Shelf-Life: Definition

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Raw Milk and Its Affect on Quality Don M. Breiner Manager, Quality & Regulatory Land O'Lakes Dairy Foods Carlisle, PA DMBreiner@landolakes.com Shelf-Life: Definition The period of time that a product can be kept under practical storage conditions and still retain acceptable quality. The practical & safe storage life of a food. A product can have 0 shelf-life if made from defective ingredients: Rancid, Oxidized, Bacterial, Strong Feed, Unclean - defects carry-over from raw milk -

Factors Influencing Milk Shelf Life Raw Milk Quality Raw milk bacteriological quality: - total counts, specific types & metabolic activity - heat resistant or thermoduric psychrotrophs Somatic cells counts & herd health: - related proteases, lipases & other enzymes Absorbed, chemical or cow related flavor defects - feedy, barny, lipid oxidized, rancid, medicinal Processing & Post-Processing Defects Post-Processing Contamination (Gram neg./gram pos.) Thermoduric psychrotrophs (Gram pos.) Process/Handling defects (e.g. cooked, light induces flavors) Raw Milk Quality - Influence on Shelf Life Flavor Defects - Non-Microbial Foreign or Chemical: Flavor/odor depends on contaminant - sanitizers, ointments, oils, exhaust fumes Chlorophenols medicinal/ bandage-like - phenols; sanitizers, pesticides, tannins (water) - chlorine/iodine; sanitizers, cleaning chemicals Cowy/Ketosis (chemical/biochemical): - unclean, medicinal (acetone) Other defects perceived as chemical - light induced; lipid oxidation

Flavor Defects - Non-Microbial Feed or Barn Related Flavors: Transmitted (absorbed): - lungs/rumen blood milk Flavor/odor notes of certain feeds - hay, silage; generally not objectionable Stronger/poor quality feeds - moldy feeds, apple pumice - onion/garlic, bitter weeds Barny/unclean from poor ventilation - manure/urine odors Raw Milk Quality - Influence on Shelf Life Flavor Defects - Non-Microbial Rancid (free fatty acids from fat lipolysis): Baby vomit, provolone cheese - over-agitation, starved pumps, air leaks, - homog. raw milk/mixing raw/homog. - influenced by equipment design - influenced by herd health Oxidized (auto-oxidation of unsat. fatty acids): Wet cardboard, old-oil, tallow, greasy mouth-feel - catalyzed by divalent cations - (copper, iron) - spontaneous related to feed or cow > high fat feeds (e.g., soy beans) > lack of anti-oxidants (e.g., vitamin E)

Bacteriological Quality Farm Sources Natural flora of the healthy udder (very low) Flora of mastitic cows (low to high) Exterior of the cow (low to medium) Dairy barn environment, air, water (low) Equipment milk contact surfaces (low to high) Increases due to bacterial growth: Milk residues on equipment Prolonged milking times Milk storage time/temperature Note: Bacterial types vary with source and conditions Total Bacteria Count Standards For Raw Milk (USA) Producer Raw Milk (PMO*): Commingled Raw Milk (PMO*): Industry Processing Standards: Industry Premium Standards: < 100,000/ml < 300,000/ml < 50,000/ml < 10,000/ml Milk can be stored for an additional 72 hours at the plant = potential for further growth (dependent on types, #s, temperature) PMO = Grade A Pasteurized Milk Ordinance

Bacteriological Quality Raw Milk Time Line Farm: Milk Picked-Up Every 48 Hours 100,000 cfu/ml limit Transport: To Dairy Plant in Hrs 300,000 cfu/ml limit Factors: Initial Load Types Temperature Growth Rates How are bacterial numbers influenced during time line? Plant Storage: Tanks Washed Every 72 Hrs 300,000 cfu/ml limit Raw Milk Quality - Influence on Shelf Life Bacteriological Quality Total bacterial counts at the time of processing: Defective raw milk will give defective processed milk Limited info when raw milk is acceptable - reports of >10 6 in raw before defects are apparent in pasteurized milk up to 14 d Types & metabolic activity: Bacteria differ in spoilage capabilities/enzymes Heat resistant enzymes from select organisms (?) - most published work on Pseudomonas spp. - most influence on UHT shelf-stabile products - limited information on pasteurized milk Heat resistant psychrotrophic bacteria (e.g., spore formers)

Bacteriological Quality Total bacterial counts at the time of processing: Defective raw milk will give defective processed milk Acid/Sour/Coagulated: - growth of lactic acid bacteria to large numbers; Lactococcus lactis most common - often associated with poor cooling (esp >70 F) - dirty equip/poor hygiene may be promoting factor Malty: - malt, caramel, grape-nut like odor /flavor - Lactococcus lactis var maltigenes Raw Milk Quality - Influence on Shelf Life Bacteriological Quality ID Raw Milk SPC/ml ------------- Days@7 C Pasteurized SPC/ml ------------------ Flavor 1 16,000,000 7 400 Slight bitter 14 27,000 Very bitter 2 27,000,000 7 100 Very bitter 14 300 Very bitter 3 11,000,000 7 200 Good 14 100 Good 4 5,000,000 7 1,500 Good 14 30,000 Unclean Source: Patel & Blankenagel, 1972. J. Milk & Food Technol. 35:203-206

Bacteriological Quality Heat Resistant Enzymes: Little published work on pasteurized and ultra-pasteurized milk Defects in pasteurized milk caused by these enzymes have not been described. It is likely that the period of storage is too short to produce noticeable off-flavors. Leloup et al found that bacterial proteases may influence milk flavor during prolonged storage of pasteurized milk, provided recontamination level is low. (Leloup et al 1980. Rapport EEG-onerzoekingskonstrakt 2935/79-04.2. EEC.) News Flash: Shelf-Life expectations are getting longer Opinion: Significant defects in pasteurized & UP milks unlikely when raw milk counts at processing are < 300,000 cfu/ml in most cases Raw Milk Quality - Influence on Shelf Life Heat Resistant Enzymes Testing for Heat Resistant Enzymes: No Practical Commercial Tests Tests researched: - Enzyme/substrate methods - ELISA based methods Testing for the Potential for Heat Resistant Enzymes: Total bacteria counts at the time of processing DMC, SPC, BactoScan Selective Gram-Negative counts Proteolytic counts

Microorganisms that Survive Pasteurization High Survival Rate: Microbacterium lacticum Micrococcus spp. Equipment Films, Cracks & Crevices Spore-formers (in spore phase) Bacillus Paenibacillus Clostridia Lower Survival Rate: Enterococcus spp. Lactobacillus spp. Streptococcus spp. Soil, Feeds, Manure = Soiled Teats, (Equipment) Sources of Thermoduric Bacteria (numbers > 20,000 in raw = illegal pasteurized milk) Dairy Farms: areas of persistent poor cleaning, old rubber parts, poor cooling, dirty cows & environment Bulk Milk Tankers: poorly cleaned tanks, bridges, milk hoses Plants: separators/clarifiers, poorly cleaned raw tanks Often used as an indicator of poor hygiene on the raw side (Thermoduric Counts: heat milk 63 C/30 min before SPC)

Testing Procedures Best indicator of raw milk s influence on shelf-life is it s quality at the time of processing. Standard Plate Count: Plate on SPC agar, incubate 32 C/48 hr Provides only historical data Direct Microscopic Counts: Can be used for bacteria and somatic cells Sensory Evaluation: Odor and appearance of milk Tasting raw milk not advisable - some lab pasteurize & taste Receiving Plant Storage C-L Tank Post-Mortem Raw Milk Quality - Influence on Shelf Life Testing Procedures Raw Milk Bacterial Tests: Laboratory Pasteurization Count: Lab pasteurize milk 63 C (145 F)/30 min Perform SPC (32 C/48 hr) on heated milk Alternative: Psychrotroph Count 7 C/10 d Preliminary Incubation Count (Raw): Incubate milk at 12.8 C (55 F) for 18 hr Perform a SPC compare to fresh raw SPC Coliform Bacteria Count: Plate milks on Violet red Bile Agar These tests are used as indicators of poor production methods on farm. No direct relationship to processed milk shelf-life has been found in the literature.

Relationship of Raw Milk Bacterial Tests to Shelf-Life Laboratory Pasteurization Count: - Detects bacteria that survive lab pasteurization - Does not determine whether these organisms are psychrotrophic unless Psychrotroph Count 7 C (45 F)/10 d) performed - Psychrotrophic Spore Count assay may be more useful** Preliminary Incubation Count (Raw): - Detects bacteria that grow well at 55 F, most are Gram-negative (?) - Gram-negative bacteria do not survive pasteurization - Does not indicate levels of heat resistant enzymes Coliform Bacteria Count: - Coliform Bacteria do not survive pasteurization Factors Influencing Shelf Life of Fluid Milk GN Post-Processing Contamination Most psychrotrophic bacteria associated with reduced shelf-life are Gram-negative rods (Pseudomonas spp.) Generally do not survive pasteurization, occur as postprocessing contaminants (i.e. dirty equipment,) Indicators of post-pasteurization contamination Very low levels of contamination can result in reduced shelf-life of fresh dairy products

Factors Influencing Shelf Life of Fluid Milk Sources of Thermoduric Psychrotrophs: Raw Milk - soil, grass, silage, concentrates, manure -cows, milking equipment, bulk tank - transport tankers, hoses - tanker CIP systems (?) - plant storage & handling Processing: - HTST/separator - filling machine - packaging material Sources of Bacteria in Raw Milk Pre-Milking Hygiene & Milk Let Down Recommendations Minimize water use Focus attention on teat surfaces, not udders Use an approved sanitizer (i.e. predip, spray) Assure complete coverage & contact time (> 30 sec) Provide 10-20 sec let-down stimulus e.g. teat message, fore stripping, teat drying Remove all dirt from teat surfaces e.g. wipe thoroughly dry & clean with clean towel From: Prepping Cows: Who Needs It? J. K. Reneau. NMC 2001 Proc Available at. http://www.nmconline.org/articles/prepping.pdf

Sources of Bacteria in Raw Milk Influence of Pre-Milking Hygiene on Milk Bacteria Counts 1 Anaerobic Teat Treatment Total Count 2 Coliform Count 2 Spores 2 No treatment 7,500 34 2 Water wash, wet 7,900 31 1.3 Water wash, dry 4,200 16 0.5 NaOCl wash, wet 4,100 38 0.7 NaOCl wash, dry 1,500 14 0.003 1 gram of teat soil of 100,000,000/gr into 30 lbs of milk = 7000/ml Counts as high as 100,000/ml of milk have been associated with dirty cows 3 1 From Bramley and McKinnon. 1990. The Microbiology of Raw Milk in Dairy Microbiology. 2 Counts per ml of milk. Mean of 30 trials for each treatment. Spores are related to cheese defect. 3 From J. Sumner. IDF Symposium Proceedings. 1996 Milk Under the Microscope Lactococcus species associated with poor refrigeration Staphylococcus aureus Streptococcus agalactiae

Somatic Cell Counts Legal Limits for Somatic Cells: USA 750,000 cells/ml Canada 500,000 cells/ml EU 400,000 cells/ml Herd Health Concerns: > 2-300,000 cells/ml Quality Concerns: proteases (e.g. plasmin), lipases = off-flavors in cheese and milk = reduced yields and texture defects in cheese Summary How Raw Milk Bacteria Influence Product Quality SPC of Producer Samples - most often does not milk at processing - depends on the types present, growth during storage SPC at the time of Processing - best indicator of product quality potential - counts >1 million suggest damaged, defective raw - counts > 300,000 may heat stabile enzymes Most bacteria in raw milk do not survive pasteurization - extended refrigeration selects for Gram-negatives SPC of fresh pasteurized milk mostly thermodurics (LPC) - limited growth of thermodurics in refrigerated raw