Current knowledge on the etiology and epidemiology of Scopulariopsis infections.

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.

A new species of Scopulariopsis and its synergistic effect on pathogenicity of Verticillium dahliae on cotton plants.

A new species, Scopulariopsis gossypii, was found to be present in the vascular bundles of cotton plants (Gossypium hirsutum) infected by Verticillium dahliae which is an economically important pathogen in Hangzhou, China. The fungus was only present in the diseased plants, but it never became isolated from the healthy plants. Scanning electron microscopy showed that the fungus was present in parenchyma cells enclosing vessels in dark brown vascular tissues of stems, and produced asexual conidia within the tissues. Phylogenetic analysis of combined nuclear ribosomal D1/D2 region of the 28S rDNA as well as translation elongation factor 1-alpha (TEF1) and beta-tubulin (TUB) gene showed that S. gossypii represents an undescribed species of Scopulariopsis, Microascaceae. In this study, characteristics of sexual and asexual stages of the fungus were described, illustrated and compared with similar taxa. In addition, the molecular and morphological analyses indicated that S. gossypii was a distinct species of Scopulariopsis. The pathogenicity tests proved by inoculation of wounded roots confirmed that S. gossypii was an opportunistic pathogen causing leaf interveinal chlorosis and vascular browning of cotton plants. However S. gossypii did not infect host with undamaged roots. Moreover, coinoculation with S. gossypii and V. dahliae significantly increased disease severity.

Phylogenetic Relationship and Secondary Metabolite Production of Marine Fungi Producing the Cyclodepsipeptides Scopularide A and B.

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.

Molecular taxonomy of scopulariopsis-like fungi with description of new clinical and environmental species.

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.

Especies clínicas comunes del género scopulariopsis bainier y taxas relacionados / Common clinical species of the scopulariopsis bainier genera and related taxa

Los integrantes del género Microascus y su anamorfo Scopulariopsis, incluyen especies fúngicas comunes del suelo, excrementos, ambientes internos y algunas reconocidas como oportunistas en los animales, incluyendo al hombre. La sola morfología del género no siempre permite un buen diagnóstico a pesar de su utilidad cuando se presentan los anamorfos o teleomorfos o ambos. Sin embargo, la biología molecular ha permitido el análisis de muchos aislados clínicos, contribuyendo con nuevos aportes y nuevas combinaciones taxonómicas en la literatura actual. Este trabajo, reune lo más actual sobre estos géneros comunes en salud pública, aportando claves morfofisiológicas útiles para su reconocimiento primario en los laboratorios clínicos de baja complejidad.

Members of the genus Microascus and their anamorph Scopulariopsis, include common fungal species of soil, droppings, indoor environments and some recognized opportunistic in animals, including humans. The morphology of the genus alone is not always sufficient for a proper diagnosis despite its usefulness when anamorphs or teleomorphs or both are present. However, molecular biology has allowedthe analysis of many clinical isolates, contributing with new imput and new taxonomic combinations in the current literature. This work brings together the latest on these genera in pubblic health, providing a primary key for useful morphophysiological determination in clinical laboratory of low complexity.

In Vitro Triple Combination of Antifungal Drugs against Clinical Scopulariopsis and Microascus Species.

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.

Scopulariopsis, a poorly known opportunistic fungus: spectrum of species in clinical samples and in vitro responses to antifungal drugs.

Ninety-nine isolates of clinical origin, tentatively identified as Scopulariopsis or Microascus, were morphologically and molecularly characterized by a combined analysis of the D1/D2 domains of the 28S rRNA gene and a fragment of the elongation factor 1-α gene (EF1-α) sequences. The most prevalent species was Scopulariopsis brevicaulis (49.4%), followed by Scopulariopsis gracilis (14.4%), Scopulariopsis brumptii (7.2%), Microascus cinereus (5.2%), the Scopulariopsis candida species complex (3.1%), and Microascus cirrosus (2.1%). The most common anatomic sites of isolation were the respiratory tract (61.6%), superficial tissue (19.2%), and deep tissue or fluid samples (19.2%). The antifungal susceptibilities of the isolates to eight drugs were tested in vitro, with all the drugs generally showing poor activity.

Identification of Scopulariopsis species by partial 28S rRNA gene sequence analysis.

The genus Scopulariopsis contains over 30 species of mitosporic moulds, which although usually saprophytic may also act as opportunistic pathogens in humans. They have mainly been associated with onychomycosis, and only sporadically reported as a cause of deep tissue infections or systemic disease. Identification of Scopulariopsis species still largely relies on phenotype-based methods. There is a need for a molecular diagnostic approach, that would allow to reliably discriminate between different Scopulariopsis species. The aim of this study was to apply sequence analysis of partial 28S rRNA gene for species identification of Scopulariopsis clinical isolates. Although the method employed did reveal some genetic polymorphism among Scopulariopsis isolates tested, it was not enough for species delineation. For this to be achieved, other genetic loci, within and beyond the rDNA operon, need to be investigated.

Fungal contamination of poultry litter: a public health problem.

Although numerous studies have been conducted on microbial contaminants associated with various stages related to poultry and meat products processing, only a few reported on fungal contamination of poultry litter. The goals of this study were to (1) characterize litter fungal contamination and (2) report the incidence of keratinophilic and toxigenic fungi presence. Seven fresh and 14 aged litter samples were collected from 7 poultry farms. In addition, 27 air samples of 25 litters were also collected through impaction method, and after laboratory processing and incubation of collected samples, quantitative colony-forming units (CFU/m³) and qualitative results were obtained. Twelve different fungal species were detected in fresh litter and Penicillium was the most frequent genus found (59.9%), followed by Alternaria (17.8%), Cladosporium (7.1%), and Aspergillus (5.7%). With respect to aged litter, 19 different fungal species were detected, with Penicillium sp. the most frequently isolated (42.3%), followed by Scopulariopsis sp. (38.3%), Trichosporon sp. (8.8%), and Aspergillus sp. (5.5%). A significant positive correlation was found between litter fungal contamination (CFU/g) and air fungal contamination (CFU/m³). Litter fungal quantification and species identification have important implications in the evaluation of potential adverse health risks to exposed workers and animals. Spreading of poultry litter in agricultural fields is a potential public health concern, since keratinophilic (Scopulariopsis and Fusarium genus) as well as toxigenic fungi (Aspergillus, Fusarium, and Penicillium genus) were isolated.

Invasive Scopulariopsis brevicaulis infection in an immunocompromised patient and review of prior cases caused by Scopulariopsis and Microascus species.

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.

A taxonomic and ecological overview of cheese fungi.

Cheese is made from milk by a succession of microbes (bacteria, yeasts and fungi) that determine the consistency and flavor of the cheese. Apart from the emblematic species, Penicillium camemberti and Penicillium roqueforti, cheese fungi are not well known. Here we present a taxonomic and phylogenetic overview of the most important filamentous cheese Ascomycota based on 133 isolates provided by the producers of cheese and cheese starter cultures and 97 isolates from culture collections. We checked the congruence of different gene genealogies to circumscribe cheese species and our results allow us to propose molecular targets for their identification. To study their phylogenetic affiliation, we used LSU rDNA and showed that cheese fungi are found in two classes, the Eurotiomycetes with Penicillium species (Eurotiales) and Sporendonema casei/Sphaerosporium equinum (Onygenales), and the Sordariomycetes with Scopulariopsis species (Microascales) and Fusarium domesticum (Hypocreales). Some of these fungi, such as, P. camemberti, F. domesticum, Scopulariopsis flava and S. casei, are only known from cheeses and are probably adapted to this particular habitat, which is extremely rich in protein and fat. Other cheese fungi are ubiquitous, such as, P. roqueforti, Scopulariopsis candida and Scopulariopsis fusca.

In vitro antifungal susceptibility testing of Scopulariopsis brevicaulis strains using agar diffusion method.

The genus Scopulariopsis is a common soil saprotroph and has been isolated from air, organic waste and also from plant, animal and human tissues. Scopulariopsis has mainly been associated in humans with superficial mycoses, but it has also been described as the cause of subcutaneous and invasive infections. The most common aetiological agent of infections in humans is Scopulariopsis brevicaulis. This species has been reported to be resistant in vitro to broad-spectrum antifungal agents available today. The aim of the study was to establish in vitro antifungal susceptibility of 35 S. brevicaulis strains against amphotericin B (AMB), flucytosine (FC), caspofungin (CAS), terbinafine (TER), ciclopirox (CIC), voriconazole (VOR), clotrimazole (CTR), miconazole (MCZ), econazole (ECO), ketoconazole (KET), itraconazole (ITR), and fluconazole (FLU). Antifungal susceptibility tests were evaluated by an agar diffusion method (Neo-Sensitabs, Rosco, Denmark). AMB, FC, CAS, ITR and FLU showed no antifungal activity against S. brevicaulis. TER, CIC, CTR, KET, VOR, ECO, and MCZ revealed inhibitory activity for S. brevicaulis, but it varied for each of the drugs. The best antifungal effect was observed for TER and CIC. All isolates had large inhibition zones for TER and CIC. CTR was also inhibitory for all tested S. brevicaulis isolates, but the diameters of inhibition zones were smaller than for TER and CIC. Nearly 89% isolates showed inhibition zones for KET and the mean diameter of the inhibition zone was comparable to CTR. The least antifungal activity exhibited VQR, ECO and MCZ. Because of the multiresistance of S. brevicaulis, infections due to this species may not respond to particular antifungal treatment and other therapeutic approaches should be considered, e.g., combined therapy and/or surgery.