RESUMO Avaliou-se a susceptibilidade de Sporothrix albicans(n = 1), S. brasiliensis(n = 1), S. globosa(n = 1), S. mexicana(n = 1) e S. schenckii(n = 36) frente à terbinafina (TRB) e a TRB em combinação com itraconazol (ITZ), cetoconazol (KTZ) e voriconazol (VRZ) pelo método de microdiluição ( checkerboard); o perfil enzimático destas espécies foi também avaliado, com base no kit API-ZYM. A maioria das interações foram aditivas (27,5%, 32,5% e 5%) ou indiferentes (70%, 50% e 52,5%) para TRB+KTZ, TRB+ITZ e TRB+VRZ, respectivamente. Antagonismo foi observado em 42,5% dos isolados para a combinação TRB+VRZ. Com base nos perfis enzimáticos, as cepas de Sporothrix schenckii evidenciaram 14 biotipos distintos. A atividade da leucina arilamidase (LA) só foi observada em S. albicanse S. mexicana. As espécies S. globosae S. mexicanaforam as únicas que não evidenciaram atividade da enzima β-glucosidase (GS). Estes resultados poderão contribuir para um melhor entendimento da virulência e resistência entre as espécies do gênero Sporothrixem futuros estudos.

SUMMARY Sporothrix schenckiiwas reclassified as a complex encompassing six cryptic species, which calls for the reassessment of clinical and epidemiological data of these new species. We evaluated the susceptibility of Sporothrix albicans (n = 1) , S. brasiliensis (n = 6) , S. globosa (n = 1), S. mexicana(n = 1) and S. schenckii(n = 36) to terbinafine (TRB) alone and in combination with itraconazole (ITZ), ketoconazole (KTZ), and voriconazole (VRZ) by a checkerboard microdilution method and determined the enzymatic profile of these species with the API-ZYM kit. Most interactions were additive (27.5%, 32.5% and 5%) or indifferent (70%, 50% and 52.5%) for TRB+KTZ, TRB+ITZ and TRB+VRZ, respectively. Antagonisms were observed in 42.5% of isolates for the TRB+VRZ combination. Based on enzymatic profiling, the Sporothrix schenckii strains were categorized into 14 biotypes. Leucine arylamidase (LA) activity was observed only for S. albicans and S. mexicana. The species S. globosaand S. mexicanawere the only species without β-glucosidase (GS) activity. Our results may contribute to a better understanding of virulence and resistance among species of the genus Sporothrixin further studies.

In vitro interaction between tacrolimus (FK506) and four azoles (fluconazole, ketoconazole, itraconazole and voriconazole) against thirty clinical isolates of both fluconazole susceptible and -resistant Candida glabrata were evaluated by the checkerboard microdilution method. Synergistic, indifferent or antagonism interactions were found for combinations of the antifungal agents and FK506. A larger synergistic effect was observed for the combinations of FK506 with itraconazole and voriconazole (43%), followed by that of the combination with ketoconazole (37%), against fluconazole-susceptible isolates. For fluconazole-resistant C. glabrata, a higher synergistic effect was obtained from FK506 combined with ketoconazole (77%), itraconazole (73%), voriconazole (63%) and fluconazole (60%). The synergisms that we observed in vitro, notably against fluconazole-resistant C. glabrata isolates, are promising and warrant further analysis of their applications in experimental in vivo studies.

Malassezia pachydermatisis associated with dermatomycoses and otomycosis in dogs and cats. This study compared the susceptibility of M. pachydermatis isolates from sick (G1) and healthy (G2) animals to azole and polyene antifungals using the M27-A3 protocol. Isolates from G1 animals were less sensitive to amphotericin B, nystatin, fluconazole, clotrimazole and miconazole.

C. dubliniensis é uma levedura oportunista que, embora já tenha sido isolada de vários sítios anatômicos é, com maior frequência, encontrada na boca de pacientes infectados pelo HIV. Embora tenham sido realizados numerosos estudos sobre a epidemiologia e filogenia, seus fatores de virulência como atividade exoenzimática e atividade hemolítica, são, ainda, pouco conhecidos. Neste estudo comparou-se a atividade in vitro de proteinase, hialuronidase, condroitin sulfatase e atividade hemolítica de 18 cultivos de C. dubliniensis com 30 cultivos de C. albicans, todos isolados de pacientes com SIDA. Foi evidenciada maior atividade de proteinase em C. albicans em relação a C. dubliniensis (p < 0,05). Todos os isolados de C. dubliniensis evidenciaram atividade de hialuronidase e condroitin-sulfatase de forma similar ao observado com C. albicans (p > 0,05). Constatou-se que a atividade hemolítica foi influenciada pelo CaCl2; em sua ausência não foram observadas diferenças na atividade hemolítica das duas espécies; todavia, ao se agregar 2,5% de CaCl2, a atividade hemolítica de C. dubliniensis foi reduzida enquanto a de C. albicans, estimulada (p < 0,05).

Candida dubliniensis is an opportunistic yeast that has been recovered from several body sites in many populations; it is most often recovered from the oral cavities of human immunodeficiency virus-infected patients. Although extensive studies on epidemiology and phylogeny of C. dubliniensis have been performed, little is known about virulence factors such as exoenzymatic and hemolytic activities. In this study we compared proteinase, hyaluronidase, chondroitin sulphatase and hemolytic activities in 18 C. dubliniensis and 30 C. albicans strains isolated from AIDS patients. C. albicans isolates produced higher amounts of proteinase than C. dubliniensis (p < 0.05). All the tested C. dubliniensis strains expressed hyaluronidase and chondroitin sulphatase activities, but none of them were significantly different from those observed with C. albicans (p > 0.05). Hemolytic activity was affected by CaCl2; when this component was absent, we did not notice any significant difference between C. albicans and C. dubliniensis hemolytic activities. On the contrary, when we added 2.5 g% CaCl2, the hemolytic activity was reduced on C. dubliniensis and stimulated on C. albicans tested strains (p < 0.05).

O presente estudo teve como objetivo comparar o ágar suco de tomate, um tradicional meio utilizado para observação de ascósporos em leveduras, com o ágar semente de niger, ágar caseína e ágar semente de girassol, na diferenciação fenotípica entre C. albicans e C. dubliniensis. Após 48 h de incubação a 30 ºC, os 26 isolados de C. dublinienis (100%) evidenciaram a formação de clamidoconídios igualmente em todos os meios comparados. Entretanto, quando semeados com C. albicans, a formação de clamidoconídios foi raramente observada, resultando nos seguintes percentuais de ausência destas estruturas: ágar suco de tomate (92,47%), ágar niger (96,7%), ágar caseína (91,39%), ágar semente de girassol (96,7%). Estes resultados permitem-nos sugerir a utilização do ágar suco de tomate como mais um meio que, já no primo-isolamento, é capaz de, presuntivamente, diferenciar C. albicans de C. dubliniensis.

The purpose of the present study is to compare the tomato juice agar, a well known medium employed to observe ascospore formation, with niger seed agar, casein agar and sunflower seed agar, applied to a differentiation between C. dubliniensis and C. albicans. After 48 hours of incubation at 30 ºC all 26 (100%) C. dubliniensis isolates tested produced chlamydospores on tomato juice agar as well as in the other three media evaluated. However, when we inoculated all media with C. albicans, the absence of chlamydospores became resulting in the following percents: tomato juice agar (92.47%), niger seed agar (96.7%), casein agar (91.39%), and sunflower seed agar (96.7%). These results indicate that tomato juice agar is another medium which can also be used in the first phenotypic differentiation between C. dubliniensis and C. albicans.

Avaliou-se a capacidade do ágar suco de tomate (ágar V8) em diferenciar Candida dubliniensis de Candida albicans com base na produção de clamidoconídios. Noventa e três isolados de Candida albicans e vinte e seis de Candida dubliniensis foram incluídos; 100% de Candida dubliniensis formaram clamidoconídios e 92,5% de Candida albicans não evidenciaram estas estruturas. Estes resultados permitem sugerir este meio como recurso alternativo na identificação presuntiva de Candida dubliniensis.

We evaluated the capacity of tomato juice agar (V8 agar) to differentiate Candida dubliniensis from Candida albicans based on chlamydospore production. Candida albicans (n= 93) and Candida dubliniensis (n= 26) were studied; 100% of Candida dubliniensis showed chlamydospores and in 92.5% of Candida albicans isolates these elements were absent. These results suggest this medium as an alternative tool for presumptive differentiation between these species.