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Tuija Pirttijärvi Contaminant Aerobic Sporeforming Bacteria In View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Helsingin yliopiston digitaalinen arkisto Tuija Pirttijärvi Contaminant Aerobic Sporeforming Bacteria in the Manufacturing Processes of Food Packaging Board and Food Department of Applied Chemistry and Microbiology Division of Microbiology University of Helsinki Academic Dissertation in Microbiology To be presented, with the permission of the Faculty of Agriculture and Forestry of the University of Helsinki, for public criticism in auditorium 1041, at Viikki Biocenter, Viikinkaari 5, on July 21st, 2000, at 12 o’clock noon. Helsingin yliopiston verkkojulkaisut Helsinki 2000 ISBN 952-91-2304-3 (pdf) Supervisor: Prof. dr. Mirja Salkinoja-Salonen Academy Professor Department of Applied Chemistry and Microbiology University of Helsinki Helsinki, Finland Reviewers: Prof. dr. Tuula Honkanen-Buzalski Department of Food Microbiology National Veterinary and Food Research Institute Helsinki, Finland Prof. dr. Seppo Salminen Department of Biochemistry and Food Chemistry University of Turku Turku, Finland Opponent: dr. Anders Christiansson Swedish Dairy Association Research and Development Lund, Sweden To Reijo, Anja and Jutta CONTENTS List of original papers .................................................................................................................1 Abbreviations..............................................................................................................................2 Abstract.......................................................................................................................................3 1. Introduction.............................................................................................................................4 2. Review of the literature...........................................................................................................5 2.1. History and taxonomy of aerobic sporeforming bacteria .................................................5 2.2. Occurrence of bacilli in foods and in food processes.......................................................5 2.3. The significance of bacterial contamination in paper and board production....................9 2.4. Degradation of starch by bacteria ....................................................................................9 2.5. Control of contaminant bacteria in paper industry.........................................................11 2.6. Nisin and its applications ...............................................................................................12 3. Aims of the study ..................................................................................................................14 4. Materials and Methods..........................................................................................................14 5. Results and discussion ..........................................................................................................14 5.1. Bacilli contaminating paper and board manufacturing process and their properties related to food spoilage and food safety ...............................................................................14 5.2. Detection and typing of B. cereus in industrial processes..............................................20 5.3. Identifying the emetic toxin producing strains of B. cereus...........................................22 5.4. Preservation of surface sizing starch against deterioration by bacilli.............................24 6. Conclusions...........................................................................................................................27 7. Tiivistelmä ............................................................................................................................28 8. Acknowledgements...............................................................................................................30 9. References.............................................................................................................................32 Original publications List of original papers I Pirttijärvi, T.S.M., Graeffe, T.H. and Salkinoja-Salonen, M.S. 1996. Bacterial contaminants in liquid packaging boards: assessment of potential for food spoilage. Journal of Applied Bacteriology 81, 445-458. II Pirttijärvi, T.S.M., Ahonen, L.M., Maunuksela, L.M. and Salkinoja-Salonen, M.S. 1998. Bacillus cereus in a whey process. International Journal of Food Microbiology 44, 31-41. III Pirttijärvi, T.S.M., Andersson, M.A., Scoging, A.C. and Salkinoja-Salonen, M.S. 1999. Evaluation of methods for recognising strains of the Bacillus cereus group with food poisoning potential among industrial and environmental contaminants. Systematic and Applied Microbiology 22, 133-144. IV Pirttijärvi, T.S.M, Wahlström, G., Rainey, F.A., Saris, P.E.J. and Salkinoja-Salonen, M.S. 200x. Inhibition of bacilli in industrial starches by nisin. Submitted to Journal of Industrial Microbiology and Biotechnology. The author’s contribution Paper I: Tuija Pirttijärvi wrote the paper, is the corresponding author and interpreted the results. She was responsible for all experimental work except for the mass fragmentographic confirmation of the fatty acids and the partial 16S rDNA sequencing. Paper II: Tuija Pirttijärvi wrote the paper, is the corresponding author and interpreted the results. Paper III: Tuija Pirttijärvi wrote the paper, is the corresponding author and interpreted the results. She did all experimental work except for the determination of emetic toxin, a part of the enterotoxin tests, transmission electron microscopy and the serotyping. Paper IV: Tuija Pirttijärvi wrote the paper, is the corresponding author and interpreted the results. She did all experimental work except for nisin activity analysis and the partial 16S rDNA sequencing. 1 Abbreviations AOAC Association of Official Analytical Chemists ATCC American Type Culture Collection ATP adenosine triphosphate cfu colony forming unit BCET-RPLA Bacillus cereus enterotoxin reversed passive latex agglutination BHI brain heart infusion BUGM+G Biolog universal growth medium with 1 % glucose CDase cyclodextrinase CGTase cyclodextrin glycosyl transferase D140 decimal reduction time at 140 °C DBNPA 2,2-dibromo-3-nitrilopropionamide DSM(DSMZ) Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH EDTA ethylenediaminetetraacetic acid EELA National Veterinary and Food Research Institute ELISA-VIA Bacillus diarrhoeal enterotoxin visual immunoassay FDA Food and Drug Administration HACCP hazard analysis critical control point IDF International Dairy Federation ISO International Organization for Standardization IU international unit kb kilo basepairs m threshold below which all results are considered satisfactory (in microbiological safety criteria of foods) M acceptability threshold, above which results are not considered satisfactory (in microbiological safety criteria of foods) Mb mega basepairs MIC minimum inhibitory concentration MUF methyl umbelliferyl NCFB National Collection of Food Bacteria NMKL The Nordic Committee on Food Analysis O.D. optical density PCA plate count agar PDA potato dextrose agar PFGE pulsed field gel electrophoresis PEG polyethylene glycol SMR Swedish Dairy Association TAPPI Technical Association of the Pulp and Paper Industry TE Tris-EDTA TSA tryptone soy agar TSB tryptone soy broth TSBA tryptone soy broth agar (BBL) UPGMA unweighted pair group method with arithmetic mean 2 Abstract The significance of bacilli in industrial processes was studied, with focus on food grade paper and paperboard production and on a dairy process. Food packaging paper and board contained as contaminants mainly sporeforming bacteria belonging to the genera Bacillus, Paenibacillus and Brevibacillus. The contaminants were usually found in quantities from <50 to 250 cfu g-1 homogenized paperboard, which are lower than in many foods. Of those frequently found, especially the Bacillus cereus group, B. licheniformis, B. subtilis, and Brevibacillus brevis are species known to be capable of hydrolysing food components, and some strains of these species produce food poisoning toxins or may grow at refrigerated temperatures. The diversity among the contaminant B. cereus isolates in the whey processing line of a cheese dairy was large. The whey line of the cheese dairy was colonized by specific communities of phenotypically non-conventional types of B. cereus. Great genetic diversity was found among contaminant B. cereus strains from paper industry and from different dairies. We found that the phenotypic properties on which the standard methods (e.g. ISO, FDA, IDF) rely for recognising B. cereus (lecithinase activity, nitrate reduction), did not match with the properties of B. cereus isolates found as contaminants in industrial processes. Industrial isolates, including the toxigenic ones, often missed one or several of the typical B. cereus characters even in cases where 100 % 16S rDNA identity was found with B. cereus or with B. thuringiensis. The whole cell fatty acid compositions of a group of industrial contaminant B. cereus isolates deviated so much from those in the widely used commercial database that the strains were only poorly recognised or not recognised at all as B. cereus. Better identification was obtained with signature fatty acids 11-methyldodecanoic acid and trans-9-hexadecenoic acid, which were present without exception in all of the more than 200 industrial contaminant isolates representing
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