Yeasts of the genital region of patients attending the dermatology service at Hospital São Paulo, Brazil / Levaduras de la región genital en pacientes que acuden al servicio de dermatología en el Hospital São Paulo, Brasil

Background. The knowledge of the diversity of yeasts that make up the skin microbiota of human beings is essential for the efficient monitoring of infections to which a person may be predisposed. Aims. This study identified yeasts comprising the genital skin microbiota of patients attending the Dermatology Service at the Hospital São Paulo-UNIFESP, Brazil. Methods. Samples were collected from the genital region of each patient and cultured on Sabouraud dextrose agar. Individual colonies were carefully transferred to tubes daily. Yeasts were identified based on classical methodologies and confirmed using a commercial kit. Results. Eighty-three patients were included in the study. Approximately 80% were women and 20% were men. The average age was 55 years. Hypertension, diabetes, kidney transplant and AIDS were the main underlying diseases reported by the patients. The most prevalent yeasts were Candida parapsilosis (36.1%), Rhodotorula mucilaginosa (9.2%), Rhodotorula glutinis (8.3%), Candida tropicalis (5.5%) and Trichosporon inkin (1.8%). Approximately 78% of the isolates were obtained in pure cultures. Trichosporon inkin was isolated only from women, in contrast to literature describing a high prevalence in males. Conclusions. Our results suggest that Candida albicans is not the main yeast found on genital skin as previously thought, and opportunistic pathogens such as C. parapsilosis, C. tropicalis, Rhodotorula spp. and T. inkin make up the genital skin microbiota, representing a risk for infection in immunocompromised subjects. These results also indicate that women are carriers of T. inkin, the etiological agent of white piedra and trichosporonosis (AU)

Antecedentes. El conocimiento de la diversidad de las levaduras que conforman la microbiota de la piel de los seres humanos es fundamental para un eficaz seguimiento de las infecciones. Objetivos. En este estudio se identificaron las levaduras que componían la microbiota de la piel de los genitales de pacientes que acudieron al Servicio de Dermatología del Hospital São Paulo - Unifesp, Brasil. Métodos. Se recogieron muestras de la región genital de cada paciente y se cultivaron en agar dextrosa Sabouraud. Las colonias individuales se transfirieron cuidadosamente a tubos. La identificación de las levaduras se basó en metodologías clásicas y se confirmó utilizando un kit comercial. Resultados. Ochenta y tres pacientes fueron incluidos en el estudio. Aproximadamente el 80% eran mujeres y el 20% eran hombres. La edad promedio fue de 55 años. La hipertensión, la diabetes, el trasplante renal y el sida fueron las principales enfermedades subyacentes de los pacientes. Las levaduras más frecuentes fueron Candida parapsilosis (36,1%), Rhodotorula mucilaginosa (9,2%), Rhodotorula glutinis (8,3%), Candida tropicalis (5,5%) y Trichosporon inkin (1,8%). Aproximadamente el 78% de los aislamientos crecieron en cultivo puro. Trichosporon inkin fue aislado únicamente en mujeres, en contra de la literatura que describe una alta prevalencia en los hombres. Conclusiones. Nuestros resultados sugieren que Candida albicans no es la principal levadura que se encuentra en la piel de los genitales, como se pensaba, y los agentes patógenos oportunistas C. parapsilosis, C. tropicalis, Rhodotorula spp. y T. inkin conforman la microflora de la piel genital, lo que representa un riesgo inminente de infección para los pacientes inmunocomprometidos. Estos resultados también indican que las mujeres son portadoras del agente etiológico de la piedra blanca y de la tricosporonosis, T. inkin (AU)

Yeasts of the genital region of patients attending the dermatology service at Hospital São Paulo, Brazil.

BACKGROUND: The knowledge of the diversity of yeasts that make up the skin microbiota of human beings is essential for the efficient monitoring of infections to which a person may be predisposed. AIMS: This study identified yeasts comprising the genital skin microbiota of patients attending the Dermatology Service at the Hospital São Paulo-UNIFESP, Brazil. METHODS: Samples were collected from the genital region of each patient and cultured on Sabouraud dextrose agar. Individual colonies were carefully transferred to tubes daily. Yeasts were identified based on classical methodologies and confirmed using a commercial kit. RESULTS: Eighty-three patients were included in the study. Approximately 80% were women and 20% were men. The average age was 55 years. Hypertension, diabetes, kidney transplant and AIDS were the main underlying diseases reported by the patients. The most prevalent yeasts were Candida parapsilosis (36.1%), Rhodotorula mucilaginosa (9.2%), Rhodotorula glutinis (8.3%), Candida tropicalis (5.5%) and Trichosporon inkin (1.8%). Approximately 78% of the isolates were obtained in pure cultures. Trichosporon inkin was isolated only from women, in contrast to literature describing a high prevalence in males. CONCLUSIONS: Our results suggest that Candida albicans is not the main yeast found on genital skin as previously thought, and opportunistic pathogens such as C. parapsilosis, C. tropicalis, Rhodotorula spp. and T. inkin make up the genital skin microbiota, representing a risk for infection in immunocompromised subjects. These results also indicate that women are carriers of T. inkin, the etiological agent of white piedra and trichosporonosis.

Trichosporon inkin: an uncommon agent of scalp white piedra. Report of four cases in Brazilian children.

We report four cases of scalp white piedra (SWP) in Brazilian female children. Morphological and physiological approaches gave inconsistent results for identifying Trichosporon to species level, while the sequencing of the intergenic spacer 1 region of ribosomal DNA accurately identified the agent of SWP as T. inkin. These cases emphasize the occurrence of this species causing this type of infection. The molecular identification of the suspected agent is needed for appropriate epidemiological surveillance of superficial mycoses caused by Trichosporon species.

Onychomycosis and tinea pedis in athletes from the State of Rio Grande Do Sul (Brazil): a cross-sectional study.

Onychomycosis and tinea pedis are common superficial infections caused primarily by dermatophytes. The aim of this investigation was to study the epidemiology, etiological agents, and potential risk factors for infection based on comparison of athletes and non-athletes from a northern region of Rio Grande do Sul (Brazil). Each group consisted of 100 male individuals with ages ranging from 18 to 40 years. After a clinical examination, samples were taken from individuals presenting signs of onychomycosis and/or tinea pedis for direct microscopic examination and culture. Among the athletes, the frequency of onychomycosis and/or tinea pedis was 32%, and for the control group, it was 20%. The athletes presented 16% of onychomycosis, 12% of tinea pedis, and 4% of onychomycosis and tinea pedis together. The distribution in the control group was 10% of onychomycosis, 7% of tinea pedis, and 3% of this association. The pathogens identified were dermatophytes (84.8%) and yeasts (15.2%), and the most commonly identified organism was Trichophyton rubrum, followed by Trichophyton mentagrophytes var. interdigitale. No significant differences were found when the frequency of species distribution in the athletes and non-athlete groups was compared. Risk factors for onychomycosis in athletes included familial cases of fungal infection, contact with domestic animals, and nail trauma, while the risk factors in non-athletes included the habit of not using sandals in public bathrooms and nail trauma. For tinea pedis, the habit of not using sandals in public bathrooms was a predisposing factor in both groups, while hyperhydrosis was a risk factor only in non-athletes. This study concludes that despite the higher number of fungal infections in athletes, there is no significant difference between these groups.

Local phagocytic responses after murine infection with different forms of Fonsecaea pedrosoi and sclerotic bodies originating from an inoculum of conidiogenous cells.

Fonsecaea pedrosoi is an important causative agent of chromoblastomycosis (CBM) especially in humid areas of the world; however, little is known about the infective forms of this agent that cause CBM. The aim of this study was to investigate the murine tissue response to inoculation with different forms of F. pedrosoi and the morphological changes of the fungal cells in vivo. BALB/c mice were inoculated intraperitoneally with hyphae, conidia or conidiogenous cells and conidia (CCC) at a single site. In addition, the abdomen and footpads were infected subcutaneously with CCC. Fungal forms were inoculated at a final concentration of 1 × 10(6) cells. Hyphae and ungerminated conidia inocula could not be transformed into parasitic forms. In tissue, a great number of conidiogenous cells underwent transformation into sclerotic bodies, which were more resistant to phagocytes in vivo than conidia and hyphae. Clinical and mycological cure of animals infected with CCC was observed from the fourth to the sixth week of infection, while conidia and hyphae infections were faster and generally lasted 2 to 3 weeks. A high number of destructed conidia was observed intracellularly in macrophages. The migration of neutrophils to the inflammatory site seems important for microbicidal activity, particularly against hyphae. Our observations suggest that inocula with conidiogenous cells are associated with in vivo transformation into sclerotic bodies and that local immune response involved with host resistance to experimental F. pedrosoi-infection is primarily mediated by neutrophils as observed in histological sections.

Prolonged infection by Fonsecaea pedrosoi after antigenic co-stimulation at different sites in experimental murine chromoblastomycosis.

In the present study, we examined prolonged infection after antigenic co-stimulation by inoculation of the fungus Fonsecaea pedrosoi at two different sites in three mouse strains (BALB/c, Swiss, and C57BL/6). Using this murine model of infection, we showed that antigen induction of infection at more than one site led to a local suppression of active lesions, which increased the time course of experimental chromoblastomycosis (CBM). Footpad infection with a simultaneous infection of the peritoneum or a mucosal site appeared to cause prolonged infection and frequent fungal disseminations. Using knockout (KO) mice, we observed that antigenic co-stimulation caused progressive illness in CD8-KO animals and an effective immune response in the absence of IL-10. In Xid mice, co-stimulation provoked chronic infection (not prolonged), suggesting that B1 B cells play an important role in the control of fungal infection. The tissue response to infection was similar in all co-stimulated mouse groups, as anatomopathologic sections revealed multifocal lesions (granuloma-like). In general, these mice had acute responses at primary antigenic sites with an intense migration of polymorphonuclear leukocytes (PMNs), whereas the distant infection sites (footpad) showed signs of chronic infection. The migration of PMNs to the secondary site (footpad) increased in the later periods of infection, especially after the disappearance of the primary antigenic focus. PMN migration was associated with lesion-dormancy breakage and fungal elimination. Our findings suggest that the host inflammatory/suppression mechanisms induced by antigenic co-stimulation to systemically fight the same pathogen act coordinately through responses that differ at the sites of infection between acute and chronic integrated healing processes that are more prolonged than an acute infection at a single site. However, the long persistence of fungal cells in the host may be linked to microbial adaptation to a parasitic infection as observed in co-stimulated Xid mice.

Antibiosis and dark-pigments secretion by the phytopathogenic and environmental fungal species after interaction in vitro with a Bacillus subtilis isolate

In this work, different reactions in vitro between an environmental bacterial isolate and fungal species were related. The Gram-positive bacteria had terminal and subterminal endospores, presented metabolic characteristics of mesophilic and acidophilic growth, halotolerance, positive to nitrate reduction and enzyme production, as caseinase and catalase. The analysis of partial sequences containing 400 to 700 bases of the 16S ribosomal RNA gene showed identity with the genus Bacillus. However, its identity as B. subtilis was confirmed after analyses of the rpoB, gyrA, and 16S rRNA near-full-length sequences. Strong inhibitory activity of environmental microorganisms, such as Penicillium sp, Aspergillus flavus, A. niger, and phytopathogens, such as Colletotrichum sp, Alternaria alternata, Fusarium solani and F. oxysporum f.sp vasinfectum, was shown on co-cultures with B. subtilis strain, particularly on Sabouraud dextrose agar (SDA) and DNase media. Red and red-ochre color pigments, probably phaeomelanins, were secreted by A. alternata and A. niger respectively after seven days of co-culture.

Na presente investigação, nosso objetivo principal foi relatar diferentes interações in vitro de um isolado bacteriano ambiental com espécies fúngicas. Através da identificação clássica, nós verificamos que o bacilo ambiental apresentava endósporos terminais e subterminais, características metabólicas de mesofilia, acidofilia, halotolerância, redução de nitrato e produção de enzimas, como caseinase e catalase. Análise de seqüências parciais do gene 16S RNAr contendo de 400 a 700 bases revelou identidade com gênero Bacillus. No entanto, a espécie Bacillus subtilis foi confirmada somente depois da análise de seqüências dos genes rpoB, gyrA, and 16S RNAr. Intensa atividade inibitória aos fungos ambientais, como Penicillium sp, Aspergillus flavus, A. niger, e fitopatogênicos, como Colletotrichum sp, Alternaria alternata, Fusarium solani e F. oxysporum f.sp vasinfectum, foi observada em coculturas com a cepa bacteriana (B. subtilis), particularmente em ágar Sabouraud dextrose e ágar DNase. Pigmentos de cor avermelhada e vermelho-amarronzado, provavelmente feomelaninas, foram secretados respectivamente por colônias de A. alternata e A. niger depois de sete dias de co-cultivo.

Onychomycosis in São Paulo, Brazil.

Onychomycosis is a fungal infection of the nails with broad aetiological scope, and it represents 18-40% of all onychopathies and 39% of all superficial mycotic infections. From July 1996 to December 1999, samples of nails were collected from 588 patients with presumptive diagnosis of onychomycosis at the Dermatology and Mycology Divisions EPM\UNIFESP, Brazil, and the diagnosis was confirmed in 247 of these cases. The most common pathogens isolated in this study were yeasts in 52% of positive cultures (Candida albicans 18.3%, Candida parapsilosis 13.8%, other species of Candida 15.4% and other yeasts 4.6%), followed by dermatophytes in 40.6% of positive cultures (the most commonly isolated organisms were Trichophyton rubrum in 33.2%, followed by Trichophyton mentagrophytes in 6.3% and others 1.2%). Non-dermatophyte moulds were isolated in 7.4% of positive cultures (Fusarium spp. 4.5%, Nattrassia mangiferae 2.3% and Aspergillus spp. 0.6%). Distal and lateral subungual onychomycosis (DLSO) was the commonest clinical pattern 44.6% followed by free edge onycholysis (FEO) 38.8% and others. In conclusion, this study demonstrated that T. rubrum is the main agent causing onychomycosis in toenails, and species of genus Candida were the main agents isolated in fingernail onychomycosis in our region.

Correlação entre os resultados laboratoriais e os sinais e sintomas clínicos das pacientes com candidíase vulvovaginal e relevância dos parceiros sexuais na manutenção da infecção em São Paulo, Brasil / Relationship of laboratory results with clinical signs and symptoms of patients with vulvovaginal candidiasis and the significance of the sexual partners for the maintenance of the infection

OBJETIVO: relacionar as leveduras identificadas aos sinais e sintomas clínicos das pacientes com candidíase vulvovaginal e investigar a importância dos parceiros sexuais na reincidência da infecção. MÉTODOS: foi desenvolvido estudo prospectivo de julho de 2001 a julho de 2003 com uma amostra de mulheres residentes na Grande São Paulo. Foram avaliadas 179 pacientes com suspeita clínica de vaginite fúngica, com idade entre 18 e 65 anos. Os critérios para exclusão foram: gravidez, comprometimento imunológico intrínseco e extrínseco, incluindo AIDS, diabetes, imunossupressão, pacientes em terapia com corticosteróides, antibióticos ou hormônios, em pós-menopausa, em uso de dispositivo intra-uterino e duchas vaginais ou espermicidas. Amostras de secreções vaginais ou da glande dos parceiros sexuais de pacientes com vaginite de repetição foram coletadas para microscopia e cultura de fungos. Colônias fúngicas isoladas em CHROMagar Candida foram identificadas por provas clássicas. O teste exato de Fisher foi usado para correlacionar o quadro clínico com as leveduras isoladas das pacientes. RESULTADOS: os sinais e sintomas clínicos mais relevantes na candidíase vulvovaginal foram prurido e corrimento, seguidos por eritema e edema, estatisticamente independente do agente etiológico. Leveduras foram diagnosticadas por microscopia direta em 77 pacientes com vulvovaginites, sendo obtidos 40 cultivos de Candida spp. Candida albicans (70 por cento), C. glabrata (20 por cento), C. tropicalis (7,5 por cento) e C. guilliermondii (2,5 por cento) foram identificadas. As leveduras prevalentes nos parceiros foram C. albicans e C. glabrata. As mesmas espécies foram detectadas nas companheiras e parceiros em 87 por cento dos casos. CONCLUSÕES: as vulvovaginites fúngicas foram mais freqüentes em mulheres entre 18 e 34 anos de idade. Não foi observada correlação entre as espécies de leveduras detectadas e a sintomatologia clínica. Os parceiros sexuais podem ser...

PURPOSE: to relate yeasts identified by laboratory tests to clinical signs and symptoms in patients with vulvovaginal candidiasis, and to investigate the importance of the sexual partners in the recurrence of the infection. METHODS: from July 2001 to July 2003, a sample of 179 patients aged from 18 to 65 years old, with clinical suspicion of fungal vaginitis were analyzed in a prospective study in Great São Paulo. Exclusion criteria included: pregnancy, impaired intrinsic or extrinsic immune response (including Aids), diabetes or immunosuppression; patients undergoing corticosteroid, antibiotic or hormone therapy, in post menopause, with intrauterine device (IUD) or making use of vaginal douches or spemicides. Samples of vaginal and penis secretions from partners of patients with relapse of vaginitis episodes were collected for microscopy and fungal culture. Fungal colonies isolated in CHROMagar Candida were identified by classical methods. Fisher's exact t-test was used to correlate the clinical picture with the yeasts isolated from patients. RESULTS: the most relevant clinical signs and symptoms were pruritus and vaginal discharge followed by erythema and edema, statistically independent from the etiological agent. Direct microscopy revealed yeasts in 77 patients with vulvovaginitis, and 40 Candida spp cultures were obtained. Candida albicans (70 percent), C. glabrata (20 percent), C. tropicalis (7,5 percent) and C. guilliermondii (2,5 percent) were identified. The yeasts prevalent in partners were C. albicans and C. glabrata. The same species were detected in female and male sex partners in 87 percent of the cases. CONCLUSIONS: fungal vulvovaginitis was more frequent in women between 18 and 34 years old. No correlation was observed between the species of yeast detected and clinical symptomatology. Sexual partners are important Candida spp reservoirs and may be related to the maintenance of the vulvovaginal candidiasis.

Microbiologic profile of flexible endoscope disinfection in two Brazilian hospitals

BACKGROUD: Endoscopes are routinely used in hospitals and clinics of the world and they can be potential sources of cross-infection when the decontamination process is unsuitable AIM: The routines of flexible endoscope (bronchoscopes, esophagogastroduodenoscopes and colonoscopes) disinfection procedures used in two Brazilian university hospitals were evaluated during a 3-year period METHODS: Aleatory samples from internal channels of endoscopes were collected after patient examination and after cleaning/disinfection procedures RESULTS: A contamination >3 log10 was achieved in samples recovered from endoscopes after patient examination. These samples yielded gram-negative bacilli (n = 142: 56 percent), gram-positive cocci (n = 43: 17 percent), yeast cells (n = 43: 17 percent), and gram-positive bacilli (n = 26: 10 percent). Approximately, 72 out of 149 samples (48.32 percent) collected after undergoing the cleaning and disinfection procedures disclosed gram-negative bacilli (n = 55: 61 percent), gram-positive cocci (n = 21: 23 percent), gram-positive bacilli (n = 8: 9 percent) and yeast cells (n = 6: 7 percent). Esophagogastroduodenoscopes and colonoscopes were the most frequently contaminated devices. Pseudomonas aeruginosa, Klebsiella pneumoniae, Escherichia coli, Enterobacter spp, Serratia marcescens, Proteus mirabilis, Citrobacter freundii, Staphylococcus aureus, Staphylococcus coagulase negative, Micrococcus luteus, Candida albicans, C. tropicalis, C. glabrata, C. guilliermondii, Bacillus spp and Corynebacterium spp were predominantly identified CONCLUSION: Inappropriate cleaning and low times of disinfection were respectively the major factors associated with the presence of microorganisms in colonoscopes and esophagogastroduodenoscopes. By analyzing the identified germs, hospital disinfection was considered of either intermediate or poor level. After this investigation, both university centers improved their previous protocols for...

RACIONAL: Endoscópios são rotineiramente utilizados em hospitais e clínicas e podem ser fontes potenciais de infecção cruzada quando a descontaminação é inadequada OBJETIVO: As rotinas de descontaminação dos endoscópios flexíveis (broncoscópios, gastrocópios e colonoscópios) realizadas em dois hospitais universitários do Brasil foram avaliadas durante 3 anos MATERIAL E MÉTODOS: Amostras aleatórias foram coletadas dos canais internos dos endoscópios, depois que o aparelho era utilizado nos pacientes e após o processo de desinfecção RESULTADOS: Contaminação superior a 103 foi verificada em amostras coletadas após o exame endoscópico, sendo isolado bacilos gram-negativos (n = 142: 56 por cento), cocos gram-positivos (n = 43: 17 por cento), leveduras (n = 43: 17 por cento) e bacilos gram-positivos (n = 26: 10 por cento). Em 72 das 149 amostras coletadas após procedimentos de limpeza e desinfecção, detectou-se bacilos gram-negativos (n = 55: 61 por cento), cocos gram-positivos (n = 21: 23 por cento), bacilos gram-positivos (n = 8: 9 por cento) e leveduras (n = 6: 7 por cento). Gastroscópios e colonoscópios eram os aparelhos com maior freqüência e taxa de contaminação. Pseudomonas aeruginosa, Klebsiella pneumoniae, Escherichia coli, Enterobacter spp, Serratia marcescens, Proteus mirabilis, Citrobacter freundii, Staphylococcus aureus, Staphylococcus coagulase negative, Micrococcus luteus, Candida albicans, C. tropicalis, C. glabrata, C. guilliermondii, Bacillus spp and Corynebacterium spp foram os mais identificados CONCLUSÃO: A limpeza inapropriada e curto período de tempo de desinfecção eram, respectivamente, os maiores fatores associados com a presença de microrganismos em gastroscópios e colonoscópios. De acordo com os organismos isolados, considera-se que a desinfecção nos hospitais era de nível baixo a intermediário. Após a investigação, os centros de endoscopia adequaram seus protocolos, sanando os problemas verificados nos procedimentos...

Microbiologic profile of flexible endoscope disinfection in two Brazilian hospitals.

BACKGROUND: [corrected] Endoscopes are routinely used in hospitals and clinics of the world and they can be potential sources of cross-infection when the decontamination process is unsuitable AIM: The routines of flexible endoscope (bronchoscopes, esophagogastroduodenoscopes and colonoscopes) disinfection procedures used in two Brazilian university hospitals were evaluated during a 3-year period METHODS: Aleatory samples from internal channels of endoscopes were collected after patient examination and after cleaning/disinfection procedures RESULTS: A contamination >3 log10 was achieved in samples recovered from endoscopes after patient examination. These samples yielded gram-negative bacilli (n = 142: 56%), gram-positive cocci (n = 43: 17%), yeast cells (n = 43: 17%), and gram-positive bacilli (n = 26: 10%). Approximately, 72 out of 149 samples (48.32%) collected after undergoing the cleaning and disinfection procedures disclosed gram-negative bacilli (n = 55: 61%), gram-positive cocci (n = 21: 23%), gram-positive bacilli (n = 8: 9%) and yeast cells (n = 6: 7%). Esophagogastroduodenoscopes and colonoscopes were the most frequently contaminated devices. Pseudomonas aeruginosa, Klebsiella pneumoniae, Escherichia coli, Enterobacter spp, Serratia marcescens, Proteus mirabilis, Citrobacter freundii, Staphylococcus aureus, Staphylococcus coagulase negative, Micrococcus luteus, Candida albicans, C. tropicalis, C. glabrata, C. guilliermondii, Bacillus spp and Corynebacterium spp were predominantly identified CONCLUSION: Inappropriate cleaning and low times of disinfection were respectively the major factors associated with the presence of microorganisms in colonoscopes and esophagogastroduodenoscopes. By analyzing the identified germs, hospital disinfection was considered of either intermediate or poor level. After this investigation, both university centers improved their previous protocols for disinfection and conditions for reprocessing endoscopes.

[Microbiological evaluation of gastroscope decontamination by electrolysed acid water (Clentop WM-1)]. / Análise microbiológica de gastroscópios descontaminados em aparelho Cleantop WM-1 por uso de água eletrolítica ácida.

BACKGROUND: The manual disinfection of endoscopes with glutharaldeyde is widely employed. The great routine in gastroenteroscopy services, low number of equipment and the lack of technical knowledge about the decontamination processes are factors that stimulate the inadequate endoscope disinfection, intensifying the risk of transmission of microorganisms. The electrolysed acid water has been effective in the inactivation and destruction of microorganisms. AIM: The purpose of this investigation was to verify the microbicidal efficiency of electrolyzed acid water (Cleantop WM-1) to decontaminate gastroscopes after their using in patients. MATERIAL AND METHODS: Samples from biopsy channel of flexible endoscopes collected after patient use (n = 20) and after disinfection (n = 20) were cultivated in tryptic soy agar, MacConkey agar and Sabouraud dextrose agar. RESULTS: Seventeen of the 20 samples collected after patients examination yielded gram-negative bacilli, gram-positive coccus and yeast cells in contamination of 3 to 5 log10 ufc/mL. Microbial growth was not verified in samples collected after the decontamination process. Conclusion - In this preliminary study, the mechanical disinfection carried through the Cleantop device with electrolyzed acid water showed satisfactory results for the elimination of microorganisms and time optimization in the reprocessing of gastroscopes.

Análise microbiológica de gastroscópios descontaminados em aparelho Cleantop WM-1 por uso de água eletrolítica ácida / Microbiological evaluation of gastroscope decontamination by electrolysed acid water (Clentop WM-1)

RACIONAL: O método com utilização manual de glutaraldeído é amplamente empregado para desinfecção de endoscópios. A elevada rotina nos serviços de gastroscopia, pequena quantidade de equipamentos e a falta de conhecimento técnico sobre os processos de descontaminação contribuem para desinfecção inadequada dos endoscópios, intensificando o risco de transmissão de microrganismos. A água eletrolítica ácida tem apresentado eficácia na inativação e destruição de microrganismos e vem sendo usada na descontaminação de endoscópios. OBJETIVO: Verificar a eficiência microbicida da água eletrolítica ácida, produzida pelo aparelho Cleantop WM-1, em 20 gastroscópios contaminados após uso em pacientes. MATERIAL E MÉTODOS: Amostras coletadas do canal de biopsia dos endoscópios, após uso em pacientes (n = 20) e depois da desinfecção (n = 20), foram cultivadas em ágar tripticaseína de soja, MacConkey e Sabouraud dextrose. RESULTADOS: Dezessete das 20 amostras coletadas após o uso do aparelho em pacientes revelaram a presença de bacilos gram-negativos, cocos gram-positivos e leveduras em taxas de 103 a 105 ufc/mL. Nenhuma amostra, das 20 coletadas após a descontaminação, apresentou contaminação microbiana. CONCLUSÃO: Nesse estudo preliminar, a desinfecção mecânica realizada pelo aparelho Cleantop com água eletrolítica ácida revelou resultados satisfatórios pela eliminação de microrganismos e otimização no tempo de processamento dos gastroscópios.

Dermatophytosis on the eyelid caused by Microsporum gypseum.

We report a case of tinea on the eyelid in a woman with psoriasis. Due to the history of psoriasis and clinical aspect of the lesion the diagnosis of psoriasis was suggested in the first place; however, laboratory examination revealed Microsporum gypseum. The patient was treated with oral itraconazole resulting in clinical and mycological cure. We emphasize the uncommon location of tinea caused by M. gypseum and the importance of searching for fungal infection on scaly lesions.

Genetic polymorphism of clinical and environmental strains of Pichia anomala.

In this work 20 clinical and 3 environmental yeast isolates were characterized by classical morphological and physiological methods, as well as molecular methods based on PCR of the ITS1-5.8S rDNA-ITS2 region. The characteristic morphology and biochemical profiles observed in these samples correspond to those described for the Pichia genera, more specifically to P. anomala. The profiles of susceptibility to five antifungal drugs were determined by two broth dilution methods. The results obtained by both methods were comparable and showed that clinical isolates presented more resistance to azoles, amphotericin B, and 5-fluorocytosine, than environmental ones did. By amplification and sequencing of internal transcribed spacers (ITS1 and ITS2) and the ribosomal 5.8S DNA, the yeast samples were divided into four groups, where the strains within each group had the same sequence. Of the analyzed yeast isolates, 78% were identified as Pichia anomnala. Using RAPD analysis with seven different Operon primers, polymorphism was observed within the four groups. Our study highlights the growing importance of P. anornala in fungemic episodes in premature neonates. Furthermore, the methodologies used provide a powerful tool to identify and determine differences in similar strains of this yeast.

Genetic polymorphism of clinical and environmental strains of Pichia anomala

In this work 20 clinical and 3 environmental yeast isolates were characterized by classical morphological and physiological methods, as well as molecular methods based on PCR of the ITS1-5.8S rDNA-ITS2 region. The characteristic morphology and biochemical profiles observed in these samples correspond to those described for the Pichia genera, more specifically to P. anomala. The profiles of susceptibility to five antifungal drugs were determined by two broth dilution methods. The results obtained by both methods were comparable and showed that clinical isolates presented more resistance to azoles, amphotericin B, and 5-fluorocytosine, than environmental ones did. By amplification and sequencing of internal transcribed spacers (ITS1 and ITS2) and the ribosomal 5.8S DNA, the yeast samples were divided into four groups, where the strains within each group had the same sequence. Of the analyzed yeast isolates, 78 por percent were identified as Pichia anomala. Using RAPD analysis with seven different Operon primers, polymorphism was observed within the four groups. Our study highlights the growing importance of P. anomala in fungemic episodes in premature neonates. Furthermore, the methodologies used provide a powerful tool to identify and determine differences in similar strains of this yeast.

Fluorescent method for studying the morphogenesis and viability of dermatophyte cells.

The dermatophyte Microsporum canis is commonly isolated from human and animal infection. The morphogenesis of this fungus was studied during its developmental stages through the fluorescent method Fluorescein Diacetate and Ethidium Bromide. To this end, 50 microl dermatophyte suspension were transferred onto cellophane wrapping esterilized discs (2.5 cm of diameter) placed over the surface of Sabouraud dextrose agar on Petri dishes and incubated at 25 degrees C for 30 days. Every 60 minutes during the first 24 hours and every 12 hours for next 29 days, one disc was transferred onto glass slide, covered with equal volumes of freshly prepared fluorescein diacetate (FDA) and ethidium bromide (EB) solution, mounted with a coverslip and incubated in the dark for 30 minutes, at 25 degrees C. Each preparation was then examined on a fluorescent microscope. M. canis presented well defined growth stages: (1) tumescence of cells; (2) germination; (3) development of hyphae; (4) production of conidia and (5) tumescence and formation of arthroconidiae. Using the fluorescent method, non viable cells showed a light bright red coloration and viable cells presented green fluorescence. The principal morphological changes have occurred between the 3rd until the 18th day of culture. The method is very useful to demonstrate the dermatophyte growth stages as well as the perfect differentiation between viable and non viable cells.

Corneal ulcer caused by the new fungal species Sarcopodium oculorum.

We describe a case of keratitis caused by a new species of the hyphomycetous genus Sarcopodium, S. oculorum. The corneal ulcer developed after 5 months of treatment with corticosteroids in a Brazilian boy diagnosed with allergic conjunctivitis. Fungal hyphae and conidia were detected in corneal scrapings, and repeated cultures were positive for this fungus. The infection was resolved with natamycin and ketoconazole. Eleven antifungals were tested against this fungus, and all except flucytosine and fluconazole showed in vitro activity.

Mycetoma caused by Fusarium solani with osteolytic lesions on the hand: case report.

Eumycetoma is a mycotic disease caused by saprophytic soil fungi that are usually inoculated through minor injuries. A case of mycetoma in a Brazilian farmer aged 71 years is reported. This patient presented erythema and edema on the dorsal surface of the left hand with multiple crusted and cicatricial lesions. No macroscopic grains were observed. The histopathological findings showed grains consisted of numerous hyphae which stained well with Gomori-Grocott method. This material obtained by cutaneous biopsy was submitted to culture on Sabouraud's medium and the colonies were identified as Fusarium solani. The radiological studies revealed bone osteolytic lesions and the ultrasound showed pseudocysts and fistulae at the site of this infection. The patient was treated with oral ketoconazole with a good clinical response.

Paracoccidioidomicose em índios brasileiros da tribo Suruí: estudo clínico-laboratorial de 2 casos / Paracoccidioidomycosis in Amerindiam populations of the Brazilian Suruí tribe: a clinical and laboratory study of two cases

Paracoccidioidomicose é considerada a micose sistêmica endêmica mais prevalente na América Latina. Apesar da maior parte da casuística de paracoccidioidomicose ocorrer entre trabalhadores rurais, há poucos casos documentados de ocorrência dessa micose entre índios brasileiros. Säo apresentados 2 casos de paracoccidioidomicose em índios Suruí, família linguística Tupi-Mondé, procedentes de Cacoal, Rondônia. Ambos apresentaram sorologia positiva a imunodifusäo apenas com antígenos da fase miceliana do P. brasiliensis. Os autores apresentam revisäo de literatura sobre a ocorrência dessa micose entre índios brasileiros e discutem a necessidade de futuras investigaçöes buscando caracterizar as diferenças regionais de cepas de P. brasiliensis e seu impacto no diagnóstico sorológico dessa micose