Food Microbiology: Good, Bad & Ugly

Microbial association on red smear cheese
• Cheese types: Münster, Romadour, Limburger, Harzer, Vacherin Mont d`Or, Tilsiter, Livarot,and many others • Bacteria:Arthrobacter nicotianum, Brevibacterium linens, Corynebacterium ammoniagenes, C. casei, C. variabile, Microbacterium gubbeenense,Rhodococcus fascians, Staphylococcus equorum, S. saprophyticus, and many others. • Yeasts: Debaryomyces hansenii, Kluyveromyces marxianus, Pichia membranaefaciens • Fungi: Galactomyces geotrichum


Staphylococci involved in food fermentation
S. xylosus: fermented sausages, raw ham, read smear cheese (strains forming enterotoxin E were involved in human infections) fish sauce fish sauce fish sauce, ham, fermented sausages horse skin, red smear cheese, fermented sausages, curing brines, raw ham red smear cheese had been identified in French cultures

S. piscifermentans: S. condimenti: S. carnosus: S. equorum:

S. succinus ssp casei S. warneri S. saprophyticus

The safe tradition in food fermentation of strains of certain Staphylococcus species confirms that within a taxonomically related group of organisms safe strains or even species exist. The same applies e.g. to enterococci, streptococci (S. thermophilus), Aspergillus oryzae ( used for koji production and closely related to A. flavus).


Probiotics - definitions
Published Definition Substances produced by microorganisms which promote the growth of other microorganisms Organisms and substances which contribute to intestinal microbial balance A live microbial feed supplement which beneficially affects the host animal by improving its intestinal microbial balance A viable mono- or mixed-culture of microorganisms which applied to animal or man, beneficially affects the host by improving the properties of the indigenous microcflora Living microorganisms, which upon ingestion in certain numbers, exert health benefits beyond inherent basic nutrition Reference Lilly and Stillwell 1965 Parker 1974 Fuller 1989 Havenaar and Huis In’t Veld 1992 Schaafsma 1996 Naidu et al. 1999

A microbial dietary adjuvant that beneficially affects the host physiology by modulating mucosal and systemic immunity, as well as improving nutritional and microbial balance in the intestinal tract

Live microorganisms which when administered in adequate amounts confer a health benefit on the host.

A preparation of or a product containing viable, defined microorganisms in sufficient numbers, which alter the microflora (by implantation or colonization) in a compartment of the host and by that exert beneficial health effects in this host Live microorganisms which when administered in adequate amounts confer a health benefit on the host.

Schrezenmeir and de Vrese 2001 FAO/WHO report, October 2001

Probiotics – requirements
• Probiotics must be alive. • Probiotics must be safe. • Probiotics must deliver a measured physiological benefit, substantiated by studies conducted in the target host. • Probiotics needn’t be restricted to food applications or oral delivery. • Probiotics used as pharmaceuticals or as topical agents are not excluded from this definition. • A definition of probiotics shouldn’t limit the mechanism of action. • Survival of gastrointestinal tract transit or impact on normal flora shouldn’t be required.
Sanders 2003


such as: • • • • • • Gastro-intestinal probiotics Skin probiotics Scalp probiotics Probiotics of the oral cavity Probiotics of the underarm and feet Urogenital (including vaginal) probiotics 4 .Expected Benefits Associated with Consumption of Probiotics •Increased tolerance to infections •Control of diarrhea •Reduction of blood pressure •Cholesterol reduction •Allergy control •Immunomodulation •Cancer reduction Human probiotics Human probiotics can be grouped according to the ecological niches of the human body they occupy (or the part of the body they deliver their health-promoting action to).

acidilacticic Sporolactobacillus inulinusa Strep. reuteri L. plantarum L. faecium Lactoc. adolescentis B. longum other LAB Ent. infantis B. rhamnosus Bifidobacteria B. thermophilus non-LAB Bacillus cereus (‘toyoi‘)a. E. lactisc Leuconostoc mesenteroidesc Ped. Metchnikoff 5 . gasseri L. johnsonii L. faecalisa Ent. breve B. d Escherichia coli (‘Nissle 1917‘)d Propionibacterium freudenreichiia. bifidum B. lactisb B. animalis B.Microorganisms in probiotic products Lactobacilli L. paracasei L. acidophilus (L. crispatus L. casei) L. d Saccharomyces cerevisiae (‘boulardii‘)d The Bulgarian farmer's longevity and healthy life are the result of the consumption of fermented dairy products. gallinaruma L. http://www. ..If the science of probiotics holds up in clinical trials.yogurt could replace many pharmaceutical drugs.usprobiotics.htm Fermented Vegetables •sauerkraut •olives •cucumbers* •carrots •celery •beans •peas •corn •tomatoes *Controlled fermentation •peppers •onions •citron •beets •turnips •radishes •chard •Brussels sprouts •cauliflower 6 .

leafy undergrowth CHIP DIP: If you can take it out of its container and bounce it on the the mayonnaise is spoiled FROZEN FOODS: Frozen foods that have become an integral part of the defrosting problem in your freezer compartment will probably be spoiled -. the meat is spoiled BREAD: Sesame seeds and Poppy seeds are the only officially acceptable "spots" that should be seen on the surface of any loaf of bread. the egg is probably past its prime MAYONNAISE: If it makes you violently ill after you eat smells normal and tastes normal: you eat enough to make you ill from the ingested pathogens or toxins • Spoiled food does not normally cause food poisoning because it is rejected by the consumer before ingestion Food Spoilage Test • • • • • EGGS: When something starts pecking its way out of the shell. or dense. branches.Food Spoilage Microorganisms • It is important to be able to distinguish food poisoning from food spoilage • Food poisoning is when food is eaten which looks normal. Fuzzy and hairy looking white or green growth areas are a good indication your bread has turned into a pharmaceutical laboratory experiment FLOUR: Flour is spoiled when it wiggles CANNED GOODS: Any canned goods that have become the size or shape of a softball should be disposed of POTATOES: Fresh potatoes do not have roots.htm • • • • 7 .ntu. it has gone bad http://science.(or wrecked anyway) by the time you pry them out with a kitchen knife MEAT: If opening the refrigerator door causes stray animals from a threeblock radius to congregate outside your house.

Food Spoilage Test GENERAL RULE OF THUMB • Most food cannot be kept longer than the average life span of a •Release of enzymes .slower •extracellular •intracellular (lysis) 8 .htm Microbial Food Spoilage •Microbial growth in a food .uk/external/foodmicrobiol. http://science. • Keep a goldfish nearby your refrigerator to gauge this.

atmosphere • “Proper” abuse of temperature • Sufficient time of storage Microbial food spoilage: “first come – first serve” •Bacteria •Yeast •Molds 9 .Microbial food spoilage: favorable conditions • Transfer of the microorganisms into the food • Optimal environment: • AW. nutrients.

Caused by preformed toxin in the food. Clostridium botulinum and Staphylococcus aureus • Food infection . H2 or H2S2 • Slime formation: due to the production of dextrans and/or amount of microorganisms Foodborne diseases • Food poisoning . organism may or may not be alive and growing. organism multiply once food is ingested.Microbial Food Spoilage Changes in Food Quality • Odor: due to production of volatile endcompounds • Color: pigment production or oxidation • Texture: softening due to the breakdown of pectin in vegetables or the tissues by proteinases • Accumulation of gas: CO2. Salmonella 10 .Live cells delivered by contaminated food.

500 950 2. 1999 The incidence of diagnosed foodborne infections is a small fraction of total 11 .000 2.000 10..500 2.000 110.500 Deaths 560 100 26 500 124 375 Mead et al.Public health burden of major foodborne infections Pathogen Salmonella Campylobacter Shiga-toxin E.350.500 Hospitalizations 16.000 112.300 20. EID 5:707-25.000 2. coli Listeria Norovirus Toxoplasma Cases 1.

F Staphylococcus aureus FOOD POISONING SYNDROME • • • • onset: recovery: major symptoms: other symptoms: 0. B. non-motile. D.5 to 6 hours 24 to 72 hrs vomiting. diarrhea nausea. asporogenous cocci • “grape-like” clusters • enterotoxin • effective at 1ug/kg • protein of 239 amino acids • serological types: A. C. prostration 12 . cramps. retching.Staphylococcus aureus • Gr+. salivation.

average 2. aureus from inflamed lesion on the hand of the cook who had made the breakfast.5 h • symptoms: diarrhea (88%). then heated. • Breakfast prepared day before flight.5 h after breakfast. 1 of 20 crew • food eaten: 1 h post Anchorage. • Held at room temperature during the flight. • Isolated SED from omelet and ham. held at room temperature for 6 h during preparation • Held 14. crew 5.Staphylococcus aureus case study • Flight from Tokyo to Copenhagen via Anchorage • 196 of 343 passengers.6 h post Anchorage. 13 . aureus from fecal swabs of 5 patients and from left over omelet and ham. served to all 5. cheese omelet w/ ham passengers only • onset: 0. steak dinner.5 to 5. nausea (68%) Staphylococcus aureus investigation • Isolated SED-producing S. vomiting (82%) cramps (74%). • Isolated SED-producing S.5 h post Anchorage.5 h at 10°C prior to the flight. snack.

3 ug) will kill a 104 kg person.1ng • Dose of 3. few cases but high mortality (25%). • Very heat sensitive 14 . destroyed by 10 min in 80 oC • Flacid paralysis of muscles (acetylcholine release is blocked) • Common in soil and water • Improper canning -> spore germination -> toxin production -> canned food used without cooking -> disease • Infant botulism: consumption of honey that is contaminated by spores (0 .Botulism (C. botulinum) • The most potent toxin known. • Cultures easily produce 105 to 106 mld per ml.000 mld (0.2 months) • Treatment: antitoxin and ventilation Clostridium botulinum toxin • 1 mld = 0.

and ate it two and one-half days later without reheating. weakness. in August). Mother left pot pie out on shelf (in California. 15 . botulinum) Botulism and Temperature Abused Pot Pie (Morbidity and Mortality Weekly Report.Botulism (C. Vol. • Illness: next day. difficulty breathing. # 3. respiratory arrest. Type A botulinum toxin was isolated from the left-over food and patient's serum. Father brought home take-out hamburgers. chest pain. January 1983). • Patient: 56 year old diabetic woman • Symptoms: diplopia. • Scenario: Son prepared pot pie for mother. 32.

2) forms larger complexes. 3) channel formation.C. 14 different toxins and genes in species. cpe action . perfringens Product Turkey Chicken Pork Ground pork Beef Ground beef Fish Shrimp Overall % positive 28 38 27 61 21 23 30 17 32 16 . Prevalence of C. but given strain carries a few. Also makes beta-toxin (necrotic). perfringens C. perfringens produces cpe gene product as paracrystalline inclusion body released during sporulation.35 kDa protein (tissue damage and permeability change): 1) receptor mediated binding w/claudin + 50 kDa protein.

typhimurium): 105 . mortality is 15% if untreated. headaches. vomiting.48 hrs.Salmonellosis • Gram-negative enteric bacterium. chills. meats) and by food handlers • Colonization of intestinal epithelium • Two diseases: – Enterocolitis (most commonly by S. treatment . antibiotics • Prevention: Cooked food (70 ◦C for 10 min). transmission is from sources (eggs.000 people developed gastroentiritis after eating Schwan’s ice cream • Salmonella was isolated from 8 of 266 ice cream products (3%) • Trailers were transporting non-pateurized eggs immediately before carrying ice cream premix 17 . diarrhea and fever (2-3 days).none – Typhoid fever (Salmonella enterica serovar Typhi ): Septicemia leading to high fever that can last for several weeks. all strains are pathogenic.108 viable cells. disease onset within 8 . continuous shading of organism for months/years. monitor for carrier state among food handlers Salmonella Enteriditis: ice cream outbreak • Nation-wide outbreak • 224.

94-0.0 Temp. water and produce in developing countries. coli.0-8.colonization of the small intestine and verotxin production -> hemorrhagic diarrhea and kidney infection. uncooked and undercooked ground meat.diarrhea that afflicts young children • Enteroinvasive . 18 . classification of pathogens is based on toxin and diseases • Enterohemorrhagic (O157:H7) . diarrhea and urinary tract infection.invasive colon infection. survival in phagosomes.heat labile toxin.97 Pathogenic E.Escherichia coli • • • • • • • • Gram-negative rod non-sporeformer Flagellated facultative anaerobe generally harmless. pH: 6. coli • Some strains of E.:37oC Aw: 0. bloody diarrhea. in developing countries • Treatment and prevention: diseases are self-contained but antibiotics help. occasional epidemics • Enterotoxigenic (Travelers diarrhea) . immunity • Enteropathogenic .

E. 7 deaths Linked to school lunches Radish sprouts? Beef products of US origin? Sick people discriminated against School official blamed for outbreak committed suicide Campylobacter jejuni • Characteristics – High morbidity.000 cases. heat. acidic conditions. >9. low mortality – Sensitive to freezing – Survives in refrigerated foods – will not grow at <30◦C – Microaerophilic – Fragile organism.So why is it such a problem? 19 . sensitive to drying. and disinfectants. coli O157:H7 • • • • • • Japan.

20 . • Can Have "Y". non-sporeforming rod • 1-2 X 0. • Serotype Based on O & H Antigens. • Motile at 20-25 °C. "V". • Infective dose may be as low as 8 CFU/G.5 μM Long. small. • Produces haemolysin. or even streptococcal forms. but Not at 37 °C.Campylobacter jejuni • Disease – Onset 48-82 h following ingestion – Profuse diarrhea – Invasive – Survives phagocytosis Listeria monocytogenes • Gram+.

Linked with Mexican soft cheese .) • Listeric meningitis. 50% fatality rate.0. this drops to 30%. except in pregnant woman (who abort). monocytogenes without illness.15% to 2.disease syndromes • Low grade "flu-like" infection . monocytogenes.headache.Same phage type isolated from cases and processing plant . • Encephalitis • Psychosis • Infectious mononucleosis • Septicemia Los Angeles Mexican Soft Cheese .Cheese was alkaline phosphate positive 21 . J.93 Perinatal cases: 19 Still births 10 Postnatal deaths .0% of all perinatal mortality. Infection 13:187-193. • Up to 16% women carry L.49 Adult cases: 18 Deaths . drowsiness. • Perinatal infection .Plant's milk throughput exceeded capacity of pasteurizer .L. coma. If very young and old are excluded. 1986.not serious. (Larmont and Postlethwaite.

40% E. coli O157 .8% Campylobacter. 2005 / 54(14).42% April 15.352-356 22 . significant decreases in infections with: Salmonella .Incidence of reported cases and outbreaks of listeriosis in the United States. 1986-2004 Since 1996-98.31% Listeria .

Picornaviruses Polioviruses Coxsackieviruses A and B ECHO virus Enterovirus Hepatitis A 2. Papovaviruses Human BK and JC viruses 5. Adenoviruses Human adenoviruses types 1 . Reoviruses Reovirus Rotaviruses 3.33 23 .They are not alive but they are very bad! Human Intestinal Viruses with High Potential as Food Contaminants 1. Parvoviruses Human gastrointestinal viruses 4.

html Definition of Prions Prions are small. Circoviridae Hepadnaviridae – Hepatitis B Cystoviridae Reoviridae Coronaviridae. Poxviridae Adenoviridae. no detectable nucleic acids of any kind and no virus-like particles have been assoc iated with prions. To date. Flaviviridae (Hepatitis Caliciviridae (Hepatitis E. 24 . Myoviridae.mad-cow.cnrs-mrs.Genomyc and Physical Characteristics Type Nucleic Acid Description Envelope Virus Family Name Herpesviridae (EBV). Rhabdoviridae (Rabies) enveloped ds nonenveloped DNA ss ds/ss ds positive nonsegmented segmented nonsegmented positive ss negative reverse transcriptase nonenveloped enveloped enveloped nonenveloped enveloped nonenveloped enveloped enveloped RNA nonsegmented http://www. Prions cause scrapie and other spongiform encephalopathies of animals and http://igs-server. proteinaceous infectious particles that resist inactivation by procedures which affect nucleic acids. Papovaviridae Parvoviridae. Tectiviridae. Mumps). Norwalk) Retroviridae (HIV) Paramyxoviridae (Measles. Yellow fever) Picornaviridae (Hepatitis A).

htm 25 . if not entirely. generates PrPSc from the normal cellular isoform of the of a protein designated as the scrapie isoform of the prion protein. as yet undefined. Antibodies protect against prions in vivo http://www. A post-translational process. designated PrPC.Composition of Prions Prions are composed largely.hfsp. PrPSc.

Antibodies protect against prions in vivo FDC = follicular dendritic cells http://www. broilers. green onions 26 . Mexican queso fresco Norovirus and raw oysters Cyclospora and sprouts Salmonella Enteritidis and eggs. basil Hepatitis A and strawberries. almonds Salmonella Poona and cantaloupe MDR Salmonella Newport and ground beef. hot dogs.htm New pathogen x food combinations identified in outbreaks • • • • • • • • • E.hfsp. apple cider. coli O157 and ground beef. raw milk cheese Several Salmonellaserotypes and tomatoes Listeria monocytogenes and sliced deli turkey.

Food Technol.The new foodborne outbreak scenario Old focal scenario: • Acute large local outbreak • High dose. organic • CHANGES IN AWARENESS: computer databases • CHANGES IN DEMOGRAPHICS: larger sensitive populations • CHANGES IN PRIMARY FOOD PRODUCTION: scale of operation. 43(12)52-59. vacuum packaging • CHANGES IN HANDLING AND PREPARATION:home refrigeration. often egregious • Local solution New diffuse scenario: • Diffuse widespread outbreak • Low dose. global production • CHANGES IN FOOD PROCESSING TECHNOLOGY: chill. 1989) • CHANGES IN EATING HABITS: fresh. low attack rate • Increase in "sporadic" cases • Detected by lab-based subtype surveillance • Complex multistate investigation • "Industrial contamination event“earlier in food chain • Industry-wide implications Why do "new" pathogens emerge? (Lynton Cox. microwave • CHANGES IN THE MICROORGANISMS:plasmid 27 . high attack rate • Detected by group themselves • Local investigation • A local food handling error.

S.) • Frog Legs in France • Fruit and Vegetables in South Africa • Strawberries in U.Physical.S (Consumers Prefer 20:1) • Nation's Pride (U. Chemical.S.) Raw Chicken 28 . Irradiated Spices Do Not Require Specialized Labeling in the U. and Biological Preservation Methods IRRADIATED FOOD PRODUCTS • Spices (EC Banned Ethylene Oxide in 1981.

but does not include common salt. or chemicals applied for their insecticidal or herbicidal properties. or oils extracted from spices. acidulants 2. nitrites 4. sorbates OTHER 1. sugars. substances added to food by direct exposure to wood smoke. vinegars. spices. sulfites 29 . propionates 4. tends to prevent or retard deterioration. parabens 3. benzoates 2. p. antioxidants 3.” ANTIMICROBIAL CHEMICAL PRESERVATIVES MAJOR 1. 101.FDA DEFINITION OF A CHEMICAL PRESERVATIVE FDA-CFR TITLE 21.22 (A) “Any chemical that when added to food.

30 . They kill sensitive cells by forming pores in the membrane causing the leakage of cellular materials. and the depletion of transmembrane potential (Δψ) and/or the pH gradient.0 RCOOH RCOO+ + H+ RCOOH pH 7.Mechanism of Action RCOOpH 4.0 H+ RCOOH RCOO+ pH 7.0 H+ ATP ADP + Pi ATPase H+ Bacteriocins Bacteriocins are defined as ribosomallyproduced proteinaceous substances of bacterial origin that exhibit antimicrobial activity.

Multiple hurdle technology in food preservation 31 .

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