Identification and characterization of compounds from Chrysosporium multifidum, a fungus with moderate antimicrobial activity isolated from Hermetia illucens gut microbiota.

The gut microbiota of insects is composed of a wide range of microorganisms which produce bioactive compounds that protect their host from pathogenic attack. In the present study, we isolate and identify the fungus Chrysosporium multifidum from the gut of Hermetia illucens larvae. Extract from C. multifidum culture broth supernatant showed moderate activity against a strain of methicillin-resistant Staphylococcus aureus (MRSA). Bioguided isolation of the extract resulted in the characterization of six α-pyrone derivatives (1-6) and one diketopiperazine (7). Of these compounds, 5,6-dihydro-4-methoxy-6-(1-oxopentyl)-2H-pyran-2-one (4) showed the greatest activity (IC50 = 11.4 ± 0.7 µg/mL and MIC = 62.5 µg/mL) against MRSA.

Disseminated Emergomycosis in a Person with HIV Infection, Uganda.

We describe emergomycosis in a patient in Uganda with HIV infection. We tested a formalin-fixed, paraffin-embedded skin biopsy to identify Emergomyces pasteurianus or a closely related pathogen by sequencing broad-range fungal PCR amplicons. Results suggest that emergomycosis is more widespread and genetically diverse than previously documented. PCR on tissue blocks may help clarify emergomycosis epidemiology.

Disseminated cutaneous chrysosporium infection.

Cutaneous chrysosporium infection is extremely rare and underdiagnosed. We present an immunocompromised patient who presented with recurrent cutaneous abscesses. Histopathology of the abscess showed thick-walled conidia and septate fungal hyphae within the subcutis and fungal culture grew Chrysosporium species.

Endangered Zoonotic Fungal Species from Chicken (Gallus gallus domestics).

Lophophyton gallinae (Microsporum gallinae) is a zoophilic fungus that causes ringworm in chickens and related species, and occasionally in humans. There are 45 human cases worldwide including a Japanese case from Okinawa in 2009. After the finding of the human L. gallinae case, 793 chickens in Japan, including 293 from the mainland and 500 from the Nansei Island areas, were investigated to determine the prevalence of dermatophytes and their related fungal species. The survey was carried out from December 2008 to March 2013. Various dermatophytes and related fungal species were isolated from the studied chickens, with a prevalence of 24.6%. In total, 224 dermatophytes and related species were isolated in the survey. The most commonly isolated species included, in descending order of frequency, Arthroderma multifidum, Aphanoascus terreus, and Chrysosporium spp. Ar. multifidum and Ap. terreus have no record of pathogenicity, and the present isolates of Chrysosporium spp. were not matched to pathogenic Chrysosporium spp. based on the ITS rDNA sequences. Interestingly, an L. gallinae isolate was detected in a male 10-month-old shamo (fighting cock) from the main island. Furthermore, one strain of Arthroderma simii was also isolated as the second record in Japan following that from an imported chimpanzee. Although L. gallinae and Ar. simii are likely to be endemic in our country, the transmission of dermatophytosis from chickens to humans is unlikely to occur because of the reduced chances for citizens to come in contact with chickens due to various factors.

Adiaspiromycosis in a wild European rabbit, and a review of the literature.

Adiaspiromycosis is a mycotic infection caused by thermally dimorphic fungi classified as Emmonsia parva and E. crescens (formerly Chrysosporium spp.) until recently, when new classifications were proposed. We document the pathologic findings in a severe case of adiaspiromycosis, with lymph node involvement, in a wild European rabbit ( Oryctolagus cuniculus). The rabbit exhibited granulomatous pneumonia with tracheobronchial lymph node enlargement. Histopathologically, the lung was expanded by myriad, densely cellular, heterophilic and granulomatous foci, surrounding bi- to trilaminar adiaspores. Adiaspore density was considered to be similar in all lung lobes. In the left caudal lung lobe, 80 adiaspores were counted in a 50-mm2 area using digital image analysis. The mean and median adiaspore diameters were 240 ± 52 µm and 255 µm, respectively. Tracheobronchial lymph nodes exhibited moderate numbers of similar adiaspores. PCR amplification of DNA extracted from microdissected adiaspores failed to identify Emmonsia spp.-specific DNA. These data suggest that adiaspiromycosis may result in severe granulomatous pneumonia in wild European rabbits. Although confirmation of the etiologic agent by PCR using DNA extracted from formalin-fixed tissue is not always successful, digital image analysis can be used to aid accurate assessment of adiaspore density and morphology.

Pulmonary adiaspiromycosis in the Eurasian beaver (Castor fiber) inhabiting Poland.

Adiaspiromycosis is a rare fungal infection caused by saprophytic fungi Emmonsia spp. (type Ascomycota) occurring especially in small free-living mammals. The aim of this study was to evaluate the occurrence of histopathological lesions asscociated with adiaspiromycosis in the Eurasian beaver inhabiting Poland. In order to evaluate the presence of natural adiaspiromycosis we systematically investigated beaver populations from north-eastern Poland for adiaspores in the lungs. This study reveals for the first time the presence of pulmonary adiaspiromycosis of Eurasian beaver in Poland. As far as we know, there is no published data regarding pulmonary adiaspiromycosis in human patients in Poland.

Pulmonary adiaspiromycosis in critically endangered northern hairy-nosed wombats (Lasiorhinus krefftii).

BACKGROUND: The northern hairy-nosed wombat (Lasiorhinus krefftii) is critically endangered, with only 200 individuals remaining in the wild. Individuals are rarely available for detailed pathological assessment and identification of disease threats to individuals is critically important to species conservation. CASE REPORT: Two male northern hairy-nosed wombats, part of the Richard Underwood Nature Refuge population, were presented for necropsy, 5 months apart. They were found to have succumbed to adiaspiromycosis caused by the fungus Emmonsia parva. Pathological presentations were of severe pulmonary oedema and fibrosis, and pleuritis, respectively. Characteristic fungal adiaspores were noted on histopathological examination. The wombats had concurrent variably severe ectoparasite and endoparasite burdens. CONCLUSION: These are the first reported cases of adiaspiromycosis in northern hairy-nosed wombats and the organism was associated with significant pathological changes. The rarity and the logistical challenges of presenting northern hairy-nosed wombats for pathological assessment are a challenge to identifying disease threats in this critically endangered species.

In Vitro Antifungal Susceptibility of Yeast and Mold Phases of Isolates of Dimorphic Fungal Pathogen Emergomyces africanus (Formerly Emmonsia sp.) from HIV-Infected South African Patients.

Disseminated emmonsiosis is an important AIDS-related mycosis in South Africa that is caused by Emergomycesafricanus, a newly described and renamed dimorphic fungal pathogen. In vitro antifungal susceptibility data can guide management. Identification of invasive clinical isolates was confirmed phenotypically and by sequencing of the internal transcribed spacer region. Yeast and mold phase MICs of fluconazole, voriconazole, itraconazole, posaconazole, caspofungin, anidulafungin, micafungin, and flucytosine were determined with custom-made frozen broth microdilution (BMD) panels in accordance with Clinical and Laboratory Standards Institute recommendations. MICs of amphotericin B, itraconazole, posaconazole, and voriconazole were determined by Etest. Fifty unique E. africanus isolates were tested. The yeast and mold phase geometric mean (GM) BMD and Etest MICs of itraconazole were 0.01 mg/liter. The voriconazole and posaconazole GM BMD MICs were 0.01 mg/liter for both phases, while the GM Etest MICs were 0.001 and 0.002 mg/liter, respectively. The fluconazole GM BMD MICs were 0.18 mg/liter for both phases. The GM Etest MICs of amphotericin B, for the yeast and mold phases were 0.03 and 0.01 mg/liter. The echinocandins and flucytosine had very limited in vitro activity. Treatment and outcome data were available for 37 patients; in a multivariable model including MIC data, only isolation from blood (odds ratio [OR], 8.6; 95% confidence interval [CI], 1.3 to 54.4; P = 0.02) or bone marrow (OR, 12.1; 95% CI, 1.2 to 120.2; P = 0.03) (versus skin biopsy) was associated with death. In vitro susceptibility data support the management of disseminated emmonsiosis with amphotericin B, followed by itraconazole, voriconazole, or posaconazole. Fluconazole was a relatively less potent agent.

Biodegradation of feather waste keratin by a keratinolytic soil fungus of the genus Chrysosporium and statistical optimization of feather mass loss.

This paper assesses the ability of strains of Aphanoascus fulvescens and Chrysosporium articulatum isolated from soil (phaesol) to degrade native feather keratin. Strains were identified based on phenotypic traits and nucleotide sequencing. Response Surface Methodology was used to optimize cultivation conditions exhibiting the highest keratinolytic activity. The experiments were based on Box-Behnken designs for the loss of substrate mass (chicken feathers). While substrate mass loss is an "economic coefficient" that reliably indicates feather keratin degradation, it has not been studied before. Stationary liquid cultures of five selected strains were conducted in laboratory conditions at 28 °C using poultry feathers (1 g) as the sole source of carbon, nitrogen and energy. Enzymatic activities, keratin mineralization products and substrate mass loss were determined periodically. The mineralization of keratin proteins by strains yielded a high number of ammonium ions alkalinizing the medium. Increased ammonium ions inhibited the activity of caseinian protease and keratinase. A decrease in the concentration of these ions induced proteolytic enzymes, chiefly the activity of keratinase, at the end of fungal cultivation. Keratinase activity was related to protein- and peptide release and that of caseinian protease to sulfate ions. The highest loss of substrate mass in comparison to the reference strain CBS104.62 (35.4%) was recorded for Aphanoascus fulvescens B21/4-5 (65.9%). Based on a Box-Behnken design, the maximum loss of substrate mass for the Aphanoascus fulvescens strain (71.08%) can be achieved at pH 7.58 and temperature 28.7 °C.

Snake fungal disease caused by Ophidiomyces ophiodiicola in a free-ranging mud snake (Farancia abacura).

Snake fungal disease is an emerging infectious disease caused by the fungus Ophidiomyces ophiodiicola leading to severe dermatitis and facial disfiguration in numerous free-ranging and captive snakes. A free-ranging mud snake (Farancia abacura) from Bulloch County, Georgia, was presented for autopsy because of facial swelling and emaciation. Extensive ulceration of the skin, which was especially severe on the head, and retained shed were noted on external examination. Microscopic examination revealed severe heterophilic dermatitis with intralesional fungal hyphae and arthroconidia consistent with O. ophiodiicola A skin sample incubated on Sabouraud dextrose agar yielded a white-to-tan powdery fungal culture that was confirmed to be O. ophiodiicola by polymerase chain reaction and sequence analysis. Heavy infestation with adult tapeworms (Ophiotaenia faranciae) was present within the intestine. Various bacterial and fungal species, interpreted to either be secondary invaders or postmortem contaminants, were associated with oral lesions. Although the role of these other organisms in the overall health of this individual is not known, factors such as concurrent infections or immunosuppression should be considered in order to better understand the overall manifestation of snake fungal disease, which remains poorly characterized in its host range and geographic distribution.

Dermatomycosis in three central bearded dragons (Pogona vitticeps) associated with Nannizziopsis chlamydospora.

Chronic dermatomycosis was identified in 3 central bearded dragons (Pogona vitticeps), held as companion animals by the same owner. Clinical signs of dermatomycosis included subcutaneous masses as well as crusty, erosive, and ulcerative skin lesions. The facial region was affected in 2 of the 3 cases. Masses were surgically excised, and histology confirmed necrotizing and granulomatous inflammatory processes associated with fungal hyphae. Two of the bearded dragons were euthanized because of their deteriorating condition. In both cases, postmortem histology confirmed systemic fungal infections despite treatment of 1 animal with itraconazole. In the third bearded dragon, therapy with voriconazole at 10 mg/kg was initially effective, but mycotic lesions reappeared 15 months later. Nannizziopsis chlamydospora was identified by PCR and subsequent DNA sequencing in 2 of these cases.

Isolation and Characterization of an Unknown Chrysosporium sp. Producing Subcutaneous Mycosis in an Immunocompromised Patient.

We report a case of subcutaneous fungal abscess over the great toe caused by a keratinophilic fungus, an unknown Chrysosporium sp., in a 60-year-old diabetic female who was treated successfully with oral fluconazole. The fungus was isolated from aspirated pus, and septate hyphae were seen in fine needle aspiration cytology. Ovoid- to club-shaped hyaline one-celled conidia (aleuriconidia) with broad truncated bases were seen, and sequencing of ITS1-5.8S-ITS2 rDNA revealed belonging to the order Onygenales and most closely related to Chrysosporium spp. isolated from a fowl. Of the 65 species within the genus Chrysosporium, very few have been reported as pathogenic.

Disseminated Emmonsia pasteuriana infection in India: a case report and a review.

We report here the first case of disseminated Emmonsia pasteuriana infection in a patient with AIDS in India. The patient presented with weight loss, dyspnoea, left-sided chest pain and multiple non-tender skin lesions over face and body for 3 months. Disseminated emmonsiosis was diagnosed on microscopic examination and fungal culture of skin biopsy and needle aspirate of lung consolidation. It was confirmed by sequencing internal transcribed spacer region of rDNA, beta tubulin, actin, and intein PRP8. The patient responded to amphotericin B and itraconazole therapy.

Disseminated infection caused by Emmonsia pasteuriana in a renal transplant recipient.

Emmonsia pasteuriana is a thermally dimorphic fungus identified in very few human cases. Here, we report a case of a 43-year-old male renal transplant patient from China presenting with multiple painful skin eruptions on his head, nose and left thigh, later accompanied by respiratory failure. Histopathology of the biopsy collected from the left thigh upper ulcer and occipital nodule both demonstrated chronic inflammation with granuloma formation and yeast-like elements. Emmonsia pasteuriana was cultured from two biopsy specimens and their identity was confirmed by sequencing of the rDNA internal transcribed spacer. The patient in intensive care showed marked clinical improvement with antifungal treatment.

Clinical Characteristics, Diagnosis, Management, and Outcomes of Disseminated Emmonsiosis: A Retrospective Case Series.

BACKGROUND: We describe the geographic distribution, clinical characteristics, and management of patients with disease caused by Emmonsia sp., a novel dimorphic fungal pathogen recently described in South Africa. METHODS: We performed a multicenter, retrospective chart review of laboratory-confirmed cases of emmonsiosis diagnosed across South Africa from January 2008 through February 2015. RESULTS: Fifty-four patients were diagnosed in 5/9 provinces. Fifty-one patients (94%) were human immunodeficiency virus coinfected (median CD4 count 16 cells/µL [interquartile range, 6-40]). In 12 (24%) of these, antiretroviral therapy had been initiated in the preceding 2 months. All patients had disseminated disease, most commonly involving skin (n = 50/52, 96%) and lung (n = 42/48, 88%). Yeasts were visualized on histopathologic examination of skin (n = 34/37), respiratory tissue (n = 2/4), brain (n = 1/1), liver (n = 1/2), and bone marrow (n = 1/15). Emmonsia sp. was cultured from skin biopsy (n = 20/28), mycobacterial/fungal and aerobic blood culture (n = 15/25 and n = 9/37, respectively), bone marrow (n = 12/14), lung (n = 1/1), lymph node (n = 1/1), and brain (n = 1/1). Twenty-four of 34 patients (71%) treated with amphotericin B deoxycholate, 4/12 (33%) treated with a triazole alone, and none of 8 (0%) who received no antifungals survived. Twenty-six patients (48%) died, half undiagnosed. CONCLUSIONS: Disseminated emmonsiosis is more widespread in South Africa and carries a higher case fatality rate than previously appreciated. Cutaneous involvement is near universal, and skin biopsy can be used to diagnose the majority of patients.

TaqMan real-time polymerase chain reaction for detection of Ophidiomyces ophiodiicola, the fungus associated with snake fungal disease.

BACKGROUND: Fungal skin infections associated with Ophidiomyces ophiodiicola, a member of the Chrysosporium anamorph of Nannizziopsis vriesii (CANV) complex, have been linked to an increasing number of cases of snake fungal disease (SFD) in captive snakes around the world and in wild snake populations in eastern North America. The emergence of SFD in both captive and wild situations has led to an increased need for tools to better diagnose and study the disease. RESULTS: We developed two TaqMan real-time polymerase chain reaction (PCR) assays to rapidly detect O. ophiodiicola in clinical samples. One assay targets the internal transcribed spacer region (ITS) of the fungal genome while the other targets the more variable intergenic spacer region (IGS). The PCR assays were qualified using skin samples collected from 50 snakes for which O. ophiodiicola had been previously detected by culture, 20 snakes with gross skin lesions suggestive of SFD but which were culture-negative for O. ophiodiicola, and 16 snakes with no clinical signs of infection. Both assays performed equivalently and proved to be more sensitive than traditional culture methods, detecting O. ophiodiicola in 98% of the culture-positive samples and in 40% of the culture-negative snakes that had clinical signs of SFD. In addition, the assays did not cross-react with a panel of 28 fungal species that are closely related to O. ophiodiicola or that commonly occur on the skin of snakes. The assays did, however, indicate that some asymptomatic snakes (~6%) may harbor low levels of the fungus, and that PCR should be paired with histology when a definitive diagnosis is required. CONCLUSIONS: These assays represent the first published methods to detect O. ophiodiicola by real-time PCR. The ITS assay has great utility for assisting with SFD diagnoses whereas the IGS assay offers a valuable tool for research-based applications.

Invasive pulmonary infection caused by Chrysosporium articulatum: the first case report.

Chrysosporium species, saprobic soil fungi, comprise more than 60 species. There is some confusion regarding the taxonomy and nomenclature between Chrysosporium and Emmonsia since the causative agents of adiaspiromycosis, the development of big thick-walled spores (adiaspores) in humans or animals, were previously thought to be Chrysosporium. Chrysosporium articulatum has never been reported to cause invasive infection in humans. We report herein the first case of invasive pulmonary infection caused by Chrysosporium articulatum in a 16-year-old man with acute T-cell lymphoblastic leukaemia. He was successfully treated with voriconazole.