of new bioactive compounds or leads for the new drug candidates.

Two new species of Botryosphaeriaceae (Botryosphaeriales) and new host/geographical records

Botryosphaeriaceae is the largest family in Botryosphaeriales, consisting of 22 genera. The family is worldwide in distribution and occurs on a wide variety of plant hosts. Botryosphaeriaceae includes endophytic, saprobic, and plant pathogenic species associated with canker and die-back diseases, most common on woody plants. The present study establishes two novel species of Dothiorella and new host and geographical records for species of Botryosphaeria, Dothiorella, Neodeightonia, and Sphaeropsis. Taxa were identified based on morphology coupled with phylogenetic analyses based on maximum likelihood and Bayesian analyses of SSU, LSU, ITS, tef1-α and β-tub sequences. The taxa are described and illustrated.

Cadophora species from marine glaciers in the Qinghai-Tibet Plateau: an example of unsuspected hidden biodiversity

Large numbers of marine glaciers in the Qinghai-Tibet Plateau are especially sensitive to changes of climate and surface conditions. They have suffered fast accumulation and melting and retreated quickly in recent years. In 2017, we surveyed the cold-adapted fungi in these unique habitats and obtained 1208 fungal strains. Based on preliminary analysis of ITS sequences, 41 isolates belonging to the genus Cadophora were detected. As one of the most frequently encountered genera, the Cadophora isolates were studied in detail. Two phylogenetic trees were constructed: one was based on the partial large subunit nrDNA (LSU) to infer taxonomic placement of our isolates and the other was based on multi-locus sequences of LSU, ITS, TUB and TEF-1α to investigate more exact phylogenetic relationships between Cadophora and allied genera. Combined with morphological characteristics, nine Cadophora species were determined, including seven new to science. Among the new species, only C. inflata produces holoblastic conidia and all the others express phialidic conidiogenesis. All isolates have optimum growth temperature at 20 °C or 25 °C. With more species involved, the currently circumscribed genus became obviously paraphyletic. All members are clustered into two main clades: one clade mainly includes most of the Cadophora species which have phialidic conidiogenesis and we refer to as ‘Cadophora s. str.’; the remaining Cadophora species have multiform conidiogenesis and are clustered in the second clade, with members of other genera in Ploettnerulaceae interspersed among the subclades. The results show a high diversity of Cadophora from marine glaciers in the Qinghai-Tibet Plateau and most of them are novel species.

Sulla Powdery Mildew: Phylogeny and Host Range

Sulla is a biannual forage legume cultivated throughout the Mediterranean Basin. It can be severely damaged by powdery mildew, but there is little understanding on its causal species or its host range. The taxonomic characterization of Erysiphe species is mainly based on the examination of chasmothecia morphology, or on the molecular analysis of ITS1 and ITS2 sequences. However, a description of chasmothecia morphology or ITS sequences is not always available to clearly assign a given isolate to an existing or novel Erysiphe species. In an attempt to clarify the identity of the powdery mildew infecting sulla crop we studied the morphology and ITS of nine populations collected over years and countries and compared them with available ITS sequences. Phylogenetic analysis showed that the powdery mildews collected on sulla clustered together with isolates collected on Coronilla varia in an independent clade between the E. guarinonii/E. trifoliorum and E. palczewskii clades, strongly suggesting that all these sequences correspond to the same Erysiphe species. Measurements of conidia and conidial foot cells of the sulla-infecting powdery mildews partially overlap those of other Erysiphe species, although they clearly differentiate from them, supporting the assumption that powdery mildews from sulla belong to a distinct species as observed from the phylogenetic analysis. As far as we know, our study is the first to report the molecular characterization of powdery mildew isolated on plants from the Hedysarum clade. Cross-inoculations confirmed a high specialization of the powdery mildew of sulla, with the sulla’s isolates infecting only the S. coronaria accessions, and none of the accessions from the other legume genera studied. All studied S. coronaria accessions were heavily infected by the sulla isolate but not by any other isolate used. All this points to E. hedysari as the causal agent of the S. coronaria powdery mildew.

Fruit Microbial Communities of the Bisucciu Sardinian Apricot Cultivar (Prunus armeniaca L.) as a Reservoir of New Brewing Starter Strains

Local fruit cultivars may improve the originality of specialty beers both directly, by conferring peculiar tastes and flavors, and indirectly, as a reservoir of new starter strains. Accordingly, the fungal and bacterial communities of Bisucciu fruit, a Sardinian apricot cultivar used to produce a local fruit beer, were here investigated by culture-dependent methods. From the 16S rDNA and ITS sequence analyses of 68 epiphytic isolates, 5 bacterial species and 19 fungal species were identified. Aureobasidium pullulans and Rhodotorula glutinis were the dominant fungal species, while Enterococcus mundtii (Firmicutes) and Frigoribacterium faeni (Actinobacteria) were the most represented species among bacterial isolates. Enrichment cultures of fresh apricot puree, followed by fermentation trials in beer wort and beer wort added with apricot puree, allowed the selection of four isolates of Pichia kudriavzevii, Hanseniaspora uvarum, H. pseudoguilliermondii, and H. clermontiae able to dominate over the Bisucciu native microbiota and to produce from 0.57% to 0.74% (vol/vol) of ethanol. HS-SPME-GC/MS analysis highlighted a significant increase in the ester and alcohol fractions as well as a reduction in terpenes after fermentation with the selected yeasts. Results obtained suggest that the yeast isolates may contribute to the definition of the taste and flavor of beers when used in mixed fermentations with Saccharomyces.

New combination in the genus Hohenbuehelia (Pleurotaceae)

We studied the type specimens of Pleurotus komarnitzkyi to clarify its taxonomic status. Unlike other species in this genus which grow on wood, P. komarnitzkyi is ecologically distinct as it grows at the base of woody stems of desert herbaceous plants. In addition, its fruiting bodies are small in size, which is not typical for the Pleurotus species. Substrate preferences and morphology of fruiting bodies of P. komarnitzkyi resemble those of Hohenbuehelia species. Based on an analysis of ITS and LSU nrDNA sequences, we propose a new combination in the genus Hohenbuehelia.

Erysiphe iranica sp. nov. on Onobrychis caput-galli in Iran

In April 2014, powdery mildew symptoms were found on Onobrychis caputgalli in Khan Ahmad, Gachsaran, Iran. Morphological characters and analysis of ITS and 28S rDNA sequences revealed that this powdery mildew does not match previously recorded species on fabaceous hosts. It is proposed as a new species, Erysiphe iranica, and is described and illustrated, and compared with previous species of powdery mildew recorded on Fabaceae.

Taxonomy and phylogeny of Paramarasmius gen. nov. and Paramarasmius mesosporus, a worldwide distributed fungus with a strict ecological niche

Specimens of Marasmiellus mesosporus from various parts of the world were morphologically and phylogenetically studied. The results confirmed its identity with M. dunensis, described from southern Europe, and M. arenaceus, described from Japan. Marasmiellus mesosporus is a fungus connected with coastal sandy dunes having a wide distribution being collected in southeastern North America, Lesser and Greater Antilles, Pacific Islands, southern Europe and South and East Asia. Its comparison with phylogenetically and morphologically related species is provided. A new genus Paramarasmius containing P. palmivorus, as the type species, and P. mesosporus is described on the base of phylogenetic analyses of ITS and LSU rDNA sequences. A new combination Collybiopsis carneopallida is also proposed.

Genetic Characterization of Microsporum canis Clinical Isolates in the United States.

Microsporum canis is the primary agent causing dermatophytosis in cats, and also infects humans, dogs, and other species. Assessment of genetic variation among M. canis isolates in the United States has not been conducted. Further, M. canis mating type and assessment of disease severity associated with genotypic characteristics have not been rigorously evaluated. We therefore isolated M. canis from 191 domestic cats across the US and characterized genotypes by evaluation of ITS sequence, MAT locus, and microsatellite loci analysis. The genes SSU1 and SUB3, which are associated with keratin adhesion and digestion, were sequenced from a subset of isolates to evaluate potential genetic associations with virulence. Analysis of microsatellite makers revealed three M. canis genetic clusters. Both clinic location and disease severity were significant predictors of microsatellite variants. 100% of the M. canis isolates were MAT1-1 mating gene type, indicating that MAT1-2 is very rare or extinct in the US and that asexual reproduction is the dominant form of replication. No genetic variation at SSU1 and SUB3 was observed. These findings pave the way for novel testing modalities for M. canis and provide insights about transmission and ecology of this ubiquitous and relatively uncharacterized agent.

Phylogenetic and morphological characteristics of Uromycladium falcatariae, the fungal pathogen that causes gall rust epidemics of Falcataria moluccana in Indonesia

AbstractCurrently, gall rust is the most important disease of Falcataria moluccana in Indonesia. The gall rust pathogen was identified as Uromycladium falcatariae, in a recent study using molecular approaches. However, the molecular study of the F. moluccana gall rust pathogen from Indonesia has not been reported. Therefore, the study aimed to analyse this through molecular phylogenetic of the internal transcribed spacer‐5.8S rDNA (ITS), the cytochrome oxidase 3 (CO3) gene and the gene encoding the large ribosomal subunit (LSU) as well as analyses of its spore dimensions and number of striae. For this purpose, isolates of the gall rust pathogen of F. moluccana were collected from three provinces in Java and one province in Sumatra. The average of spore lengths and the widths varies, with the isolate from Bengkulu, having the largest spore dimension. For the number of striae, isolate from Central Java had a wider range compared with other isolates. Phylogenetic analysis of ITS, LSU and CO3 sequences using maximum likelihood revealed a genetically distinct group of the gall rust pathogen of F. moluccana originating from the various provinces in Indonesia with isolates from the Philippines and Timor Leste. The gall rust pathogens may have evolved to adapt to a populations of F. moluccana in Java.

Lepiota sindhudeltana sp. nov. (Agaricaceae; Basidiomycota) from Punjab, Pakistan

Lepiota sindhudeltana sp. nov. was found on sandy soil under Saccharum bengalense during fungal surveys to the deltas of the Indus River and the banks of the River Ravi, Punjab, Pakistan. The new species belongs to section Ovisporae, subsection Felininae and is characterized by dull orange appressed squamules on the pileus, thin pileal context, pale yellow lamellae, a stipe rounded to slightly oblong at base, basidiospores broadly ellipsoid to oblong, neither dextrinoid nor amyloid, short, clavate erect pileal hyphae, and narrowly clavate terminal hyphae of the stipitipellis. The closest described species to the new taxon in the phylogenetic tree, based on analysis of ITS and LSU sequences, is L. angusticystidiata. We present the new species with illustrated morphological description, phylogeny, and comparison with related or similar species.

Occurrence and molecular characterization of Enterocytozoon bieneusi in water buffaloes (Bubalus bubalis) in Turkey

Microsporidia are obligate intracellular fungus-like parasites that infect humans and animals worldwide. However, there is limited epidemiological data on the occurrence and molecular diversity of microsporidia in buffaloes worldwide. In the present study, fecal samples of 300 water buffaloes (Bubalus bubalis) in Kayseri, Sivas, and Samsun provinces of Turkey were investigated using two nested PCR assays targeting the rRNA of E. bieneusi and Encephalitozoon spp. All the fecal samples from water buffalo were found to be negative for Encephalitozoon spp. PCR positive isolates of E. bieneusi were bidirectionally sequenced for genotyping and phylogenetic analyses. Enterocytozoon bieneusi was the only microsporidian species identified in 8 water buffaloes with an overall molecular prevalence of 2.7%. Two known genotypes, YNDCEB-90 (n = 5) and J (n = 3) were identified by ITS sequence analysis. The YNDCEB-90 and J genotypes fall into zoonotic Group 1 and 2 of E. bieneusi in the phylogenetic tree, respectively. These findings suggested that water buffalo in Turkey are harbouring zoonotic genotypes of E. bieneusi and may have a significant risk for zoonotic transmission to humans. This is the first report of detecting E. bieneusi genotypes J and YNDCEB-90 in water buffaloes. Further insight into the epidemiology of E. bieneusi in water buffaloes in different geographical areas in Turkey will be highly important to have determined the public health significance of this pathogen.