August 25, 2011 Microbiology of IMFs Low Aw inhibit growth of mcg o Aw= 0.6- no microbial growth o Above Aw of 0.

.6, tolerant mcgs can grow (molds) o Aw- 0.86 (S.aureus) Heat during processing kills some mcg Length of storage lessen survivor Drying + solutes/ chemical preservatives Vacuum-packed/gas-impermeable Low Rh storage envt o Rh of envt %ERH of food (Aw x 100) o Mcgs produce water upon pathogenesis Freeze drying Lyophilization Dehydrates frozen food Vacuum sublimation o Liquid water Quick Less movement of solutes More expensive Freeze dried foods Very low Aw= no mcg growth No. of survivors o Enhanced by CHO, proteins etc in food o Decreases over time of storage Storage o Room ambient temp o Gas-impermeable containers o Prevents spontaneous lipid oxidation Rehydration o LESS microbial cells die

solid ice sublimation heat + vacuum

gas vapor

Freeze-drying Mcg Cultures Mild treatment Adding cryoprotectants (glycerol) Vacuum-packed FOOD PRESERVATION OF ADDITION OF ANTI-MCBAL AGENTS Anti-microbial Agents Chemicals added or naturally present Retard microbial growth or kill cells Preservatives Food-Additives Added intentionally in small amts o Anti-microbial agents o Antioxidants (-hydroxy toluene, BHT) o Emulsifiers o Flavors o Color o Bleaching agents o Sequestrants/ clarifying agent o Humectants (glycerols) o Foam inhibitors Use of Food Additives Maintain or improve nutritional quality Enhance keeping quality Make food more attractive Aid in food processing Additives in Food Preservation Success of chemical treatment of diseases Only GRAS can be used as food o (generally recognized as safe) o List by US FDA Ineffective chemotherapeutic agent Host toxicity Reasons for not allowing other additives Disguise faulty processing and handling methods o Addition of strong flavor (i.e Wasabe flavor)

 Lessen nutritive value Not safe for human consumption Anti-microbial Agents Static Agents- inhibit growth Cidal agents- kill Can be relative High [] cidal Low: static Diluted: enhance microbial growth MODE OF ACTION Interference with genetic system o Attach to part of genetic system (ribosomes) o No synthesis of protein- no new cells produced Damage to CW and CM o w/ site of attachment (chemically-active site) o dissolve lipids of membrane o lysis can occur o exposed membrane affected by adverse envt. Inhibition of enzymes o High [] of salts can limit enzymatic activity (alteration of pHloss of enzyme activity o Oxidizing agents--- cleaves sulfhydril (disulfide bridges) Binding of essential nutrients Factors dictating the effectiveness of chem. Preservatives Type of chemical and its [] o Higher [], more cells affected o Acceptable level Type of organisms and its physiological state o Spores < vegetative cells o Yeasts< molds o Stationary < log More resistant microbes during stationary log No. of organisms o High microbial load, high conc. of chemical composition of food and its pH o in solid food < in liquid o high OM < low OM

MORE SUSCEPTIBLE high OM

SEPTEMBER 1, 2011

Factors dictating the effectiveness of chem. Preservatives Composition of food and its pH o High pH= low acidity of preservative o Many preservative are weak acids o Dissociation tendency Ex. Sorbic acid H3O+ sorbate Temperature o High Temp= high effective (away from opt T) Time o Higher time of contact= high effective Chemical preservatives Benzoic acid/ Benzoates o Max. level: 0.1% o Antifungal agent o Antibacterial o Used in: Tomato catsup Salad dressings Fruit juices o pH~4 o MOA: Blocks oxidation of glucose and pyruvate (inactivates enzymes) Inhibits nutrient uptake Prevents endospore outgrowth Parabens o Ester of p-hydroxybenzoic acid o Methyl-, propyl-, heptylo Max. level: 0.1% o Anti-fungal (~100 ppm) o Anti-bacterial (10-4000 ppm) o G+>Go Undissociated at pH 8.3 o Less sensitive to pH Foods with pH 3 to 8 (acidic, beer and bakery pdts)

o MOA: same as benzoic ancid Sorbic Acid/ Sorbates o Ca, Na and K salts o Max level= 0.2% o Anti-fungal molds o Antibacterial (aerobic) LAB are resistant Clostridia and S. aureus can be inhibited o Used in cheeses, cakes, cured meats (+nitrite) o Used in fermented foods o MOA: Membrane disruption Inhibits enzymes Impairs nutrient uptake Propionic Acids/ Propionates o Ca or Na salts o Fungistatic (mold) used in bakery pdts o MOA: Lipophilics Membrane disruption Inhibits enzymes Impairs nutrient uptake Nitrites and Nitrates o K or Na salts o Inhibit mcbal growth o Stabilize red color of meat o Flavor devt o No3NO2o Used in curing formula of meats o Both oxidizing and reducing agent Low pH: NO3 3 HONO NO NITRITE NITROUS ACID NITROUS OXIDE o Effective: C. botolinum Endospore formation Vegetative cell growth S. aureus o Ineffective: LAB Enteric bacteria o MOA:

 Interfere with iron-sulfur enzymes (ferredoxin) o pH (4.5 and 5.5) o high salt o low Eh o Perigo factor- antibotulinal; nitrite + heat o Max level= 120 ppm o 40 to 80 ppm + sorbate Amines + NO2 NITROSAMINES Sulfur dioxide and sulfites o Gas/liq. SO2, Na, K salts of SO3, HSO3, S2O5 o Max level: 300 ppm o Used in dried fruits, lemon juice, molasses, wines, fruit juices Antimcbal (against LAB, YM) o Antioxidant o Not allowed in meat and other thiamine source o Enhances smell of food and color o MOA: strong reducing agent o SO2 and sulfurous acid: reacts with enzymes in cell Lactic Acid, Acetic Acid and their salts o MOA: lower Ph Enzyme activity Microbial growth o 4.8% Na lactate in pre-cooked meats Effective aginst: Yersenia, Listeria and Salmonella, L. monocytogenes Na diacetate against YM in breads and cakes Biologically-Based Preservation Systems Biocontrol Living mcgs (mcbal interference) o Added> controlled mcgs Direct products 1) Controlled acidification Lactic antagonism (LAB) Inhibitio ndue to: i. Decrease in pH due to acid produced by introduced mcg E.g. Microgard i. Cottage cheese with skim milk culture of ii. Propionibacterium freudenreichii subsp. Shermanii

iii. Acetate, propionic acid and proteins against G- and molds Bacon i. Produced by Wisconsin process ii. With Pediococcus acidilacti and other preservatives 2) Bacteriocins Anti-microbial proteins from LANB Narrow-spectrum i. E.g. nisin o By Lactococcus lactis o Lantibiotic (meso-and-3-methyl-lanthionine) o Against G+ (spore formers) o As food preservative: milk and dairy pdts, canned food, mayonnaise, baby foods, meat (replace NO2) o MOA: form spores in CM, inhibits a.a transport o Other uses: anti-mastitis dip Treatment of ulcer Surface sanitizing agents 3.) Pediocin Against Clostridia, Listeria 4.) Bacteriophages Viruses that infect bacteria Host-specific Some studies in: o Beef to control Pseudomonas o Poultry to control C. jejuni and Salmonella o Cheese to control L. monocytogenes and E. coli Factors to consider: o Lysis of host o Replicating host o Food constituents effect on attachment and lysis o Lytic bacteriophage used 5.) Modified Atmosphere Altering gaseous environment of foods High CO2 levels Low O2 levels 6.) Hypobaric storage 1 atm= 760 mmHg Stored in air under:

o Low pressure o Low temp o High humidity o Controlled ventilation Low O2 o No fat oxidation o Inhibits aerobic mcgs o Fruits/veg: 10-80 mmHg o Meats: 10 mmHg 7.) Vacuum packaging Air is removed before sealing Gas-impermeable containers (e.g. high-barrier plastic pouches) Heat-shrinking Air is squeezed out before sealing Pressure: 1 bar to 0.3-0.4 bar During storage: o O2 decreases o CO2 increases 8.) Modified Atmosphere Packaging (MAP) Hyperbaric process Flushing varying mixtures of CO2, N2 and O2 High o2 system o O2: up to 70% o CO2: 20-30% o N2: 0-20% Low O2 system o O2: 10% o CO2: 20-30% o N2: as necessary 8.) Controlled- Atmosphere Packaging or Storage (CAP, CAS) Controlled air in chamber More gas-impermeable packaging material o Aluminum foil laminates o Metal o Glass 9.) Equilibrium-Modified Atmosphere (EMA) Gas-permeable container Sealing the pack without gas alteration

MODE OF ACTION: CO2 Retard lag and log phase of microbial growth o 10% o Competitive inhibition of ethylene (senescence factor in fruits) Enzymatic decarboxylation Affects permeability of cell membranes o CO2 + H20 H2CO3 H+ + HCO32H+ + CO32 Accumulates in lipid Lowers intracellular pH Inhibits enzyme functions Factors that affect effectivity of CO2: Type of organisms (most sensitive-most resistant) o Pseudomonas o Aeromonas o Bacillus o Molds o Enterobacter o Enterococcus o Lactobacillus o Clostridium Temperature o High < low o more soluble CO2 in low temp pH o Acidic > neutral/ basic Pressure o More antimicrobial with pressure FOOD PRESERVATION BY IRRADIATION Radiation Kills mcgs With sufficient energy to eject electrons from an atom Shorter wavelength= higher energy; higher damage Ionizing- 2000 angstroms to ionize electrons - Ultra, xray, gamma ray Non-ionizing- non heat producing UV Light Absorbed by proteins and n.a

o Tymine dimmers of n.a (mutation) Poor penetrating power o Bottled water o Surface treatment of baked products (fruitcake) o Tables, equipment surface sterilization Microwave Rapid changes in electric currents o 915 million times per second Food molecules align with alternating current Intermolecular friction= heat Non-ionizing ray A. IONIZING 1.) Gamma rays Ionizing radioation o Breaks phosphodiester backbone of DNA o Produce free radicals From excited nucleus 60C or 136Cs o Atomic fission waste Excellent penetrating power Application rate: 1-100 grays (Gy), per min o 1 Gy= absorption of 1 joule per kg Radiation chamber (concrete-lead walls) Element source: emission in all directions 2.) Electron beams Accelerated o Beta rays (from radioactive source) o Cathode rays (from cathode like linear accelerators o poor penetrating power application rate: 10^3-10^6 Gy per sec directed beams amt. of energy is controlled xrays generated turned on/ off 3.) X-rays bombarding heavy metal target with cathode rays application to food: same with gamma-rays

3 Classes of Food Irradiation A. Radappertization radiation sterilization o commercially sterile pdts for pre-packaged foods (enzymes are inactivated) destroy pathogenic mcgs 12D concept Typical dosage level: 30-40 Gy Low salt, non-acid foods: 45-50 kGy Meats at subfreezing temp (-30C) Min, radiation doses (MRD) in kGy o Cured meat have nitrites (lower MRD) Do not follow 12D concept 2.) Radicidation Reduce numbers of viable, non-sporeforming pathogens to undetectable levels using standard methods Typical doses: 2.5-10 kGy 3.) Radurization Substantial reduction in number of spoilage mcgs Typical doses: 0.75- 2.5 kGy Effects of Radiation in food: Color Flavor Odor Loss of nutrients Free radicals from radiolysis o 3 H2O H + OH+ H2O2 + H2 *Methods for Reducing Side Effects of Ionizing Radiations to Foods o Addition of free radical acceptors o Reducing temp during treatment o Treatment under anaerobic condns o Reduction of moisture in food prior to irradiation *Factors influencing microbial resistance to radiation o Type of organism G+>G Enterococcus faecium, Entorococci Lactobacilli o Spores> veg cell C. botolinum type A P. larvae (aerobic)

o Yeast> mold o Complex< simple Viruses Deinococcus radiodurans can survive 15000 Gy Number of organisms o High number> low no. Composition of food o Complex> simple o Proteins, nitrite Presence or absence of O2 o Low O2>high O2 Moisture content o Low Aw> High Aw Temperature o Cold>hot Age of organism o Lag>log Storage stability of Irradiated Foods Radappertization-treated: o Microbiologically stable o Active food enzymes Radicidation/Raduration o Proper storage (low temp, etc.) Other Methods of Food preservation a.) High Hydrostatic Pressures (HHP) or Pascalization Non-thermal 100 to 1000 MPa for few min; 10000 atm Use of pressure cylinder Against vegetative cell Spores are activated Organoleptic properties not affected Meats, seafoods, jams anf=d fruit juices b.) Pulse Electric Fields (PEF) High electric fields in short pulses Non-thermal 5-55 kV per cm for 2 microseconds Electroporation of cell membrane Fruit juices and liquid eggs c.) Thermoultrasonification

 Spores are exposed to: o Ultrasonic waves: release Ca and other components in spores Food Preservation of Hurdle Technology Control of Microbial Growth Discruption of homeostasis o pH, temp, water content, etc. metabolic exhaustion o autosterilization during storage overcoming stress response o shock proteins Hurdle concept combination of various food preservation methods o temperature (high or low) o Aw o pH o Eh o Chem. Preservatives o Competitive organisms (LAB) Multitarget preservation