Candida duobushaemulonii: an emerging rare pathogenic yeast isolated from recurrent vulvovaginal candidiasis in Brazil.

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.

A peculiar case of onychomycosis caused by Cladosporium halotolerans.

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.

A peculiar case of onychomycosis caused by Cladosporium halotolerans

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.

[Study of mycobiota in the healthy conjunctiva of diabetics who reside in the urban area of the city of São Paulo, Brazil]. / Estudo da micobiota em conjuntiva sadia de diabéticos, residentes na área urbana da cidade de São Paulo-Brasil.

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.

[The role of the symptomatic and asymptomatic sexual partners in the recurrent vulvovaginitis]. / O papel dos parceiros sexuais sintomáticos e assintomáticos nas 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.

Effects of temperature and incubation time on the in vitro expression of proteases, phospholipases, lipases and DNases by different species of Trichosporon.

Fungi produce a broad spectrum of enzymes capable of degrading different substrates in nature. When the substrate is the tissue of a vertebrate host, these enzymes acts as a fungal virulence factor that increases the pathogenicity of the fungus. Trichosporon yeasts are emerging pathogens that infect immunocompromised patients. Little is known about the virulence characteristics of these fungi. The aim of this research was to characterize the behavior of protease, phospholipase, lipase and DNase production in different species of Trichosporon, with a focus on the influence of incubation temperature on the expression of these enzymes. Classical methodologies were used in all of the experiments, and the results were statistically analyzed. The proportions of the samples that produced each type of enzyme were as follows: lipases (95.5%), phospholipases (56.8%), proteases (50,0%) and DNases (38.6%). The incubation temperature was an important factor in the expression of enzymatic activity, and it influenced the incubation period of each species. Although these data concerning the enzymatic activity expressed by isolates of Trichosporon are valuable, further research is warranted to completely characterize this new pathogen, as well as in vivo studies to determine the roles of these enzymes in the pathogenesis of trichosporonosis.

O papel dos parceiros sexuais sintomáticos e assintomáticos nas vulvovaginites recorrentes / The role of the symptomatic and asymptomatic sexual partners in the recurrent vulvovaginitis

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. .

Effect of topical 0.5% povidone-iodine compared to 5% natamycin in fungal keratitis caused by Fusarium solani in a rabbit model: a pilot study.

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.

Limitations of DNA sequencing for diagnosis of a mixed infection by two fungi, Phaeoacremonium venezuelense and a Plectophomella sp., in a transplant recipient.

We describe a rare case of a subcutaneous infection by both Phaeoacremonium venezuelense and Plectophomella sp. in a Brazilian male. Sequencing of a beta-tubulin gene fragment allowed us to confirm the identification of the former. However, a similar procedure of sequencing rRNA gene fragments was not useful for the identification of the latter fungus.

Effect of topical 0.5 percent povidone-iodine compared to 5 percent natamycin in fungal keratitis caused by Fusarium solani in a rabbit model: a pilot study / Efeito tópico de iodo-povidona 0, 5 por cento comparado com natamicina 5 por cento 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 percent 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 percent povidone-iodine; Group II (natamycin) - treated with topical 5 percent 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 percent povidone-iodine demonstrated no benefit in the treatment of experimental Fusarium solani when compared with topical 5 percent natamycin.

OBJETIVO: Avaliar a eficácia do uso tópico de iodo-povidona 0,5 por cento 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 por cento; Grupo II (natamicina) - tratados com natamicina 5 por cento; 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 por cento não demonstrou benefício do tratamento de ceratite fúngica experimental causada por Fusarium solani quando comparado com a administração tópica de natamicina 5 por cento.

Estudo da micobiota em conjuntiva sadia de diabéticos, residentes na área urbana da cidade de São Paulo - Brasil / Study of mycobiota in the healthy conjunctiva of diabetics who reside in the urban area of the city of São Paulo, Brazil

OBJETIVOS: Determinar a micobiota de conjuntiva sadia em indivíduos diabéticos, segundo tipo de diabetes, idade, sexo, tempo de doenca, 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 identificacão dos fungos filamentosos a chave de De Hoog. RESULTADOS: Dos diabéticos avaliados, 6,6 por cento (53/803) apresentavam diabetes tipo 1 e 93,4 por cento (750/803) tipo 2. Os cultivos positivos para fungos em conjuntiva de diabéticos foi 4,2 por cento (34/803), sendo 1,9 por cento (1/53) nos diabéticos tipo 1 e 4,4 por cento (33/740) nos diabetes tipo 2 (p=0,720). Não foi verificada associacão estatisticamente significante quanto à presenca ou não de isolamentos de fungos em relacão idade (p=0,575), sexo (p=0,517), tempo de doenca (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 por cento (25/42) dos isolamentos sendo 47,6 por cento (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 presenca de micobiota em conjuntiva sadia de diabéticos, não havendo associacão entre a maior positividade de isolamentos fúngicos e o tipo de diabetes, idade, sexo, tempo de doenca, tipo de tratamento e estádio da retinopatia diabética. Nas salas de colheita foi identificada micobiota anemófila.

Análise laboratorial de ceratites fungicas em Serviço Universitário / Laboratorial analyses of fungal keratitis in a University Service

Apresentar a frequencia e o tipo de fungos identificados de infecçöes corneanas. Métodos: Levantamento retrospectivo dos casos de ceratites micóticas, no Laboratório Micribiologia Ocular do Departamento de Oftalmologia da Universidade Federal de Säo Paulo (UNIFESP) no período entre 1995 a 1998. Descriçäo dos isolamentos de fungos, análises dos fatores desencadeantes e relaçäo com o número de ceratites infecciosas no mesmo período. Resultados/Conclusäo: Ceratites micóticas foram diagnosticados em 61 (5,48 por cento) dos 113 pacientes que apresentaram úlcera de córnea de etiologia infecciosa, com variaçäo de 3,46 - 9,25 por cento, ao ano. Fungos filamentosos foram identificados em 47 casos (77,04 por cento) e leveduras em 14 (22,95 por cento). Fusarium foi o gênero mais frequente (50,82 por cento), seguido de Candida (22,95 por cento) e Aspergillus (8,19 por cento). Foram também isolados fungos raros com agentes etiológicos de ceratite como: Phaeosiaria sp; Phoma sp; Fonsecaea pedrosoi e Exserohilum rostratum.

Ocorrência de Microsporum gypseum no tegumento de bovino säo (Säo José do Rio Preto, Säo Paulo, Brasil) / Occurrence of Microsporum gypseum in the tegument of a healthy cattle (Säo José do Rio Preto, Säo 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

Candida albicans isoladas de pacientes com SIDA: sensibilidade "in vitro" aos agentes antifúngicos / Candida albicans from AIDS patients: susceptibility \"in vitro\" to antifungal agents

Resumo: Por meio da determinaçäo da CIM(Concentraçäo Inibitória Mínima) e da CFM(Concentraçäo Fungicoda Mínima), os autores verificaram a sensibilidade de candida albicans isoladas de pacientes com SIDA, frente a Anfotericina B, Nistatina, Cetoconazol e miconazol.Os autores näo verificaram a ocorrência de amostras resistente e discutem o fenômeno da resistência de C.albicans aos antifúngicos poliênicos e imidazólicos.

Estudo experimental da eficácia da sulfadiazina de prata a 1 por cento no tratamento da ceratite por Fusarium solani / Experimental study of silver sulphadiazine 1 per cents in the treatment of Fusarium solani

Foi realizado estudo experimental com 21 coelhos para avaliar a eficácia da sulfadiazina de prata a 1 por cento no tratamento da ceratite por Fusarium solani. Os animais foram divididos em 3 grupos tratados com sulfadiazina de prata a 1 por cento (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 secreçäo 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 formaçäo de abscesso ocorreu em todos os animais, tendo desaparecido mais precocemente no grupo S. A vascularizaçäo corneana foi maior nos animais tratados com sulfadiazina de prata. Näo houve diferença estatisticamente significante entre os achados. O estudo histopatológico mostrou processo inflamatório crônico, vascularizaçäo e fibrose localizada. Conclui-se que a sulfadiazina de prata a 1 por cento, usada topicamente mostrou-se ineficaz no tratamento das ceratites por F. solani em coelhos

Variaçäo sazonal da microbiota fúngica da conjuntiva de pacientes HIV soro-positivos / Seasonal variation of the fungal microbiota in the conjunctiva of HIV soropositive patients

Foi estudada a variaçäo sazonal da microbiota fúngica da conjuntiva de 194 pacientes HIV positivos, na cidade de Säo Paulo. Os cultivos apresentaram positividade de 40,0 e 52,0 por cento, respectivamente, no veräo e outono e de 20 por cento no inverno e 20,8 por cento na primavera. Aspergillus spp foi o fungo mais isolado no veräo, inverno e primavera, enquanto que no outono foi o Penicillium spp