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Milk microbiology
standards for milk and milk product
Shivnam Rana
Msc. Microbiology (PAU)
L -2011- BS -251 – M
Introduction
• Complete food – Carbohydrates(Lactase),
Protein(Casein), Fats, Minerals
• due to: Complex biochemical composition
High water activity
• Excellent culture medium for the growth and
multiplication of microorganisms
Factors that influence survival and growth of
microorganisms
Intrinsic
• Nutrient availability
• pH
• Water availability
Extrinsic
• Temperature
• Atmosphere
Origin of microorganism in milk
• Commensal micro flora- teat skin, epithelial lining of the teat
canal, duct that conveys the milk from the mammary gland to
the teat orifice.
Staphylococcus, Streptococcus,Bacillus, Micrococcus,
Corynebacterium, coliforms
• Environmental contamination- soil, water equipment, dairy
farm area are reservoir for many food borne pathogens
Salmonella species,
L. monocytogenes
• Diseased animal
Mastitis- Staphylococcus (S.agalactiae) and
Streptococcus species
Lactobacillus
• Obligate homofermentative lactobacillus –
hexoses ferment lactic acid
• Growth at 45°C but not at 15°C
• Lactobacillus delbrueckii, lactobacillus helveticus,
lactobacillus salivarius and L. Acidophilus
• Facultative homofermentative lactobacillus-
hexoses ferment lactic acid or
glucoselimitation acetic acid, ethanol,
formic acid
• Grow at 15°C and show variable growth at
45°C
• Lactobacilluscasei,lactobacilluscurvatus,
lactobacillussakeiandlactobacillusplantarum.
• Obligate heterofermentative lactobacillus-
Hexoses ferment lactic acid+ acetic acid+
ethanol+ CO2
pentoses ferment lactic acid + acetic acid
• Lactobacillusfermentum,lactobacillusbrevisand
lactobacilluskeferi.
Benefit
• Resist weak acids of ph 3.5 to 4.5 resulting to
a yield of 90% lactic acid.
• Starter culture.
• Preservative for the production of cheese and
yoghurt.
Lactobacillus acidophilus
• Absorb B vitamins, vitamin K, fatty acids and
calcium.
• Reduce lactose intolerance.
• Increases the production of anti-allergy
cytokines, chemical messengers released by
cells of the immune system.
Lactobacillus bulgaricus
• Helps reduce symptoms of lactose intolerance.
• Provides the stomach and intestine with the
enzyme bacterial lactase that helps digest the
lactose in milk.
Enterococci
• Ubiquitous gram-positive, catalase-negative
• Ability to survive extreme ph, temperatures,
and salinity.
• Psychrotrophic nature, heat resistance and
adaptability to different substrates and growth
conditions
• Enterococci also occur in natural milk (or
whey) starter cultures
• Made by incubating it at 42–44 °C for 12–15 h
• Used for manufacturing of cheese ( artisan)
• Strains E. Faecalis and E. Faecium producing
enterocins
• Activity against listeria monocytogenes,
staphylococcus aureus, clostridium spp.,
Including clostridium botulinum and
clostridium perfringens, and vibrio cholerae
Bifidobacterium bifidum
• Optimum growth ph 6- 7.0
• Optimum growth temperature 37- 41°C,
maximum 43- 45°C, minimum 25- 28°C
• Produce acetic acid, lactic acid- 3:2
• Helping in digestion
• Strengthens the immune system
• To eliminate harmful bacteria
Lactobacillus rhamnosus
• Lactose-intolerant people to reduce the
inflammatory response that occurs when they
consume milk.
• Helps the immune system by stimulating the
production of antibodies and combating
pathogenic bacteria.
Lactose and galactose utilization by different lactic acid bacteria
Lactobacillus curd
• Milk is heated to a temperature of 30-40 °C
• Complex and heterogenous flora - L. Lactis , L.
Lactis subsp cremoris , S. Thermophilus , L.
Debrueckii subsp bulgaricus , L. Plantarum and
lactose fermenting yeast.
• Anaerobic respiration
Special attribute of Curd
• Dysentery and other gastrointestinal
disorders.
• Improve appetite , vitality and increase
digestibility.
yogurt
• Mixed starter culture – S. Thermophilus and lactobacillus delbrueckii or
lactobacillus bulgaricus.
• Ratio 1:1
• Fermentation :- lactose content of milk to yield lactic acid, CO2, acetic
acid, diacetyl, acetaldehyde
• Ph reduces :- around 6.5 to 4.5 due to the production of organic acids
• Initially streptococcus thermophilus ferments the lactose
• Lactobacillus bulgaricus, which is more acid tolerant, continues to ferment
the remaining lactose.
Lactase
Lactose glucose + galactose
Hydrolyzes
Benefit
• Easier digestibility,
• The ingested organisms
enhance bioavailability
of nutrients
• Ensure gastrointestinal balance,
• Promoting colon health
• Accelerates the healing of gastrointestinal
tract disorder
• Reduction in cholesterol level.
Cheese
Different cheese using different
strains
• Swiss cheese formation involves a late
propionic acid fermentation with ripening
done by Propioni bacteria shermanii.
• Blue cheeses are produced by Penicillium spp.
• Roquefort cheese is produced by using P.
roqueforti
• Camembert and Brie by using P. camemberti
and P. candidum.
Pathway for Citrate – positive strain of lactococcus and leuconostoc species
Changes in milk by
microorganism
Gas production:
• Fermentation occurs at faster rate, then raw
milk present a foamy layer on the upper
surface
• Air bubbles becomes entrapped and gas
becomes saturated throughout the body of
the milk
• Colliforms, clostridium and bacillus species.
Proteolysis
• Acid Proteolysis:
Milk whey separated, Milk taste will be soured, Micrococcus sp.
• Alkaline Proteolysis:
Milk whey separated and pH >6.9 (towards neutral and alkaline). Milk taste will be bitter
• Sweet curdling:
Bacillus cerus – it release enzymes “protease” which targets the casein. Bacteria use lactose
and convert into acids and aldehyde components. Milk is sweet in taste
• Slow Proteolysis:
Release of endogenous proteases in the milk, cause slow proteolysis of the milk.
• Proteolysis due to Anaerobic Bacteria:
Bacillus and clostridial species are heat resistant.
result into specific kind of smell names as butyrine smell
• Roppiness or sliminess:
Milk viscosity is increased, rope like structure is formed
Alcaligenes viscolactis
• Change in the color of milk:
Blue color ------------------------ pseudomonas synciani
Red color ------------------------- brubibacterium
erythrogenes,
sarcinia marcense
Yellow color --------------------- pseudomonas synxantha
Brown color --------------------- pseudomonas putrificians
Green color ---------------------- pseudomonas aurogenosa
Changes in Milk Fat
• Oxidation of unsaturated fatty acids:
Milk fat oxidation aldehydes + ketones + acids
• Oxidation imparts tallowy odors
• Hydrolysis of overall milk fat:
lipase
Fatty acids glycerols + free fatty acids
Hydrolysis
• Gives rise to putrefied odor (rotten egg like smell)
• Combined hydrolysis and oxidation
eg:- Proteus, Alcaligenes, Micrococcus
Change in the Flavor of Milk
Sour Flavour:
It is due to acidic changes in the milks:
Clean: Low contents of acids, Streptococcus lactis
Aromatic: streptococci and aroma- forming Leuconostoc sp. ,
moderated type of acidic components.
Sharp: coliform bacteria, clostridium species, volatile fatty acids,
high acidic contents
• Bitter Flavour:
It is due to alkaline changes in the milk.
• Potato-like Flavour:
Pseudomonas mucidolense
• Fishiness:
Acromian hydrophila, It is due to formation of tri-methyl amine
Polymerase chain reaction denaturing gradient gel electrophoresis (PCR-DGGE)
Fluorescent in situ hybridization(FISH)
ELISA
Prevention method
• Pasteurization
• Ultra-High Temperature- 138 °C to 150 °C for 4 to 15 seconds
• Steam under pressure- 115 to 118°C for 14 to 18 min, caned milk
• Radiation- gamma, UV rays
• Preservatives- sorbic acid, propionic acid, sugar, salt,
hydrogen peroxide
• Modified atmosphere packaging
• Refrigeration - 10 °C or low temperature
• Freezing- -17 to -18 °C
HACCP
Principle 1: Conduct a hazard analysis.
Principle 2: Determine the Critical Control Points
(CCPs).
Principle 3: Establish critical limit(s).
Principle 4: Establish a system to monitor control
of the CCP.
Principle 5: Establish the corrective action to be
taken when monitoring indicates that a
particular CCP is not under control.
Principle 6: Establish procedures for verification
to confirm that the HACCP system is working
effectively.
Principle 7: Establish documentation concerning
all procedures and records appropriate to
these principles and their application.
New bacteria discovered in raw milk
• Chryseobacterium oranimense, which can
grow at cold temperatures(7°C) and secretes
enzymes that have the potential to spoil milk.“
• C. haifense and C. bovis
laws
• Milk and milk product order, 1992
• Milk and milk product amendment regulation,
2009
Precautions
• The animal should be healthy and free from diseases.
• A healthy person should milk the animals. He should avoid sneezing, coughing,
etc., and must wear clean clothes.
• Milking vessels should be cleaned properly with chemicals or detergents that are
not injurious to health.
• Arrangements must be made in advance to immediately cool the milk to 4 ºC
within an hour of milking.
• The ingredients and cleaning agents used must be of the desired quality.
• There should be a provision for checking the quality, sampling and testing of milk.
Reference
• Beresford T P, Fitzsimons N A, Brennan N L and Cogan T M (2001) Recent advances
in cheese microbiology. International Dairy Journal 11 259–74.
• Ledenbach L H and Marshall R T 2009 Microbiological Spoilage of Dairy Products.
Compendium of the Microbiological Spoilage of Foods and Beverages 1-4419
• Nwamaka N T and Chike A E (2010) Bacteria population of some commercially
prepared Yoghurt sold in Enugu State, Eastern Nigeria. African Journal of
Microbiology Research 4 (10) 984-88.
• Papademas P and Bintsis T (2010) Food safety management systems (FSMS) in the
dairy industry: A review International Journal of Dairy Technology 63 14-07.
• Varga L (2007) Microbiological quality of commercial dairy products.
Communicating Current Research and Educational Topics and Trends in Applied
Microbiology
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Milk microbiology standards of milk and milk product22222 (2)

  • 1. Milk microbiology standards for milk and milk product Shivnam Rana Msc. Microbiology (PAU) L -2011- BS -251 – M
  • 2. Introduction • Complete food – Carbohydrates(Lactase), Protein(Casein), Fats, Minerals • due to: Complex biochemical composition High water activity • Excellent culture medium for the growth and multiplication of microorganisms
  • 3. Factors that influence survival and growth of microorganisms Intrinsic • Nutrient availability • pH • Water availability Extrinsic • Temperature • Atmosphere
  • 4. Origin of microorganism in milk • Commensal micro flora- teat skin, epithelial lining of the teat canal, duct that conveys the milk from the mammary gland to the teat orifice. Staphylococcus, Streptococcus,Bacillus, Micrococcus, Corynebacterium, coliforms • Environmental contamination- soil, water equipment, dairy farm area are reservoir for many food borne pathogens Salmonella species, L. monocytogenes
  • 5. • Diseased animal Mastitis- Staphylococcus (S.agalactiae) and Streptococcus species
  • 6. Lactobacillus • Obligate homofermentative lactobacillus – hexoses ferment lactic acid • Growth at 45°C but not at 15°C • Lactobacillus delbrueckii, lactobacillus helveticus, lactobacillus salivarius and L. Acidophilus
  • 7.
  • 8. • Facultative homofermentative lactobacillus- hexoses ferment lactic acid or glucoselimitation acetic acid, ethanol, formic acid • Grow at 15°C and show variable growth at 45°C • Lactobacilluscasei,lactobacilluscurvatus, lactobacillussakeiandlactobacillusplantarum.
  • 9. • Obligate heterofermentative lactobacillus- Hexoses ferment lactic acid+ acetic acid+ ethanol+ CO2 pentoses ferment lactic acid + acetic acid • Lactobacillusfermentum,lactobacillusbrevisand lactobacilluskeferi.
  • 10.
  • 11. Benefit • Resist weak acids of ph 3.5 to 4.5 resulting to a yield of 90% lactic acid. • Starter culture. • Preservative for the production of cheese and yoghurt.
  • 12. Lactobacillus acidophilus • Absorb B vitamins, vitamin K, fatty acids and calcium. • Reduce lactose intolerance. • Increases the production of anti-allergy cytokines, chemical messengers released by cells of the immune system.
  • 13. Lactobacillus bulgaricus • Helps reduce symptoms of lactose intolerance. • Provides the stomach and intestine with the enzyme bacterial lactase that helps digest the lactose in milk.
  • 14. Enterococci • Ubiquitous gram-positive, catalase-negative • Ability to survive extreme ph, temperatures, and salinity. • Psychrotrophic nature, heat resistance and adaptability to different substrates and growth conditions
  • 15. • Enterococci also occur in natural milk (or whey) starter cultures • Made by incubating it at 42–44 °C for 12–15 h • Used for manufacturing of cheese ( artisan)
  • 16. • Strains E. Faecalis and E. Faecium producing enterocins • Activity against listeria monocytogenes, staphylococcus aureus, clostridium spp., Including clostridium botulinum and clostridium perfringens, and vibrio cholerae
  • 17. Bifidobacterium bifidum • Optimum growth ph 6- 7.0 • Optimum growth temperature 37- 41°C, maximum 43- 45°C, minimum 25- 28°C • Produce acetic acid, lactic acid- 3:2 • Helping in digestion • Strengthens the immune system • To eliminate harmful bacteria
  • 18. Lactobacillus rhamnosus • Lactose-intolerant people to reduce the inflammatory response that occurs when they consume milk. • Helps the immune system by stimulating the production of antibodies and combating pathogenic bacteria.
  • 19.
  • 20.
  • 21. Lactose and galactose utilization by different lactic acid bacteria
  • 22.
  • 23. Lactobacillus curd • Milk is heated to a temperature of 30-40 °C • Complex and heterogenous flora - L. Lactis , L. Lactis subsp cremoris , S. Thermophilus , L. Debrueckii subsp bulgaricus , L. Plantarum and lactose fermenting yeast. • Anaerobic respiration
  • 24. Special attribute of Curd • Dysentery and other gastrointestinal disorders. • Improve appetite , vitality and increase digestibility.
  • 25. yogurt • Mixed starter culture – S. Thermophilus and lactobacillus delbrueckii or lactobacillus bulgaricus. • Ratio 1:1 • Fermentation :- lactose content of milk to yield lactic acid, CO2, acetic acid, diacetyl, acetaldehyde • Ph reduces :- around 6.5 to 4.5 due to the production of organic acids • Initially streptococcus thermophilus ferments the lactose • Lactobacillus bulgaricus, which is more acid tolerant, continues to ferment the remaining lactose. Lactase Lactose glucose + galactose Hydrolyzes
  • 26. Benefit • Easier digestibility, • The ingested organisms enhance bioavailability of nutrients • Ensure gastrointestinal balance, • Promoting colon health • Accelerates the healing of gastrointestinal tract disorder • Reduction in cholesterol level.
  • 28. Different cheese using different strains • Swiss cheese formation involves a late propionic acid fermentation with ripening done by Propioni bacteria shermanii. • Blue cheeses are produced by Penicillium spp. • Roquefort cheese is produced by using P. roqueforti • Camembert and Brie by using P. camemberti and P. candidum.
  • 29. Pathway for Citrate – positive strain of lactococcus and leuconostoc species
  • 30. Changes in milk by microorganism
  • 31. Gas production: • Fermentation occurs at faster rate, then raw milk present a foamy layer on the upper surface • Air bubbles becomes entrapped and gas becomes saturated throughout the body of the milk • Colliforms, clostridium and bacillus species.
  • 32. Proteolysis • Acid Proteolysis: Milk whey separated, Milk taste will be soured, Micrococcus sp. • Alkaline Proteolysis: Milk whey separated and pH >6.9 (towards neutral and alkaline). Milk taste will be bitter • Sweet curdling: Bacillus cerus – it release enzymes “protease” which targets the casein. Bacteria use lactose and convert into acids and aldehyde components. Milk is sweet in taste • Slow Proteolysis: Release of endogenous proteases in the milk, cause slow proteolysis of the milk. • Proteolysis due to Anaerobic Bacteria: Bacillus and clostridial species are heat resistant. result into specific kind of smell names as butyrine smell
  • 33.
  • 34. • Roppiness or sliminess: Milk viscosity is increased, rope like structure is formed Alcaligenes viscolactis • Change in the color of milk: Blue color ------------------------ pseudomonas synciani Red color ------------------------- brubibacterium erythrogenes, sarcinia marcense Yellow color --------------------- pseudomonas synxantha Brown color --------------------- pseudomonas putrificians Green color ---------------------- pseudomonas aurogenosa
  • 35. Changes in Milk Fat • Oxidation of unsaturated fatty acids: Milk fat oxidation aldehydes + ketones + acids • Oxidation imparts tallowy odors • Hydrolysis of overall milk fat: lipase Fatty acids glycerols + free fatty acids Hydrolysis • Gives rise to putrefied odor (rotten egg like smell) • Combined hydrolysis and oxidation eg:- Proteus, Alcaligenes, Micrococcus
  • 36.
  • 37. Change in the Flavor of Milk Sour Flavour: It is due to acidic changes in the milks: Clean: Low contents of acids, Streptococcus lactis Aromatic: streptococci and aroma- forming Leuconostoc sp. , moderated type of acidic components. Sharp: coliform bacteria, clostridium species, volatile fatty acids, high acidic contents • Bitter Flavour: It is due to alkaline changes in the milk. • Potato-like Flavour: Pseudomonas mucidolense • Fishiness: Acromian hydrophila, It is due to formation of tri-methyl amine
  • 38.
  • 39.
  • 40.
  • 41.
  • 42. Polymerase chain reaction denaturing gradient gel electrophoresis (PCR-DGGE)
  • 43. Fluorescent in situ hybridization(FISH)
  • 44. ELISA
  • 45.
  • 46. Prevention method • Pasteurization • Ultra-High Temperature- 138 °C to 150 °C for 4 to 15 seconds • Steam under pressure- 115 to 118°C for 14 to 18 min, caned milk • Radiation- gamma, UV rays • Preservatives- sorbic acid, propionic acid, sugar, salt, hydrogen peroxide • Modified atmosphere packaging • Refrigeration - 10 °C or low temperature • Freezing- -17 to -18 °C
  • 47.
  • 48.
  • 49. HACCP Principle 1: Conduct a hazard analysis. Principle 2: Determine the Critical Control Points (CCPs). Principle 3: Establish critical limit(s). Principle 4: Establish a system to monitor control of the CCP.
  • 50. Principle 5: Establish the corrective action to be taken when monitoring indicates that a particular CCP is not under control. Principle 6: Establish procedures for verification to confirm that the HACCP system is working effectively. Principle 7: Establish documentation concerning all procedures and records appropriate to these principles and their application.
  • 51.
  • 52. New bacteria discovered in raw milk • Chryseobacterium oranimense, which can grow at cold temperatures(7°C) and secretes enzymes that have the potential to spoil milk.“ • C. haifense and C. bovis
  • 53. laws • Milk and milk product order, 1992 • Milk and milk product amendment regulation, 2009
  • 54. Precautions • The animal should be healthy and free from diseases. • A healthy person should milk the animals. He should avoid sneezing, coughing, etc., and must wear clean clothes. • Milking vessels should be cleaned properly with chemicals or detergents that are not injurious to health. • Arrangements must be made in advance to immediately cool the milk to 4 ºC within an hour of milking. • The ingredients and cleaning agents used must be of the desired quality. • There should be a provision for checking the quality, sampling and testing of milk.
  • 55. Reference • Beresford T P, Fitzsimons N A, Brennan N L and Cogan T M (2001) Recent advances in cheese microbiology. International Dairy Journal 11 259–74. • Ledenbach L H and Marshall R T 2009 Microbiological Spoilage of Dairy Products. Compendium of the Microbiological Spoilage of Foods and Beverages 1-4419 • Nwamaka N T and Chike A E (2010) Bacteria population of some commercially prepared Yoghurt sold in Enugu State, Eastern Nigeria. African Journal of Microbiology Research 4 (10) 984-88. • Papademas P and Bintsis T (2010) Food safety management systems (FSMS) in the dairy industry: A review International Journal of Dairy Technology 63 14-07. • Varga L (2007) Microbiological quality of commercial dairy products. Communicating Current Research and Educational Topics and Trends in Applied Microbiology