Prevalence of Clostridium botulinum Type A, B, E and F Isolated From Directly Sold Honey in Lithuania

The aim of the study was the analysis of Lithuanian honeys for the presence of the spores of Clostridium botulinum. The analyses were performed on multifloral, honeydew, nectar-honeydew, rape, buckwheat, heather and linden honey bought in direct sale. Prior to culture on solid media the samples were prepared by supernatant filtration method and cultured in parallel in Cooked Meat Medium (CMM) and Tripticase Peptone Glucose Yeast (TPGY) enrichment broths. Bacteria of typical phenotypic features were obtained by culture method from 48 honey samples. Identification of Clostridium botulinum types A, B, E and F was performed with the use of multiplex-PCR method. The prevalence analysis showed 30 (60%) samples contaminated with Clostrdium botulinum spores. The major serotype detected was type A: 20 (66.6%) positive honey samples; while type B was found in 7 samples (23.3%), type E in 4 samples (13.3%) and type F in 3 (10%) positive honey samples. Contamination with more than one type of C. botulinum spores was observed in 3 honey samples.