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Food Spoilage: “Stinkies”, “Slimies” and Biofilms -byJovana Kovačević October 22, 2009 Outline Spoilage of Food o Spoilage Signs o Meat, Fish, Dairy o What do laboratory tests tell you about spoilage? Biofilms o Properties and formation o Why are they a problem? How do we deal with spoilage and biofilms? 10/22/2009 2 Food Spoilage Undesirable deterioration of food quality that may result in changes in odor, taste and appearance of food. 10/22/2009 3 Food Spoilage Types of spoilage: Microbial: Bacteria, yeasts, molds Non-microbial Foreign materials, enzymes Changes not necessarily harmful 10/22/2009 4 Microbial Food Spoilage Each food unique microbial environment unique spoilage agents for each Why do microorganisms spoil our food? food composition ideal source of nutrients for microbes availability of water and oxygen suitable pH Factors that affect spoilage: Oxygen, temperature, pH, aw, humidity 10/22/2009 5 Spoilage organisms Similar microflora emerge in different foods under the same conditions Affected by the: o Type of food o Processing o Preservation o Storage conditions Domination of specific spoilage microbes: 10/22/2009 o Initial counts present o Nutrient composition o Chemical and physical parameters 6 Spoilage Signs Odor: Breakdown of proteins – putrefaction e.g. “rotten egg” smell Sliminess Discoloration 10/22/2009 Mold on bread, blue and green mold on citrus fruit and cheese 7 Spoilage Signs… Souring Production of acid e.g. sour milk from production of lactic acid Gas formation Meat becomes spongy Swollen or bubbling packages and cans 10/22/2009 8 Food Types: Based on rate of spoilage 1) Highly perishable meat fruit milk vegetables eggs WET 2) Semi perishable potatoes nuts 3) Stable 10/22/2009 rice flour dry beans DRY 9 Common Causes of Food Spoilage Inadequate storage temperatures Prolonged storage times Improper ventilation Cross contamination Excessive delays between receiving and storing Inadequate food safety standards 10/22/2009 10 Meat Spoilage 10/22/2009 11 Meat spoilage… Cutting board contamination Conveyor belts Temperature Delay between storage and distribution Fecal contamination from intestines 10/22/2009 12 Meat Spoilage Continued Food Types of Spoilage Spoilage Microorganisms MEAT Putrefaction Clostridium, Pseudomonas, Proteus, Alcaligenes, Chromobacterium Souring Chromobacterium, Lactobacillus, Pseudomonas Mouldy Penicillium, Aspergillus, Rhizopus Souring Pseudomonas, Micrococcus, Bacillus Greening Lactobacillus spp., Streptococcus, Pediococcus Slimy Leuconostoc Souring Greening Lactobacillus, Carnobacterium, Leuconostoc Odor, Slime Pseudomonas, Alcaligenes, Xanthomonas Fresh Cured Vacuum Packed Poultry 10/22/2009 13 Fish and Eggs Spoilage 10/22/2009 16 Fish and Eggs Spoilage Continued Fish Polluted waters Transportation vehicles, boxes Poultry and Eggs Human contact Penetration by bacteria Cracks 10/22/2009 TEMPERATURE PHYSICAL DAMAGE INTRINSIC FACTORS 17 Fish & Eggs Spoilage Continued Food FISH EGGS 10/22/2009 Types of Spoilage Spoilage Microorganisms Discoloration Pseudomonas Putrefaction Chromobacterium, Halobacterium, Micrococcus Green rot Pseudomonas Colorless rot Pseudomonas, Alcaligenes, Chromobacterium Black rot Coliforms Fungal rot Proteus, Penicillium, Mucor 18 Spoilage of Milk and Dairy Products 10/22/2009 http://farm3.static.flickr.com/2313/2181694147_12ce5cd763.jpg 19 Spoilage of Milk and Dairy Products Continued Food Types of Spoilage Spoilage Microorganisms Bitterness Pseudomonas spp. Souring Lactobacillus thermophilus Sweet curdling Bacillus cereus Green discoloration Penicillium Green to black discoloration Cladosporium Black discoloration Candida Sliminess (high pH) Pseudomonas spp. “Gassy” cheese Coliforms, LAB, Clostridia DAIRY MILK (pasteurized) CHEESE 10/22/2009 20 Spoilage of Fruits and Vegetables 10/22/2009 http://timelytidbits.wordpress.com/2008/07/28/the-word-of-god-as-rotting-fruit/ 21 Spoilage of Fruits and Vegetables Food FRESH FRUITS AND VEGETABLES 10/22/2009 Types of Spoilage Spoilage Microorganisms Bacterial soft rot Erwinia carotovera, Pseudomonas spp. Gray mould rot Botryitis cinerea Rhizopus soft rot Rhizopus nigrican Blue mould rot Penicillium italicum Black mould rot Aspergillus niger, Alternaria Sliminess and Souring Saprophytic bacteria 22 Examples of Food Quality Tests Meat & Fish: pH for vacuum packaging (< 6.0) Visual inspection for leakage / color change Faulty seals, pin-holes Sensory test Aerobic plate count at 30°C Indication of hygiene at packaging Expected refrigerated shelf-life ↑ APC = improper cleaning of equipment, unacceptable time/temperature history of raw products 10/22/2009 23 Testing for Microbial Quality Total Viable Count Enterobacteriacea Coliforms Escherichia coli 10/22/2009 Enterococci Staphilococcus aureus Bacillus spp. Clostridium perfringens Lactic acid bacteria Pseudomonas species Yeasts and molds 24 10/22/2009 25 Examples of Food Submission Forms 10/22/2009 26 Total Viable Count Total Viable Count also known as: Heterotrophic plate count (HPC) Aerobic plate count (APC) Total plate count (TPC) Aerobic colony count (ACC) Aerobic mesophilic count Purpose: The aerobic plate count gives the total number of bacteria able to grow in an oxygenated or aerobic environment. Cooked foods should have little to no bacteria present Result provides a guide for shelf-life of foods The higher the count: prolonged storage, and/or inadequate cooking. Note: Not a useful indicator for fermented foods or fresh ready-to-eat raw fruits and vegetables. 10/22/2009 27 Enterobacteriaceae A family of Gram-negative, facultatively anaerobic, rod-shaped bacteria that do not form endospores and ferment variety of sugars Includes: Pathogenic microbes: e.g. pathogenic E. coli, Proteus, Salmonella, Shigella, Yersinia, Enterobacter, Serratia, Klebsiella Non-pathogenic microbes: e.g. Citrobacter, Erwinia, Hafnia 10/22/2009 28 Coliforms and E. coli Coliforms: Total coliforms Fecal coliforms: o Found in human and animal intestinal wastes o More precise indicator of the presence of sewage contamination than total coliforms Four groups of fecal coliforms: o Enterobacter o Klebsiella o Escherichia o Citrobacter 10/22/2009 29 Escherichia coli Best indicator of fecal sanitation problems Characterized by the fermentation of glucose and lactose Found In the gut of warm blooded mammals In the gut of insects and reptiles As natural soil microbial flora 10/22/2009 30 Sanitary Quality Guidelines Food Group Indicator Test Result (CFU/g) Satisfactory Unsatisfactory <5 >5 < 100 > 1000 < 100,000 > 1,000,000 Long shelf life fish products, meat products, fruit and vegetable products < 1,000,000 > 100,000,000 Fermented foods & Fresh fruits and vegetables Not applicable Not applicable <100 > 1000 <3 >3 <3 >3 Aerobic Colony Count Canned foods & Cooked hot-held food Cooked chilled food no handling Cooked chilled food with handling & Preserved foods Total Coliform Count All foods except fresh fruits & vegetables, or foods containing them Fecal Coliform Count All foods except fresh fruits & vegetables, or foods containing them Escherichia coli Count All foods Aerobic Plate Count: Environmental Swabs Guidelines for interpretation of bacterial counts from swabs and sponges as means of monitoring the cleanliness of surfaces. Interpretation Clean Contaminated Very Contaminated CFU Count Per Area Swabbed Log Conversion < 45 CFU < 1.65 140 to 260 CFU 2.15 – 2.41 > 260 CFU > 2.41 Counts on Surface* (based on 25 cm2 surface area sampled) < 5 CFU / cm2 ~ 5 to 10 CFU / cm2 > 10 CFU / cm2 ** Values will vary depending on size of the surface area sampled. To calculate the number of bacteria per cm2 take the total CFU and divide by the surface area swabbed. 10/22/2009 32 Common Spoilage Microorganisms Shewanella putrefaciens Pseudomonas spp. o Gram negative o Rod shaped o Motile o Aerobic o Non-spore forming o Biofilm formers 10/22/2009 http://www.lille.inra.fr/lille_eng/unites_et_recherches/nos_unites/ gpta/interfaces_bacteries_aliments_surfaces_solides 33 Pseudomonas aeruginosa http://www.microbelibrary.org/asmonly/details.asp?id=545&Lang= Biofilms “Variety of microorganisms arranged in a complex relationship to one another and embedded in a mass of extracellular polysaccharides of their own making” 10/22/2009 (http://www.personal.psu.edu/faculty/j/e/jel5/biofilms/) 35 Biofilms Continued Communities of bacteria adhering to: http://www.microbelibrary.org/ Environmental surfaces Living tissues Indwelling medical devices Industrial or potable water system piping Natural aquatic systems Food processing facilities: o Floors, waste water pipes, bends in pipes, rubber seals, conveyor belts, stainless steel surfaces (Kumar and Anand, 1998) FEMS Microbiology Letters. 228:203-210. Why do bacteria form biofilms? Protection from the environment o Sanitizers o Antibiotics Nutrient availability and metabolic cooperativity Acquisition of new genetic traits 10/22/2009 38 Biofilm formation 10/22/2009 http://www.personal.psu.edu/faculty/j/e/jel5/biofilms/ 39 Control of Microbial Spoilage Good manufacturing practices Cleaning and sanitation Processing environment Equipment Handling with minimal physical damage Washing (in some case) Proper storage temperature Rapid movement of food through processing plant Treatments… preservation 10/22/2009 41 Food preservation Temperature based: o o o o o Chilling Refrigeration Freezing Pasteurization Canning Water activity reduction o Dehydration o Addition of salts and sugars Irradiation Modified Atmosphere Packaging Fermentation Treatment with chemicals o Sorbic acid, phenylphenates, diphenyl and iodophors, fumigation with sulfur-containing dusts 10/22/2009 42 References 1. Davey and O’Toole (2000). Microbial Biofilms: from ecology to molecular genetics. Microbiol. Molec. Bio. Rev. 64(4):847-867. 2. Forsythe, S.J, and P.R. Hayes. 1998. Food Hygiene Microbiology and HACCP. 3rd Ed. pp. 434. Aspen Publishers Inc., Maryland. 3. ICMSF. 2005. Microorganisms in Foods. Microbial Ecology of Food Commodities. 2nd Ed. pp. 763. Kluwer Academic/Plenum Publishers, New York, NY. 4. Marsh, E.J., H. Luo, H. Wang. 2003. A three-tiered approach to differentiate Listeria monocytogenes biofilm-forming abilities. FEMS Microbiology Letters. 228:203-210. 5. http://www.personal.psu.edu/faculty/j/e/jel5/biofilms/ 6. http://www3.sympatico.ca/tania.nicolas/River%20Biofilm.jpg 7. http://www.lille.inra.fr/lille_eng/unites_et_recherches/nos_unites/ gpta/interfaces_bacteries_aliments_surfaces_solides 8. http://www.personal.psu.edu/faculty/j/e/jel5/biofilms/ 9. http://www.microbelibrary.org/asmonly/details.asp?id=545&Lang= 10. http://timelytidbits.wordpress.com/2008/07/28/the-word-of-god-as-rotting-fruit/ 11. http://farm3.static.flickr.com/2313/2181694147_12ce5cd763.jpg 10/22/2009 43 Thank you! 10/22/2009 44