ABSTRACT A 49-year-old patient with changes in the nails of the hallux for 10 years was diagnosed with onychomycosis. The identity of the causative agent was confirmed as Cladosporium halotolerans from the Cladosporium sphaerospermum species complex using molecular techniques. MALDI-TOF identified the agent as C. sphaerospermum complex species. Overall, species such as onychomycosis agents should attract special attention to avoid mistakes in the identification process while considering a probable contaminant as responsible for the disease. These species deserve attention since there are rare descriptions of them as causes of onychomycosis. It is important to recognize them as causes of disease and not just as a probable contaminant.

The aim of this study was to identify Candida species isolated from women diagnosed with recurrent vulvovaginal candidiasis (RVVC) and their partners; and to evaluate the fluconazole (FLZ) susceptibility of the isolates. In a period of six years, among 172 patients diagnosed with vulvovaginal candidiasis, 13 women that presented RVVC and their partners were selected for this investigation. The isolates were obtained using Chromagar Candida medium, the species identification was performed by phenotypic and molecular methods and FLZ susceptibility was evaluated by E-test. Among 26 strains we identified 14 Candida albicans, six Candida duobushaemulonii, four Candida glabrata, and two Candida tropicalis. Agreement of the isolated species occurred in 100% of the couples. FLZ low susceptibility was observed for all isolates of C. duobushaemulonii (minimal inhibitory concentration values from 8-> 64 µg/mL), two C. glabrata isolates were FLZ-resistant and all C. albicans and C. tropicalis isolates were FLZ-susceptible. This report emphasises the importance of accurate identification of the fungal agents by a reliable molecular technique in RVVC episodes besides the lower antifungal susceptibility profile of this rare pathogen C. duobushaemulonii to FLZ.

OBJETIVO: Identificar as espécies predominantes e o papel dos parceiros sexuais na manutenção das vulvovaginites recorrentes por Candida spp. MÉTODOS: Entre agosto de 2007 e março de 2012, foi efetuado estudo prospectivo de 830 pacientes com idades variáveis entre 18 e 65 anos e vaginites fúngicas. Foram excluídos pacientes com diabetes mellitus, síndrome da imunodeficiência adquirida (AIDS), em uso de corticoterapia, antibioticoterapia, hormonoterapia, imunossupressão, duchas vaginais, dispositivos intrauterinos (DIUs) ou espermicidas. A identificação das espécies de Candida foi feita por métodos fenotípicos e genotípicos. O teste do χ2 foi usado para correlacionar a Candida spp. nos parceiros masculinos e a recorrência nas vaginites. RESULTADOS: O agente fúngico foi isolado em um total de 40 mulheres, sendo 24 com vaginites recorrentes, e em 15 dos seus parceiros sexuais, dos quais 10 eram assintomáticos, e 5, sintomáticos. Houve concordância das espécies encontradas no casal em 100% das recorrências. C. albicans (62,4 e 60%), C. glabrata (29,1 e 33,3%) e C. guilliermondii foram as espécies identificadas. Candida tropicalis (4,1%) foi isolada de apenas uma paciente. Nas 16 mulheres restantes que tinham vaginites não complicadas, C. albicans foi isolada em todas. C. glabrata foi isolada em apenas dois de seus parceiros assintomáticos. CONCLUSÃO: Houve predominância de C. albicans; parceiros sintomáticos ou assintomáticos podem ter papel importante como reservatório e fonte de transmissão de leveduras, principalmente nos quadros de vulvovaginites recorrentes.

PURPOSE: To identify the predominant species and the role of sexual partners in the maintenance of recurrent vulvovaginitis by Candida spp. METHODS: A prospective study of 830 patients aged 18 to 65 years with yeast vaginitis was performed between August 2007 and March 2012. Patients with diabetes mellitus, AIDS or taking corticosteroids, antibiotics or hormone therapy and immunosuppressed patients, patients using vaginal douches, spermicides or intrauterine devices were excluded from the study. Candida species were identified by phenotypic and genotypic methods. The chi-square test was used to correlate the presence of Candida spp. in male partners with the recurrence of vaginitis. RESULTS: The fungal agent was isolated from a total of 40 women, 24 with recurrent vaginitis and from 15 of their sexual partners, 10 of whom were asymptomatic while 5 were symptomatic. There was agreement of the species found in the couple in 100% of recurrences. C. albicans (62.4 and 60%), C. glabrata (29.1 and 33.3%) and C. guilliermondii species were identified. Candida tropicalis (4.1%) was isolated from only one patient. Candida albicans was isolated from the remaining 16 women who had uncomplicated vaginitis. C. glabrata was isolated from only two of the asymptomatic partners. CONCLUSION: There was a predominance of C. albicans and symptomatic or asymptomatic partners can play an important role as a reservoir and source of transmission of yeast, especially in cases of recurrent vulvovaginitis.

OBJETIVO: Avaliar a eficácia do uso tópico de iodo-povidona 0,5% em ceratite experimental por Fusarium solani em coelhos. MÉTODOS: Ceratite fúngica por Fusarium solani foi induzida no olho direito de 24 coelhos da raça New Zealand. Os coelhos foram divididos aleatoriamente em 3 diferentes grupos de tratamento: Grupo I (iodo-povidona) - tratados com iodo-povidona 0,5%; Grupo II (natamicina) - tratados com natamicina 5%; Grupo III (controle) - tratados com solução salina. Os coelhos dos 3 grupos foram tratados por 3 dias e sacrificados em seguida. As córneas foram removidas cirurgicamente, maceradas e incubadas em meio BHI. Semeou-se culturas em placas de ágar Sabouraud, diariamente, durante 7 dias, e contou-se o número de unidades formadoras de colônias (UFC). Os coelhos foram avaliados clinicamente durante o período de tratamento. RESULTADOS: Os grupos iodo-povidona e natamicina demonstraram melhor eficácia do que o grupo controle considerando-se o número de coelhos nos quais não houve crescimento de colônias. Entretanto, não houve diferença estatística significante entre os 3 grupos quando se analizou o número de UFC (p>0,05). CONCLUSÃO: Este estudo demonstrou considerações metodológicas importantes na utilização de modelos animais para o teste de agentes antifúngicos. Usando a metodologia de contar UFC e com este tamanho amostral, administração tópica de iodo-povidona 0,5% não demonstrou benefício do tratamento de ceratite fúngica experimental causada por Fusarium solani quando comparado com a administração tópica de natamicina 5%.

PURPOSE: To evaluate the efficacy of topical administration of 0.5% povidone-iodine in experimental Fusarium solani keratitis in rabbits. METHODS: Fungal keratitis caused by Fusarium solani was induced in the right eye of 24 New Zealand rabbits. The rabbits were randomly divided into 3 different treatment groups: Group I (povidone-iodine) - treated with topical 0.5% povidone-iodine; Group II (natamycin) - treated with topical 5% natamycin; and Group III (control) - treated with topical saline solution. In all groups the rabbits were treated for three days and then sacrificed. The corneas were excised, macerated and immersed in 10 mL BHI. Culture samples were plated daily on Sabouraud's agar for 7 days, and the number of colony-forming units (CFU) was counted. The rabbits were clinically evaluated during the treatment period. RESULTS: The povidone-iodine and natamycin groups demonstrated better efficacy than the control group based on the number of rabbits with no colonies growing. However, there were no statistically significant differences between the three groups when the number of CFU was analyzed (p>0.05). CONCLUSIONS: Our study demonstrates important methodological considerations in the use of in vivo animal models for the testing of antifungal agents. Using this sample size and methodology of counting CFU, topical 0.5% povidone-iodine demonstrated no benefit in the treatment of experimental Fusarium solani when compared with topical 5% natamycin.

OBJETIVOS: Determinar a micobiota de conjuntiva sadia em indivíduos diabéticos, segundo tipo de diabetes, idade, sexo, tempo de doença, tipo de tratamento e estádio da retinopatia. Estabelecer a micobiota anemófila nas salas de colheita. MÉTODOS: Estudo transversal de 803 diabéticos residentes na zona urbana de São Paulo - SP/Brasil. Foi usado para primo-isolamento o meio de cultivo ágar Sabouraud dextrose com cloranfenicol e para identificação dos fungos filamentosos a chave de De Hoog. RESULTADOS: Dos diabéticos avaliados, 6,6% (53/803) apresentavam diabetes tipo 1 e 93,4% (750/803) tipo 2. Os cultivos positivos para fungos em conjuntiva de diabéticos foi 4,2% (34/803), sendo 1,9% (1/53) nos diabéticos tipo 1 e 4,4% (33/740) nos diabetes tipo 2 (p=0,720). Não foi verificada associação estatisticamente significante quanto à presença ou não de isolamentos de fungos em relação idade (p=0,575), sexo (p=0,517), tempo de doença (p=0,633), tipo de tratamento (p=0,422) e estádio de retinopatia diabética (p=0,655) desses indivíduos. Todos os fungos identificados foram filamentosos: Aspergillus spp. representou 59,5% (25/42) dos isolamentos sendo 47,6% (20/42) Aspergillus niger. Ocorreu crescimento de fungos anemófilos do ar ambiente da sala, observando-se coincidências entre as espécies isoladas no ar e na conjuntiva. CCONCLUSÕES: Foi identificada presença de micobiota em conjuntiva sadia de diabéticos, não havendo associação entre a maior positividade de isolamentos fúngicos e o tipo de diabetes, idade, sexo, tempo de doença, tipo de tratamento e estádio da retinopatia diabética. Nas salas de colheita foi identificada micobiota anemófila.

PURPOSE: To determine the mycobiota of the healthy conjunctiva in diabetic individuals, according to diabetes type, age, sex, disease time, type of treatment, and stage of diabetic retinopathy of the individuals. To identify the anemophilus mycobiota in the sampling rooms. METHODS: A cross-sectional study was carried out on 803 diabetics who reside in the urban area of São Paulo-SP/Brazil. Sabouraud's dextrose agar culture with chloramphenicol was used for primoisolation, and the key of De Hoog was used to identify filamentous fungi. RESULTS: Of the evaluated diabetics, 6.6% (53/803) presented type 1 diabetes and 93.4% (750/803) type 2. The positive cultures for fungi in the conjunctiva of diabetics was 4.2% (34/803), with 1.9% (1/53) in type 1 diabetics and 4.4% (33/740) in type 2 diabetics (p=0.720). With respect to the presence or not of isolated fungi, there was no statistically significant association regarding age (p=0.575), sex (p=0.517), disease time (p=0.633), type of treatment (p=0.422), and diabetic retinopathy stage (p=0.655) of the tested individuals. The identified fungi were all filamentous: Aspergillus spp. represented 59.5% (25/42) of isolations and 47.6% (20/42) of isolated species were Aspergillus niger. Growth of anemophilus fungi occurred in the air of the room and coincidences were observed between the isolated species from the air and those from the conjunctiva. CONCLUSIONS: Presence of mycobiota in healthy conjunctivas of diabetics was identified, with no significant association between the greater number of positive fungi isolations and the type of diabetes, age, sex, disease type, type of treatment, and stage of diabetic retinopathy. In the collection rooms, anemophilus mycobiota was identified.

Objetivo: Apresentar a freqüência e o tipo de fungos identi-ficados de infecções corneanas. Métodos: Levantamento retrospectivo dos casos de ceratites micóticas, no Laboratório de Microbiologia Ocular do Departamento de Oftalmologia da Universidade Federal de São Paulo (UNIFESP) no período entre 1995 a 1998. Descrição dos isolamentos de fungos, análise dos fatores desencadeantes e relação com o número de ceratites infecciosas no mesmo período. Resultados/Conclusão: Ceratites micóticas foram diagnos-ticadas em 61 (5,48%) dos 1113 pacientes que apresentaram úlcera de córnea de etiologia infecciosa, com variação de 3,46-9,25%, ao ano. Fungos filamentosos foram identificados em 47 casos (77,04%) e leveduras em 14 (22,95%). Fusarium foi o gênero mais freqüente (50,82%), seguido de Candida (22,95%) e Aspergillus (8,19%). Foram também isolados fungos raros como agentes etiológicos de ceratites como: Phaeosiaria sp; Phoma sp; Fonsecaea pedrosoi e Exserohilum rostratum.

Purpose: To present the frequency and type of identified fungi from infectious keratitis. Methods: Retrospective survey of the cases of mycotic keratitis in the period from 1995 to 1998, at the Laboratory of Ocular Microbiology of the Department of Ophthalmology of the Federal University of São Paulo. Description of the fungal isolations, analysis of the causative factors and relation to the number of infectious keratitis in the same period. Results/Conclusion: Mycotic keratitis was diagnosed in 61 (5.48%) of the 1,113 patients who presented ulcer of the cornea of infectious etiology, ranging from 3.4 to 9.25%, per year. Filamentous fungi were identified in 47 cases (77.04%) and yeasts in 14 (22.95%). Fusarium was the most frequent genus (50.82%), followed by Candida (22.95%) and Aspergillus (8.19%). Phaeosiaria sp, Phoma sp, Fonsecaea pedrosoi, Exserohilum rostratum, that are rare etiological fungal agents of keratitis, were also isolated.

RESUMO Foi estudada a variação sazonal da microbiota fúngica da conjuntiva de 194 pacientes HIV soro-positivos, na cidade de São Paulo. Os cultivos apresentaram positividade de 40,0 e 52,0%, respectivamente, no verão e outono e de 20% no inverno e 20,8% na primavera. Aspergillus spp foi o fungo mais isolado no verão, inverno e primavera, enquanto que no outono foi o Penicillium spp.

SUMMARY The seasonal variation of fungal microbiota of the conjunctiva of 194 assymptomatic and symptomatic HIV positive patients, in the city of São Paulo, was studied. In summer, the cultures were positive in 40,0%, in fall 52,0%, in winter 20,0% and in spring 20,8%. Aspergillus spp, was the most frequent fungus found in summer, winter and spring while Penicillium spp was the most frequent one in fall.

RESUMO Foi realizado estudo experimental com 21 coelhos para avaliar a eficácia da sulfadiazina de prata a 1 % no tratamento da ceratite por Fusarium solani. Os animais foram divididos em três grupos tratados com sulfadiazina de prata a 1 % (grupo S); com veículo (grupo V) e sem tratamento (grupo C). Todos os animais apresentaram cura clínica, com tempo médio igual à 17 dias no grupo C; 17 dias no grupo V e 20 dias no grupo S. A secreção ocular esteve presente por um período médio de 8 dias no grupo C; 7 dias no grupo V e 5 dias no grupo S. A conjuntivite esteve presente em todos os animais desaparecendo em 13 dias no grupo C; 14 no "V"e 12 no "S". Observou-se melhora da doença epitelial em 5 dias, 4 e 3 dias, respectivamente, nos grupos C, V e S. A formação de abscesso ocorreu em todos os animais, tendo desaparecido mais precocemente no grupo S. A vascularização corneana foi maior nos animais tratados com sulfadiazina de prata. Não houve diferença estatisticamente significante entre os achados. O estudo histopatológico mostrou processo inflamatório crônico, vascularização e fibrose localizada. Conclui-se que a sulfadiazina de prata a 1 %, usada topicamente mostrou-se ineficaz no tratamento das ceratites por F. solani em coelhos.

SUMMARY An experimental sutdy was done with 21 rabbits to evaluate the silver sulphadiazine 1% efficacy in the treatment of Fusarium solani keratitis. They were divided in three groups treated with silver sulfadiazine 1% (group S); vehicle (group V) and without treatment (group C). All animais had clinic cure diagnoses in an average of 17 days in group C; 17 days in group V and 20 days in group S. Ocular secretion was present during 8 days in group C; 7 days in group V and 5 days in group S. Conjunctivitis had disappeared in 13 days in group C; 14 days in Vand 12 days in group S. The epithelial keratitis was better in 5 days; 4 days and 3 days, respectively, in groups C, V and S. The abscess was present in ali animais and disappeared early in group Ss. The cornealvascularization was more intensive in those animaIs treated with silver sulphadiazine. There was no significant statistic difJerence in the results. Histopathologic studies indicated chronic inflammatory process and vascularization. Topical silver sulphadiazine 1% wwas in efficient in the treatment of experimental F. solani keratitis.