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ABSTRACTIn recent years, the Chrysosporiumanamorph of Nannizziopsis vriesii(CANV), Chrysosporium guarroi, Chrysosporium ophiodiicola, and Chrysosporiumspecies have been reported as the causes of dermal or deep lesions in reptiles. These infections are contagious and often fatal and affect both captive and wild animals. Forty-nine CANV isolates from reptiles and six isolates from human sources were compared with N. vriesiibased on their cultural characteristics and DNA sequence data. Analyses of the sequences of the internal transcribed spacer and small subunit of the nuclear ribosomal gene revealed that the reptile pathogens and human isolates belong in well-supported clades corresponding to three lineages that are distinct from all other taxa within the family Onygenaceaeof the order Onygenales. One lineage represents the genus Nannizziopsisand comprises N. vriesii, N. guarroi, and six additional species encompassing isolates from chameleons and geckos, crocodiles, agamid and iguanid lizards, and humans. Two other lineages comprise the genus Ophidiomyces, with the species Ophidiomyces ophiodiicolaoccurring only in snakes, and Paranannizziopsisgen. nov., with three new species infecting squamates and tuataras. The newly described species are Nannizziopsis dermatitidis, Nannizziopsis crocodili, Nannizziopsis barbata, Nannizziopsis infrequens, Nannizziopsis hominis, Nannizziopsis obscura, Paranannizziopsis australasiensis, Paranannizziopsis californiensis, and Paranannizziopsis crustacea. Chrysosporium longisporumhas been reclassified as Paranannizziopsis longispora. N. guarroicauses yellow fungus disease, a common infection in bearded dragons and green iguanas, and O. ophiodiicolais an emerging pathogen of captive and wild snakes. Human-associated species were not recovered from reptiles, and reptile-associated species were recovered only from reptiles, thereby mitigating concerns related to zoonosis.

ABSTRACTWe report the first case of Neosartorya laciniosainvasive sinusitis involving the orbit in an immunocompromised male with aplastic anemia. Treatment included surgical debridement with enucleation of the eye and combination voriconazole and micafungin therapy followed by voriconazole alone. The fungus was identified using sequencing of partial benAand calmodulin genes.

ABSTRACTAn outbreak of disseminated granulomatous disease occurred in a group of veiled chameleons (Chamaeleo calyptratus) in a zoo collection. An adult female and six offspring developed large granulomas in multiple organs and were euthanized. At necropsy, roughly spherical yellow-to-white nodules 1 to 3 mm in diameter were grossly visible in the liver and other organs. Histopathology revealed fungal elements that were spherical to ovoid in shape, fragments of slender to irregularly swollen hyphae, and occasional conidia produced on phialides. Fungal isolates were initially suspected on the basis of morphology results to represent Paecilomyces viridis, a species known only from one outbreak of fatal mycosis in carpet chameleons (Furcifer lateralis). Data obtained from morphological studies and from phylogenetic analyses of nuclear ribosomal rRNA (rDNA) sequence data revealed the Danish chameleon isolates to be a related undescribed anamorphic species within the family Clavicipitaceae that includes many insect pathogens. Chamaeleomyces granulomatisgen. et sp. nov. is given as the name for the newly described fungus, and P. viridisis transferred to the new genus as Chamaeleomyces viridiscomb. nov. Chamaeleomycesspecies are distinguished by having basally swollen phialides tapering to a narrow neck, conidia in fragile chains, and pale green to greenish-gray colonies. Both species are dimorphic, producing a transitory yeast stage characterized by ovoid-to-subglobose or subcylindrical yeast-like cells. Chamaeleomycesspecies appear to be rare but aggressive pathogens of chameleons.

Yellow rot, caused by an ascomycetous fungus having a distinctive arthroconidial anamorph, is the most destructive disease of cultivated Ganoderma lucidumin Korea, but the identity of the yellow rot pathogen (YRP) remains uncertain. Isolates have been identified as Xylogone sphaerospora(with putative anamorph Sporendonema purpurascens) or as Arthrographis cuboidea. Therefore we used morphological features, pathogenicity tests and phylogenetic analyses of DNA sequences from the nuclear ribosomal genes, including partial small subunit and internal transcribed spacer regions, and from the gene encoding RNA polymerase second largest subunit to evaluate the relationship between YRP isolates and these species. YRP isolates formed a distinct subgroup within a clade that included X. sphaerospora, A. cuboideaand Scytalidium lignicola, the type species of Scytalidium, but the disposition of the clade within the Leotiomycetes was uncertain. We describe Xylogone ganodermophthorasp. nov. and Scytalidium ganodermophthorumsp. nov. for the teleomorph and anamorph of YRP respectively. Arthrographis cuboideais reclassified as Scytalidium cuboideumcomb. nov., and the anamorph of X. sphaerosporais named Scytalidium sphaerosporumsp. nov. In pathogenicity tests only X. ganodermophthoracaused disease in Ganoderma lucidum. Amplified fragment length polymorphism analyses showed that X. ganodermophthorapopulations from diseased fruiting bodies or from oak wood in Korea consisted of two clonal groups.

ABSTRACTWe report the first case of infection caused by Graphium basitruncatumin a man with acute leukemia who developed persistent fungemia and skin lesions. G. basitruncatum, a member of the Microascaceae, is phylogenetically and morphologically distinct from Graphium penicillioidesand the opportunistic pathogens Scedosporium apiospermum(Pseudallescheria boydii) and Scedosporium prolificans.

We report the first case of infection caused by Graphium basitruncatum in a man with acute leukemia who developed persistent fungemia and skin lesions. G. basitruncatum, a member of the Microascaceae, is phylogenetically and morphologically distinct from Graphium penicillioides and the opportunistic pathogens Scedosporium apiospermum (Pseudallescheria boydii) and Scedosporium prolificans.

ABSTRACTA case of primary cutaneous zygomycosis caused by Mucor circinelloidesis described. Histopathology showed typical hyphae along with chlamydospores. The isolate was identified by molecular and phenotypic methods. The utility of sequence analysis of the internal transcribed spacer region is highlighted; however, further studies are needed to assess species genetic heterogeneity.

A case of primary cutaneous zygomycosis caused by Mucor circinelloides is described. Histopathology showed typical hyphae along with chlamydospores. The isolate was identified by molecular and phenotypic methods. The utility of sequence analysis of the internal transcribed spacer region is highlighted; however, further studies are needed to assess species genetic heterogeneity.

ABSTRACTFungal peritonitis due to Curvulariaspecies in patients undergoing peritoneal dialysis is a very rare problem. We report a case of peritonitis caused by Curvularia inaequalis. This is the first report in the English literature of this species causing human infection. We also review the six previously reported cases of continuous ambulatory peritoneal dialysis peritonitis caused by other Curvulariaspecies.

Sterile fungi isolated from surface-sterilized roots of the Ericaceae,and hypothesized to be conspecific based primarily on restriction fragment length polymorphisms, were provisionally named as Variable White Taxon (VWT). In preliminary resynthesis trials with Vaccinium myrtilloidesor V. vitis-idaea,isolates did not form typical ericoid mycorrhizas. Additional isolates obtained from roots of the Orchidaceae, Pinaceae, Betulaceaeand Salicaceae,and given informal names such as Sterile white 1 (SW1), were thought to represent the same taxon based on cultural similarities. To evaluate conspecificity and infer phylogenetic affinities, partial nuclear ribosomal DNA sequences were determined. Parsimony analyses supported a species level distinction for VWT/SW1 and indicated that the fungus is placed within the species complex referred to as the “Hymenoscyphus ericaeaggregate” which includes H. ericae(Leotiomycetes), many unnamed taxa and Cadophora finlandica.The new genus and species Meliniomyces variabilisis proposed to accommodate this root-associated fungus to facilitate discussion and information retrieval, and to provide a foundation for additional experimental work. Although isolates are sterile in culture, they can be identified by morphological characters in conjunction with ribosomal internal transcribed spacer (ITS) sequence data. The mycorrhizal status of M. variabilisis not yet clear. In prior published results, strains demonstrated no or some colonization with ericoid and ectomycorrhizal hosts but did not form true ectomycorrhizas. ITS analyses indicated that the “H. ericaeaggregate” includes several other well-supported clades putatively named as Meliniomycesspecies. Representative strains were examined morphologically for two of these species, described as M. vraolstadiaeand M. bicolor.Both include ectomycorrhizal mycobionts of the “Piceirhiza bicolorata” morphotype. Rhizoscyphus ericaeis accepted as the appropriate name for H. ericae.

Cryptosporiopsisspecies are anamorphs of ascomycetes in the genera Peziculaand Neofabraea(Dermataceae). These fungi are occasionally isolated from roots of woody plants but may be difficult to identify due to absence of sporulation. Some isolates obtained from roots of ericaceous hosts had previously been linked phylogenetically to Peziculaand, when regrown, revealed conidiomata and conidia typical of Cryptosporiopsisspecies. Cultural and molecular data allowed for the recognition of the new species, Cryptosporiopsis ericaeand C. brunnea.

A simple cultural method was investigated for its reliability in distinguishing the ericoid mycobiont Oidiodendronmaius from selected other species of Oidiodendron.Forty three isolates were grouped by morphology after 28 d growth on cereal agar overlaid with a cellophane membrane. All isolates of O. maiusand its close relative O. citrinumexpressed characteristic colonial morphologies allowing recognition regardless of sporulation. Isolates grouped by colonial features correlated with strongly supported groupings obtained by analysis of nuclear ribosomal internal transcribed spacer (ITS) region sequences, including O. maiuswith O. citrinum, O. griseumwith O. flavum,and O. truncatumas an independent group. Isolates of O. tenuissimum,including the ex-type of the purported synonym O. fuscum,demonstrated cultural variation and were dispersed among several different groups in the ITS analysis. O. fuscumis here regarded as a distinct taxon.

Phylogenies inferred from the analysis of DNA sequence data have shown that the Onygenales contains clades that do not correspond with previously described families. One lineage identified in recent molecular phylogenetic studies includes the dimorphic pathogens belonging to the genera Ajellomyces, Emmonsiaand Paracoccidioides.To evaluate the degree of support for this lineage and determine whether it includes additional taxa, we examined relationships among the members of this clade and selected saprobic onygenalean taxa based on maximum-parsimony analyses of partial nuclear large RNA subunit (LSU) and internal transcribed spacer (ITS) sequences. A clade distinct from the Onygenaceae was found to encompass Ajellomyces(including the anamorph genera Blastomyces, Emmonsiaand Histoplasma) and Paracoccidioides brasiliensis.The members of this lineage are saprobic and pathogenic vertebrate-associated taxa distinguished by their globose ascomata with coiled appendages, muricate globose or oblate ascospores, and lack of keratinolytic activity. Anamorphs are solitary aleurioconidia or irregular alternate arthroconidia. Based on molecular data and on morphological and physiological similarities among these taxa, we propose the new family, Ajellomycetaceae.

ABSTRACTWe describe allergic fungal sinusitis caused by Trichoderma longibrachiatumin a patient with a history of atopy and asthma. A Gram stain of a sinus biopsy specimen was initially thought to contain yeast cells, but when Trichodermawas recovered in culture, these cells were subsequently recognized as chlamydospores. The patient was successfully managed with a combination of sinus lavage, oral corticosteroids, itraconazole, and allergen immunotherapy. This case also points out that careful scrutiny of direct smears is required to ensure that fungal structures are not misinterpreted.

ABSTRACTThe Chrysosporiumanamorph of Nannizziopsis vriesii(CANV) is a fungus that has been implicated in several recent cases of reptile dermatomycoses. A survey was conducted to investigate whether this fungus was present on the skins of healthy squamate reptiles. Skin was collected as aseptically as possible from actively shedding lizards (n 36) or from freshly shed snake exuvia (n 91) and placed on fungal culture media for selective recovery of cycloheximide-tolerant fungi. The CANV was cultured from only one animal, an African rock python, Python sebae. Fungi belonging to 50 genera were identified from 127 reptiles: Aspergillusspp., Penicilliumspp., and Paecilomyces lilacinuswere most frequently isolated. Keratinophilic fungi isolated from reptiles did not belong to zoophilic or anthropophilic species, inferring that the potential for acquisition of dermatophytosis from handling squamate reptiles is low.

Five fungal isolates, tentatively identified as belonging to Thysanophora, were re-examined to confirm their generic placement. DNA sequences from multiple loci were determined and compared to homologous sequences from other fungi. The Thysanophora-like isolates belong in the genus Penicillium on the basis of phylogenetic analysis of their gene sequences. Because these isolates have unique phylogenetic positions within Penicillium and have unique phenotypes we describe them as the new species P. boreae, P. subarcticum and P. canariense spp. nov. In addition, P. pullum sp. nov. is described for NRRL 721, which served as the typical isolate of P. fuscum in the monographic studies of Raper & Thom and Ramirez; however, the name P. fuscum has been applied and neotypified using a different specimen and NRRL 721 is markedly different phylogenetically from the neotype of P. fuscum.

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

AbstractScopulariopsisanamorphs are known for many species of the genus Microascus(Ascomycota, Microascaceae), but teleomorph connections for anamorphic species within the ‘Scopulariopsis brevicaulisseries' are tenuous or lacking. Microascus brevicauliswas recently described as the teleomorph of the type and commonest species, S. brevicaulis, but only a few isolates yielded fertile perithecia. To investigate whether paucity of sexual reproduction was the result of heterothallism, mating experiments were conducted among isolates representing the species S. brevicaulis, S. Candida, S. asperula, S. fuscaand S. koningii.Results demonstrated heterothallism within three species and confirmed that two taxa could be reduced to synonymy. Three holomorph species, M. brevicaulis, M. manginii, and M. niger, are recognized to include anamorphs S. brevicaulis(synonym S. koningii), S. Candida, and S. asperula(synonyms S. arnoldii, S. bestae, S. fusca, and S. roseola), respectively. Microascus nigeris redescribed and a neotype proposed. The three species are most readily recognized by colony color (sandy tan to avellaneous in M. brevicaulis, white to cream in M. manginii, and medium to dark fuscous brown in M. niger). These colonial distinctions correlate generally with conidia that are coarsely roughened, smooth, or finely roughened, respectively. However, conidium ornamentation, previously considered a reliable taxonomic character, is shown to be variable.

Abstract: Decayed wood from fallen white spruce (Picea glauca) and trembling aspen (Populus tremuloides) collected in northeastern Alberta, Canada, was the source of new isolates of species in the ascomycete generaGymnostellatospora andPseudogymnoascus. In addition to new reports ofG. japonica, G. frigida andP. roseus, two new species are described.Gymnostellatospora canadensis sp. nov. resemblesG. japonica but differs in producing brown ascomata and in the formation of an arthroconidial anamorph.Gymnostellatospora subnuda sp. nov. is distinct in lacking differentiated peridial hyphae.Gymnostellatospora alpina was not found in decayed wood but is reviewed based on extralimital material. A dichtomous key to the five species ofGymnostellatospora is provided.

Decayed wood from fallen white spruce (Picea glauca)and trembling aspen (Populus tremuloides)collected in northeastern Alberta, Canada, was the source of new isolates of species in the ascomycete genera Gymnostellatosporaand Pseudogymnoascus.In addition to new reports of G. japonica, G. frigidaand P. roseus, two new species are described. Gymnostellatospora canadensissp. nov. resembles G.japonicabut differs in producing brown ascomata and in the formation of an arthroconidial anamorph. Gymnostellatospora subnudasp. nov. is distinct in lacking differentiated peridial hyphae. Gymnostellatospora alpinawas not found in decayed wood but is reviewed based on extralimital material. A dichotomous key to the five species of Gymnostellatosporais provided.

ABSTRACTWe report the first case of invasive pulmonary infection caused by the thermotolerant ascomycetous fungus Gymnascella hyalinosporain a 43-year-old female from the rural midwestern United States. The patient was diagnosed with acute myelogenous leukemia and treated with induction chemotherapy. She was discharged in stable condition with an absolute neutrophil count of 100 cells per µl. Four days after discharge, she presented to the Cancer Clinic with fever and pancytopenia. A solitary pulmonary nodule was found in the right middle lobe which was resected by video-assisted thoracoscopy (VATHS). Histopathological examination revealed septate branching hyphae, suggesting a diagnosis of invasive aspergillosis; however, occasional yeast-like cells were also present. The culture grew a mold that appeared dull white with a slight brownish tint that failed to sporulate on standard media. The mold was found to be positive by the AccuProbe Blastomyces dermatitidisCulture ID Test (Gen-Probe Inc., San Diego, Calif.), but this result appeared to be incompatible with the morphology of the structures in tissue. The patient was removed from consideration for stem cell transplant and was treated for 6 weeks with amphotericin B (AmB), followed by itraconazole (Itr). A VATHS with biopsy performed 6 months later showed no evidence of mold infection. In vitro, the isolate appeared to be susceptible to AmB and resistant to fluconazole and 5-fluorocytosine. Results for Itr could not be obtained for the case isolate due to its failure to grow in polyethylene glycol used to solubilize the drug; however, MICs for a second isolate appeared to be elevated. The case isolate was subsequently identified as G. hyalinosporabased on its formation of oblate, smooth-walled ascospores within yellow or yellow-green tufts of aerial hyphae on sporulation media. Repeat testing with the Blastomycesprobe demonstrated false-positive results with the case isolate and a reference isolate ofG. hyalinospora. This case demonstrates that both histopathologic and cultural features should be considered for the proper interpretation of this molecular test and extends the list of fungi recognized as a cause of human mycosis in immunocompromised patients.

ABSTRACTWe present a case of maxillary sinusitis in a diabetic female caused by the basidiomycete fungus Schizophyllum commune. Identification of the isolate was hampered by its atypical features. Subcultures formed sterile medusoid structures from nonclamped mycelia until spontaneous dikaryotization resulted in the development of characteristic fan-shaped fruiting bodies. Identification was confirmed by the presence of spicules formed on the hyphae and by vegetative compatibility with known isolates.

Airborne fungi in honeybee overwintering and equipment cleaning facilities were enumerated and identified to determine worker exposure during cleaning and routine beekeeping operations. Testing was prompted by observations of extensive mold growth on dead bees and associated material and by results of a preliminary study at one Alberta beekeeping facility that showed very high numbers of mold colonies on air samples taken during worker activity. To evaluate whether high mold counts were indicative of a problem at a single site or were industry wide, approximately 120 air samples were collected with a Reuter centrifugal sampler inside 10 overwintering facilities before and during routine beekeeping activity during fall, winter, and spring periods. A set of 30 samples was collected from 15 sites used for annual equipment cleaning. This study showed that average spore counts per overwintering site ranged from 238 to 1442 colony-forming units (CFU)/m3 prior to disturbance by workers and from 2200 to 13,931 CFU/m3 while workers swept up dead bees. Levels of airborne molds recovered during annual cleaning of beekeeping equipment ranged from 300 to 54,700 CFU/m3 with an average of 16,083 CFU/m3. Potentially toxigenic, pathogenic, or allergenic molds were recovered at all sites. Since the data indicate that exposure to high levels of airborne molds is widespread throughout the industry, actions that might help minimize worker exposure are discussed.

In 1969, specialists at the Kananaskis hyphomycete workshop coined the terms ’blastic’ and ’thallic’ to describe two distinct modes of conidiogenesis. Since then, the original concepts have been slightly modified and redefined, and the terms have been widely adopted in taxonomic descriptions of conidial fungi. Problems arise in the application of these terms to conidial development in fungi which demonstrate morphological plasticity ranging from fragmentation of a hypha to extrusion of a portion of a hypha or cell. A number of fungi, such as anamorphs of Onygenales which includes many of the fungi pathogenic to man, demonstrate intergradations between blastic and thallic development. Because development in this group of fungi is difficult to categorize, it has led to an inconsistent treatment of taxa which share many other developmental features in common. In using these terms, it should be remembered that they represent extremes in a developmental spectrum.

Fatal pulmonary infection in a captive alligator (Alligator mississippiensis) is reported. At necropsy, the animal appeared to be in excellent nutritional condition, but a severe necrotizing bronchitis with bronchiectasis was present. Histological examination revealed numerous branched, septate, hyaline hyphae within the necrotic debris lining the bronchi and rarely infiltrating into the adjacent stroma. The fungus cultured from the lung was identified as F. moniliforme.

AbstractFive isolates of Scopulariopsis brevicaulishave been found to form perithecia in addition to typical conidia. The teleomorph is described as Microascus brevicaulissp. nov., characterized by small (70–130 μm diam), black, ostiolate ascocarps that are papillate or very short-necked, and with a peridium of textura angularis.The ascospores are reniform, 5–6 × 3.5-4.5 μm, smooth, and pale orange in mass. Although other species of Microascusproduce a Scopulariopsisstate in culture, this discovery now definitively links S. brevicaulis, the type species of Scopulariopsis, to the Microascaceae.

The metabolites of the wood rot decay fungus Sistotremaraduloides(P. Karst) Donk have been investigated and a new type of norpentaketide has been discovered. The compounds sistodiolynne (1), sistolynone (5), and sistopyrone (6) represent new carbon skeletons among natural products. The compounds are very unstable and readily polymerize in the presence of air to give black insoluble material. It has been shown by 13C NMR labelling experiments that they arise from five acetate units, and that a methyl carbon of one of the acetate units is lost during the biosynthesis. The structures were determined by spectroscopy methods, mainly NMR, and the absolute configuration of sistodiolynne was established by circular dichroism methods. Keywords: fungal metabolites, Sistotremaraduloides, norpentaketides, biosynthetic studies, polyacetylenes.

Thirtynine yeast isolates or dimorphic fungi were obtained from the hindgut of the lower termites Mastotermis darwiniensis(Mastotermitidae), Zootermopsis angusticollis, Z. nevadensis(Hodotermitidae), Neotermes jouteli(Kalotermitidae), Reticulitermes santonensis, Heterotermes indicola(Rhinotermitidae) and the roach Cryptocercus punctulatus. Using RAPD-PCR the 39 yeast isolates were assigned to 13 different species. Commonly yeast species were specific to the termite species isolates from. There were only two yeast species which were found in different species of lower termites. Based on phenotypic characters Debaryomyces hanseniishowed a high score in four species. The qualitative and quantitative yeast cell wall monosaccharide composition, the ubiquinone system, partial sequencing of 18S ribosomal DNA (bases 1273 to 948; numbering according to the gene of Saccharomyces cerevisiae), and the ultrastructure of septal pores indicate that 11 yeast species belong to the Endomycetales. Although ascospores were lacking, two of these species were identified to belong to the genus Debaryomyces. One remaining yeast isolate was identified as a Sporothrixanamorph representative for the filamentous Ascomycetes (Ophiostomataceae s. str.); the second species showed affinities to the Basidiomycetes in particular to the genus Trichosporon. Comparing an additional 18S rDNA fragment (bases 595 to 993) and RAPD-PCR data using different species type strains of the genus Sporothrix, the filamentous ascomycete was genotypically identified as Sporothrix albicans. Sporothrix. albicans, although phylogenetically closely related to S. schenckiivar. schenckiiand Ophiostoma stenocerasremains genotypically distinct. An emended species description of S. albicans is presented. Evidence is provided that the yeasts isolated from the hindgut can be considered symbionts.

The influence of microorganisms associated with alfalfa leafcutter bee (Megachile rotundata) larvae on chalkbrood caused by Ascosphaera aggregata was determined by comparing disease in agnotobiotic and axenic larvae, with and without an introduced microflora (gnotobiotic). Bacterial and fungal populations in the frass of larvae reared on natural provisions (agnotobiotic) were log 4.24 and log 3.90 cfu/g of frass, respectively. No fungi were recovered from the frass of larvae reared on γ-irradiated provisions, but bacterial populations averaged log 0.89 cfu/g (near-axenic). Following ingestion of ascospores of A. aggregata, the incidence of chalkbrood was significantly higher in agnotobiotic (67.6%) than near-axenic (50.2%) larvae, but the time to death of infected larvae was the same for both. Microorganisms were inoculated onto the surface of γ-irradiated provisions adjacent to leafcutter bee eggs and were ingested by larvae; 11 of 13 bacteria, 9 of 10 filamentous fungi, but 0 of the 7 yeasts were recovered from frass (gnotobiotic). Populations of inoculated bacteria and filamentous fungi recovered in frass ranged from log 5.40 to 7.30 cfu/g and log 5.21 to 6.52 cfu/g of frass, respectively. Of the 20 microorganisms recovered in frass, only Bacillus brevis and a Streptomyces sp. significantly increased the incidence of chalkbrood (74.0 and 74.8%, respectively) when larvae were coinoculated with ascospores of A. aggregata (1:1 ratio); the incidence of chalkbrood in larvae inoculated with the pathogen alone was 52.1%. None of the microorganisms themselves appeared to be deleterious to larvae. There were no significant differences in microbial populations in frass from healthy larvae or chalkbrood cadavers; however, noninoculated bacteria were associated with all nine fungal treatments. The results of this study suggest that microorganisms associated with the alimentary tracts of alfalfa leafcutter bee larvae have a minimal impact if any on chalkbrood. Copyright 1993, 1999 Academic Press

Characterization of microorganisms associated with alfalfa leaf-cutter bee (Megachile rotundata) nectar, pollen, provisions, larval guts, and frass (excreta) in Alberta demonstrated a varied aerobic microflora. Yeasts were isolated frequently from nectar, pollen, and provisions but rarely from guts or frass. The most prevalent yeast taxa were: Candida bombicola, Cryptococcus albidus, Metschnikowia reukaufii, and Rhodotorula glutinis. Although few filamentous fungi were found in nectar, they were frequently isolated from pollen and provisions; the predominant taxa were Alternaria alternata, Cladosporium cladosporioides, C. herbarum, Epicoccum nigrum, and Penicillium chrysogenum. Bacteria, including species of Bacillus, Corynebacterium, Micrococcus, and the actinomycete Streptomyces, also were prevalent in provisions and/or on pollen. In general, the diversity of microorganisms isolated from alimentary canals and frass was lower than from nectar, pollen, and provisions. Bacillus firmus, B. licheniformis, B. megaterium, B. pumilus, and Streptomyces spp. were the most frequently isolated bacteria, whereas Trichosporonoides megachiliensis was the most common filamentous fungus isolated from larval guts and/or frass. These taxa may be part of the resident microflora of the alimentary canal. Populations of bacteria and filamentous fungi, but not yeasts, were larger from Ascosphaera aggregata-infected larvae than from healthy larvae. However, with the exception of Aspergillus niger and T. megachiliensis in frass from healthy larvae, no taxon of filamentous fungi was conspicuously present or absent in infected larvae, healthy larvae, or their frass.

ABSTRACTWe report the first case of Chrysosporium zonatuminfection in a 15-year-old male with chronic granulomatous disease who developed a lobar pneumonia and tibia osteomyelitis while on prophylaxis with gamma interferon. The fungus was isolated from sputum and affected bone, and hyphae were observed in the bone by histopathology. Therapy with amphotericin B eradicated the osteomyelitis and pneumonia, but pneumonia recurred in association with pericarditis and pleuritis during therapy with itraconazole. These manifestations subsided, and no recurrences occurred with liposomal amphotericin B therapy. Infections caused by Chrysosporiumspecies are very rare, andC. zonatumhas not previously been reported to cause mycosis in humans. This species, the anamorph of the heterothallic ascomycete Uncinocarpus orissi(family Onygenaceae), is distinguished by its thermotolerance, by colonies which darken from yellowish white to buff, and by club-shaped terminal aleurioconidia borne at the ends of short, typically curved stalks. The case isolate produced fertile ascomata in mating tests with representative isolates. The median (range) MICs for our isolate as well as those for two other human isolates and a nonhuman isolate determined by the National Committee for Clinical Laboratory Standards method adapted for moulds were =0.06 µg/ml (=0.06 to 0.25 µg/ml) for amphotericin B, 0.687 µg/ml (0.25 to 2 µg/ml) for itraconazole, >128 µg/ml (>128 µg/ml) for flucytosine, and 48 µg/ml (32 to >128 µg/ml) for fluconazole.

ABSTRACTEmmonsia crescens, an agent of adiaspiromycosis,Blastomyces dermatitidis, the agent of blastomycosis, andHistoplasma capsulatum, the agent of histoplasmosis, are known to form meiotic (sexual) stages in the ascomycete genusAjellomyces(Onygenaceae,Onygenales), but no sexual stage is known for E. parva, the type species of the genus Emmonsia. To evaluate relationships among members of the putativeAjellomycesclade, large-subunit ribosomal and internal transcribed spacer region DNA sequences were determined from PCR-amplified DNA fragments. Sequences were analyzed phylogenetically to evaluate the genetic variation within the genus Emmonsiaand evolutionary relationships to other taxa. E. crescensand E. parvaare distinct species.E. crescensisolates are placed into two groups that correlate with their continents of origin. Considerable variation occurred among isolates previously classified as E. parva. Most isolates are placed into two closely related groups, but the remaining isolates, including some from human sources, are phylogenetically distinct and represent undescribed species. Strains ofB. dermatitidisare a sister species of E. parva. Paracoccidioides brasiliensisandHistoplasma capsulatumare ancestral to mostEmmonsiaisolates, and P. brasiliensis, which has no known teleomorph, falls within the Ajellomycesclade.

ABSTRACTSinus-orbital zygomycosis caused by Rhizomucor pusillusin a patient with acute myelogenous leukemia is described. Identification was achieved by sequencing of the internal transcribed spacer (ITS) regions of the rRNA gene and by expression of zygospores in mating. This report highlights the value of ITS sequencing as a diagnostic tool for the identification of R. pusillusand expands the understanding of infection types caused by this zygomycete.

ABSTRACTWe describe a case in which the Histoplasma capsulatumAccuProbe test displayed cross-reactivity with a respiratory isolate thought to be Histoplasmabut not morphologically consistent with H. capsulatum. The isolate was later identified as the Chrysosporiumanamorph of Nannizziopsis vriesiiby sequence analysis and phenotypic data.

ABSTRACTWe report a case of noninvasive pulmonary disease due to Chrysosporium zonatumin an immunocompetent male. The fungus colonized an existing tuberculous cavity and was isolated from transbronchial lavage fluid and from a percutaneous aspiration specimen. The disease was accompanied by the unusual feature of an allergic reaction. The fungus ball was successfully treated by intracavitary administration of amphotericin B. C. zonatumis the anamorph of the heterothallic ascomycete Uncinocarpus orissi, and the identity of the case isolate was verified by formation of ascospores in mating tests with reference isolates.