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Premature osteoarthritis after anterior cruciate ligament reconstruction (ACLR) is common among athletes. Reduced knee contact forces after ACLR likely contribute to the multifactorial etiology of the disease. Whether this reduction is accompanied by compensatory increases in joint contact forces (JCF) at adjacent or contralateral joints is unclear. It is also unclear if compensatory effects depend on the task demands. Thus, we compared hip, knee, and ankle JCF symmetry between individuals with reconstruction and a matched control group during walking and running. Thirty participants (19 females), 2-7 years post-unilateral ACLR (mean = 47.8 months), and 30 controls matched on sex, mass, and activity level were recruited. Limb symmetry indices of peak contact forces and force impulses were calculated for each joint during walking and running, and analyzed using two-factor (group, activity) analysis of variances. Lower ACLR group peak knee JCF (p = 0.009) and knee JCF impulse (p = 0.034) during walking and running were observed. An interaction of group and activity was observed for peak hip JCF, with ACLR participants demonstrating greater involved limb peak hip JCF during running (p = 0.012). Ankle JCF and ground reaction force symmetry indices were not different between groups or across tasks. Decreased knee and increased ipsilateral peak hip JCF during running suggests that proximal adaptations exist at 2-7 years after ACLR, particularly during activities with increased task demand. Clinical significance: Knee and hip JCF asymmetry at 2-7 years after ACLR may underscore a need for clinical strategies and follow-up assessments to identify and target such outcomes., (© 2023 The Authors. Journal of Orthopaedic Research® published by Wiley Periodicals LLC on behalf of Orthopaedic Research Society.)

Introduction: Exophiala spp. are important opportunist pathogens causing subcutaneous or even fatal disseminated infections in otherwise both immunosuppressed and healthy individuals but there are no systematic studies on the isolates of Exophiala species from India., Methods: Twenty-four isolates of Exophiala species were retrieved from the National Culture Collection of Pathogenic Fungi (NCCPF) and identified phenotypically and by molecular methods (ITS region sequencing) followed by antifungal susceptibility testing (AFST) as per CLSI-M38A3 guidelines. A review of the literature of cases from India was performed up to 1 st January 2021 using the Medline and Cochrane database., Results: E. dermatitidis (n = 8), E. jeanselmei (n = 6),  E. spinifera (n = 6),  E. mesophila (n = 1), E. oligosperma (n = 1), E. xenobiotica (n = 1) were identified and the sequencing of ITS, β-tubulin and β-actin revealed a novel species, E. arunalokei sp. nov. (n = 1). The ITS sequence phylogram of E. jeanselmei revealed that the majority (83%) formed a separate cluster close to type A while majority (75%) of E. dermatitidis were type B. The MIC50 (mg/L) of amphotericin, itraconazole, voriconazole, micafungin, caspofungin, anidulafungin, and posaconazole, was 1, 0.25, 0.125, 0.12, 0.125, 0.062, and 0.062, respectively. Sixteen more cases were identified on the literature review and a significant association of E. dermatitidis with history of surgical procedures (p = 0.013), invasive disease (p = 0.032) and of E. mesophila with tuberculosis (p = 0.026) was seen., Conclusion: This, to the best of our knowledge is the first study from India elucidating the molecular and clinical characteristics of Exophiala species and the first Indian report of human infection due to E. xenobiotica and E. arunalokei., Competing Interests: Author BH was employed by the company Hitech Laboratories. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Singh, Rudramurthy, Padhye, Hemashetter, Iyer, Hallur, Sharma, Agnihotri, Gupta, Ghosh and Kaur.)

In this report, we present a case of rhino-orbital mucormycosis in a 57-year-old female with poorly controlled diabetes mellitus. The only mold cultured at 25°C, 37°C, and 40°C from a specimen of the nasal crust was identified phenotypically and independently using nuclear ribosomal DNA sequence data as Thamnostylum lucknowense. To our knowledge, this report presents the first data implicating this mucoraceous fungus as a mycotic agent of human infection.

In this article, we describe a chronic case of rhinofacial mucormycosis caused by Mucor irregularis, formerly known as Rhizomucor variabilis var. variabilis, a rare mycotic agent in humans. The infection caused progressive destruction of the nasal septum and soft and hard palate, leading to collapse of the nose bridge and an ulcerative gaping hole. The mucoralean mold cultured from a nasal biopsy specimen was determined by multilocus DNA sequence data to be conspecific with M. irregularis.

We describe the third fatal case of imported coccidioidomycosis in India in a 31-year-old mechanical engineer originally from Andhra Pradesh, India, who lived in Gwinner, North Dakota. He had traveled to Arizona in summer of 2006, where he drove tractors in a dusty environment at a tractor production facility near Phoenix, Arizona. He was human immunodeficiency virus (HIV) positive. Initially, he was treated in Fargo, North Dakota, in 2006, with liposomal amphotericin B followed by oral fluconazole. Antiretroviral treatment for HIV infection was started. He moved back to India and was admitted to the intensive care unit of St. John's Medical College and Hospital, Bangalore, India. His blood cultures yielded Coccidioides sp. The identity of the isolate was confirmed using the Gen Probe Accuprobe test at the Centers for Disease Control and Prevention, Atlanta, Georgia. In spite of initiation of treatment with antifungal agents (amphotericin B and fluconazole), his condition deteriorated and he expired three days following his admission to the hospital., (Copyright © 2010. Published by Elsevier SAS.)

We report the first two proven cases of cavitary pulmonary zygomycosis caused by Rhizopus homothallicus. The diagnosis in each case was based on histology, culture of the causal agent, and the nucleotide sequence of the D1/D2 region of the 28S ribosomal DNA.

This study examined 215 samples of soil from burrows of rats, other sites in bamboo plantations in different parts of India and Nepal by dilution plating and mouse passage technique for occurrence of Penicillium marneffei and other pathogenic fungi. None of the samples including 25 collected from the burrows of a bamboo rat (Cannomys badius) known to be a carrier of P. marneffei, was positive for the fungus. Among the pathogenic fungi recovered were four isolates of Pseudallescheria boydii (including one from Nepal), two of Trichosporon asteroides, one of Scytalidium hyalinum, 23 isolates of Trichophyton mentagrophytes var. mentagrophytes (including two from Nepal), and two of Microsporum gypseum. Fourteen of the 23 isolates of T. mentagrophytes var. mentagrophytes when tested with the mating types of Arthroderma vanbreuseghemii were found to be of the '+' mating type. The frequent recovery of this dermatophyte from soils of bamboo plantations in several parts of India is remarkable. The study also demonstrates for the first time the occurrence of P. boydii and T. mentagrophytes var. mentagrophytes in Nepalese soil. Among the other fungi recovered were several isolates of species of Aspergillus, Penicillium, Paecilomyces, Fusarium, Chrysosporium, Acremonium, Rhizopus, Mucor, Geotrichum, Trichosporon and Rhodotorula.

A case of mucocutaneous nasal histoplasmosis in an immunocompetent host is described below. A 30-year-old male had a broadened nose with swelling and repeated blockage of nasal passages for the past six months. Diagnosis was made on the basis of histological demonstration of characteristic yeast cells of Histoplasma capsulatum var. capsulatum occurring within histiocytes and extracellularly in stained smears of fine needle aspirates and biopsy from the lesions in ala of the nose and perioral region. The patient showed appreciable regression of lesions after three weeks of itraconazole therapy but was not available for re-assessment.

We describe the first case of white grain pedal eumycetoma caused by Phaeoacremonium krajdenii in a 41-year-old man from Goa, India. Based on histological examination of biopsy tissue showing serpentine granules, a culture of the granules yielding phaeoid fungal colonies, and morphological characteristics and sequence comparison of the partial beta-tubulin gene with the ex-type isolate of P. krajdenii, the causal agent was identified as P. krajdenii.

We describe a case of blastomycosis in a diabetic patient from South India who had visited Milwaukee, Wisconsin, an endemic area for blastomycosis in the USA. After his return to Bangalore, India, the patient developed intermittent fever of moderate to high grade, cough, loss of weight and appetite, and abscesses in the left cubital fossa and thigh regions. Systemic examination at our hospital revealed that he had dullness to percussion over the chest region and decreased breath sounds. Direct examination of Gram-stained smears of the pus from an abscess showed many broad-based budding yeast cells and culture yielded a dimorphic fungus later identified as Blastomyces dermatitidis. Histologic examination of the curettage tissue slides stained with hematoxylin and eosin, periodic acid Schiff's reagent, and Gomori's methenamine silver stain procedures showed many broad-based budding cells characteristic of B. dermatitidis. The patient was successfully treated, initially with amphotericin B, followed by oral itraconazole for a period of 6 months. Blastomycosis cases in India are reviewed and the likely source of infection in this patient is discussed.

Members of the Fusarium solani species complex (FSSC) are increasingly implicated as the causative agents of human mycoses, particularly in the expanding immunocompromised and immunosuppressed patient populations. Best known as ubiquitous plant pathogens and saprotrophs, the FSSC comprises over 45 phylogenetically distinct species distributed among three major clades. To identify which species are associated with human infections, we generated multilocus haplotypes based on four partial gene sequences from 471 isolates. Of these, 278 were from human patients, 21 were from hospital environments, and 172 were from other sources. Phylogenetic trees inferred from an ergosterol biosynthesis gene (erg-3) were highly discordant with those inferred from the three other partial gene sequences; therefore, this partition was analyzed separately. Multilocus analysis showed that isolates from humans were restricted to but spread throughout clade 3 of the FSSC phylogeny, comprising at least 18 phylogenetically distinct species. The majority (74.5%) of the clinical isolates, however, were associated with four major lineages, designated groups 1 to 4. Groups 1 and 2 were strongly supported as phylogenetic species, whereas groups 3 and 4 were not. Although isolates from ocular infections were found in all four groups, they had a significant tendency to belong to group 3 (P < 0.001). Human clinical isolates shared identical multilocus haplotypes with isolates from plants, other animals, and from hospital environments, suggesting potential nosocomiality. The major finding of this study is that FSSC-associated mycoses of humans and other animals have origins in a broad phylogenetic spectrum, indicating widespread ability to cause infection in this diverse species complex.

We describe a case of concurrent double infection with phaeohyphomycosis and lymphocutaneous nocardiosis in an immunocompromised host. The episode occurred almost simultaneously with an incident of insect bites. Exophiala spinifera was isolated from lesions on the left arm and Nocardia asteroides was isolated from lesions on the right hand. Clinical resolution occurred within 5 months after initiation of antimicrobial treatment with oral itraconazole and minocycline. We believe that this case represents a novel episode of previously unreported concurrent infections with phaeohyphomycosis and lymphocutaneous nocardiosis in human beings.

Background: During December 2000-July 2001, black sediment was noted in saline-filled silicone breast implants of women who had undergone revision surgery at facility A. Curvularia fungus was isolated from implant saline., Methods: To identify risk factors for contamination with Curvularia species, we performed case-control, retrospective cohort, and laboratory studies and conducted procedural reviews. A case patient was defined as any woman who underwent revision surgery at facility A between January 2000 and June 2001 and had black sediment in her implants., Results: Five patients met the case definition. Contamination was associated with having had surgery performed in operating room (OR) 2 (4/88 vs. 1/140; P=.07) and a longer duration of surgery (P<.001). A longer duration spent in the OR was an additional risk factor (P=.005). Curvularia fungus was isolated from the sterile supply room, where saline bottles had been stored under a water-damaged ceiling, and from the corridor outside OR 2; it was also found more commonly from facility A personnel than from non-facility A personnel (12/34 vs. 4/60; P<.001). Saline was warmed in a cabinet opposite OR 2, which was maintained at negative pressure differentials, then was poured into bowls open to the OR 2 environment before injection into implants., Conclusion: Surgeons should always use closed systems to inflate breast implants. Surgery center infection control measures must include moisture control and balanced ventilation systems.

To date, three species of Phaeoacremonium have been associated with phaeohyphomycosis. These are P. parasiticum (formerly Phialophora parasitica), P. inflatipes, and P. rubrigenum. Numerous unknown isolates resembling Phaeoacremonium spp. have in recent years been isolated from human patients as well as from woody plants that appear to be the main environmental source of these fungi. Nine new Phaeoacremonium species, of which six were obtained as etiologic agents of human opportunistic infection, are reported. They can be identified based on their cultural and morphological characters, and the identifications are strongly supported in phylogenetic analyses of partial sequences of the actin, beta-tubulin, and calmodulin genes. A multiple-entry electronic key based on morphological, cultural, and beta-tubulin sequence data was developed to facilitate routine species identification. Reexamination of all isolates of P. inflatipes associated with human disease showed them to be misidentified and to belong to the new taxa described here.

Fungal valvular endocarditis is an unusual cause of endocarditis, yet very important because of its historically poor prognosis. We report two fatal cases of fungal valvular endocarditis following cardiovascular surgery, presenting as femoral artery embolism. Aspergillus terreus and A. flavus were the causative agents of endocarditis in the two patients. Diagnosis was established very early by culture of the emboli and was confirmed later by isolation of the same Aspergillus species from the resected valve tissue.

This report describes severe generalized granulomatous lymphadenitis and splenitis in a 5-year-old, spayed female, Rottweiler dog with anorexia and diarrhea. There was replacement or effacement of much of the parenchyma of the lymph nodes and spleen by sheets of macrophages, multinucleated giant cells, and myriad nonpigmented fungal organisms, most of which appeared to be intracellular. These organisms were very pleomorphic, including large chlamydospore-like cells, small round yeast-like cells, and septate hyphae. A fungus identified as Monocillium indicum was isolated from lymph node tissue. To the authors' knowledge, this is the first report of infection with Monocillium in either humans or other animals.

A 41-year-old man with a past medical history of diabetes mellitus type II, AIDS (diagnosed 2 years earlier; CD4 count < 10), peripheral neuropathy, and pulmonary tuberculosis of 2 years duration was admitted to the hospital with abnormal liver function tests. There was a chronic hepatitis/cholestasis that had worsened while the patient was undergoing directly observed tuberculosis therapy. On admission, the patient complained of a painful swelling on his right arm. In the posterior aspect of the arm, there was a 3-4-cm subcutaneous mass that was fluctuant, mobile and tender. Incision of the mass released yellowish pus mixed with blood. Direct examination of the pus in KOH mounts and Gram-stained smears revealed subhyaline, septate, branched hyphae. When the pus was cultured on Sabouraud dextrose agar containing chloramphenicol, several velvety, olivaceous grey colonies grew after 7 days at 25 degrees C. When grown on oatmeal agar, the fungus produced subglobose, rostrate pycnidia with phialidic conidiogenous cells, and 1-celled cylindrical conidia. It was identified as Pleurophomopsis lignicola Petrak. This report describes the third known case of subcutaneous infection caused by P. lignicola in an immunocompromized patient.

The second case of phaeohyphomycosis caused by Veronaea botryosa in China, in a 12-year-old boy from Jiangsu Province, is presented. Based on direct examination of the scrapings from crusted lesions; histologic examination of the biopsy tissue showing septate, phaeoid hyphal elements; and the culture exhibiting sympodial, conidiogenous cells producing predominantly two-celled, cylindric conidia, the etiologic agent was identified as V. botryosa.

A 45-year-old man from Nepal with a 13-year history of polycystic kidney disease was diagnosed as suffering from chronic renal failure with end-stage renal disease. After receiving empirical antituberculosis treatment, he was treated with broad-spectrum antibiotics. A left nephrectomy was performed, and after 4 months, he received a kidney transplant. The left kidney was grossly enlarged, with multiple cystic spaces filled with blackish material. Histologic examination of the excised left kidney tissue stained with hematoxylin and eosin and Gomori's methenamine silver stains showed numerous hyaline, septate, fungal hyphae of various lengths, many broken into rectangular arthroconidia in the cystic spaces. Culture of the kidney tissue yielded white, glabrous, yeast-like colonies. Based on its micromorphology, growth at 42 degrees C, and ribosomal DNA (rDNA) sequence analysis, and also sequence analysis of the internal-transcribed-spacer and D1/D2 rDNA regions, the yeast was identified as Trichosporon loubieri. Postsurgically, the patient was treated with amphotericin B and oral itraconazole, followed by maintenance therapy with fluconazole. He remained afebrile and asymptomatic. At the final follow-up, all parameters were found normal and the patient was doing well, with normal renal function reports. This paper presents the first known case of human infection caused by T. loubieri.

We describe a fatal case of imported coccidioidomycosis in India in a 22-year-old male who worked in Tucson, Arizona, approximately four years prior to his illness. The diagnosis was based on the presence of characteristic spherules with endospores in biopsy tissue of lymph nodes, bone and pus from a chronic discharging sinus in the left gluteal region and isolation of Coccidioides immitis in culture. C. immitis is one of the most infectious and virulent fungal pathogens and poses a serious occupational hazard for laboratory personnel, especially in areas where the disease is not endemic. To reduce the role of laboratory-acquired infection, all procedures that involve manipulation of cultures of C. immitis should, whenever possible, be conducted in a biological safety cabinet.

We describe the first case of cryptococcosis caused by Cryptococcus neoformans var. gattii in a male Atlantic bottlenose dolphin (Tursiops truncatus). The dolphin showed clinical signs of tachypnea, transient dyspnea, and mild tachycardia and developed multiple hyperechoic nodules, parenchymal consolidation, and thickening of pleura. A diagnosis of bronchopneumonia with pleuritis was made. Itraconazole therapy was implemented for 120 days, and trough levels in serum were within or above the suggested therapeutic range. Titers of cryptococcal antigen in serum increased eightfold during therapy, and the case had a fatal outcome. Necropsy examination findings included enlarged pulmonary lymph nodes and extensive coalescing granulomatous lesions throughout both lungs. Histologic examination revealed numerous, spherical to ellipsoidal, mucicarmine-positive, 3- to 14-microm, encapsulated, budding cells consistent with C. neoformans. Culture of the lung tissue yielded colonies of C. neoformans. The isolate was urease positive and nitrate negative and exhibited phenoloxidase activity. It was positive on canavanine-glycine-bromothymol blue agar. When tested by the Iatron serodiagnostic reagent kit (Iatron Laboratories, Inc.), it was shown to belong to serotype B.

An outbreak of invasive aspergillosis occurred in a community hospital in temporal association with construction activity. Epidemiological investigation showed that patients who are at highest risk comprise a small group and are readily identifiable. Clinicians should strive to protect these patients, following guidelines published by the Centers for Disease Control and Prevention.

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.

We describe the first human case of lobomycosis caused by Lacazia loboi in a 42-year-old white male resident of Georgia. The patient had traveled to Venezuela 7 years earlier, where he had planned to rappel down Angel Falls in Canaima. Although he never actually rappelled the falls, he did walk under the falls at least three times, exposing himself to the high water pressures of the falls. He noticed a small pustule with surrounding erythema developing on the skin of his right chest wall. The lesion gradually increased in size and had an appearance of a keloid. For cosmetic reasons, the patient sought medical treatment to remove the lesion. After an uncomplicated excision of the lesion, the patient recovered completely. The excised tissue was fixed in formalin for pathologic examination. Tissue sections stained by hematoxylin and eosin, periodic acid-Schiff stain, and Gomori methenamine silver stain procedures showed numerous histiocytes, multinucleated giant cells, and numerous globose or subglobose, lemon-shaped cells producing multiple blastoconidia connected by narrow tube-like connectors and catenate chains of various lengths characteristic of L. loboi.

Candida dubliniensis is a newly described species that is closely related phylogenetically to Candida albicans and that is commonly associated with oral candidiasis in human immunodeficiency virus-positive patients. Several recent studies have attempted to elucidate phenotypic and genotypic characteristics of use in separating the two species. However, results obtained with simple phenotypic tests were too variable and tests that provided more definitive data were too complex for routine use in the clinical laboratory setting. The objective of this study was to determine if reproducible identification of C. dubliniensis could be obtained with commercial identification kits. The substrate reactivity profiles of 80 C. dubliniensis isolates were obtained by using the API 20C AUX, ID 32 C, RapID Yeast Plus, VITEK YBC, and VITEK 2 ID-YST systems. The percentages of C. dubliniensis isolates capable of assimilating or hydrolyzing each substrate were compared with the percentages from the C. albicans profiles in each kit's database, and the results were expressed as percent C. dubliniensis and percent C. albicans. Any substrate that showed >50% difference in reactivity was considered useful in differentiating the species. In addition, assimilation of methyl-alpha-D-glucoside (MDG), D-trehalose (TRE), and D-xylose (XYL) by the same isolates was investigated by the traditional procedure of Wickerham and Burton (L. J. Wickerham and K. A. Burton, J. Bacteriol. 56:363-371, 1948). At 48 h (the time recommended by the manufacturer for its new database), we found that the assimilation of four carbohydrates in the API 20C AUX system could be used to distinguish the species, i.e., glycerol (GLY; 88 and 14%), XYL (0 and 88%), MDG (0 and 85%), and TRE (15 and 97%). Similarly, results with the ID 32 C system at 48 h showed that XYL (0 and 98%), MDG (0 and 98%), lactate (LAT; 0 and 96%), and TRE (30 and 96%) could be used to separate the two species. Phosphatase (PHS; 9 and 76%) and alpha-D-glucosidase (23 and 94%) proved to be the most useful for separation of the species in the RapID Yeast Plus system. While at 24 h the profiles obtained with the VITEK YBC system showed that MDG (10 and 95%), XYL (0 and 95%), and GLY (26 and 80%) could be used to separate the two species, at 48 h only XYL (6 and 95%) could be used to separate the two species. The most useful substrates in the VITEK 2 ID-YST system were TRE (1 and 89%), MDG (1 and 99%), LAT (4 and 98%), and PHS (83 and 1%). While the latter kit was not yet commercially available at the time of the study, it would appear to be the most valuable for the identification of C. dubliniensis. Although assimilation of MDG, TRE, and XYL proved to be the most useful for species differentiation by the majority of commercial systems, the results with these carbohydrates by the Wickerham and Burton procedure were essentially the same for both species, albeit following protracted incubation. Thus, it is the rapidity of the assimilation achieved with the commercial systems that allows the differentiation of C. dubliniensis from C. albicans.

We describe four cases of disseminated infection caused by endemic Penicillium marneffei in human immunodeficiency virus (HIV)-infected patients from the Manipur state of India. The most common clinical features observed were fever, anorexia, weight loss, hepatosplenomegaly, and, more importantly, skin lesions resembling molluscum contagiosum. The diagnosis in each of the four cases was achieved by direct examination of smears, observance of intracellular yeast-like cells multiplying by fission in biopsied tissue from skin lesions, and isolation of the dimorphic P. marneffei in pure culture in each case. In one case, fluorescent antibody studies allowed specific diagnosis. This report documents a new area in which P. marneffei is endemic, located in eastern India, and describes the first occurrence in India of P. marneffei in HIV-infected patients as well as the extension of the areas of P. marneffei endemicity westward to the northeastern state of Manipur.

We describe a case of coccidioidomycosis in which several unusual morphologic forms of Coccidioides immitis occurred in biopsy tissue from the right lower lung of a patient. To our knowledge, this is the first case where so many diverse morphologic forms were manifested in a single patient in the absence of typical endosporulating spherules. Immature spherules demonstrating segmentation mimicked morula forms of Prototheca spp. Certain elements resembled budding cells of Blastomyces dermatitidis. These consisted of juxtaposed immature spherules without endospores, a germinating endospore, or thick-walled hyphal cells. Branched, septate hyphae and moniliform hyphae consisting of chains of thick-walled arthroconidia or immature spherules were also present. Complement fixation and immunodiffusion tests performed on the patient's serum were negative for C. immitis, B. dermatitidis, and Histoplasma capsulatum antibodies. Fluorescent-antibody studies were carried out with a specific C. immitis conjugate. All of the diverse fungal tissue elements stained positive with a moderate to strong (2 to 3+) intensity.

Phaeoacremonium inflatipes, one of three species previously classified as strains of Phialophora parasitica, was identified as the causal agent of a subcutaneous infection of the left foot of an 83-year-old woman from South Carolina. The patient had a granulomatous growth over the anteromedial aspect of her left foot. It was surgically excised, which led to complete healing without complications. Tissue sections of the excised mass stained with hematoxylin and eosin and Gomori's methenamine silver strains showed many septate hyphal elements of various lengths, some exhibiting brownish pigment in the cell walls of the hyphae. Portions of the tissue, when cultured, yielded many colonies which were initially glabrous, off white becoming velvety, greyish brown on aging. Microscopically, their hyphae were septate, branched, and phaeoid and bore lateral and terminal, erect, septate conidiophores. The conidiogenous cells (phialides) were terminal or lateral, mostly monophialidic, subcylindrical to spinelike in shape, and constricted at their bases and bore funnel-shaped, inconspicuous collarettes at their tips. The conidia were subhyaline, oblong, and ellipsoid to allantoid.

We developed a method for the identification of Aspergillus fumigatus fungal isolates by using random amplified polymorphic DNA (RAPD) PCR (RAPD-PCR) cloning and the TaqMan LS50B fluorogenic detection system (Perkin-Elmer Corp., Applied Biosystems, Foster City, Calif.). DNA from seven clinically important Aspergillus species was screened by RAPD-PCR to identify section- or species-specific amplicons. With the OPZ19 RAPD primer a 1,264-bp product was amplified from all A. fumigatus strains initially examined but not from other species. A partial DNA sequence of this product was used to design a specific primer pair, which generated a single 864-bp fragment with DNA from 90 of 100 A. fumigatus isolates when a "touchdown" (65-->55 degrees C) annealing protocol was used. The TaqMan system, a fluorogenic assay which uses the 5'-->3' endonuclease activity of Taq DNA polymerase, detected this 864-bp product with DNA from 89 of these 90 A. fumigatus strains; 1 DNA sample generated an indeterminate result. With DNA from three morphologically typical A. fumigatus isolates, six white ("albino") A. fumigatus isolates, and five of six Neosartorya species (non-A. fumigatus members of the section Fumigati), the 864-bp product was amplified differentially at an annealing temperature of 56 degrees C but not with the touchdown annealing format. No amplicon was detected with DNA from 56 isolates of heterologous Aspergillus, Penicillium, and Paecilomyces species or from Neosartorya fennelliae; TaqMan assay results were either negative (51 isolates) or indeterminate (5 isolates) for all isolates. This RAPD-PCR and TaqMan assay offers promise as a nucleic acid-based system that can be used for the identification of filamentous fungal isolates and that requires no postamplification sample manipulations.

We report the first well documented case of subcutaneous phaeohyphomycotic infection caused by Phialophora verrucosa in India. Examination of the biopsied tissue from an ulcerating lesion on the shin of the left leg of a 45-year-old woman from Bombay, India, showed numerous dematiaceous, septate, branching hyphal elements and thick-walled cells characteristic of phaeohyphomycosis. Cultures of the scrapings from the lesion and of the biopsied tissue yielded a pigmented fungus that was identified as P. verrucosa. Initial treatment with fluconazole followed by oral itraconazole for 30 days and local application of a copper sulphate solution resulted in complete resolution of the lesion. Treatment with itraconazole was continued for an additional 3 months after complete healing. No new lesions developed and the patient did not show jaundice, hepatosplenomegaly or any other signs of toxicity.

Mitochondrial DNA (mtDNA) types based on restriction fragment length polymorphism (RFLP) patterns with HaeIII were investigated in clinical isolates of Sporothrix schenckii in North and South America. In addition to 14 mtDNA types (Types 1-14) so far reported, six new mtDNA types, Types 15-20 were found in this study. Type 3 was divided into two subtypes, Subtype 3A and Subtype 3B based on RFLP with Msp1. Type 14 was also divided into three subtypes, Subtype 14A, Subtype 14B and Subtype 14C based on RFLP with Hha1. Nineteen isolates in the United States consisted of 1 isolate of Type 1, 12 of Type 2, 2 of Type 4, 3 of Type 14 (1 of Subtype 14B and 2 of Subtype 14C) and 1 of Type 15. Twenty nine isolates in Venezuela consisted of 13 of Type 3 (Subtype 3B), 6 of Type 4, 1 of Type 18, 3 of Type 19 and 6 of Type 20. Thirteen isolates in Argentina consisted of 2 of Type 3 (Subtype 3A), 4 of Type 4, 4 of Type 16 and 3 of Type 17. One isolate in Brazil was Type 3 (Subtype 3A). Based on the phylogeny of 20 mtDNA types (Types 1-20) constructed by estimating sequence divergences of mtDNA, mtDNA types were clustered into two groups: Group A (Types 1-3, Type 11 and Types 14-19) and Group B (Types 4-10, Types 12-13 and Type 20). These results suggest that S. schenckii isolates in North and South America mainly belong to Group A.

Over 500,000 workers in the USA alone are employed in laboratories that range from small physician offices to large clinical laboratories handling microbes for comprehensive research and/or diagnostic work. These workers are exposed to a variety of potential occupational health risks such as exposure to infectious clinical materials, environmental specimens, cultures, complex and inflammable chemicals, radiation, and electrical and mechanical hazards. As members of the International Society for Human and Animal Mycology, we have no policy statement on biosafety standards for handling medically important fungi. The intent of the symposium is to cover some of the important aspects of biosafety; (1) standards in handling dimorphic fungal pathogens; (2) the principles and criteria of biosafety levels and classification of known medically important fungi, aerobic actinomycetes, environmental fungi according to their biosafety levels; (3) medically important fungal waste and its safe disposal; and (4) biosafety and regulatory considerations in handling and mailing medically important fungi in a culture collection.

Cryptococcus neoformans var. gattii has an ecological association with five Eucalyptus species: E. blakelyi, E. camaldulensis, E. gomphocephala, E. rudis, and E. tereticornis. After human infections due to C. neoformans var. gattii were diagnosed in the states of Punjab, Himachal Pradesh, and Karnataka, India, a study was undertaken to investigate the association of C. neoformans var. gattii with Indian eucalypts, especially in the state of Punjab. A total of 696 specimens collected from E. camaldulensis, E. citriodora and E. tereticornis (hybrid) trees were examined for the presence of C. neoformans var. gattii. Flowers from two trees of E. camaldulensis in the Chak Sarkar forest and one from the village of Periana near the Ferozepur area yielded five isolates of C. neoformans var. gattii. The origin of the trees could be traced to Australia, thus providing evidence that the distribution of E. camaldulensis correlated with the distribution of human cryptococcosis cases caused by C. neoformans var. gattii in northern India.

A fatal case of craniofacial zygomycosis caused by Apophysomyces elegans in a 52-year-old man was diagnosed by the presence of broad aseptate, branched hyaline hyphae in tissue from paranasal sinuses and surrounding areas, and isolation of the fungus from the same tissue. The patient suffered from idiopathic myelofibrosis as underlying disease, he was thrombocytopenic and was mildly hyperglycaemic. The infection represents the second case of craniofacial zygomycosis due to A. elegans.

A new medium, Dermatophyte Identification Medium (DIM) (trade mark pending), was specifically developed to eliminate problems of false-positive results associated with commercially marketed media, such as dermatophyte test medium (DTM). Previous investigations had demonstrated that DTM only partially suppressed growth of nondermatophytes and that several of these nondermatophytic fungi that were morphologically similar to dermatophytes caused false-positive results. Presumptive identification of an unknown isolate as a dermatophyte required only the transfer of a portion of the suspected colony recovered from the specimen to DIM. Positive results, evidenced by a change in the color of the medium, were observed within 24 to 48 h. In studies of over 500 isolates of dermatophytes and common nondermatophyte molds, as well as close to 600 yeast isolates, false-positive results were always associated with bacterial contamination of the mold isolates while false negatives were only observed with occasional isolates of Trichophyton verrucosum. DIM culture was an inexpensive, rapid, and accurate method for the presumptive identification of dermatophytes in the clinical mycology laboratory.

An immunocompetent 59-year-old man developed sinusitis over a 6- to 8-month period after cutting down a rotted maple tree (Acer sp.). A polypoid obstruction with a bloody drainage was evident in his right nasal cavity. A computed tomographic scan showed an opacification of the maxillary sinus. Surgery was performed to remove a fungus ball that had extended into the patient's medial sinus cavity. Sections of the sinonasal mucosa revealed marked acute and chronic sinusitis with inflammation, congestion, and hemorrhage. Sections from the pasty brown to black debrided material revealed a fungus ball consisting of an extensive network of brown-pigmented, septate, profusely branched hyphae. When grown on oat agar, the phaeoid fungus produced pycnidia and was identified as Pleurophomopsis lignicola. The genus Pleurophomopsis includes seven species, which are all known from plant material. This report documents for the first time a coelomycetous fungus, P. lignicola, causing sinusitis in an immunocompetent patient.

We report three cases of primary cutaneous zygomycosis due to either Saksenaea vasiformis (two patients) or Apophysomyces elegans (one patient). Extensive surgical debridement helped two patients recover from their infections. The underlying disease in the one patient who died was diabetes mellitus. We reviewed the literature on primary cutaneous zygomycosis and found that S. vasiformis and A. elegans were the etiologic agents in 16 and 13 earlier cases, respectively.

We studied an aberrant culture of Histoplasma capsulatum var. capsulatum isolated from synovial fluid collected from the right elbow of a patient from Kansas. Colonies on Sabouraud glucose agar and other routine mycological media were glabrous to soft, moist, heaped, deeply folded or convoluted, and orange-brown with a white, irregular margin. Microscopically, hyphae were hyaline, septate, and branched and remained totally devoid of conidiation over a period of 2 years on all mycological media. Conversion to the yeast form was achieved on Pine's medium at 37 degrees C. Colonies at early stages of growth were smooth, moist, pasty, shiny, and orange-brown but soon became wrinkled and slightly raised and produced oval, thin-walled cells measuring 2 to 3 by to 4.5 microns which multiplied by polar budding. The identity of the isolate was further confirmed by utilizing the Accuprobe DNA and the exoantigen test for H. capsulatum var. capsulatum.

Exoantigenic extracts of 15 isolates belonging to hyalohyphomycosis-causing Beauveria bassiana (1), and Engyodontium album (1), as well as other species of the genus Beauveria (one isolate each of B. brogniartii, B. densa, B. stephanoderis, B. velata, B. vermiconia and six isolates of unknown Beauveria species) were studied. Aqueous-merthiolated extracts derived from 10-day-old Sabouraud's dextrose agar slant cultures (25 degrees C) were concentrated (25X), and reacted against rabbit anti-B. bassiana serum in the presence of partially purified homologous antigen (20X) prepared from 5-week-old shaken cultures (30 degrees C), using a microimmunodiffusion procedure. Beauveria bassiana reference antigen and antiserum reacted to produce four bands of identity. With the exception of E. album, which was negative, extracts of the isolates of B. brogniartii, B. densa, B. stephanoderis, B. velata, B. vermiconia and the unknown Beauveria species all produced 2-4 lines of identity against the homologous anti-B. bassiana serum. These results suggested that all the species of the genus Beauveria tested were antigenically related to B. bassiana. Engyodontium album demonstrated antigenic distinctness, however, from B. bassiana and thus supported the validity of this taxon.