A case of sporotrichosis caused by different Sporothrix brasiliensis strains: mycological, molecular, and virulence analyses.

BACKGROUND: Sporotrichosis is a subcutaneous mycosis caused by dimorphic pathogenic fungi belonging to the Sporothrix genus. Pathogenic Sporothrix species typically produce melanin, which is known to be a virulence factor. OBJECTIVES: The aim of this study was to perform phenotypic, genotypic, and virulence analyses of two distinct Sporothrix brasiliensis strains isolated from the same lesion on a patient from Rio de Janeiro. METHODS AND FINDINGS: Genotypic analyses by partial sequencing of the calmodulin, ß-tubulin, and chitin synthase genes, as well as polymerase chain reaction (PCR)-fingerprinting by T3B, M13, and GACA, showed that the isolates were very similar but not identical. Both isolates had similar phenotypic characteristics and effectively produced melanin in their yeast forms, accounting for their ability of causing disease in a murine sporotrichosis model. Remarkably, isolate B was albino in its environmental form but caused more severe disease than the pigmented A isolate. CONCLUSIONS: These findings indicate that the patient was infected by two genetically and biologically distinct S. brasiliensis that vary in their production of melanin in their environmental forms. The results underscore the importance of characterizing phenotypically different isolates found in the same clinical specimen or patient.

Clinical features, fungal load, coinfections, histological skin changes, and itraconazole treatment response of cats with sporotrichosis caused by Sporothrix brasiliensis.

Zoonotic sporotrichosis caused by the fungus Sporothrix brasiliensis is usually severe in cats. This study investigated the associations between clinical features, fungal load, coinfections, histological skin changes, and response to itraconazole in cats with sporotrichosis caused by S. brasiliensis. Fifty-two cats with skin lesions and a definitive diagnosis of sporotrichosis were treated with itraconazole for a maximum period of 36 weeks. The animals were submitted to clinical examination and two subsequent collections of samples from the same skin lesion for fungal diagnosis and histopathology, as well as serology for feline immunodeficiency (FIV) and leukaemia (FeLV) viruses. Thirty-seven (71%) cats were clinically cured. Nasal mucosa lesions and respiratory signs were associated with treatment failure. Cats coinfected with FIV/FeLV (n = 12) had a lower neutrophil count in the lesion. A high fungal load in skin lesions was linked to young age and treatment failure, as well as to a longer time of wound healing, poorly formed granulomas and fewer neutrophils, macrophages and lymphocytes in these lesions. These results indicate that itraconazole is effective, but nasal mucosal involvement, respiratory signs and high fungal loads in skin lesions are predictors of treatment failure that will assist in the development of better treatment protocols for cats.

Experimental Hyalohyphomycosis by Purpureocillium lilacinum: Outcome of the Infection in C57BL/6 Murine Models.

Purpureocillium lilacinum is a filamentous, hyaline fungus considered an emerging pathogen in humans. The aim of our study was to evaluate the outcome of hyalohyphomycosis in C57BL/6 murine models inoculated with two clinical P. lilacinum isolates (S1 and S2). Each isolate was inoculated in mice randomly distributed in immunocompetent (CPT) and immunosuppressed (SPS) groups. Mice were evaluated at day 7, 21, and 45 after inoculation for histopathological analysis, recovery of fungal cells, and immunological studies. Histological analysis showed scarce conidia-like structures in lung tissue from CPT mice and a lot of fungal cells in SPS mice inoculated with S2 compared to mice inoculated with S1. The maximum recovery of fungal cells was seen in CPT mice inoculated with both isolates at day 7, but with mean significantly higher in those inoculated with S2 isolate. Phenotypical characterization of T cells showed TCD8+ lymphocytes predominance over TCD4+ in immunosuppressed mice infected and control groups. We also observed higher percentages of the central and effector memory/effector phenotype in CPT mice infected with S2 strain, especially in TCD8+ in the initial period of infection. Regulatory T cells showed higher percentages in immunosuppressed, predominantly after the acute phase. Our results showed that the P. lilacinum is a fungus capable to cause damages in competent and immunosuppressed experimental hosts. Furthermore, S2 isolate seems to cause more damage to the experimental host and it was possible to identify different cellular subsets involved in the mice immune response.

Reflexões sobre o ensino online de Biossegurança à luz da Teoria da Aprendizagem Significativa / Reflections on the online learning of Biosafety based in the meaningful learning

Organizar eventos educacionais, com foco na aprendizagem, exige articulação entre planejamento, processo de ensino e avaliação. O presente manuscrito objetiva avaliar a estruturação pedagógica e os resultados no QBA/Online ­ curso de Biossegurança a distância, inserido no Moodle, oferecido aos ingressos do Instituto Oswaldo Cruz ­ à luz da Teoria da Aprendizagem Significativa (TAS), para identificar oportunidades de melhorias. Na pesquisa qualitativa, foram considerados os dados de 2760 participantes, de 2008 a 2014. A média de participação anual foi de 460, com predominância de alunos (40%). No planejamento, não foi utilizada uma fundamentação teórica-metodológica, mas considerados os aspectos contextuais do ensino e a natureza do conhecimento a ser ensinado. O conteúdo está em 12 textos. O maior índice de acertos nos testes foi de 18 questões, no único instrumento de avaliação da aprendizagem, com 20 perguntas, centradas nos temas dos textos. O curso foi bem avaliado pelos alunos. A flexibilidade do tempo/local está dentre os pontos positivos e a quantidade/tamanho dos textos, nos negativos. O diagnóstico ratificou a relevância de um curso online; necessidade de reorganização do material instrucional; e de oferta de estratégias de ensino para estimular o interesse dos alunos

Organizing educational events, focusing on learning, requires disposition between planning, teaching process and evaluation. This manuscrip tintent to evaluate pedagogical format and results in QBA/Online ­ distance learning course on Biosafety, accessed in Moodle platform, off ered to graduates from Oswaldo Cruz Institute ­ based in the Meaningful Learning Theory as to enlighten opportunities for improvement. A qualitative inquiry was performed, with 2760 participants and registered in the system, from 2008 to 2014. The average annual contribution is 460, with a predominance of students (40%). With regard toplanning, theoretical-methodological were not considered, only contextual aspects of education and the concepts of knowledge to be taught/instructed. Twelve texts were assigned. Thehighest scores rates in tests reached 18 questions, the only instrument for assessing learning, whichhas 20 questions, focusing on themes of current texts. The course was well rated by those who completed the evaluation form. Participants highlighted as positive aspects; flexibility as to time/place and as negative the large number of texts and its sizes. The analysis confirmed the benefits of online tool; the need to rearrange institutional material; and providing teaching strategies to encourage students' interest

Reflexões sobre o Ensino online de Biossegurança à luz da Teoria da Aprendizagem Significativa / Reflections on the online learning of Biosafety based in the meaningful learning

Organizar eventos educacionais, com foco na aprendizagem, exige articulação entre planejamento, processo de ensino e avaliação. O presente manuscrito objetiva avaliar a estruturação pedagógica e os resultados no QBA/Online – curso de Biossegurança a distância, inserido no Moodle, oferecido aos ingressos do Instituto Oswaldo Cruz – à luz da Teoria da Aprendizagem Significativa (TAS), para identificar oportunidades de melhorias. Na pesquisa qualitativa, foram considerados os dados de 2760 participantes, de 2008 a 2014. A média de participação anual foi de 460, com predominância de alunos (40%). No planejamento, não foi utilizada uma fundamentação teórica-metodológica, mas considerados os aspectos contextuais do ensino e a natureza do conhecimento a ser ensinado. O conteúdo está em 12 textos. O maior índice de acertos nos testes foi de 18 questões, no único instrumento de avaliação da aprendizagem, com 20 perguntas, centradas nos temas dos textos. O curso foi bem avaliado pelos alunos. A flexibilidade do tempo/local está dentre os pontos positivos e a quantidade/tamanho dos textos, nos negativos. O diagnóstico ratificou a relevância de um curso online; necessidade de reorganização do material instrucional; e de oferta de estratégias de ensino para estimular o interesse dos alunos (AU)

Organizing educational events, focusing on learning, requires disposition between planning, teaching process and evaluation. This manuscrip tintent to evaluate pedagogical format and results in QBA/Online – distance learning course on Biosafety, accessed in Moodle platform, off ered to graduates from Oswaldo Cruz Institute – based in the Meaningful Learning Theory as to enlighten opportunities for improvement. A qualitative inquiry was performed, with 2760 participants and registered in the system, from 2008 to 2014. The average annual contribution is 460, with a predominance of students (40%). With regard toplanning, theoretical-methodological were not considered, only contextual aspects of education and the concepts of knowledge to be taught/instructed. Twelve texts were assigned. Thehighest scores rates in tests reached 18 questions, the only instrument for assessing learning, whichhas 20 questions, focusing on themes of current texts. The course was well rated by those who completed the evaluation form. Participants highlighted as positive aspects; flexibility as to time/place and as negative the large number of texts and its sizes. The analysis confirmed the benefits of online tool; the need to rearrange institutional material; and providing teaching strategies to encourage students’ interest (AU)

Feline histoplasmosis: a case of supraorbital involvement / Histoplasmose felina: relato de caso com envolvimento supraorbital

Histoplasmosis is a systemic fungal infection caused by the dimorphic fungus Histoplasma capsulatum that infects humans and a wide variety of mammalian species, including cats. In Brazil only four cases of this disease were reported in domestic cats. The present study reports a case of an indoor cat, rescued from street three years ago, that developed histoplasmosis with supraorbital involvement. The cat presented sneezing, dyspnea and increased respiratory sounds and the diagnosis suspected was allergic rhinitis. Prednisolone and amoxicillin were prescribed. The patient returned to clinic after clinical signs worsened with a swelling of the right supraorbital region, unilateral left side conjunctivitis and enlarged submandibular lymph nodes. Cytology and culture revealed Histoplasma capsulatum. Initially, itraconazole (10 mg/Kg SID orally) was prescribed and due to the worsening of clinical signs, was combined amphotericin B (0.5 mg/kg SC) for two weeks. The therapy with amphotericin was discontinued and itraconazole continued to be administered for six months. The clinical signs were resolved and no recrudescence was reported. It is important that veterinary practitioners to be aware of the clinical features of feline histoplasmosis and consider the possibility of a subclinical infection.

Histoplasmose é uma infecção sistêmica causada pelo fungo dimórfico Histoplasma capsulatum, que infecta humanos e uma ampla variedade de mamíferos, incluindo os gatos. No Brasil, somente quatro casos dessa doença foram relatados em gatos domésticos. O presente estudo relata um caso de um gato, vivendo em ambiente domiciliar sem acesso à rua, mas que tinha sido resgatado das ruas havia três anos, que desenvolveu histoplasmose com envolvimento supraorbital. O paciente apresentava espirros, dispneia e aumento dos sons respiratórios e a suspeita clínica foi rinite alérgica. O animal foi tratado com prednisolona e amoxicilina. Com a piora dos sinais clínicos o paciente retornou apresentando um edema na região supraorbital direita, conjuntivite unilateral do olho esquerdo e linfoadenomegalia na região submandibular. A citologia e a cultura revelaram a presença de Histoplasma capsulatum. Inicialmente, itraconazol (10 mg/kg via oral, uma vez ao dia) foi prescrito e devido à piora dos sinais clínicos, anfotericina B (0,5 mg/Kg via subcutânea) foi administrada por duas semanas associada ao triazólico. A terapia com anfotericina B foi descontinuada e o itraconazol mantido por seis meses até a cura clínica e nenhuma recrudescência foi registrada. É importante ressaltar que veterinários devem estar atentos para os sinais clínicos da histoplasmose e que devem considerar a possibilidade de uma infecção subclínica.

Interaction of an opportunistic fungus Purpureocillium lilacinum with human macrophages and dendritic cells.

INTRODUCTION: Purpureocillium lilacinum is emerging as a causal agent of hyalohyphomycosis that is refractory to antifungal drugs; however, the pathogenic mechanisms underlying P. lilacinum infection are not understood. In this study, we investigated the interaction of P. lilacinum conidia with human macrophages and dendritic cells in vitro. METHODS: Spores of a P. lilacinum clinical isolate were obtained by chill-heat shock. Mononuclear cells were isolated from eight healthy individuals. Monocytes were separated by cold aggregation and differentiated into macrophages by incubation for 7 to 10 days at 37°C or into dendritic cells by the addition of the cytokines human granulocyte-macrophage colony stimulating factor and interleukin-4. Conidial suspension was added to the human cells at 1:1, 2:1, and 5:1 (conidia:cells) ratios for 1h, 6h, and 24h, and the infection was evaluated by Giemsa staining and light microscopy. RESULTS: After 1h interaction, P. lilacinum conidia were internalized by human cells and after 6h contact, some conidia became inflated. After 24h interaction, the conidia produced germ tubes and hyphae, leading to the disruption of macrophage and dendritic cell membranes. The infection rate analyzed after 6h incubation of P. lilacinum conidia with cells at 2:1 and 1:1 ratios was 76.5% and 25.5%, respectively, for macrophages and 54.3% and 19.5%, respectively, for cultured dendritic cells. CONCLUSIONS: P. lilacinum conidia are capable of infecting and destroying both macrophages and dendritic cells, clearly demonstrating the ability of this pathogenic fungus to invade human phagocytic cells.

Interaction of an opportunistic fungus Purpureocillium lilacinum with human macrophages and dendritic cells

Introduction Purpureocillium lilacinum is emerging as a causal agent of hyalohyphomycosis that is refractory to antifungal drugs; however, the pathogenic mechanisms underlying P. lilacinum infection are not understood. In this study, we investigated the interaction of P. lilacinum conidia with human macrophages and dendritic cells in vitro. Methods Spores of a P. lilacinum clinical isolate were obtained by chill-heat shock. Mononuclear cells were isolated from eight healthy individuals. Monocytes were separated by cold aggregation and differentiated into macrophages by incubation for 7 to 10 days at 37°C or into dendritic cells by the addition of the cytokines human granulocyte-macrophage colony stimulating factor and interleukin-4. Conidial suspension was added to the human cells at 1:1, 2:1, and 5:1 (conidia:cells) ratios for 1h, 6h, and 24h, and the infection was evaluated by Giemsa staining and light microscopy. Results After 1h interaction, P. lilacinum conidia were internalized by human cells and after 6h contact, some conidia became inflated. After 24h interaction, the conidia produced germ tubes and hyphae, leading to the disruption of macrophage and dendritic cell membranes. The infection rate analyzed after 6h incubation of P. lilacinum conidia with cells at 2:1 and 1:1 ratios was 76.5% and 25.5%, respectively, for macrophages and 54.3% and 19.5%, respectively, for cultured dendritic cells. Conclusions P. lilacinum conidia are capable of infecting and destroying both macrophages and dendritic cells, clearly demonstrating the ability of this pathogenic fungus to invade human phagocytic cells. .

[The importance of the contextual approach in the teaching of biosafety]. / A importância da abordagem contextual no ensino de biossegurança.

Biosafety is a field of knowledge that raises questions geared to genetically modified organisms that are linked to social and job-related employee protection. The educational process involves seeking to create a participative and transforming agent and must therefore transcend the simple concept of teaching. Thus, it is important to contextualize biosafety within a constructive teaching strategy by identification of its core concepts - risk, hazard and accident - which allows each individual to understand how risk is perceived within society and dealt with in academia in order to add multiple skills to tackle the situation. Understanding how the relationship between work and health and its consequences and effects are constructed over the course of time, makes it possible to train more critical and well prepared citizens to participate in decisions of a political and social nature that can influence their future.

A importância da abordagem contextual no ensino de biossegurança / The importance of the contextual approach in the teaching of biosafety

A biossegurança é um campo do conhecimento que insere questões voltadas para organismos geneticamente modificados e relacionadas à proteção social e ocupacional do trabalhador. O seu processo educativo visa à formação de um agente participativo-transformador e, portanto, deve ultrapassar a simples ideia da normalização. Dessa forma, é importante contextualizar a biossegurança dentro de uma estratégia de ensino construtivista com a identificação dos seus conceitos estruturantes - risco, perigo e acidente - que permitam ao indivíduo compreender como o risco é percebido na sociedade e enfocado na academia para, em seguida, agregar múltiplas competências e enfrentá-lo. Conhecer como as relações de trabalho-saúde, suas implicações e impactos vem sendo construídas ao longo do tempo, pode formar um cidadão mais crítico e preparado para participar das decisões de ordem político-social que podem influenciar o seu futuro.

Biosafety is a field of knowledge that raises questions geared to genetically modified organisms that are linked to social and job-related employee protection. The educational process involves seeking to create a participative and transforming agent and must therefore transcend the simple concept of teaching. Thus, it is important to contextualize biosafety within a constructive teaching strategy by identification of its core concepts - risk, hazard and accident - which allows each individual to understand how risk is perceived within society and dealt with in academia in order to add multiple skills to tackle the situation. Understanding how the relationship between work and health and its consequences and effects are constructed over the course of time, makes it possible to train more critical and well prepared citizens to participate in decisions of a political and social nature that can influence their future.

Contaminação fúngica em condimentos de feiras livres e supermercados / Fungal contamination in spices sold in street markets and supermarkets

Quatro tipos de condimentos, Bixa orellana L.(urucum), Cuminum cyminum L. (cominho), Curcumalonga L. (açafrão) e Piper nigrum L. (pimenta-do-reino), provenientes de feiras livres e de supermercados da cidade de Teresina-PI, Brasil, foram avaliados quanto à contaminação por fungos. Das 36 amostras analisadas, 28 (77,77%) apresentaram resultado positivo e 12 espécies fúngicas foram identificadas. Os valores das unidades formadoras de colônias por grama de produto (UFC/g) detectado nas amostras de condimentos comercializados nas feiras livres e industrializados foram maiores do que os do padrão federal de referência, variando de 8,6 × 103 a 1,3 × 105. Não houve diferença significativa entre esses valores. Aspergillus foi o gênero predominante, com seis espécies identificadas, seguido do gênero Penicillium (três espécies). Das amostras de corante, cominho e açafrão, provenientes das feiras livres, nos condimentos foram detectados os maiores valores de UFC/g, respectivamente, para as espécies Exserohilum mcginnisii, Absidia corymbifera e Aspergillus nidulans. Os condimentos analisados neste estudo mostraram a contaminação por fungos potencialmente patogênicos e produtores de micotoxinas, os quais podem constituir um risco para a saúde dos consumidores.

Isolamento de fungos de equipamentos radiográficos odontológicos / Isolation of fungi from dental radiographic equipment

Objetivo: Isolar os fungos dos equipamentos radiográficos dos consultórios odontológicos de Teresina, Piauí, Brasil. Métodos: Foram analisados oito aparelhos de Raios X intrabucal e dois aparelhos de Raios X extrabucal. As coletas foram realizadas nas superfícies dos equipamentos, após procedimentos rotineiros de atendimento a pacientes nos consultórios, por meio de swabs esterilizados, embebidos em salina e friccionados sobre a superfície de cada aparelho. Em seguida, as amostras foram inoculadas em placas de Petri contendo meios Sabourand dextrose agar acrescido de cloranfenicol, incubados à temperatura ambiente e BBLTM CHROMagarTM Candida (BD-Difco, New Jersey, USA), incubados a 37ºC, para o crescimento de fungos filamentosos e leveduras, respectivamente. Resultados: Foram identificadas 17 espécies de fungos e as espécies mais frequentes foram Candida albicans, Aspergillus niger, Cladosporium cladosporioides e Cladosporium oxysporum. Houve predominância, significativa, de fungos na superfície dos componentes do aparelho de Raios X intrabucal quando comparado aos componentes do aparelho de Raios X extrabucal. Conclusão: As espécies fúngicas isoladas das superfícies dos aparelhos radiográficos odontológicos são potencialmente patogênicas e esses equipamentos podem servir de reservatórios ou vetores de fungos representando um risco de aquisição de infecção cruzada para os pacientes, assim como para a equipe odontológica. É necessário que os profissionais da área sejam alertados quanto ao risco inerente ao seu trabalho, treinados e estimulados a aplicarem os princípios de biossegurança para reduzir ou eliminar esses micro-organismos.

Objective: To isolate fungi from radiographic equipment in dentist?s offices in Teresina, Piaui, Brazil. Methods: Eight intraoral and two panoramic X-ray instruments were analyzed. Samples were taken after routine clinical procedures with sterilized swabs followed by inoculation on Petri dishes containing either Sabourand dextrose agar supplemented with chloramphenicol, or BBLTM CHROMagarTM Candida (BD-Difco, New Jersey, USA). The samples were incubated at room temperature, or 37oC, so that filamentous fungi and yeasts, respectively, could develop.Results: Seventeen species of fungi were detected. The most frequent species were Candida albicans, Aspergillus niger, Cladosporium cladosporioides and Cladosporium oxysporum. Additionally, there was significant prevalence of fungi on the surface of intraoral X-ray components as compared to extraoral X-ray components. Conclusion: The species of fungi isolated from the surfaces of dental radiographic equipment in dentist?s offices in Teresina, PI, are potentially pathogenic. These instruments may play a critical role as reservoirs or vectors for fungi. Moreover, they pose a risk of cross-infection to patients and dental staff. Consequently, it is critical to alert professionals in the dentistry area about the inherent risk in their activities and to establish training courses to encourage them to adopt biosafety procedures in order to reduce or eliminate these microorganisms.

Characteristics of Paecilomyces lilacinus infection comparing immunocompetent with immunosuppressed murine model.

The characteristics of Paecilomyces lilacinus infection were evaluated using two murine experimental models: immunocompetent and immunosuppressed. The evaluation criteria for characteristics of infection were clinical signs, weight loss, survival rates, histopathological alterations and the number of viable fungal cells re-isolated from different organs; and those for immunological status were in vitro lymphoproliferative response, cell surface phenotyping and IFN-γ production. Morphological evaluation showed that P. lilacinus isolates presented morphological characteristics consistent with those described in the literature. The immunocompetent mice could be infected by the fungi, but they did not develop the disease, unlike the immunosuppressed mice, which showed clinical signs of mycosis in an environment of suppressed cellular immune response. The hypothesis of latent infection reactivation in mice was not confirmed. The difference observed in the infection rate of the two fungi isolates points to an intrinsic variation between strains of P. lilacinus and led us to hypothesise that even in the presence of immunosuppressed environment, the fungus virulence can play a role in the pathogenesis of hyalohyphomycosis.

Construção do conhecimento em biossegurança: uma revisão da produção acadêmica nacional na área de saúde (1989-2009) / Knowledge building in biosafety: a review of the Brazilian academic production on health themes (1989-2009)

Este artigo refere-se a uma revisão bibliográfica realizada através do portal da Biblioteca Virtual de Saúde com objetivo de identificar os artigos científicos relacionados à biossegurança, publicados em periódicos nacionais, visando a conhecer inicialmente de que forma esse tema vem sendo abordado no Brasil e como está inserido no nexo trabalho e na educação do profissional da área de saúde. Foram identificados inicialmente 126 artigos publicados durante o período de 1989 a outubro de 2009, em periódicos nacionais na área de saúde. Do total de artigos, 46 estavam relacionados diretamente à temática educação, mas apenas oito abordaram a prática educacional em biossegurança. Identificou-se também que a região Sudeste concentrou o maior número de publicação (77 artigos) acerca do tema biossegurança. Apesar da abrangência dos artigos analisados, ainda existe muito que pesquisar sobre o assunto, de forma a ampliar o debate sobre a educação profissional para o setor saúde, onde o trabalhador seja considerado como sujeito da aprendizagem, capaz de ter uma postura crítica-reflexiva do seu ambiente ocupacional.

Genetic characterization of morphologically variant strains of Paracoccidioides brasiliensis.

Molecular characterization of Paracoccidioides brasiliensis variant strains that had been preserved under mineral oil for decades was carried out by random amplified polymorphic DNA analysis (RAPD). On P. brasiliensis variants in the transitional phase and strains with typical morphology, RAPD produced reproducible polymorphic amplification products that differentiated them. A dendrogram based on the generated RAPD patterns placed the 14 P. brasiliensis strains into five groups with similarity coefficients of 72%. A high correlation between the genotypic and phenotypic characteristics of the strains was observed. A 750 bp-RAPD fragment found only in the wild-type phenotype strains was cloned and sequenced. Genetic similarity analysis using BLASTx suggested that this RAPD marker represents a putative domain of a hypothetical flavin-binding monooxygenase (FMO)-like protein of Neurospora crassa.

Genetic characterization of morphologically variant strains of Paracoccidioides brasiliensis

Molecular characterization of Paracoccidioides brasiliensis variant strains that had been preserved under mineral oil for decades was carried out by random amplified polymorphic DNA analysis (RAPD). On P. brasiliensis variants in the transitional phase and strains with typical morphology, RAPD produced reproducible polymorphic amplification products that differentiated them. A dendrogram based on the generated RAPD patterns placed the 14 P. brasiliensis strains into five groups with similarity coefficients of 72 percent. A high correlation between the genotypic and phenotypic characteristics of the strains was observed. A 750 bp-RAPD fragment found only in the wild-type phenotype strains was cloned and sequenced. Genetic similarity analysis using BLASTx suggested that this RAPD marker represents a putative domain of a hypothetical flavin-binding monooxygenase (FMO)-like protein of Neurospora crassa.