Sarcocystis spp. detection in cattle using different diagnostic methods / Detecção de Sarcocystis spp. em bovinos usando diferentes métodos de diagnóstico

Cattle are considered intermediate hosts of Sarcocystis, which can cause clinical signs and lower performance in the acute phase of infection. Sarcocystis spp. are usually not visible to the naked eye during the post mortem inspection. Moreover, fresh microscopic examination and transmission electron microscopy techniques are difficult to apply to large samples. Therefore, extensive studies on Sarcocystis infection in cattle using molecular and serological methods are required. Here, we investigated Sarcocystis spp. infection in cattle using fresh microscopic examination and polymerase chain reaction of myocardium samples and compared the results with the presence of antibodies against Sarcocystis spp. in corresponding serum samples detected using indirect fluorescent antibody test. Microscopic Sarcocystis were observed in 100% of the myocardial samples, and Sarcocystis DNA was present in 86% (43/50) of these samples. Antibodies against Sarcocystis spp. were detected in 96% (48/50) and 80% (40/50) of the serum samples at 1:25 and 1:200 dilutions, respectively. The three associated methods (fresh microscopic examination, PCR and serology) showed good sensitivity and detection for Sarcocystis spp. compared with fresh microscopic examination (only), and they may facilitate diagnosis in live animals on a large scale as well as monitoring of the herd status.

Os bovinos são considerados hospedeiros intermediários de Sarcocystis, podendo causar sinais clínicos e menor desempenho na fase aguda da infecção. Sarcocystis spp. geralmente não são visíveis a olho nu durante a inspeção post mortem. Além disso, o exame microscópico a fresco e as técnicas de microscopia eletrônica de transmissão são difíceis de aplicar a uma amostras de grande tamanho. Portanto, são necessários extensos estudos sobre a infecção por Sarcocystis em bovinos usando métodos moleculares e sorológicos. Aqui, investigamos a infecção de Sarcocystis spp. em bovinos por meio de exame microscópico a fresco e reação em cadeia da polimerase de amostras de miocárdio e comparado os resultados com a presença de anticorpos contra Sarcocystis spp. em amostras de soro correspondentes detectadas usando o teste de anticorpos fluorescentes indiretos. Sarcocistos microscópicos foram observados em 100% das amostras de miocárdio, e o DNA de Sarcocystis estava presente em 86% (43/50) dessas amostras. Anticorpos contra Sarcocystis spp. foram detectados em 96% (48/50) e 80% (40/50) das amostras de soro nas diluições 1:25 e 1:200, respectivamente. Os três métodos associados (exame microscópico a fresco, PCR e sorologia) mostraram boa sensibilidade e detecção para Sarcocystis spp. em comparação com o exame microscópico fresco (apenas) e podem facilitar o diagnóstico em animais vivos em larga escala, bem como o monitoramento do status do rebanho.

Efficacy of aqueous garlic extract on growth, aflatoxin B1 production, and Cyto-morphological aberrations of Aspergillus flavus, causing human ophthalmic infection: topical treatment of A. flavus keratitis

By using agar well diffusion assay, antifungal activity of aqueous extract prepared from Egyptian garlic (Allium sativum L.) was evaluated in vitro against two strains of Aspergillus flavus (OC1 and OC10) causing human ocular infection. The recorded minimum inhibitory concentration (MIC) for growth inhibition of both strains was 3.60 mg/ml. Aqueous garlic extract (AGE) was used in successive in vivo tests as an attempt to cure rabbit's fungal keratitis caused by A. flavus OC1. Findings showed that diluted preparation of AGE was effective topical antifungal agent and succeeded to cure severe A. flavus keratitis in a time course less than 10 days without any observable side effects. Microscopic examination showed that AGE induced deleterious cyto-morphological aberrations inA. flavus target cells. AGE applied to Czapek's broth via contact method was more effective on growth, spores and aflatoxin B1 production than AGE applied to the same broth at the same concentration via fumigation method.(AU)

Use of direct microscopic examination for Paracoccidioides brasiliensis diagnosis at Instituto Adolfo Lutz (IAL) Regional Laboratories Network, state of São Paulo, Brazil / Exame direto no diagnóstico de Paracoccidioides brasiliensis realizados nos Laboratórios Regionais do Instituto Adolfo Lutz, São Paulo, Brasil

Paracoccidioidomycosis (PCM),an autochthonous disease from Latin America,is a systemic disease caused by a dimorfic fungus Paracoccidioides brasiliensis. It is supposed that this microorganism infects hosts when conidia in aerosol form are inhaled by individuals, causing primary acute, subacute,or even chronic lung infection. In order to give evidence of the significance in diagnosing PCM by means of direct microscopic examination, which is of low cost, and easy to perform assay, a study was carried out analyzing clinical samples from IAL Regional Laboratories Network collected during the period from January 1996 to December 2002. A total of 3,807 clinical samples from lung, and 25 extra-pulmonary clinical samples were prepared by conventional procedures. Significant quantity of exam request forms with incomplete data on patient age, profession,and birthplace was observed. Additionally, samples from male patients were more frequently requested for PCM diagnosis than from female subjects. Positive lung samples were detected in 4%, being predominantly from male patient aged from 31 to 70 years old. Positive extra-lung samples were also detected, which demonstrated the presence of this disease in other organs. Although PCM mortality rate have still been increasing in Brazil, due to systemic mycosis, PCM has not included among diseases for compulsory notification yet. PCM has still

Paracoccidioidomicose (PCM) é uma doença sistêmica causada pelo fungo dimórfico Paracoccidioides brasiliensis. Apresenta-se como infecção pulmonar primária assintomática, aguda, sub-aguda ou crônica. Para ratificar a importância do exame direto, de baixo custo e complexidade, no diagnóstico da PCM, foi realizado um levantamento de amostras clínicas nos Laboratórios Regionais do IAL (LR-IAL) e Laboratório I Araçatuba - DIR VI, no período de janeiro de 1996 a dezembro de 2002. Foram analisadas 3807 amostras de origem pulmonar e 25 amostras extrapulmonares, preparadas de acordo com suas peculiaridades. Detectou-se significativa quantidade de solicitações de exames com informações incompletas quanto aos dados do paciente. Foi demonstrada positividade em 4% das amostras pulmonares (predominando no adulto do sexo masculino) assim como em amostras extrapulmonares (88%). A diferença na demanda natural das solicitações para este diagnóstico, entre os Regionais, pode estar relacionada com endemicidade da região, busca e/ou suspeita da doença e encaminhamento das amostras para os Laboratórios de Referência da Rede Pública. Após 98 anos da descrição pioneira por Adolfo Lutz, a doença ainda é foco de preocupação para a Saúde Pública.