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

Abstract

The aim of the study was the analysis of Lithuania n honeys for the presence of the spores of Clostridium botulinum . The analyses were performed on multifloral, honey dew, nectar-honeydew, rape, buckwheat, heather and linde n 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) an d Tripticase Peptone Glucose Yeast (TPGY) enrichment broths. Bacteria of typical phenotypic features were obtained by culture method from 48 honey samples. Identifica tion of Clostridium botulinum types A, B, E and F was performed with the use of multipl ex-PCR method. The prevalence analysis showed 30 (60%) samples contaminated with Clostrdium botulinum spores. The major serotype detected was type A: 20 (66.6%) posi tive 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 typ e of C. botulinum spores was observed in 3 honey samples. Keywords: Botulism, Contamination , C. botulinum spores, Cooked meat medium, Multiplex-PCR, TPGY.