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The characterisation of Bacillus spores occurring in the manufacturing of (low acid) canned products

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Author: Oomes, S.J.C.M. · Zuijlen, A.C.M. van · Hehenkamp, J.O. · Witsenboer, H. · Vossen, J.M.B.M. van der · Brul, S.
Institution: TNO Kwaliteit van Leven
Source:International Journal of Food Microbiology, 1-2, 120, 85-94
Identifier: 240314
doi: doi:10.1016/j.ijfoodmicro.2007.06.013
Keywords: Biology · Biotechnology · Cluster analysis · Food processing · Molecular phylogenetic characterisation · Spore heat resistance · acid · metal ion · RNA 16S · acidity · article · Bacillus subtilis · bacterial spore · bacterium contamination · canned food · DNA DNA hybridization · fatty acid analysis · food contamination · food industry · food preservation · food processing · gene sequence · Geobacillus stearothermophilus · heat tolerance · molecular typing · nonhuman · phylogeny · ribotyping · RNA gene · sporogenesis · strain identification · survival · validation process · Bacillus · Bacterial Typing Techniques · Cluster Analysis · Colony Count, Microbial · Consumer Product Safety · Fatty Acids · Food Contamination · Food Handling · Food Microbiology · Food Preservation · Germination · Heat · Humans · Phylogeny · RNA, Ribosomal, 16S · Species Specificity · Spores, Bacterial · Bacillus (bacterium) · Bacteria (microorganisms)


Spore-forming bacteria can be a problem in the food industry, especially in the canning industry. Spores present in ingredients or present in the processing environment severely challenge the preservation process since their thermal resistance may be very high. We therefore asked the question which bacterial spore formers are found in a typical soup manufacturing plant, where they originate from and what the thermal resistance of their spores is. To answer these questions molecular techniques for bacterial species and strain identification were used as well as a protocol for the assessment of spore heat stress resistance based on the Kooiman method. The data indicate the existence and physiological cause of the high thermal resistance of spores of many of the occurring species. In particular it shows that ingredients used in soup manufacturing are a rich source of high thermal resistant spores and that sporulation in the presence of ingredients rich in divalent metal ions exerts a strong influence on spore heat resistance. It was also indicated that Bacillus spores may well be able to germinate and resporulate during manufacturing i.e. through growth and sporulation in line. Both these spores and those originating from the ingredients were able to survive certain thermal processing settings. Species identity was confirmed using fatty acid analysis, 16SrRNA gene sequencing and DNA-DNA hybridisation. Finally, molecular typing experiments using Ribotyping and AFLP® analysis show that strains within the various Bacillus species can be clustered according to the thermal resistance properties of their spores. AFLP® performed slightly better than Ribotyping. The data proofed to be useful for the generation of strain specific probes. Protocols to validate these probes in routine identification and innovation aimed at tailor made heat processing in soup manufacturing have been formulated. © 2007 Elsevier B.V. All rights reserved.