Bacillus subtilis induces morphological changes in Fonsecaea pedrosoi in vitro resulting in more resistant fungal forms in vivo

Interactions among microorganisms may be the cause of morphological modifications, particularly in fungal cells. The aim of this work was to examine the changes that occur in cells of the fungus Fonsecaea pedrosoi after in vitro co-culturing with Bacillus subtilis and to explore the results of this interaction in vivo in an experimental murine infection. B. subtilis strain was inoculated into a 15-day pure culture of F. pedrosoi. In vitro, after 48 hours of co-culturing, the fungal cells were roundish. The secretion of fungal dark pigments and production of terminal chlamydoconidia were observed in hyphae after one week. In the in vivo study, two animal groups of 30 BALB/c mice each were employed. One group was inoculated intraperitoneally with hyphal fragments from the co-culture of bacteria and fungi; the other group was infected only with F. pedrosoi hyphae. After seven days of infection, both animal groups developed neutrophilic abscesses. Phagocytosis of bacilli by macrophages occurred at three days. At later periods, generally after 25 days, only roundish cells similar to sclerotic bodies remained in the tissues while hyphae were eliminated by 15 to 20 days. These fungal forms originated mainly from terminal chlamydoconidia. The co-culturing between bacteria and fungi may constitute a mechanism to rapidly obtain resistant fungal forms for host defenses, especially for chromoblastomycosis (CBM) experimental infections.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)FAPEMATFederal University of Mato Grosso Department of Basic Health SciencesFederal University of São Paulo Department of Microbiology, Immunology and ParasitologyUNIFESP, Department of Microbiology, Immunology and ParasitologyFAPEMAT: 002.138/2007SciEL

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.This work received the financial support of CNPq and FAPEMAT project number Nº. 002.138/2007

Ocorrência de microsporum gypseum no tegumento de bovino São (São Jose do rio preto, São Paulo, Brasil)

Se describe por primera vez, en Brasil, la presencia de Microsporum gypseum en piel de bovino sano. Se llama la atención sobre la importancia de esos animales como reservório de M. gypseum

Paracoccidioidomycosis in Amerindian populations of the Brazilian Suruí tribe: a clinical and laboratory study of two cases

Paracoccidioidomycosis has been considered the most frequent endemic systemic mycosis in Latin America. Although most cases of paracoccidioidomycosis involve rural workers, this systemic fungal disease has been scarcely reported among Amerindian populations from Brazil. We report two cases of paracoccidioidomycosis in Tupi-Mondé Amerindians from Cacoal, state of Rondônia, Brazil. Both cases exhibited positive serological results by a specific immunodiffusion test only when the assay was performed with antigens obtained from the mycelial form of P. brasiliensis. The authors present a literature review of paracoccidioidomycosis in Brazilian Amerindians and discuss the need for further investigations about the impact of the antigenic diversity of P. brasiliensis from different geographic areas on the serological diagnosis of PCM.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. São apresentados 2 casos de paracoccidioidomicose em índios Suruí, família linguística Tupi-Mondé, procedentes de Cacoal, Rondônia. Ambos apresentaram sorologia positiva à imunodifusão apenas com antígenos da fase miceliana do P. brasiliensis. Os autores apresentam revisão de literatura sobre a ocorrência dessa micose entre índios brasileiros e discutem a necessidade de futuras investigações buscando caracterizar as diferenças regionais de cepas de P. brasiliensis e seu impacto no diagnóstico sorológico dessa micose.Universidade Federal de São Paulo (UNIFESP)UNIFESPSciEL

Efeito tópico de iodo-povidona 0,5% comparado com natamicina 5% em ceratites fúngicas causadas por Fusarium solani em um modelo animal de coelhos: estudo piloto

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.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%.Universidade Federal de São Paulo (UNIFESP)University of CaliforniaUNIFESPSciEL