Het vóórkomen en de rol van Clostridium difficile bij gezonde pups, en pups met acute diarree.

Abstract

Puppies are regularly presented to the veterinary practice with symptoms of acute diarrhea. In addition to non-infectious causes, infectious diseases are also a common cause of this phenomenon. One of the possible pathogens of these symptoms could be the gram-positive, anaerobic bacterium Clostridium difficile. C. difficile can produce 3 types of toxins: enterotoxin TcdA, cytotoxin TcdB, and binary toxin CDT.28, 32 Since the turn of the century there seems to be an increase in the number of human cases with a C. difficile infection.9, 32.43 In veterinary medicine this bacterium also seems to play an increasingly important role.3, 21,43 To assess the incidenceand the possible pathogenic role of C. difficile in puppies with diarrhea and healthy puppies, a case control study was launched. This study was performed with a case group which consisted of the feces of 46 puppies with acute infectious diarrhea, and a control group which contained the feces of 24 puppies without diarrhea. These fecal samples were tested for the presence of C. difficile and other infectious causes as worm, protozoa, viral agents and other bacteria (i.e. E.coli, Campylobacter spp.). The fecal samples were provided by veterinary practices throughout the Netherlands. Each sample was sent to the research facility together with a pair of surveys completed by veterinarian and owner. Through these questionnaires, risk factors were determined for the occurrence of diarrhea symptoms in combination with the presence of C. difficile. The fecal samples were tested for the presence of C. difficile, by means of a direct culture as well as an indirect culture 7-days enriched broth before culture on selective agarplates) were used. Positive samples were confirmed by a Glu-D PCR and sent to the Leiden University Medical Centre for ribotyping and determination of the presence of toxin genes. 14/70 (20%) of the samples were positive for C. difficile, of which 9/46 (19.6%) in the case group and 5/24 (20.8%) in the control group. The most frequently encountered ribotype was non-toxinogenic type 010. Both in the control group and in the case group, a potentially pathogenic variant was found; Ribotype 012 in the case group (1x) and ribotype 045 in the control group (1x). In this study, there appears to be no significant difference in the occurrence of C. difficile in puppies with acute infectious diarrhea and puppies without diarrhea. The surveys were used to examine or antibiotic use, sex, owners having multiple pets, contact in the past 2 weeks with animals with diarrhea or vomiting symptoms, and visiting places where multiple dogs congregate, could be considered as a risk factor for the fecal excretion of C. difficile. This was not the case. The outcome of this study showed that C. difficile has no role in puppies with acute diarrhea