Molecular detection of Paracoccidioides in soil from an urban area of southern Brazil

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.

Can Aspergillus fumigatus conidia cause false-positive results in the galactomannan enzyme immunoassay test?

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.

Epidemiologia de micoses, pitiose e micotoxicoses em equinos no sudeste do Rio Grande do Sul / Epidemiology of mycoses, pitiosis and micotoxicosis in horses in southeastern Rio Grande do Sul, Brazil

Descrevem-se os aspectos epidemiológicos das doenças causadas por fungos e oomicetos na população de equinos na região sudeste do Rio Grande do Sul, estabelecendo as taxas epidemiológicas, suas causas e sua importância sanitária na região. Foi realizada a epidemiologia descritiva por meio do cálculo da incidência das doenças encontradas ao longo dos anos e verificada a existência de associação entre a ocorrência dessas enfermidades e o sexo, a raça e a estação do ano. Entre os anos de 1978 e 2014 a pitiose teve prevalência de 49,71% (86/173), as micotoxicoses 30,05% (52/173), sendo 45 casos de leucoencefalomalácia e sete de ergotismo. As micoses tiveram prevalência de 19,65% (34/173), sendo as dermatofitoses as mais prevalentes com 58,82% (20/34) dos casos. As espécies de dermatófitos mais frequentemente isoladas foram Trichophyton mentagrophytes 60% (12/20), Trichophyton equinum 25% (5/20) e Microsporum gypseum, Microsporum canis e Trichophyton verrucosum ambos responsáveis por 5% (1/20) das infecções. Rinosporidiose foi diagnosticada em 35,29% (12/34) dos casos. Micoses uterinas causadas por Candida albicans e Cryptococcus laurentii foram observadas em 5,88% (2/34) dos casos. Alergia por Cladosporium sp. teve um registro. De acordo com as incidências calculadas entre 1990 e 2014 a pitiose teve incidência mediana (IM) 2,98 e distância interquartil (DI) =3,82, as fêmeas tiveram chance 4,18 vezes maiores de desenvolver a doença, a enfermidade ocorre independente das estações climáticas. A leucoencefalomalácia teve IM=0,0; DI 1,00 e equinos machos tiveram 3,4 vezes mais chance de desenvolver a doença que fêmeas, no inverno a possibilidade de ocorrência dessa enfermidade foi seis vezes maior. O ergotismo teve IM = 0,00; DI = 0,000, rinosporidiose IM=0,00; DI=0,088 e dermatofitose IM=0,00; DI=0,935. A pitiose foi mais prevalente entre as doenças encontradas, podendo ser considerada endêmica na região. Considera-se que a magnitude das doenças possa ser ainda maior dentro do rebanho equino, uma vez que as doenças descritas não são de notificação obrigatória e algumas são bem conhecidas por veterinários e proprietários, que muitas vezes não fazem a confirmação laboratorial do diagnóstico.(AU)

The epidemiological aspects of diseases caused by fungi and oomycetes in horses in southeastern Rio Grande do Sul, Brazil, is described. The epidemiological rates, their causes, and health importance in the region were established. A descriptive epidemiology study was carried out in relation of potential risk factors. The impact on these diseases in the region was measured. From 1978 to 2014, pythiosis had a prevalence of 49.71% (86/173), and mycotoxicoses of 30.05% (52/173), with 45 cases of leukoencephalomalacia and 7 of ergotism. The prevalence of fungal infections was 19.65% (34/173) of cases. Dermatophytosis was the most prevalent fungal infection with 58.82% (20/34) of cases. The most isolated dermatophyte species were Trichophyton mentagrophytes 60% (12/20), Trichophyton equinum 25% (5/20) and Microsporum gypseum, Microsporum canis, and Trichophyton verrucosum, both responsible for 5% (1/20) of infections. Rhinosporidose was diagnosed in 35.29% (12/34) of cases. Uterine mycosis caused by Candida albicans and Cryptococcus laurentii was observed at 5.88% (2/34) of cases. Cladosporium sp. allergy was noted in one record. According to the incidence from 1990 to 2014, pythiosis had median incidence (MI) of 2.98 and interquartile range (DI) of 3.82. Mares were 4.18 times likely to develop the disease then males. The disease occurs in the region in every season. Leukoencephalomalacia had MI of 0.0; DI 1.00 and male horses were 3.4 times more likely than mares to develop the disease. Leukoencephalomalacia was 6 times more likely to occur during winter. Ergotism had MI of 0.00; DI of 0.000, rhinosporidiosis MI of 0.00, DI of 0.088 and ringworm MI of 0.00, and DI of 0.935. In the study pythiosis had the highest prevalence among the diseases observed, and may be considered endemic in the region. The magnitude of the diseases observed may be even greater within the equine herd, since these diseases are not of obligatory notification and some are well known by veterinarians and owners, who often do not obtain a laboratory confirmation of the diagnosis.(AU)

Prevalence of Candida spp. in cervical-vaginal samples and the in vitro susceptibility of isolates

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.

Cytotoxicity and anti-Sporothrix brasiliensis activity of the Origanum majorana Linn. oil

Abstract The study aimed to evaluate the anti-Sporothrix sp. activity of the essential oil of Origanum majorana Linn. (marjoram), its chemical analysis, and its cytotoxic activity. A total of 18 fungal isolates of Sporothrix brasiliensis (n: 17) from humans, dogs and cats, and a standard strain of Sporothrix schenckii (n: 1) were tested using the broth microdilution technique (Clinical and Laboratory Standard Institute - CLSI M27-A3) and the results were expressed in minimal inhibitory concentration (MIC) and minimal fungicidal concentration (MFC). The MIC50 and MIC90 of itraconazole against S. brasiliensis were 2 µg/mL and 8 µg/mL, respectively, and the MFC50 and MFC90 were 2 µg/mL and >16 µg/mL, respectively, with three S. brasiliensis isolates resistant to antifungal. S. schenckii was sensitive at MIC of 1 µg/mL and MFC of 8 µg/mL. For the oil of O. majorana L., all isolates were susceptible to MIC of ≤2.25-9 mg/mL and MFC of ≤2.25-18 mg/mL. The MIC50 and MIC90 were ≤2.25 mg/mL and 4.5 mg/mL, respectively, and the MFC50/90 values were twice more than the MIC. Twenty-two compounds were identified by gas chromatography with a flame ionization detector (CG-FID) and 1,8-cineole and 4-terpineol were the majority. Through the colorimetric (MTT) assay, the toxicity was observed in 70-80% of VERO cells between 0.078 and 5 mg/mL. For the first time, the study demonstrated the satisfactory in vitro anti-Sporothrix sp. activity of marjoram oil and further studies are needed to ensure its safe and effective use.

Anti- Sporothrix spp. activity of medicinal plants

ABSTRACT Cases of sporotrichosis in humans and animals without satisfactory clinical response have increased, a warning sign of strains resistant to conventional antifungal agents. The urgent search for alternative therapies was an incentive for research on medicinal plants with anti-Sporothrix spp. properties. A bibliographic survey was performed based on scientific papers about in vitro and in vivo antifungal activity of essential oils and extracts of plants in differents solvents against the fungal of the Sporothrix schenckii complex. The study methodology consisted of a literature review in Google Scholar, Science Direct, Pubmed, Bireme and Springer link with papers from 1986 to 2015. We found 141 species of plants that were investigated, of which 100 species were concentrated in 39 botanical families that had confirmed anti-Sporothrix activity. Combretaceae, Asteraceae and Lamiaceae represented the botanical families with the greatest number of plants species with antifungal potential, using different methodologies. However, there are few studies with medicinal plants in experimental infection in animals that prove their activity in the treatment of sporotrichosis. It reinforces the need for further research related to standardization of in vitro methodologies and in vivo studies related to safety and to toxicity potential of these plants with anti-Sporothrix spp. activity.

RESUMO Casos de esporotricose em humanos e animais sem resposta clínica satisfatória têm aumentado, sinal de alarme para o surgimento de cepas resistentes aos antifúngicos convencionais. A urgente busca por alternativas terapêuticas tem incentivado as pesquisas em plantas medicinais com atividade anti-Sporothrix spp. Um levantamento bibliográfico foi realizado com base em artigos científicos sobre a atividade antifúngica in vitro e in vivo de óleos essenciais e extratos de plantas preparados em diferentes solventes contra o complexo Sporothrix schenckii. A metodologia do estudo consistiu em uma revisão bibliográfica em Google Scholar, Science Direct, Pubmed, Bireme e Springer link com artigos desde 1986 até 2015. Foram encontradas 141 espécies de plantas já investigadas, das quais 100 espécies concentradas em 39 famílias botânicas apresentaram atividade anti-Sporothrix spp. confirmada. Combretaceae, Asteraceae e Lamiaceae representaram as famílias botânicas com maior número de espécies vegetais com potencial antifúngico, empregando diferentes metodologias. Entretanto, há poucos estudos com plantas medicinais em infecção experimental animal comprovando sua atividade no tratamento da esporotricose. Reforça-se a necessidade de mais pesquisas relacionadas à padronização de metodologias in vitro e a estudos in vivo relacionados à segurança e potencial tóxico dessas plantas com atividade anti-Sporothrix spp.

Seroepidemiology of Paracoccidioides brasiliensis infection in horses from Rio Grande do Sul, Brazil

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.

Serological monitoring of antibodies for an early diagnosis ofaspergillosis in captive penguins / Monitoramento sorológico com detecção de anticorpos para diagnóstico precoce da aspergilose em pinguins em cativeiro

This study aimed to evaluate the efficacy of detection of anti-Aspergillus fumigatus antibodies in captive penguins by double radial agar gel immunodiffusion (AGID) for the aspergillosis diagnosis. We included 134 Magellanic penguins (Spheniscus magellanicus) in rehabilitation at the Center for Recovery of Marine Animals (CRAM / FURG). All of them were monitored by AGID weekly until its final destination (death or release), totalizing 660 serum samples studied. All animals were clinically accompanied and post-mortem examinations was performed in penguins that died during the studied period. A total of 28% (37/134) of the penguins died, 89.2% (33/37) due to aspergillosis, 11% (4/37) by other causes and 97 were released. From the 33 animals with proven aspergillosis, 21 presented anti- A. fumigatus antibodies by AGID, being the average interval between death and positive AGID 16.4 days. Twelve animals with negative serology died of aspergillosis. The sensitivity and specificity rates were 63.6% and 95% respectively, and the positive and negative predictive values were 80.7% and 88.9% respectively. These data demonstrate that the serological monitoring for detection of antibodies by AGID can be an important tool for the diagnosis of aspergillosis in penguins.

Este estudo teve como objetivo avaliar a eficácia da detecção de anticorpos anti- Aspergillus fumigatus em pinguins em cativeiro por imunodifusão radial dupla em gel de ágar (IDGA) para diagnóstico da aspergilose. Foram incluídos 134 pingüins de Magalhães (Spheniscus magellanicus) em reabilitação no Centro de Recuperação de Animais Marinhos (CRAM/FURG), que foram monitoradas por IDGA, semanalmente, até o seu destino final (morte ou de liberação), totalizando 660 amostras de soro estudadas. Todos os animais foram acompanhados clinicamente e exames post mortem foram realizados em pingüins que vieram a óbito durante o período de estudo. Um total de 28% (37/134) dos pinguins foram a óbito, 89,2% (33/37) de aspergilose, 11% (4/37) de outras causas, e 97 foram liberados. A partir dos 33 animais com aspergilose comprovada, 21 apresentaram anticorpos anti- A. fumigatus por IDGA, sendo o intervalo médio entre a morte e IDGA positivas 16,4 dias. Doze animais com sorologia negativa vieram a óbito por aspergilose. As taxas de sensibilidade e especificidade foram de 63,6% e 95%, respectivamente, e os valores preditivos positivos e negativos foram de 80,7% e 88,9 %, respectivamente. Estes dados demonstram que o monitoramento sorológico para detecção de anticorpos por IDGA pode ser uma ferramenta importante no diagnóstico de aspergilose em pinguins.

Susceptibility of dermatophytic fungi to commonly used disinfectants / Suscetibilidade de fungos dermatófitos a desinfetantes comumente utilizados

This study aimed evaluate the antidermatophytic activity of three commercial disinfectants commonly used for environmental control of microorganisms in veterinary medicine. Sodium hypochlorite at 40 µL/mL, chloro-phenol derived at 30 µL/mL and chlorhexidine digluconate at 66.7 µL/mL were tested against 14 strains of dermatophytes, identified as Microsporum canis (n: 3) and Microsporum gypseum (n: 11). The tests was performed in accordance with guidelines of the Clinical and Laboratory Standards Institute (CLSI), documents M38-A2 and M51-A, adapted to disinfectants. In the microdilution broth test, chlorhexidine digluconate had MIC values (Minimum Inhibitory Concentration) of 4.16 µL/mL and MCF (Minimum Fungicidal Concentration) from 4.16 to 8.33 µL/mL, while chloro-phenol derived obtained MIC and MCF of 1.87 µL/mL, and both disinfectants had fungicidal activity at concentrations below the recommended. Sodium hypochlorite obtained MIC from 10 to 80 µL/mL and MFC of 40 to 80 µL/mL, requiring at most isolates twice the recommended concentration to achieve same activity. In the disc diffusion test, the mean inhibition zones for chlorhexidine digluconate was 10.53 mm, for chloro-phenol of 9.9 mm and for sodium hypochlorite was 6.2 mm. Chlorhexidine digluconate and chloro-phenol presented a significant reduction in the growth of dermatophytes, while sodium hypochlorite in concentration recommended showed a low antifungal activity against tested isolates.

O objetivo do estudo foi avaliar a atividade antidermatofítica de três desinfetantes comerciais frequentemente utilizados no controle ambiental de micro-organismos em medicina veterinária. Hipoclorito de sódio a 40 µL/mL, derivado de clorofenol a 30 µL/mL e digluconato de clorexidine a 66.7 µL/mL foram testados contra 14 cepas de dermatófitos, identificados como Microsporum canis (n: 3) e Microsporum gypseum (n: 11). Foram utilizadas as diretrizes do Clinical and Laboratory Standard Institute (CLSI), documentos M38-A2 e M51-A, com adaptações para desinfetantes. Na microdiluição em caldo, digluconato de clorexidine apresentou valores de CIM (Concentração Inibitória Mínima) de 4.16 µL/mL e CFM (Concentração Fungicida Mínima) entre 4.16 a 8.33 µL/mL; derivado de clorofenol obteve CIM e CFM de 1.87 µL/mL, demonstrando que ambos os desinfetantes apresentaram atividade fungicida em concentrações inferiores às recomendadas. O hipoclorito de sódio demonstrou CIM entre 10 a 80 µL/mL e CFM de 40 a 80 µL/mL, requerendo duas vezes a concentração recomendada pelo fabricante para obter atividade fungicida frente a maioria dos isolados fúngicos testados. No teste de disco-difusão, a média das zonas de inibição do digluconato de clorexidine foi de 10.53 mm; do derivado clorofenol 9.9 mm e do hipoclorito de sódio 6.2 mm. O digluconato de clorexidine e o derivado cloro-fenol apresentaram redução significante no crescimento dos dermatófitos testados, enquanto o hipoclorito de sódio, na concentração recomendada, demonstrou baixa atividade antifúngica contra os dermatófitos testados.

First isolation of the Stephanoascus ciferrii in feline otitis in Brazil

Ear infections in cats are uncommon, especially involving yeasts. This report describes the first isolation of the Stephanoascus ciferrii, teleomorph of the Candida genus, in a case of feline otitis in Brazil. The identification and characterization of Stephanoascus ciferrii were confirmed by the Vitek2 System (BioMerieux ®).

Aflatoxicose em cães na região Sul do Rio Grande do Sul / Aflatoxicosis in dogs in Southern Rio Grande do Sul

Descrevem-se os aspectos clinicopatológicos de casos de aflatoxicose em cães no Sul do Rio Grande do Sul. Foi realizado um estudo retrospectivo dos casos diagnosticados como aflatoxicose em cães necropsiados no Laboratório Regional de Diagnóstico (LRD) da Faculdade de Veterinária da Universidade Federal de Pelotas (UFPel) no período de 1978 a 2012. Em quatro casos o diagnóstico foi confirmado pela detecção de níveis de 89 a 191 ppb de aflatoxinas B1 e G1 no alimento dos cães. De um total de 27 cães com cirrose hepática, em seis havia suspeita de aflatoxicose pelas lesões macro e microscópicas e pelo tipo de alimentação que os cães recebiam. Os sinais clínicos nos casos confirmados e nos suspeitos caracterizaram-se por apatia, diarreia, icterícia e ascite, com evolução para morte em 8 a 30 dias nos casos confirmados e em 15 a 60 dias nos casos suspeitos. A dieta era à base de derivados de milho ou arroz, farelo de amendoim e, em um caso suspeito, a dieta era ração comercial. As alterações macroscópicas caracterizaram-se por ascite, icterícia, fígado aumentado de tamanho, com ou sem nódulos, hemorragia nas serosas, conteúdo intestinal hemorrágico. Os casos foram classificados de acordo com o padrão histológico principal, caracterizado por vacuolização difusa no citoplasma de hepatócitos nos casos agudos, por proliferação de ductos biliares e discreta fibroplasia nos casos subagudos e por fibrose acentuada nos casos crônicos. Aparentemente, a enfermidade não é importante como causa de morte em cães na região, no entanto, alerta-se para a possibilidade de casos com diagnóstico de cirrose hepática sem causa determinada serem causados por aflatoxicose.

Clinical pathological aflatoxicosis in dogs is described in southern Rio Grande do Sul. It was conducted a retrospective study of cases diagnosed as aflatoxicosis in dogs necropsied at the Regional Diagnostic Laboratory (LRD) of the Veterinary School of the Federal University of Pelotas (UFPel) in the period 1978-2012. In four cases the diagnosis was confirmed by detection of levels of aflatoxins B1 and G1, with the finding of 89-191ppb in the feed. The macroscopic and histologic lesions and the diet observed in six of 27 dogs with liver cirrhosis led to suspicion of aflatoxicosis. Clinical signs evidenced in confirmed or suspected cases were lethargy, diarrhea, jaundice and ascites, progressing to death within 8 to 30 days in confirmed cases, and within 15 to 60 days in suspected cases. The diet was corn and rice byproducts and peanut meal, and one of the dogs received commercial ration. Gross changes were characterized by ascites, jaundice, enlarged liver, with or without regenerative nodules, hemorrhages in serous membranes and bloody intestinal content. The cases were classified according to the main histological pattern, characterized by diffuse vacuolation of the cytoplasm of hepatocytes in acute cases, by proliferation of bile ducts, and mild fibrosis in subacute cases, and by severe fibrosis in chronic cases. Apparently the disease is not important as a cause of death in dogs in the region, nevertheless the possibility of cases of cirrhosis of unknown etiology would be caused by aflatoxicosis.

Microsporose mista canina - relato e caso / Microporosis mixta canina ? relato de caso / Canine mcrosporose mixed - case report

Dermatofitoses são micoses causadas por fungos como: Epidermophyton, Microsporum e Trichophyton. M. canis e M. gypseum são os principais responsáveis pelas infecções dermatofíticas em pequenos animais e acometem principalmente animais jovens. Essas micoses cursam com lesões nas camadas queratinizadas da pele e pelos e em infecções mistas, a intensidade das lesões pode ser aumentada, pois os agentes causais atuam potencializando o quadro clínico. Descreve-se um caso de microsporose mista canina em um filhote de cão da raça Rottweiler. Um canino da raça Rottweiller atendido em uma clínica particular, pois apresentava lesões alopécicas e descamativas em diferentes sítios do corpo. Foi realizada coleta por meio de raspado cutâneo e o material foi encaminhado para exame micológico. O exame direto do pelo revelou estruturas características de dermatofitose com hifas artrosporadas e aglomerados de artroconídeos. O cultivo micológico resultou em crescimento de colônias fúngicas sugestivas de M. gypseum e M. canis nos meios de cultura semeados com confirmação das características macro e micromorfológicas das duas diferentes espécies fúngicas. Assim, mediante processamento micológico estabeleceu-se, juntamente com o histórico do caso, o diagnóstico de microsporose mista canina. Desse modo, conclui-se a necessidade de exames laboratoriais para a confirmação do diagnóstico definitivo de dermatofitose, sendo a profilaxia, prevenção, controle e potencial zoonótico dessa micose dependente da espécie infectante, tendo importância o seu controle também por seu impacto em saúde pública.

Dermatophytosis mycoses are caused by fungi such as Epidermophyton, Microsporum and Trichophyton. M. canis and M. gypseum are the main responsible for dermatophytic infections in small animals and affect mainly young animals. These mycoses occur with lesions in the keratinized layers of the skin and the mixed infections and the intensity of the lesions can be increased because the causative agents act enhancing the clinical picture. We describe a case of mixed microsporose dog in a puppy dog breed Rottweiler. A canine breed Rottweiller served in a private clinic because he had alopecia, scaly lesions at different body sites. It was collected through skin scrapings and the material was sent for mycological examination. Direct examination of the structures revealed by the characteristics of dermatophytosis with of hyphae artrosporadas and arthroconidia clusters. The mycological cultivation resulted in growth of fungal colonies suggestive of M. canis and M. gypseum in culture media seeded with confirmation of macro and micromorphological characteristics of the two different fungal species. Thus, by processing mycological established together with the case history, the diagnosis of canine microsporose mixed. Through the described case is concluded the need for laboratory tests to confirm the definitive diagnosis of dermatophytosis and prophylaxis, prevention, control and zoonotic potential of this mycosis dependent on the infecting species, taking control of the same importance also for their impact on public health.

Dermatofitosis son micosis causadas por hongos como: Epidermophyton, Microsporum y Trichophyton. M.canis y M. gypseum que son los principales responsables en las infecciones por dermatofitos en animales pequeños y afectan principalmente animales jóvenes. Esas micosis se producen con lesiones en las capas queratinizadas de la piel, pelos y en infecciones mixtas, la intensidad de las lesiones puede ser aumentada, pues los agentes causantes actúan potencializando el cuadro clínico. Se describe un caso de microsporosis mixta canina en un cachorro de perro de la raza Rottweiller. Un canino de la raza Rottweiller atendido en una clínica privada que presentaba lesiones de alopecia y lesiones escamosas en diferentes partes del cuerpo. Se realizó recolección por medio de raspado cutáneo y el material fue enviado para examen micológico. El examen directo del pelo reveló estructuras características de dermatofitosis hifas artrosporadas y aglomerados de artroconidios. El cultivo micológico resultó en crecimiento de colonias fúngicas sugestivas de M. gypseum y M canis en los medios de cultura sembrados, con confirmación de características macro y micro morfológicas de dos especies fúngicas. Así, mediante procesamiento micológico establecido junto con el histórico del caso, el diagnóstico de microsporosis mixta canina. Se concluye la necesidad de exámenes de laboratorio para confirmación del diagnóstico definitivo de dermatofitosis, siendo que la profilaxis, prevención, control y potencial zoonótico de esa micosis depende de la especie infectante, siendo importante su control por el impacto que causa a la salud pública.

Isolation of dermatophytes in wild felids from screening centers

The aim of this study was detect the presence of dermatophyte fungi on wild felids from screening centers. Samples were taken from 30 animals, assembled in two groups: "free-ranging" and "transitory captivity". The dermatophytes (Trichophyton genus), isolated from two felids (6.6%), both of the group "free-ranging".

Pesquisa de trematódeos digenéticos em Heleobia spp. (Mollusca: Hydrobiidae) em área de ocorrência da Ehrlichiose monocítica equina, no Rio Grande do Sul, Brasil / Research of trematodes digenetics from Heleobia spp. (Mollusca: Hydrobiidae) in area of occurrence of equine monocytic ehrlichiosis, in Rio Grande do Sul, Brazil

A ehrlichiose monocítica equina (EME) na região Sul do Rio Grande do Sul tem demonstrado ser importante nas criações de cavalos Crioulos. A enfermidade foi relatada e diagnosticada como causa de diarreia, prejuízos com tratamentos e com a morte de equinos não estabulados, sendo apontada como um fator limitante na criação de equinos em algumas regiões. O modo de transmissão pela via oral, intermediada por trematódeos em ambientes aquáticos, tem sido sustentado. Caracóis dulciaquícolas estão envolvidos como hospedeiros intermediários de trematódeos albergadores de Neorickettsia risticii. Um total de 16.846 caracóis Heleobia foi coletado nos municípios de Arroio Grande, Rio Grande, Palmares do Sul e Santa Vitória do Palmar, 92,2% dos quais foram encontrados nas raízes de aguapés (Eichornea spp.). A frequência de trematódeos presentes nos caracóis variou de 2,3 a 12,8% nas propriedades coletadas. Foram encontrados três tipos de cercárias, morfotipo 1, morfotipo 2 e morfotipo 3, e dois morfotipos de metacercárias nos caracóis. Um total de 357 insetos da ordem Odonata foi coletado, fases de metacercárias foram encontradas no tegumento da subordem Anisoptera com frequência de 5,3%. Mais estudos são necessários para identificar as fases larvais encontradas, bem como para conhecer o hospedeiro definitivo, identificar o parasito adulto e a relação de seu ciclo de vida com a ocorrência da ehrlichiose monocítica equina.

The equine monocytic ehrlichiosis in the region South of the Rio Grande do Sul has demonstrated to be important in the creations of Crioulo horses. It has been reported as cause of diarrhea in equine not surround and as cause of losses with treatments and death of the animals, being pointed as a limited factor in the range breeding in some regions. The way of transmission for the oral way, intermediated for trematodes in aquatic environments, has been supported. Freshwater snails are involved as intermediate hosts of trematodes and harbor of the Neorickettsia risticii. A total of 16,846 Heleobia snails had been collected in the cities of Arroio Grande, Rio Grande, Palmares do Sul and Santa Vitoria do Palmar, 92.2% of which had been found in the roots of aquatics plants (Eichornea spp.). The frequency of trematodes present in the snails varied of 2.3 to 12.8% in the collected regions. Three types of cercariae were found, morphology type 1, morphology type 2 and morphology type 3, and two morphologic type of metacercariae of the snails. A total of 357 insects of the Odonata order were collected, stages of metacercariae had been found in the tissues of suborder Anisoptera with 5.3% of frequency. More studies are necessary for identification of the joined larval phases, as well as knowing the host definitive and identifying the adult parasite and the relation of its cycle of life with the occcurrence of equine monocytic ehrlichiosis.

Clinical and mycological analysis of dog's oral cavity

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.

Efeito antifúngico de agentes químicos sobre leveduras com potencial patogênico isoladas de ambiente hospitalar veterinário / Antifungal effect of chemical agents in yeasts with pathogenic potential isolated from veterinary hospital environment

Os ambientes hospitalares podem albergar micro-organismos patogênicos e oportunistas, capazes de infectar, particu- larmente, os indivíduos imunossuprimidos. O processo de limpeza e desinfecção assume importância capital para o controle de tais doenças. O presente trabalho avaliou a suscetibilidade de leveduras isoladas de superfícies de ambiente hospitalar veterinário frente a quatro desinfetantes/antissépticos comumente utilizados na rotina de desinfecção. O teste de microdiluição em caldo foi realizado com 24 isolados leveduriformes frente ao hipoclorito de sódio 4%, cloreto de benzalcônico 2%, clorexidina-cetrimida 6,6% e derivado de cloro-fenol 3%. A clorexidina-cetrimida, cloreto de ben- zalcônio e derivado de cloro-fenol foram eficazes em todos os isolados com concentração inibitória mínima e concen- tração fungicida inferiores à concentração recomendada pelo fabricante. Por outro lado, a ação fungicida do hipoclorito de sódio em 79,1% dos isolados testados foi obtida na concentração recomendada pelo fabricante, com desempenho inferior aos demais desinfetantes avaliados.

Hospital environments can have pathogens and opportunistic pathogens, important for immunocompromising individuals. The process of cleaning and disinfection constitutes an important measure for the control of these diseases. This study evaluated the sensitivity of yeasts isolated from surfaces of nosocomial veterinary environment to four disinfectants/antiseptics used in the routine of disinfection. The test of broth microdilution was carried in 24isolates of yeasts against 4% sodium hypochlorite, 2% benzalkonium chloride, 6.6% chlorexidine-cetrimide and 3% chloro-phenol derivate. Chlorexidine-cetrimide, benzalkonium chloride and chloro-phenol derivate were efficient in all isolates with minimal inhibitory concentration and minimal fungicide concentration lower than the concentra- tion recommended by manufacturer. By other hand, fungicide action of the sodium hypochlorite in the concentration recommended by the manufacturer was verified in 79.1% of the isolated tested, with the lower performance of than the others evaluated disinfectants.

Alterações hematológicas em felinos com esporotricose cutânea / Hematological alterations of cats with cutaneous sporotrichosis / Alterações hematológicas en felinos con esporotricosis cutânea

A esporotricose é a micose subcutânea de maior ocorrência em pequenos animais, especialmente felinos e caninos. Esta doença causa lesões cutâneas ulceradas de caráter subagudo ou crônico que podem resultar no comprometimento do estado geral do animal. Este estudo avaliou alterações hematológicas em gatos com esporotricose relacionando-as com a forma clínica da doença. As alterações hematológicas foram observadas em 73,3% dos animais, sendo caracterizadas principalmente por leucocitose por neutrofilia e anemia. Os resultados demonstram que animais com a forma cutânea fixa da micose não apresentam alterações no hemograma, enquanto animais com a forma cutânea disseminada desenvolvem importantes alterações que indicam o agravamento do quadro clínico com possibilidade de evolução da micose para uma forma sistêmica.

Sporotrichosis is the most frequent subcutaneous mycosis of small animals, specially cats and dogs. This disease leads toof the mycosis do not present alterations in the blood count. However, animals with the cutaneous disseminated form develop important alterations that can aggravate the disease and evolve the mycosis to a systemic form.

La esporotricosis es la micosis subcutánea de mayor ocurrencia en pequeños animales, específicamente felinos y caninos. Esta enfermedad produce lesiones cutáneas ulceradas de carácter sub agudo o crónico que pueden resultar en el comprometimiento del estado general del animal. Este estudio evalúo alteraciones hematológicas en gatos con esporotricosis, relacionándolas con la forma clínica de la enfermedad. Se ha observado alteraciones hematológicas en 73,3% de los animales, siendo caracterizadas principalmente por leucocitosis por neutrofilia y anemia. Los resultados demuestran que animales con la forma cutánea fija de la micosis no presentan alteraciones en el hemograma, mientras que los animales con la forma cutánea diseminada desarrollan importantes alteraciones que indican el comprometimiento del cuadro clínico con posibilidad de evolución de la micosis para una forma sistémica.

Sporothrix schenckii in a hospital and home environment in the city of Pelotas/RS - Brazil

This study describes the isolation of S. schenckii in hospital and home environments in Brazil. Samples were collected from surfaces of a veterinary service place and at home. S. schenckii was detected in 1.5 percent of the samples from the hospital environment. However, this fungus was isolated from all sampled areas in home environments. The isolation of S. schenckii deonstrates that these surfaces could act as infection sources to anials and huans. Therefore, employees and pet owners could be exposed to this agent, and the contamination, through surfaces, could occur through the traumatic inoculation of the fungus or by direct contact with pre-existing lesions.

Esse estudo descreve o isolamento de S. schenckii em ambiente hospitalar e domiciliar, no Brasil. Foram colhidas amostras de superfície de local de atendimento veterinário e ambiente domiciliar. S. schenckii foi isolado em 1,5 por cento das amostras do ambiente hospitalar. Entretanto, esse fungo foi isolado em todas as amostras do ambiente domiciliar. O isolamento do S. schenckii demonstra a importância dessas superfícies atuarem como fontes de infecção para animais e humanos. Portanto, funcionários e proprietários de animais de estimação estariam expostos a esse agente e a contaminação, através das superfícies, poderia ocorrer pela inoculação traumática do fungo ou pelo contato direto com lesões pré-existentes.

Caracterização da microbiota por fungos filamentosos no tegumento hígido de bovinos de corte / Characterization of filamentous fungal flora from the integument of healthy cattle

O tegumento de bovinos pode albergar uma grande diversidade de fungos filamentosos, potencialmente capazes de causar dermatoses, incluindo fungos do grupo dos dermatófitos. O presente estudo teve por objetivos caracterizar a microbiota de fungos filamentosos do tegumento hígido de bovinos de corte, verificar a presença de dermatófitos como parte integrante da microbiota, assim como avaliar a ocorrência de dermatófitos no solo dos estabelecimentos estudados. Durante o período de um ano, 56 bovinos sadios de quatro propriedades da fronteira Oeste do RS foram submetidos a coletas mensais de pelos e raspado cutâneo. No mesmo período, foram obtidas amostras de solo para pesquisa de dermatófitos. As análises dos resultados indicaram uma microbiota de fungos filamentosos classificada em 30 gêneros, dos quais os mais frequentes foram (nesta ordem): Nigrospora, Fusarium, Curvularia, Alternaria, Epicoccum, Paecilomyces e Trichoderma. Nas amostras de pelame, obteve-se o isolamento de Trichophyton mentagrophytes (0,4 por cento), Microsporum gypseum (0,3 por cento) e Trichophyton verrucosum (0,1 por cento). M. gypseum foi o dermatófito mais frequentemente isolado do solo. Os resultados obtidos evidenciaram que a microbiota fúngica do tegumento hígido de bovinos de corte comporta-se de maneira transitória e que bovinos sadios não atuam como portadores assintomáticos de T. verrucosum, sugerindo que esta espécie de dermatófito é não residente no tegumento dos animais.

The tegument of cattle may harbor a great diversity of filamentous fungi that can potentially cause skin diseases, including fungi belonging to the dermatophyte group. This study aimed to characterize the population of filamentous fungi from a healthy coat of cattle, checking for dermatophytes as part of the microbiota, as well as evaluating the presence of dermatophytes in the soil occupied by the animals. During the period of one year, 56 healthy cattle in four properties in the western boundary of the state of Rio Grande do Sul, Southern Brazil, were subjected to monthly samples of hair and skin scrapings. In the same period soil samples were obtained for the detection of dermatophytes. The results indicated a fungal biota classified into 30 different genera, among which the most frequent were (in order of frequency): Nigrospora, Fusarium, Curvularia, Alternaria, Epicoccum, Paecilomyces and Trichoderma. Isolates of Trichophyton mentagrophytes (0.4 percent), Microsporun gypseum (0.3 percent), and T. verrucosum (0.1 percent) were obtained in the samples of skin scrapings. M. gypseum was the dermatophyte most frequently isolated from soil. The results showed that the fungal biota of the external tegument of healthy cattle is transient. Moreover, it was shown that healthy cattle do not act as asymptomatic carriers of T. verrucosum, suggesting that this species of dermatophytes is not resident in the tegument of animals.

Evaluación de la actividad bactericida de aceites esenciales de hojas de guayabo, pitango y arazá / Evaluation of the bactericidal action of essential oils from guava, Surinam cherry and strawberry guava

Introducción: guayabo, pitango y arazá son árboles fructíferos, nativos de América del Sur. El té de sus hojas es utilizado popularmente en el tratamiento de la diarrea. Objetivo: evaluar la actividad antimicrobiana de los aceites de las hojas Psidium guajava L., Eugenia uniflora L. y Psidium cattleyanum Sabine y sus constituyentes. Métodos: los aceites esenciales fueron extraídos con un aparato tipo Clevenger modificado. Su análisis se realizó en cromatógrafo (GC/MS). Se evaluó la actividad antimicrobiana de los aceites obtenidos frente a Escherichia coli, Salmonella typhimurium y Staphylococcus aureus, mediante la técnica de microdilución en caldo con determinación de la concentración bactericida mínima del aceite. Resultados: el aceite del guayabo presentó actividad bactericida frente a E. coli en la concentración de 2 y 8 por ciento frente al Staphylococcus aureus y Salmonella typhimurium. El aceite de arazá no presentó actividad bactericida frente a los microorganismos testados y el aceite de pitango presentó actividad bactericida en la concentración de 8 por ciento frente al Staphylococcus aureus y Salmonella typhimurium. Los constituyentes principales encontrados en común en los aceites de las 3 plantas fueron el a-copaeno y a-humuleno. Conclusiones: los aceites presentaron actividad bactericida frente a los microorganismos testados y los constituyentes en ellos encontrados son compatibles con los encontrados en otros trabajos.

Introduction: Guava, Surinam cherry and strawberry guava are native South American fruit trees and the tea from their leaves is traditionally used to treat diarrhea. Objective: to evaluate the antimicrobial action of Psidium guajava L., Eugenia uniflora L. and Psidium cattleyanum Sabine and their constituents. Methods: the essential oils were extracted by a modified Clevenger. The analysis of essential oils was performed on chromatograph (GC/MS). The antimicrobial activity of obtained oils against Escherichia coli, Salmonella typhimurium and Staphylococcus aureus was evaluated by broth microdilution assay to determine their minimum bactericidal concentration. Results: the guava oil showed bactericidal activity against E. coli at 2 percent and 8 percent concentration rates against S. aureus and S. typhimurium. The strawberry guava oil showed no bactericidal activity against tested microorganisms whereas Surinam cherry had bactericidal activity at 8 percent concentration against S. aureus and S. typhimurium. The main constituents commonly found in the three plant oils were a-copaene and a-humulene. Conclusions: the oils showed bactericidal activity against tested microorganisms and the constituents are comparable to those found in other studies.