Abstract
The Clostridium difficile is a Gram-positive opportunist, anaerobic, spore-forming rod found in the soil, water and enteric microbiota of many animal species. It has been described as the cause for enteritis in human beings and animals. In swine it has grown in importance due to the large number of cases of neonatal enteritis that affects the colon of one to seven-day-old piglets. This bacterium produces two kinds of toxins: A (enterotoxin) and B (cytotoxin). They have an important role in the disease’s pathogenesis. Aiming at researching the presence of the bacterium in up to seven days piglets submitted to antibiotic therapy as well as the production of the toxins A and B in isolated samples, 8 collections were made in different regions of the state of Santa Catarina, totalizing 490 samples of stool and retal swabs of piglets, gathered in the period of January to March 2008. The retal swabs were processed in the same day they were collected in the Microbiology Lab of CAV/UDESC and the stools were frozen at -4ºC in steril Ependorf so that they could be submitted to the ELISA test later. Twenty-tree colonies of C. difficile were isolated, but no one produced the A and B toxins according to the ELISA test. Of the 69 stools analyzed, 32 (46.37%) were positive samples, 3 (4.34%) were intermediate samples, and 34 (49.27%) were negative samples, according to the ELISA test.KEYWORDS: Antibiotic, Clostridium difficile, diarrhea, swine.
Highlights
The Clostridium difficile is a Gram-positive opportunist, anaerobic, spore-forming rod found in the soil, water and enteric microbiota of many animal species
Clostridium difficile é uma bactéria Grampositiva, anaeróbia, formadora de esporos, encontrada no meio ambiente e no trato intestinal de vários mamíferos, pássaros e répteis (NAGY & BILKEI, 2003)
Das 23 amostras isoladas, nenhuma produziu as toxinas, pela análise do teste de ELISA
Summary
Para realização do presente estudo as amostras do material foram obtidas de maternidades de suínos localizadas nas seguintes regiões de Santa Catarina: Vale do Itajaí (Rio do Sul e Ituporanga), Meio-Oeste Catarinense (Concórdia, Jaborá, Luzerna e Erval Velho) e região Sul (Braço do Norte e São Ludgero), no período de janeiro a março de 2008. Os suabes retais foram processados no mesmo dia das coletas, e as fezes foram congeladas a – 20 oC para posterior realização do teste de ELISA para identificação das toxinas A e B. Os suabes retais foram semeados pela técnica de esgotamento neste meio e incubados anaerobicamente por 48 horas a 37 oC em jarras com Gapak, que proporcionou uma atmosfera com os seguintes gases: N2 (80%), CO2 (10%) e H2 (10%). O teste de ELISA foi utilizado para determinar qualitativamente a produção das toxinas A e B pelo Clostridium difficile nas amostras de fezes e nas colônias isoladas. Empregaram-se as primeiras 46 unidades do kit ELISA para correlacionar a produção das toxinas pelas colônias isoladas com a produção das toxinas das suas respectivas fezes. As amostras cujo valor de absorbância fica mais de 10% acima do CUT-OFF calculado foram consideradas positivas e as amostras que ficam mais de 10% abaixo do CUTOFF calculado foram consideradas negativas, sempre seguindo as instruções do fabricante
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