Abstract Sporothrix spp. are the major dimorphic fungus associated with a type of subcutaneous mycosis, sporotrichosis. The limitation of antifungal availability and the past reports of in vitro resistance of Sporothrix spp. clinical isolates makes it important to search for new compounds with antifungal activities. In this study, we therefore evaluate the in vitro activities of complexes coordinated with Co(II) and cobalt chloride hexahydrate against clinical isolates of Sporothrix spp. Broth microdilution test was performed as per M38-A2 from CLSI (2008) in duplicate for 31 clinical isolates of Sporothrix spp. (27 S. brasiliensis e 04 S. schenckii stricto sensu). The antifungal activities of the complexes coordinated with Co(II) and cobalt chloride hexahydrate were detected at a concentration range of 32-128 µg/mL for all isolates. None of the compounds demonstrated any cytotoxicity (to macrophage cells) at the concentration of 200 µg/mL. The activity against Sporothrix spp. recorded in this study instigate the continuity of experimental studies with Co(II) to search for the mechanisms of antifungal action as well as to evaluate its interaction with the commercial antifungal drugs.

Abstract INTRODUCTION: Previous studies that detected Paracoccidioides spp. DNA in soil taken from rural areas have shown this to be a valuable tool for ecological and epidemiological studies. This study reports the detection of Paracoccidioides spp. DNA in soil samples from an urban area of southern Brazil. METHODS: Sixteen soil samples were submitted to nested-PCR and the amplicons of a representative number of positive samples were sequenced. RESULTS: Paracoccidioides spp. DNA was found in 44% of samples. Four DNA amplicons were sequenced, showing 100% homology with P. brasiliensis. CONCLUSIONS: The southern Brazilian urban population is commonly exposed to the Paracoccidioides fungus.

Abstract INTRODUCTION: Several factors can cause false-positive results in the galactomannan (GM) test; however, others remain unknown. Presently, the impact of airborne contamination by Aspergillus conidia during enzyme-linked immunosorbent assay (ELISA) remains uninvestigated. METHODS: We studied 12 A. fumigatus isolates. Fungal conidia were serially diluted and tested for GM detection using the Platelia® Aspergillus enzyme immunoassay (EIA). RESULTS: The conidia concentration required for an EIA-positive result was 4.8 × 103 (median). CONCLUSIONS: This is the first study to evaluate the impact of environmental contamination on the Platelia® Aspergillus EIA assay. Only massive contamination can interfere with GM optical readings, suggesting that environmental contamination does not cause false-positive test results.

Resumo O aumento de casos de esporotricose zoonótica tornou-se um problema de saúde pública em áreas hiperendêmicas para a doença em felinos. O envolvimento dos gatos domésticos nesta transmissão e o contato contínuo destes animais com o solo sugerem que o ambiente possa ser uma importante fonte e/ou reservatório de fungos Sporothrix spp. Assim, este estudo objetivou avaliar a presença de fungos do complexo Sporothrix em amostras de solo provenientes de locais de acesso de felinos infectados no sul do Brasil. Foram coletadas amostras de solo de residências com felinos com esporotricose, de locais próximos a estas residências e também de locais públicos, totalizando 101 amostras. Estas foram processadas pela técnica de plaqueamento direto e incubadas a 25°C por até 15 dias. Dezessete amostras foram paralelamente submetidas a extração de DNA (kitNORGEN BIOTEK CORP®) e técnica de PCR espécie-específico e Nested-PCR. Embora em nenhuma amostra avaliada tenha sido detectada a presença de Sporothrix spp., seja por cultivo ou por biologia molecular, este estudo não descarta o papel do ambiente na tríade do processo infeccioso (solo - animal - humano), sendo necessários outros estudos ampliando área de abrangência, volume, tipo e método de coleta das amostras, bem como técnicas de detecção.

Abstract The increase in zoonotic sporotrichosis cases has become a public health problem in hyperendemic areas for the feline disease. Domestic cats involvement in this transmission and the continuous contact of these animals with soil suggest that the environment may be an essential source and/or reservoir of Sporothrix spp. Thus, the current study aimed to evaluate the presence of Sporothrix spp. in the soil of places where infected cats in Southern Brazil is present. In total, 101 soil samples were collected from the residence of cats with sporotrichosis, from regions around to these residences, and also from public places. These samples were processed using the direct plating technique being incubated at 25 °C for up to 15 days. Additionally, seventeen samples were also submitted to DNA extraction (NORGEN BIOTEK CORP® kit), PCR-specific and Nested-PCR technique. Although the presence of Sporothrix in soil has been undetected, by culture or molecular biology, this study does not reject the role of the environment in the triad of the infectious process (soil-animal-human). Thus, further studies are necessary for expanding the area of coverage, volume, type and method of sampling, and/or detection techniques.

Abstract Vulvovaginal candidiasis (VVC) is an infection of the genital mucosa caused by different species of the genus Candida. Considering the lack of data on this topic in the south of Brazil, this study aimed to assess the prevalence of Candida spp. in the cervical-vaginal mucosa of patients treated at a university hospital in southern Rio Grande do Sul, as well as the etiology and the susceptibility of the isolates against fluconazole, itraconazole, miconazole and nystatin. Samples were collected at the gynecology clinic of the Federal Hospital of the University of Rio Grande, and the isolates were identified using phenotypic and biochemical tests. The susceptibility analysis was performed according to the CLSI M27-A2 protocol. Of the 263 patients included, Candida spp. was isolated in 27%, corresponding to a prevalence of approximately 15% for both VVC and colonization. More than 60% of the isolates were identified as Candida albicans; C. non-albicans was isolated at a rate of 8.6% in symptomatic patients and 14.3% in asymptomatic patients. The prevalence of resistance against fluconazole and itraconazole was 42% and 48%, respectively; the minimal inhibitory concentration of miconazole ranged from 0.031 to 8 µg/mL, and that of nystatin ranged from 2 to >16 µg/mL. The high rate of resistance to triazoles observed in our study suggests the necessity of the association of laboratory exams to clinical diagnosis to minimize the practice of empirical treatments that can contribute to the development of resistance in the isolates.

Resumo A aspergilose caracteriza-se por ser a principal causa de mortalidade de pinguins em cativeiro. A infecção pelo gênero Aspergillus ocorre principalmente por via aérea, porém o fungo pode ter dispersão pela água. Neste sentido, este trabalho teve como objetivo avaliar a água do tanque onde os pinguins-de-Magalhães permanecem para reabilitação no Centro de Recuperação de Animais Marinhos de Rio Grande, RS, Brasil, quanto à presença de fungos filamentosos do gênero Aspergillus. As amostras de água foram coletadas semanalmente durante um período de 10 meses e processadas em um período máximo de seis horas utilizando-se a técnica da membrana filtrante, com incubação a 25 ºC e 37 ºC por até sete dias. Das 40 amostras analisadas, 32 foram positivas para o isolamento do gênero Aspergillus, sendo que dessas 60% pertenciam à espécie A. fumigatus. Algumas variáveis interferiram significativamente no isolamento do gênero Aspergillus e/ou da espécie A. fumigatus, como temperatura de incubação, sazonalidade e densidade populacional. Este trabalho demonstra que Aspergillus spp. está presente na água, podendo essa ser uma potencial fonte de infecção para os pinguins em reabilitação.

Abstract Aspergillosis is the main cause of mortality in captivity penguins. The infection occurs mainly by conidia inhalation of the Aspergillus genus, however, the fungus can also be dispersed by water. Therefore, this study aimed to evaluate water quality of the pool where the rehabilitated penguins remain at Centro de Recuperação de Animais Marinhos in Rio Grande city, Brazil, searching for the presence of the fungus Aspergillus spp. Water samples were collected weekly during a ten-month period and processed within six hours, applying the technique of filtrating membrane, with incubation at 25 ºC and 37 ºC during seven days. Of the forty samples analyzed, thirty-two were positive for the presence of Aspergillus genus, from these 60% correspond to A. fumigatus. Some variables significantly interfered on the isolation of Aspergillus genus and/or Aspergillus fumigatus specie, such as incubation temperature, seasonality and population density. This study showed Aspergillus spp. is present in the water, being one of the possible sources of infections for penguins in rehabilitation.

Paracoccidioides brasiliensis is the etiological agent of the major systemic mycosis in Brazil, called paracoccidioidomycosis. Although the Rio Grande do Sul is considered an endemic area of the disease, there are few studies on the ecology of P. brasiliensis in the state. Therefore, this study aimed to evaluate the infection of P. brasiliensis in horses from the mesoregion of Southwest Riograndense, using these animals as sentinels. Serological techniques, such as double immunodiffusion in agar gel (AGID) and indirect ELISA, were performed to detect the anti-gp43 P. brasiliensis antibody in horses from five different farms in the region of Bagé, RS, Brazil. Serology was performed in 200 Pure Blood English horses up to two years of age that were born and raised exclusively at the farms. Of these horses, 12% had anti-gp43 antibodies according to the ELISA results, with rates ranging from 0 to 30% according to the farm of origin (p < 0.001). Based on the immunodiffusion results, all equine serum samples were negative. These results indicate the presence of the fungus P. brasiliensis in the middle region of the southwestern state of Rio Grande do Sul, Brazil.

RESUMO A criptococose é uma micose primariamente pulmonar, sendo adquirida por meio de inalação de propágulos fúngicos infectantes encontrados no ambiente. Um dos principais agentes etiológicos da criptococose é a espécie Cryptococcus neoformans , que apresenta distribuição mundial. O objetivo desse trabalho foi pesquisar a ocorrência de Cryptococcus spp. em excretas de Columba livia (pombos domésticos) encontradas na área externa do prédio do Hospital Universitário Dr. Miguel Riet Correa Junior, centro de referência para o tratamento da Síndrome da Imunodeficiência Adquirida na região sul do estado do Rio Grande do Sul, Brasil. Durante um período de dez meses, foi coletado um total de 40 amostras de excretas secas da área acadêmica e da área hospitalar do Hospital Universitário. As excretas foram adicionadas de solução salina com cloranfenicol, homogeneizados em vórtex, semeados em ágar Níger e incubados a 25ºC com observações diárias até sete dias. A identificação do micro-organismo foi realizada por provas fenotípicas e bioquímicas. Das 40 amostras processadas e analisadas, 13 (32,5%) foram positivas para o isolamento de Cryptococcus spp., variando de 20.000 a 3.000.000 UFC/g de fezes. O isolamento desse fungo em um ambiente hospitalar é relevante em saúde pública, pois evidencia a exposição dos indivíduos que frequentam esse local a propágulos infectantes.

ABSTRACT Cryptococcosis is primarily a pulmonary mycosis, being acquired by inhalation of infective fungal propagules found in the environment. One of the main etiological agents of cryptococcosis is the yeast Cryptococcus neoformans , which has worldwide distribution and whose naturalhabitat is the feces of birds. The aim of this study was to investigate the occurrence of Cryptococcus spp. in excreta ofColumba livia (domestic pigeon) found in the outer area of the building of Hospital Universitário Dr. Miguel Riet Correa Junior, a reference center for the treatment of Acquired Immunodeficiency Syndrome in southern state of Rio Grande do Sul, Brazil. During a period of ten months, a total of 40 samples of dried excreta was collected from the site's academic area and hospital. To the excreta, were added saline solution with chloramphenicol, homogenized in a vortex, cultivated in Niger seed agar and incubated at 25°C with daily observations up to seven days. The identification of the microorganism was performed by phenotypical and biochemical tests. From the 40 samples processed and analyzed, 13 (32.5%) were positive for the isolation ofCryptococcus spp., with the quantity of yeasts ranging from 20.000 to 3.000.000 CFU/g of feces. Isolation of the opportunistic fungusCryptococcus spp. at a university hospital is relevant in public health for demonstrating the exposure of individuals who frequent this site to infectant propagules.

BACKGROUND: Onychomycosis or nail fungal infection is the most common nail disease. Despite the wide range of studies on this condition, it remains difficult to establish the correct diagnosis and effective treatment. OBJECTIVES: To evaluate the efficacy of classical laboratory methods for the diagnosis of onychomycosis, and the in vitro susceptibility of the its main etiological agent to antifungals used in routine. METHODS: Nail samples of 100 patients with clinically suspected feet onychomycosis were collected to confirm the diagnosis by direct mycological examination and fungal culture. In vitro antifungal susceptibility testing was performed against strains of the main dermatophyte isolated by microdilution, according to the standardized protocol (M38-A2 - CLSI) RESULTS: Clinical diagnosis of onychomycosis was confirmed by laboratory analysis in 59% of patients. Of these, 54.2% were positive only in direct mycological examination, 44.1% in direct mycological examination and culture, and one case (1.7%) was positive only in culture, resulting in weak agreement between these tests (Kappa = 0.385; p <0.001) High minimum inhibitory concentration values of fluconazole and itraconazole were observed in 66.7% and 25.0% of isolates of T. rubrum tested. Additionally, high MIC values of terbinafine and ciclopirox was detected in only one isolate, and this was one of the strains in which in vitro activity of itraconazole and fluconazole has not been proven. CONCLUSIONS: Poor agreement was observed between direct mycological examination and culture for the diagnosis of onychomycosis, with direct mycological examination being significantly more sensitive. Except for fluconazole, the other three antifungals tested showed good in vitro activity against clinical isolates of T. rubrum.

O estudo objetivou investigar a liberação in vitro de galactomanana (GM) em distintas espécies patogênicas de fungos do gênero Aspergillus. Grande variabilidade foi detectada tanto intra quanto inter espécies, sendo as cepas da espécie A. terreus relacionadas aos maiores índices de GM detectados. O significado in vivo destes achados permanece em aberto, porém merece investigação.

Here we investigate the extent to which different Aspergillus species release galactomannan (GM) in vitro. Marked variability was observed in GM reactivity between and within Aspergillus species, with A. terreus strains showing the highest GM indexes. The in vivo significance of these findings remains to be determined.

Sporotrichosis occurs after fungal implantation of Sporothrix spp. in the skin, and is the main subcutaneous mycosis in Latin America. Here we describe three atypical cases of the disease. The first case report an extra-cutaneous occurrence of the disease with joint infection; the second one describes a patient with bilateral lymphocutaneous form of sporotrichosis; and the third shows a zoonotic cutaneous case with the development of an erythema nodosum as a hypersensitivity reaction. These cases show the disease importance on the region and the necessity of fungal culture to the diagnosis confirmation.

The oral microbiota of humans and animals is made up of a wide variety of yeasts and bacteria, but microbiota of dogs is not totally described. Although such identification is an important step to establish the etiopathogenesis and adequate therapy for the periodontal disease The aim of this study was to evaluate and correlate oral alterations with the presence of yeasts in oral cavity of female dogs. After clinical evaluation samples from healthy and from dogs with oral diseases were obtained from three different oral sites by swabs, curettes, millimeter periodontal probes and HA membrane tip in cellulose ester. Yeast identification was performed through macroscopic and microscopic colony features and biochemical tests. Dental calculus was the most prevalent occurrence in the oral cavity of 59 females. However, the isolation of yeasts was significantly higher (p < 0.05) in animals suffering from halitosis. Eleven yeast species were identified, namely: Malassezia pachydermatis, Rhodotorula spp., Candida albicans, C. catenulata, C. famata, C. guilliermondii, C. parapsilosis, C. intermedia, Trichosporon asahii, T. mucoides and Cryptococcus albidus. It could be concluded that the yeasts are part of the microbiota from the different sites of the oral cavity of the female canines studied without causing any significant alterations except halitosis.

Os fungos filamentosos são oportunistas de fagócitos, motivo pelo qual aspergilose é incomum em pacientes com Aids. A apresentação clínica depende do estado imune, tamanho do inóculo fúngico e doença de base. São relatados neste trabalho seis casos de bola fúngica em pacientes com Aids. Neste grupo, todos tiveram tuberculose como doença de base e hemoptise foi o principal sintoma. O diagnóstico da bola fúngica foi através da apresentação clínica, achados radiológicos combinados com imunodifusão radial dupla, exame micológico direto e cultivo do material do trato respiratório, sendo A. fumigatus o agente isolado

Aspergillus is a phagocyte opportunistic fungus that causes aspergillosis, an unusual disease in patients with AIDS. Six cases of fungal ball in patients with AIDS are reported here. In this group, all patients had hemoptysis and tuberculosis as the underlying lung disease. The diagnosis of pulmonary fungus ball was based on the clinical and radiographic feature, combined with serological and mycological evidence of Aspergillus fumigatus.

Piperacillin-tazobactam is a broad spectrum antimicrobial agent that can cause false-positive results in the commercial Platelia Aspergillus EIA test. So far, no study has been performed in Latin America to evaluate the clinical implication of this finding. Here we studied the potential for galactomannan detection in piperacillin-tazobactam batches commercialized in the Brazilian market. Five batches from distinct laboratories were tested in duplicate in the Platelia Aspergillus EIA according to the manufacturer's instructions. Only one drug showed crossreaction at a cut-off of 0.5. Human serum was spiked with this particular drug aiming to mimic achievable piperacillin-tazobactam concentrations in the serum. Results were all negative for galactomannan detection, even at high drug concentrations. Results from this pilot study suggest that piperacillin-tazobactam might not be a clinically significant cause of false-positive results in the Platelia Aspergillus EIA test in Brazil.