Botulism is a paralytic disease caused by the botulinum neurotoxin produced by Clostridium botulinum. In the summer season, intensive outbreaks of avian botulism were reported in both poultry and wild birds, including 5 Korean native chicken farms, 1 pheasant farm, and 1 community of spot-billed duc...
Botulism is a paralytic disease caused by the botulinum neurotoxin produced by Clostridium botulinum. In the summer season, intensive outbreaks of avian botulism were reported in both poultry and wild birds, including 5 Korean native chicken farms, 1 pheasant farm, and 1 community of spot-billed ducks. The affected domestic birds showed 24.5% to 58.3% mortality with specific clinical signs including ataxia, limber neck and diarrhea. To confirm the botulinum toxin, neutralization tests were performed on sera (4 Korean native chicken farms and 1 pheasant farm) or culture supernatant (spot-billed ducks). Additionally, the contents of the cecum and liver from poultry presenting signs suggestive of botulism were inoculated to isolate the pathogen. The toxin genes were then detected by polymerase chain reaction (PCR). Through the neutralization tests, it was possible to diagnose the botulism and, except in the case of one Korean native chicken farm, to identify the type of pathogen. Using detection by PCR, except in two cases of the Korean native chicken farms, the botulinum toxin gene was found. Additionally, in four cases, it was possible to identify the C/D mosaic type using PCR. A new diagnostic method for detection of avian botulism using single tube nested PCR. The purpose for developing this method was to overcome disadvantages of mouse bioassay. Three primer pairs including antisense primer were designed targeting the N terminal of toxin gene of Clostridium botulinum type C and C/D. The limit of detection was determined with purified DNA and spiking spores, and its result were 1.1 pg/μℓ and 4.3 spores/200 mg, respectively. The specificity of the PCR assay was evaluated on 27 reference bacterial strains and on samples of caecum from botulism outbreak farms. In addition, the PCR assay results on cecum contents of 300 healthy chickens (150 Korean native chickens and 150 broilers) were all negative. This method could be rapid and easy compared to mouse bioassay, evading ethical problem. It is also economic in compared with real time PCR.
Botulism is a paralytic disease caused by the botulinum neurotoxin produced by Clostridium botulinum. In the summer season, intensive outbreaks of avian botulism were reported in both poultry and wild birds, including 5 Korean native chicken farms, 1 pheasant farm, and 1 community of spot-billed ducks. The affected domestic birds showed 24.5% to 58.3% mortality with specific clinical signs including ataxia, limber neck and diarrhea. To confirm the botulinum toxin, neutralization tests were performed on sera (4 Korean native chicken farms and 1 pheasant farm) or culture supernatant (spot-billed ducks). Additionally, the contents of the cecum and liver from poultry presenting signs suggestive of botulism were inoculated to isolate the pathogen. The toxin genes were then detected by polymerase chain reaction (PCR). Through the neutralization tests, it was possible to diagnose the botulism and, except in the case of one Korean native chicken farm, to identify the type of pathogen. Using detection by PCR, except in two cases of the Korean native chicken farms, the botulinum toxin gene was found. Additionally, in four cases, it was possible to identify the C/D mosaic type using PCR. A new diagnostic method for detection of avian botulism using single tube nested PCR. The purpose for developing this method was to overcome disadvantages of mouse bioassay. Three primer pairs including antisense primer were designed targeting the N terminal of toxin gene of Clostridium botulinum type C and C/D. The limit of detection was determined with purified DNA and spiking spores, and its result were 1.1 pg/μℓ and 4.3 spores/200 mg, respectively. The specificity of the PCR assay was evaluated on 27 reference bacterial strains and on samples of caecum from botulism outbreak farms. In addition, the PCR assay results on cecum contents of 300 healthy chickens (150 Korean native chickens and 150 broilers) were all negative. This method could be rapid and easy compared to mouse bioassay, evading ethical problem. It is also economic in compared with real time PCR.
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