Kendriya Vidyalaya No.

1
Palakkad
Biology Investigatory Project

Pasteurization of Milk

Amna Shirin M
Roll no.: 34
XII A
 Certificate

Certified that the biology investigatory project titled ‘Pasteurization

of Milk’ is a bonafide work of Kumari. Amna Shirin M of class XII
A (academic year 2022-23), Roll no.: 34, Admission no.: 8984. The
project was carried out under the supervision of Smt. Sajeena PGT
Biology.

Teacher in-charge Principal

Practical Examiner Student

 Acknowledgments

I would like to express my gratitude to my teacher Mrs. Sajeena PGT

biology who supported me with patience and gave me the guidance
required to complete this project. This work was an excellent learning
opportunity that helped me learn many new things for which I thank
the CBSE board for assigning this work to us, students.
I would also like to thank my friend for arranging my visit to the
Milma dairy plant which truly helped me gain insight into the
pasteurization methods commonly used and also about the entire
functioning of the plant.
I finally extend my sincere gratitude to all those who have helped and
contributed to my work.
Thank you.
 Pasteurization Of Milk

Definition: According to the International Dairy Federation (IDF):

‘A process applied to a product with the object of minimizing possible
health hazards arising from pathogenic microorganisms associated
with milk by heat treatment, which is consistent with minimal
chemical, physical and sensory changes in the product’.

Pasteurization is the process of heating the product to a predetermined

temperature and
holding it until all or nearly all objectionable microorganisms, which
may be present, are killed.
(This was developed by Louis Pasteur, 1960)

Pasteurization came into use on a commercial scale in the dairy

industry shortly after 1880 in Germany and Denmark.
In general, the term pasteurization as applied to market milk refers to
the process of heating every particle of milk to at least 63°C for 30
min or 72°C for 15s or to any temperature-time combination which is
equally efficient, in properly operated equipment. After
pasteurization, the milk is immediately cooled to 5°C or below.

Objectives of pasteurization
• To make the product safe for human consumption by destroying the
pathogenic
organism, which may be present.
• Improves preservation quality by destroying almost all spoilage
organisms.
• Helps to retain good flavor over a longer period.
Pasteurization requirements for milk- Pasteurization by heating
and time treatments are a compromise among bacterial killing along
with several other factors such as taste, phosphate inactivation, cream
line reduction, etc.
The target microorganism for milk processing is Mycobacterium
tuberculosis (TB germ).

The original method of pasteurization was vat pasteurization, which

heats milk or other liquid ingredients in a large tank for at least 30
minutes. It is now used primarily in the dairy industry to prepare milk
for making starter cultures in the processing of cheese, yogurt,

buttermilk, and pasteurizing some ice cream mixes.

Importance of Pasteurization
• To render milk safe for human consumption by destroying all
the pathogenic microorganisms.
• To improve the keeping quality of milk by killing almost all
spoilage organisms (88-99%)
• Time-Temperature Combination for Specific Requirements.
• All pathogenic organisms are destroyed by pasteurization,
except spore-forming organisms.
• The thermal death point of tuberculosis germs (Mycobacterium
tuberculosis) is slightly higher than that for the inactivation of
the phosphatase enzyme.
Pasteurization was carried out at a heat treatment temperature above
that for phosphatase inactivation and yet below that for cream line
reduction.
 The pasteurization ensures the destruction of pathogens, a negative
alkaline phosphatase test, and the least damage to the cream line.
Encourage slackening of efforts for hygienic milk production and may
mask low-quality milk.

• Diminishes the cream line or cream volume.

•Pasteurized milk- increase the renneting time.
• Fails to destroy bacterial toxins
• In India, pasteurization- is not necessary as milk is invariably boiled
by the consumers.
Pathogenic microorganisms in milk: their source,
hazardous role, and identification
Pasteurization is a very commonly used process of heat treatment of
milk in the dairy industry.
This process kills the harmful pathogenic bacteria naturally found in
raw milk and increases its shelf life. Though it is also the cause of loss
of a few vitamins and minerals.
The duration and amount of heat in pasteurization is based on the
thermal death time of the target microbial population.
In broad terms, pasteurization is grouped into two processes: low
temperature and high temperature.
The low-temperature pasteurization process enables the killing of all
pathogenic microbes.
the high-temperature pasteurization process causes the killing of
vegetative pathogenic and spoilage bacteria. It also causes the
denaturation of serum proteins.
Low-temperature pasteurization includes heating of milk to a high
temperature (63°C for 30 minutes duration or 72°C for 15 seconds
duration) and then immediate cooling (less than 3°C). This process
kills all pathogenic bacteria and reduces the load of spoilage bacteria.
Including this, low-temperature treatment maintains the most
physiological characteristics of milk.
Pasteurization at 72°C for 15 seconds is also called high-temperature
short-time pasteurization or flash pasteurization. The high-
temperature pasteurization process includes the heating of the milk to
high temperature (138°C for 2 seconds, 89°C for 1 second, and 90°C
for 0.5 seconds). Intense heating causes the denaturation of serum
proteins to control syneresis. The shelf life of milk can be increased to

9 months by using ultra-high temperature (UHT) pasteurization. In

this treatment, the temperature of the milk is raised from 135°C–
140°C for 2–4 seconds. This process also targets Coxiella burnetii
which is responsible for Q-fever.
Microbiology of Pasteurized Milk
Pasteurization is intended to make milk and milk products safe by
destroying all the vegetative pathogenic organisms. Pasteurization
systems are designed to provide a 5-log reduction of the microbial
load using the most thermotolerant target pathogen Coxiella burnetii.

With pasteurization, not only are pathogenic microorganisms killed

but also a wide range of spoilage organisms are destroyed.
Raw milk often contains microorganisms at levels of 104–105 CFU,
and the extent to which the number of microorganisms can be reduced
by pasteurization depends not only on the number present initially but
also on the types of organisms.
The spoilage microflora of pasteurized milk is of two types: post-
pasteurization contaminants, which have entered the milk after
heating; and heat-resistant bacteria, which have survived heating. In
general, almost all Gram-negative organisms in milk are destroyed
with pasteurization at 63 °C for 30 min, and although some
thermophilic and mesophilic bacteria, for example, micrococci and
Streptococcus spp., which are thermoduric, may survive
pasteurization, they grow very slowly once the pasteurized milk is
chilled to 4 °C; coryneform bacteria are another group often present in
pasteurized milk, but they grow very slowly in the cooled milk and
rarely cause defects. The methylene blue test is a common quality
control tool for pasteurized milk and decolorization after 30 min
indicates the sufficiency of pasteurization. the usual shelf life of
pasteurized milk should be >4 days under refrigeration.
The endospore-forming genera – such as Bacillus and, to a lesser
extent, Clostridium – can be important in terms of spoilage of
products made from contaminated milk.
Although the anaerobic spore-formers may survive in pasteurized
milk, they usually are unable to multiply owing to high redox
potential; the genus Bacillus, in contrast, is capable of remaining
active after pasteurization,
and its spores may cause spoilage of heat-treated milk. Heat-treated
milk is more suitable for the growth of and enterotoxin production by
S. aureus than raw milk. Therefore, monitoring the presence of this
particular pathogen in heat-treated milk is of paramount importance
concerning the hygienic acceptability of processed liquid milk.

The principal microorganisms growing and causing spoilage of

refrigerated pasteurized milk are psychrotrophic microorganisms, and
as these are heat-labile, the most common origin of psychrotrophic is
post-pasteurization contamination.
There are two major sources of post-pasteurization contamination:
equipment milk residues and aerosols. Thermophilic
microorganisms that survived the heating process can attach to the
surface of plate heat exchangers with high-heat recovery. Growth of
these microorganisms preferentially occurs in a temperature range of
45 to 60 °C in the regeneration section. As a result, the already heated
product is recontaminated before it leaves the pasteurizer. The extent
that bacteria attach to the plates depends on the kind of heat pre-
treatment of the milk before pasteurization. Thermalization of raw
milk or prolonged times of milk circulation in the pasteurizer is the
major factor determining the extent of biofilm formation onto the
heating plates. It is rather difficult to eradicate biofilms on the surface
of milk equipment by applying routine cleaning-in-place protocols.
The filler nozzles, carton-forming mandrels, and pasteurizers are
among the most common sources of post-pasteurization
contamination. The milk contact surface is a route for microbial
aerosols to contaminate pasteurized milk. In particular, airborne yeast,
mold, bacteria, and spores can land on the milk contact surface and
thus contaminate pasteurized milk. The self-enclosed filling units are
much safer than unenclosed filling units in terms of post-
pasteurization contamination of heat-treated milk by airborne
microorganisms.

Bacteria in milk under a microscope

After heating, certain members of the Enterobacteriaceae, including
Serratia, Enterobacter, Citrobacter, and Hafnia, may be numerically
dominant, nevertheless, the ultimate spoilage microflora consists of
psychrotrophic Gram-negative rods, for example, Pseudomonas,
Alcaligenes, and Flavobacterium. Pasteurized milk is required to
satisfy a phosphatase test. Phosphatase is an enzyme that is present in
raw milk indigenously and is destroyed at a temperature only slightly
higher than that used to destroy M. tuberculosis.

In general, flavored pasteurized milk is spoiled faster than

unflavoured pasteurized milk. It was demonstrated that the chocolate
powder used in the production of chocolate-flavored pasteurized milk
stimulated the growth of bacteria in milk, but it did not introduce
additional microbes into the milk. The generation time of bacteria in
flavored pasteurized milk was much faster than in its unflavored
counterpart. In an earlier study, it was found that the growth of L.
monocytogenes in chocolate milk was more pronounced than in skim
and whole milk, and whipping cream.
The fat content of pasteurized milk has no marginal effect on the
growth of pathogenic bacteria. No difference was noted between the
shelf lives of skim (0.1% fat), semi skim (1.6% fat), and whole (3.8%
fat) milk added with or without Pseudomonas spp. at 4 and 7 °C.
Similarly, the numbers of L. monocytogenes in skim milk, whole
milk, and whipping cream did not differ significantly. Therefore, it is
fair to assume that fat standardization has a negligible effect on the
microbiology of pasteurized milk.

The manufacturing technology of concentrated liquid milk includes

pasteurization preheat treatment, evaporation, and cooling. The
condensed milk requires a more intense preheat treatment to ensure
storage stability, a stabilizer may be added, and the finished product is
sterilized in a can by retorting. In general, condensed or evaporated
milk is expected to contain no microorganisms. As a result of
inadequate heat treatment or can leakage, however, the evaporated or
condensed milk may be spoiled. Geobacillus stearothermophilus, an
obligate thermophile, is the organism primarily responsible for the
spoilage mechanism in these products, especially when they are
stored at abnormally high temperatures. Plain condensed milk
products usually contain no additives; therefore, they are stored under
refrigerated conditions. Thermoduric bacteria may survive
pasteurization and heat treatment during evaporation; therefore, high-
quality milk must be used in the manufacture of condensed or
evaporated milk and care must be taken to prevent postprocessing
contamination from the environment and equipment.
Due to the high sugar content and low water activity in sweetened
condensed milk, it is relatively less prone to microbial spoilage than
unsweetened condensed milk. Osmophilic, sucrose-fermenting yeasts
and molds are primarily responsible for the spoilage of sweetened
condensed milk. During filling, the elimination of free air is critical as
molds can grow on the surface of cans when sufficient air is available.
 Visit to Milma Diary plant

As part of the research work, I visited the Milma dairy plant in

Palakkad to gain insight into the pasteurization process followed at a
macro level before the milk packets reach our homes.
The usual method of pasteurization followed in the Milma plant is vat
pasteurization and HTST method.
Long hold or vat pasteurization-
The long hold or vat pasteurization is a batch-type method where the
pasteurization is carried out at 63°C for 30 min.
Types of vat pasteurizers (Classification based on the flow of
heating medium)
1. Spray type
2. Flooded type
3. High velocity flooded type
General requirements
The following are the requirements for a successful batch
pasteurization process.
• Rapid heating: Generally, the circulation of the heating medium
is started as soon as the filling of the vat is begun, thus
shortening the heating time.
• Immediate cooling: In some designs, the cold water is circulated
over the outside of the inner lines as soon as the holding period
is completed, so a part of cooling can be done in the vat itself.
• Heating medium should be only a few degrees warmer than milk
to prevent the formation of milk stones on heating surfaces and
cause minimum injury to the cream line or flavor.
Agitation- Agitation of milk within a certain degree helps in
improving heat transfer.
➢ Agitation is easier in the case of hot fluids than cold ones.
➢ Agitation should not develop foam and it should not injure the
cream line.
➢ The viscosity of the fluid greatly affects the type of agitator.

For vat pasteurizers, an electric or air-operated control can be

connected with a timing clock so that the heat is shut off when the
proper milk temperature has been reached and a bell rings when the
proper length of holding time has elapsed. Also, the temperature of
heating water can be controlled during the holding period
Advantages
• Well suited for small plants, low-volume products
• Variety of products can be handled.
• Well suited for cultured products such as bottled milk, sour cream,
etc.
• Simple controls
• Low installation cost
Disadvantages
• Batch type
• Slow process
• As the controls are mostly manual, it requires constant attention.
• Both heating and cooling are relatively expensive (as we do not have
heat regeneration).

HTST pasteurization-
High-temperature short-time pasteurizers are usually continuous flow
systems using heat exchangers. Generally, plate-type heat exchangers
with regenerative heating, heating, and cooling systems are used.

For HTST pasteurization, the following controls are very important.

• Flow rate
• Temperature
• Pressure

Control for flow rate

• The flow rate is regulated by a metering or timing pump
• A positive displacement pump of rotary or piston type is used.
• Often variable speed drives are used to change the rate of flow when
desired.

Control for temperature

• Control of temperature includes means for maintaining a uniform
product temperature at or above the legal minimum at a safe value.
• It should divert the flow back to the system if the temperature at the
end of the holder is below the legal minimum.
• Usually, a safety thermal limit recording controller is used. (It makes
a continuous record of the temperature and marks the time when the
flow diversion valve operates.)

Control for pressure

Pressure control is important in the following three sections.
• In the regenerators
• In the flow diversion valve
• In the diverted milk lines (for homogenization, etc.)

Advantages of HTST pasteurization

• Uniform treatment.
• Temperature is regulated at close limits and overheating is
prevented.
• Economical than batch systems (due to regenerative heating).

Disadvantages
• The system is complicated.
• Not portable.
• Installation cost is more.