Search

Cyp51 contribution to azole resistance has been broadly studied and characterized in Aspergillus fumigatus, whereas it remains poorly investigated in other clinically relevant species of the genus, such as those of section Nigri. In this work, we aimed to analyze the impact of cyp51genes (cyp51Aand cyp51B) on the voriconazole (VRC) response and resistance of Aspergillus nigerand Aspergillus tubingensis.

ABSTRACTCu is an essential trace element for cell growth and proliferation. However, excess of Cu accumulation leads to cellular toxicity. Thus, precise and tight regulation of Cu homeostasis processes, including transport, delivery, storage, detoxification, and efflux machineries, is required. Moreover, the maintenance of Cu homeostasis is critical for the survival and virulence of fungal pathogens. Cu homeostasis has been extensively studied in mammals, bacteria, and yeast, but it has not yet been well documented in filamentous fungi. In the present work, we investigated Cu tolerance in the filamentous fungus Fusarium oxysporumby analysing the Cu transporter coding gene crpF, previously studied in Aspergillus fumigatus. The expression studies demonstrated that crpFis upregulated in the presence of Cu and its deletion leads to severe sensitivity to low levels of CuSO4in F. oxysporum. Targeted deletion of crpFdid not significantly alter the resistance of the fungus to macrophage killing, nor its pathogenic behaviour on the tomato plants. However, the targeted deletion mutant ΔcrpFshowed increased virulence in a murine model of systemic infection compared to wild-type strain (wt).

The incidence of systemic infections by Saccharomyces cerevisiaehas increased in recent years, especially among immunocompromised patients. Amphotericin B, voriconazole or echinocandins have been used with favorable outcome against systemic infections by this fungus. However, clinical experience is limited and no in vivostudies have been conducted.

ABSTRACTThe increasing number of infections by species of Mucorales and their high mortality constitute an important concern for public health. This study aims to decipher the genetic basis of Mucor circinelloidespathogenicity, which displays virulence in a strain dependent manner. Assuming that genetic differences between strains may be linked to different pathotypes, we have conducted a study to explore genes responsible for virulence in M. circinelloidesby whole genome sequencing of the avirulent strain NRRL3631 and comparison with the virulent strain CBS277.49. This genome analysis revealed 773 truncated, discontiguous and absent genes in the NRRL3631 strain. We also examined phenotypic traits resulting in reduced heat stress tolerance, chitosan content and lower susceptibility to toxic compounds (calcofluor white and sodium dodecyl sulphate) in the virulent strain, suggesting the influence of cell wall on pathogenesis. Based on these results, we focused on studying extracellular protein-coding genes by gene deletion and further pathotype characterization of mutants in murine models of pulmonary and systemic infection. Deletion of gene ID112092, which codes for a hypothetical extracellular protein of unknown function, resulted in significant reduction of virulence.Although pathogenesis is a multifactorial process, these findings highlight the crucial role of surface and secreted proteins in M. circinelloidesvirulence and should promote further studies of other differential genes.

ABSTRACTHuman infections by coelomycetous fungi are becoming more frequent and range from superficial to systemic dissemination. Traumatic implantation of contaminated plant material is the most common cause. The typical morphological feature of these fungi is the production of asexual spores (conidia) within fruiting bodies called conidiomata. This study aimed to determine the distribution of the coelomycetes in clinical samples by a phenotypic and molecular study of a large set of isolates received from a U.S. reference mycological institution and by obtaining the in vitroantifungal susceptibility pattern of nine antifungals against a selected group of isolates. A total of 230 isolates were identified by sequencing the D1 and D2 domains of the large subunit (LSU) nuclear ribosomal RNA (nrRNA) gene and by morphological characterization. Eleven orders of the phylum Ascomycotawere identified: Pleosporales(the largest group; 66.1%), Botryosphaeriales(19.57%), Glomerellales(4.35%), Diaporthales(3.48%), Xylariales(2.17%), Hysterialesand Valsariales(0.87%), and Capnodiales, Helotiales, Hypocrealesand Magnaporthales(0.43% each). The most prevalent species were Neoscytalidium dimidiatum, Paraconiothyriumspp., Phoma herbarum, Didymella heteroderae, and Epicoccum sorghinum. The most common anatomical site of isolation was superficial tissue (66.5%), followed by the respiratory tract (17.4%). Most of the isolates tested were susceptible to the majority of antifungals, and only flucytosine showed poor antifungal activity.

Four new species ofSporidesmiumcomplex collected on plant debris in different protected areas of the Iberian Peninsula are described and illustrated.Penzigomyces basiacutusis characterized by its obclavate to rostrate conidia with a narrowly obconic-truncate basal cell and verruculose apical cell.Sporidesmium corniculatumis distinguished from other species of the genus by its occasional furcate and only transversely septate conidia andSporidesmium tunicatumby its obclavate to rostrate, verrucose conidia with an apical mucilaginous sheath tunica.Stanjehughesia obclavorostratais distinguished by its obclavate to rostrate conidia with the basal cell sometimes darker than the rest.

Cladophialophora bantianais a melanised mold with a pronounced tropism for the central nervous system, almost exclusively causing human brain abscesses.

ABSTRACTDifferent inocula of Trichoderma longibrachiatumwere tested in a murine model, and only the highest one (1 × 107CFU/animal) killed all of the mice at day 15 postinfection, with spleen and liver the most affected organs. The efficacies of amphotericin B deoxycholate, liposomal amphotericin B, voriconazole, and micafungin were evaluated in the same model, with very poor results. Our study demonstrated the low virulence but high resistance to antifungal compounds of this fungus.

ABSTRACTPenicilliumspecies are some of the most common fungi observed worldwide and have an important economic impact as well as being occasional agents of human and animal mycoses. A total of 118 isolates thought to belong to the genus Penicilliumbased on morphological features were obtained from the Fungus Testing Laboratory at the University of Texas Health Science Center in San Antonio (United States). The isolates were studied phenotypically using standard growth conditions. Molecular identification was made using two genetic markers, the internal transcribed spacer (ITS) and a fragment of the ß-tubulin gene. In order to assess phylogenetic relationships, maximum likelihood and Bayesian inference assessments were used. Antifungal susceptibility testing was performed according to CLSI document M38-A2 for nine antifungal drugs. The isolates were identified within three genera, i.e., Penicillium, Talaromyces, and Rasamsonia. The most frequent species in our study were Penicillium rubens, P. citrinum, and Talaromyces amestolkiae. The potent in vitroactivity of amphotericin B (AMB) and terbinafine (TRB) and of the echinocandins against Penicilliumand Talaromycesspecies might offer a good therapeutic alternative for the treatment of infections caused by these fungi.

ABSTRACTScopulariopsisis an emerging opportunistic fungus characterized by its high resistance to antifungal therapies. We have developed a murine model of disseminated infection in immunosuppressed animals by intravenous inoculation of Scopulariopsis brevicaulisand Scopulariopsis brumptii, the most clinically relevant species, in order to evaluate their virulence and their responses to conventional antifungal treatments. Survival and tissue burden studies showed that S. brumptiiwas more virulent than S. brevicaulis. The three drugs tested, liposomal amphotericin B, posaconazole, and voriconazole, prolonged the survival of mice infected with S. brumptii, but none showed efficacy against S. brevicaulis. The different therapies were only able to modestly reduce the fungal burden of infected tissue; however, in general, despite the high serum levels reached, they showed poor efficacy in the treatment of the infection. Unfortunately, the most effective therapy for Scopulariopsisinfections remains unresolved.

ABSTRACTAspergillus fumigatusis the main mold causing invasive fungal infection that shows high mortality rates. Therapeutic failure and the increase in drug resistance make it necessary to explore alternative treatments for this infection. We have evaluated the efficacy of amphotericin B at 0.8 mg/kg or 0.3 mg/kg of body weight combined with 40 mg/kg of posaconazole against three A. fumigatusisolates in a murine model of disseminated infection. The combination of the polyene and the azole led to a greater increase in survival and a significantly greater reduction in tissue burden than monotherapies.

ABSTRACTThe fungus Saprochaete capitatacauses opportunistic human infections, mainly in immunocompromised patients with hematological malignancies. The best therapy for this severe infection is still unknown. We evaluated the in vitrokilling activity and the in vivoefficacy of posaconazole at 5, 10, or 20 mg/kg twice a day (BID) in a murine neutropenic model of systemic infection with S. capitataby testing a set of six clinical isolates. Posaconazole showed fungistatic activity against all of the isolates tested. The different doses of the drug, especially the highest one, showed good efficacy, measured by prolonged survival, reduction of (1-3)-β-d-glucan levels in serum, tissue burden reduction, and histopathology.

ABSTRACTPeritoneal dialysis is the renal replacement modality used by ~20% of patients with end-stage kidney disease (S. McDonald, P. Clayton, and K. Hurst, p. 6.2–6.27, in ANZDATA 2012 Annual Report, 35th ed., 2012). A major complication of peritoneal dialysis is the development of peritonitis. We describe a case of Humicolasp. causing peritoneal dialysis (PD)-associated peritonitis, successfully treated with a prolonged course of antifungal therapy.

ABSTRACTCladosporiumspecies are ubiquitous, saprobic, dematiaceous fungi, only infrequently associated with human and animal opportunistic infections. We have studied a large set of Cladosporiumisolates recovered from clinical samples in the United States to ascertain the predominant species there in light of recent taxonomic changes in this genus and to determine whether some could possibly be rare potential pathogens. A total of 92 isolates were identified using phenotypic and molecular methods, which included sequence analysis of the internal transcribed spacer (ITS) region and a fragment of the large subunit (LSU) of the nuclear ribosomal DNA (rDNA), as well as fragments of the translation elongation factor 1 alpha (EF-1a) and actin (Act) genes. The most frequent species was Cladosporium halotolerans(14.8%), followed by C. tenuissimum(10.2%), C. subuliforme(5.7%), and C. pseudocladosporioides(4.5%). However, 39.8% of the isolates did not correspond to any known species and were deemed to comprise at least 17 new lineages for Cladosporium. The most frequent anatomic site of isolation was the respiratory tract (54.5%), followed by superficial (28.4%) and deep tissues and fluids (14.7%). Species of the two recently described cladosporiumlike genera Toxicocladosporiumand Penidiellaare reported for the first time from clinical samples. In vitrosusceptibility testing of 92 isolates against nine antifungal drugs showed a variety of results but high activity overall for the azoles, echinocandins, and terbinafine.

ABSTRACTExophialaspecies are capable of causing cutaneous and subcutaneous infections in immunocompromised patients. An Exophialaisolate was cultured from a biopsy specimen of a lesion on the forearm of a patient with myasthenia gravis. The patient also had lesions on the palm and distal aspects of the hand, which were successfully treated with a long-term course of itraconazole. A detailed morphological and molecular characterization of the isolate was undertaken. Phylogenetic analysis of the internal transcribed spacer region and portions of the ß-tubulin and translation elongation factor 1-alpha genes indicated that the isolate was a novel species closely related to but genetically distinct from species within the Exophiala spiniferaclade; the name Exophiala polymorphasp. nov. is proposed. Morphologically, E. polymorphamost closely resembles E. xenobioticabut it differs in possessing phialides bearing prominent, wide collarettes, and it does not produce chlamydospores.

ABSTRACTWe evaluated the combination of posaconazole with amphotericin B in vitroand in a murine model of systemic infections caused by Sporothrix brasiliensisand Sporothrix schenckiisensu stricto. In vitrodata demonstrated a synergistic effect, and although posaconazole alone was effective against sporotrichosis, efficacy in terms of survival and burden reduction was increased with the combination. This combination might be an option against disseminated sporotrichosis, especially when itraconazole or amphotericin B at optimal doses are contraindicated.

Based on a number of isolates of Myceliophthora(Chaetomiaceae, Sordariales, Ascomycota) recently isolated from soil samples collected in USA, the taxonomy of the genus was re-evaluated through phylogenetic analyses of sequences from the nuc rDNA internal transcribed spacer region and genes for the second largest subunit of RNA polymerase II and translation elongation factor 1α. Members of Myceliophthorawere split into four monophyletic clades strongly supported by molecular and phenotypic data. Such clades correspond with Myceliophthora, now restricted only to the type species of the genus Corynascus, which is re-established with five species, the new monotypic genus Crassicarponand also the new genus Thermothelomyces(comprising four species). Myceliophthora luteais mesophilic and a permanently asexual morph compared to the members of the other three mentioned genera, which also are able to sexually reproduce morphs with experimentally proven links to their asexual morphs. The asexual morph of M. luteais characterized by broadly ellipsoidal, smooth-walled conidia with a wide, truncate base. Crassicarpon thermophilumis thermophilic and heterothallic and produces spherical to cuneiform, smooth-walled conidia and cleistothecial ascomata of smooth-walled, angular cells and ascospores with a germ pore at each end. Corynascusspp. are homothallic and mesophilic and produce spherical, mostly ornamented conidia and cleistothecial ascomata with textura epidermoidea composed of ornamented wall cells, and ascospores with one germ pore at each end. Thermothelomycesspp. are thermophilic, heterothallic and characterized by similar ascomata and conidia as Corynascusspp., but its ascospores exhibit only a single germ pore. A dichotomous key to distinguish Myceliophthorafrom the other mentioned genera are provided, as well as dichotomous keys to identify the species of Corynascusand Thermothelomyces. A new species, namely Corynascus fumimontanus, characterized by verrucose ascomatal wall cells and irregularly shaped ascospores, is described and illustrated.

The mucoralean fungi are emerging causative agents of primary cutaneous infections presenting in the form of necrotizing fasciitis.

ABSTRACTAcrophialophora fusisporais an emerging opportunistic fungus capable of causing human infections. The taxonomy of the genus is not yet resolved and, in order to facilitate identification of clinical specimens, we have studied a set of clinical and environmental Acrophialophoraisolates by morphological and molecular analyses. This set included the available type strains of Acrophialophoraspecies and similar fungi, some of which were considered by various authors to be synonyms of A. fusispora. Sequence analysis of the large subunit (LSU) and internal transcribed spacer (ITS) regions of the nuclear ribosomal DNA and a fragment of the ß-tubulin (Tub) gene revealed that Acrophialophorabelongs in the family Chaetomiaceaeand comprises three different species, i.e., A. fusispora, Acrophialophora levis, and Acrophialophora seudatica; the latter was previously included in the genus Ampullifera. The most prevalent species among clinical isolates was A. levis(72.7%), followed by A. fusispora(27.3%), both of which were isolated mostly from respiratory specimens (72.7%), as well as subcutaneous and corneal tissue samples. In general, of the eight antifungal drugs tested, voriconazole had the greatest in vitroactivity, while all other agents showed poor in vitroactivity against these fungi.

There are very few drug delivery systems that target key organs via the oral route, as oral delivery advances normally address gastrointestinal drug dissolution, permeation, and stability. Here we introduce a nanomedicine in which nanoparticles, while also protecting the drug from gastric degradation, are taken up by the gastrointestinal epithelia and transported to the lung, liver, and spleen, thus selectively enhancing drug bioavailability in these target organs and diminishing kidney exposure (relevant to nephrotoxic drugs). Our work demonstrates, for the first time, that oral particle uptake and translocation to specific organs may be used to achieve a beneficial therapeutic response. We have illustrated this using amphotericin B, a nephrotoxic drug encapsulated within N-palmitoyl-N-methyl-N,N-dimethyl-N,N,N-trimethyl-6-O-glycol chitosan (GCPQ) nanoparticles, and have evidenced our approach in three separate disease states (visceral leishmaniasis, candidiasis, and aspergillosis) using industry standard models of the disease in small animals. The oral bioavailability of AmB-GCPQ nanoparticles is 24%. In all disease models, AmB-GCPQ nanoparticles show comparable efficacy to parenteral liposomal AmB (AmBisome). Our work thus paves the way for others to use nanoparticles to achieve a specific targeted delivery of drug to key organs via the oral route. This is especially important for drugs with a narrow therapeutic index.

ABSTRACTRecent changes in the Fungal Code of Nomenclature and developments in molecular phylogeny are about to lead to dramatic changes in the naming of medically important molds and yeasts. In this article, we present a widely supported and simple proposal to prevent unnecessary nomenclatural instability.

ABSTRACTThe use of molecular identification techniques has revealed an increasing number of new species within Aspergillussection Terrei. We phenotyped a set of 26 clinical isolates that showed genetic differences from Aspergillus terreussensu strictoby analyzing sequences from PCR-amplified ß-tubulin and calmodulin genes and the internal transcribed spacer region. Since the isolates were phylogenetically and morphologically different from all of the members of Aspergillussection Terrei, they are described here as a new species, Aspergilluscitrinoterreus, so named because it produces a diffusible yellowish pigment in agar. A. citrinoterreusisolates were significantly more susceptible to itraconazole, voriconazole, and posaconazole than A. terreussensu strictoisolates were; in contrast, the amphotericin B MICs for both species were high. A. citrinoterreuswas found in clinical samples from patients with proven or probable invasive aspergillosis and colonized patients, none of whom had hematological malignancies as predisposing conditions. However, they did have other underlying conditions such as chronic obstructive pulmonary disease, cirrhosis, and cancer or had received a solid organ transplants and presented not only with invasive pulmonary aspergillosis but also with mediastinitis. A. citrinoterreusisolates were detected for the first time in 2002. In all cases of invasive aspergillosis, A. citrinoterreuswas found to be a copathogen, mostly with A. fumigatus.

Candida guilliermondiihas been recognized as an emerging pathogen showing a decreased susceptibility to fluconazole and considerably high echinocandin MICs.

ABSTRACTA case of fungal necrotizing fasciitis that appeared in an immunocompetent Mexican woman after a car accident is described. The patient did not respond to antifungal treatment and died 4 days later. The fungus was molecularly identified as a new species of Apophysomyces, namely, Apophysomyces mexicanus.

ABSTRACTOchroconisis a dematiaceous fungus able to infect immunocompetent people. Recently, the taxonomy of the genus has been reevaluated, and the most relevant species, Ochroconis gallopava, was transferred to the new genus Verruconis. Due to the important clinical implications of these fungi and based on the recent classification, it was of interest to know the spectra of Ochroconisand Verruconisspecies in clinical samples received in a reference laboratory in the United States. A set of 51 isolates was identified morphologically and molecularly based on sequence analyses of the nuclear ribosomal RNA (nrRNA), actin, and ß-tubulin genes. Verruconis gallopavawas the most common species (68.6%), followed by Ochroconis mirabilis(21.5%). One isolate of Ochroconis cordanaewas found, being reported for the first time in a clinical setting. The most common anatomical site of isolation was the lower respiratory tract (58.8%), followed by superficial and deep tissues at similar frequencies (21.6 and 19.6%, respectively). Interestingly, three new species were found, which are Ochroconis olivaceaand Ochroconis ramosafrom clinical specimens and Ochroconis icarusof an environmental origin. The in vitroantifungal susceptibilities of eight antifungal drugs against the Ochroconisisolates revealed that terbinafine and micafungin were the most active drugs.

Two interesting fungi belonging to the genus Cordanahave been isolated recently in Spain from plant debris. Both are proposed here as new species, described and illustrated. Cordana mercadianasp. nov. produces 0–1-septate conidia, with a prominent basal scar. Cordana verruculosasp. nov. differs from the other species of the genus by its unique combination of aseptate, verruculose and small conidia. Both species are compared morphologically with other species of Cordanaand their identities supported by the analysis of rDNA sequences. LSU sequence analysis revealed the congeneric relationship of Cordanaand Pseudobotrytis; the members of both genera are in a well supported monophyletic lineage that appears to be related to the Coniochaetales but remains incertae sedis within the Sordariomycetes. To establish nomenclatural stability of the genus Cordana, an isolate of C. pauciseptatais designed here as epitype and the two species of Pseudobotrytisare transferred to Cordana. A dichotomous key is provided to identify the currently accepted species of Cordana.

ABSTRACTA set of 73 isolates of the emerging fungus Trichodermaisolated from human and animal clinical specimens were characterized morphologically and molecularly using a multilocus sequence analysis that included the internal transcribed spacer (ITS) regions of the nuclear ribosomal DNA and fragments of the translation elongation factor 1 alpha (Tef1), endochitinase CHI18-5 (Chi18-5), and actin 1 (Act1) genes. The most frequent species was Trichoderma longibrachiatum(26%), followed by Trichoderma citrinoviride(18%), the Hypocrea lixii/Trichoderma harzianumspecies complex (15%), the newly described species Trichoderma bissettii(12%), and Trichoderma orientale(11%). The most common anatomical sites of isolation in human clinical specimens were the respiratory tract (40%), followed by deep tissue (30%) and superficial tissues (26%), while all the animal-associated isolates were obtained from superficial tissue samples. Susceptibilities of the isolates to eight antifungal drugs in vitroshowed mostly high MICs, except for voriconazole and the echinocandins.

To ensure a stable platform for fungal taxonomy, it is of paramount importance that the genetic application of generic names be based on their DNA sequence data, and wherever possible, not morphology or ecology alone. To facilitate this process, a new database, accessible at https://doi.org/www.GeneraofFungi.org(GoF) was established, which will allow deposition of metadata linked to holo-, lecto-, neo- or epitype specimens, cultures and DNA sequence data of the type species of genera. Although there are presently more than 18 000 fungal genera described, we aim to initially focus on the subset of names that have been placed on the “Without-prejudice List of Protected Generic Names of Fungi” (see IMA Fungus4(2): 381–443, 2013). To enable the global mycological community to keep track of typification events and avoid duplication, special MycoBank Typification identfiers (MBT) will be issued upon deposit of metadata in MycoBank. MycoBank is linked to GoF, thus deposited metadata of generic type species will be displayed in GoF (and vice versa), but will also be linked to Index Fungorum (IF) and the curated RefSeq Targeted Loci (RTL) database in GenBank at the National Center for Biotechnology Information (NCBI). This initial paper focuses on eight genera of appendaged coelomycetes, the type species of which are neo- or epitypified here: Bartalinia (Bartalinia robillardoides; Amphisphaeriaceae, Xylariales), Chaetospermum (Chaetospermum chaetosporum, incertae sedis, Sebacinales), Coniella (Coniella fragariae, Schizoparmaceae, Diaporthales), Crinitospora (Crinitospora pulchra, Melanconidaceae, Diaporthales), Eleutheromyces (Eleutheromyces subulatus, Helotiales), Kellermania (Kellermania yuccigena, Planistromataceae, Botryosphaeriales), Mastigosporium (Mastigosporium album, Helotiales), and Mycotribulus (Mycotribulus mirabilis, Agaricales).Authors interested in contributing accounts of individual genera to larger multi-authored papers to be published in IMA Fungus, should contact the associate editors listed below for the major groups of fungi on the List of Protected Generic Names for Fungi.

ABSTRACTIt has been argued that the in vitroactivity of caspofungin (CSP) is not a good predictor of the outcome of echinocandin treatment in vivo. We evaluated the in vitroactivity of CSP and the presence of FKSmutations in the hot spot 1 (HS1) region of the FKS1and FKS2genes in 17 Candida glabratastrains with a wide range of MICs. The efficacy of CSP against systemic infections from each of the 17 strains was evaluated in a murine model. No HS1 mutations were found in the eight strains showing MICs for CSP of ≤0.5 μg/ml, but they were present in eight of the nine strains with MICs of ≥1 μg/ml, i.e., three in the FKS1gene and five in the FKS2gene. CSP was effective for treating mice infected with strains with MICs of ≤0.5 μg/ml, showed variable efficacy in animals challenged with strains with MICs of 1 μg/ml, and did not work in those with strains with MICs of >1 μg/ml. In addition, mutations, including one reported for the first time, were found outside the HS1 region in the FKS2gene of six strains with different MICs, but their presence did not influence drug efficacy. The in vitroactivity of CSP was compared with that of another echinocandin, anidulafungin, suggesting that the MICs of both drugs, as well as mutations in the HS1 regions of the FKS1and FKS2genes, are predictive of outcome.

Acremoniumis one of the largest and most complex genera of hyphomycetes. Its taxonomy is not yet resolved and the phylogenetic value of the most distinctive morphological features is unknown. The species of this genus produce conidia in chains or form slimy heads or both. We have studied a set of clinical and environmental Acremoniumstrains morphologically characterized by producing mostly catenate, elongate, more or less fusiform conidia. Based on phenotypic data and in the analysis of the sequences of the ITS region, the D1/D2 domains of the 28S rRNA gene and a fragment of the β-tubulin gene, three new species morphologically similar to Acremonium fusidioidesand belonging to the family Bionectriaceae (Hypocreales) are described and illustrated; they are Acremonium pilosum, Acremonium parvumand Acremonium citrinum. The first species produces two kinds of conidia: clavate with smooth walls and globose with abundant filiform projections. Acremonium parvumis characterized by slow growth and pyriform or slightly lanceolate conidia with an elongate and truncate base. Acremonium citrinumproduces a diffusible yellow pigment and obovoid conidia.

ABSTRACTNinety-nine isolates of clinical origin, tentatively identified as Scopulariopsisor 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-a gene (EF1-a) 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 candidaspecies 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.

ABSTRACTTriadelphia pulvinatais a rare dematiaceous fungus found in soil. We report the first case of invasive disease in a patient with acute myeloid leukemia who had a bloodstream infection with possibly both lung and brain involvement. Identification was by combined phenotypic features and fungal ribosomal DNA sequence analysis.

ABSTRACTWe evaluated and compared the efficacies of different antifungal drugs against Sarocladium kiliense(formerly Acremonium kiliense), a clinically relevant opportunistic fungus, in a murine model of systemic infection. Three clinical strains of this fungus were tested, and the therapy administered was as follows: posaconazole at 20 mg/kg of body weight (twice daily), voriconazole at 40 mg/kg, anidulafungin at 10 mg/kg, or amphotericin B at 0.8 mg/kg. The efficacy was evaluated by prolonged animal survival, tissue burden reduction, and (1→3)-β-d-glucan serum levels. In general, the four antifungal drugs showed high MICs and poor in vitroactivity. The efficacy of the different treatments was only modest, since survival rates were never higher than 40% and no drug was able to reduce fungal load in all the organs for the three strains tested. Posaconazole, in spite of its high MICs (≥16 μg/ml), showed the highest efficacy. The (1→3)-β-d-glucan serum levels were equally reduced by all drugs evaluated.

In molecular studies involving numerous clinical isolates of the genera Acremonium, Phialemoniumand Lecytophorasome of them could not be identified. To clarify the phylogenetic relationships among these fungi and other related taxa, we performed a polyphasic study based on a detailed morphological study and on the analysis of sequences of four loci: the internal transcribed spacer regions, the D1/D2 domains of the 28S rRNA, actin and β-tubulin genes. The combination of the resulting data let us propose the new genus Phialemoniopsisto accommodate the opportunistic fungi Phialemonium curvatumand Sarcopodium oculorumand two new species, Phialemoniopsis cornearisand Phialemoniopsis pluriloculosa. The taxonomy of Phialemoniopsishas not been completely resolved, however, remaining incertae sedis within the Sordariomycetes. In addition, the new species Lecythophora luteorubra, Lecythophora cateniformisand Phialemonium globosumare described and the species Acremonium atrogriseumand Taifanglania inflataare transferred to the genus Phialemonium. Lecythophoraand Phialemoniumare currently monophyletic genera of the families Coniochaetaceae (Coniochaetales) and Cephalothecaceae (Sordariales) respectively, according to our results. Tables summarizing key morphological features to distinguish the current species of Lecythophora, Phialemoniumand Phialemoniopsisare provided.

ABSTRACTThe in vitroantifungal susceptibility of 77 isolates belonging to different clinically relevant species of Aspergillussection Flavi, including those of different phylogenetic clades of A. flavus, was tested for nine antifungal agents using a microdilution reference method (CLSI, M38-A2). Terbinafine and the echinocandins demonstrated lower MICs/MECs for all species evaluated, followed by posaconazole. Amphotericin B showed MICs ≥ 2 μg/ml for 38 (49.4%) of the 77 isolates tested.

Purpureocilliumis a genus recently proposed to accommodate Paecilomyces lilacinus, a well studied species that has biotechnological properties and an ability to cause human infections. Since contradictory data have been reported on the intraspecific genetic variability of P. lilacinum, we have carried out a polyphasic study of a set of clinical and environmental isolates of this species. Detailed morphological examination and sequence analysis of four different loci, including the ribosomal internal transcribed spacer, the domains D1 and D2 of the 28S rDNA, EF-1aand the largest subunit of RNA polymerase II (rpb1), showed that P. lilacinumformed a well supported phylogenetic clade with low intraspecific variability. The new species Purpureocillium lavendulum, which has vinaceous colonies similar to those of P. lilacinum, is proposed. It is characterized by the lack of growth at 35 C, the production of a yellow diffusible pigment and by subglobose or limoniform conidia.

ABSTRACTWe have evaluated the in vitroactivity of voriconazole against 61 strains of Aspergillus fumigatusby using broth microdilution, disk diffusion, and minimal fungicidal concentration procedures. We observed an excellent correlation between the results obtained with the three methods. Five percent of the strains showed MICs greater than or equal to the epidemiological cutoff value (ECV; 1 μg/ml). To assess whether MICs were predictive of in vivooutcome, we tested the efficacy of voriconazole at 25 mg/kg of body weight daily in an immunosuppressed murine model of disseminated infection using 10 strains representing various patterns of susceptibility to the drug as determined by the in vitrostudy. Voriconazole prolonged survival and reduced fungal load in the kidneys and brain in those mice infected with strains with MICs of ≤0.25 μg/ml, while in mice infected with strains with MICs of 0.5 to 2 μg/ml, the efficacy was varied and strain dependent and in mice infected with the strain with a MIC of 4 μg/ml, the antifungal did not show efficacy. Voriconazole reduced galactomannan antigenemia against practically all strains with a MIC of <4 μg/ml. Our results demonstrate that some relationship exists between voriconazole MICs and in vivoefficacy; however, further studies testing additional strains are needed to better ascertain which MIC values can predict clinical outcome.

In a survey on the diversity of microfungi in Spanish soils, two new species of Acremoniumwere found. Both species were characterized as having more or less erect, mostly branched conidiophores bearing whorls of acicular phialides. In addition, one of these species, Acremonium asperulatum, produced abundant chlamydospores and globose rough-walled conidia. The other species, Acremonium variecolor, produced a brownish diffusible pigment and smooth-walled, subglobose conidia with apiculate base; sessile conidia inserted directly on vegetative hyphae also were present. The analysis of the sequences of the ITS region, the D1/D2 domains of the 28S rRNA gene and a fragment of the actin gene revealed relationships of both species with members of the Bionectriaceae (Hypocreales). Genetic differences were observed with morphologically similar species.

Saksenaea erythrosporais a filamentous fungus belonging to the order Mucorales. Cases of cutaneous mucormycosis caused by Saksenaeaspp. have previously been reported in immunocompetent and immunosuppressed people. A premature, 1-day-old bull calf from Texas with numerous plaque-like and ulcerative lesions in the skin was found at necropsy to have multiple areas of mycotic dermatitis and abomasitis. Fungal culture of the skin followed by morphological characterization and genetic analysis identified the etiologic agent as S. erythrospora.

ABSTRACTApophysomyces variabilisis an emerging fungal pathogen that can cause significant infections in immunocompetent patients. We report a case of A. variabilisinvasive wound infection in a 21-year-old male after a self-inflicted burn injury.

ABSTRACTCandida rugosais a poorly known fungal species occasionally involved in human infections. A molecular analysis of the sequences of the D1/D2 domains and the internal transcribed spacer (ITS) region of the ribosomal genes of 24 clinical isolates phenotypically identified as C. rugosademonstrated that only 10 (41.6%) isolates belonged to that species. The other isolates were identified as Candida pararugosa(41.6%) and Candida pseudorugosa(8.3%). The remaining two isolates, from human and equine infections, respectively, were clearly different from the others and represent a new species proposed here as Candida neorugosa. The closest species by D1/D2 sequences was the type strain of C. rugosa, with only 92.3% similarity. C. neorugosacan also be differentiated from all other species of the C. rugosacomplex by phenotypic features. The eight antifungal drugs tested showed high in vitroactivity against the 24 isolates included in the study.

ABSTRACTExserohilumis an agent of human and animal mycoses. Although classification has been based on a few subtle morphological differences, three species of clinical interest have been traditionally accepted. In this study, by using a multigene sequence analysis, we have demonstrated that Exserohilum longirostratumand E. mcginnisiiare probable synonyms of E. rostratum. The isolates tested were mainly from the nasal region. Antifungal susceptibility testing demonstrated high activity of the eight agents tested against this fungus.

ABSTRACTThe first case of cavitary pulmonary disease caused by Purpureocillium lilacinumis described. The isolate showed atypical microscopic characteristics similar to Acremoniumand Fusariumspp., which necessitated molecular identification by sequencing of multiple conserved loci. The patient responded to voriconazole, reinforcing its therapeutic efficacy for P. lilacinuminfections.