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Therefore, further studies are necessary to identify reservoirs or potential sources of virulent STEC strains in order to establish control and prevention strategies for STEC associated diseases in animals and humans. We demonstrated the presence of the STEC virulence genes in healthy and diarrhoeic Austrian calves but the importance of the virulence factors of STEC (stx1, stx2, eae and Ehly) in calf diarrhoea and systemic disease is not well defined. The detection rates of other enteropathogens were 25.7% bovine coronavirus, 11.7% Cryptosporidium spp., 10.4% Eimeria spp., 9.1% group A rotavirus and Giardia spp. were present in 1.3% and 0.4% of all samples. Clostridium perfringens was detected in twenty-one samples, the most prevalent toxin type of Clostridium perfringens was found to be type A (76.2%). eaeA and Ehly genes were detected more frequently in the strains from diarrhoeic calves 57.1% and 50.0%, respectively. coli positive samples observed, only two carried the Shiga toxin genes: stx1, in a diarrhoeic calf and both stx1 and stx2 in a healthy calf. Overall, 35 out of the 230 (15.2%) samples analyzed carried the Shiga toxin gene: stx1, stx2 or both stx1 and stx2 in their faeces, STEC.
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coli F5 was identified only in one calf without diarrhoea. Escherichia coli was detected in 17% of all the faecal samples and was more prevalent in healthy calves. Our data suggested that DNA microarray with its high efficiency and accuracy could be used as an alternative to the culture method.įaecal samples from 230 diarrhoeic and healthy calves aged 0-6 weeks, from 100 farms in Austria, were examined between October 2004 and February 2005 for the presence of bacteria, especially Shiga toxin-producing Escherichia coli (STEC), viruses and parasites. Using stool culture as a control, gene-chip sensitivity was 100%, specificity 95.2%, and index of accurate diagnosis 0.952. Of the 1,500 clinical cases, 32.7% of the patient stools were positive for bacteria.
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The limit of detection was approximately 10(3) CFU/ml for one species of pathogen. Our data showed that the probes of the assay were successful in discriminating 14 genera or species of intestinal pathogens. Basic information from patients was collected and analyzed. To determine the consistency of DNA microarray and culture method, 1,500 samples of clinical diarrheal stool and 200 samples of normal stool from healthy individuals were examined in a double-blind fashion.
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Hybridization between probes and amplicons was performed. Primers and 21 oligonucleotide probes based on sequences of the bacterial 16SrRNA gene were arrayed on microarray slides. To establish and evaluate a quick and accurate DNA microarray method to detect intestinal pathogens directly from human diarrheal stool samples as an alternative to traditional culture methods. Treatment with rehydration solutions and provision of dry and warm conditions are vital in the treatment of calf diarrhoea. Accurate and rapid early confirmation of the etiology in the disease outbreak as well as improving the various management factors are advised, for effective control and prevention of enteric disease in newborn calves. Various laboratory methods have been applied for the detection of infectious agents of calf diarrhea in fecal sample such as, bacterial culture, electron microscopy, molecular based techniques (PCR, DNA microarray) and serological techniques (enzyme-linked immunosorbent assay, latex agglutination test). The most prominent virulence factors identified in bacterial diarrhea are expression of fimbrial (pili) antigens that enables the bacteria to adhere and to colonize the luminal surface of the small bowel and elaboration of one or more enterotoxins that influence intestinal secretion of fluids. Clostridium perfringens and Campylobacter species have also been identified as causes of enteric diseases in calf diarrhea other, Non-infectious factors, such as insufficient uptake of colostrum, poor sanitation, stress, overcrowding in the calf pens and cold weather, could cause neonatal calf diarrhea. Among the bacterial causes of diarrhea in neonatal food animals, Escherichia coli and Salmonella species are the most common and economically important ones. Calf diarrhea is a multi factorial disease entity that can have serious financial and animal welfare implications in both dairy and beef sucker herds and is one of the most common diseases reported in calves up to 3 months old. Infectious agents and the calf itself are the major constraints for raising replacement stock. Calf diarrheic diseases result from complex interactions of the environment.