The aim of the study was to present cases of botulism in animals found in Poland in 2019–2021. The analytical laboratory diagnosis and difficulties that occurred in the interpretation of the results are described. From 2019 to 2021, samples of serum, intestinal content, liver, spleen, kidney, faeces, wet feed, dry feed, ensilage, water and mixed samples of internal organs associated with 10 suspected animal botulism cases were sent to the National Veterinary Research Institute. Samples were analysed using a mouse bioassay and culture methods in combination with ntnh and bont gene detection. Among the ten putative botulism cases, only four (40%) were confirmed in the laboratory on the basis of the detection of botulinum toxin (BoNT) or the ntnh or bont genes. The remaining six (60%) were determined as probable despite observable characteristic clinical signs. The diagnosis of botulism in animals is a very difficult task, made so by the heterogeneity of Clostridium botulinum strains and possible loss of toxinogenicity during laboratory processing or the potential degradation of toxins. Laboratory diagnosis is a complex and problematic process which should utilise different prescribed methods for specific types of sample.

Many neurotoxigenic clostridia are found in soil. Among animals, birds are especially susceptible to botulism, perhaps because they feed on insects, invertebrate carcasses, and decayed feeds contaminated with spores of Clostridium (C.) botulinum. C. botulinum type C is mainly involved in avian botulism. In the summer of 2005, death of a mute swan (cygnus olor) living in the pond of large bird cage was found in Seoul Grand Park Zoo. The birds presented presumptive clinical signs of botulism, such as ruffled hackle feathers, abnormal posture of the head, weakness, and flaccid paralysis. At that time, pond water in the breeding facilities was drained for 7 days, but there were still remained water containing sediment of feed and feces. Therefore, botulism was suspected and an experimentation were made to detect C. botulinum in the dead mute swan. Gross post-mortem findings of a mute swan showed jelly-like hemorrhagic contents in the intestine, sands and vegetations in the stomach. C. botulinum was isolated from the liver, small intestine and large intestine samples. Botulism was also confirmed by mouse inoculation test with the organ samples. With PCR, a gene encoding C. botulinum type C toxin was detected for the several organs of the mute swan died. These results suggested that death of mute swan was caused by C. botulinum type C.