Milk hygiene

Biruhtesfa A.(DVM,Msc12 Associate prof.)

Objectives

At the end of the course, the students will able to:

• Define milk
• Identify the chemical composition and physical properties of
milk
• Identify milk products
• Identify diseases due to milk consumption
• Explain the sanitary practices and hygienic production of milk
• Explain how to make milk safe
• Elaborate the different types of laboratory examination of
milk
outline
• Introduction
• Milk composition
• physico-chemical properties of milk
• Microbiology of Raw and Market Milk
• Quality Control
• Laboratory examination of milk
• Milk-borne diseases
• Lactoperoxidase sytem.
DEFINITIONS

• Adulteration of milk: This is the adding of water or

extracting the fat by removing the cream.
• Milk fat: This refers to the pure fat which consists of
glycerol and fatty acids.
• PH: This is the hydrogen ion concentration of milk.
• Raw Milk: This is a secretion produced by all mammals
before it has been altered to into any other form.
• Cholesterol: This is an alcoholic compound of complex
structure that, in milk, is mostly found in the fat globules.
• Lactation: This is the giving of milk by mammals.
• Specific gravity of milk: This is related to its density. It is
found by dividing the density of the substance by the density
of water at either 40C (39 oF) or 20 oc (68 0F).
• Mammary glands: These are special glands found in all
mammals. In female mammals the mammary glands
produce milk.
• Milk is the first natural food of all young mammals
during the period immediately after birth.
• The public health experts have defined milk as to be
“the lacteal secretion of the mammary
glands of a mammal,
• practically free from cholesterol, obtained by the
complete milking of one or more healthy cows which
contains not less than 8.25% milk solids-not-fat, and
less than 3.25% milk fat.”
 introduction
• Milk contains all the nutrients required by the
neonate and has thus long been recognised as
perhaps nature’s ultimate food.

• furthermore, it is also a rich source of protective

agents,enzymes and growth factors.
• The public health reasons for the terms included in this
definition are as follows: The food value of milk depends
upon its milk fat and milk solids-not-fat content.

• If either of these contents is reduced to below

the range for normal market milk, the proteins,
carbohydrates, minerals and certain vitamins are also
reduced.
Practical experience shows that 3.25% of milk fat and
8.25% of solids-not-fat are a reasonable minimum for
mixed herd milk.
 Chemical composition of Milk
Milk composition
• Milk is often described as a colloidal suspension,
containing emulsified globules of fat,a heterogeneous
family of major and minor proteins, the carbohydrate
lactose, minerals,vitamins and enzymes.

• It is the most nearly perfect food. It is not ‘the’

perfect food because it is not an entirely dependable
source of all vitamins (vitamins C and D) nor does it
contain sufficient iron.
1. Water

• This is the principal constituent of milk and is the

medium in which all constituents are in solution
or in suspension.

• In some countries, such as Ethiopia, where the

production and distribution of milk is not
properly controlled, the adulteration of milk by
the addition of water is commonly practiced.
 [Link]

• Proteins are built up of amino acids in various combinations.

• The three principal proteins found in milk in colloidal

suspension are casein, lacto albumin and lacto globulin.

• A complete protein contains all the essential amino acids and,

therefore, has a high nutritive value, especially suited to the
growth requirements of children.

• Casein is a complete protein, insoluble in water and is found

only in milk. It exists in milk in combination with calcium
phosphate and this combination causes precipitation.
• The acids in milk take the calcium
from calcium phosphate and thus throw the insoluble casein out
of the solution as curd.

In the normal curding or souring of milk the casein is precipitated

by lactic acid produced through the action of bacteria upon
lactose.
3. Lactose (Milk Sugar)

• Lactose is composed of the simple sugar glucose and galactose.

• Lactose, like glucose, is a reducing sugar. Lactic acid bacteria
present in milk readily germinates it. These bacteria produce an
enzyme known as lactase which splits the lactose and ferments it
into lactic acid and other products.

• This causes the milk to sour. Lactose by nature gives milk a sweet
flavor and furnishes the body with heat and energy.
• Variations in the fat content are influenced by factors such as:
• Breed of cow: Breeds producing large amounts of milk, such
as Holstein, secrete milk of a lower fat content.
Locally there are breeds which produce a low amount of milk
but have a high fat content.
Age and health of cow: Fat content decreases due to old age
and poor health.
Stage of lactation: Fat content decreases during the first 2 or 3
months of lactation.
• Season of the year: Fat content is lowest in late spring or
early summer.
• Feeding habit: The fat content of milk varies depending on the
type of fodder that the cows use.
• Variation during milking:
a. The “fore milk” or “first milk” drawn is lowest or poorest in
fat content.
b. The “middle portion” is average in fat content.
c. The “stripping “or “last milk” drawn is highest or best in fat
content.
Minerals

• The major inorganic salts present in milk are calcium,

phosphorous, sodium, potassium, magnesium chlorides and
sulphates.
• Trace elements such as iron, copper, manganese, iodine and
zinc are also present in milk.

• Milk is a source of calcium and a rich source of phosphorous.

Both of these elements assist in the formation of bones and
teeth of growing children. Since milk is deficient in iron, an
exclusive milk diet may cause anaemia due to iron deficiency.
Vitamins
• Milk may be rated as a good source of many vitamins in
certain variation. The most common vitamins present in milk
are:
• Vitamin A: Milk is rich in, and the best source of, vitamin A,
which is heat resistant
• Vitamin B1 (thiamine): Milk is a fairly good source of vitamin
B1, which is heat resistant
• Vitamin C: Milk is a less dependable source of vitamin C,
which is heat sensitive
• • Vitamin D: Milk is a relatively poor source of vitamin D,
which is heat resistant
• • Vitamin E: Milk is a fairly good source of vitamin E, which is
heat resistant
• • Vitamin B2 (Riboflavin): Milk is a rich and outstanding source
of vitamin B2, which is heat resistant .
• The fat of milk is a rich source of vitamin A. Therefore butter,
cream and all kinds of cheese products which are made from
whole milk are outstanding sources of vitamin A.
• Naturally the amount of vitamin C content is very minimal. It is
also easily affected by heat treatment.
• Brief boiling of milk, as it is commonly practiced in most local
homes, results in the loss of 20-50% of its vitamin C content.
Enzymes
• Enzymes are protein compounds which act as biological
catalysts having the ability to speed up or retard the chemical
reaction without themselves being changed.
• Enzymes are specific in their action in
that they act by splitting certain specific substances. For
example,
• lipase acts only on fat, lactase acts only on lactose, phosphates
acts only on phosphate etc.
• Enzymes are relatively [Link] temperatures, an
unfavorable pH, light, and certain chemicals tend to destroy or
inactivate them.
• The principal enzymes present in milk are:
• Phosphate: Acts on phosphates. It is a heat liable enzyme and
is a normal constituent of raw milk. It is also important in
connection with tests for the determination of proper
pasteurization as it is almost completely inactivated at normal
pasteurization temperatures.
• Lipase: Acts on fat or lipid.
• Lactase: Acts on lactose (milk, sugar).
• Others like amylase, galactase, catalase, etc
Gases

• The principal gases in milk are CO2, O2and N2. They are
introduced
into the milk during the process of milking and handling.
• Lactose is a sugar found only in milk. It is also present in dairy
products and products made from milk, including cheese and
ice cream.
• If a person has lactose intolerance, their digestive system
produces too little of an enzyme known as lactase. Lactase is
needed to break down lactose.
• Lactose intolerance is different from a milk allergy. In a milk
allergy, the body reacts to milk proteins, not milk sugar. A milk
allergy can result in severe symptoms, including anaphylaxis.

• Worldwide, an estimated 68% of people have trouble

digesting lactose. The figure is lower in the U.S., affecting 36%
of individuals.

• People with lactose intolerance experience symptoms after

eating or drinking milk or dairy products that contain lactose.
• Symptoms can range from mild discomfort to a severe
reaction. This depends on how much lactase a person’s body
produces and how much lactose they consumed.

• Most people with lactose intolerance can eat some amount of

lactose without experiencing symptoms. Each person has a
different tolerance level
Lactose tolerance test

• In a lactose tolerance test, you'll be given a drink of lactose

solution and a blood sample will be taken.

• If you're lactose intolerant, your blood sugar levels will either

rise slowly or not at all.

• This is because your body is unable to break down the lactose

into glucose.
Milk tolerance test

• In a milk tolerance test, you'll be given a glass of milk (about

500ml) and your blood sugar levels will be tested.

• If your blood sugar levels do not rise after drinking the milk,
you may be lactose intolerant.
Hydrogen breath test

• A hydrogen breath test is a simple way of determining if you

may be lactose intolerant.
• You'll be asked to avoid eating or drinking during the night
before the test.
• When you arrive for the test, you'll be asked to blow up a
balloon-like bag.
• This sample of your breath will be tested to find out how
much hydrogen is present, measured in parts per million
(ppm).
• You'll then be given a drink of lactose solution and your breath
will be tested every 15 minutes over the next few hours to see
if the level of hydrogen changes.
• If your breath contains a large amount of hydrogen (more than
20ppm above your baseline) after consuming the lactose
solution, it's likely that you're lactose intolerant.
• This is because lactose intolerance can cause the bacteria in
the colon (large intestine) to produce more hydrogen than
normal.
Physisco chemical property

• In milk from most mammals, lactose is the major

carbohydrate, but small amounts of other carbohydrates also
occur; cow’s milk contains∼10 mg L−1 monosaccharides
(glucose and galactose) and ∼100 mg L−1 oligosaccharides.
• Lactose is a disaccharide composed of d-glucose and d-
galactose; the aldehyde group of galactose is linked to glucose,
through a -1,4-glycosylic linkage
• Lactose is one of the many constituents in milk that undergo a
variety of changes on heat treatment

• The primary salts in milk are phosphates, citrates, chlorides,

sulphates, carbonates and bicarbonates of sodium, potassium,
calcium and magnesium
• The average mineral composition of bovine milk is given in
Table
• Since almost all the lipids in milk are found in milk fat globules, milk can
be considered as an oil-in-water emulsion, the physico-chemical aspects
of which are essential when considering changes that occur on storage
or processing of milk.
• Cow’s milk generally contains 30–35 g L−1 protein which is commonly
divided into two classes on the basis of the solubility at pH 4.6:
i-the insoluble caseins, which represent ∼80% of total milk protein, and
ii. the soluble whey (or serum) proteins, which represent
∼20% of total milk protein
• In cow’s milk, ∼20 enzymes have been characterised. Furthermore,
the presence of another 40 enzymes has been demonstrated via their
activity.
Plasmin,lactase
Lipoprotein lipase
Alkaline phosphatase
Lactoperoxidase*
Milk may be rated as a good source of many vitamins in certain
variation.
The most common vitamins present in milk are:
• Vitamin A: Milk is rich in, and the best source of, vitamin A,which is
heat resistant
Vitamin B1 (thiamine): Milk is a fairly good source of vitamin
B1, which is heat resistant
• Vitamin C: Milk is a less dependable source of vitamin C,
which is heat sensitive.

• Vitamin D: Milk is a relatively poor source of vitamin D, which is heat

resistant.
• Vitamin E: Milk is a fairly good source of vitamin E, which is heat
resistant.
• Vitamin B2 (Riboflavin): Milk is a rich and outstanding source of vitamin
B2, which is heat resistant.
 1. Physico-chemical properties of milk

2. Real case story: Brazil

Brazil Govt was interested to promote goat milk
production, and giving some subsidy unfortunately
some farmers are brave enough that they were mixing
cow milk with goat milk and sell for the govt.
What do you advice the Brazillian govt to
differentiate/screen this type adulteration. goat milk
with cow milk
physico-chemical properties of milk

• PH

• Organoleptic quality

• Like colour, smell, taste,

• Boiling pt

• Freezing point
 physico-chemical properties of milk

• The pH of normal milk from a healthy cow is in the

range 6.6–6.7;
• lower pH values would suggest growth of
microorganisms that ferment lactose to lactic acid
or extensive lipolysis
• higher pH values in milk may be encountered in
times of physiological stress for the producing
animal, when the mineral balance of milk is altered
by changes in the permeability of the blood–milk
barrier
• Color: The milk pigments influencing the color of milk are:•
Carotene (Vitamin A) of the fat in the milk that gives a golden color.

Specific gravity
• Milk contains many constituents whose specific gravity is greater
than that of water. Therefore milk is heavier than water.
• Fat is the only important constituent with a specific gravity of less
than 1. (1.027–1.035 g/ml)
• Hence the more fat present in the milk, the lower the specific
gravity of milk.
• With the removal of fat, as in skimmed milk, the specific gravity is
increased
• Riboflavin (B2) of the whey that gives a bluish color.
Freezing point:
• Pure water freezes at 00 c (320 F). Since the freezing
point of a dissolved substance is lower than the freezing
point of a solution, milk freezes at -0.550c (31.010 F).

• Boiling point : Water boils at 1000 c (2120 F) while milk

boils at 100.170 C (212.30 F) at sea level.
 Adulteration of milk and milk
products
• Adulteration of milk: This is the adding of water
or extracting the fat by removing the cream.

• What is Milk safety and Milk quality?

Adulterant means any material which is or
could be employed for making the food unsafe
or sub-standard or misbranded or containing
extraneous matter.
• Adulturated food is dangerous because it may be
toxic and can affect health it could deprive
nutrients essential for proper growth and
development.

• Table (tests for adulturations

 Microbiology of Raw and Market Milks

• Milk is an ideal medium for the growth of many organisms,

having a high water content and abundant nutrients, and
being nearly neutral pH (6.4–6.8).

• A plentiful supply of food for energy is available in the form

of milk sugar (lactose), milk fat, citrate, and nitrogenous
compounds (proteins, amino acids, ammonia, urea and
other non-protein nitrogenous compounds).
• Microorganisms present in milk can be classified
into two main groups: pathogenic and spoilage
organisms, although some may play a dual role
(e.g. Bacillus cereus).

-In addition, like lactic acid bacteria.

• Pathogenic organisms are those capable of inducing food
poisoning, thus posing a threat to public health.

• By virtue of their elaborated enzymes (e.g. protease,

peptidase, lipase, esterase, oxidase, polymerase, -
galactosidase), spoilage organisms are capable of
hydrolysing milk components such as protein, fat and
lactose in order to yield compounds suitable for their growth.

• Such reactions can lead to spoilage of milk, manifested as

off-flavours and odours,and changes in texture and
appearance
 Spoilage organisms
Gram-negative psychrotrophic bacteria
• The growth of psychrotrophic bacteria is of major concern
when raw milk is kept at low temperature
• During growth of these bacteria, heat-stable enzymes such as
proteases and lipases are formed and consequently cause
protein and lipid breakdown and related defects
Pseudomonas spp.
Enterobacteriaceae
others like Flavobacterium, Achromobacter,
• Aeromonas, Alcaligenes and Chromobacterium
 Spore-forming bacteria
• Bacillus spp is a predominantly
B. stearothermophilus
B. cereus
Clostridium
Lactic acid bacteria
 Pathogenic organisms
• The prevalence of these varies considerably, depending
on geographical area, season, farm size, number of
animals on farm, hygiene and farm management practices

• Campylobacter jejuni, Shiga toxin-producing Escherichia

coli, S. aureus,Listeria monocytogenes, Salmonella spp.
and Yersinia enterocolitica
 Pathogenic organisms in raw milk
are of two types
Those involved in bovine mastitis and those that
externally contaminate milk.
• The bacteria causing mastitis include S.
aureus, Streptococcus agalactiae, Str.
dysgalactiae, Str. uberis, Listeria spp. and E.
coli.
• Salmonellae and thermoduric Campylobacter
strains are the most common pathogenic
bacteria from sources external to the udder
 Milk-borne diseases
Milk is an efficient vehicle for a great variety of disease
agents: The sources of infection or contamination of
milk may be
[Link] dairy animal
2. human handler or
3. the environment, e.g., contaminated vessels, polluted
water, flies, dust, etc.

A joint FAO/WHO Expert Committee on milk hygiene

classified milk -born diseases:
1. Infections of animals that can be transmitted to
man:
Primary importance –
Tuberculosis
Brucellosis
Streptococcal infections
Staphylococcal enterotoxin poisoning
Salmonellosis
Q Fever
• Lesser importance –
Cowpox
Foot and mouth disease
Anthrax
Leptospirosis
Tick-borne encephalitis
Infections primary to man that can be transmitted
through milk:
Typhoid and paratyphoid fevers
Shigellosis
Cholera
Enteropathogenic Escherichi coli
Non-diarrhoeal diseases
Streptococcal infections
Staphylococcal food poisoning
Diphtheria
Tuberculosis
Enteroviruses
Viral hepatitis
 Milk products
• Some of the common milk products that are made locally or
commercially from whole milk are

• Cream

• A portion of milk containing not less than 18% milk fat.

• Cream may be taken from milk by “skimming “or “separating”
Skimming is the process of removing manually the cream,
which rises to the surface, after milk stands in a container. The
remaining part of the milk is called skimmed milk. Separating
is the process of removing cream mechanically.
• Curd
• The coagulated part of milk. If milk is stands in a container for
sometime at room temperature, it forms clots called curds
which are contained in a clear liquid called whey.
• Whey:
• This is the watery part of milk after separation of the curd from
the whole milk. It contains protein, lactose, minerals and salts.
• Cheese
• This is the clotting casein of milk. Cheese is made from
separated milk or whole milk. The milk curd, after being
removed from the whey, is pressed into solids and through
other processes and forms cheese. Genuine cheese must
contain no fat other than that obtained from milk.
• Butter

• This is the solidified milk fat or cream prepared by churning.

The cream produced from milk is violently churned up and
shacken so that the fat globules are broken up and closed
together into pieces of mass called butter
 Sanitary practices to be
observed in producing safe milk
• The main objective is to protect milk from external
contamination such as animals, the milker and the
environment in which the milking is done.
Sources of contamination : Preventive Measures
Contamination from human discharge and wastes:
• Construction of a proper waste disposal system
• Prevention of human discharge and wastes from
contacting animals and milk.
• Practices of good personal hygiene.
• Segregation of animals from human habitation.
Contamination from air borne dust and droplets:
• Proper construction of the milking area
• Sneezing or coughing away from milking containers
during milking
• Provision of clean surroundings and avoidance of
dusty conditions during milking
Contamination from animal bodies, hides, udder and
teats:
• Clipping, brushing, cleansing and sanitizing before
milking
Milk containers (contamination from milking utensils
and dirty water):
• Proper washing and storing of milking containers
• Using proper and easily cleanable utensils
• Using only safe water for washing and cleaning
Milk handlers and contamination from milkers and
their clothing:
• Keeping the milker’s level of personal hygiene high
• Washing of hands with detergents before milking
• The milker should always wear clean garments while
milking.
Cow and contamination from diseased animals:
• Detecting and isolating affected animals
• Veterinary inspection and supervision of animals
• Treatment and vaccination of animals.
Steps of milking milk by hand:

• Prepare all milking materials prior to milking

• Massage and wash teats and udder thoroughly with cloth or piece of
towel dipped in warm water; if possible in warm bactericidal (chlorine)
solution
• Draw out foremilk in strip cup squeezing a few hand squirts
from each teat
• Fore milking stimulates milk flow.
• Fore milking helps detect an abnormal milk resulting from a diseased or
injured udder.
• Before starting milking be sure that milker’s hands are clean and dry
• Milking is done carefully without letting any foreign objects or vermin
enter the open milking jar.
• After finishing milking transfer the milk into a clean container by
straining and immediately keep or store the container at a temperature
of 500 F (100 C) or less.
 Methods of making milk safe
Milk as secreted by the udder cells of a healthy cow is probably
sterile, i.e. it contains no micro-organisms capable either of
souring the milk or causing disease.

But when milk reaches its reservoir within the udder, and
particularly in passing through the teat of the udder and
reaching the milking pail, the risk of picking up deterious
microorganisms steadily increases.

• There are at least five methods of treating milk.

 i. Boiling
• Boiling of milk destroys all microorganisms except
the spore formers but it changes the nutritive value
of milk, its flavors and palatability and appearance.

• However this disadvantage should be disregarded in

favor of the safety of boiled milk i.e. its freedom from
disease-causing microorganisms.

• Boiling is technically difficult to process on a large

scale and is commercially uneconomical.
 ii. Pasteurization
• The term pasteurization, named after Louis Pasture (1822-
1895) the famous bacteriologist, was first devised to be
applied to other fluids, the main one being milk and used
for controlling spoilage in wine and beer. Afterwards
pasteurization became popular.

• Pasteurization of milk is a universally known method of

rendering raw milk safe through controlled heat treatment

• Mycobacterium tuberculosis

• coxiella burnetii (Rickettsia burnetii)

 Pasteurization

• defined as the process of heating every particle of

milk and milk products to a predetermined
temperature and holding this temperature for a
predetermined time.

• Pasteurization of milk is not sterilization but it is a

destruction of all pathogenic micro-organisms,
a good number of other non-pathogenic and
non- spore forming bacteria and certain enzymes
in the nutritive value and the chemical nature of the
milk.

• 3 methods of pasteurization
The holding or vat method
• The holding or vat method, also known as the low
temperature holding time process, is a method of holding the
milk in a vat (container) to a temperature of 630c (1450
• F) for 30 minutes.

• In this process milk is heated in a large jacketed container by

steam or hot water circulating in the interspaces of the vat or
through a pipe coil in the vat to a temperature of 1450 F.
• The high temperature-short time method
• is a continuous process by which milk is rapidly brought to a temperature of
710c (1610F) and heated continuously for 15 seconds.

For all practical purposes if milk is said to be properly pasteurized it

must satisfy the following three conditions:

• The milk must be cooled adequately (500F or less) before past

eurization so as to prevent the formation of heat resistant
staphylococcal entrotoxin
• The pasteurization equipment should function properly and
adequately
• Precautions should be taken to prevent any post pasteurization
contamination.
The ultra-high temperature (UHT) method
• In this process the milk is heated to at least 880 C
(1910F), for at least one second and then
immediately cooled to at least below 100C (500F).

• developed very recently and is used only in a few

developed countries because it requires complex
equipment and the highest levels of precision and
handling.
 iii. Sterilization

• In this process milk is heated to destroy all micro-

organisms including spore forming and can only be
done by keeping the milk at a temperature above
normal boiling point (1000 c or 212 0 F) for at least 20’

• If the T0 is higher and the sterilization effect is greater,

>more marked change in the color and
taste of the milk.

• The nutritive value of the milk, (heat sensitive

vitamins) and proteins, is affected depending upon the
type of the sterilization process.
 iv. Drying

• A method where the entire water constituent is

removed from the milk by evaporation. The solids
remaining form what is called milk powder (dry milk).
Milk powder can then be made into liquid milk by
adding a proper amount of water.

• Drying is not a simple operation that can be

accomplished by heating and evaporation in the
home. It needs special equipment and arrangement.

• There are two known methods of drying.

• Roller drying

• The milk in small quantities is slowly poured over a heated and

revolving roller. The roller is heated by steam or hot air. The heat
in the roller will evaporate the water constituent of the milk. The
solid parts will remain behind which will be collected and
packaged.
• Spray drying
• This is also a mechanism whereby the milk is subjected to hot air.
The process is simply spraying the milk through a current of hot
air. The remaining powder will also be collected and packaged. In
both processes the milk powder is collected in sterile containers,
usually cans. The cans are sealed airtight under sterile conditions,
labeled and distributed to consumers in near and far away places
 v. Cooling

• Since milk is an ideal medium for the multiplication

of most disease producing organisms, the milk
should be kept at 500F or below starting immediately
after milking.

• This is very important especially if milk is going to

be delivered raw to consumers. This temperature
range is essentially needed to arrest the growth of
micro-organisms specially brucella organisms.
However milk should not be allowed to freeze.
The simplest method of cooling milk is submerging
the cans of milk in troughs or tanks of ice water if
possible. A trough should be concrete, preferably
insulated by layers of cork or cotton pads. A 120-liter
tab of water kept at 370F will cool 40-liters milk from
850F to 450 F, the recommended temperature.

------------------/--------------------------/-------------------
Lactoperoxidase system in milk

• antimicrobial substances present in bovine milk are

lactoferrin,lysozyme,lactoperoxide,and lactoglobulins.
• Lactoperoxidase (LP), a hemoprotein present in milk,
tears, and saliva
• Lactoperoxidase has no antibacterial effect on its own but
has the ability to oxidise the thiocyanate ion (SCN-) in the
presence of hydrogen peroxide (H2O2)
• The Lactoperoxidase-thiocyanate-hydrogen peroxide
interaction constitutes what is referred to as the LP
sysytem, where in hydrogen peroxide serves as a
substrate for LP in oxidizing thiocyanate (SCN-) and
iodide ions, resulting in the generation of highly
reactive oxidizig agents.
 Lactoperoxidase
SCN - + H2O2 ------------------------ OSCN - + H2O
(Thiocyanate ion) + (Hydrogen Peroxide)------ (Antibacterial compound)

The method of reactivation of the LP-s in milk is to add

about 10 parts per million (PPM) of thiocyanate (preferably
in liquid form) to the raw milk to increase the overall level
to 15 PPM (circa 5 PPM naturally present).
The solution is thoroughly mixed for 30 seconds and then
an equimolar amount (8.5 PPM) of hydrogen peroxide is
added (generally in the form of a granulated sodium
carbonate peroxyhydrate).
• This treatment reactivates the antibacterial effect of the
LP-s in the raw milk and extends the shelf life of raw milk
under tropical conditions for a further 7 to 8 hrs. This
means that producers can transport their milk to the
collection point where it is treated with LP-s and can
then be transported by whatever means are available
• The LP sysytem has the ability to inhibit
bacteria,fungi,parasites, and viruses and thus is
considered a broad-spectrum natural antimicrobial
contributing to protecting the gut of weaning calves from
enteric pathogens, protecting the mammary gland from
disease,and ideed preserving milk.

Occurrence and biosyntesis

• LP is synthesized and secreted by ductal epithelial cells

of the mammary gland and other exocrine glands
• -The level of LP in bovine milk is about 20 times
larger than that of human milk and changes
constantly during the postpartum period.

• Thiocyanate,which is required for the antimicrobial

activity of the LP system,may be present in
significant amounts in milk,where as hydrogen
peroxide may be generated by microbial
flora,usually bacteria in mammary gland.
• Hydrogen peroxide(H2O2) is the third component of
the LP system. Many lactobacilli,lactococci, and
streptococci produce suffiecient H2O2 under aerobic
conditions to activate the LP system.

• In bovine milk, the intial concentration of LP in

colostrum is low, increasing to a peak at 4 to 5 days
postpartum,after which it declines to a level
considered relatively high and remains unchanged
at that level during lactation.
Antimicrobial activity

The structural damage to microbial cytoplasmic

membranes through oxidation of SH groups causes
leakage of potassium ions,aminoacids and peptides
into the medium as well as inhibition of the uptake of
glucose,aminoacids,purines, pyrimidines and
subsequent sythesis of proteins,RNA and DNA.
• Different groups of bacteria show a varying degree
of sensitivity to the LP system.
• Gram negative,catalase ‘+’ organisms, such as
pseudomonas, ecoli/coliforms, salmonellae,shigellae
are not only inhibited by the LP system but also,
depending on the medium
conditions(PH,temperature,incubation time) may be
killed.
• Gram- positive,catalase negative bacteria,such as
streptococci and lactobacilli are generally inhibited
but not killed by the LP system.
 Quality Control
• Quality control has the objective of ensuring the
quality and safety of the milk offered to the
consumer
• The best way to manufacture milk with good quality
is to start with a good raw material.
• Testing raw milk is thus essential to ensure safety and
quality.
• Raw milk is analysed for the presence of macroscopic
abnormalities, addition of water, microbial quality, presence of
milk from mastitic cows, presence of residues, and composition.

• Microbial contamination is a major issue, since pathogens can

compromise the safety and spoilage microorganisms can limit the
shelf life.

• The European Union (EU) establishes limits for total bacterial

counts (≤50 000 colony-forming units (cfu)mL−1), S. aureus and
Salmonella spp. Other pathogens or their toxins should not be
present in amounts that can become a health hazard.
• Methods of analysis
Analysis of abnormal milk
Microbial analysis
Mastitic milk – somatic cell count (SCC)
Testing for residues compounds
1, Analysis of abnormal milk
• Abnormal milk, containing clots or blood, should not reach the bulk
tank. To avoid it, milk can be tested visually after filtration to observe
clots.
2, Microbial analysis
• Microbial tests are applied to detect poor
hygiene conditions and to evaluate the microbial
quality of raw and processed milk.
• Total bacterial load is used to get an overview of the
microbial contamination; psychrotrophs are assessed to
prevent potential spoilage, coliforms to evaluate the
hygiene history of milk.

• specific pathogens (e.g. Salmonella spp., S. aureus,

Listeria monocytogenes, Mycobacterium
paratuberculosis) to prevent potential health hazards and
thermoduric bacteria to assess the hygiene of the
production system.
• Physical methods such as conductivity and titratable
acidity are indirect but rapid sensing methods to
estimate the microbial content. Measuring the pH is
a routine practice in milk testing. These methods
lack sensitivity and specificity.

• Counting methods have been the most popular

methods. The standard plate count (SPC) has been
working successfully for a long time and is the
reference method used.
• Immunodetection methods are based on the ability
of antibodies to bind specific proteins on bacteria.
They are useful to detect pathogens.

• DNA-based methods are all based on the detection

of specific DNA sequences. Techniques based on
nucleic acid hybridisation and polymerase chain
reaction (PCR) are useful to detect specific
pathogens or groups of microorganisms.
3,Mastitic milk – somatic cell count (SCC)
• Mastitic milk contains pathogens and spoilage
microorganisms, and it is characterised by an increase in
Na+ and Cl− as well as leucocytes.
• Screening tests include conductivity tests,based on
changes in the ion composition, and the California
mastitis test (CMT), based on the increase in leucocytes.
• More accurate methods are based on the enumeration
of somatic cells, mostly leucocytes that should not
exceed 400 000 or 750 000 cells mL−1 (EU and US milk
quality standards, respectively).
4,Testing for residues compounds
Milk contaminated with residues in levels that are
potentially harmful to human health should not reach the
tank. The major analytical challenge for the detection of
residues is the large number of compounds and the low
amounts to be detected.

• Antibiotics comprise a considerable number of

compounds, including –lactams.
(penicillins),sulphonamides, tetracyclines,
aminoglycosides and macrolides.
• Pesticides comprise a great variety of groups that
include different chemical species.

• Two groups of insecticides, organophosphates and

carbamates are widely used

• Mycotoxins, particularly aflatoxin M1, are secreted

into milk of dairy cows

• The more reliable analytical method includes an

immunoaffinity clean-up step, followed by high-
performance liquid chromatography (HPLC) and
fluorimetric detection. ======/========
Laboratory examination of milk
• is one of the most valuable and necessary aids to
the overall milk quality control program.
1, Sediment Test : to determine the amount of
insoluble visible filth or extraneous matter in the
milk.

2, Methylene-Blue Reduction Test: to determine the

quality of milk before pasteurization.
- Involves determination of time required for the
disappearance of color when methylene blue
thiocynate solution is added to raw milk.
3, Resazurin Reduction Test: to determine the
sanitary quality of raw milk.
4,Phosphate Test : to determine the efficiency of
pasteurization or to check the adequacy of
pasteurization.
5, Babcock Test (Fat Test) : to determine the fat
content of milk.
6, Coliform Test : to determine the presence of
organisms of coli form group in milk.
7, Standard plate count (agar plat count) : to
estimate the bacterial population of the milk and
dairy products prior to heat treatment.
8, Clot-on-boiling test
9,Organoleptic test sense organs
10, Alcohol test
11, Lactometer test - specific gravity