Abstracts
First human case of fungal keratitis caused by a putatively novel species of Lophotrichus.
Journal of Clinical Microbiology 2014; 52:4407-4411
Sue PK, Gurda GT, Lee R, Watkins T, Green R, Memon W, Milstone AM, Zelazny AM, Fahle GA, Pham TA, Gibas CF, Sutton DA, Wickes BL, Wiederhold NP, Zhang SX
Angioinvasive fungal infections (AFIs) are an important cause of morbidity and mortality among immunocompromised patients. However, clinicomicrobiological characteristics and treatment of many AFI agents remain poorly defined. We report the first human case of infection with Westerdykella dispersa, an emergent cause of AFI, which was successfully treated in a neutropenic pediatric patient.
Molecular characterization of reptile pathogens currently known as members of the Chrysosporium anamorph of Nannizziopsis vriesii (CANV) complex and relationship with some human-associated isolates.
Journal of Clinical Microbiology 2013; 51:3338-3357.
Sigler L, Hambleton S, Pare JA.
In recent years, the Chrysosporium anamorph of Nannizziopsis vriesii (CANV), Chrysosporium guarroi, Chrysosporium ophiodiicola and Chrysosporium species have been reported as the cause of dermal or deep lesions in reptiles. Infections are contagious, often fatal and affect both captive and free ranging animals. Forty nine CANV isolates from reptiles and six isolates from human sources were compared with Nannizziopsis vriesii based on cultural characteristics and DNA sequence data. Analyses of sequences of the internal transcribed spacer and small subunit of the nuclear ribosomal gene revealed the reptile pathogens and human isolates to belong in well supported clades corresponding to three lineages distinct from all other taxa within the family Onygenaceae of the Onygenales. One lineage represents the genus Nannizziopsis and comprises N. vriesii and seven 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 ophiodiicola occurring only on snakes, and Paranannizziopsis gen. nov. with three species from squamates and tuataras. The species newly described are Nannizziopsis dermatitidis, Nannizziopsis crocodili, Nannizziopsis barbata , Nannizziopsis infrequens, Nannizziopsis hominis, Nannizziopsis obscura, Paranannizziopsis australasiensis, Paranannizziopsis californiensis and Paranannizziopsis crustacea. Chrysosporium guarroi is transferred as Nannizziopsis guarroi. N. guarroi causes yellow fungus disease, a common infection in bearded dragons and green iguanas, and O. ophiodiicola is an emerging pathogen of captive and free-ranging snakes. Human-associated species were not recovered from reptiles and reptile-associated species were recovered only from reptiles, thereby mitigating zoonotic concerns.
Invasive sino-orbital mycosis in an aplastic anemia patient caused by Neosartorya laciniosa
Journal of Clinical Microbiology 2013; 51:1316-1319 [online 23 Jan 2013]
http://jcm.asm.org/cgi/content/abstract/51/4/1316?etoc
Malejczyk K, Sigler L, Gibas CFC, Smith SW
We report the first case of Neosartorya laciniosa invasive 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 benA and calmodulin genes.
Dermatitis and cellulitis in leopard geckos (Eublepharis macularius) caused by the Chrysosporium anamorph of Nannizziopsis vriesii.
Veterinary Pathology 2013; 50:585-589. [online Nov 2012] doi: 10.1177/0300985812465324.
Toplon DE, Terrell SP, Sigler L, Jacobson ER.
An epizootic of ulcerative to nodular ventral dermatitis was observed in a large breeding colony of 8-month to 5-year-old leopard geckos (Eublepharis macularius) of both sexes. Two representative mature male geckos were euthanized for diagnostic necropsy. The Chrysosporium anamorph of Nannizziopsis vriesii (CANV) was isolated from the skin lesions, and identification was confirmed by sequencing of the internal transcribed spacer region of the rRNA gene. Histopathology revealed multifocal to coalescing dermal and subcutaneous heterophilic granulomas that contained septate fungal hyphae. There was also multifocal epidermal hyperplasia with hyperkeratosis, and similar hyphae were present within the stratum corneum, occasionally with terminal chains of arthroconidia consistent with the CANV. In one case, there was focal extension of granulomatous inflammation into the underlying masseter muscle. This is the first report of dermatitis and cellulitis due to the CANV in leopard geckos.
Cytological, microbiological and therapeutic aspects of systemic infection in a dog caused by the fungus Phialosimplex caninus.
Medical Mycology Case Reports 2013; 2:32-36. [online 11 Jan 2013
http://dx.doi.org/10.1016/j.mmcr.2012.12.007
Sigler L, Hanselman B, Ruotsalo K, Tsui KG, Richardson S.
A seven-year-old immunocompetent dog presenting with lymphadenopathy, mesenteric masses and splenic nodules was diagnosed with Phialosimplex caninus infection. Cytology of a mesenteric mass aspirate demonstrated few intact cells but numerous variably sized fungal cells and rare hyphal fragments. The identity of the cultured fungus was confirmed by DNA sequencing. Itraconazole therapy improved clinical signs, but the fungus was reisolated at follow-up. P. caninus systemic infection should be suspected in dogs presenting with lymphadenopathy and splenomegaly.
Fungal myelitis caused by Phialosimplex caninus in an immunosuppressed dog.
Medical Mycology 2012; 50(5):509–512.
Armstrong P,
Sigler L, Sutton D, Grooters A, Hitt M.
A bone marrow infection caused by Phialosimplex caninus was diagnosed in a seven year-old female spayed Cocker Spaniel that was receiving prednisone for autoimmune hemolytic anemia. Histopathologic examination of a bone marrow core biopsy revealed clusters of oval to round yeast-like cells of varying shape and size and occasional irregular hyphae. Culture of a bone marrow aspirate sample yielded a mould initially suggestive of Paecilomyces inflatus or Sagenomella species but later determined to be P. caninus. The dog was treated with itraconazole and amphotericin B, and prednisone was continued at the lowest dose needed to control the hemolytic anemia. The patient died after 18 months of treatment. This is the first detailed clinical report of infection caused by P. caninus, a newly described fungus associated with disseminated disease in dogs.
Invasive Mycoleptodiscus fungal cellulitis and myositis.
Medical Mycology 2012; 50(7):740-745.
Koo S, Sutton DA, Yeh WW, Thompson EH,
Sigler L, Shearer JF, Hofstra DE, Wickes BL, Marty FM.
We report progressive necrotizing fungal cellulitis and myositis in the leg of a patient with glioblastoma multiforme treated with temozolomide and corticosteroids. While the morphologic appearance of the isolate and its ability to grow at temperatures greater than 32° C were suggestive of
Mycoleptodiscus indicus, some of the conidia were atypical for this species in that they had single septa and occasional lateral appendages. Furthermore, the isolate was different from
M. indicus based on the sequencing analysis of two rDNA regions. This is the first case of
Mycoleptodiscus invasive fungal disease in which the causative agent could not be resolved at the species level because of inconsistencies between morphological and molecular data.
Invasive Scopulariopsis brevicaulis infection in an immunocompromised patient and review of prior cases caused by Scopulariopsis and Microascus species.
Medical Mycology 2012; 50(6):561-569.
Iwen PC, Schutte SD, Florescu DF, Noel-Hurst RK,
Sigler L.
Scopulariopsis species and their Microascus teleomorphs are cosmopolitan fungi that are uncommonly associated with invasive disease. This report describes a case of fatal disseminated Scopulariopsis brevicaulis disease in a patient with diffuse large B cell lymphoma who underwent high-dose chemotherapy followed by a matched unrelated donor stem cell transplant. This case is compared with 32 prior cases of proven invasive Scopulariopsis (Microascus) infections reported in the literature. A focus of this report is the diagnostic methods utilized which included histopathology and culture with both micromorphologic and genotypic procedures employed to confirm the species identification.
Geosmithia argillacea: An emerging cause of invasive mycosis in human chronic granulomatous disease.
Clinical Infectious Diseases 2011; 52(6):e136-143.
De Ravin SS, Challipalli M, Anderson V, Shea YR, Marciano B, Hilligoss D, Marquesen M, Decastro R, Liu YC, Sutton DA, Wickes BL, Kammeyer PL,
Sigler L, Sullivan K, Kang EM, Malech HL, Holland SM, Zelazny AM.
BACKGROUND: Chronic granulomatous disease (CGD) is an inherited disorder of the nicotinamide adenine dinucleotide phosphate oxidase that leads to defective production of microbicidal superoxide and other oxidative radicals, resulting in increased susceptibility to invasive infections, especially those due to fungi. METHODS: Geosmithia argillacea was identified from cultured isolates by genomic sequencing of the internal transcribed spacer region. Isolates previously identified as Paecilomyces variotii, a filamentous fungus closely resembling G. argillacea, were also examined. RESULTS: We identified G. argillacea as the cause of invasive mycosis in 7 CGD patients. In 5 cases, the fungus had been previously identified morphologically as P. variotii. All patients had pulmonary lesions; 1 had disseminated lesions following inhalational pneumonia. Infections involved the chest wall and contiguous ribs in 2 patients and disseminated to the brain in 1 patient. Four patients with pneumonia underwent surgical intervention. All patients responded poorly to medical treatment, and 3 died. CONCLUSIONS: We report the first cases of invasive mycosis caused by G. argillacea in CGD patients. G. argillacea infections in CGD are often refractory and severe with a high fatality rate. Surgical intervention has been effective in some cases. G. argillacea is a previously underappreciated and frequently misidentified pathogen in CGD that should be excluded when P. variotii is identified morphologically.
Invasive Myceliophthora thermophila infection mimicking invasive aspergillosis in a neutropenic patient: a new cause of cross-reactivity with the Aspergillus galactomannan serum antigen assay.
Medical Mycology 2011; 49(8):883-886.
Morio F, Fraissinet F, Gastinne T, Pape PL, Delaunay J, Sigler L, Gibas CF, Miegeville M.
Myceliophthora thermophila is a thermophilic mould widely found in the environment but rarely responsible for human infections. We describe a case of invasive Myceliophthora thermophila infection mimicking invasive aspergillosis in a neutropenic patient with haematological malignancy. Cross-reactivity with Aspergillus galactomannan assay (GM) was demonstrated by repeated positive results and confirmed by cross-reaction between the fungal isolate and the GM assay. The patient was successfully treated with voriconazole. Potential GM cross-reactivity must be considered in future studies including patients categorized as having probable invasive aspergillosis using the GM as the only mycological criterion.
Computed tomography of granulomatous pneumonia in an American alligator (Alligator mississippiensis) associated with Metarhizium anisopliae.
Journal of Zoo and Wildlife Medicine 2011; 42(4):700-708.
Hall NH, Conley K, Berry C, Farina L, Sigler L, Wellehan JF Jr, Heard D.
An 18-yr-old, male, albino, American alligator (Alligator mississippiensis) was evaluated for decreased appetite and abnormal buoyancy. Computed tomography (CT) of the coelomic cavity showed multifocal mineral and soft tissue attenuating pulmonary masses consistent with pulmonary fungal granulomas. Additionally, multifocal areas of generalized, severe emphysema and pulmonary and pleural thickening were identified. The alligator was euthanized and necropsy revealed severe fungal pneumonia associated with oxalosis. Metarhizium anisopliae var. anisopliae was cultured from lung tissue and exhibited oxalate crystal formation in vitro. Crystals were identified as calcium oxalate monohydrate by X-ray powder defractometry. Fungal identification was based on morphology, including tissue sporulation, and DNA sequence analysis. This organism is typically thought of as an entomopathogen. Clinical signs of fungal pneumonia in nonavian reptiles are often inapparent until the disease is at an advanced stage, making antemortem diagnosis challenging. This case demonstrates the value of CT for pulmonary assessment and diagnosis of fungal pneumonia in the American alligator. Fungal infection with associated oxalosis should not be presumed to be aspergillosis.
Deep fungal dermatitis caused by the Chrysosporium anamorph of Nannizziopsis vriesii in captive coastal bearded dragons (Pogona barbata).
Australian Veterinary Journal 2011; 89(12): 515-519.
Johnson RSP, Sangster CR, Sigler L, Hambleton S, Paré JA.
Deep fungal dermatitis caused by the Chrysosporium anamorph of Nannizziopsis vriesii (CANV) was diagnosed in a group of coastal bearded dragons (Pogona barbata). The outbreak extended over a 6-month period, with four of six lizards from the same zoological outdoor enclosure succumbing to infection. A fifth case of dermatomycosis was identified in a pet lizard originally sourced from the wild. Diagnosis of infection with the CANV was based on similar clinical signs and histopathology in all animals and confirmed by culture and sequencing of the fungus from one animal. This is the first report of the CANV causing disease in a terrestrial reptile species in Australia and the first in the coastal bearded dragon.
Phialosimplex, a new anamorphic genus associated with infections in dogs and having phylogenetic affinity to the Trichocomaceae.
Medical Mycology 2010; 48:335-345
Sigler L, Sutton DA, Gibas CFC, Summerbell RC, Noel RK, Iwen PC.
Anamorphic members of the ascomycete family Trichocomaceae including Aspergillus, Penicillium, Paecilomyces, Geosmithia and Sagenomella have been reported from infections in canines. Six clinical isolates (five associated with infections in canines and one from a human source) demonstrated simple phialides producing conidia in long chains and were investigated for their potential relationship to Sagenomella chlamydospora, a known agent of canine disseminated mycosis. Phylogenetic analyses of internal transcribed spacer (ITS) and small subunit (SSU) region sequences revealed that all of the canine-associated isolates were distinct from Sagenomella species. The new anamorphic genus and species Phialosimplex caninus is described to accommodate the clinical isolates. Sagenomella chlamydospora and Sagenomella sclerotialis are transferred to the new genus as Phialosimplex chlamydosporus comb. nov. and Phialosimplex sclerotialis comb. nov.
Disseminated mycoses in veiled chameleons (Chamaeleo calyptratus) caused by Chamaeleomyces granulomatis, a new fungus related to Paecilomyces viridis.
Journal Clinical Microbiology 2010; 48:3182-3192. First published online Jul 21.
Sigler L,
Gibas CF, Kokotovica B, Bertelsen MF.
An 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 granulomatis gen. et sp. nov. is given as the name for the newly described fungus, and P. viridis is transferred to the new genus as Chamaeleomyces viridis comb. nov. Chamaeleomyces species 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. Chamaeleomyces species appear to be rare but aggressive pathogens of chameleons.
Xylogone ganodermophthora sp. nov., an ascomycetous pathogen causing yellow rot on cultivated mushroom Ganoderma lucidum in Korea.
Mycologia 2010; 102:1167–1184.
Kang, HJ,
Sigler L, Lee J,
Gibas CFC, Yun SH, Lee YW.
Yellow rot, caused by an ascomycetous fungus having a distinctive arthroconidial anamorph, is the most destructive disease of cultivated Ganoderma lucidum in 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. The YRP isolates formed a distinct subgroup within a clade that included X. sphaerospora, A. cuboidea and Scytalidium lignicola, the type species of Scytalidium, but the disposition of the clade within the Leotiomycetes was uncertain. We describe Xylogone ganodermophthora sp. nov. and Scytalidium ganodermophthorum sp. nov. for the teleomorph and anamorph of YRP, respectively. Arthrographis cuboidea is reclassified as Scytalidium cuboideum comb. nov. and the anamorph of X. sphaerospora is named Scytalidium sphaerosporum sp. nov. In pathogenicity tests, only X. ganodermophthora caused disease development in Ganoderma lucidum. Amplified fragment length polymorphism analyses showed that X. ganodermophthora populations from diseased fruiting bodies or from oak wood in Korea consisted of two clonal groups.
Fungal arthritis of the knee caused by Mycoleptodiscus indicus
Clinical Rheumatology 2010; 29:1061-1065
Dewar CL,
Sigler L
(
http://www.springerlink.com/openurl.asp?genre=article&id=doi:10.1007/s10067-010-1448-9.)
Mycoleptodiscus indicus is a recognized plant pathogen which has very rarely been reported as a cause of human infection. It is a tropical or subtropical fungus which is difficult to culture and identify from clinical specimens. This is the first report of septic arthritis with this fungus in a healthy Canadian male. The fungal infection was contracted on a vacation in Costa Rica, probably through direct inoculation through injured skin. The fungus was isolated from synovial fluid and identification was confirmed by DNA sequencing. There has only been one previous case of septic arthritis of the knee and one skin infection reported with this fungus; both cases involved immunocompromised hosts. Both septic arthritis patients required joint surgery and lavage to eradicate the fungus, however, only the immunocompromised patient required antifungal medications. In the future, it is very likely that the number of patients identified with
M. indicus infection will rise due to increasing awareness of this pathogen as well as increasing exposure. Many immunocompromised patients on anti-retroviral or biologic therapy are healthy enough to travel, thereby exposing themselves to exotic and infected plants which increase the risk of unusual fungal infections.
Pituitary cystadenoma, enterolipidosis, and cutaneous mycosis in an Everglades ratsnake (Elaphe obsoleta rossalleni).
Journal of Zoo Wildlife Medicine 2010; 41:538-541.
Dadone LI, Klaphake E, Garner MM, Schwahn D,
Sigler L, Trupkiewicz JG, Myers G, Barrie MT
An 11-yr-old captive-born male Everglades ratsnake (Elaphe obsoleta rosalleni) presented with dysecdysis, hyperkeratosis, and inappetance. Two skin biopsies demonstrated a diffuse hyperkeratosis with both a bacterial and fungal epidermitis. Fusarium oxysporum was cultured from both biopsies and considered an opportunistic infection rather than a primary pathogen. Medical management was unsuccessful, and the snake was euthanized. Histologic findings included a pituitary cystadenoma arising from the pars intermedia, severe intestinal lipidosis, generalized epidermal hyperkeratosis, and lesions consistent with sepsis. It is hypothesized that endocrine derangements from the pituitary tumor may have caused the skin and intestinal lesions.
Identification of a new species, Candida subhashii, as a cause of peritonitis
Medical Mycology 2009; 47:305-311.
Adam H, Groenewald M, Mohan S, Richardson S, Bunn U,
Gibas CFC, Poutanen S,
Sigler L
We report a case of fungal peritonitis from which a novel
Candida species was isolated. Phylogenetic analysis of DNA sequences from the internal transcribed spacer (ITS) region and the D1/D2 domains of the large subunit (LSU) rRNA gene show that the
Candida species is distinct from, but related to, the human pathogenic species,
C. parapsilosis, C. orthopsilosis, C. metapsilosis, C. tropicalis, C. albicans, and
C. dubliniensis.
Candida subhashii M. Groenewald, Sigler et Richardson sp. nov. is described.
Phialocephala urceolata , sp. nov., from a commercial, water-soluble heparin solution
Mycologia 2009; 101 136-141
Wang W, McGhee D,
Gibas, CFC, Tsuneda A, Currah RS.
Phialocephala urceolata sp. nov. was isolated from a black film that had developed on a watersoluble proprietary heparin solution (pH 2.5). Morphological and enzymatic characters, along with phylogenetic analyses of rDNA sequence data, indicated that the conidial fungus is closely related to species of Phialocephala known primarily as endophytes in the roots of vascular plants (e.g.
Acephala applanata, P. fortinii and
P. sphaeroides) or as associates of persistent plant organs such as the stems and needles of woody plants (e.g.
P. compacta, P. dimorphospora and
P. scopiformis).
Phialocephala urceolata is distinctive in having urn-shaped phialides that are sparsely distributed along the conidiophore axis, a slow growth rate in culture and in exhibiting a unique combination of reactions on enzymatic test media (i.e. it acidifies casamino acids medium and is gelatinase negative). Partial sequence data from the small subunit (SSU) rDNA indicated that
P. urceolata is among the Helotiales and close to the type species of
Phialocephala. Sequence data from the internal transcribed spacer (ITS) region places P. urceolata closest to P. sphaeroides. The source of this contaminant is unknown but its taxonomic relationship with other root endophytic species and its ability to produce polyphenol oxidases suggest that the natural habitat of this species is possibly woody plant tissues or soil enriched with lignocellulose.
Disseminated fungal infection in a renal transplant recipient involving Macrophomina phaseolina and Scytalidium dimidiatum: case report and review of taxonomic changes among medically important members of the Botryosphaeriaceae
Medical Mycology 2008; 6:285-292.
Tan DH, Sigler L, Gibas, CFC, Fong IW.
We report the first case of human infection with the fungal plant pathogen Macrophomina phaseolina in a Sri Lankan-born Canadian man following a renal transplant in India. The patient subsequently succumbed to invasive infection with Scytalidium dimidiatum. Molecular sequence analysis confirmed the identification of both fungi and revealed that they are related species within the ascomycete family Botryosphaeriaceae. We review the rationale for the recent reclassification of S. dimidiatum as Neoscytalidium dimidiatum and of Nattrassia mangiferae (formerly considered a synanamorph of S. dimidiatum) as Neofusicoccum mangiferae. This and other recent cases illustrate the potential for plant pathogenic fungi to cause invasive human diseases which are refractory to antifungal therapy.
Cryptosporiopsis species isolated from the roots of aspen in central Alberta: identification, morphology, and interactions with the host, in vitro.
Cananadian Journal of Botany 2007; 85: 1214–1226
Wang W, Tsuneda A,Gibas, CFC, Currah RS
Cryptosporiopsis Bubák & Kabát isolates were obtained for the first time from roots of apparently healthy aspen seedlings in Alberta. These isolates were similar in all the major morphological features previously used to separate Cryptosporiopsis species, but sequencing data of the ITS1-5.8S-ITS2 region indicated that they were separated into two groups, one belonging to Cryptosporiopsis ericae Sigler and the other to Cryptosporiopsis radicicola Kowalski & Bartnik. Scanning electron microscopy of ex-type cultures and selected isolates from aspen roots revealed that C. ericae and C. radicicola differed in morphogenesis and structure of conidiomata: those of C. ericae were either synnematous or sporodochial, whereas those of C. radicicola possessed a peridium-like mycelial envelope bearing amorphous adhesive material. Phialides in the hymenium of C. radicicola were also embedded in amorphous matrix material but such material was absent in C. ericae. Microscopic examination of artificially inoculated aspen roots indicated that both species are endophytes of the host. Hyphal penetration by C. ericae was only occasional and confined to the host epidermis, whereas C. radicicola was more aggressive and its hyphal ingress extended to the cortical region .
DNA and the classical way: Identification of medically important molds in the 21st century
Medical Mycology 2007; 45:475 - 490
Balajee SA, Sigler L, Brandt ME
The advent of the 21st century has seen significant advances in the methods and practices used for identification of medically important molds in the clinical microbiology laboratory. Historically, molds have been identified by using observations of colonial and microscopic morphology, along with tables, keys and textbook descriptions. This approach still has value for the identification of many fungal organisms, but requires expertise and can be problematic in determining a species identification that is timely and useful in the management of high-risk patients. For the increasing number of isolates that are uncommon, atypical, or unusual, DNA-based identification methods are being increasingly employed in many clinical laboratories. These methods include the commercially available GenProbe assay, methods based on the polymerase chain reaction such as single-step PCR, RAPD-PCR, rep-PCR, nested PCR, PCR-RFLP, PCR-EIA, and more recent microarray-based, Luminex technology-based, and real-time PCR-based methods. Great variation in assay complexity, targets, and detection methods can be found, and many of these methods have not been widely used or rigorously validated. The increasing availability of DNA sequencing chemistry has made comparative DNA sequence analysis an attractive alternative tool for fungal identification. DNA sequencing methodology can be purchased commercially or developed in-house; such methods display varying degrees of usefulness depending on the breadth and reliability of the databases used for comparison. The future success of sequencing-based approaches will depend on the choice of DNA target, the reliability of the result, and the availability of a validated sequence database for query and comparison. Future studies will be required to determine sequence homology breakpoints and to assess the accuracy of molecular-based species identification in various groups of medically important filamentous fungi. At this time, a polyphasic approach to identification that combines morphologic and molecular methods will ensure the greatest success in the management of patients with fungal infections.
Deep fungal dermatitis in three inland bearded dragons (Pogona vitticeps) caused by the Chrysosporium anamorph of Nannizziopsis vriesii.
Medical Mycology 2007; 45:291-296.
Bowman MR, Paré JA,
Sigler L, Naeser JP, Sladky KK, Hanley CS, Helmer P, Phillips LA, Brower A, Porter R
The
Chrysosporium anamorph of
Nannizziopsis vriesii (CANV), a keratinophilic fungus that naturally and experimentally causes severe and often fatal dermatitis in multiple reptile species, was isolated in pure culture from skin samples of three inland bearded dragons (
Pogona vitticeps) with deep granulomatous dermatomycosis. The first animal presented with a focal maxillary swelling involving the skin and gingiva. This lizard died while undergoing itraconazole and topical miconazole therapy. The second presented with focally extensive discoloration and thickening of the skin of the ventrum and was euthanized after 10 weeks of itraconazole therapy. A third lizard presented with hyperkeratotic exudative dermatitis on a markedly swollen forelimb. Amputation and itraconazole therapy resulted in a clinical cure. Histopathology of tissue biopsies in all cases demonstrated granulomatous dermatitis with intralesional hyphae morphologically consistent with those produced by the CANV. The second lizard also had granulomatous hepatitis with intralesional hyphae. Evidence in this report suggests that the CANV is the etiologic agent of an emerging condition in captive bearded dragons that has been called 'yellow fungus disease'.
Graphium basitruncatum fungemia in a patient with acute leukemia
Journal of Clinical Microbiology 2007; 45:1644-1647
Kumar D, Sigler L, Gibas CFC, Mohan S, Schuh A, Medeiros BC, Peckham K, Humar A.
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.
Mucor circinelloides was identified by molecular methods as a cause of primary cutaneous zygomycosis
Journal of Clinical Microbiology 2007; 45:636-640.
Iwen PC, Sigler L, Freifeld AG
A case of primary cutaneous zygomycosis caused by 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.
Ascoma development and phylogeny of an apothecioid dothideomycete, Catinella olivacea
American Journal of Botany 2007; 94:1890-1899
Greif MD,
Gibas CFC, Tsuneda A, Currah RS
Catinella olivacea is a discomycetous fungus often found fruiting within cavities in rotting logs. Because this habitat would lack the air currents upon which discomycete species normally rely for the dispersal of their forcibly ejected ascospores, we suspected an alternative disseminative strategy might be employed by this species. An examination of the development of the discomycetous ascomata in pure culture, on wood blocks, and on agar showed that the epithecium was gelatinous at maturity and entrapped released ascospores in a slimy mass. We interpreted this as an adaptation for ascospore disperal by arthropods. Developmental data also showed that C. olivacea was unusual among other discomycetes in the Helotiales (Leotiomycetes). For example, the ascoma developed from a stromatic mass of meristematically dividing cells and involved the formation of a uniloculate cavity within a structure better considered an ascostroma than an incipient apothecium. Furthermore, the ascus had a prominent ocular chamber and released its ascospores through a broad, bivalvate slit. These features, along with phylogenetic analyses of large subunit and small subunit rDNA, indicated that this unusual apothecial fungus is, surprisingly, more closely affiliated with the Dothideomycetes than the Leotiomycetes.
Monodictys arctica, a new hyphomycete from the roots of Saxifraga oppositifolia collected in the Canadian High Arctic
Mycotaxon 2006; 98:261-272
Day MJ, Gibas CFC, Fujimura KE, Egger KN, Currah RS
Monodictys arctica sp. nov. is described on the basis of nine isolates obtained from the roots of eight separate collections of Saxifraga oppositifolia from Ellesmere Island, Nunavut, Canada. Conidia are multicelled, smooth, darkly pigmented, and globose, oblong, ellipsoidal, or pyriform to irregularly shaped or dichotomously branched, consisting of a blastically produced basal cell and a distal proliferation of up to 24 cells arising from meristematic growth. Analyses of SSU and ITS sequences indicate the species is unique but has an affinity to the loculoascomycete taxon Leptosphaeria dryadophila.
Leptographium piriforme sp. nov., from a taxonomically diverse collection of arthropods collected in an aspen-dominated forest in western Canada
Mycologia 2006; 98 771-780.
Greif MD,
Gibas CFC, Currah RS
During a survey of fungi associated with arthropods collected in a southern boreal mixed-wood forest in Alberta we obtained 29 isolates of a unique species of Leptographium. This species displayed a distinct combination of characteristics, including curved conidia on short-stipitate conidiophores, a secondary micronematous conidial state, stalked pear-shaped cells and an optimal growth rate at 35 C, and is described as Leptographium piriforme sp. nov. The isolates were most similar morphologically to L. crassivaginatum, but ITS sequence comparisons indicate that our isolates cannot be assigned to this or any other sequenced species in the genus. Initial observations on the pear-shaped cells in feeding experiments with Sancassania berlesei show that these structures may act as a nutritional incentive for visiting arthropods. Most arthropods carrying this new species were caught in traps baited with dung which, in light of its optimum growth temperature, suggests a coprophilous phase in the life cycle of this species. Additional isolates from woody species typical of the survey area might clarify whether Leptographium piriforme in its forest habitat occurs as a plant pathogen or saprobe.
A new species of Cladophialophora (hyphomycetes) from boreal and montane bryophytes
Mycological Research 2006; 111: 106-116
Davey ML, Currah RS
During a survey of bryophilous fungi from boreal and montane habitats in central Alberta, a hitherto undescribed species of Cladophialophora was recovered from Polytrichum juniperinum, Aulacomnium palustre, and Sphagnum fuscum. On potato dextrose agar (PDA) colonies grew slowly, attaining a diameter of 25 mm after 30 d, were dark grey, velvety, radially sulcate, and convolute and cracked at the centre. Micronematous conidiophores gave rise to branched chains of small (1–2 _ 8–22 mm), cylindrical to fusiform conidia with truncate, swollen scars at each end. Phylogenies built on the ITS and ribosomal SSU regions indicate the isolates form a monophyletic clade within the family Herpotrichiellaceae (Chaetothyriales) that is composed of two geographically based groups, each with 99 % within-group sequence similarity and 97–98% between-group sequence similarity. Ateleomorph has not been found but would likely be similar to species of Capronia. In vitro inoculation of the isolates onto axenically grown P. juniperinum produced no discernible host symptoms, and host penetration could not be detected using light microscopy. The production of polyphenol oxidases by the fungus and the role of other Cladophialophora species as latent endophytes and saprobes suggest that a potential role for the fungus is the degradation of the polyphenol-rich cell walls of mosses. A dichotomous key to species of the genus Cladophialophora is provided.
Pathogenicity of the Chrysosporium anamorph of Nannizziopsis vriesii for veiled chameleons (Chamaeleo calyptratus)
Medical Mycology 2006; 44:25-31
Pare JA, Coyle KA,
Sigler L, Maas AK, Mitchell RL
Veiled chameleons (
Chamaeleo calyptratus) were experimentally challenged with the fungus
Chrysosporiumanamorph of
Nannizziopsis vriesii (CANV). Chameleons were exposed to conidia in their captive environment, or were inoculated by direct application of a conidial suspension inoculum on intact and on abraded skin. The CANV induced lesions in all experimental groups and was recovered from infected animals, fulfilling Koch's postulates and confirming that it may act as a primary fungal pathogen in this species of reptile. A breach in cutaneous integrity, as simulated by mild scarification, increased the risk of infection but was not required for the CANV to express pathogenicity. Initial hyphae proliferation occurred in the outer epidermal stratum corneum, with subsequent invasion of the deeper epidermal strata and dermis. A spectrum of lesions was observed ranging from liquefactive necrosis of the epidermis to granulomatous inflammation in the dermis. CANV dermatomycosis appears to be contagious and can readily spread within a reptile collection, either directly through contact with infective arthroconidia or indirectly via fomites. Dense tufts of arthroconidiating hyphae were demonstrated histologically on the skin surface of many animals that developed dermatomycosis, and these arthroconidia may act as infective propagules involved in the transfer of disease between reptiles.
Fatal cutaneous mycosis in tentacled snakes (Erpeton tentaculatum) caused by Chrysosporium anamorph of Nannizziopsis vriesii
Journal of Zoo & Wildlife Medicine 2005; 36:82-87.
Bertelsen MF, Crawshaw GJ, Sigler L, Smith DA
The fungus Chrysosporium anamorph of Nannizziopsis vriesii was identified as the cause of fatal, multifocal, heterophilic dermatitis in four freshwater aquatic captive-bred tentacled snakes (Erpeton tentaculatum). Pale, 1- to 4-mm focal lesions involving individual scales, occurred primarily on the head and dorsum. Histology showed multifocal coagulation necrosis of the epidermis, with marked heterophilic infiltration without involvement of the underlying dermis. Septate, irregularly branched hyphae, and clusters of 4- to 8- by 2- to 3-µm rod-shaped cells (arthroconidia) were present within the lesions and in a superficial crust. Failure to maintain an acidic environment was likely a predisposing factor in the development of these lesions.
False-positive Histoplasma capsulatum Gen-Probe chemiluminescent test caused by a Chrysosporium species.
Journal of Clinical Microbiology 2005; 43:1456-1458
Brandt ME, Gaunt D, Iqbal N, McClinton S, Hambleton S,
Sigler L
We describe a case in which the
Histoplasma capsulatum AccuProbe test displayed cross-reactivity with a respiratory isolate thought to be
Histoplasma but not morphologically consistent with
H .
capsulatum . The isolate was later identified as the
Chrysosporium anamorph of
Nannizziopsis vriesii by sequence analysis and phenotypic data.
Meliniomyces, a new anamorph genus for root-associated fungi with phylogenetic affinities to Rhizoscyphus ericae ( = Hymenoscyphus ericae), Leotiomycetes
Studies in Mycology 2005; 53:1-27
Hambleton S,
Sigler L
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 myrtilloides or
V. vitis-idaea , isolates did not form typical ericoid mycorrhizas. Additional isolates obtained from roots of the
Orchidaceae ,
Pinaceae ,
Betulaceae and
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 ericae aggregate” which includes
H. ericae (
Leotiomycetes ), many unnamed taxa and
Cadophora finlandica . The new genus and species
Meliniomyces variabilis is 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. variabilis is 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. ericae aggregate” includes several other well-supported clades putatively named as
Meliniomyces species. Representative strains were examined morphologically for two of these species, described as
M. vraolstadiae and
M. bicolor . Both include ectomycorrhizal mycobionts of the “
Piceirhiza bicolorata ” morphotype.
Rhizoscyphus ericae is accepted as the appropriate name for
H. ericae .
Molecular identification of Rhizomucor pusillus as a cause of sinus-orbital zygomycosis in a patient with acute myelogenous leukemia
Journal of Clinical Microbiology 2005; 43:5819-5821
Iwen PC, Freifeld AG, Sigler L, Tarantolo,SR
Sinus-orbital zygomycosis caused by Rhizomucor pusillus in 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. pusillus and expands the understanding of infection types caused by this zygomycete.
Peritonitis due to Curvularia inaequalis in an elderly patient undergoing peritoneal dialysis and a review of six cases of peritonitis associated with other Curvularia spp
Journal of Clinical Microbiology 2005; 43: 4288–4292
Pimentel JD, Mahade van K, Woodgyer A, Sigler L, Gibas C, Harris OC, Lupino M, Athan E
Fungal peritonitis due to Curvularia species 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 Curvularia species.
Two new Cryptosporiopsis species from roots of ericaceous hosts
Studies in Mycology 2005; 53:53-62
Sigler L, Allan T, Lim SR, Berch S, Berbee M
Cryptosporiopsis species are anamorphs of ascomycetes in the genera Pezicula and 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 Pezicula and, when regrown, revealed conidiomata and conidia typical of Cryptosporiopsis species. Cultural and molecular data allowed for the ecognition of the new species,Cryptosporiopsis ericae and C. brunnea .
Utility of a cultural method for identification of the ericoid mycobiont Oidiodendron maius confirmed by ITS sequence analysis
Studies in Mycology 2005; 53:61-72
Sigler L, Gibas CFC
A simple cultural method was investigated for its reliability in distinguishing the ericoid mycobiontOidiodendron maius 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. maius and its close relative O. citrinum expressed 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. maius with O. citrinum , O. griseum with O. flavum , and O. truncatum as 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. fuscum is here regarded as a distinct taxon.
Mating patterns and ITS sequences distinguish the sclerotial species Arachnomyces glareosus sp. nov. and Onychocola sclerotica
Studies in Mycology 2004; 50:525-531
Gibas CFC, Sigler L, Currah RS
Mating patterns among twelve strains of Onychocola sclerotica demonstrated that the ex-type was genetically distinct from all other strains. Ten strains, when crossed, produced an Arachnomyces state. A re-examination of morphology and an analysis of nuclear ribosomal internal transcribed spacer (ITS) region sequences supported recognizing the interfertile strains and one infertile strain as a species distinct from O. sclerotica. Arachnomyces glareosus (anamorph O. glareosa) sp. nov. and O. sclerotica are similar in producing a sclerotial synanamorph but they are phylogenetically distinct.
Acremonium exuviarum sp. nov., a lizard-associated fungus with affinity to Emericellopsis.
Studies in Mycology 2004; 50:409-413.
Sigler L, Zuccaro A, Summerbell RC, Mitchell JI, Paré JA
In a survey of cycloheximide-tolerant fungi growing from shed reptile skins, an Acremonium-like fungus was isolated that was distinctive in producing relatively large conidia in chains from phialides that tended to collapse after forming a number of conidia. Phylogenetic analysis based on ribosomal internal transcribed spacer and ß-tubulin sequences revealed that the isolate represented an undescribed and relatively phylogenetically isolated member of the clade containing the teleomorph genus Emericellopsis van Beyma as well as related anamorphs in Acremonium Link and Stanjemonium W. Gams, O’Donnell, Schoers & M. Chr. The species is here described as Acremonium exuviarum sp. nov. It has been isolated only on a single occasion and its ecology is unknown. Discovery of such new species in the pharmaceutically important Emericellopsis clade is potentially of practical significance.
Disseminated Beauveria bassiana infection in a patient with acute lymphoblastic leukemia.
Journal of Clinical Microbiology 2004; 42:5412-5414
Tucker DL, Beresford CH, Sigler L, Rogers K
We describe a case of disseminated Beauveria bassiana infection in a patient with acute lymphoblastic leukemia. Her infection was successfully treated with amphotericin B and itraconazole. B. bassiana is rarely reported as a human pathogen. It is commonly found in soil and because of its pathogenicity to many insect species is incorporated into several pesticides.
The Ajellomycetaceae, a new family of vertebrate-associated Onygenales.
Mycologia 2004; 96:811-820.
Untereiner WA, Scott JA, Naveau FA, Sigler L, Bachewich J, Angus A
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, Emmonsia and Histoplasma) andParacoccidioides 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.
Culture collections in Canada: perspectives and problems.
Canadian Journal of Plant Pathology 2004; 26:39-47
Sigler L
Culture collections are custodians of microbial resources of vital importance to science and society. These facilities are essential in enabling contemporary and future research in basic and applied sciences, and in integrating more than 75 years of records on Canadian microbial diversity. Culture collections often carry on because of dedicated efforts of key individuals. However, they become vulnerable to loss or dismantling when individuals retire or shift research direction in response to program reorganization or loss of funding. The need for conservation of, and long-term access to, microbial resources has long been recognized, and since 1962, six workshops have been held to address concerns about their future. In 1988, a report by the Task Force on the Status of Culture Collections in Canada made several recommendations. Key among these were that (1) specialized collections of strategic importance be supported, (2) an advisory committee be established to include members from different sectors of the scientific community, (3) government agencies allow user fees to be charged for access to collections, which would then be used for operational support, (4) the Natural Sciences and Engineering Research Council of Canada expand the infrastructure program to fund culture collections, and (5) technologies for improved access to vital data on strain history and properties be developed. Follow-up meetings resulted in a recommendation that an expert committee on plant and microbial genetic resources be established under the Canadian Agricultural Research Council. Although these activities resulted in increased recognition and support for some collections, in general, the situation of Canadian collections is no better, and is probably more dire, than in 1988. A national strategy is urgently needed to ensure the long-term care of valuable microbial genetic material.
Allergic fungal sinusitis associated with Trichoderma longibrachiatum.
Journal of Clinical Microbiology 2003; 41:5333-5336
Tang P, Mohan S, Sigler L, Witterick I, Summerbell R, Campbell I, Mazzulli T
We describe allergic fungal sinusitis caused by Trichoderma longibrachiatum in 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 Trichoderma was 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.
Survey for the Chrysosporium anamorph of Nannizziopsis vriesii on the skin of healthy captive squamates reptiles and notes on their cutaneous fungal mycobiota.
Journal of Herpetological Medicine and Surgery 2003; 13:10-15
Paré JA, Sigler L, Rypien KL, Gibas CFC
The Chrysosporium anamorph 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 setae. Fungi belonging to 50 genera were identified from 127 reptiles:Aspergillus spp., Penicillium spp., and Paecilomyces lilacinus were 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.
Mycorrhizal and root endophytic fungi of containerized Picea glauca seedlings assessed by rDNA sequence analysis
Microbial Ecology 2003; 45: 128-136
Kernaghan G, Sigler L, Khasa D
Fungi colonizing fine roots of containerized Picea glauca seedlings were assessed in four large conifer nurseries in northern Alberta. PCR amplification of fungal rDNA (internal transcribed spacer and a portion of the 5' end of the large subunit gene) from random samples of fine feeder roots gave between 1 and 4 amplicons per seedling. Amplicons were either separated by electrophoresis and sequenced directly, or cloned and sequenced. The resulting sequences were compared to sequences obtained from cultures established from seedling roots and from Gen-Bank by maximum parsimony analysis. ITS sequences formed 11 distinct clades, each including at least one reference sequence. The ectomycorrhizal basidiomycetes Thelephora americana and Amphinema byssoides were dominant, whereas ascomycetes were less common. Fungi with sequences similar to members of the Heleotiales which form ericoid mycorrhizas were also present. Correspondence analysis revealed strong positive and negative associations among fungal taxa as well as an influence of applied fertilizer level on fungal diversity and species composition.
Phylogeny of the genus Arachnomyces and its anamorphs and the establishment of Arachnomycetales, a new eurotiomycete order in the Ascomycota
Studies in Mycology 2002; 47: 131-139
Gibas CFC, Sigler L, Summerbell RC, Currah RS
Arachnomyces is a genus of cleistothecial ascomycetes that has morphological similarities to the Onygenaceae and the Gymnoascaceae but is not accommodated well in either taxon. The phylogeny of the genus and its related anamorphs was studied using nuclear SSU rDNA gene sequences. Partial sequences were determined from ex-type cultures representing A. minimus, A. nodosetosus (anamorph Onychocola canadensis), A. kanei (anamorph O. kanei) and A. gracilis (anamorph Malbranchea sp.) and aligned together with published sequences of onygenalean and other ascomycetes. Phylogenetic analysis based on maximum parsimony showed that Arachnomyces is monophyletic, that it includes the hyphomyceteMalbranchea sclerotica, and it forms a distinct lineage within the Eurotiomycetes. Based on molecular and morphological data, we propose the new order Arachnomycetales and a new family Arachnomycetaceae. All known anamorphs in this lineage are arthroconidial and have been placed either in Onychocola (A. nodosetosus, A. kanei) or in Malbranchea (A. gracilis). Onychocola is considered appropriate for disposition of the arthroconidial states of Arachnomyces and thus Malbranchea sclerotica and the unnamed anamorph of A. gracilis are redisposed as Onychocola sclerotica comb. nov. and O. gracilis sp. nov.
Chlamydosauromyces punctatus gen. & sp. nov. (Onygenaceae) from the skin of a lizard
Studies in Mycology 2002; 47: 123-129
Sigler L, Hambleton S, Paré JA
Chlamydosauromyces punctatus is described for an ascomycete producing punctate, rimmed ascospores within ascomata composed of narrow, thin-walled, branched hyphae and an anamorph of alternate arthroconidia. It is known from a single collection obtained from shed skin of a frilled lizard. DNA sequences from the small subunit (SSU) region of the nuclear ribosomal gene were obtained from the lizard isolate and two other taxa and compared with homologous sequences of onygenalean fungi obtained from GenBank. Phylogenetic analysis supports the inclusion of the genus Chlamydosauromyces within theOnygenaceae. Results also supported the separation of Arachniotus ruber from Kraurogymnocarpa trochleospora (Pseudoarachniotus trochleosporus), a species that at one time was considered synonymous.
Auxarthron teleomorphs for Malbranchea filamentosa and Malbranchea albolutea and relationships within Auxarthron
Studies in Mycology 2002; 47: 111-122
Sigler L, Hambleton S, Flis AL, Paré JA
Malbranchea filamentosa and M. albolutea were reexamined and connected to teleomorphs in the genus Auxarthron. DNA sequences were obtained from the small subunit (SSU) and internal transcribed spacer (ITS) regions of the nuclear ribosomal rRNA gene to evaluate conspecificity of the M. filamentosa isolates and to evaluate relationships among taxa within the Auxarthron clade. Five of eight isolates putatively identified as M. filamentosa produced fertile ascomata in matings and with F1 progeny. Auxarthron filamentosum sp. nov. is described for the teleomorph. Three nonmating isolates appeared conspecific based on morphology, but one was excluded based on sequence divergence. An Auxarthron teleomorph was described for M. albolutea in 1976, but not named because of uncertainty about its distinction from A. thaxteri and A. umbrinum. Auxarthron alboluteum sp. nov. is shown to be phylogenetically distinct. Phylogenetic analysis based on newly derived sequences showed that members of the genus Auxarthronand Malbranchea dendritica formed a strongly supported monophyletic group. In the ITS analysis, most species were placed into two well supported clades that correlated with the shape of the ascospores; the position of the type species A. californiense, was not clearly resolved. Differences were found between newly derived SSU sequences and those on deposit in GenBank for the same strains. After re-evaluation of the strains, the following sequences are considered to be incorrect: M. albolutea L28063 (UAMH 2846), A. zuffianum L28062 (UAMH 4098), M. dendritica L28064 (UAMH 2731) and M. filamentosa L28065 (UAMH 4097). The sequence for U29389 is correct for Malbranchea albolutea not M. dendritica as stated in the original GenBank record. The problems with these sequences were not uncovered in prior published analyses because insufficient representatives of Auxarthron species were sampled.
Arachnomyces kanei (anamorph Onychocola kanei) sp. nov., from human nails.
Medical Mycology 2002; 40:573–580
Gibas CFC, Sigler L, Summerbel RC, Hofstader SLR, Gupta AK
Five isolates of a slow-growing cycloheximide resistant hyphomycetous fungus were obtained from nail specimens and investigated for their relationship to Onychocola canadensis (teleomorphArachnomyces nodosetosus), a known agent of onychomycosis. In one patient diagnosed with superficial white onychomycosis, etiology was confirmed by a nail sample showing atypical filaments in direct microscopy, and by a follow-up specimen yielding cultures of the same fungus. A case of mixed infection with Aspergillus sydowii was also confirmed after examination of cultures grown from three successive microscopic-positive hallux nail specimens. For other isolates, etiological significance could not be confirmed by repeat sampling or results of direct microscopy were negative or unknown. Mating experiments yielded setose ascomata containing smooth oblate ascospores typical of Arachnomyces species. Phylogenetic analysis of ITS 2 region sequences support the conspecificity of the isolates and their placement within the genus. A. kanei sp. nov. (anamorph O. kanei sp. nov.) is described.
Four new Penicillium species having Thysanophora-like melanized conidiophores
Mycolological Research 2002; 106(9): 1109-1118.
Peterson SW, Sigler L
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.
Chrysosporium anamorph of Nannizziopsis vriesii associated with fatal cutaneous mycoses in the saltwater crocodile (Crocodylus porosus)
Medical Mycology 2002; 40: 143–151
Thomas AD, Sigler L, Peucker S, Norton JH, Nielan A
The Chrysosporium anamorph of Nannizziopsis vriesii, recently identified as the cause of cutaneous infections in chameleons and brown tree snakes, was associated with skin infections and deaths in salt-water crocodile (Crocodylus porosus) hatchlings on two separate occasions 3 years apart. In all, 48 animals died from the infection. All hatchlings came from the same farm in northern Queensland, Australia.
Pulmonary colonization by Chrysosporium zonatum associated with allergic inflammation in an immunocompetent subject
Journal of Clinical Microbiology 2002; 40: 1113–1115
Hayashi S, Naitoh K, Matsubara S, Nakahara Y, Nagasawa Z, Tanabe I, Kusaba K, Tadano J, Nishimura K, Sigler L
We report a case of noninvasive pulmonary disease due to Chrysosporium zonatum in 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. zonatum is the anamorph of the heterothallic ascomyceteUncinocarpus orissi, and the identity of the case isolate was verified by formation of ascospores in mating tests with reference isolates.
Thelebolus microsporus mycelial mats in the trachea of wild brown skua (Catharacta antarctica lonnbergi) and south polar skua (C. maccormicki) carcasses
Journal of Wildlife Diseases 2002; 38: 443–447
Leotta GA, Paré JA, Sigler L, Montalti D, Vigo G, Petruccelli M, Reinoso EH
Sixteen brown skuas (Catharacta antarctica lonnbergi) and seven South Polar skuas (C. maccormicki) were found dead near Boekella Lake, Hope Bay, Antarctica, in February 1997. Postmortem examination revealed conspicuous caseous, deep yellow fungal/mycelial mats or cores in the trachea of nine of 19 carcasses that were examined. These mycelial cores, highly suggestive of aspergillomas, completely occluded the tracheal lumen in four of these nine carcasses. Thelebolus microsporus, a psychrophilic ascomycetous fungus commonly isolated from skua dung and skua nesting material, was isolated in pure culture from these tracheal plugs. Awareness of pseudolesions resulting from Thelebolus microsporus profuse postmortem growth in the trachea of dead skuas will minimize potential confusion with aspergillosis when investigating causes of epornithics in Antarctica.
Use of holomorph characters to delimit Microascus nidicola and M. soppii sp. nov., with notes on the genus Pithoascus.
Mycologia 2002; 94(2): 362–369
Abbott SP, Lumley TC, Sigler L
Several isolates of a perithecial microascaceous ascomycete having falcate ascospores and a Scopulariopsis anamorph 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. soppii is described. The anamorph is S. soppii. Single ascospore isolates derived from M. nidicola demonstrated homothallism and lacked an anamorph.Scopulariopsis flava (basionym Acaulium flavum) is considered a nomen dubium. Generic concepts ofPithoascus are evaluated and the genus is treated as a synonym of Microascus. Pithoascus stoveri is transferred as M. stoveri comb. nov.
Effect of fertilization on seedling growth and ectomycorrhizal development of conifer seedlings in container-grown and bare-root nurseries.
New Forests 2001; 22:179-197
Khasa PD, Sigler L, Chakravarty P, Dancik BP, Erickson L, McCurdy D
Effect of three levels of fertilizer on the growth and ectomycorrhizal colonization by six species of ectomycorrhizal fungi (Hebeloma longicaudum, Laccaria bicolor, Paxillus involutus, Pisolithus tinctorius,Rhizopogon vinicolor and Suillus tomentosus) on three species of containerized-grown conifer seedlings (Pinus contorta, Picea glauca, and Picea mariana) and two species of bare-root conifer seedlings (Pinus sylvestris and Larix sibirica) was studied. Growth of the seedlings in both container-grown and bare-root nurseries increased as the levels of fertilizer increased. For better seedling growth and environmental quality it may be possible to reduce the level of fertilizers up to 33% by using selected mycorrhizal fungi. Ectomycorrhizal colonization in all seedlings was not affected by fertilizer levels. Hebeloma longicaudum, L. bicolor, P. involutus, and P. tinctorius formed well-developed ectomycorrhizae, whereas ectomycorrhizal development by R. vinicolor and S. tomentosus was poor. Native mycorrhizal fungi colonized non-inoculated control seedlings; however, their colonization was always lower than with inoculated fungi.
High mortality in a large-scale zebrafish colony (Brachydanio rerio Hamilton & Buchanan, 1822) associated with Lecythophora mutabilis (van Beyma) W. Gams & McGinnis.
Comparative Medicine 2001; 51(4):361-8.
Dykstra MJ, Astrofsky KM, Schrenzel MD, Fox JG, Bullis RA, Farrington S,
Sigler L, Rinaldi MG, McGinnis MR
Zebrafish (
Brachydanio rerio) have become an important model system for studying vertebrate embryonic development and gene function through manipulation of genotype and characterization of resultant phenotypes. An established research zebrafish colony without substantial disease problems for more than 7 years of operation began experiencing appreciable mortalities in November of 1997. Young fish (fry), from five to 24 days after hatching, spontaneously developed elongate strands of organic material protruding from the mouth, operculum, and anal pore, leading workers in the laboratory to describe the infected fish as "bearded." Unlike typical freshwater fish fungal infections, the skin surface did not have evidence of fungal colonization. The disease was associated with progressive lethargy, reduced feeding, and subsequent mortality. From 10 to 100% of the fry in a given tank were affected. Initial examination indicated that the biofilm around the head of affected fry consisted of bundles of septate fungal hyphae, large numbers of mixed bacterial populations, and protozoans. Environmental samples of air and water in the laboratory were obtained to ascertain the source of the infective agent and to isolate and identify the fungus. A fungus identified as
Lecythophora mutabilis was isolated repeatedly from infected fish and water samples from infected fish tanks, and from the main laboratory water supply tanks, but not from laboratory air. Some biofilm beards on fish were found to consist of relatively pure bacterial populations, and beards on occasional fish examined in the later part of the study consisted of hyphae and spores of the oomycete genus
Aphanomyces.
Lecythophora mutabilis did not invade tissues; however, elimination of the epizootic correlated with reduction in the number of
L. mutabilis conidia in the water following modification of the laboratory water system by use of new filtration and sterilization systems. We conclude that the dense hyphal strands of L. mutabilis composing the predominant biofilm type, along with mixed bacteria and protozoa, contributed to the die-off in young fry by occluding the oral cavity and/or gills, leading to starvation and/or asphyxiation
Heterothallism in the Microascaceae demonstrated by three species in the Scopulariopsis brevicaulis series.
Mycologia 2001; 93 (6): 1211-1220
Abbott SP,
Sigler L
Scopulariopsis anamorphs 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 brevicaulis was 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. fusca and
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 niger is 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.
White grain mycetoma caused by a Cylindrocarpon sp. in India
Journal of Clinical Microbiology 2000; 38:4288-4291.
Hemeshettar BM, Siddaramappa B, Padhye AA, Sigler L, Chandler FW
We describe a case of white grain eumycetoma of the foot of an Indian male caused by a slow-growing, poorly sporulating fungus that does not match any known agent of this infection. Histologic examination of a biopsy tissue specimen showed oval, lobular, white granules composed of hyaline, septate hyphae, and thick-walled chlamydospores. Culture of granules from a draining sinus yielded compact, very-slow-growing, poorly sporulating colonies producing a strong reddish brown pigment that diffused into the medium. The fungus was identified as a Cylindrocarpon sp. based on the development of rare cylindrical conidia borne from solitary phialides lacking collarettes, in addition to chlamydospores formed singly or in short chains.
New species and records of saprophytic ascomycetes (Myxotrichaceae) from decaying logs in the boreal forest.
Mycoscience 2000; 41:487-494
Sigler L, Lumley TC, Currah RS
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 Gymnostellatospora and Pseudogymnoascus. In addition to new reports of G. japonica, G. frigida and P.roseus, two new species are described. G. canadensis sp. nov. resembles G. japonica but differs in producing brown ascomata and in the formation of an arthroconidial anamorph. G. subnuda sp. nov. is distinct in lacking differentiated peridial hyphae. G. alpina was not found in decayed wood but is reviewed based on extralimital material. A dichotomous key to the five species of Gymnostellatospora is provided.
Acrophialophora fusispora brain abscess in a child with acute lymphoblastic leukemia. Review of cases and taxonomy.
Journal of Clinical Microbiology 2000; 38: 4569-4576.
Al-Mohsen IZ, Sutton DA, Sigler L, Almodovar E, Mahboub N, Frayha H, Al-Hajjar S, Rinaldi MG, Walsh T
A 12-year-old girl with acute lymphoblastic leukemia was referred to King Faisal Specialist Hospital and Research Center. The diagnosis without central nervous system (CNS) involvement was confirmed on admission, and chemotherapy was initiated according to the Children Cancer Group (CCG) 1882 protocol for high-risk-group leukemia. During neutropenia amphotericin B (AMB) (1 mg/kg of body weight/day) was initiated for presumed fungal infection when a computed tomography (CT) scan of the chest revealed multiple nodular densities. After 3 weeks of AMB therapy, a follow-up chest CT revealed progression of the pulmonary nodules. The patient subsequently suffered a seizure, and a CT scan of the brain was consistent with infarction or hemorrhage. Because of progression of pulmonary lesions while receiving AMB, antifungal therapy was changed to liposomal AMB (LAMB) (6 mg/kg/day). Despite 26 days of LAMB, the patient continued to have intermittent fever, and CT and magnetic resonance imaging of the brain demonstrated findings consistent with a brain abscess. Aspiration of brain abscess was performed and the Gomori methenamine silver stain was positive for hyphal elements. Culture of this material grewAcrophialophora fusispora. Lung biopsy showed necrotizing fungal pneumonia with negative culture. The dosage of LAMB was increased, and itraconazole (ITRA) was added; subsequently LAMB was discontinued and therapy was continued with ITRA alone. The patient demonstrated clinical and radiological improvement. In vitro, the isolate was susceptible to low concentrations of AMB and ITRA.
A. fusispora is a thermotolerant, fast-growing fungus with neurotropic potential. We report the first case of human infection involving the CNS. Acrophialophora resembles Paecilomyces but differs in having colonies that become dark and in the development of phialides along the sides or at the tips of echinulate brown conidiophores. Conidia are borne in long chains and are smooth or ornamented with fine-to-coarse echinulations, sometimes in spiral bands. The taxonomy of the genus Acrophialophora is reviewed, andAcrophialophora nainiana and Acrophialophora levis are considered as synonyms of A. fusispora .
Medical management of Beauveria bassiana keratitis.
Cornea 2000; 19:405-406
Kisla TA, Cu-Unjieng A, Sigler L, Sugar J
PURPOSE: To describe a case of Beauveria bassiana keratitis and to discuss the management of this rare condition. METHODS: An 82-year-old woman underwent surgical repair of a graft wound dehiscence. Seven months later, shortly after the removal of sutures, the patient developed a fungal keratitis. B. bassiana was identified as the infecting organism. The patient was treated with topical natamycin and oral fluconazole. RESULTS: Following antifungal therapy, the corneal ulcer was eradicated, but the patient underwent repeat penetrating keratoplasty for decreased vision due to corneal edema. The graft remains clear and visual acuity is substantially improved. CONCLUSION: The medical management of B. bassianakeratitis has previously been unsuccessful. The use of topical natamycin combined with oral fluconazole in the management of this case is discussed.
Pulmonary infection caused by Gymnascella hyalinospora in a patient with acute myelogenous leukemia.
Journal of Clinical Microbiology 2000;38:375-381
Iwen PC, Sigler L, Tarantolo S, Sutton DA, Rinaldi MG, Lackner RP, McCarthy DI, Heinrichs SH
We report the first case of invasive pulmonary infection caused by the thermotolerant ascomycetous fungus Gymnascella hyalinospora in 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 wasdischarged in stable condition with an absolute neutrophil count of 100 cells per microliter. 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 dermatitidis Culture 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 and5-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. hyalinospora based on its formation of oblate,smooth-walled ascospores within yellow or yellow-green tufts of aerial hyphae on sporulation media. Repeat testing with the Blastomyces probe demonstrated false-positive results with the case isolate and a reference isolate of G. 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.
New records from India and redescription of Corynascus thermophilus and its anamorph Myceliophthora fergusii
Mycotaxon 1998; 68:185-192
Sigler L, Aneja KR, Kumar R, Maheshwari R, Shukla RV
Studies of Indian thermophilic fungi have yielded several isolations of Myceliophthora fergusii, anamorph of the heterothallic ascomycete Corynascus thermophilus (Thielaviaceae). Identification was confirmed by matings. Chrysosporium fermentotritici is a synonym of M. fergusii. Studies of soil and decomposing jute in India recovered isolates of a rapid growing thermophilic conidial fungus initially thought to be a new species of Chrysosporium or Sporotrichum. The isolates were shown to be Corynascus thermophilus(Fergus & Sinden) Klopotek by mating studies. However because their identification had proven elusive to the authors and to several other investigators, we consider it appropriate to briefly redescribe the species, to review the taxonomic history, and to report the new records. The key features which distinguish the anamorph Myceliophthora fergusii (Klopotek) Oorschot from M. thermophila (Apinis) Oorschot are noted.
Cutaneous mycoses in chameleons caused by the Chrysosporium anamorph of Nannizziopsis vriesii (Apinis) Currah
J. Zoo Wildlife Medicine 1997; 28(4):443-453
Paré JA, Sigler L, Hunter DB, Summerbell RC, Smith DA, Machin KL
A dermatophyte-like fungus was isolated from skin biopsies of three different species of captive chameleon in which fungal elements had been observed by histological examination. An adult Parson's chameleon (Chamaeleo parsonii) presented with vesicles that became crusty brown lesions on the limbs and body. Skin biopsies revealed fungal hyphae in the affected epidermis and in underlying dermis. The lesions regressed fully after oral administration of itraconazole. An adult jewel chameleon (Chamaeleo lateralis) from the same private collection presented with localized black skin lesions and died while being treated with itraconazole. A pulmonary granuloma was also present in this chameleon at autopsy. Cultures obtained from skin and lung lesions yielded the same fungus. A third isolate was obtained from a skin biopsy of a Jackson’s chameleon (Chamaeleo jacksoni) with deep ulcerative cutaneous lesions located at the base of the tail. The fungus, in all three cases, has been identified as the Chrysosporium anamorph ofNannizziopsis vriesii, a poorly known ascomycetous species recorded previously from the skin of lizard and from soil, on the basis of its keratinolytic activity, resistance to cycloheximide, strongly restricted growth at 37 C, formation of clavate or pyriform single celled or two celled aleurioconidia, alternate and fission arthroconidia.
Molecular genetic variation in Emmonsia crescens and E. parva, etiologic agents of adiaspiromycosis, and their phylogenetic relationship to Blastomyces dermatitidis (Ajellomyces dermatitidis) and other systemic fungal pathogens.
Journal of Clinical Microbiology 1998; 36:2918-2925.
Peterson SW, Sigler L
Emmonsia crescens, an agent of adiaspiromycosis, Blastomyces dermatitidis, the agent of blastomycosis, and Histoplasma capsulatum, the agent of histoplasmosis, are known to form meiotic (sexual) stages in the ascomycete genus Ajellomyces (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 putativeAjellomyces clade, large subunit ribosomal and ITS region DNA sequences were determined from PCR amplified DNA fragments. Sequences were analyzed phylogenetically to evaluate the genetic variation within the genus Emmonsia and evolutionary relationships to other taxa. Emmonsia crescens and E. parvaare distinct species. Emmonsia crescens isolates 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 of B. dermatitidis are a sister species of E. parva. Paracoccidioides brasiliensis and Histoplasma capsulatum are ancestral to mostEmmonsia isolates, and P. brasiliensis, which has no known teleomorph, falls within the Ajellomyces clade.
Utility and features of a customized PC Windows-based relational database for managing microbial strain data
Journal of Industrial Microbiology and Biotechnology 1998; 20:86-89
Sigler L, Flis AL
In addition to storing microbes, culture collections in industry, government or universities manage a vast and continuously expanding library of information on strain history and properties. An efficient and cost-effective computer database system is required for entering, analyzing and searching these data. This report describes the utility and features of a comprehensive data base which consists of a commercially available relational database system combined with customized screens for data entry, viewing and report generation. The application was developed using Microsoft Access and Visual Basic to operate in the Windows environment on a local area network.
Microascus brevicaulis sp. nov., the teleomorph of Scopulariopsis brevicaulis, supports placement of Scopulariopsis with the Microascaceae
Mycologia 1998; 90:297-302
Abbott SP, Sigler L, Currah RS
Five isolates of Scopulariopsis brevicaulis have been found to form perithecia, in addition to typical conidia. The teleomorph is described as Microascus brevicaulis sp. nov., characterized by small (70 -130 m m diam), black, ostiolate ascocarps which are papillate or very short-necked, and with a peridium of textura angularis. The ascospores are reniform, 5 -6 x 3.5 - 4.5 m m, smooth, and pale orange in mass. Although other species of Microascus produce a Scopulariopsis state in culture, this discovery now definitively links S. brevicaulis, the type species of Scopulariopsis, to the Microascaceae.
Degradation of hydrocarbons in crude oil by the ascomycete Pseudallescheria boydii (Microascaceae)
Canadian Journal of Microbiology 1998; 44: 270-278
April T M, Abbott SP, Foght JM, Currah RS
Four unique strains of Pseudallescheria boydii were isolated from oil-soaked soils in British Columbia and Alberta and compared to strains from cattle dung and from raw sewage. Considerable variability in morphology, colony appearance, colony diameter and temperature tolerance occurred among the strains. They also varied in the sporogenous states produced in culture; all strains had a Scedosporium anamorph and either the Graphium anamorph or cleistothecial teleomorph. Conspecificity of the six isolates was inferred from their morphologies and supported by restriction fragment length polymorphism profiles of the internally transcribed spacer region of rDNA and comparing these to Petriella sordida, a similar taxon in the Microascaceae. Three of the strains isolated from oil-contaminated soil and the strain from sewage were tested for their ability to utilize hydrocarbons by incubation with Prudhoe Bay Crude oil as the sole carbon source. Gas chromatographic analysis of the residual oil revealed that the strains isolated from oil-contaminated soil degraded the linear aliphatics. The strain from sewage, previously shown by others to utilize the volatile n-alkanes (i.e. ethane, propane and butane), did not utilize the liquid saturate compounds. None of the strains was observed to degrade compounds in the aromatic fraction.Pseudallescheria boydii may be an important agent for in-situ bioremediation of saturates in oil-contaminated sites.
Fatal mycotic dermatits in captive brown tree snakes (Boiga irregularis).
J. Zoo Wildlife Medicine 1999; 30 (1):111-118
Nichols DK, Weyant RS, Lamirande EW, Sigler L, Mason RT
Cutaneous fungal infections occurred in four captive brown tree snakes (Boiga irregularis). The ventral scales were most commonly affected and lesions began as areas of erythema and edema with vesicle formation, followed by development of caseous brown plaques. Lesions usually started where ventral scales overlapped and spread rapidly. All snakes died within 14 days after clinical signs were first noted. The deaths of three of the snakes were directly attributable to the cutaneous disease; the other snake died from renal failure and visceral gout most likely induced by gentamicin therapy. Histologically, lesions consisted of epidermal hyperplasia and hyperkeratosis with foci of epidermal necrosis, intra-epidermal vesicle formation, and subacute inflammation of the underlying dermis. These lesions were associated with the presence of bacteria and numerous septate branched fungal hyphae within the epidermis and overlying serocelluar crusts. Hyphae which penetrated through the superficial surface of the epidermis often formed terminal arthroconidia. The same species of fungus was isolated in pure culture from the skin of three snakes, but fungal cultures were not performed on samples from the fourth snake. The fungus has been identified as the Chrysosporium anamorph of Nannizziopsis vriesii based on its formation of solitary dermatophyte-like aleurioconidia and alternate and fission arthroconidia. The source of the fungus in this outbreak was not determined; however, the warm, moist conditions under which the snakes were housed likely predisposed them to opportunistic cutaneous fungal infections.
The genus Uncinocarpus (Onygenaceae) and its synonym Brunneospora: new concepts, combinations and connections to anamorphs in Chrysosporium, and further evidence of its relationship with Coccidioides immitis
Canadian Journal of Botany 1998; 76: 1624-1636
Sigler L, Flis AL, Carmichael JW
The genus Uncinocarpus (Onygenales, Onygenaceae) is emended to include keratinophilic fungi with discrete globose gymnothecial ascomata without differentiated ascomatal hyphae and bearing uncinate, helical or no appendages; oblate, punctate ascospores sometimes with irregular reticulations; bulbous initials, and Malbranchea or Chrysosporium anamorphs. The new combination Uncinocarpus orissi is proposed for Pseudoarachniotus orissi; Gymnoascus arxii is shown to be a synonym. New records show that the fungus has a wide distribution from North America, Europe, Asia and Middle East. The teleomorph is formed under laboratory conditions by mating representative isolates. The anamorph has been described under the names Chrysosporium zonatum and C. gourii . Chrysosporium queenslandicum is morphologically similar. Its teleomorph Apinisia queenslandica is transferred also to the genusUncinocarpus as U. queenslandicus. Brunneospora reticulata, the type species of the genus Brunneospora, is a synonym. Orromyces spiralis appears to be another name applied to this fungus. . Development of helical coils in an isolate of Coccidioides immitis provides further evidence of possible relationship between this dimorphic human pathogen and this group of ascomycetes.
Biosafety and Regulatory Considerations in Handling and Dispatch of Medically Important Fungi in a Culture Collection
Sigler L, Flis A
Sigler & Flis contribution to 1997 ISHAM symposium "Biosafety considerations in handling medically important fungi" For complete paper see, Padhye, A.A., J.E. Bennett, M.R. McGinnis, L. Sigler, A. Flis and I.F. Salkin. 1998. Biosafety considerations in handling medically important fungi. Medical Mycology 36(Suppl 1):258-265.
Note that the views which follow are those of the authors and not of the WFCC or ISHAM.
Culture collections, with expertise in medically important fungi and with appropriate containment facilities for handling such fungi, interact with diagnostic mycology laboratories in several ways. These include: 1) providing taxonomic expertise and professional training; 2) receiving unusual, atypical or rare opportunists for identification and evaluating their potential pathogenic significance; 3) sending reference strains for quality assurance, teaching or research; 4) depositing strains which are the subject of case reports, molecular, epidemiological or other research; 5) conserving biodiversity of medically important fungi; and 6) advising on procedures and regulations concerning safe handling, hazard (risk) levels, requirements for permits, transport, packaging, etc. Regulations governing the safe handling, containment, classification of agents according to risk, packaging and shipping of biological materials and infectious agents, and the prohibitions governing infectious agents deemed as possible biological weapons, are matters of significant concern to culture collections. As regulations become more stringent, often with little influence from the scientific community, they place increasing financial and administrative burdens upon culture collections, and have the potential to significantly impede the exchange of microorganisms for scientific use.
There can be no question that some regulations are necessary for safe handling of microorganisms to help prevent exposure in the workplace and in transit. What is problematic is the lack of harmonization of regulations and differing levels of stringency. Particular problems arise in transport and hazard (risk) level classification. Members of the World Federation of Culture Collections (WFCC) recently agreed that "consideration be paid by regulatory authorities to the essential need for the exchange of microoganisms for scientific and development purposes. To facilitate this process the harmonization of transport regulations, hazard classification and nomenclature is of paramount importance" [1]. Requirements for proper packaging [2,3] have been a major improvement to ensure safe shipping of biological material. However, the international shipment of cultures has become complicated, time consuming and costly, and an impediment to culture exchange. Transport of cultures across national borders is governed by many agencies including International Air Transport Association (IATA) and by domestic regulations which vary between countries [3]. The shipper is responsible for safe transport according to the risk level. Microorganisms (including fungi) are assigned to risk (hazard) groups according to their pathogenic potential. While definitions of risk groups are generally agreed, there is no international consensus on assignment of species. Non-infectious perishable biological substances - Risk group 1 organisms - are not regulated. Infectious substances (IATA) or etiologic agents (USA) are placed in Risk groups 2 or 3 for fungi. These organisms are regulated and must be sent as dangerous goods. The procedures for transport of dangerous goods require trained personnel, engender higher costs, generate a large amount of paperwork and often foster a fear of hazards that may be more perceived than real especially for Risk group 2 organisms.
The WFCC committee on Postal, Quarantine and Safety [4] which is monitoring regulatory changes worldwide is concerned with the tendency for some organisms to be listed in higher level hazard groups, often with little supporting evidence. Moreover, there is a tendency for such lists to form the basis of compilations by interested nations without critical scrutiny and to result in circular reinforcement [4]. Increased stringency in risk levels impacts transport, containment and quarantine requirements. In Canada for example, "Trichophyton species" are classified as Risk Group 2 [5] and thereby encompass the soil keratinophiles T. terrestre and T. ajelloi. This listing may meet the needs of regulators by simplifying bureaucracy but results in the requirement to send relatively benign microbes as dangerous goods, and surely against the priniciple of dangerous goods regulations. Recently the Working Group on Hazardous Fungi (European Confederation of Medical Mycology) recommended reclassification of many species of medically important fungi into biosafety levels according to ecological criteria rather than host factors [6]. Many species were raised to Biosafety level 2 including a number of relatively innocuous species such as the two mentioned above [6]. A concern is that adoption of such a list by regulatory agencies would necessitate the shipment of many additional species as dangerous goods. Moreover the continued placement of some of the dimorphic fungi in level 3 needs critical reexamination. For example, Paracoccidioides brasiliensis is often sterile in the filamentous form and is not considered particularly hazardous or highly infectious by leading workers in paracoccidioidomycosis [7]. Clearly the proposals of this paper [6] should be a basis for further discussion in these critical areas and involve a number of societies and agencies such as ISHAM, WFCC, etc. The exchange of microorganisms is fundamental to research and diagnosis. We need to take a more active role in providing more accurate data and in developing relevant legislation so that this exchange is not impeded.
References
- Anon. World Federation for Culture Collections' Resolutions. WFCC Newsletter December 1996.
- Brown, E.M. & F.P. Simione. ATCC Guide to Packaging and Shipping of Biological Materials. American Type Culture Collection, Rockville, MD, 1994.
- Rohde, C. & D. Claus. 1995. Shipping of Infectious, Non-infectious and Genetically Modified Biological Materials International Regulations. Deutsch Sammlung von Mikrooganismen, Braunschweig, Germany, 1995.
- Smith, D. Committee on postal, quarantine and safety regulations report (1994-1996). World Federation for Culture Collections, Braunschweig, Germany, 1996.
- Kennedy, M.E. Laboratory safety guidelines. Health Canada, Ottawa, ON, 2nd ed. 1996.
- De Hoog, G.S. Risk assessment of fungi reported from humans and animals. Mycoses 1996: 39; 407-417.
- Restrepo-Moreno, A. & D.L. Greer. Paracoccidioidomycosis. Pp 43-64. IN: Occupational Mycoses, A.F. di Salvo, ed., Lea & Febiger, Philadelphia, PA, 1983.
Maxillary sinusitis caused by Schizophyllum commune and experience with treatment.
Journal Medical Veterinary Mycology 1997; 35(5):365-70
Sigler L, Estrada S, Montealegre NA, Jaramillo E, Arango M, De Bedout C, Restrepo A
A case of sinusitis caused by the basidiomycete Schizophyllum commune is reported in a 36-year-old female with a history of allergic rhinitis and dermatitis. The patient presented with sudden nasal obstruction, purulent nasal discharge, headache and general discomfort. Computer tomography revealed extensive opacity of the left maxillary sinus as well as erosion of the nasal wall and maxillary bone. Mycological examinations of nasal discharges and material aspirated during anthrostomy showed hyaline, septate hyphae with rare spicules. Primary isolation yielded a white, woolly mould which demonstrated clamp connections and basidiocarp primordia but these characteristics were lost in subculture. Identification was confirmed by vegetative compatibility studies. The patient was treated with itraconazole to avoid possible postsurgical dissemination. Three months after cessation of therapy, no recurrence of infection had occurred.
Keratomycosis in a Percheron cross horse caused by Cladorrhinum bulbillosum.
Journal Medical Veterinary Mycology 1997; 35(1):53-5
Chopin JB,
Sigler L, Connole MD, O'Boyle DA, Mackay B, Goldstein L
This report describes an infection of a horse's cornea caused by Cladorrhinum bulbillosum. Minor surgery and treatment with antibiotics successfully resolved the infection. The only previous reported case involving this fungus was an Argentinian boy who was infected while working with horses.
Ajellomyces crescens sp. nov., taxonomy of Emmonsia spp., and relatedness with Blastomyces dermatitidis (teleomorph Ajellomyces dermatitidis).
Journal Medical Veterinary Mycology 1996; 34(5):303-14
Sigler L
Adiaspiromycosis is known primarily as a pulmonary infection of small burrowing mammals and rarely of humans, in which the tissue spore form consists of a large, globose, thick-walled, non-proliferating structure called an adiaspore. The causative agents have been placed in
Emmonsia or
Chrysosporium and treated as either two species or varieties.
Emmonsia parva (=
Chrysosporium parvum var. parvum) has been distinguished from
E. crescens (=
C parvum var. crescens) by differences in maximum growth temperature, size of adiaspores, host range and geographical distribution. Phenotypic similarities between
Emmonsia spp. and
Blastomyces dermatitidis and chance observation of
Ajellomyces-type ascomatal hyphae led to the hypothesis that the teleomorph of
Emmonsia spp. could occur in
Ajellomyces. Isolates preliminarily identified as
E. parva or
E. crescens were examined by morphology and physiology and tested for compatibility in mating experiments.
Ajellomyces crescens Sigler sp. nov. is described for the teleomorph of
Emmonsia crescens based on compatibility among 12 of 22 strains, stellate gymnothecial ascomata composed of obtuse diamond-shaped cells, helically coiled appendages and small, globose, muriculate ascospores. The agents of adiaspiromycosis are here treated as species with adiaspore size and morphology and temperature of induction as their major defining features. The species differ also in cycloheximide tolerance and in their abilities to form a teleomorph. With evidence of a connection between
Emmonsia crescens and a teleomorph in
Ajellomyces,
Emmonsia is favoured over
Chrysosporium as the correct name for the agents of adiaspiromycosis. This finding also corroborates earlier suggestions of a close phylogenetic relationship between
Emmonsia spp. and the dimorphic pathogens
Blastomyces dermatitidis and
Histoplasma capsulatum.
Assessment of worker exposure to airborne molds in honeybee overwintering facilities.
American Industrial Hygiene Association Journal 1996; 57(5):484-90
Sigler L, Abbott SP, Gauvreau H
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)/m
3 prior to disturbance by workers and from 2200 to 13,931 CFU/m
3 while workers swept up dead bees. Levels of airborne molds recovered during annual cleaning of beekeeping equipment ranged from 300 to 54,700 CFU/m
3 with an average of 16,083 CFU/m
3. 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.
New records of nail and skin infection due to Onychocola canadensis and description of its teleomorph Arachnomyces nodosetosus sp. nov.
Journal Medical Veterinary Mycology 1994; 32:275-285
Sigler L, Abbott SP, Woodgyer A
Non-dermatophytic fungi are increasingly being recognized as agents of onychomycosis. In 1990, three cases of chronic infection of the great toenail in adult female residents of Canada were attributed to
Onychocola canadensis, a previously unknown hyphomycete. Three additional cases were suspicious but unconfirmed. This report documents seven new records, including six of toenail infection in elderly individuals and one case of glabrous skin infection. Three isolations from New Zealand represent the first report of
O. canadensis outside Canada. Treatment with griseofulvin in one New Zealand hallux infection case was found to improve the appearance of the nail, but specimens were culture positive after 6 months. The development in culture of broad, brown, nodose, thick-walled hyphae suggested an affinity to the ascomycete genus
Arachnomyces. Although mating experiments were attempted on several different media, ascocarps were produced in six mated pairs on sterilized rice grains or rice extract agar after 7-12 months incubation.
Arachnomyces nodosetosus Sigler & Abbott sp. nov. is described and compared with
Arachnomyces minimus Malloch & Cain, also rarely isolated from cutaneous specimens. The genus
Arachnomyces is placed in the
Gymnoascaceae (
Onygenales).
Toenail infection caused by Onychocola canadensis gen. et sp. nov.
Journal Medical Veterinary Mycology 1990; 28(5):405-17
Sigler L, Congly H.
Three cases of great toenail infection are described in which a slow-growing arthroconidial hyphomycete was isolated repeatedly and in pure culture. Direct microscopy revealed hyaline, round to barrel-shaped arthroconidia, hyaline hyphae of varying width, and broad thick-walled brownish hyphae. Three additional isolates were obtained from clinical specimens, for which the results of direct microscopy were unknown or negative. The fungus was resistant to cycloheximide, sensitive to common antifungal drugs by susceptibility tests in vitro and sensitive to benomyl. It was urease positive, hydrolysed casein and tyrosine but not xanthine or hypoxanthine, showed no specific nutritional requirements but grew better on carbohydrate-free media, assimilated 12 carbohydrates and potassium nitrate, and failed to perforate hair. The fungus is described as
Onychocola canadensis Sigler gen. et sp. nov., and it is compared to
Scytalidium lignicola,
Scytalidium hyalinum and the
Scytalidium synanamorph of
Nattrassia mangiferae (
Hendersonula toruloidea).