Dairy starter cultures

Dairy starter cultures are harmless microorganisms of known and stable metabolic activities that
are inoculated into milk to produce fermented products of desirable appearance, body, texture
and flavor.
When inoculated in milk they convert the milk sugar, lactose, into lactic acid for curd formation
and other by-products responsible for flavour, aroma and consistency development.
Starter cultures are used in the manufacture of yoghurt, kefir and other cultured milk products as
well as in butter making and cheese making.

Classification of starter cultures

Lactic acid bacteria are the prime agents in producing soured (fermented) milk and dairy
products. Although they are genetically diverse, common characteristics of this group of
bacteria include Gram-positive, non-motile, and non-spore forming. They grow anaerobically but
are aero-tolerant.

Taxonomy
There are currently 12 genera of lactic acid bacteria, of which 4 contain organisms used in dairy
fermentations: Lactoccocus, Leuconostoc, Streptococcus, and Lactobacillus. A fifth genus,
Enterococcus, is occasionally found in undefined starter cultures.

N/B Propionibacterium shermanii and Bifidobacterium spp. which are not lactic acid bacteria
are used. In addition, other bacteria including Brevibacterium linens, responsible for the flavor of
Limburger cheese; and moulds (Penicillium species) are used in the manufacture of Camembert,
Roquefort and Stilton cheeses.

A. Classification based on strains

i) Single strain- Every starter consists of a pure culture of one strain. The only problem is
there may be sudden failure of starter due to bacteriophage attack which leads to heavy
loss to the industry.

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  Pure defined cultures are single strain cultures selected from natural mixed
populations for specific properties
 May be rotated to avoid phage infection
 Have the advantages of uniform rate of acid development and uniform flavor profiles

ii) Paired compatible strain: Two strains of cultures having complementary activities in
known proportion are used. This will reduce chances of culture failures. In case of
bacteriophage attack, only one type of organism will be affected and the other organism
will carry out the fermentation without any problem

iii) Mixed Strain: More than two culture organisms which may have different
characteristics like, acid production, and flavor production, slime production etc. in
unknown proportion are used.

iv) Multiple mixed strain: More than two strains in known proportion are used. The quality
and behaviour of these strains is predictable.

B. Classification based on temperature

i) Mesophilic culture
The optimum growth temperature of these cultures is 30 oC and they have a growth
temperature range of 20- 35 oC
Incubation is usually for 14–16 h or until a pH of 5 is reached. If pH control is not used,
the final pH should be 4.8
Examples of Mesophilic cultures
 Lactococcus lactis subsp. cremoris
 L. delbrueckii subsp. lactis
 L. lactis subsp. lactis biovar diacetylactis
 Leuconostoc mesenteroides subsp. cremoris

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ii) Thermophilic culture
Thermophilic cultures grow from 30 to 46°C for 8–10 hrs. A final pH value as low as
4.7 is acceptable.
Thermophilic starters almost always consist of two organisms, Streptococcus
thermophilus and either Lactobacillus helveticus, Lb. delbrueckii ssp. lactis, or Lb.
delbrueckii ssp. bulgaricus.
Examples of Thermophilic cultures
 Streptococcus salivarius subsp. thermophilus (S.thermophilus)
 Lactobacillus delbrueckii subsp. bulgaricus
 L. delbrueckii subsp. lactis
 L. casei
 L. helveticus
 L. plantarum

C. Classification based on forms

i) Liquid culture: for propagation of mother culture which is nowadays very rare

ii) Deep-frozen culture: This is concentrated culture for preparation of bulk starter.
Example
Deep frozen REDI SET - freeze below -45 min 12 months

iii) Freeze-dried culture: This is the concentrated culture in powder form.

Examples
DRI – VAC - fridge below +5 - min 12 months
Freeze dried (DVS) - freeze below minus 18 (-18) minimum storage period 12 months
Transported at room temp (20oC) to a max 10 days then cooled to freezer below -18
Freeze dried REDI SET - freeze below -18 for a minimum of 12 months

iv) Deep-frozen which is the super concentrated culture in readily soluble form, for direct
inoculation of product.
Examples

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 Deep frozen (DVS) - freezer below -45 - minimum storage 12 months
Transported at -100 C to a max 2 days then cooled to freezer below -45

D. Classification based on administration

Starter cultures can be divided basically into two groups:
i) DVS – these are direct-to-vat or direct-vat-set (DVS) cultures, which are inoculated with
the DVI (direct vat inoculation) technique.
DVI involves inoculating the yogurt mix directly with a very large number of frozen or
freeze-dried starter organisms. Although a slightly longer incubation time is required with
the DVI technique, resistance to phage attack will be achieved.
ii) Propagated cultures - the cultures that require preliminary preparation steps before
inoculation. For propagation of mother culture (nowadays fairly rare).

E. Classification Based on the flavour production

i) B ( L) type : contains Leuconostocs as flavour producer ( old name is betacocccus)

ii) D type : contains L. lactis ssp. lactis biovar diacetylactis for flavor production

iii) BD(LD) type : Mixer of both of the above cultures

iv) N or O type : Absence of flavour producing organism

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Qualities/Characteristics considered in selection of Starter Culture

a) Acidity
Starters are selected depending on:
 Maximum acidity development
 Rapidity of acidity development

b) Viscosity
Starters are selected depending on:
 Ability to grow on high total solids

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  Ability to increase viscosity- A number of bacteria produce a mucus (or slime) of
polysaccharides, which dramatically increase viscosity, as they are highly water-
soluble and dissolve in the medium. This is utilized in certain cultured products
such as yoghurt

c) Optimum temperature of incubation

Starters are selected depending on:
 Their optimum growth temperature; Thermophilic for yoghurt and yoghurt
products and mesophilic for cultured milk products

d) Salt tolerance
Salt can slow the growth of bacteria

e) Production of typical flavor

Starters are selected depending on:
 Ability to produce Flavour compounds, e.g. acetaldehyde, acetoin.
 Citric acid fermentation – which metabolize citric acid to produce aroma volatiles
such as diacetyl.
Examples of Flavour producing cultures
 Leuconostoc. mesenteroides subsp. cremoris
 Leuconostoc. mesenteroides subsp. dextranicum
 Leuconostoc. lactis

f) Culture compatibility- some strains are compatible, while others antagonistic

g) Product to be made and its properties

Different products have different end products . Many Lactic Acid Bacteria( LAB)
produce exopolysaccharides (EPS), which may provide viscosifying, stabilizing, and
water-binding effects in cheeses. EPS have also the potential to be used as surface
carriers of bacteriocins or bacteriocin producing LAB, and species such as Leuconostoc
mesenteroides, Streptococcus mutans and several lactobacilli (Lactobacillus brevis,

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 Lactococcus lactis subsp. lactis, L. lactis subsp. cremoris, Lactobacillus casei, Lb. sake,
Lb. rhamnosus,) and thermophilic (Lb. acidophilus, Lb. delbrueckii subsp. bulgaricus,
Lb.helveticus and S. thermophilus) are known to produce EPS.

h) Proteolytic and Lipolytic activities- ability to produce peptides, amino acids, fatty acids
etc- common in cheese making.
 Bring about coagulation of protein and form gel.
 The proteolytic activity of starter culture is important because it leads to;
 Liberation of peptides and amino acids, which affect the physical structure of the
product
 Many amino acids produced are essential for the growth of several cultures and
 Peptides and amino acids act as flavor precursors.

i) Production of other compounds- like CO2, alcohol, propionic acid, which are essential
in products like kefir, Swiss cheese
 Production of bacteriocins which prevents the growth of pathogens, as well as
many spoilage organisms.

j) Vitamins metabolism- some vitamins may be synthesized leading to increased

nutritional content in fermented milk. This increase or decrease depends greatly on the
strain of starter.

k) Possesses therapeutic significance

Such as:
 Improvement of intestinal organisms,
 Produce antibacterial substances, and
 Improve immunity

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Quality tests on starter cultures
A. Purity of Starter Cultures
Pure starter cultures - Absence of contaminants, pathogens, and extraneous matter.
The purity of the starters is evaluated by using microbiological, microscopic or chemical tests
i) Microbiological
Tests for the contaminants like coliforms, yeasts and molds (both should be absent in 1 ml of
culture). Starter must be free from foreign bacteria, yeasts and molds. Plate count and
selective plating methods are employed.
ii) Microscopic examination
This is performed by Grams staining or Newman’s stain. Gram -ve bacteria, spore formers,
yeasts and molds and staphylococci etc. should be absent. The lactic bacteria should appear
as Gram +ve cocci or thin rods.
iii) Catalase test
Add few drops of 3% hydrogen peroxide to starter culture. Presence of effervescence
indicates possible contamination of the given culture. Lactic acid bacteria are negative for
catalase and thus catalase positive test indicates contamination of starters

B. Activity of starter cultures

i) A good starter should have the ability to produce lactic acid at vigorous and steady rate.
i) It should be able to grow rapidly in suitable organic substances
ii) It can be easily cultivable in large quantities
iii) It should be able to maintain physiological constancy
iv) It should be able to produce necessary enzymes readily and profusely in order to bring about
the desired chemical changes
v) It should have the ability to carry out transformation under comparatively simple and
workable modifications of environmental conditions

Activity rating of starter cultures is determined through acid-producing and other functional
activities. Chemical and microbiological parameters are used for assessing the activity of
starters:

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i) Titratable acidity
A titration value of 0.4 %LA or higher indicates that the culture should be well suited for
acid production. The cultures lose their activity due to acid injury if they are allowed to over
ripen. A developed acidity in the range of 0.7 to 0.85% LA is optimal.
ii) Creatine test
The Creatine test makes it possible to quickly secure general information on the comparative
amounts of acetyl methyl carbinol plus diacetyl. It is a common method used for indicating
diacetyl content in flavor producing mesophilic cultures (buttermilk, cultured cream, cottage
cheese etc.).The cultures form a pink colored complex with Creatine.
iii) Viable cell count: a microbiological test to estimate the number of viable starter cultures
cells. A high number per ml is desirable
iv) Dye reduction test
An excellent culture reduces the resazurin dye or methylene blue dye in 35 min and a fairly
good culture takes 50 to 60 min.

C. Other tests for commercial starter cultures

Package integrity, accuracy of label information on the package and Shelf life of the product
according to specification are also verified before the starters are accepted for use

Some Commercial Dairy Starter Culture Producers throughout the World

Producer Country Website
Christian and Hansen Denmark http://www.chr-hansen.com
Danisco Denmark http://danisco.com
DSM The Netherlands http://www.dsm.com
Alce Italy http://www.mofi nalce.it
Centro Sperimenti del Latte Italy http://www.csl.it
Valio Finland http://www.valio.fi
BioSource Flavors, Inc. United States http://www.biosourcefl avors.com
CSK Food Enrichment The Netherlands http://www.cskfood.com
BIOPROX France http://www.bioprox.com

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Preservation of starter cultures
a. Storage and preservation of cultures

Starters are preserved in liquid, frozen and dried forms.

Liquid sub-cultures are for routine use, while frozen and freeze dried forms are for long term
preservation.

i) Liquid (lyophilized) starter: are nowadays quite rare as used to propagate mother
culture. Are kept refrigerated and are transported in insulated polystyrene boxes packed
with dry ice; time in transit should not exceed 12 hours.
ii) Dried starter: unconcentrated (spray dried or freeze dried/lyophilized; some of these
methods are rather old and not used at the present time), Concentrated freeze dried. freeze
dried DVS are preserved at -18oC for up to 24 months

iii) Frozen starter: Frozen at -20°C (un concentrated), Deep frozen at -40°C to -80°C
(concentrated), or Ultra-low temperature freezing at -196°C in liquid nitrogen
(concentrated).

Factors that affect starter culture viability

1) Temperature
Temperature directly affects the growth of microorganisms. The majority of lactic starters such
as L. lactis subsp lactis, L. lactis subsp cremoris, etc. grow optimally at 27-32oC.On the other
hand, S. thermophilus and some lactobacilli grow best at 37-42oC temperature range. However,
Leuconostocs have their optimal temperature at 20- 30oC. This variation in optimal temperature
requirement may be a very important factor in strain dominance in mixed and multiple starters.

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 2) pH
The pH influences the growth and metabolic activity of lactic starter cultures during milk
fermentations. Lactic acid bacteria produce lactic acid at the level of more than 10 per cent of
their weight per minute after their growth in milk and thus, pH of milk is lowered. The extreme
acidic pH could be detrimental for the viability of LAB. pH below 5.0, causes considerable
injury to the cells of the starter culture which could adversely affect the performance of these
organisms. Control of pH in milk during propagation of starters can be achieved by using low
lactose milk. In externally pH controlled whey medium, the pH drop is controlled by an
insoluble buffer so that the lactic acid produced is immediately neutralized. Another promising
and practical approach to tackle this problem could be through genetic manipulation of these
organisms by introducing a pH sensitive promoter for regulation of structural genes involved in
acid production.

3) Strain Compatibility
Mixed starters have been used for the preparation of several fermented dairy products. Repeated
subculture of mixed strains of Lactococci may result in decrease in number or loss of all but one
of the strains that eventually a single strain remains in the mixed starter preparation. Some of the
factors responsible for strain dominance or overgrowth in a mixed culture by one strain include:
 Differences in generation times
 Acid sensitivities
 Production of antibiotics or bacteriocins by the component strains
 Differences in optimum temperature and rates of plasmid loss

4) Growth Medium
The media used for the cultivation of LAB are quite complex. The most widely used growth
media are MRS, M-17 and Lactic or Elliker’s medium. Apart from these media, lactic acid
bacteria also grow very well in milk. However, it is pointed out that during specific seasons of
the year, more inoculum may be needed during milk fermentation in order to achieve the same
type of acid production obtained with lower inoculum rates at other times of the year.

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 5) Inhibitory Substances
The growth and activity of the starter cultures in milk is adversely affected due to the presence of
residual antibiotics and sanitizers in milk as well as the production of antibiotic-like substances
(bacteriocins) by certain wild strains of Lactococcus lactis subsp. lactis and other lactic cultures
in raw milk. Antibiotics such as penicillin or streptomycin may enter milk as a result of their
indiscriminate use in the treatment of mastitis or udder diseases. Hence, milk must be thoroughly
monitored for the presence of residual antibiotics before addition of starter cultures.

6) Bacteriophages

Bacteriophages are considered as one of the single most important factors causing slow acid
production by lactic acid bacteria in the commercial environment. The most promising solution
to this problem could be by replacing the phage sensitive strains with phage resistant one.

7) Incubation Period
The period of incubation can affect the growth of lactic acid bacteria. Normally, 16-24 hr
incubation is adequate for the maximal growth of organisms at their optimal temperatures
however; storage of the ripened starters for about 18 hrs at low temperature does not affect their
activity. Over-ripened cultures are adversely affected on prolonged storage.

8) Heat Treatment of Milk

Heat treatment of milk usually improves its value as a medium for starter organisms and other
lactic acid bacteria. Adequate heat treatment of milk drives out the dissolved oxygen, brings
about formation of sulphydryl compounds (acting as growth factors), destruction of inhibitory
substances naturally present in milk and killing of antagonistic bacteria. However, with more
severe heating, slight protein breakdown may occur with the formation of peptides and amino
acids, which act as nutrients. In addition to these, different species of lactic acid bacteria appears
to behave differently in heat-treated milks. For example, the growth of S. thermophilus appears
to be favored, while L. lactis subsp. Cremoris disfavored by drastically heat treatment of milk.

9) Degree of Aeration

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The lactic acid bacteria are indifferent to aeration of the medium or slightly prefer a reduced
oxygen tension. Acid production is faster at the bottom of the container or under reduced oxygen
tension condition. Although excessive aeration may be the cause of slow starters, its effects may
be neutralized by heating milk or by adding sulphydryl compounds

10) Effect of Carbon Dioxide

A minimum concentration of carbon dioxide is essential for the initiation of bacterial growth.
Complete removal of carbon dioxide from a medium results in extended lag phase until the
bacteria have slowly produced sufficient carbon dioxide to sustain normal growth. For most of
the lactic acid bacteria, the optimum initial concentration of CO2 varies from 0.2 – 2.3% by
volume. Sterilized skim milk may contain only 0.3-0.5% of CO2and this may account for the
prolonged lag phase of the given starter culture. However, incorporation of yeast extract in milk
at a concentration of 0.5% can get rid of this problem.

11) Storage conditions

Storage conditions of lactic acid bacteria affect their performance during the manufacture of
fermented milk products. Storage in presence of acid will result into cellular injury and promote
the loss of plasmids. Therefore, it is important that during the maintenance of such cultures, they
should be transferred to fresh milk and in refrigerator without incubation. However, mature
cultures may also be stored at 2-5oC either in milk with added calcium carbonate. Cultures can
also be frozen and stored at -40oC or below in the freeze-dried state. Frozen storage in liquid
nitrogen (-196oC) in the form of starter concentrates also affords stability to cultures.

Raw milk quality for fermented milk production

Raw milk should be free from the following: -
i) Physical dirt - Should be clean, if not sieve or clarify
ii) Antibiotics – antibiotics prevent growth of bacteria in the starter culture
iii) Colostrum - the high amount of whey proteins causes wheying off resulting in thin
watery product. They also act as natural inhibitors in milk.

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 iv) Mastitis - low sugar (lactose) in mastitis milk does not allow production of enough lactic
acid resulting in a flat tasting product
v) Water - added water prevents formation of a firm coagulum.
vi) Chemicals- e.g. detergents, sanitizers, preservatives, which are injurious to the consumer
or will prevent growth of bacterial culture.
vii) Microbial quality: - should be low in initial microbial load. Some microorganism’s
produce bacteriocins which might prevent fermentation or be in competition with the
starter culture

Raw milk quality testing

The quality of milk can be ascertained by carrying out milk tests. These tests assess the
following
 Freshness
 Milk abnormalities
 Hygienic quality
 Extraneous matter
 Physical characteristics
 Inclusion of “illicit” substances

Common milk tests

1) Organoleptic/sensory evaluation
This is done by use of common human senses mainly sight and smell.
Acceptable milk should be:
 Milk in a clean container
 Have a characteristic cowish smell
 Creamish white to yellowish white in color
 Clean in body and texture
 Have the right viscosity, Contains no clots , lumps or any foreign matter

Procedure for sensory evaluation of milk in cans:

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 i) Check cleanliness of the outside of the can
ii) Open the can, smell and check the cleanliness of the lid/milk
iii) Stir/shake milk for decanted materials and dissolved odor to come up.
iv) Take the smell again
v) Check the color and the consistency
vi) Touch the container sides to determine time of milking by temperature estimation.
vii) Make a decision on accepting or if suspect carry other objective tests.

2) Alcohol Test
It checks the stability of milk when mixed with equal volume of 68% alcohol for one minute.
This test is more sensitive than Clot-On-Boiling. It detects developed acidity of 0.23% Lactic
Acid. You can make this test more sensitive by: Increasing the concentration of alcohol, e.g.
80% or using double volume of 68% alcohol.

Procedure for Alcohol test:

i) Measure 1 or 2 ml of milk into a receptacle.
ii) Add equal or double volume of 68-70% ethyl alcohol.
iii) Mix thoroughly.
iv) Check whether the milk has precipitated.
v) Normal milk does not clot.
Alternatively use alcohol gun automatic measure of milk and alcohol

3) Lactometer test
It checks the wholesome of milk through determining its density. The normal density of milk is
1.028 to 1.032 at 20oC (KeBS std). Change of milk composition through adulteration (addition of
water/substances or removal of cream) changes the density of milk. Removal of cream increases
the density to beyond 1.032 while addition of water reduces it to below 1.026.

Procedure for Lactometer Test:

i) Put milk into a lactometer jar.
ii) Note its temperature (MT).

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 iii) Note the Lactometer Calibrated Temperature (LCT).
iv) Lower the lactometer into the milk sample gently and allow it to settle.
v) Take the lactometer reading (0L) corresponding to the meniscus of milk
vi) Calculate the Correct Lactometer Reading (CLR)
CLR = (MT – LCT) 0.2 + LR
vii) Calculate the specific gravity:
Specific Gravity = 1.0 + (CLR ÷ 1000)

4) Titratable Acidity Test

This test measures the amount of acid (sourness) in milk. Normal fresh milk has a natural acidity
of 0.14 – 0.18% lactic acid. Above this is due to development of acidity through microbial
activity.

Procedure for Titratable acidity

i) Put 9ml of milk in the beaker or petri dish.
ii) Add 3 – 5 drops of 5% phenolphthalein indicator.
iii) Titrate with 0.1N NaOH solution to a faint permanent pink colour.
iv) Read the volume of NaOH used.
% Lactic acid = Volume of NaOH used in mls / 10

Factors that cause inhibition of starter cultures

1. Residual antibiotics in milk as a result of mastitis treatment. Starter cultures are very
susceptible to residue antibiotics; Lactococcus genus are very sensitive while
Leuconostoc are less sensitive
2. Residual detergents and sanitizers- due to cleaning and sanitization of dairy equipment. It
can be due to human error or failure in the automatic cleaning cycle.
3. Bacteriophages- viral pathogens that destroy bacterial starter culture. Once attacked by
bacteriophages, it is very hard to control. Basing onn this, starter culture can be
classified into;

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(i) Phage insensitive- not attacked by any phages
(ii) Phage carrier-slight reduction in activity once attacked
(iii) Phage sensitive- complete destruction once attacked
Bacteriophages are host specific eg phages that attack L.L lactis cannot attack L.L cremoris.
Phage attack is easily recognised by low acid production or low aroma production. Whey is a
major source of phages especially in cheese rooms and therefore whey should be removed from
the cheese room through a closed system.

Control of phages
1. Heat treatment of bulk starter culture milk to > 900 C for 20 minutes . They are very heat
labile hence easily destroyed by high pasteurization temperatures.
2. Adhere to strict hygiene measures during starter culture propagation e.g. asceptic inoculation
technique, use of sterile equipment, filtration of air, avoid movement of personnel and
equipment to starter culture room from production room
3. Locate starter culture room away from the production room. A foot bath should be provided
before the starter culture room; hands should always be washed with a disinfectant before
handling any starter culture
4. Use of culture rotation system- rotate the starter cuture so that the culture that is not attacked
is used
5. Use of phage resistant or unrelated strains
6. Production of phage-free bulk starter cultures
7. Use of phage inhibitory media( PIM)

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 Processing of yoghurt
Definition:
Yoghurt is a semi-solid fermented milk product obtained by fermentation of heat-treated milk by
the action of symbiotic blend culture of Streptococcus Salivarius subsp. thermophilus (ST) and
Lactobacillus delbrueckii subsp. bulgaricus (LB) (Codex Alimentarius, 2003)

Introduction
Yogurt (also spelled yogourt or yoghurt) originated centuries ago and has evolved from many
traditional Eastern European (e.g., Turkish and Bulgarian) products. The word is from the
Turkish Yogen, meaning thick.
Yoghurt is the best known of all cultured-milk products, and the most popular almost all over the
world. Consumption of yoghurt is highest in countries around the Mediterranean, in Asia and in
Central Europe.

Classification of Yoghurt
Yogurt is mainly classified based on:-
 Chemical composition - they are classified as full-fat, reduced-fat or low-fat, fruit
yoghurt
 Method of production- they can be grouped as set, stirred, drinking
 Flavor type- can be classified into subgroups such as Plain/natural yogurt, flavoured
yoghurt (strawberry, vanilla etc)
 Post-incubation process - can be grouped as concentrated, dried, and frozen, long life
 Textural characteristics – can be grouped as thick yoghurt, thin ,smooth

Types of Yoghurt
i. Plain/ natural yoghurt: Plain or natural yoghurt has no added ingredients or additives other
than the starter cultures

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 ii. Flavoured yoghurt Yoghurt with various flavouring and aromatic additives
iii. Stirred yogurt: Yogurt is first made in a large container and then spooned or dispensed into
secondary serving containers. The consistency of the “set” is broken and the texture is less
firm than set yogurt. This is the most popular form of commercial yogurt.

iv. Set, firm or eating yogurt: the yoghurt packaging into cups is done immediately after
inoculation, then incubation and overnight cooling. The yoghurt is served while in the cups
and eaten with a spoon like ice cream. There is no stirring

v. Drinking sweet yogurt: Stirred yogurt to which additional milk and flavors are mixed in. Fruit
or fruit syrups are added to taste. Milk is added and mixed to achieve the desired thickness.
Has total solids content not exceeding 11%. The shelf life of this product is 4–10 days, since
the pH is raised by fresh milk addition. Some whey separation will occur and is natural.

vi. Fruit yogurt: The preparation method is similar to that of either drinking or set yoghurt. The
difference being that instead of flavours, real fruit, fruit pulp, fruit syrups, or pie filling are
added. The type of fruit should be sweet and pulpy. The fruits used include; bananas,
pineapples, papayas, mangoes, oranges, passion fruits, tree tomato fruits, strawberries etc.
They are placed on top, on bottom, or stirred into the yogurt

vii. Frozen yogurt: it can be manufactured in two ways. Either yoghurt is mixed with ice cream mix
or yoghurt is frozen by batch or continuous freezers. Frozen yoghurt can be divided into soft-
served and hard frozen types.

viii. Dried yogurt (Kurut in Turkey): Yogurt is sun dried for longer preservation.

ix. Concentrated yoghurt: incubated in tanks, concentrated and cooled before being packed. DM
of the product is increased after fermentation by drained off the whey from the coagulum.
This type yoghurt is sometimes called strained yoghurt, sometimes labneh, labaneh

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 x. Health Yoghurt
The preparation method is similar to that of either drinking or set yoghurt. The difference is the
type of being starter culture used. This yoghurt is made by use of cultures containing the
following bacteria:
 Lactococcus salvaricus ssp thermophilus for acid production
 Lactobacillus delbreuckii ssp bulgaricus for flavor (acetaldehyde, diacetyl) production as in
the above yoghurts plus one, two or the three healthy bacteria listed below: -
 Lactobacillus bifidus
 Lactobacillus acidophilus
 Lactobacillus casei ssp rhamnosus
 Bifidobacterium bifidus

xi. Diet Yoghurt

This is low calorie/energy yoghurt. The difference is that it is prepared from skim milk and no
sugar added. The starter culture used can either be for ordinary yoghurt or healthy yoghurt.

xii. Long-life yoghurt

This is sterilised yoghurt that can be transported or stored at room temperature. The shelf life of
the product is extended through HTST Heat treatment of the finished product, either immediately
before packing or in the package. Heat treatment of yoghurt prolongs its shelf life by:
 Inactivating the starter bacteria and their enzymes
 Inactivating contaminants such as yeasts and moulds
Heating to 70 – 75°C kills all the virulent micro-organisms in the yoghurt. In many countries
yoghurt is defined as a product in which the microbiological flora is kept alive right up to the
instant of consumption. This means that heat treatment of the end product is prohibited

xiii. Yogurt cheese: It is a fresh cheese made by draining overnight by separating the whey. The
flavor is similar to that of a sour cream with the texture of a soft cream cheese. A liter of
yogurt will yield approximately 500 mL of cheese. Yogurt cheese has a shelf life of
approximately 7–14 days when wrapped and placed in the refrigerator and kept at less than
4°C.

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Yoghurt Ingredients and Additives
Although milk of various animals has been used for yogurt production in various parts of the
world, most of the industrialized yogurt production uses cow's milk. Whole milk, partially
skimmed milk, skim milk or cream may be used. Other yogurt ingredients may include:

1. Other Dairy Products: concentrated skim milk, non-fat dry milk, whey, lactose. These
products are often used to increase the non-fat solids content
Caseinates e.g. whey protein concentrates and co-precipitates are usually added to increase the
protein content of the product. Addition of more than 2% of caseinate will result into
undesirable and uncontrollable thickening of the product. Addition of co-precipitates favourably
affect the consistency and viscosity of the product and will also prevent wheying off

2. Sugar or Sweetener
The disaccharide sucrose, or a monosaccharide such as glucose, can be added alone or in
conjunction with fruit addition. To satisfy dieters, among whom diabetics are an important
category, sweeteners should be used. A sweetener has no nutritive value but tastes very sweet
even in very small doses.
The fruit in question usually contains about 50% sugar or a corresponding amount of a
sweetener, so the required sweetness can normally be supplied by adding 12 to 18% fruit. It
should be noted that adding too much sugar (more than 10%) to the milk before the
inoculation/incubation period has an adverse effect on fermentation conditions because it
changes the osmotic pressure of the milk.
Addition of sugar should be followed by intensive stirring . The sugar can be added in granular
form of syrup( 60-5%), however, syrup will ead to dilution of the milk hence reduced total solid
content. Sugar should be added to the milk before pasteurization because;
(i) The heat treatment of milk will destroy most of the mesophilic moulds and yeasts in
sugar
(ii) The consistency of the product is better when sugar is added before pasteurization since
there is no excess damage to the coagulum during stirring

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Types of Carbohydrate Sweetener
i. Sucrose (saccharose) - sucrose has the empirical formula C12H22O11 and the refined
carbohydrate is obtained commercially from sugar cane or sugar beet. It is widely used in
the food industry as a sweetening agent and can be obtained in a granulated or syrup
form.

ii. Invert sugar – is a mixture of glucose and fructose obtained by splitting the
disaccharide sucrose with molecular formula C12H24O12

iii. Fructose – also called fruit sugar is simple sugar with the molecular formula C6H12O6

iv. Glucose – also called dextrose is simple sugar with the molecular formula C6H12O6

Any of these different types of sweetening agents could be employed for the manufacture of
yoghurts and the choice of any one particular sugar is determined by one or more of the
following factors:

a) Availability and cost of the sweetening compound- should be easily available and
affordable: for these reasons it is probable that sucrose is the most widely used.

b) Legal aspects- should be permitted as a food additive: whether a certain sugar is

permitted as a food additive, although since most sweetening agents are derived from
natural products, with the exception of the artificial sweeteners, prohibition is unlikely.

c) Storage facilities- should be storable in the available facilities: Syrups are mainly stored
in large metal containers or silos. Granulated products are stored in multilayer bags.

d) Nutritional aspects- type of consumers i.e. for “diabetic”, it should provide sweetness
and low calorie: fructose is a very sweet sugar and a sucrose/fructose syrup mixture used

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 at a low level can provide both sweetness and a reduced calorie intake; in addition,
fructose, like sorbitol, is absorbed only slowly into the bloodstream and its use in
“diabetic” yoghurt production is a clear possibility.

3. Flavors
A variety of different flavoring ingredients (fruits, natural flavors and/or synthetic flavors) are
currently added to yoghurt.

Fruits and fruit flavors (common examples)

 Apricot
 Black cherry
 Blackcurrant
 Peach
 Pineapple
 Raspberry
 Strawberry

Flavoring agents
 Natural flavors and flavoring substances (botanical origin)
 Nature-identical flavoring substances (botanical origin)
 Artificial/synthetic substances (chemical origin)

4. Colors
Color is added to fruit and flavored yoghurts to make the products more attractive. The active
agents may be naturally derived, nature identical, caramel or artificial.

Permitted food coloring matter arising exclusively from flavoring substances

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 5. Stabilisers
Hydrophilic colloids can bind water. They increase the viscosity and help to prevent whey
separation in yoghurt and to improve viscosity and consistency of the product. The type of
stabiliser and the rate at which it should be added must be determined experimentally by each
manufacturer. The product may acquire a rubbery, hard consistency if the wrong stabiliser, or an
excess of stabiliser, is used. Correctly produced, natural yoghurt requires no addition of
stabilisers, as a firm, fine gel with a high viscosity will occur naturally. Stabilisers can be used in
fruit yoghurts and must be used in pasteurised yoghurt. Stabilisers (0.1 – 0.5 %) such as gelatin,
pectin, starch and agar-agar are the most commonly used substances. Others include;
carboxymethyl cellulose( CMC), locust bean gum, alginates, carrageenans, whey protein
concentrate

Some ingredients that can be used in manufacture of yoghurt

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 6. Starter Culture
The starter culture for yogurt production is a symbiotic blend of Streptococcus salivarius subsp.
thermophilus (ST) and Lactobacillus delbrueckii subsp. bulgaricus (LB). Although they can
grow independently, the rate of acid production is much higher when used together than either of
the two organisms grown individually. ST grows faster and produces both acid and carbon
dioxide. The formate and carbon dioxide produced stimulates LB growth. On the other hand, the
proteolytic activity of LB produces stimulatory peptides and amino acids for use by ST. These
microorganisms are ultimately responsible for the formation of typical yogurt flavour and
texture. The yogurt mixture coagulates during fermentation due to the drop in pH. The
streptococci are responsible for the initial pH drop of the yogurt mix to approximately 5.0. The
lactobacilli are responsible for a further decrease to pH 4.0. The following fermentation products
contribute to flavour:
 Lactic Acid
 Acetaldehyde
 Acetic Acid
 Diacetyl
Average amounts of ingredients added in yoghurt making
These may be included at the following ratios:
i. Sugar 4.0 -6.0%
ii. Skim milk powder 1.0 – 3.0%
iii. Starch 1.0 – 2.0%
iv. Pectin/gelatin 0.03%
v. Food colour according to consumer preference

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