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Background The nasopharyngeal bacterial and fungal microbiota of normal horses and those with nasopharyngeal cicatrix syndrome (NCS) are unknown. Hypotheses/objectives To describe the microbiota from nasopharyngeal washes of healthy horses and of horses acutely affected with NCS. Animals Twenty-six horses acutely affected with NCS horses and 14 unaffected horses. Methods Prospective, observational cohort study. Horses were recruited by investigators through personal communications in central Texas. Bacterial (16s RNA) and fungal (internal transcribed spacer) microbiota from nasopharyngeal washes were evaluated. Polymerase chain reaction for detection of Pythium insidiosum was performed. Results Results indicated that 6 fungal genera (Alternaria, Bipolaris, Microascus, Spegazzinia, Paraconiothyrium, Claviceps) and 1 bacterial genera (Staphylococcus) were significantly different between affected and unaffected horses. The fungal genus Bipolaris had increased abundance in NCS affected horses and on NCS affected farms. Pythium insidiosum was absent in the nasopharyngeal wash of all horses, irrespective of health status. Conclusion and clinical importance Significant differences were identified in the fungal microbiota in horses affected with NCS and farms affected with NCS compared to those unaffected. Therefore, Bipolaris warrants further investigation.

Species of the genus Microascus are uncommon agents of human diseases despite their ubiquitous presence in the environment. In this communication, the first case of white grain eumycetoma caused by the fungus Microascus gracilis is reported. The patient was initially misdiagnosed as having actinomycetoma based on the grains morphological and cytological features and was treated with antimicrobial therapy with no clinical improvement. She underwent wide local surgical excision to improve the response to medical treatment and further grain cultural, molecular and taxonomy techniques were conducted and the diagnosis of mycetoma due to M. gracilis was established. The antifungal susceptibilities of this isolate to nine drugs were tested in vitro and they showed poor activity. Combination therapy with surgery and itraconazole led to complete recovery. A medical literature search revealed no previous report on M. gracilis as a causative agent of eumycetoma and hence we are reporting this new causative agent of human eumycetoma. Also, the difficulty in the management of this patient emphasizes the need for accurate and appropriate diagnostic tests for the identification of mycetoma-causative organisms and thus proper management.

Karst caves are characterized by darkness, low temperature, high humidity, and oligotrophic organisms due to its relatively closed and strongly zonal environments. Up to now, 1626 species in 644 genera of fungi have been reported from caves and mines worldwide. In this study, we investigated the culturable mycobiota in karst caves in southwest China. In total, 251 samples from thirteen caves were collected and 2344 fungal strains were isolated using dilution plate method. Preliminary ITS analyses showed that these strains belonged to 610 species in 253 genera. Among these species, 88.0% belonged to Ascomycota, 8.0% Basidiomycota, 1.9% Mortierellomycota, 1.9% Mucoromycota, and 0.2% Glomeromycota. The majority of these species have been previously known from other environments, and some of them are known as mycorrhizal or pathogenic fungi. About 52.8% of these species were discovered for the first time in karst caves. Based on morphological and phylogenetic distinctions, 33 new species were identified and described in this paper. Meanwhile, one new genus of Cordycipitaceae, Gamszarea, and five new combinations are established. This work further demonstrated that Karst caves encompass a high fungal diversity, including a number of previously unknown species. Taxonomic novelties: New genus: Gamszarea Z.F. Zhang & L. Cai; Novel species: Amphichorda cavernicola, Aspergillus limoniformis, Aspergillus phialiformis, Aspergillus phialosimplex, Auxarthron chinense, Auxarthron guangxiense, Auxarthronopsis globiasca, Auxarthronopsis pedicellaris, Auxarthronopsis pulverea, Auxarthronopsis stercicola, Chrysosporium pallidum, Gamszarea humicola, Gamszarea lunata, Gamszarea microspora, Gymnoascus flavus, Jattaea reniformis, Lecanicillium magnisporum, Microascus collaris, Microascus levis, Microascus sparsimycelialis, Microascus superficialis, Microascus trigonus, Nigrospora globosa, Paracremonium apiculatum, Paracremonium ellipsoideum, Paraphaeosphaeria hydei, Pseudoscopulariopsis asperispora, Setophaeosphaeria microspora, Simplicillium album, Simplicillium humicola, Wardomycopsis dolichi, Wardomycopsis ellipsoconidiophora, Wardomycopsis fusca; New combinations: Gamszarea indonesiaca (Kurihara & Sukarno) Z.F. Zhang & L. Cai, Gamszarea kalimantanensis (Kurihara & Sukarno) Z.F. Zhang & L. Cai, Gamszarea restricta (Hubka, Kubatova, Nonaka, Cmokova & Řehulka) Z.F. Zhang & L. Cai, Gamszarea testudinea (Hubka, Kubatova, Nonaka, Cmokova & Řehulka) Z.F. Zhang & L. Cai, Gamszarea wallacei (H.C. Evans) Z.F. Zhang & L. Cai.

Two scopulariopsis-like fungi were isolated when surveying fungi in pig farm soils in China. Sexual structures of these fungi were not observed and their conidia had spinous walls. Phylogenetic analysis based on nucleotide sequences of the internal transcribed spacer 1 and 2 and intervening 5.8S ribosomal RNA gene (ITS), large subunit ribosomal RNA gene (LSU), beta-tubulin (tub2) and translation elongation factor 1-alpha (tef1) gene showed that they were new members of the genus Microascus, and the name M. aculeatus and M. spinosporus were introduced. Genetically the two new species clustered in a well-supported clade close to M. longicollis, but differed in producing relatively long branches. Morphologically M. aculeatus could be distinguished by its conidia with sparse but long spines; M. spinosporus resembled three species of the genus Scopulariopsis, S. asperula, S. brevicaulis and S. flava, but was different in the color of colony and conidia, and the form of conidiophores.

The harsh environments of desert areas lead to natural selection of resistant creatures with obvious characteristics. This experiment looked for salt-tolerant fungi from native halophyte plants. Forty fungi isolated from three halophyte plant families that were collected from desert areas of Yazd Province in Iran, and the most tolerant isolates were selected at concentrations of 1, 2, 3, 3.5 and 4 molar sodium chloride. Five selected superior isolates were assigned to the phylum Ascomycota based on internal transcribed spacers sequences and β-tubulin gene, as well as morphological characteristics of the genus and species. Aspergillus terreus showed superiority in terms of enzymes and antibacterial properties than other isolates. Other isolates were Acremonium, Paecilomyces, Microascus and Monosorascus. Aspergillus terreus also showed antifungal effects against Aspergillus fumigatus, a human pathogen.

Purpose. To assess in vitro activities of nine antifungal agents (amphotericin B, fluconazole, voriconazole, itraconazole, posaconazole, caspofungin, micafungin, terbinafine and 5-flucytosine) against 93 strains of rare pathogenic fungi and the combined effects of drug combinations against several multidrug-resistant fungi. Methodology. The broth microdilution method M38-A3 and M27-A4 from the Clinical and Laboratory Standards Institute and the checkerboard method were performed in this study. Results. Low MICs for fluconazole were observed in moulds including Tritirachium oryzae, Exophiala attenuata and yeasts. MICs for amphotericin B>2 µg ml−1 were found among Aspergillus nidulans, Fusarium napiforme, Trichoderma longibrachiatum, Tritirachium oryzae, Cunninghamella bertholletiae, Cunninghamella phaeospora, Conidiobolus coronatus, Exophiala attenuata, Ochroconis mirabilis and Rhinocladiella basitona. Multidrug resistance was observed in Microascus spp., Lomentospora prolificans and Pythium insidiosum. Conclusion. Our study illustrated in vitro drug susceptibilities of some rare pathogenic fungi, which provide data to guide clinical treatment of fungal infections.

Occupational exposure to harmful bioaerosols in industrial environments is a real threat to the workers. In particular, dairy-farm workers are exposed to high levels of fungal bioaerosols on a daily basis. Associating bioaerosol exposure and health problems is challenging and adequate exposure monitoring is a top priority for aerosol scientists. Using only culture-based tools does not express the overall microbial diversity and underestimate the large spectrum of microbes in bioaerosols and therefore the extended fungal profile that farmers are exposed to. The aim of this study was to provide an in-depth characterization of fungal exposure at Eastern Canadian dairy farms using qPCR and high-throughput sequencing methods. Specific primers were used for the quantification of Penicillium/Aspergillus and Aspergillus fumigatus in dairy farms air samples. Illumina Miseq sequencing of the ITS1 region provided sequences for the diversity analyses. The minimum and maximum concentration of Penicillium/Aspergillus ranged from 4.6 × 106 to 9.4 × 106 gene copies/m3 and from 1 × 104 gene copies/m3 to 4.8 × 105 gene copies/m3 for Aspergillus fumigatus, respectively. Differences in the diversity profiles of the five dairy farms support the idea that the novel approach identifies a large number of fungal taxa. The most striking differences include Microascus, Piptoporus, Parastagonospora, Dissoconium, Microdochium, Tubilicrinis, Ganoderma, Ustilago, Phlebia and Wickerhamomyces. The presence of a diverse portrait of fungi in air may represent a health risk for workers who are exposed on a daily basis. The broad spectrum of fungi detected in this study includes many known pathogens like Aspergillus, Acremonium, Alternaria and Fusarium. Adequate monitoring of bioaerosol exposure is necessary to evaluate and minimize risks.

Microascus gracilis is a specie of the genus Microascus in the family of Microascaceae and has been isolated from lung. It has never been reported as the cause of disseminated infection in humans. Herein, we report a fatal case of disseminated Microascus gracilis infection in a 65-year-old man with a history of primary idiopathic pulmonary fibrosis, status-post bilateral lung transplant. His course was complicated by donor lung cultures positive for multiple organisms and persistent pleural effusions. Multiple lung biopsy and bronchial lavage specimens were negative for mold. Later, pleural fluid cultures grew M. gracilis confirmed by DNA sequencing. Despite aggressive antifungal treatment, the patient continued to deteriorate with altered mental status. Imaging showed scattered hemorrhagic and hypodense lesions in the brain. The patient eventually succumbed to his infections and a restricted autopsy was performed. Autopsy findings included multiple hemorrhagic foci and abscesses involving the whole brain. Numerous punctuate, tan-white circular lesions were on the endocardium and diffuse tan exudates covered the pericardium and lungs. Histologically, similar fungal organisms with septate branching hyphae and short chains of conidia were identified, along with hemorrhage, neutrophilic inflammation, and necrosis in the brain, pleura, peripheral parenchyma of lungs and heart. This is the first reported case of disseminated M. gracilis infection in an immunosuppressed human, indicating it can cause localized infections and disseminated infections. This case increases our awareness of such fatal opportunistic infections, particularly in lung transplant patients, and urges earlier aggressive prophylaxis, diagnosis, and treatment.

The gastrointestinal (GI) tract harbors a diverse population of microorganisms. While much work has been focused on the characterization of the bacterial community, very little is known about the fungal community, or mycobiota, in different animal species and chickens in particular. Here, we characterized the biogeography of the mycobiota along the GI tract of day 28 broiler chicks and further examined its possible shift in response to bacitracin methylene disalicylate (BMD), a commonly used in-feed antibiotic, through Illumina sequencing of the internal transcribed spacer 2 (ITS2) region of fungal rRNA genes. Out of 124 samples sequenced, we identified a total of 468 unique fungal features that belong to four phyla and 125 genera in the GI tract. Ascomycota and Basidiomycota represented 90% to 99% of the intestinal mycobiota, with three genera, i.e., Microascus, Trichosporon, and Aspergillus, accounting for over 80% of the total fungal population in most GI segments. Furthermore, these fungal genera were dominated by Scopulariopsis brevicaulis (Scopulariopsis is the anamorph form of Microascus), Trichosporon asahii, and two Aspergillus species. We also revealed that the mycobiota are more diverse in the upper than lower GI tract. The cecal mycobiota transitioned from being S. brevicaulis dominant on day 14 to T. asahii dominant on day 28. Furthermore, 2-week feeding of 55 mg/kg BMD tended to reduce the cecal mycobiota α-diversity. Taken together, we provided a comprehensive biogeographic view and succession pattern of the chicken intestinal mycobiota and its influence by BMD. A better understanding of intestinal mycobiota may lead to the development of novel strategies to improve animal health and productivity. IMPORTANCE The intestinal microbiota is critical to host physiology, metabolism, and health. However, the fungal community has been often overlooked. Recent studies in humans have highlighted the importance of the mycobiota in obesity and disease, making it imperative that we increase our understanding of the fungal community. The significance of this study is that we revealed the spatial and temporal changes of the mycobiota in the GI tract of the chicken, a nonmammalian species. To our surprise, the chicken intestinal mycobiota is dominated by a limited number of fungal species, in contrast to the presence of hundreds of bacterial taxa in the bacteriome. Additionally, the chicken intestinal fungal community is more diverse in the upper than the lower GI tract, while the bacterial community shows an opposite pattern. Collectively, this study lays an important foundation for future work on the chicken intestinal mycobiome and its possible manipulation to enhance animal performance and disease resistance.

A 60-year-old woman presented with a nodular granulomatous skin lesion on her right thumb. It had developed after inoculation of a splinter of wood. Because it was resistant to various therapies, the nodule was finally excised. Complete healing followed this surgery and a melanised filamentous fungus with scopulariopsis-like morphology was recovered from the dermal tissue. Fitting with no known species, the fungus was subjected to extensive morphological, physiological and genetic investigations. It was characterised by resistance to cycloheximide, growth at 37°C, branched conidiophores with cylindrical annellides in brush-like groups producing dark conidia in basipetal chains, and cleistothecia with ellipsoidal to slightly reniform ascospores. Genetically it clustered in a well-supported clade together with Microascus (M.) brunneosporus, Microascus chinensis, Microascus intricatus, Microascus longicollis, Microascus micronesiensis and Microascus onychoides, but formed an independent branch distant from the other Microascus species. Based on its unique genetic characteristics and morphological findings, the isolate is proposed as a new species, Microascus ennothomasiorum. Morphologically it differs from its phylogenetically closest species by its branched conidiophores and ascomata with a peridium of textura intricata. Our observation once again emphasises that dermal granulomas can be caused by uncommon fungi; diagnostics should therefore include appropriate mycological investigations.

Karst caves are distinctly characterised by darkness, low to moderate temperatures, high humidity, and scarcity of organic matter. During the years of 2014–2015, we explored the mycobiota in two unnamed Karst caves in Guizhou province, China, and obtained 563 fungal strains via the dilution plate method. Preliminary ITS analyses of these strains suggested that they belonged to 246 species in 116 genera, while 23.5 % were not identified to species level. Among these species, 85.8 % (211 species) belonged to Ascomycota; 7.3 % (18 species) belonged to Basidiomycota; 6.9 % (17 species) belonged to Mucoromycotina. The majority of these species have been previously known from other environments, mostly from plants or animals as pathogens, endophytes or via a mycorrhizal association. We also found that 59 % of these species were discovered for the first time from Karst caves, including 20 new species that are described in this paper. The phylogenetic tree based on LSU sequences revealed 20 new species were distributed in six different orders. In addition, ITS or multi-locus sequences were employed to infer the phylogenetic relationships of new taxa with closely related allies. We conclude that Karst caves encompass a high fungal diversity, including a number of previously unknown species. Novel species described include: Amphichorda guana, Auxarthronopsis guizhouensis, Biscogniauxia petrensis, Cladorrhinum globisporum, Collariella quadrum, Gymnoascus exasperatus, Humicola limonisporum, Metapochonia variabilis, Microascus anfractus, Microascus globulosus, Microdochium chrysanthemoides, Paracremonium variiforme, Pectinotrichum chinense, Phaeosphaeria fusispora, Ramophialophora globispora, Ramophialophora petraea, Scopulariopsis crassa, Simplicillium calcicola, Volutella aeria, and Wardomycopsis longicatenata.

One hundred samples of carpet dusts (50 samples from residential houses and 50 samples from mosques) were surveyed for the presence of the potentially pathogenic fungi Microascus/Scopulariopsis species complex (Microascales:Ascomycota). The samples were collected from different sites in Duhok province,Kurdistan region,Iraq, during September, 2014 to May, 2015. Four species of Microascus (M.brunneosporus, M.cirrosus, M.pyramidus and M.paisii) and three species of Scopulariopsis (S.asperula, S.brevccaulis and S.flava) were identified. Acaulium acremonium (=Scopulariopsis acremonium) was also detected. The diagnostic features of the reported species based on micro-morphological features of their reproductive structures were provided. M.brunneosporus represents a new record for the Iraqi mycobiota. The identified species were reported in several publications as potentially pathogenic to human.

Scopulariopsis-like species are often reported from the indoor environment, as well as from clinical samples. The lack of type isolates and thorough phylogenetic studies in theMicroascaceaehampered the correct identification of these isolates. Based on recent phylogenetic studies, which resulted in multiple name changes, the aim is to molecularly identify theScopulariopsisand scopulariopsis-like species which occur in the indoor environment and give an overview of the current species in these genera and their habitats. Strains from the CBS culture collection were supplemented with almost 80 indoor strains of which the internal transcribed spacer 1 and 2 and intervening 5.8S nrDNA (ITS), beta-tubulin (tub2) and translation elongation factor 1-alpha (tef1) gene regions were sequenced for phylogenetic inference. The multi-gene phylogenies recognise 33Microascusspecies and 12Scopulariopsisspecies and showed that the recently established genusFuscoannellis, typified byScopulariopsis carbonaria, should be synonymized with the genusYunnania. Seven newMicroascusspecies, four newScopulariopsisspecies, and one newYunnaniaspecies, are described, and a new name inMicroascusand two new name combinations (one inMicroascus, and one inYunnania) are proposed. In the indoor environment 14Microascusspecies and threeScopulariopsisspecies were found.Scopulariopsis brevicaulis(22 indoor isolates) andMicroascus melanosporus(19 indoor isolates) are the most common indoor species, in number of isolates, followed byM. paisii(8 indoor isolates) andS. candida(7 indoor isolates). A genus phylogeny based on the ITS,tef1and the large subunit 28S nrDNA (LSU) of the type or representative isolates of all here recognised species is provided depicting all species habitats. No correlation between phylogenetic relationship and habitat preference could be observed. Ten species which are found indoor are also found in relation with human-derived samples. A table showing recent name changes and a key to common species ofScopulariopsisand scopulariopsis-like genera found indoors is included.

One hundred samples of carpet dusts (50 samples from residential houses and 50 samples from mosques) were collected from different sites in Duhok province during the year 2014 to 2015 for the objective of the study of the occurrence of keratinolytic and other potentially pathogenic fungi using hair baiting method. A total of 24 fungal species (17 species from house dust) and (12 species from mosque dust) in addition to non-sporulating mycelia and yeasts were isolated and identified. The keratinolytic species Arthroderma cuniculi, Chrysosporium tropicum and Gymnoascus ressii were detected. Potentially pathogenic fungi in the genera Aspergillus, Arthrographis,Geomyces, Microascus,Scopulariopsis and Neoscytalidium were also able to colonize and grow on baited hairs.

The current knowledge of invasive Scopulariopsis/Microascus infection in lung transplantation has been derived from only four case reports. Although these fungi are uncommon compared with Aspergillus, they are highly resistant to the current antifungal agents, and the mortality is extremely high. To explore the risk factors, clinical manifestations, notable diagnostic characteristics and outcomes of positive Scopulariopsis/Microascus isolation in lung transplantation patients. We included all cases with positive Scopulariopsis/Microascus isolation from lower respiratory tracts or bronchial mucosa biopsies in our lung transplantation centre. Proven cases from the literature were added. Positive isolation occurred in 2% (3/157) in our centre. Four cases from the literature were added. The mortality could be considered as high as 80%, once the two cases of colonisation were excluded. The average interval between transplantation and positive isolation was 106 (19-131) days. A total of 57.1% of patients had experienced a combination of infection with Aspergillus or other fungi as well as long-term azole antifungal agent treatment before the positive isolation, which may be possible risk factors. The combination of micafungin, posaconazole and terbinafine may be an effective treatment. The peak time of positive isolation was consistent with that of some opportunistic pathogens, and the possible risk factors were the infection of other fungi as well as prior long-term azole antifungal administration. In addition to its high mortality, Scopulariopsis/Microascus was also highly resistant to common antifungal agents and the combination of two or three drugs for therapy was recommended.

Dematiaceous (melanized or phaeoid) fungi are common in the environment. However, clinical diseases caused by them are uncommon. Despite their rarity, they are being increasingly recognized as causal agents of disease in man and animal. There is growing awareness among medical fraternity about the clinical significance of these melanized fungi in medical practice. In this article, we have reviewed some clinically significant publications reporting some emerging phaeoid genera up to 2017. The genera reviewed are Alternaria, Aureobasidium, Bipolaris, Exserohilum, Curvularia, Ochroconis, Exophiala, Phialophora, Chaetomium, Neoscytalidium, Leptosphaeria, Microascus, Lecythophora, Phaeoacremonium, Scedosporium, Veronaea, Fonsecaea, Wallemia sebi, Verruconis, etc. In addition, laboratory diagnosis and future areas of research have also been dealt along with conclusion.

Scopulariopsis is a common fungus in the environment, characterized by its intrinsic resistance to the available antifungal drugs. Around 70 cases of infection by this fungus have been described in the literature. Pulmonary and disseminated infections are the most common and their treatment is difficult; therefore, very diverse approaches have been taken, with varied results. A successful outcome has been reported in only a few cases, generally attributed to a multitreatment strategy combining medical and surgical procedures that ultimately led to the resection of the infected tissue if possible, identification of the mould, and an aggressive long-term antifungal therapy. Although most of the infections are caused by Scopulariopsis brevicaulis, a few other species have also been linked to these cases, although molecular evidence has not been proven for all of them. On this basis, more knowledge on the epidemiology, presentation, diagnosis, treatment, and prognosis of these unusual infections would improve their management. This review aims to compile the current data on Scopulariopsis infections.

Microascus species are widely distributed and rarely cause invasive infection in humans. Here we report a case of lung Microascus cirrosus infection in an immunocompetent patient with bronchiectasis. While on systemic voriconazole and aerosolized amphotericin B for three months, the patient's overall condition improved. This case report highlights the possibility of rare pathogen infection occurred in a bronchiectasis patient, as well as the importance of accurate diagnosis and individualized therapy of pulmonary Microascus infection.

The genera Microascus and Scopulariopsis comprise species commonly isolated from soil, decaying plant material and indoor environments. A few species are also recognised as opportunistic pathogens of insects and animals, including humans. In the past, the taxonomy of these fungi has been based on morphology only. With the aim to clarify the taxonomy and phylogeny of these fungi, we studied a large set of clinical and environmental isolates, including the available ex-type strains of numerous species, by means of morphological, physiological and molecular analyses. Species delineation was assessed under the Genealogical Phylogenetic Species Recognition (GCPSR) criterion using DNA sequence data of four loci (ITS region, and fragments of rDNA LSU, translation elongation factor 1-α and β-tubulin). The genera Microascus and Scopulariopsis were found to be separated in two distinct lineages. The genus Pithoascus is reinstated and the new genus Pseudoscopulariopsis is erected, typified by P. schumacheri. Seven new species of Microascus and one of Scopulariopsis are described, namely M. alveolaris, M. brunneosporus, M. campaniformis, M. expansus, M. intricatus, M. restrictus, M. verrucosus and Scopulariopsis cordiae. Microascus trigonosporus var. macrosporus is accepted as a species distinct from M. trigonosporus. Nine new combinations are introduced. Microascus cinereus, M. longirostris, P. schumacheri and S. flava are neotypified. A table summarising the morphological features of the species treated and identification keys for each genus are provided.

Purpose Fungi of Scopulariopsis and Microascus genera cause a wide range of infections, with S. brevicaulis being the most prevalent aetiological agent of mould onychomycosis. Proper identification of these pathogens requires sporulating culture, which considerably delays the diagnosis. So far, sequencing of rDNA regions of clinical isolates has produced ambiguous results due to the lack of reference sequences in publicly available databases. Thus, there is a clear need for the development of new molecular methods that would provide simple, rapid and highly specific identification of Scopulariopsis and Microascus species. The objective of this study was to develop simple and fast assays based on PCR and real-time PCR for specific detection of fungi from Scopulariopsis and Microascus genera, and separately, S. brevicaulis species. Methods On the basis of alignment of β-tubulin gene sequences, Microascus/Scopulariopsis-specific primers were designed and S. brevicaulis-specific primers were reevaluated. DNA from cultured fungal isolates, extracted in a two-step procedure, was used in Microascus/Scopulariopsis-specific and S. brevicaulis-specific PCR and real-time PCR followed by electrophoresis or melting temperature analysis, respectively. Results The specificity of the assays was confirmed, as positive results were obtained only for Scopulariopsis spp. and Microascus spp. isolates tested in Microascus/Scopulariopsis-specific assay, and only for S. brevicaulis and S. koningii (syn. S. brevicaulis) isolates in a S. brevicaulis-specific assay, respectively, and no positive results were obtained neither for other moulds, dermatophytes, yeast-like fungi, nor for human DNA. Conclusions The developed assays enable fast and unambiguous identification of Microascus spp. and Scopulariopsis spp. pathogens.

Strains originally affiliated to the genera Scopulariopsis and Microascus were compared regarding the scopularide production in order to investigate their ability to produce the cyclodepsipeptides and select the best suited candidate for subsequent optimisation processes. Phylogenetic calculations using available sequences of the genera Scopulariopsis and Microascus revealed that most of the sequences clustered within two closely related groups, comprising mainly Scopulariopsis/Microascus brevicaulis and Microascus sp., respectively. Interestingly, high yields of scopularide A were exhibited by three strains belonging to S./M. brevicaulis, while lower titres were observed for two strains of Microascus sp. Close phylogenetic distances within and between the two groups supported the proposed combination of both genera into one holomorph group. Short phylogenetic distances did not allow a clear affiliation at the species level on the basis of ribosomal DNA sequences, especially for Microascus sp. strains. Additionally, several sequences originating from strains assigned to Scopulariopsis exhibited a polyphyletic nature. The production pattern is in accordance with the phylogenetic position of the strains and significant production of scopularide B could only be observed for the S./M. brevicaulis strain LF580. Thus, the phylogenetic position marks the biotechnologically interesting strains and matters in optimisation strategies. In conclusion, the ability of all five strains to produce at least one of the scopularides suggests a distribution of the responsible gene cluster within the holomorph group. Setting the focus on the production of the cyclodepsipeptides, strain LF580 represents the best candidate for further strain and process optimisation.

We present a case of brain abscess in an immunocompetent child due to the dematiaceous fungus Microascus cinereus, an organism commonly found in soil and stored grains. The etiologic agent was demonstrated by direct microscopy and culture. The patient responded well to surgical excision of abscess along with a course of amphotericin B and voriconazole. Microascus species have emerged as significant invasive pathogens especially in the immunocompromised patients. To the best of our knowledge, this is the first reported case of brain abscess caused by M. cinereus in an immunocompetent individual with no underlying risk factors.

Until recently, cinematographic film was largely cellulose-triacetate-based. However, this material is highly susceptible to biodeterioration, thus placing historic film collections, an important part of the cultural heritage of many countries, at risk. In the present study, samples taken from several biodeteriorated color cinematographic films belonging to the collection of the Cuban Institute for Cinematographic Industry and Arts (ICAIC) were investigated. Infrared spectroscopy showed that all films were of the same composition, i.e., a gelatin emulsion coating one side of a cellulose-triacetate-based film support. The films were analyzed by environmental scanning electron microscopy and scanning electron microscopy to determine the degree of biodeterioration and the type of colonizing microorganisms. Significant fungal colonization was found on both sides of the films in all samples, with a higher concentration of fungi on the gelatin emulsion side. Epifluorescence microscopy of fluorochrome-dyed films demonstrated that some of the fungi were still active, indicating that the films under study, and probably others at the ICAIC, are at risk of further deterioration. Fungi were identified by molecular biology techniques. The fungi mainly responsible for the observed biodeterioration were those belonging to the genera Aspergillus and Cladosporium, although other genera, such as Microascus and Penicillium, were identified as well. In accordance with the findings described herein, the existing guidelines for the prevention and control of film biodeterioration are discussed.

Submitted by CAMILA SOUZA SANTOS null (cssantos12@gmail.com) on 2016-03-26T15:11:36Z No. of bitstreams: 1 dissertação final camila.pdf: 6327362 bytes, checksum: be4f63024f7b4a50aa4f8f1e10987053 (MD5) Approved for entry into archive by Ana Paula Grisoto (grisotoana@reitoria.unesp.br) on 2016-03-28T14:44:29Z (GMT) No. of bitstreams: 1 santos_cs_me_araiq_par.pdf: 796529 bytes, checksum: 5e16538a32b063bcef781b76b10382c2 (MD5) Made available in DSpace on 2016-03-28T14:44:29Z (GMT). No. of bitstreams: 1 santos_cs_me_araiq_par.pdf: 796529 bytes, checksum: 5e16538a32b063bcef781b76b10382c2 (MD5) Previous issue date: 2016-03-14 O presente trabalho apresenta os resultados obtidos sobre o estudo químico e biológico dos fungos endolíticos Nodulisporium sp. e Microascus sp. associados à alga vermelha Asparagopsis taxiformis. Para tanto, fez-se uso das técnicas cromatográficas de separação CLAE (cromatografia líquida de alta eficiência), CG-EM (cromatografia gasosa acoplada a espectrometria de massas) e cromatografia em coluna (CC), além da avaliação dos potenciais antifúngico, anticolinesterásico e citotóxico dos extratos brutos, frações e subfrações de Nodulisporium sp. e Microascus sp. O estudo químico levou ao isolamento de quatro substâncias pertencentes à classe das diidroisocumarinas (substâncias S1, S2, S10 e S11), duas dicetopiperazinas (substâncias S3 e S4), um composto fenólico (S12), uma isoflavona (S8), um derivado hidroxiimino (S9), uma mistura contendo uma amida (S5) e um diol vicinal (S6), além do isolamento de um composto aromático (substância S7), ainda em fase de identificação. As estruturas das substâncias isoladas foram identificadas com base nos dados espectroscópicos de ressonância magnética nuclear uni e bidimensional (RMN de 1H, RMN de 13C, TOCSY-1D, HSQC, HMBC, COSY e HSQC-TOCSY), UV-Vis e espectrometria de massas (EM). Comporta ressaltar que o diol vicinal (S6) está sendo descrito pela primeira vez na literatura e que as demais substâncias isoladas não haviam sido relatadas para os gêneros Nodulisporium ou Microascus. A avaliação das atividades antifúngica e anticolinesterásica evidenciaram o potencial de bioatividade de extratos e frações de Nodulisporium sp. e Microascus sp, sendo reveladas atividades forte e moderada frente às enzimas acetilcolinesterase humana e de enguia elétrica. Este é o primeiro estudo químico e biológico relatado na literatura para os endófitos Nodulisporium sp. e Microascus sp. associados à alga vermelha Asparagopsis taxiformis e seus resultados promissores destacam a elevada quimiodiversidade dos extratos e frações dos fungos endofíticos obtidos do ambiente marinho, bem como seu potencial de bioatividade. Evidenciam ainda a importância de se aprofundar os estudos destas linhagens fúngicas visando contribuições relevantes na busca de protótipos para novos agentes terapêuticos através da exploração sustentável da biodiversidade marinha brasileira. This work presents results from the chemical and biological study of endophytic fungal strains of Nodulisporium sp and Microascus sp associated to the red alga Asparagopsis taxiformis. Chromatographic separation techniques as HPLC, GC-MS and CC have been used in addition to bioassays for evaluation of the antifungal, anticholinesterasic and cytotoxic potential of crude extracts, fractions and subfractions of Nodulisporium sp and Microascus sp fungal strains. The chemical study led to the isolation of four dihydroisocoumarins (compounds S1, S2, S10 and S11), two diketopiperazines (compounds S3 and S4), one phenolic compound (S12), one isoflavone (S8), one hydroxyl-imino derivative (S9), a mixture containing one amide (S5) and one novel vicinal diol (S6), in addition to one aromatic compound (compounds S7) still under structural investigation. Their structures were identified based on 1D and 2D NMR spectroscopic data (1H and 13C NMR, TOCSY-1D, HSQC, HMBC, COSY and HSQC-TOCSY), UV-Vis and mass spectrometry (MS). Compound S6 is an aliphatic vicinal diol and is herein described as a novel compound whereas the remaining compounds had not been described before neither for Nodulisporium nor Microascus genera. Antifungal and anticholinesterase activities evaluation of extracts and fractions of Nodulisporium sp e Microascus sp evidenced their strong and moderate potential against human and electric eel acetylcholinesterase enzymes. This is the first report on the chemical and biological studies of endophytic fungi Nodulisporium sp and Microascus sp, associated to the red algae Asparagopsis taxiformis. Their promising results highlight the outstanding chemodiversity of extracts and fractions from such fungal strains from marine environment as well as their bioactivity potential. They also evidence the importance of further investigating such fungal strains aiming at relevant contributions in the search for novel therapeutic agents through sustainable exploration of Brazilian marine biodiversity.

The taxonomy of the synnematous genera Cephalotrichum, Doratomyces and Trichurus, and other related genera Gamsia, Wardomyces and Wardomycopsis, has been controversial and relies mainly on morphological criteria. These are microascaceous saprobic fungi mostly found in air and soil and with a worldwide distribution. In order to clarify their taxonomy and to delineate generic boundaries within the Microascaceae, we studied 57 isolates that include clinical, environmental and all the available ex-type strains of a large set of species by means of morphological, physiological and molecular phylogenetic analyses using DNA sequence data of four loci (the ITS region, and fragments of rDNA LSU, translation elongation factor 1α and β-tubulin). The results demonstrate that Cephalotrichum, Doratomyces and Trichurus are congeneric and the genus Cephalotrichum is accepted here with Echinobotryum as a further synonym. The genera Acaulium and Fairmania, typified by A. albonigrescens and F. singularis, respectively, are distinct from Microascus and Scopulariopsis, Gamsia is distinct from Wardomyces, and Wardomycopsis is confirmed as a separate genus in the Microascaceae. Two new species of Cephalotrichum are described as C. brevistipitatum and C. hinnuleum. Nine new combinations are proposed, i.e. Acaulium acremonium, A. caviariforme, Cephalotrichum asperulum, C. columnare, C. cylindricum, C. dendrocephalum, C. gorgonifer, Gamsia columbina and Wardomyces giganteus. A neotype is designed for C. stemonitis. Lectotypes and epitypes are designated for A. acremonium, A. albonigrescens, C. gorgonifer, C. nanum and W. anomalus. Cephalotrichum cylindricum, C. microsporum, F. singularis and Gamsia columbina are also epitypified with new specimens. Descriptions of the phenotypic features and dichotomous keys for identification are provided for accepted species in the different genera.

The taxonomy of scopulariopsis-like fungi, comprising numerous human opportunistic species, has recently been reassessed with delineation of the genera Microascus, Pithoascus, Pseudoscopulariopsis, and Scopulariopsis, using morphological data and multilocus sequence analysis based on four loci (ITS, LSU, EF-1α, and TUB). In this study, the same genetic markers were used to investigate a set of clinical and environmental isolates, morphologically identified as Microascus and Scopulariopsis spp. The ingroups of the concatenated phylogenetic tree resolved 41 species clades, with isolates distributed in four main lineages corresponding to the genera Microascus, Pithoascus, Scopulariopsis, and newly established genus Fuscoannellis, typified by Scopulariopsis carbonaria. The new species Microascus chinensis, Microascus onychoides, Microascus pseudolongirostris, Pithoascus lunatus, and Scopulariopsis macurae were described. Microascus trigonosporus var. terreus and Scopulariopsis alboflavescens were found different from M. trigonosporus and Scopulariopsis brevicaulis, respectively. All the species identified in the study, except Fuscoannellis carbonaria and S. macurae, originated from clinical samples, suggesting their potential role in human disease. The use of a four marker combination was demonstrated an efficient and reliable approach to infer phylogenetic relationships among the scopulariopsis-like fungi. Yet, the only genetic marker able to discriminate all species was EF-1α, therefore proposed as a secondary barcode for the identification of these fungi.

Scopulariopsis species and their Microascus teleomorphs are cosmopolitan fungi that are uncommonly associated with invasive disease. This report describes a case of fatal disseminated Scopulariopsis brevicaulis disease in a patient with diffuse large B cell lymphoma who underwent high-dose chemotherapy followed by a matched unrelated donor stem cell transplant. This case is compared with 32 prior cases of proven invasive Scopulariopsis (Microascus) infections reported in the literature. A focus of this report is the diagnostic methods utilized which included histopathology and culture with both micromorphologic and genotypic procedures employed to confirm the species identification.

Scopulariopsis species are rarely but increasingly recognized as opportunistic pathogens in immunocompromised patients. We report on a patient suffering from cystic fibrosis who developed disseminated fungal infection due to a rare Scopulariopsis species, Microascus cirrosus , after heart and lung transplantation. Despite antifungal combination therapy with voriconazole and caspofungin, the patient died 4 weeks after transplantation. Diagnostic difficulties and optimal management of disseminated Scopulariopsis / Microascus infections are discussed.

Deep, respiratory tract and ear infections due to Microascaceae (Pseudallescheria, Scedosporium, Microascus or Scopulariopsis) were studied nationwide in Finland during 1993-2002. The data were based on 52,000 fungal cultures that represented about 50% of all such specimens in Finland and included all Finnish cases of profound immunosuppression. There were 39 cases that were re-evaluated as clinically significant, i.e., three pneumonias, two deep pedal infections and five wound infections, 11 sinusitis and 18 ear infections. The pedal infections and most pneumonias occurred in immunocompromised patients. Most cases, except the ear infections, were due to Pseudallescheria boydii. Two patients had lethal P. boydii pneumonia and a deep P. boydii infection of the foot contributed to a third lethal case. Two of the patients with lethal outcomes had received an allogeneic haematopoietic stem cell transplantation (AHSCT). Two patients with haematological malignancies were cured of deep site infections by a prolonged course of itraconazole. Wound, sinus and ear infections were cured or improved by local surgery or topical therapy. There were 0.8-1.7 cases of any type of infection per million inhabitants per year (MY) and 3.4 cases/1000 AHSCT. Mortality associated with Microascaceae in any type of patient was 0.06-0.12 MY.

Invasive fungal infections are a major cause of mortality among solid organ transplant recipients. Scopulariopsis species and their teleomorph Microascus are molds found in soil and decaying organic matter. We report here the case of a woman who underwent bilateral lung transplantation for severe emphysema. On day 25 after transplantation, endobronchial green-black lesions were detected during routine endoscopy. Endobronchial swabs, biopsies, and bronchoalveolar lavage samples were positive for Microascus cirrosus. This fungal infection developed despite voriconazole given for previous persistent invasive aspergillosis. Treatment consisted of a combination of antifungal medication (voriconazole, terbinafine, amphotericin B, and caspofungin) and endoscopic resection of necrosed bronchial mucosa. A favorable clinical outcome was achieved after 7 weeks of treatment. Seven cases of Scopulariopsis/Microascus infection have been previously described in solid organ transplant recipients. Only two survived after treatment with an antifungal combination therapy including echinocandins, posaconazole, and terbinafine. In immunocompromised patients, infection by Microascus species is a rare but life-threatening event because of innate resistance to most common antifungal drugs. Our patient was successfully cured by combined therapy including intravenous voriconazole and caspofungin, oral terbinafine, and inhaled voriconazole and amphotericin B administered for 7 weeks in association with iterative endoscopic debridement to reduce fungal inoculum.

Microascus cirrosus is very rarely the aetiological agent of onychomycosis. We report two additional cases of toenail infections caused by this fungus.

Broth microdilution checkerboard techniques based on the methodology of the Clinical and Laboratory Standards Institute (CLSI) were employed to study the triple antifungal combination of caspofungin, posaconazole, and terbinafine against 27 clinical isolates of Scopulariopsis and Microascus species. Synergy was observed for 26 isolates, whereas antagonism was observed for Scopulariopsis candida in this study.

Relationships between Scopulariopsis species and allied fungi were studied by sequencing a 350 bp gene region of the large subunit ribosomal RNA gene (LSU rDNA). In addition, a limited morphological dataset of nine characters was included in the cladistic analysis. Nineteen mitosporic strains (nine Scopulariopsis, five Wardomyces, three Doratomyces, one Trichurus and one Scedosporium species) and 21 meiosporic strains (14 Microascus, 4 Kernia and 3 Pithoascus species) were studied. The data were analysed using parsimony methods. Based on the analyses, the studied microascaceous fungi are divided to 12 molecular lineages. Most of the opportunistic human pathogenic Scopulariopsis species are placed in one clade ('Microascus manginii Clade'). Most synnematous anamorphs with Scopulariopsis-like conidia (Doratomyces and Trichurus) are placed in another clade ('Microascus albonigrescens Clade'), together with Wardomyces. Microascus sensu lato can be divided into seven clades which also incorporate all studied Pithoascus, Scopulariopsis, Wardomyces and Trichurus species and most of the Doratomyces species. Most of the Kernia teleomorphs and one Doratomyces species are placed in a different main clade, together with Pseudallescheria and Petriella. Future alternatives in the taxonomy of Microascus include splitting the genus or redefining it to include deviating taxa. More molecular data need to be obtained and considered in either case.

Several isolates of a perithecial microascaceous ascomycete having falcate ascospores and a Scopulariopsis anamorph were obtained from rotting wood in the boreal forest of Alberta, Canada. Additional isolates appeared conspecific based on anamorphic characters, but failed to produce a teleomorph. These isolates showed similarities to Microascus nidicola (type species of the genus Pithoascus) and Scopulariopsis flava. Sexual compatibility systems were investigated to establish holomorph concepts for these taxa. The teleomorph obtained in mating trials among anamorphic isolates was identical to that of self-fertile isolates. The new heterothallic species M. soppii is described. The anamorph is S. soppii. Single ascospore isolates derived from M. nidicola demonstrated homothallism and lacked an anamorph. Scopulariopsis flava (basionym Acaulium flavum) is considered a nomen dubium. Generic concepts of Pithoascus are evaluated and the genus is treated as a synonym of Microascus. Pithoascus stoveri is transferr...

Genetic relationships of twenty strains of Scedosporium, Petriella and Pseudallescheria were studied by sequencing a 370 bp part of the gene which encodes the large subunit ribosomal RNA (LSU rDNA). This study confirms and specifies earlier results from the small subunit ribosomal RNA gene (SSU rDNA). It was confirmed that the pathogenic Scedosporium prolificans is closely related to Petriella, as is Graphium tectonae. Petriella setifera, which includes clinically isolated Scedosporium strains, can be divided into several groups inside the same genus. The main part of Pseudallescheria is clearly distinguished from Petriella. Pseudallescheria is heterogeneous, however: Ps. africana is referred to Petriella and Ps. fimeti is more closely related to Microascus.