CONTAMINATION, PRESERVATION AND

SPOILAGE OF MILK .

Name: Abu Al Farabi

Roll No: 04.
MS Fellow in Microbiology and Veterinary Public Health.
 MILK AND COMPOSITION OF MILK
Milk: Milk alludes to dairy animal’s milk, the lacteal discharge, for all intents and
purposes free from colostrums, got by the complete milking of one or more
sound cows and containing at least 3.25% fat and 8.25% solid not fat.

Composition of milk and dietary effect on milk composition:

1. 87% water, 4.2% lactose, 3% to 4% fat, 3.5% to 4.2% protein.
2. 30% of membrane material of milk fat.
3. It’s components are phospholipids (25%), cerebrosides (3%), cholesterols(2%
4. High fiber diets promote high butter fats by production of acetic acid.
5. High cereal or starch diets promote more protein by propionic acid .
6. Coagulated protein of milk is calcium casenate phosphate.
 NATURAL ANTIMICROBIAL PROPERTIES OF MILK
1.Lactoferrin: a. Iron binding glycoprotein, first isolated from cow’s milk.
b. Partly saturated with iron (5 to 30%), 0.2gm/L of milk.
c. Bacteriostatic effect against E.coli, Staphylococcus aureus.
2. Lactoperoxidase: a. Binds with thiocyanate to produce Hypothiocyanate.
3. Lysozyme: 1. Bovine milk contains both C lysozyme and G lysozyme.
2. Effective against E.coli in concert with immunoglobulin A.
3. Lysis of some species of Salmonellae with ascorbate.
4.N-acetyl beta-D glucosaminase: Indicator of tissue damage during mastitis.

Application : In china, farmers use sodium thiocyanate and sodium percarbonate

to activate lactoperoxidase activity of raw milk.
 PATHOGENS AND NON PATHOGENS OF MILK
Air borne contaminants: Pseudomonas, Flavobacterium, Alcaligenes, Some
coliforms, micrococcus , fungi.
Psychrotrophs: more detrimental to milk, able to grow at 5 degree celcius and
make up a large number of microbes in milk.eg: Pseudomonas fluorescens.
1. Spoilage of Refrigerated milk: Rancid, bitter and fruity flavor.
2. Spoilage of milk at 25 degree celcius: Souring flavor.

Sources of pathogens in milk:

*Animal itself: Mycobacterium bovis, Brucella species, Coxiella burneti.
*Human : Salmonella, Shigella, Environmental origin: Bacillus anthracis.
PRESERVATION TECHNIQUES OF MILK
 PASTEURIZATION OF MILK
1. LTLT : Low temperature. Long time 63 degree celcius 30 minutes.
2. HTST: High temperature. Short time 72 degree celcius 15 seconds.
3. UHT : Ultra high temperature., 130-135 degree celcius 1 seconds.

Pasteurization: (Inventor: Louis Pasteur)

4. Kills and removes all pathogens (100%).
5. Kills most non pathogens.(99%).
6. Surviving organisms are spore formers,
thermoduric and thermophilic organisms.
4. In case of batch pasteurization, alteration of milk taste is occured due to
extended holding time, target pathogen of UHT method: Coxiella burnetii
5. Target pathogen of HTST method : pathogenic bacteria ( spore of Clostridium sp.)
 STEPS OF PASTEURIZATION OF MILK
1. Milk chilling: 1. Reducing the temperatures (2 degree celcius- 5 degree celcius).

2. It causes arrest of bacterial growth and metabolism.

2. Pre-heating: * Chilled milk is heated to about 40 degree celcius.
* It is done to easy separation of butter fat for standardization.
3. Clarification: * Centrifugal clarifier is used to remove all soil and sediments .
* Removal of large solid particles through tubular metallic filters.
4. Standardization: Seperation of butter fat for making cream, butter, ghee.
5.Homogenization:Breaking down the milk fat droplets,prevent cream separation.
6. Heating section: Raising of temperature to kill spores of Clostridium botulinum.
7. Holding section: After heating, milk flows into the holding tubes.
 PASTEURIZATION OF MILK
1. Common thermodeuric organisms : Bacillus, Micrococci, Arthrobacter
Microbacteria, Enterococci (High temperature lactis), Corynebacteria.
2. No of pathogens in raw milk is 500,000 to 3,000,000 organisms / ml of milk.
3. While no of non pathogens in pasteurized milk is 2000 to 20,000 organisms
per ml of milk.
4. Differential methods to count non pathogens in milk such as :
 VIABLE COUNT METHOD ( POUR PLATE METHOD).
1. 1 ml of milk will be delivered to 9ml of sterile saline tube (10-1), mixed
and 1 ml diluted milk will be transferred from tube 1 to tube 2 (10-2),
2. 1 ml of milk will be delivered to an empty petridish, 1 ml of diluted milk
10-1, 10-2 to empty Petri dishes.
3. 9ml of molten nutrient agar will be poured to each Petri dish. Mix well .
4. Leave to solidify, then incubate at 37 degree celcius 48 hours.
5. Count no of colonies and multiply by dilution factor .
6. Determine no of viable non pathogens on milk. A plate containing more
than 300 colonies should not counted.
7. Cell morphology can be seen easily but it takes more time.
 DYE REDUCTION TEST
1. Methylene blue or resazurin dyes is uses in this test.
2. 1ml of dye will be mixed to 10 ml of milk, incubate it 1hr at 36 degree celcius.
3. If there is change in color, it means high bacterial load. It is not good quality
shorten expiration period. Reaction: Methylene blue – Leucomethylene blue
4. If no change in color (still blue), low bacterial load, good quality of milk,
expand expiration period.
 DIRECT MICROSCOPIC COUNT
1. Deliver 0.01 ml of milk on slide, air dry.
2. Stain with LW ( Levowitz- Weber) stain for 10 minutes.
3. Count bacterial clumps with the microscope.
Reading results:
4. Bacterial clump: Dark blue.
5. Protein, Leucocytes and somatic cells: light blue background.
6. Fat globules: Colorless.
7. Dirt: Brown.
8. Clump: Clump is one cell or group of cells of same type separated by a
distance from other clumps or cells.
 VACREATION AND SPOILAGE OF MILK
Vacreation: Pasturization of milk under reduced pressure by direct steam.
Spoilage of milk (Gas production): Chief gas formers are the coliform bacteria.
1. Species of Clostridium and Bacillus yield both hydrogen and carbon dioxide.
2. Yeasts, propionics and heterofermentative lactis that produce only C02.

Proteolysis of milk:
1. Acid proteolysis in which acid production and proteolysis occur together.
2. Sweet curdling by rennin like enzymes of bacteria at early stage of proteolysis
3. Slow proteolysis by intracellular enzymes of bacteria after their autolysis.
4. Residual proteolytic activity of heat stable proteinase.
 SPOILAGE OF MILK (Ropiness or Sliminess).
Stringiness: Caused by mastitis and in particular by fibrin from the cow’s blood.
Sliminess: Resulting from the thickness of cream. E.g. at the top of a bottle.
Non bacterial stringiness : This is due to thin flims of casein or lactalbumin .
Bacterial top ropiness : Milk is most ropy at the top by Enterobacter organism.
Bacterial surface ropiness :
1.Slimy material on surface of milk, develop best at refrigeration temperature.
2. Causal organism: Alcaligenes faecalis, Streptococcus lactis.
3. They reproduce within capsule and form long chains.
4. Ropiness is reduced if acidity is increased. (Lactic acid).
5. Acid is produced at mesophilic temperature.
 SPOILAGE OF MILK ( FLAVOR AND COLOR CHANGE)

1. Sour or acid flavor: Clean flavor by Streptococcus lactis, aromatic flavor by

lactic Streptococci with Leuconostoc and sharp flavor by volatile fatty acid of
coliform organism.
2. Bitter flavor: produced by proteolysis, lipolysis, fermentation of lactose.
Some cocci cause very bitter milk and Actinomycetes cause bitter musty flavor .
3. Burnt flavor: Caused by certain strains of Streptococcus lactis var maltigenes.

• Blue color of milk: Caused by Pseudomonas syncyanea with Streptococcus lactis

• Yellow color of milk: Caused by Pseudomonas synxantha, Flavobacterium spp.
• Brown color of milk: Caused by Pseudomonas putrefaciens and by the
enzymatic oxidation of tyrosine by Pseudomonas fluorescens.
 ADULTERANTS TEST OF MILK
1.Table sugar (Sucrose): Increased density of milk, not detected by lactometer
reading test. Ketose sugar+ Resorcinol= Red color ppt. (+ sugar content in milk)
2.Starch: Increases milk’s solid content. Uses of iodine> Milk solution turn into
blue black color> Due to the formation of Starch iodo complex.
3. Benzoic and Salicylic acid: Increases shelf life of milk. Uses of concentrated
Sulfuric acid and Fecl3> Buff colored and violet colored reaction products.
5. Soap: Increases the foaming of milk. Uses of Phenolphthaelin > Alkali will be
neutralized by the acidity of milk> Pink color will be observed.
6. Formalin: reacts with Sulfuric acid and Fecl3 > Gives a purple colored ring .
7. Ammonium Chloride: Uses of NaOH, NaOCl, and Phenol> Deep blue color.
THANKS TO EVERYONE.