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Tabanomyces milkoi (Dudka and Koval) emended, genus novum, a fungal pathogen of horseflies.

A fungus originally described as a new species of Coelomomyces (C. milkoi) from larval Tabanidae in the Ukraine, USSR, is transferred to a new genus, Tabanomyces, and described in detail. The zygospores germinate to form a linear four-celled conidiophore, each cell producing a spherical conidium ejected as in Conidiobolus, thus confirming that the organism belongs in the Entomophthorales. The resemblance between the conidiophore of Tabanomyces milkoi and the basidium of the rusts supports the old concept that the rust fungi may be related to the Entomophthorales.

Journal Article↗

Pathology of equine phycomycosis.

The pathological findings in three cases of equine phycomycosis in Costa Rica are described. Two cutaneous (Pythium sp) and one nasal lesion (Conidiobolus) were observed.

Animals↗

Fluconazole in the therapy of tropical deep mycoses.

A clinical study was conducted to test the efficacy of fluconazole in the treatment of tropical deep mycoses. Two out of four patients with zygomycosis due to Conidiobolus coronatus who were treated with the drug were completely cured; the other two patients exhibited considerable improvement but could not be followed up. Two patients with eumycetoma, one due to an Acremonium sp. and one due to Pseudallescheria boydii, were treated successfully, whereas another patient with a eumycetoma caused by an unidentified fungus could not be followed up. A complete cure was achieved with one patient with African histoplasmosis and one with candiduria. A case of cerebral phaeohyphomycosis due to Cladosporium sp. showed some improvement but the patient later developed meningitis and died.

Adolescent↗

Mucormycosis and entomophthoramycosis: a review of the clinical manifestations, diagnosis and treatment.

The class Zygomycetes is divided into two orders, Mucorales and Entomophthorales. These two orders produce dramatically different infections. Genera from the order Mucorales (Rhizopus, Mucor, Rhizomucor, Absidia, Apophysomyces, Cunninghamella and Saksenaea) cause an angioinvasive infection called mucormycosis. Mucormycosis presents with rhino-orbito-cerebral, pulmonary, disseminated, cutaneous, or gastrointestinal involvement. Immunocompromising states such as haematological malignancy, bone marrow or peripheral blood stem cell transplantation, neutropenia, solid organ transplantation, diabetes mellitus with or without ketoacidosis, corticosteroids, and deferoxamine therapy for iron overload predispose patients to infection. Mucormycosis in immunocompetent hosts is rare, and is often related to trauma. Mortality rates can approach 100% depending on the patient's underlying disease and form of mucormycosis. Early diagnosis, along with treatment of the underlying medical condition, surgery, and an amphotericin B product are needed for a successful outcome. Genera from the order Entomophthorales produce a chronic subcutaneous infection called entomophthoramycosis in immunocompetent patients. This infection occurs in tropical and subtropical climates. The genus Basidiobolus typically produces a chronic subcutaneous infection of the thigh, buttock, and/or trunk. Rarely, it has been reported to involve the gastrointestinal tract. The genus Conidiobolus causes a chronic infection of the nasal submucosa and subcutaneous tissue of the nose and face. This paper will review the clinical manifestations, diagnosis and treatment of mucormycosis and entomophthoramycosis.

Humans↗

Clinical observations on equine phycomycosis.

During a clinical study of equine phycomycosis in tropical northern Australia 3 specific forms of phycomycosis were identified. Of 266 cases diagnosed in 5 different laboratories, hyphomycosis caused by Hyphomyces destruens was responsible for 76.7%, basidiobolomycosis caused by Basidiobolus haptosporus for 18.0%, and entomophthoramycosis caused by Conidiobolus coronatus for 5.3%. Most cases of hyphomycosis were observed between March and July, that is after the monsoonal wet summer, but were calculated from clinical histories to originate in the wet season between November and May. Basidiobolomycosis and entomophthoramycosis occurred regularly during the year and no seasonal incidence could be determined. All of these conditions occurred in horses in similar tropical regions of Queensland and the Northern Territory. No breed, sex or age predisposition was found but significant differences between the 3 fungal diseases in site of infection and appearance and size of the lesion were observed. Hyphomycosis was seen as an ulcerative granuloma containing characteristic coral-like kunkers. Lesions were most commonly found on the legs and ventral parts of the chest and abdomen, areas where contact with swamp water was apparent. Basidiobolomycosis was clinically similar to hyphomycosis but was mostly found on the lateral parts of the trunk and face where contact with contaminated soil might be expected. All lesions of entomophthoramycosis were located in the nasal region thus suggesting infection by inhalation.

Animals↗

Treatment of mycotic rhinitis with itraconazole in three horses.

Itraconazole, a third-generation azole, was evaluated for treatment of resistant nasal mycotic infections in horses. Two horses with Aspergillus spp nasal granulomas and 1 horse with Conidiobolus coronatus nasal infection were treated with itraconazole (3 mg/kg PO bid). One of the horses with nasal aspergillosis was also treated by surgical resection of the nasal septum. The treatment time for the horses ranged from 3 to 4.5 months. No adverse effects were noted in any of the horses during the treatment period. Peak and trough serum itraconazole concentrations were < 0.5 micrograms/mL in all 3 horses. Itraconazole (3 mg/kg PO bid) appears to be effective in the treatment of nasal Aspergillus spp infections in horses because the fungal infection was eliminated in both horses. One horse still had excessive nasal sounds during exercise and was retired from training, whereas the other horse returned to normal. The nasal C. coronatus infection appeared resistant to itraconazole treatment in the affected horse because the granulomas were still present after 4.5 months of treatment.

Animals↗

Zygomycetes in human disease.

The Zygomycetes represent relatively uncommon isolates in the clinical laboratory, reflecting either environmental contaminants or, less commonly, a clinical disease called zygomycosis. There are two orders of Zygomycetes containing organisms that cause human disease, the Mucorales and the Entomophthorales. The majority of human illness is caused by the Mucorales. While disease is most commonly linked to Rhizopus spp., other organisms are also associated with human infection, including Mucor, Rhizomucor, Absidia, Apophysomyces, Saksenaea, Cunninghamella, Cokeromyces, and Syncephalastrum spp. Although Mortierella spp. do cause disease in animals, there is no longer sufficient evidence to suggest that they are true human pathogens. The spores from these molds are transmitted by inhalation, via a variety of percutaneous routes, or by ingestion of spores. Human zygomycosis caused by the Mucorales generally occurs in immunocompromised hosts as opportunistic infections. Host risk factors include diabetes mellitus, neutropenia, sustained immunosuppressive therapy, chronic prednisone use, iron chelation therapy, broad-spectrum antibiotic use, severe malnutrition, and primary breakdown in the integrity of the cutaneous barrier such as trauma, surgical wounds, needle sticks, or burns. Zygomycosis occurs only rarely in immunocompetent hosts. The disease manifestations reflect the mode of transmission, with rhinocerebral and pulmonary diseases being the most common manifestations. Cutaneous, gastrointestinal, and allergic diseases are also seen. The Mucorales are associated with angioinvasive disease, often leading to thrombosis, infarction of involved tissues, and tissue destruction mediated by a number of fungal proteases, lipases, and mycotoxins. If the diagnosis is not made early, dissemination often occurs. Therapy, if it is to be effective, must be started early and requires combinations of antifungal drugs, surgical intervention, and reversal of the underlying risk factors. The Entomophthorales are closely related to the Mucorales on the basis of sexual growth by production of zygospores and by the production of coenocytic hyphae. Despite these similarities, the Entomophthorales and Mucorales have dramatically different gross morphologies, asexual reproductive characteristics, and disease manifestations. In comparison to the floccose aerial mycelium of the Mucorales, the Entomophthorales produce a compact, glabrous mycelium. The asexually produced spores of the Entomophthorales may be passively released or actively expelled into the environment. Human disease with these organisms occurs predominantly in tropical regions, with transmission occurring by implantation of spores via minor trauma such as insect bites or by inhalation of spores into the sinuses. Conidiobolus typically infects mucocutaneous sites to produce sinusitis disease, while Basidiobolus infections occur as subcutaneous mycosis of the trunk and extremities. The Entomophthorales are true pathogens, infecting primarily immunocompetent hosts. They generally do not invade blood vessels and rarely disseminate. Occasional cases of disseminated and angioinvasive disease have recently been described, primarily in immunocompromised patients, suggesting a possible emerging role for this organism as an opportunist.

Fungi↗

Immunodiffusion test for serodiagnosing subcutaneous zygomycosis.

Culture filtrate antigens of Basidiobolus ranarum and Conidiobolus coronatus were analyzed by immunodiffusion (ID) with homologous rabbit antisera. B. ranarum and C. coronatus were each found to have five specific antigens. Results of tests with heterologous antisera indicated that all of the species shared at least one antigen. ID tests incorporating the specific precipitin bands as references were developed for detection of basidiobolomycosis and conidiobolomycosis. These tests were performed with sera from humans and horses with proven basidiobolomycosis and conidiobolomycosis as well as with control sera from humans and animals with and without heterologous mycotic and oomycotic infections. Only sera from cases of basidiobolomycosis and conidiobolomycosis produced lines of identity with the reference precipitates of B. ranarum and C. coronatus, respectively. The ID tests were found to be completely sensitive and specific for determining the etiology of zygomycosis caused by these two species. In addition they appeared useful for monitoring resolution of the infections.

Animals↗

Immunoblot analysis of the humoral immune response to Pythium insidiosum in horses with pythiosis.

Reactions to Pythium insidiosum by sera from horses with active pythiosis were investigated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting. Five strains of P. insidiosum were grown in nutrient broth and then sonicated. After centrifugation, supernatant antigens were separated by SDS-PAGE. An exoantigen of Conidiobolus coronatus was also tested. Bands with molecular weights between 97,000 and 14,000 were identified by Coomassie blue and silver staining. After being transferred to nitrocellulose, the antigens were reacted against sera from six horses with pythiosis, sera from four horses cured a year earlier by vaccination, and sera from five healthy horses. The sera from horses with pythiosis recognized at least 20 antigens in all strains. Three antigens with molecular weights of 32,000, 30,000, and 28,000 appeared to be immunodominant and specific. Sera from horses cured by immunotherapy showed only five very weak bands, three of them the 32,000-molecular-weight (32K), 30K, and 28K antigens. No bands were observed with sera from healthy horses or sera from horses with a variety of other infections. Sera from horses with pythiosis cross-reacted with the 44K antigen of C. coronatus. The immunodominant antigens described here may be useful for diagnostic purposes and in immunotherapy for this oomycotic infection in horses.

Animals↗

Development of a species-specific probe for Pythium insidiosum and the diagnosis of pythiosis.

Pythium insidiosum, the only species in the genus that infects mammals, is the etiological agent of pythiosis, a granulomatous disease characterized by cutaneous and subcutaneous lesions and vascular diseases. Accurate diagnosis of pythiosis and identification of its causal agent are often inconsistent with current immunological diagnostic methods. A species-specific DNA probe was constructed by using a 530-bp HinfI fragment from the ribosomal DNA intergenic spacer of P. insidiosum. When the probe was incubated with dot blots of genomic DNA from 104 Pythium species, it hybridized only to the DNA of P. insidiosum and P. destruens-two species that have been considered conspecific. The probe also hybridized to DNA from 22 P. insidiosum isolates in this study, regardless of their geographic origin or animal host. When tested against genomic DNA from other pathogenic organisms (Aspergillus fumigatus, Basidiobolus ranarum, Conidiobolus coronatus, Lagenidium giganteum, Paracoccidioides brasiliensis, and Prototheca wickerhamii), no cross-hybridization of the probe was detected. The specificity of the probe to hybridize to genomic DNA from all isolates of P. insidiosum and not cross-react with DNA from other Pythium species or pathogens that cause symptoms similar to pythiosis in their hosts makes it a powerful tool for the accurate diagnosis of pythiosis. In addition, the probe has the potential for pathological and environmental diagnostic applications.

DNA Probes↗

Entomophthoramycosis: therapeutic success by using amphotericin B and terbinafine.

A 12-year-old girl had been presenting a woody infiltration and erythema in the frontal region and on the entire left half of the face, leading to deformity of the nose and buccal fissure, and adenomegaly in a posterior cervical chain, for the last 18 months. Sinusitis was diagnosed and treated with antibiotics, and submitted to ethmoid sinusotomy, with no improvement. Several laboratory tests were made to find the correct diagnosis. An intradermal test for delayed hypersensitivity showed a positive reaction (5 mm) with necrosis for metabolic antigens for Conidiobolus. An oral mucosa biopsy showed a dense lymphohistiocytic infiltrate and focal points of necrosis. Gomori staining for fungi revealed countless wide, nonseptate hyphae. Amphotericin B was prescribed during 35 days, with no improvement. Terbinafine given orally was started in association with amphotericin B. Reduction of the lesions was observed 2 months later. No side effects were seen during 4 months of treatment.

Amphotericin B↗

The comparative pathology of equine cutaneous phycomycosis.

Pythiosis (204 cases, 77%), basidiobolomycosis (48 cases, 18%), and conidiobolomycosis (14 cases, 5%) were diagnosed morphologically from 266 horses with phycomycosis. All lesions were cutaneous ulcerative granulomas and three horses with pythiosis had metastatic lesions in regional lymph nodes. Lesions of pythiosis contained characteristic yellow, coral-like coagula and had a fibrotic surface containing sinus tract openings. Basidiobolomycosis was characterized by infrequent small yellow coagula and a yellow line of fungal invasion beneath an edematous surface. Lesions of conidiobolomycosis had numerous small coagula and a nasal location. There were minor differences in inflammatory cell populations within the granulomatous lesions. Most differences were associated with coagula size and fungal morphology. Coagula were composed of collagen, fibrin, cellular debris, degranulated and whole eosinophils and hyphae. Histochemistry revealed no major differences among the three diseases. Pythium sp hyphae were 2.6 to 6.4 micrometers in diameter, had thick walls, and occasionally were septate. Basidiobolus haptosporus hyphae were 5.1 to 20.5 micrometers in diameter, had thin walls, and commonly were septate. Conidiobolus coronatus hyphae were 5.1 to 12.8 micrometers in diameter, had thin walls, and commonly were septate. A perihyphal eosinophilic cuff ( Splendore - Hoeppli phenomenon) with a radius of up to 20 micrometers was associated with the latter two fungi. Ultrastructurally, Pythium sp was composed of a thick, single density cell wall while B. haptosporus and C. coronatus had thin, double-layered cell walls.

Animals↗

Development of a nested polymerase chain reaction assay for the detection and identification of Pythium insidiosum.

Pythium insidiosum is an important cause of cutaneous and gastrointestinal disease in horses and dogs in the southeastern United States. Culture-based diagnosis of pythiosis is rarely definitive because production and identification of reproductive structures is difficult. The purpose of this study was to develop a polymerase chain reaction (PCR)-based assay for the identification of P insidiosum. Genomic DNA was extracted from 3 clinical isolates of P insidiosum and I isolate each of Pythium graminicola and Pythium arrhenomanes. The ITS I region of the ribosomal RNA gene of each isolate was amplified and sequenced, and the resultant sequences were aligned with published sequences for Pythium aphanidermatum, P acanthicum, and P myriotylum. A pair of P insidiosum-specific primers (PI-1 and PI-2) were designed from variable regions within the ITSI region. A nested PCR assay was developed in which the 1st round amplified the ITSI region by use of universal fungal primers. Second-round amplification utilized the internal P insidiosum-specific primers PI-1 and PI-2. Specificity of the assay was tested with DNA extracted from cultures of the following: 10 clinical isolates of P insidiosum and 1 isolate each of P graminicola, P irregulare, P arrhenomanes, P myriotylum, P deliense, Basidiobolus ranarum, Conidiobolus coronatus, Aspergillus terreus, Lagenidium giganteum, and a canine-pathogenic Lagenidium species. Nested PCR produced a single 105-base pair amplicon for each of the P insidiosum isolates, but did not produce amplicons for any of the other isolates. Results of this study suggest that PCR is a useful tool for the identification of P insidiosum.

Animals↗

Rhinoentomophthoromycosis: report of the first two cases observed in Costa Rica (Central America), and review of the literature.

The first two cases of rhinoentomophthoromycosis to be recognized in Costa Rica are reported. The first patient was a 32-year-old Caucasian male from the Pacific Coast, and the second, a 17-year-old Negro male from the Atlantic Coast. Both cases showed the typical involvement of the nasofacial skin with the formation of subcutaneous nodules. One patient also showed left maxillary sinus involvement. Both patients were in general good health, without any associated disease. Cultures from the second patient, taken from the glabellar nodule, were positive for Conidiobolus coronatus. These two cases represent the first documentation of this uncommon mycosis in Central America.

Adolescent↗

Entomophthoromycosis in Maharaj Nakorn Chiang Mai Hospital.

OBJECTIVE: To review documented entomophthoromycosis patients at Maharaj Nakorn Chiang Mai Hospital from 1985 to 2001. MATERIAL AND METHOD: A retrospective review was performed at Maharaj Nakorn Chiang Mai Hospital from 1985 to 2001. Eight cases of entomophthoromycosis were found between 1988 and 1993, with five patients diagnosed as subcutaneous zygomycosis, I GI entomophthoromycosis and 2 rhinofacial zygomycosis. RESULTS: The subcutaneous zygomycosis group was composed of 5 female patients, aged 7- 77 years. They presented with a painless subcutaneous mass, which was solitary or multiple and most commonly found on the extremities. The duration of disease was between 3 months to 5 years. The cultures grew Basidiobolus ranarum. A case of GI entomophthoromycosis was seen in a 34 year old man, who presented with dyspepsia, multiple intraabodminal masses and peritonitis. Two cases of conidiobolomycosis (rhinofacial zygomycosis) were seen. These two patients were male, 26 and 39 years old, and they presented with unilateral nasal obstruction from a mass at the inferior turbinate. The cultures grew Conidiobolus coronatus. CONCLUSION: Entomophthoromycosis in the northern part of Thailand is rare. The disease should be differentially diagnosed by a chronic painless tumor. The patients in this review responded very well to the recommended therapy.

Adolescent↗

Entomophthoramycosis conidiobolae in a llama.

A 9-year-old female llama was evaluated for chronic dermatosis involving the external nares. The condition had been diagnosed as a fungal infection. Examination of punch biopsy specimens confirmed the diagnosis of Conidiobolus coronatus. Because of the chronicity of the disease and history of ineffective drug treatment, excision of infected tissue was attempted. Permanent nasal openings were created by suturing the nasal mucosa to the skin after debulking of infected tissue. Excision of most of the infected tissue enabled the llama to breathe easily for 3 years. Because C coronatus does not usually induce deep infections, it may continue to remain dormant in this llama.

Animals↗

[5 new cases of entomophthoromycosis observed in Cameroon. Role of immunofluorescence in thr diagnosis].

The authors present five new cases of entomophthoromycosis observed during three years of histopathology in Cameroon. Three classic cases of rhinoentomophthoromycosis. In one case, Conidiobolus coronatus was isolated. Two cases of subcutaneous entomophthoromycosis, without isolation of the germ. In one observation, the clinical and histological aspect suggested mucormycosis post entomophthoromycosis. An immunofluorescence technique showed the Basidiobolus haptosporus to be the real cause.

Adult↗

[Histology of insect infection by entomopathogenic fungi: interest in a new procedure for the study of precocious phases].

Infective and invasive processes of entomopathogenic fungi expected to be used in biological control shall be analysed in detail. Histological and ultrastructural studies are therefore essential. This paper is devoted to the field of light microscopy; its purpose is to show that coating, in a 4% agar solution, the insects infected by a fungus, prior to fixation and embedding, allows the preservation of the fungal structures present on the cuticle, especially the germinating conidia. This is useful when observing the initial stages of infection. The relevance of this coating technique is illustrated with the example of Pea aphid, Acyrthosiphon pisum, infected by Conidiobolus osmodes. This contribution is the first histological study of infective processes of this entomophthoralean fungus in an insect.

Animals↗