Milk composition and microbiology

Milk microbiology

In addition to being a nutritious food for humans, milk provides a favorable environment for the growth of microorganisms. Yeasts, moulds and a broad spectrum of bacteria can grow in milk, particularly at temperatures above 16°C.

Microbes can enter milk via the cow, air, feedstuffs, milk handling equipment and the milker. Once microorganisms get into the milk their numbers increase rapidly. It is more effective to exclude microorganisms than to try to control microbial growth once they have entered the milk. Milking equipment should be washed thoroughly before and after use rinsing is not enough. Bacterial types commonly associated with milk are given in Table 3.

Table 3. Bacterial types commonly associated with milk.






Pathogenic and spoilage


Staphylococcus aureus



S. agalactiae


S. thermophilus

Acid fermentation

S. lactis

Acid fermentation

S. lactis-diacetyllatic

Flavour production

S. cremoris

Acid fermentation

Leuconostoc lactis

Acid fermentation

Bacillus cereus



L. lactis

Acid production

L. bulgaricus

Acid production

L. acidophilus

Acid production


Acid production

Mycobacterium tuberculosis


Microbial growth

Microbial growth can be controlled by cooling the milk. Most microorganisms reproduce slowly in colder environments. Cooling milk also slows chemical deterioration.

The temperature of freshly drawn milk is about 38°C. Bacteria multiply very rapidly in warm milk and milk sours rapidly if held at these temperatures. If the milk is not cooled and is stored in the shade at an average air temperature of 16°C, the temperature of the milk will only have fallen to 28°C after 3 hours. Cooling the milk with running water will reduce the temperature to 16°C after 1 hour. At this temperature bacterial growth will be reduced and enzyme activity retarded. Thus, milk will keep longer if cooled.

Natural souring of milk may be advantageous: for example, in smallholder butter-making, the acid developed assists in the extraction of fat during churning. The low pH retards growth of lipolytic and proteolytic bacteria and therefore protects the fat and protein in the milk. The acidity of the milk also inhibits the growth of pathogens. It does not, however, retard the growth of molds.

Naturally soured milk is used to make many products, e.g. yoghurt, sour cream, ripened buttermilk and cheese. These products provide ways of preserving milk and are also pleasant to consume. They are produced by the action of fermentative bacteria on lactose and are more readily digested than fresh milk.

The initial microflora of raw milk reflects directly microbial contamination during production. The microflora in milk when it leaves the farm is determined by the temperature to which it has been cooled and the temperature at which it has been stored.

The initial bacterial count of milk may range from less than 1000 cells/ml to 106/ml. High counts (more than 105/ml) are evidence of poor production hygiene. Rapid tests are available for estimating the bacterial quality of milk.

Milk pasteurisation

Pasteurisation is the process used to destroy bacteria in milk. In pasteurisation, the milk is heated to a temperature sufficient to kill pathogenic bacteria, but well below its boiling point. This also kills many non-pathogenic organisms and thereby extends the storage stability of the milk.

Numerous time/temperature combinations are recommended but the most usual is 72°C for 15 seconds followed by rapid cooling to below 10°C. This is normally referred to as High Temperature Short Time (HTST) treatment. It is carried out as a continuous process using a plate heat-exchanger to heat the milk and a holding section to ensure that the milk is completely pasteurised. Milk is normally pasteurised prior to sale as liquid milk. Pasteurisation is used to reduce the microbial counts in milk for cheesemaking, and cream is pasteurised prior to tempering for buttermaking in some factories.

Batch pasteurisation is used where milk quantities are too small to justify the use of a plate heat-exchanger. In batch pasteurisation, fixed quantities of milk are heated to 63°C and held at this temperature for 30 minutes. The milk is then cooled to 5°C and packed.

The lower temperature used for batch pasteurisation means that a longer time is required to complete the process 30 minutes at 63°C, compared with 15 seconds a 72°C.

Effects of pasteurisation on milk

Pasteurisation reduces the cream layer, since some of the fat globule membrane constituents are denatured. This inhibits clustering of the fat globules and consequently reduces the extent of creaming. However, pasteurisation does not reduce the fat content of milk.

Pasteurisation has little effect on the nutritive value of milk. The major nutrients are not altered. There is some loss of vitamin C and B group vitamins, but this is insignificant.

The process kills many fermentative organisms as well as pathogens. Microorganisms that survive pasteurisation are putrefactive. Although pasteurised milk has a storage stability of 2 to 3 days, subsequent deterioration is cause by putrefactive organisms. Thus, pasteurised milk will putrefy rather than develop acidity.

In rural milk processing, many processes depend on the development of acidity, and hence pasteurisation may not be appropriate.

Milk sterilisation

In pasteurisation, milk receives mild heat treatment to reduce the number of bacteria present. In sterilisation, milk is subjected to severe heat treatment that ensures almost complete destruction of the microbial population. The product is then said to be commercially sterile. Time/temperature treatments of above 100°C for 15 to 40 minutes are used. The product has a longer shelf life than pasteurised milk.

Another method of sterilisation is ultra-heat treatment, or UHT. In this system, milk is heated under pressure to about 140°C for 4 seconds. The product is virtually sterile. However, it retains more of the properties of fresh milk than conventionally sterilised milk.

Microbiology of butter

Butter is made as a means of extracting and preserving milk fat. It can be made directly from milk or by separation of milk and subsequent churning of the cream. In addition to bacteria present in the milk other sources of bacteria in butter are: a) equipment, b) wash water, c) air contamination, d) packing materials, and e) personnel.


In smallholder buttermaking, bacterial contamination can come from unclean surfaces, the butter maker and wash water. Packaging materials, cups and leaves are also sources of contaminants. Washing and smoking the churn reduces bacterial numbers. But traditional equipment is often porous and is therefore a reservoir for many organisms.

When butter is made on a larger processing scale, bacterial contamination can come from holding-tank surfaces, the churn and butter-handling equipment.

A wooden churn can be a source of serious bacterial, yeast and mould contamination since these organisms can penetrate the wood, where they can be destroyed only by extreme heat. If a wooden churn has loose bands, cream can enter the crevices between the staves, where it provides a growth medium for bacteria which contaminate subsequent batches of butter. However, if care is taken in cleaning a wooden churn this source of contamination can be controlled. Similar care is required with scotch hands and butterworking equipment.

Wash water

Wash water can be a source of contamination with both coliform bacteria and bacteria associated with defects in butter. Polluted water supplies can also be a source of pathogens.


Contamination from the air can introduce spoilage organisms: mould spores, bacteria and yeasts can fall on the butter if it is left exposed to the air. Moulds grow rapidly on butter exposed to air.


Care is required in the storage and preparation of packaging material. Careless handling of packaging material can be a source of mould contamination.


A high standard of personal hygiene is required from people engaged in buttermaking. Personnel pass organisms to butter via the hands, mouth, nasal passage and clothing. Suitable arrangements for disinfecting hands should be provided, and clean working garments should not have contact with other clothes.

Control of microorganisms in butter

Salting effectively controls bacterial growth in butter. The salt must be evenly dispersed and worked in well. Salt concentration of 2% adequately dispersed in butter of 16% moisture will result in a 12.5% salt solution throughout the water-in-oil emulsion.

Washing butter does little to reduce microbiological counts. It may be desirable not to wash butter, since washing reduces yield. The acid pH of serum in butter made from ripened cream or sour milk may control the growth of acid-sensitive organisms.

Microbiological analysis of butter usually includes some of the following tests: total bacterial count, yeasts and moulds, coliform estimation and estimation of lipolytic bacteria.

Yeast, mould and coliform estimations are useful for evaluating sanitary practices. The presence of defect producing types can be indicated by estimating the presence of lipolytic organisms.

All butter contains some micro-organisms. However, proper control at every stage of the process can minimise the harmful effects of these organisms.

Standardisation of milk and cream

An adjustment of the fat content of cream is required, or if the fat content of whole milk must be reduced to a given level, skim milk must be added. This process is known as standardisation.

Microbial tests for raw and pasteurised milk

Tests are available to know the milk microbiological quality. Bacteria, coliform and somatic cell counts are frequently used.

Bacteria Count

The total bacteria count is the number of bacteria in a sample that can grow and form countable colonies on Standard Methods Agar after being held at 32°C for 48 hours.

Coliform Count

The coliform count is the number of colonies in a sample that grow and form distinctive countable colonies on Violet Red Bile Agar after being held at 32°C for 24 hours. Coliforms are generally only present in food that has been fecally or environmentally contaminated.

Somatic Cell Count

Somatic cells are blood cells that fight infection and occur naturally in milk. The presence of mastitis (an infection of the mammary gland) in the cow will increase the somatic cell count. The somatic cell count can be determined by direct microscopic examination or by electronic instruments designed to count somatic cells.

Antibiotics in milk

Antibiotics are used to treat mastitis infections. Cows under antibiotic treatment for mastitis infections may have antibiotic residues in their milk, therefore, milk from treated cows is either discarded or collected into a separate tank. Milk containing antibiotic residues is not used for human consumption. The legal standard, as defined by the Food and Drug Administration (FDA), requires that milk contain no detectable antibiotics when analysed using approved test methods. Regulatory action is taken against the farm with the positive antibiotic test.

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