Chapter 1: Milk

1.1. Introduction

- Raw milk’ means milk produced by the secretion of the mammary gland of farmed
animals that has not been heated to more than 40 °C or undergone any treatment that has
an equivalent effect.
- ‘Dairy products’ means processed products resulting from the processing of raw milk or
from the further processing of such processed products.
Milk may be defined various ways. Chemically speaking, milk is a complex fluid in which more
than 100 separate chemical compounds have been found. Its major components are water, fat,
lactose, casein, whey proteins, and minerals (or ash) in amounts varying with the milk of various
species of animals. However, for any given species, the range of values for the constituents of
milk is fairly constant.
From a physiological standpoint, milk is the secretion of the normally functioning mammary
gland of the females of all mammals, which is produced for some time following parturition for
the nourishment of the young of the species during the initial period of growth.
In terms of physical chemistry, milk is an opaque, whitish fluid of multidisperse phases. The true
solution contains lactose, vitamins, acids, enzymes, and some inorganic salts. The colloidal
phase contains casein, calcium phosphate, and globular proteins. Fat exists in the form of an oil-
in-water type of emulsion, with fat globules varying from 0.1 to 22 µm in diameter.
As a food ingredient or consumed by itself, milk provides an excellent nutritional profile in the
human diet. Nutrition experts consider milk an exceptionally complete food because it contains
significant levels of required nutrients such as protein, fat, carbohydrates, minerals, and several
vitamins. Low-fat and nonfat milks are increasingly popular in fat-reduced and fat-free food
formulations.
Worldwide, milk of the cow is by far of more commercial importance than milk of any other
mammal. In the United States, the term “milk” legally refers to cow’s milk. Milk from other
species is labeled to indicate the type: sheep’s milk, goat’s milk, etc. Milk is the whole, clean
lacteal secretion of one or more healthy cows, properly fed and kept, excluding that obtained
within 15 days before calving and three to five days after.

Milk and dairy-based ingredients are used as components of many food products. Their
contributions consist of unique flavor, desirable texture, excellent nutritive value, and a widely
accepted “natural” image. In many instances, the success of the product in the marketplace is
significantly enhanced by incorporation of traditional functional ingredients familiar to the
consumer. Thus, dairy ingredients provide a consumer-friendly label on packaged foods.
Dairy ingredients contribute a number of characteristics critical to a food product. These include
the emulsifying and stabilizing ability of caseinates, the gelling properties of whey protein
concentrates and isolates, the water-absorption capacity of high-heat nonfat dry milk, and the
browning of lactose during heat processing. Furthermore, the crystallization characteristics of
lactose and the hydrolytic activity of the enzyme lactase are important in confectionery and
frozen products. In addition, butter flavor carryover can be achieved with enzyme-modified
butterfat and various cheese flavors imparted by enzyme-modified cheeses. Therefore, a food
developer can select an appropriate dairy-based ingredient to create certain desirable attributes in
foods. An understanding of the functional properties of dairy ingredients allows food

1
technologists to utilize their potential contribution to product characteristics to meet consumer
expectations.
1.1.2 Milk production and biosynthesis

Milk is the source of nutrients and immunological protection for the young cow. At parturition
fluid from the mammary gland known as colostrum is secreted-is yellowish colored, salty liquid
has a very high serum protein content and provides antibodies to help the newborn until its own
immune system is established. Within 2 hours the composition of colostrums returns to that of
fresh milk allowing to be used in food supply. Colostrum, the milk secreted immediately after
giving birth, is not considered milk from a legal standpoint.

1.1.3 Effects of Milk Handling on Quality and Hygiene

Cleanliness

The environment of production has a great effect on the quality of milk produced. From the food
science perspective, the production of the highest quality milk should be the goal. However, this
is sometimes not the greatest concern of those involved in milk production. Hygienic quality
assessment tests include sensory tests, dye reduction tests for microbial activity, total bacterial
count (standard plate count), sediment, titratable acidity, somatic cell count, antibiotic residues,
and added water.
The two common dye reduction tests are methylene blue and resazurin. These are both synthetic
compounds which accept electrons and change colour as a result of this reduction. As part of
natural metabolism, active microorganisms transfer electrons, and thus rate at which dyes added
to milk are reduced is an indication of the level of microbial activity. Methylene blue turns from
blue to colorless, while resazurin turns from blue to violet to pink to colourless. The reduction
time is inversely correlated to bacterial numbers. However, different species react differently.
Mesophilics are favoured over psychrotrophs, but psychrotrophic organisms tend to be more
numerous and active in cooled milk.

Temperature

Milk production and distribution in the tropical regions of the world is more challenging due to
the requirements for low-temperature for milk stability. Consider the following chart illustraing
the numbers of bacteria per millilitre of milk after 24 hours:

5°C 2,600
10°C 11,600
12.7°C 18,800
15.5°C 180,000
20°C 450,000

Traditionally, this has been overcome in tropical countries by stabilizing milk through means
other than refrigeration, including immediate consumption of warm milk after milking, by
boiling milk, or by conversion into more stable products such as fermented milks.
2
Mastitis and Antibiotics
Mastitis is a bacterial and yeast infection of the udder. Milk from mastitic cows is termed
abnormal. Its SNF, especially lactose, content is decreased, while Na and Cl levels are increased,
often giving mastitic milk a salty flavour. The presence of mastitis is also accompanied by
increases in bacterial numbers, including the possibility of human pathogens, and by a dramatic
increase in somatic cells. These are comprised of leukocytes (white blood cells) and epithelial
cells from the udder lining. Increased somatic cell counts are therefore indicative of the presence
of mastitis. Once the infection reaches the level known as "clinical' mastitis, pus can be observed
in the teat canal just prior to milking, but at sub-clinical levels, the presence of mastitis is not
obvious.

Somatic Cell Count (000's/ml) Daily Milk Yield (kg): 1st Lactation
0-17 23.1 29.3
18-34 23.0 28.7
35-70 22.6 28.0
71-140 22.4 27.4
141-282 22.1 27.0
282-565 21.9 26.3
566-1130 21.4 25.4
1131-2262 20.7 24.6
2263-4525 20.0 23.6
>4526 19.0 22.5

Antibiotics are frequently used to control mastitis in dairy cattle. However, the presence of
antibiotic residues in milk is very problematic, for at least three reasons. In the production of
fermented milks, antibiotic residues can slow or destroy the growth of the fermentation bacteria.
From a human health point of view, some people are allergic to specific antibiotics, and their
presence in food consumed can have severe consequences. Also, frequent exposure to low level
antibiotics can cause microorganisms to become resistant to them, through mutation, so that they
are ineffective when needed to fight a human infection. For these reasons, it is extremely
important that milk from cows being treated with antibiotics is withheld from the milk supply.
The withdrawal time after final treatment for various antibiotics is shown below:
Amoxcillin 60 hrs.
Cloxacillin 48 hrs.
Erythromicin 36 hrs.
Novobiocin 72 hrs.
Penicillin 84 hrs.
Sulfadimethozine 60 hrs.
Sulfabromomethozine 96 hrs.
Sulfaethoxypyridozine 72 hrs.

Anti-Microbial Systems in Raw Milk

There exists in milk a number of natural anti-microbial defense mechanisms. These include:
3
 Lysozyme - an enzyme that hydrolyses glycosidic bonds in gram positive cell walls. However,
its effect as a bacteriostatic mechanism in milk is probably negligible.
lactoferrin - an iron binding protein that sequesters iron from microorganisms, thus taking
away one of their growth factors. Its effect as a bacteriostatic mechanism in milk is also probably
negligible.
lactoperoxidase - an enzyme naturally present in raw milk that catalyzes the conversion of
hydrogen peroxide to water. When hydrogen peroxide and thiocyanate are added to raw milk, the
thiocyanate is oxidized by the enzyme/ hydrogen peroxide complex producing bacteriostatic
compounds that inhibit Gram negative bacteria, E. coli, Salmonella spp, and streptococci. This
technique is being used in many parts of the world, especially where refrigeration for raw milk is
not readily available, as a means of increasing the shelf life of raw milk.

1.1.4 Milk Grading and Defects

The importance of milk grading lies in the fact that dairy products are only as good as the raw
materials from which they were made. It is important that dairy personnel have a knowledge of
sensory perception and evaluation techniques. The identification of off-flavours and desirable
flavours, as well as knowledge of their likely cause, should enable the production of high quality
milk, and subsequently, high quality dairy products.
An understanding of the principles of sensory evaluation are neccessary for grading. All five
primary senses are used in the sensory evaluation of dairy products: sight, taste, smell, touch and
sound. The greatest emphasis, however, is placed on taste and smell.
The Sense of Taste
Taste buds, or receptors, are chiefly on the upper surface of the tongue, but may also be present
in the cheek and soft palates of young people. These buds, about 900 in number, must make
contact with the flavouring agent before a taste sensation occurs. Saliva, of course, is essential in
aiding this contact. There are four different types of nerve endings on the tongue which detect
the four basic "mouth" flavours -sweet, salt, sour, and bitter. Samples must, therefore, be spread
around in the mouth in order to make positive flavour identification. In addition to these basic
tastes, the mouth also allows us to get such reactions as coolness, warmth, sweetness,
astringency, etc.
The Sense of Smell

We are much more perceptive to the sense of smell than we are to taste. For instance, it is
possible for an odouriferous material such as mercaptain to be detected in 20 billion parts of air.
The centres of olfaction are located chiefly in the uppermost part of the nasal cavity. To be
detectable by smell, a substance must dissolve at body temperature and be soluble in fat solvents.

Note: The sense of both taste and smell may become fatigued during steady use. A good judge
does not try to examine more than one sample per minute. Rinsing the mouth with water between
samples may help to restore sensitivity.

Milk Grading Techniques

Temperature should be between 60-70° F (15.5-21° C) so that any odour present may be
detected readily by sniffing the container. Also, we want a temperature rise when taking the
sample into the mouth; this serves to volatize any notable constituents.

4
Noting the odour by placing the nose directly over the container immediately after shaking and
taking a full "whiff" of air. Any off odour present may be noted.
Need to make sure we have a representative sample; mixing and agitation are important.
Agitation leaves a thin film of milk on the inner surface which tends to evaporate giving off
odour if present.
During sampling, take a generous sip, roll about the mouth, note flavour sensation, and
expectorate. Swallowing milk is a poor practice.
Can enhance the after-taste by drawing a breath of fresh air slowly through the mouth and then
exhale slowly through the nose. With this practice, even faint odours can be noted.
Milk has a flavour defect if it has an odour, a foretaste or an aftertaste, or does not leave the
mouth in a clean, sweet, pleasant condition after tasting.
Characterization of Flavour Defects - ADSA
Lipolytic or Hydrolytic rancidity
Rancidity arises from the hydrolysis of milkfat by an enzyme called the lipoprotein lipase
(LPL). The flavour is due to the short chain fatty acids produced, particularly butyric acid.
LPL can be indigenous or bacterial. It is active at the fat/water interface but is ineffective unless
the fat globule membrane is damaged or weakened. This may occur through agitation, and/or
foaming, and pumping. For this reason, homogenized milk is subject to rapid lipolysis unless
lipase is destroyed by heating first; the enzyme (protein) is denatured at 55-60° C. Therefore,
always homogenize milk immediately before or after pasteurization and avoid mixing new
and homogenized milk because it leads to rapid rancidity.
Some cows can produce spontaneous lipolysis from reacting to something indigenous to the
milk. Late lactation, mastitis, hay and grain ratio diets (more so than fresh forage or silage),
and low yielding cows are more susceptible.
Lipolysis can be detected by measuring the acid degree value which determines the presence
of free fatty acids. Lipolytic or hydrolytic rancidity is distinct from oxidative rancidity, but
frequently in other fat industries, rancid is used to mean oxidative rancidity; in dairy, rancidity
means lipolysis.
Characterized: soapy, blue-cheese like aroma, slightly bitter, foul, pronounced aftertaste, does
not clear up readily
Oxidation
Milk fat oxidation is catalysed by copper and certain other metals with oxygen and air. This
leads to an autooxidation reaction consisting of initiation, propagation, termination.

RH --- R + H initiation - free radical

R + O2 ---- RO2 propagation

RO2 + RH --- ROOH + R

R + R --- R2 termination
R + RO2 --- RO2R
It is usually initiated in the phospholipid of the fat globule membrane. Propagation then occurs
in triglycerides, primarily double bonds of unsaturated fatty acids. During propagation, peroxide
derivatives of fatty acids accumulate. These undergo further reactions to form carbonyls, of
which some, like aldehydes and ketones, have strong flavours. Dry feed, late lactation,
added copper or other metals, lack of vit E (tocopherol) or selenium (natural antioxidates)
5
in the diet all lead to spontaneous oxidation. It can be a real problem especially in winter.
Exposure to metals during processing can also contribute.
Characterized: metallic, wet cardboard, oily, tallowy, chalky; mouth usually perceives a
puckery or astringent feel

Sunlight
Often confused with oxidized, this defect is caused by UV-rays from sunlight or flourescent
lighting catalyzing oxidation in unprotected milk. Photo-oxidation activates riboflavin which is
responsible for catalyzing the conversion of methionine to methanal. It is, therefore, a protein
reaction rather than a lipid reaction. However, the end product flavour notes are similar but
tends to diminish after storage of several days.
Characterized: burnt-protein or burnt-feathers-like, "medicinal"-like flavor

Cooked
This defect is a function of the time-temperature of heating and especially the presence of any
"burn-on" action of heat on certain proteins, particulary whey proteins. Whey proteins are
a source of sulfide bonds which form sulfhydryl groups that contribute to the flavour. The
defect is most obvious immediately after heating but dissipates within 1 or 2 days.
Characterized: slightly cooked or nutty-like to scorched or caramelized

Transmitted flavours
Cows are particulary bad for transmitting flavours through milk and milk is equally as
susceptible to pick-up of off flavours in storage. Feed flavours and green grass can be problems
so it is necessary to remove cows from feed 2-4 hrs before milking. Weeds, garlic/onion, and
dandelions can tranfer flavours to the milk and even subsequent products such as butter. Barny
flavours can be picked up in the milk if there is poor ventilation and the barn is not properly
cleared and cows breathe the air. These flavours are volatile so can be driven off through vacuum
de-aeration.
Characterization: hay/silage, cowy/barny

Microbial
There are many flavour defects of dairy products that may be caused by bacteria, yeasts, or
moulds. In raw milk the high acid/sour flavour is caused by the growth of lactic acid
bacteria which ferment lactose. It is less common today due to change in raw milk microflora.
In both raw or processed milk, fruity flavours may arise due to psychrotrophs such as
Pseudomonas fragi. Bitter or putrid flavours are caused by psychrotrophic bacteria which
produce protease. It is the proteolytic action of protease that usually causes spoilage in milk.
Malty flavours are caused by S.lactis var. maltigenes and is characterized by a corn flakes
type flavour. Although more of a tactile defect, ropy milk is also caused by bacteria, specifically
those which produce exopolysaccharides.

Miscellaneous Defects

astringent
chalky
chemical/medicinal - disease - associated or adulteration
flat - adulteration (water)
6
 foreign
salty - disease associated
bitter - adulteration
Milk flavour is graded on a score of one to 10. Some flavour defects, even if only slightly
present, can decrease the score drastically. The following are suggested flavour scores for milk
with designated intensities of flavour defects.

Flavour Criticisms Slight Intensity of Defect Definite Pronounced

Astringent 8 7 5
Barny 7 5 3
Bitter 7 5 3
Cooked 9 8 6
Cowy 6 4 1
Feed 9 7 5
Flat 9 8 7
Foreign 5 3 0
Garlic/onion 5 3 1
High acid 3 1 0
Bacterial 5 3 0
Lacks Freshness 7 5 3
Malty 7 5 3
Oxidized 7 5 3
Rancid 7 5 3
Salty 8 6 4
Unclean 7 5 3

7
 1.2: PHYSICO-CHEMICAL PROPERTIES OF MILK

1.2.1. Constituents of Milk

Milk is composed of water, milkfat, and MSNF. The MSNF consists of protein, lactose, and
minerals. These solids are also referred to as skim solids, or serum solids. The term total solids
refers to the serum solids plus the milkfat.
The major components of commercial raw milk are illustrated in Figure 2-1. On a dry basis, the
composition of milk solids is shown in Figure 2-2.

Fig. 2-1. Gross composition of milk, showing major constituents.

Fig. 2-2. Composition of milk solids of whole milk.

Major constituents
The major constituents of milk vary more widely in individual cow’s milk than in pooled market
milk. Factors affecting the milk such as breed of cow, intervals of milking, stages of milking,
different quarters of udder, lactation period, season, feed, nutritional level, environmental
temperature, health status, age, weather, oestrum, gestation period, and exercise are known to

8
cause variations in fat, protein, lactose, and mineral levels in milk derived from individual cows.
In general, these variations tend to average out but display an interesting seasonal pattern in
commercial milk used by food processors, which may have an important impact on properties of
the finished products. An approximately 10% variation in fat and protein is observed in milk
received in July and August (lowest level) compared to that received in October and November
(highest level).
Variations in protein and fat during the year affect yogurt and cheese manufacture and whey
protein production. For example, appropriate adjustments, such as fortification with additional
nonfat milk solids, are necessary in yogurt production to ensure uniform viscosity throughout the
year.

Milk proteins. Caseins make up approximately 80% of milk proteins. The remaining proteins
are classified as whey proteins. Milk proteins and their concentration in milk are shown in Table
2-1
Table 2-1. Milk Proteins

Besides having a biological and nutritional role, caseins and caseinates are important because of
their structure, charge, and physical properties. Caseins become insoluble when the milk is
acidified and the pH is reduced to 4.6, while the whey proteins remain in solution. Caseins exist
in milk as particles called micelles, which are made up of calcium phosphate and casein
complexes. The micelles are spherical particles varying in size from 50 to 200 nm and containing
thousands of protein molecules. Caseins are further divided into αs1, αs2, β, and κ fractions. A γ-
fraction is derived from the breakdown of β-casein by inherent proteolytic enzymes of milk.
Each of the casein fractions exhibits distinct chemical properties. For example, αs1 and αs2
caseins have eight and 10–13 phosphoserine units, respectively. β-Casein has five phosphoserine
units. All the casein fractions except κ-casein are precipitated by calcium, whereas κ-casein,
9
which contains one phosphoserine unit, is not calcium-sensitive. Only κ-casein contains a
carbohydrate moiety. Calcium (30 mM) and phosphate (20 mM) are complexed with αs1, αs2, and
β- caseins, whereas κ-casein stabilizes the colloidal particles (micelles) by surface binding.
During cheesemaking, the stabilizing κ-casein is cleaved by the enzyme rennin, resulting in
destabilization of the micelle and subsequent curd formation.
Whey proteins are located in solution in the serum phase of milk along with minor proteins and
enzymes, including lactoperoxidase, lactotransferrin, lysozyme, glycoprotein, serum transferrin,
and degradation products derived from casein.
Caseins and whey proteins are distinguishable from each other by their physical and chemical
characteristics (Table 2-2). Caseins have a distinct, disordered molecular structure that lacks
stabilizing disulfide bridges. This characteristic structure makes the casein fraction precipitate in
acidic conditions as well as in the presence of di- and polyvalent ions of various salts. Casein
molecules exist naturally in an open and extended state. Therefore, heat has little or no effect on
them. However, severe heat treatment results in detachment of phosphate groups as well as
formation of brown pigments by interaction with milk sugar (the Maillard reaction).

Table 2-2. Major Physical and Chemical Differences in Milk Proteins

Whey proteins have a relatively more ordered molecular structure, which contains disulfide
linkages. Accordingly, they are not vulnerable to precipitation under acidic conditions or by
polyvalent ions. Like other globular proteins, they can be heat-denatured, resulting in gel
formation. β-Lactoglobulin complexes with κ-casein in milk subjected to rigorous heat treatment.
In food systems, milk proteins contribute to properties of the final product. Table 2-3
summarizes the major functional characteristics of milk proteins.

10
Table 2-3. Functional Characteristics of Milk Proteins

Milkfat. The fat in milk occurs in microscopic globules in an oil-inwater type of emulsion. The
globules vary in size from 0.1 to 22 μm in diameter. The lipid content of milkfat is 97–98%
triacylglycerols, 0.2–1% phospholipids, 0.2–0.4% sterols, and traces of fatty acids, as well as
vitamins A, D, E, and K. Milkfat is made up of 65% saturated, 32% monounsaturated, and 3%
polyunsaturated fatty acids. It contains 7% short-chain fatty acids (C4–C8), 15–20% medium-chain fatty acids
(C10–C14), and 73–78% long-chain fatty acids (C16 or higher). The cholesterol content of whole milk (3.3%
fat) and skim milk is 14 mg/100 ml and 2 mg/100 ml, respectively. The functional properties of milkfat are
attributed to its fatty acid make-up.
Milk sugar. Lactose is the major carbohydrate of milk, occurring at a 4.5–4.9% level. It consists of two forms
(α and β) that differ in their sweetness and solubility. The α-form is less soluble (70 g/L at 15°C)
than the β-form. Crystallization is important in the manufacture and utilization of several dairy
ingredients. An equilibrium mixture of α- and β-lactose, formed by mutarotation, exhibits a
solubility of 170 g/L of water. β-Lactose is slightly sweeter than the α-form. Overall, in
sweetening power, lactose is only 16–33% as sweet as sucrose. The sweetening power is
concentration dependent.

Minor and Trace constituents

The typical composition of major milk minerals is shown in Table 2-4. Mineral concentration in
milk is related to physical-chemical equilibria, which are important in processing, nutritive
value, and shelf life of dairy products. Minerals include chloride, PO42+, and citrates of K+, Na+,
Ca2+, and Mg2+. Their concentration is <1% in milk, but they are involved in heat stability and
alcohol coagulation of milk, age-thickening of sweetened condensed milk, feathering of coffee
cream, rennin coagulation, and clumping of fat globules upon homogenization. The calcium level
of milk influences the firmness of curd during cheesemaking.

11
Ash, the white residue after incineration of a given weight of milk, is used as a measure of the
mineral content of milk. It is not identical to milk mineral level because of decomposition and
volatilization of certain minerals due to heat. Ash contains carbonates derived from organic
constituents; sulfates from proteins; phosphate, partly from casein, which may contain
approximately 1.62% phosphate; and chloride, which is partly lost (45–50%). Citric acid is
completely lost. The average ash content is 0.70%, which is equivalent to 0.90% minerals.
All the minerals considered essential for human nutrition are found in milk. They exist in milk in
different states. Sodium, potassium, and chloride are ionic forms and in true solution. They
permeate freely across the membrane during ultrafiltration and electrodialysis of milk and whey.
Calcium, magnesium, inorganic phosphorus, and citrate exist in both colloidal and diffusible
forms, depending on the pH of milk. Approximately 20–30% of diffusible calcium and
magnesium exists as free ions and the remainder as salts of citrate and phosphate. As the pH of
milk drops, the colloidal form is converted progressively to the ionic form. At pH 4.4, most of
the minerals are in diffusible form.
Table 2-4. Major Mineral Composition of Milk

By lowering the pH of milk or whey, protein concentrates of low mineral content and different
mineral ratios can be produced. Trace elements are those constituents found in the parts per
million level in milk. Table 2-5 shows their levels.

12
Table 2–5. Trace Minerals of Milk

In addition, several nonprotein nitrogen compounds, vitamins, and some organic materials are
present in milk (Table 2-6).
•Urea is responsible for almost all of the seasonal variation in the heat stability of milk. The
concentration of milk urea is controlled by the level of urea in the blood, which is directly related
to diet.
•As mentioned earlier, lipoprotein lipase catalyzes the hydrolysis of triglycerides to free fatty
acid. This reaction causes dairy products to have soapy, bitter, rancid, and unclean flavors. A
great deal of lipoprotein lipase is present in freshly drawn milk, and under certain conditions,
will spoil the milk within a few minutes. The fat globule membrane acts as a physical barrier to
lipase, and it is also readily inactivated by heat. Spontaneous lipolysis is influenced by stage of
lactation, season, and diet.
•Lactoperoxidase is another enzyme in milk present at high concentrations. It catalyzes the
oxidation of unsaturated fatty acids leading to the development of oxidized flavors. However, if
thiocyanate and peroxide are supplemented in milk, lactoperoxidase acts as a powerful
bacteriocide that can kill coliforms, Salmonellae, Shingellae, and Pseudomonads. This process
has been widely used for short-term preservation of milk in developing countries where
refrigeration is scarce.
•Xanthine oxidase is also present in milk and can catalyze non-specific oxidation of dairy
products. Other that, its overall significance is low.
•Alkaline phosphatase is almost completely inactivated by pasteurization, and it is therefore used
as an index of the efficiency of such heat treatments.

13
Table 2–6. Other Trace Components of Milk

1.2.2. Physical properties of milk

Color
The color of milk is perceived by consumers to be indicative of purity and richness. Its white
color is due to the scattering of reflected light by the inherent ultramicroscopic particles, fat
globules, colloidal casein micelles, and calcium phosphate. The intensity of white color is
directly proportional to the size and number of particles in suspension. Homogenization
increases the surface area of fat globules significantly as a result of the breakup of larger
globules. Accordingly, homogenized milk and cream are whiter than their unhomogenized
counterparts. Lack of fat globules gives skim milk a blue tinge.
Cow’s milk contains the pigments carotene and xanthophyll, which tend to give a golden yellow
color to the milkfat.
Flavor
The flavor of milk is a property difficult to define, but there is no doubt that taste and aroma are
critical to the assessment of milk. Flavor constitutes a critical criterion of quality for the

14
consumer. It is a sensory property in which odor and taste interact. The sweet taste of lactose is
balanced against the salty taste of chloride, and both are somewhat moderated by proteins. This
balance is maintained over a fairly wide range of milk composition even when the chloride ion
level varies from 0.06 to 0.12%. Saltiness can be detected by sensory tests in samples containing
0.12% or more of chloride ions and becomes marked in samples containing 0.15%. Some
workers attribute the characteristic rich flavor of dairy products to the lactones, methylketones,
certain aldehydes, dimethyl sulfide, and certain short-chain fatty acids.
Although milk has a clean, pleasantly sweet flavor, it is quite bland, and therefore any off-flavors
are readily discernible. Off-flavors result when the balance of flavor compounds is altered by
microbiological action, dairy farm or processing conditions, or chemical or biochemical
reactions.
Density and Specific gravity
The density of milk with 3–5% fat averages 1.032 g/cm3. Accordingly, the weight of 1 L of milk
is 1.03 kg. To convert the weight of milk from kilograms per liter to pounds per gallon, the
number is multiplied by 8.34.
The average specific gravity of milk at 15.5°C (60°F) is 1.032. It varies from 1.028 to 1.035 in
commercial milk. The specific gravities of milkfat (0.93), MSNF (1.62), and water (1.0)
determine the specific gravity of milk. Specific gravity is increased by the removal of fat and
lowered by the addition of water.
Surface tension
Surface properties are involved in adsorption phenomena and the formation and stability of
emulsions. They are relevant to creaming, fat globule membrane function, foaming, and
emulsifier use in dairy products. Normal cow’s milk has an inherent surface activity. Its surface
tension approximates 70% of that of water. The surface tension of whole milk is usually about
50–52 mN/m (or dyn/cm) at 20°C. The surface tension of skim milk is 55–60 mN/m. For cream,
it is approximately 46–47 mN/m. Casein, along with the proteolysis products protease-peptones,
is largely responsible for the surface activity. Whey proteins make little contribution. Fat reduces
surface tension by a physical effect. Lactose and most of the salts tend to raise it when they are
present in true solution.
Surface tension decreases as milk temperature rises. Processing treatments such as heating,
homogenization, and shear tend to increase surface tension.
Foaming
The formation of stable foam depends upon two main factors. First, the lowering of the surface
tension allows the gathering and spreading of the surface-active components into thin films.
Second, the films must be sufficiently elastic and stable to prevent the coalescence of the gas
cells. A stable foam is thus formed when the surface tension of the liquid is not great enough to
withdraw the film from between the gas cells and when the stabilizing agent has great internal
viscosity.
Foaming of milk is at a minimum at 30–35°C. At 60°C, the foam volume is independent of the
fat content. Below 20°C and above 30°C, the foaming tendency appears to increase. Fat tends to
15
stabilize the foam formed below 20°C, for instance, during churning. Skim milk produces
slightly more stable foam above 30°C than whole milk or light cream.
Foaming properties affect handling of milk products and how dairy-based ingredients are
incorporated into other products.
Viscosity
Whole milk and skim milk display viscosities of 2.0–2.1 and 1.5–1.8 cP (or mPa/sec) at 20°C,
respectively. Whey has a viscosity of 1.2 cP. The viscosity of milk and cream creates the
impression of “richness” to the consumer. From an organoleptic standpoint, viscosity contributes
to mouthfeel and flavor release.
The casein micelles of milk contribute more to the viscosity of milk than any other constituent.
Viscosity varies not only with changes in the physical nature of fat but also with the hydration of
proteins. Alterations in the size of any dispersed constituents result in viscosity changes. The fat
contributes less than casein but more than whey proteins. When fat globules are greatly
subdivided by homogenization, an increase in viscosity is observed. The viscosity of skim milk
decreases on heating to 62°C, after which it increases, apparently due to changes in protein
hydration. An increase of temperature causes a marked reduction of viscosity. For example, at
20°C, milk is about half as viscous as at 0°C and at 40°C is approximately one-third of the value
at 0°C.
Specific heat
The specific heat of milk products is a function of their composition. The values for whole milk,
skim milk, 40% cream, butter, and whey at 15°C are 0.93, 0.95, 0.68, 0.53, and 0.97 BTU/lb•°F,
respectively (3.89, 3.97, 3.35, 2.21, and 4.06 kJ/kg•K, respectively).
Electrical conductivity
Current passes through the milk by virtue of the activity of its ionic mineral constituents, of
which the chloride ions carry 60–68% of the current. There is therefore a close correlation
between the electrical conductivity of milk and its chloride content. The electrical conductivity
of normal milk corresponds to that of approximately 0.25% sodium chloride solution (w/w) and
ranges from 45 to 55 A•v–1m-1 (8) or 45–55 x 10–4 mho. Conductivity of milk is the basis of the
new ohmic process for sterilizing milk.
Lactic acid accumulates as a result of fermentation during the manufacture of yogurt and
fermented dairy products, converting calcium and magnesium to ionic form and thereby
increasing the conductivity reading. Thus, the progress of fermentation can be followed by
increases in the conductivity of the yogurt base. Also, demineralization of whey and its fractions,
leading to loss of ionic minerals, is monitored using a conductivity meter.
Freezing point
The freezing point of milk is generally expressed as degrees Hortvet (H), reflecting the
commonly used Hortvet crysoscope procedure. Determination of freezing point is a parameter
widely used in the industry for detection of adulteration of milk with water. The freezing point of
milk has a relatively narrow range (from –0.520 to –0.560°C, the average being –0.540°C). It

16
corresponds to the freezing point of an 0.85% sodium chloride solution (w/w). Addition of water
raises the freezing point, and readings above –0.520 support strong suspicion of watering the
milk. As little as 3% water added to milk can be detected by this method.
Lactose and chloride are the major milk constituents responsible for 70–80% of the overall
depression in the freezing point of milk.
Boiling point
A solution boils at a higher temperature than does the pure solvent, according to the
concentration of the dissolved substance. The boiling point of milk is 100.17°C. The milk
constituents in true solution are mainly responsible for the elevation of the boiling point above
100°C. Elevation of the boiling point is based on the same principles as depression of freezing
point. However, for detecting added water, the freezing point method is far superior on the
grounds of accuracy and convenience.
Refractivity
The refraction of light by a solution is a function of the molecular concentration of the solute in
solution. Each solute maintains its own refractivity, and the refractive index of a mixture is that
of the total of the refractive indices of the substances plus that of the solvent.
The components of milk contributing to its refractive index in descending order of importance
are water, proteins, lactose, and minor constituents. Whey proteins are more important than
casein. The refractive index of milk at 20°C is 1.3440–1.3485.
The refractive index of butterfat is distinct enough to indicate its authenticity.
Redox Potential
The oxidation-reduction or redox potential (Eh) of a redox system at 25°C is given by

where E0 = standard redox potential (a characteristic of each system, dependent on temperature

and especially on pH); n = number of electrons per molecule involved in the oxidation-reduction
reaction; [Ox] and [Red] = molar concentrations of the compound concerned in the oxidized and
reduced forms, respectively. Equation only holds true for reversible reactions. For n = 1, an
increase of Eh with 0.1 V thus corresponds with an increase of the relative concentration of the
oxidized form from, e.g., 50% (Eh = E0) to 98%.
Table 2.7 shows some redox systems occurring in milk. The standard potential mentioned is not
the only determinant factor because the concentration of each redox system present also
determines to what extent a system of a different standard potential can be oxidized or reduced.
Moreover, the concentration determines the sensitivity of Eh to additions such as oxidants - in
other words, the poising capacity.
Table 2.7 Standard Redox Potentials (E0) of Some Redox Systems Important for Milk, and Their
Concentration in Fresh Milk (T = 25°C)

17
1
Number of electrons transferred per molecule.
2
Probably only partly reversible.
3
In pasteurized milk usually less than 50% of this concentration.
4
In fresh milk irreversible. Action and concentration depend on bacteria.
5
Concentration in the methylene blue reduction test.

In fresh oxygen-free milk, Eh ~ 0.05 V, mainly dependent on ascorbate. On holding the milk,
ascorbic acid shows reactions according to the latter reaction is irreversible, but it only occurs if
both riboflavin and O2 are present; light is active as a catalyst. Riboflavin itself is not susceptible
to O2 but is highly light-sensitive.

In actual practice, however, fresh milk always contains O2 and, accordingly, Eh is higher, i.e.,
+0.2 to +0.3 V. Eventually, only dehydroascorbate is left, which subsequently can be
hydrolyzed. Upon heating of milk, free sulfhydryl groups are formed; these can cause a decrease
of the Eh by about 0.05 V. The cysteine-cystine system itself does not contribute to the Eh
because it is not reversible at neutral pH.
Bacterial action, especially lactic acid fermentation, removes O2 from milk and produces
reductants. Accordingly, the redox potential decreases steeply, ultimately to -0.1 to -0.2 V
dependent on the bacterial species. In this way, methylene blue, if added to milk, is converted to
the colorless reduced form. One takes advantage of this change of color when applying the
methylene blue reduction test for estimating the number of lactose-fermenting bacteria in milk.

18
pH
The pH of milk is higher, or more alkaline, outside of the cow than inside the cow due to loss of
carbon dioxide to the air. The pH of milk is never determined immediately after milking because
the processing milk goes through removes dissolved gasses. The pH is determined after
processing the milk to assure that lactic acid is being produced at the desired rate by added
microorganisms during the preparation of cheeses and fermented milk. The casein in milk forms
into a curd or a gel at a pH of 4.6.

Titratable acidity
Titratable acidity is the amount of alkali required to bring the pH to neutrality. This property of
milk is used to determine bacterial growth during fermentations, such as cheese and yogurt
making, as well as compliance with cleanliness standards. Naturally, there is no lactic acid in
fresh bovine milk, however, lactic acid can be produced by bacterial contamination, but this is
uncommon. The titratable acidity is due to the casein and phosphates.

1.2.3. Changes in composition

The composition of milk may change due to differences in relative rates of synthesis and
secretion of milk components by the mammary gland. Variations are due to differences among
species, between individuals within a strain, and between conditions affecting an individual.
Conditions affecting the cows may include the weather or seasons and the stage of lactation.
Breed:
The US mostly uses milk from cows of the larger breeds, such as Holsteins and Brown Swiss’
because of the lower fat content and greater milk production. Breeds such as the Guernseys and
the Jerseys have higher fat contents in their milks. Both the Guernseys and the Jerseys have a fat
content of 5.2%, where as the Holsteins and the Brown Swiss’ have fat contents of 3.5%.
Diet:
The composition of the cows’ diet and the form in which they are fed affect the composition of
milk and especially milk fat. High fat and/or low roughage diets can reduce the fat content of
milk. Diet has small effects on protein content and none on lactose content. The seasonal effect is
due to the changes in the diet throughout the year.
Stage of lactation
When mammals give birth, their first secreted milk is called colostrum, and it differs greatly in
composition from regular milk. Colostrum contains more mineral salts and protein and less
lactose than normal milk. Also, fat content, calcium, sodium, magnesium, phosphorus, and
chloride are higher in colostrum than in normal milk. Whey content is about 11% in colostrum as
opposed to 0.65% in normal milk.
Colostrum contains an extremely high immunoglobulin (Ig) content. Igs accumulate in the
mammary gland before parturition and transfer immunity to the baby cow. These
immunoglobulins protect the baby cow until it can establish its own Immunity.

19
 CH 2: Microbiology of Milk and Dairy Products
2.1. Bacteria in milk
In addition to being a nutritious food for humans, milk provides a favourable environment for the
growth of micro-organisms. Yeasts, moulds and a broad spectrum of bacteria can grow in milk,
particularly at temperatures between 16 and 35°C. Milk has also a pH of 6.6, which is ideal for
the growth of many organisms. Milk is sterile at secretion in the udder, but it becomes
contaminated by bacteria before it leaves the udder. At this point, the bacteria are few and
harmless, unless the cow has mastitis. Further infection of the milk by microorganisms happens
during milking, storage, handling, and other activities (can enter milk via the cow, air, feeds,
milk handling equipment and the milker). Once micro-organisms get into the milk their numbers
increase rapidly. It is more effective to exclude micro-organisms than to try to control microbial
growth once they have entered the milk. Bacterial types commonly associated with milk are
given in Table 3.1.
Milk producers are required to exclude bacteria from milk, but some may still gain entry. When
the cow has mastitis, microorganisms associated with infections are found, including
Staphylococcus aureus, Streptococcus uberis, and Streptococcus agalactiae. Milk from cows
with mastitis cannot be used for human consumption. Pasteurization destroys most of the
microorganisms in milk and all of the pathogens.

•Pathogenic bacteria: The following pathogenic bacteria are a concern in raw milk:
Mycobacterium tuberculosis, Listeria monocytogenes, Salmonella spp., Bacillus cereus, Yersinia
enterocolitica, Escherichia coli 0157:H7, Campylobacter jejuni
•These pathogenic bacteria can cause foodborne illnesses by ingesting raw milk that contains
these organisms or milk that was not pasteurized properly or was contaminated after processing.
Spoilage microorganisms: Spoilage microorganisms cause degradation of proteins, fats, and
carbohydrates and the milk’s texture, color, taste, and smell is deteriorated. Psychrotrophs
(bacteria that grow at refrigeration temperatures) are usually involved in spoilage reactions. Most
are destroyed by pasteurization, but can still produce heat stable lipolytic and proteolytic
enzymes that can cause spoilage.
•However, some strains and species of Clostridium, Bacillus, Arthrobacter, Cornebacterium,
Microbacterium, Micrococcus, Streptococcus, and Lactobacillus can survive pasteurization and
grow at refrigeration temperatures and cause spoilage.

•Coliforms: Coliforms are closely associated with the presence of pathogens, but they are not
always pathogenic themselves, thus they are called indicator organisms. They are killed by
proper pasteurization treatment. Their presence indicates improper or inadequate pasteurization
or it indicates contamination after pasteurization. Escherichia coli is an example of a coliform.
They are facultative anaerobes and grow best at 37oC or 98.6oF. Coliforms are able to degrade
milk proteins and ferment lactose, causing milk to be spoiled quickly.

•Lactic acid bacteria: Lactic acid bacteria can produce chemical changes that are desirable in the
production of fermented dairy products because they are able to ferment lactose to lactic acid.

20
They are naturally present in the milk and are used as starter cultures in the production of
cultured dairy products, such as yogurt and cottage cheese. Lactic acid bacteria include
lactococci, lactobacilli, and Leuconostoc.

Bacterial types commonly associated with milk are given in Table 3.1.

Milking equipment should be washed thoroughly before and after use - rinsing is not enough.
Microbial growth can be controlled by cooling the milk as most micro-organisms 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 milk temperature will only have fallen to
28°C after three hours. Cooling the milk with cold running water will reduce the temperature to
16°C after one hour. At this temperature bacterial growth will be reduced and enzyme activity
retarded.
Natural souring of milk may be advantageous, e.g. in smallholder buttermaking the acid
developed assists in the extraction of fat during churning. The low pH retards growth of lipolytic

21
and proteolytic bacteria and therefore protects the fat and protein in the milk and it also inhibits
the growth of pathogens. The acidity does not, however, retard the growth of moulds.
Naturally soured milk is used to make many products, e.g. irgo, yoghurt, sour cream, ripened
buttermilk and cheese. These products provide ways of preserving milk and are also pleasant to
eat. 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 directly reflects 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 10 6 cells/ml. High
counts (more than 105 cells/ml) are evidence of poor production hygiene.

2.2. Moulds
Moulds are a heterogeneous group of multicelled organisms which reproduce asexually either by
spore formation or by fragmentation. They can grow on a wide variety of substrates and are
generally regarded as spoilage organisms. However, moulds are used in the production of
antibiotics and in certain cheese varieties. Moulds are aerobic organisms and their growth on
foods can be retarded by excluding air through careful packaging. They can be killed by
relatively mild heat treatments, but mould spores are more resistant to heat.

2.3. Yeasts
Yeasts are unicellular organisms which reproduce asexually by budding. They are used
industrially to ferment carbohydrates to such products as alcohol and citric acid. Yeasts are not
usually used in milk processing and are normally regarded as spoilage organisms in dairy
products.

4.4. Viruses
Viruses are extremely small organisms comprising a spherical head containing the genetic
material, and a cylindrical tail. They must invade other cells to reproduce. Viruses that attack
bacterial cells are known as bacteriophages. Bacteriophages that attack acid-producing bacteria
inhibit acid production in milk thereby causing problems in the manufacture of fermented milks,
yoghurt and cheese.
The most common spoilage micro-organisms of milk and dairy products are Gram-negative rod-
shaped bacteria (e.g. Pseudomonas spp., coliforms), Gram-positive spore-forming bacteria (e.g.
Bacillus spp., Clostridium spp.), lactic acid producing bacteria (e.g. Streptococcus spp.) and
yeasts and moulds.
Milk and milk products are also, to a limited extent, associated with foodborne illness. Disease is
mainly due to consumption of unpasteurized milk containing pathogenic micro-organisms (e.g.
Salmonella, Listeria monocytogenes or Campylobacter).

22