Application of PCR for detection of Clostridium botulinum type D in bovine samples

Diagnosis of botulism in cows is obtained by detecting the neurotoxin and/or Clostridium botulinum in the suspected animal. The standard method for detecting the toxin is the mouse bioassay. However, in recent years, the use of mice has become very costly and inconvenient in some facilities, and public pressure has been increasing to find alternatives to live animal bioassays. In this manuscript, we describe the use of the polymerase chain reaction (PCR) procedures in the diagnosis field cases of bovine type D botulism. Bovine samples from clinical cases diagnosed as C. botulinum type D according by clinical symptoms and bioassay resulted in expected PCR product ( approximately 497 bp) similar to the C. botulinum type D NCTC 8265 strain while the gene product was confirmed by sequence data.