Dairy Microbiogy Notes PPT11

DAIRY MICROBIOLOGY
Microbiology of Cream
 Cream may be defined as "that portion of milk which is rich in
milk fat" or
 when milk fat is concentrated into a fraction of the original

milk that portion is known as cream”
 NS – Not less than 18% Fat

Classification of Cream
 Double Cream: Double cream has a very high butterfat content;
48%.
• It is very rich, heavy and used specifically for puddings and
desserts, and piping.
• The more double cream is whipped, the more thicker it gets. It
contains absolutely no thickening agents.
 Clotted Cream: Clotted cream is also popularly known as
Devonshire cream or Devon cream.
• Fat content >55%.
• This cream has a slightly caramelized flavor, as obtained by
heating un-pasteurized cow's milk, which is then left in shallow
pans for several hours.
• The cream content rises to the surface and gets clotted.
 Whipped or Whipping Cream: Fat content for both whipping as
well as whipped is around 35%.
• Whipped and whipping cream are made by mixing cream with air
to roughly double the volume.
• The air bubbles are captured in a network of fat droplets.
Half and Half Cream: It is usually used in tea and coffee.

• Half and half usually has 10 - 18% fat.
• Use of 'half and half' is quite popular in recipes that call for the
cream's flavor and texture without its richness or thickness.
• Half and half is referred as a equal mix of whole milk and full cream.
• This cream will not whip or thicken, and the consistency will be
more like full fat milk.
o Single Cream: Light cream or as coffee cream.
• Contains very little fat; between 18 - 30% and is very light in texture
• It will not whip, used to poured over puddings, in coffee or dishes that
call for very light cream flavor
o Sterilized (or Canned) Cream: Must contain not less than 23% fat.
• Sterilized half cream not less than 12%
o Sour Cream: Can be both thick and light, added with souring culture to
give sour taste and flavor.
• Culture is added to the cream which is heated to about 20° C for 10 - 14
hours.
• Heating produces lactic acid responsible for the sour taste.
• Light sour cream too is made in the same way, but has only 18% or less
milk fat.
• used in sauces and dressings, casseroles and cakes or served on
Cream Minimum fat content Use
Half cream Contains no less than 12% milk fat (Not sterilised) In coffee, pouring on fruit and desserts
In coffee, pouring on fruit and desserts,

Single cream Contains no less than 18% milk fat (Not sterilised)
adding to soups and savoury recipes
Pouring/spooning cream for desserts,

Double cream Contains no less than 48% milk fat can be whipped for piping onto cakes
and pastries
Aeration for applications including

Whipping cream Contains no less than 35% milk fat
desserts, cakes and pastry fillings
Contains no less than 35% milk fat (Cream has been

Whipped cream For dessets, cakes and pastry fillings
whipped)
Used in the classic English cream tea

Clotted cream Contains no less than 55% milk fat (Cream is clotted) and as a dessert cream. Virtually unique
to Cornwall, Devon and Somerset
Sterilized cream Contains no less than 23% milk fat (Cream is sterilised) On or in desserts
Cream Preparation
Production of Transport to Storage in

milk Separation
the dairy the dairy
Heat Standardizati
Packaging Storage on
treatment
Storage and
Sale
distribution
Production on the farm:
 Hygienic quality of milk is important because although
vegetative cells are killed by heat treatment but spores are
not.
 If spore content is high (>100 ml-1) it may be worthwhile to

reduce this by high-speed centrifugal method, as aerobic
spore formers tend to form chains, these are more easily
removed than single cells.
Transportation to the dairy &
storage in the dairy:
 Milk should never be allowed to rise above 5°C in order to keep
the growth of psychrotrophs to a minimum, as they are very
important in long shelf life product.
 Cold stored milk does develop taints because psychrotrophs are
active against fats and proteins, but does not sour because
lactose is not attacked.
 Enzymes survive pasteurization and cause changes; so
prolonged storage (>48h) is not advisable.
Separation:
 Mechanical centrifugal separator, at a temperature between 40
to 50°C ideal for growth of bacteria.
 Higher temperature of above 50°C can be used but the fat losses
will be more.
 Advantageous to use modern equipment separating milk at low
temperatures e.g. 25-30 °C
• Additional advantage of causing less physical damage to the fat
globules.
 Separation also removes visible dirt, somatic cells and other
foreign matter in the form of slime.
 Some bacteria, especially clumps and spores are also removed.
 Many bacteria are removed as evidenced by high counts in
slime.
Standardization:
 Cream is standardized by
• Separating the milk at slightly higher fat content than

required and
• Standardizing by adding calculated quantity of skim milk.
 Cream should not be held for long and quality of skim milk
should be good.
Homogenization:
 Homogenization necessitates
• An extra treatment and so extra contamination
• May break bacterial clumps.
• Requires holding the cream at higher temperature(warm) for
a period.
 Homogenization should take place immediately before the
final stage of heat treatment.
 So final heating of cream “Increased surface area increases
action of lipase”
Heat treatment:
 Pasteurization: LTLT 65°C / 30 min or HTST 74°C / 15 sec, 82°C
/ No hold UHT 140°C / 2 sec
The heat treatment applied for cream should meet the following
criteria
 Destruction of all pathogens

 Achievement of desired shelf life
 Avoidance of cooked taints, which result from the production
of volatile sulphur compounds when milk and cream are
heated above 80°C.
These usually disappear in 1 or 2 days.
 In destruction of milk enzymes, particularly lipases
Heat treatment:
 The pasteurization temperature of cream are relatively high
because of
 The protective effect of fat on the bacteria

 The slower heat transfer in cream
 The requirement of a long shelf life
 Tyndallisation i.e., heating at 100°C for 30 min is also

effective in destroying the spore formers if carried for 3
successive days.
Packaging :
 The cream should be packed in clean containers such as
cartons, jars or bottles
 Cartons whether of paper or plastics, are of good hygienic

quality due to their method of manufacture, provided stored
in a clean atmosphere
 Bulk quantities are transported in a stainless steel tanker
 Intermediate quantities are distributed in steel or aluminum

cans
Sterilized Cream / Canned
Cream
 After standardization, the cream is heated to homogenizing
temperature usually above 65 ◦C
 After homogenization at about 2500 lb/in2 the cream is filled
into cans at a temperature of 30-35 ◦C
 Cans are then heated at 116-121 ◦C for 30 mins
 Microbiological problem:
 B.subtilis spores being very heat resistant, these can germinate
and produce a bitter taint and thinning of cream by the
production of lipolytic and proteolytic enzymes.
 This fault is usually associated with
 Poor quality of milk high in spores
 Dirty equipment
 Non-spore former presence indicates contamination after
sterilization and in canned product, a defective can or leaker.
Sterilized Cream / Canned Cream
 Common water borne organisms

 Proteus associated with thinning and bitterness
 Coliforms associated with gas
 Cocci associated with acid curdling
 A few cans from each batch should be subjected to accelerated
storage tests by incubation at 37 and 55°C for at least 7 days.
 Spoilage can be detected by
 external examination i.e. swelling of cans or shaking them and
by internal examination for others
Frozen Cream:
 This is least objectionable method.

 Cream is pasteurized at 75 – 88°C / 15 sec and cooled to 1°C.
 Frozen quickly and stored at –18 to –26°C.
 Keeping quality of 2-6 months with an average 6 months is
possible.
 It does not reduce the number of bacteria.
 Few organisms may be killed by the formation of ice crystals.
Hygienic Control In Cream

Processing
Should extend from production of milk on the farm to the delivery to the
customer.
 Hygiene means prevention of contamination of the cream at all stages
 Shelf life of cream is controlled mostly by
 Cleanliness or sterility of equipment
 Temperature of the cream
 Air sterility near fillers – which is often neglected but most important.
• Dairy surrounded by vegetation (fruit trees) air may be rich in yeast and molds.
• Check test should be made near lines handling cream.
• The plates (malt agar or potato dextrose) are exposed for 60 min, in lines
handling cream.
• Total count not more than 60 and Yeast or Mold not more than 10
• Windows should be closed, rubber air lock doors fitted, filtering systems should
be installed.
 Water should be chlorinated at 5-ppm available chlorine and regularly checked.
Inline testing of Cream
Equipment
 Samples are taken at various points in processing line and
examine them by a suitable microbiological method. E.g.
 Exit from Pasteurizer

 Entry to storage tanks
 Exit from storage tanks
 Entry to filler
 Entry from filler
 Filled cartons
Keeping Quality / Shelf Life
 Keeping quality is of 14 days with the cream held at

temperature below 5°C.
 Causes for poor keeping quality:
1. Poor quality raw milk, particularly with high spore
and thermoduric counts.
2. Poor hygiene in separation
3. Wrong choice of heat treatment temperature
4. Poor hygiene in processing
5. Poor hygiene in packaging
6. Too high temperature of storage and distribution
Quality of the Raw Milk
 From the point of keeping quality in cream bacteria

found in Milk may be placed in three groups
 Thermo liable: Killed by pasteurization (71°C with 15s)
e.g. Coliforms, Pseudomonas
 Thermoduric : Do not form spores but survive ordinary
pasteurization e.g. Micrococcus, Streptococcus,
Lactobacillus, Bacillus, and occasionally gram-negative
rods
 Aerobic spore forming bacteria e.g. Bacillus cereus
Diagnosis of reasons for
poor
Results
quality raw
Interpretation
cream:
High count and high Inadequate heat treatment and or
Coliforms Unhygienic manufacture and or storage at
high temperature.
High count but low Coliforms Good hygiene but storage at high
temperature
Low count but high Coliforms Poor hygiene in manufacture but storage at low
temperature less then 5°C
Low count and low Coliforms Good hygiene except aerial contamination
but high Molds in dairy
Low count and low Coliforms Good hygiene except contamination from
but high yeasts fruits, directly or indirectly
Low count and low coliforms Cream made from milk having a high spore count
but high aerobic spores
Effect of temperature
 Types of bacteria according to temperature
 Psychrophiles (0-27 °C)
 Psychrotrophs (0-45 °C)
 Mesophiles (10-45 °C)
 Thermophiles (45-63 °C)
 Cream is refrigerated perishable food hence problem is to
minimize the growth of Psychrophiles and Psychrotrophs
 Two key temperature for their growth
-Below 6 °C growth is very slow and above 13 °C growth is rapid
- Low temperature bacteria ; 2 ranges of importance's from 0-4 °C
and 5-10 °C
Microorganisms causing defects
in cream
 Contamination Source
 Udder
 Cow
 Milking
 Original flora or Cow derived pathogens: Lactic and other
Streptococci, Micrococci, Corynebacteria and aerobic and
anaerobic spore forming bacteria
 Other organisms accumulate until final heat treatment
 Gram-ve rods from watery environment e.g. Psychotropic,
Staphylococci and Lactobacillus prominent depends on hygiene
and temperature of cream
 Bacillus cereus are favored by failure to cool the cream rapidly to
Microorganisms causing

defects in cream
Fresh cream predominating mos at 5 °C are
 Pseudomonas
 Alcaligenes
 Acinetobacter
 Aeromonas
 Achromobacter
 At 30 °C
 Corynebacterium
 Bacillus
 Micrococcus
 Lactobacillus
Microorganisms causing
defects in cream
 After holding for 5 days at 5 °C
 Non-fluorescent Pseudomonas predominant
 Corynebacterium and Micrococcus reduced
 Micro flora of cream depends on the types of post-
pasteurization contamination and length of time of
storage at 5 °C.
 Mean shelf life: 6.5-23 days
 At the end of shelf life Pseudomonas spp. predominates
TAINTS IN CREAM
 Taint is defined as a contaminant that is detectable by the human
senses and is unpleasant
 Two types of taints
 Non biological or Chemical – last on standing
 Biological or Microbial – intensify on storage
 Milk fat in cream is in emulsified form and has enormous surface
area and readily absorbs odors from atmosphere. So not to be
stored in a place where disinfectants, paints, varnish, scents are
stored
 Chemical taints – Oxidized taint – Occur in cream of even very
good microbial quality held at low temperature. Sunlight and
trace concentrations of copper can promote oxidation. Higher the
bacterial count the slower the development of oxidized taint
because bacteria consume oxygen and lower the OR potential of
TAINTS IN CREAM
Cheesy
Putrid
Stale
unclean
Old Cream
Rancid Flavors
Bitter
Sour
Yeasty
Origin of taints
 Abnormal milks – mastitis, late lactation, methods of feeding, weeds in
pastures.
 Failure to cool milk immediately after milking – which permits lipase and
other enzymes to act.
 High counts in milk and / cream due to unhygienic production
 Dirty cream separators and equipment failure to cool cream
 Failure to cool cream
 High temperature of holding during distribution and sale
 Use of stale milk or if Cream stale when sold.
 If taints are present immediately after separation they may be due to the
enzyme action on milk or Inherent to milk it self.
 If developed after pasteurization of cream, may be due to Dirty equipment;
Thermoduric bacteria; High holding temperature
Testing of Milk
 Tests are conducted at the point of sale for raw milk, raw
cream, pasteurized cream
1. Thermoduric bacteria
2. Lipolytic bacteria – tributyrin agar 5 days at 22°C
3. Caseinolytic -- caseinate agar 5 days at 22°C
4. Coliforms counts – high counts suggest dirty equipment
Specific organisms and
taints they produce
 Bitterness
 Number of organisms are responsible, due to
biochemical attack on proteins to produce peptones and
polypeptides.
 E.g. Proteus, gram negative rods; Some yeasts and
molds some associated growth is necessary like
Streptococcus lactis produces acid and then Rhodotorula
mucilaginosa produces bitter flavor.
taints they produce
 Pseudomonas
 Gm –ve, nonspore forming rods, attack proteins and fats
strongly but no effect on sugars
 Ps. fluorescence - Greenish pigment and rancidity
 Ps. fragi - Apple like ester taint
 Ps. putrefaciens – Putrid odor
 Ps. nigrifaciens - Black discoloration
 Ps. aeruginosa - Opportunistic pathogen grow at 42°C ,
resistant to antibiotics and quaternary ammonium
compounds
taints they produce
 Yeast: -
 Yeasts are not very common because they are non-lactose
fermenters.
 Few lactose fermenters ferment lactose, or sugar added in
whipped cream and cause yeasty and fruity flavor and obvious
gas resulting in foamy appearance.
 Yeasts are acid tolerant and grow even after retardation of
bacterial growth
 E.g.Torula cremorisTorula sphaerica
Candida pseudotropicalis
taints they produce
 Aerobic spores: -
 Important in sterilized cream clotted cream.
 Sterilization / UHT process kills all vegetative cells and clears
way for spore germination and heating gives a shock to the
spores which stimulates germination.
 B. cereus – survive pasteurization; grow at low temperature,
sweet curdling, bitty cream and proteolysis.
 The other organisms are B. licheniformis, B. coagulans, B.
subtilis.
 All produce bitterness and thinning in sterilized cream
Food poisoning: -
 Cream is usually more severely heat-treated and so
less chances of food poisoning.
 After heat treatment contamination is very serious but
not a problem if cream held < 5°C
Water contamination:
 Important organisms are Pseudomonas,
 If water is contaminated with sewage Salmonella –
source and other fecal types may be present
Microbiological tests
Test Property Measured
Plate or colony count Cells or clumps forming colonies under the

conditions of the test (medium,
temperature etc.)
Direct Microscopic count Cells (or clumps) taking the stain, including
living and dead cells
Dye Reduction tests Metabolic activity (reducing power) of
-Methylene blue active cells
-Resazurin
Increase in acidity or other Number of bacteria able to produce the
Metabolic products substance measured e.g. lactic acid
Modern refined instrumental Production of metabolites or changes in

methods for measuring physical properties
metabolites, increasing
conductivity, producing heat etc.
 Special methods: -
 SPC
 Yeast & Mold count
 Proteolytic and Lipolytic activity
 Farm separated cream is tested for Molds, WIA & BA
 Mold tests: -
 Microscopic method -- Mold mycelia count
 Macroscopic method -- Methylene blue borax test
(MBB)
 Visual mold test -- Modified MBB test
 Coliforms: -
 Significance – If present in raw cream they indicate
unsanitary conditions of production and handling.
 In pasteurized cream it serves as an index of post
processing contamination
Microbiology of Butter
 Butter may be defined as fat concentrate obtained by

churning cream, gathering fat into a compact mass and then
working it.
 Composition: NS
 Total Milk Fat ≥ 80%
 SNF ≤ 2%
 Water content ≤ 16 %
• However if mentioned in label water content could be up to 18
%
• It may contain common salt or Annatto or both or anti-
oxidants
• It should not contain any preservatives
Types of Butter
 Pasteurized cream butter
 Ripened cream butter or Cultured butter (Danish Style): Made
from use of starter bacteria added to the cream producing
lactic acid and flavors compounds like diacetyl
 Unripened cream butter
 Salted butter: Salted butter is the most common style of
butter found in supermarkets. It has, at most, 2% salt added
after the buttermilk has been drained off.
 Unsalted butter: Unsalted butter contains no added salt.
Reduced and low salt butters have about half the salt you'd
expect in regular salted butter.
Types of Butter
 Sweet cream butter (< 0.2 % LA) : Dutch Method or NIZO
technique:
• Flavors are added to sweet cream butter after manufacture
• Cream is not ripened but churned in usual way and sweet
butter milk drawn off
• A mixture of starters and culture concentrate is worked into
the butter at correct rate to yield butter with characteristics
of normal ripened cream butter
• Advantage; production of sweet butter milk, lower copper
levels in butter due to low non-migration of copper from
serum to the fat
Types of Butter
 Sour cream butter

 Fresh butter
 Cold storage butter
 Clarified butter/ghee
• Clarified butter/ghee is almost pure milk fat (at least
99.7%) and used mainly in cooking. This is because it will
reach much higher temperatures before it begins to
smoke or brown and there's almost no moisture to cause
spattering
Factors favoring or discouraging
the growth of Mos in Butter
 Cream micro flora impairing quality of butter

 Butter cultures to enhance flavor and keeping quality of
butter
 Contaminating micro flora as spoilage agents
 Disease causing pathogens
Butter is less satisfactory for
growth of microbes:
 LESS LACTOSE: growth of microbes is minimized because lack of
lactose the principal component of microbe attack
 LESS MOISTURE: reduced water activity detrimental for microbes
 In salted butter, SALT INHIBITS bacterial growth acting as
preservative
 Water is in DISCONTINUOUS PHASE and so it is present in the
form of fine droplets. Most of droplets of water are sterile because
number of microbes is far less in number in comparison with
number of droplets. Even if some of the droplets are contaminated
with microbes they can not travel through the fat phase,
minimizing the microbial spoilage
Manufacturing of Butter
Receiving the Milk Receiving the Cream
Neutralization
Pre-heating (35-40 °C) Grading
Weighing
Sampling
Testing
Separation Cream
Standardization
35% Buffaloes
40% for Cows
Pasteurization
82-88 °C/ No hold
Manufacturing of Butter
Pasteurization
82-88 °C/ No hold
or Vacreation
Cooling 20-22°C Cooling 5-10°C
Ripening 20-22°C
Ageing (5-10°C)
Churning (temperature. 9-11°C)
Washing
(2-2.5% of fat) salting and working

Ripening
 Butter cultures contain lactic acid producers such as L.

lactis ssp lactis and / or
 L.lactis ssp cremoris and aroma producers such as L.lactis
ssp lactis biovar diacetylactis. Leuc.citrovorum and Leu.
dextranicum are added.
Procurement of milk and cream
 The quality of butter is influenced by the nature and
quality of original milk.
 Previously butter was prepared often as a means of
disposable of milk of doubtful quality.
 Now quality awareness and demand has increased the
use of high quality raw milk for butter making.
 Milk is cooled to below 5°C and hold at that
temperature before separation.
 The milk temperature will critically affect the
efficiency of separation, the ideal temperature being
46-49°C
Suggested Standards (cfu/ml) for
cream for Butter Making
(Pasteurized)
Procurement of milk and cream
 The cream is pasteurized between 88 and 93°C. High heat
treatment may tend to increase nutty flavor due to formation of
sulfhydryl groups.
 Vacreation after pasteurization is used to get rid of undesirable
feed taints.
 The cream is cooled immediately after pasteurization. This is a
vital in the manufacture of quality butter.
 Rate of cooling has certain effects but it is the final temperature
and holding time which have effect on subsequent stages of
manufacturing.
 Cream is cooled and held between 3-5°C and aged for about not
less than 4h to allow extensive fat crystal network.
Ripened cream Butter
 For obtaining the ripened cream butter cream after pasteurization is
cooled to 16°C-21°C
 Inoculated with 4% mixed starter culture containing
 Acid- producers L. lactis ssp lactis or
L. lactis ssp cremoris
 Flavor producers Leuconostoc cremoris or Leuconostoc dextranicum
and
L. lactis ssp.lactis biovar diacetylactis
 Temperature for ripening: Summer- 16-18°C
Winter- 19-21°C
 Ripening carried out on 2-3 stages to facilitate cooling of highly
viscous ripened cream: Alnarp Process- assists cooling of cream and
modify hardness characteristics of final butter
Firm butter Soft butter
-Cool cream to 19°C -Cool cream to 6-8°C

-Inoculate culture hold at 19°C to -Hold at 6-8°C for 2hr to form intensive
pH of 5.2 crystal network in fat
-Cool to 14-16°C and hold for 2h -Warm to 19°C-25 °C and hold until pH
-Cool to churn temperature falls to 4.9
-Cool to 15-16°C
-Cool to churning temperature
Neutralization
 Neutralization changes the pH of the cream to levels of

near neutrality at which the heat used in pasteurization is
less lethal than it is for sour cream
 Poor quality of water used as solvent constitute bacteria
Pasteurization
 Heat treatment used is high >71.1°C/30min and Usually

high percentage i.e., 99.9% bacteria are destroyed in
cream.
 This is due to high temperature heat treatment and
because of predominance of acid producers which are
sensitive to heat
Churning
 During churning a number of factors exert influences on the
quantitative changes in the micro flora of cream, buttermilk and
butter.
 The agitation of cream during churning results in the breaking up
of bacterial clumps. Thus affecting the bacterial count not content,
unless organisms are contributed by the equipment.
 Most of the bacteria in the cream are retained in buttermilk
fraction; consequently the bacterial count of buttermilk is higher
than that of cream or butter
 In properly pasteurized cream viable yeasts and molds are rarely
found.
 Their presence in butter is there fore must be attributed to
contamination from equipment particularly the churn.
 Most of the mold mycelia are retained in butter and only tiny
fragments pass into buttermilk.
 Possibility of contamination through the equipment is more
Churning (Equipment)
in case of butter and this increase extensively with the
carelessness of cleaning of pipelines, walls, and valves.
 Butter churns in butter industry are important source of
organisms especially yeasts and molds.
 Wooden churns are highly contaminated and very difficult to
clean and sanitize.
 It is important that a churn be treated daily to prevent
microbial growth especially growth and
 Sporulation of molds that might be embedded in the wood.
Churning (Equipment)
 Molds are deeply penetrated into porous wood and into the
crevices and openings that form when a churn is allowed to
remain idle and to dry out.
 Wood is very difficult to sterilize at best.
 Microorganisms on the inner surfaces of the churn can be
destroyed with certainty only through extensive heat
treatments.
 It is important that a churn be treated daily is to prevent
microbial growth especially growth and Sporulation of molds
that might be embedded in the wood.
Cleaning churn
 Rinse the churn by revolving for 10mins when 1/3 fully water
at 48.3°C
 Drain
 Wash with water with washing compound with ½ full
capacity at 60°C and rotate for 15-20min
 Drain.
 Rinse with water at 87.8°C half full and rotate for 15min
 Drain for 5min
 Just prior to use, rinse with solution containing 200ppm
available chlorine.
Working
 Working of butter causes little quantitative change in micro

flora.
 The physical structure of butter that is created or
established by working process greatly influences the
microenvironment. So working has effect upon the growth
of bacteria in butter, rather than in the effect on numbers.
 The total volume of droplets and the percentage of droplets
that are free of bacteria are dependent largely upon the
number of bacteria in pasteurized cream.
 The contaminants added during processing and the degree
of water dispersion in butter. Number of droplets in butter is
very large, 10 to 18 billions/gram.
Working
 A large percentage of droplets must be sterile, even in high-
contaminated butter, because the number of bacteria would be
far less than number of water droplets.
 The availability of nutrients for growth of bacteria is limited in
butter having its moisture finely dispersed growth of bacteria is
limited to certain areas where droplets are large or where limiting
factors such as pH, salt concentration are not inhibitory.
 Organisms are more active in under worked butter. Occasionally
the moisture content may be too high and may necessitate
reworking to eliminate some of the water.
 Reworking tends to alter the physical structure of butter and thus
may affect moisture dispersion and growth of organisms.
 Relatively large droplets are formed in the mass of butter.
Working/Reworking
 If little growth has occurred in butter and there is no tendency to
aggregate moisture droplets during reworking, there should be a
finer dispersion of moisture and decreased food supply for the
organisms, the effect will be same as that of working.
 If moisture droplets unite during reworking the bacterial cells in the
larger droplets should have more food available due to
temperature relationship
 Considerable bacterial growth and lack of tendency of droplets to
collect during reworking, then organisms are distributed in more
droplets so that bacterial activity could be increased.
 Considerable growth and tendency for aggregation of moisture
into large droplets extensive growth should occur owing to
distribution of increased number of bacteria in areas favorable for
growth.

Salting
Uniform distribution of salt may result in many of the water
droplets having a salt concentration that is inhibitory to many
microorganisms salted butter will show marked decrease in total
bacterial, yeast and mold counts during storage.
 The presence of salt in butter tends to retard the development of
defects caused by specific organisms. This effect increases with
increased salt content.
 Water of butter originates principally from two sources
 Buttermilk included in the butter granules
 Wash water remaining in butter.
 Moisture droplets originating from buttermilk remains free of salt.
While those originating from wash water contain most of the salt.
 Poorly salt distributed butter, the organisms originating from
buttermilk causes defects.
Storage temperature
 Many microorganisms causing defects in butter tolerate relatively
low temperature and can grow at temperature slightly above the
freezing point;
 The rate of growth increases markedly with increasing
temperature.
 Little growth of microorganisms occur in butter held below 0°C
and
 None would be expected at –17.8°C, the temperature commonly
above 0°C growth conditions become increasingly favorable.
Keeping Quality of Butter
Temperature of Very good Keeping Good keeping Poor keeping quality

Storage (°C) quality quality
20 3 weeks 10 days 3 days
15 5 weeks 20 days 3 days
10 2 months 4 weeks 1 week
0 3 months 6 weeks 1-4 weeks
-12 9 months 6 months 1-3 months
-25 12 months 9 months 3-6 months

Miscellaneous factors
 Water used for washing and standardizing butter may contain
undesirable microbes.
 Chlorination up to 200ppm is recommended.
 Mishandled butter colors may act as a source
 Packing process and un-cleaned surface of equipment's such as ladles,
boxes, printing machine, parchment liners and wrappers.
 Mold can be prevented by the storage of butter in an atmosphere of
CO2 and in sealed containers, similar results occur with Nitrogen gas.
 Wash water is associated with surface taints.
 Wash water is the source of organisms like Ps. putrefaciens, Ps.
Fluorescens, Ps. mephitica, Ps. fragi and Coliforms.
 Air in the butter and packaging rooms normally caries mold spores
originating from the outside air or from its passage over contaminated
surfaces.
Bacteriological quality of
water for dairy purposes
Total colony count /ml
22°C 37°C
Satisfactory <100 <10
Doubtful 100-1000 10-100
Unsatisfactory >1000 >100

BUTTER CULTURES
 Butter has advantage over other fat rich products or spreads is
its flavor, which is accomplished by the cultures known as
butter cultures.
 Now days they are used as pure cultures.
 Added at the rate of 4% and incubated at 16-18°C
 in summer and 19-21°C in winter season
 Cultures: - L. lactis ssp latis biovar diacetylactis ,
L. lactis ssp lactis
 Others such as Leuconostoc dextranicum, Leuconostoc
citrovorum
 Addition of butter cultures to cream materially increases the
bacterial count, as many of these organisms are carried over
into the butter.
Methods of using Butter
Culture
1. Culture is added to the cream and the cream is held for
several hours or overnight and then churned
2.Culture is added to the cream at the time of churning
3.Culture is added at the time the butter granules are formed and
when worked into the butter.
 Flavor imparted to butter depends on the amount of flavor

formed during ripening period of cream or
 On the amount of flavoring compounds present in culture at
the time of its addition to the cream or to the butter granules.
Advantage of using butter
cultures:
1. Flavor production
2. Common off flavors of butter such as feed, oxidized,
neutralizer etc., may be masked by flavor development
3. Lowering of pH by cultures prevent growth of many
proteolytic and lipolytic bacteria and thus prevents the bacterial
defects such as cheesy, putrid, surface taints.
E.g. Ps.putrefaciens effectively inhibited by the addition of 5 or

10% of starter prior to churning.
Ps.fluorescens (effectively), Ps.fragi (slightly) retarded by
ripening to churning acidities not less than 0.35% L.A
Advantage of using butter
cultures:
 The inhibition is due to many metabolic products
including lactic acid.
 High serum acidity cannot be relied upon to prevent all
types of microorganisms.
 Acid tolerant bacteria yeast and molds show growth and
may be responsible for spoilage of butter.
 Salting and thorough working of butter in combination
with inhibiting effect of acid are to be depended upon to
increase the storage life of butter.
Disadvantages of using butter
culture
 Extreme care must be used in the propagation of culture
from day to day
 Proper equipment must be made available
 Prescribed routine for handling and transfer of cultures
must be followed
 Cost of labor, equipment may be more
 Price difference between butter made with or without
cultures is not encouraging
 In salted butter chemical deterioration may be accelerated
by the presence of acid oily and fishy flavors may develop
Survival of butter cultures
 Normally there is a gradual decrease in the butter culture
organisms in salted butter during storage.
 This occurs when salt and moisture are well dispersed
throughout the butter. There may be many areas where
environment is favorable for culture organisms.
 In unsalted butter favorable temperature may permit the

growth of butter cultures.
 This is accompanied by increase in serum acidity and
decrease in pH that indicates the growth of butter culture
organisms.
Changes in flavor compounds
during the manufacture and
storage
1.Churning
 Diacetyl content increases because of agitation, which favor
the oxidation processes leading to the formation of diacetyl.
 Subsequently, it decreases because a high percentage of
diacetyl and acetyl methyl carbinol is lost in the buttermilk
and wash water.
 In salted and unsalted butter, diacetyl and AMC are found in
greater concentration in the serum than in the fat.
 However a smaller percentage of total AMC than of the
diacetyl is contained in the fat.
Starter distillation
 Volatile compounds present in the butter culture may be
collected by distillation of cultures that are prepared under
favorable conditions to maximum production of flavor
compounds.
 These are available commercially. These are added directly to
butter granules at the time of working
 Advantages: -
1. Flavor intensity can be varied
2. A high flavored butter may be produced without affecting
keeping quality
3. More economical than butter cultures
4. Time saving
5. Problems such as starter failure and poor methods of handling
can be solved
 Disadvantages:
1. Lack of delicate and characteristic flavor. Have somewhat

coarse or sharp aromatic flavor
2. Keeping quality of unsalted and slightly salted butter due
to butter cultures is lost
BUTTER DEFECTS
 The normal flavor of the butter should be fresh, clean flavor,

known as ‘nutty’ flavor
 Flat or insipid flavor: - occurs in fresh butter and is due to

• Excessive washing of butter grains during manufacture
• Dilution of cream with water
• The initial stages of bacterial deterioration
 Medicinal Flavor: - is due to the result of

• Medicaments used in treating the cow
• Added chlorine compounds to milk or cream
BUTTER DEFECTS
 Putrefactive Taint: - This is the most common bacterial

defect and is caused by
• Pseudomonas sp. especially by Ps. putrefaciens
 Un-chlorinated wash water is major source followed by poor
sanitation methods
 It is caused due to decomposition of proteins
 It is also known as surface taint because initially involves
surface layers
 Rapidly develops in butter at a temperature of 4.4 to 7.2°C
 Compound responsible for the defect is isovaleric acid
 Control of Putrefactive taints: -
 Proper pasteurization of cream
 A satisfactory supply or water is very essential
organism is very sensitive for low concentration of
chlorine. So 5-10 ppm available chlorine is excellent
 Organism is salt sensitive 2% salt well distributed in
butter is quite effective
 Diacetyl in butter has effect in retarding the growth of
Ps.putrefaciens. But Mechanism is not clear
BUTTER DEFECTS
 Rancidity: -
 Oxidative, and Hydrolytic rancidity due to break down of fat.
 Oxidative rancidity is due to oxidation of double bonds in UFA
catalyzed by metals especially copper in presence of light.
 Hydrolytic rancidity is due to lipases produced by growth of post
pasteurization contaminations.
 Lipase hydrolyses fat to fatty acids.
 Short chain fatty acids –C4, C5, C8 contribute to the “off flavor”
(Butyric acid is most important). Even mild heat treatment
destroys lipase originated from milk.
 Important one is the prevention of activation of milk lipase prior to
pasteurization.
 Lipolytic organisms in butter: Pseudomonas sp; Ps .fluorescens, Ps.
Fragi, Alcaligenes andYeast and molds
 Control of Rancidity - depends on
 Low bacterial counts in the raw materials

 Minimal contamination
 Proper dispersion of moisture and salt in butter
 Low-temperature of storage
BUTTER DEFECTS
 Fruity Flavor: -
 It is due to early phases of activity of lipoytic organisms
and the most common organisms are Ps. fragi, Ps.
fluorescens .
 It results in ester like odour resembling apple, later
developing into intense fruity flavor.
 Organism’s origin is water, feed and soil and so gain
through utensils and equipment
BUTTER DEFECTS
 Malty Flavor: - is due to malty variants of

 L. lactis ssp. lactis and is due to faulty sanitation of
equipment
 Skunk Like Odour: - Causative organism is Ps. Mephitica
 Black Discoloration: -
 The Ps. nigrifaciens causes this defect due to discoloration of
butter by black pigment
 Optimum condition: 4°C, 1.5-2.5 % salt,
 Adverse: >25°C of salt or 5% NaCl in media
 Sources: - floors, drains, butter printing and wrapping tables
Plant Layout
Principle of Zoning
Milk Powder Factory
Zoning : Shoe-Change
Examples of two wet zones (1)
Example of two wet zones (2)
Make things easy
Unmanaged waste
Ventilation system
Open Process
Visitors Gallery
Visitors
Pest control
Pest control
Pest Control ?
Good Manufacturing Practice
Microbiology of Indigenous
Milk Products
Microbiology of Indigenous
Milk Products
 “Indigenous milk products” refer exclusively to dairy products of a
particular region or country.
 Fermented milk products constitute a vital component of the
human diet in many regions of the world specifically In the Indian
sub-continent.
 The production of these has been the monopoly of traditional
sweet makers and rural households.
 Total milk products is converted into different milk products such
as
 Ghee
 Dahi or Curd
 Khoa
 Paneer/Channa
 Rasbhari/ Lal Mohan
Methods of manufacture
Acid Coagulation & whey
Straining Paneer/
Channa Rasbhari
Straining Shrikand
Whole Lactic acid Dahi Whipping Lassi
Milk Fermentation Churning
Butter/ Makhan
Ghee Ghee
Residue
Separation Cream Churning Butter milk
Whole Butter
Milk Ghee Ghee
residue
Skim Milk
Acid Coagulation Paneer (non fat)
Lactic acid
Fermentation Dhai Straining whey
Chakka
Whole Milk Condensing Kulfi

Kheer
Khoa
Peda Burfi Lal Mohan

Microbial status: -
 Processing conditions are not hygienic at halwais, and

rural people so products are grossly contaminated
 Most of the products are sterile when fresh, it is the post-
processing contamination during storing that needs
vigorous checking
 Contamination increases spoilage organisms, lowering
keeping quality
 Entry of pathogens is possible depending on nature and
source of contamination
Microbial standards (BIS): -
Product SPC Coliforms Yeasts and

(cfu / g) (cfu /g) molds (cfu /g)
Khoa ------- 90 90
Burfi 30,000 ---- 10
Kulfi 2,50,000 100 -----
5,00,000 90 250
Paneer
Shrikand ----- 10 30
Canned rasogolla 500 Nil ----
Fermented milk ----- 10 100

products
Microbial quality of khoa
 Khoa is an indigenous milk product which is also called Kurauni

(in Nepali).
 It is produced in different regions for selling and domestic
consumption.
 Khoa is a product of great commercial importance as it forms an
important base material for the preparation of varieties of
indigenous milk based sweets throughout the country e.g. Peda,
Burfi etc
 Khoa has a low shelf-life ( 3 days in summer and 6-7 days in
winter).
 Khoa, can serve as a favorable medium for the growth of a
variety of microorganisms because of high moisture content
and good nutritive value
Microbial quality of khoa
 Rapid spoilage of khoa is attributed to the contamination

with molds from external sources
 Microflora: - Various groups of bacteria isolated from
Khoa include
 Acid producers,
 Proteolytic,
 Chromogenic,
 Lipolytic,
 Aerobic sporeformers,
 Yeasts and molds
Microflora in Khoa
Microbes Origin / Source
Micrococci : M. flavus, M. citreus, Soil, improperly cleaned
M. caseolyticus, M. luteus, M. roseus utensils
Sarcina Sp ,,
Staphylococci: S. aureus and Others Handlers
Bacillus: B. subtilis, B. cereus, Survival during manufacture and
B. megatherium, B. stearothermophilus multiplication during storage
Coliforms: E. coli, Enterobacter aerogenes Faecal contamination of the
product, directly or through
handlers, packaging etc
Streptococci: L.lactis ssp lactis, L.cremoris, Normal flora of milk
S.durans
Lactobacilli : L.acidophilus, L.bulgaricus ,,
Yeast: - Saccharomyces, candida Atmosphere
Torulopsis, Debaryomyces, Rhodotorula
Cryptococcus and others
Molds: - Pencillium, Aspergillus, ,,
Geatrichum, Syncephalestrum, Mucor,
Fusarium, Rhizopus, Cladosporium, Others
Microflora in Khoa
 Khoa contain higher population of yeast and molds than

burfi and pera.
 Among the bacteria the predominate ones in burfi and
pera are as follows.
BURFI PERA
1. 41.2% 22.3% Bacillus Sp
2. 31.8% 32.8% Staphylococcus
3. 23.5% 27.7% Micrococcus
Pathogenic Microflora in Khoa
 Coagulase positive staphylococci: - survival long storage of khoa

produces heat stable enterotoxins
 Enteropathogenic E.coli, Drug resistant coliforms, Enterococci –
Post processing contamination because they are heat labile.
 SalmonellaSp, Shigella Sp: - as pathogens are encountered in these
products
 Sources: -
 Faces
 Sewage
 Water supplies contaminated and faces
 Handlers
 Equipment
 Unhygienic environment
Factors influencing micro flora
in Khoa
 Factors during production, processing, handling,
storage and distribution are important.
 Major role is played by post processing factors as
most of the initial microbial population is killed by
severe heat treatment.
1. Quality of milk:
 Since the products are prepared mostly by halwais and
rural folk the milk obtained from rural areas
constitutes raw material,
 Such milk contains a heavy load of microbes including
heat resistant spore forming types and enterotoxins
producers.
in Khoa
1. Quality of milk
 The heat survival is due to heat coagulated proteins in
khoa act as insulators.
 Open pan method involves alternative heating and
cooling so thermal effect is reduced.
 The temperature of heating ranges between 62-68°C.
2. Production Hygiene:
 Utensils – as source of micrococci, bacilli and other
thermoduric
in Khoa
2. Production Hygiene:
 Soil and aerial contamination – environment where

direct air currents carrying soil and dirt are common, flies
contribute pathogens, air is source of aerobic spore
formers, yeasts and molds
 Handlers – very significant especially in pera where khoa
and powdered sugar are mixed with bare hands into
small flat ball called pera
 So it contains highest staphylococci (32.8%) since these
are associated with nails and skin.
in Khoa
3. Packaging:
 Paper boxes, parchment paper, aluminium foils,
polyethylene, Bamboo baskets and tree leaves.
 Contaminants: - Aerobic spore formers, Yeasts, Molds
and Others
 Prevention: -
 Chemical treatment of packaging materials with
antimicrobial agents
 Nitrogen atmosphere for packing
 Hot packing
in Khoa
4. Storage: -
 Condition varies RH 60-90%, temperature20-40°C.
 Longer the storage greater is the microbial population.
 Moisture also plays important role. Khoa has maximum
moisture than burfi and pera which have comparable
moisture
Defects
A. Rancidity: -
 Fat is degraded by microbial lipases.
 The organisms producing lipases are lipolytic bacteria, yeasts
/ molds.
 Both hydrolytic and ketonic rancidity might be produced
depending upon microbial activity.
 Control: - Storage less then 10°C, General hygiene
B. Stale and sour flavor:
 Stale is due proteolytic organisms and Sour due to acid
producing organisms
 Control: - minimize post-processing contamination, through
utensils and water supply
Defects
C. Mold growth:
 Imparts off flavors and produces mycotoxins.
Mold growth is favored by
 High moisture content in product
 Air leakage in package
 High humidity
 Sufficient aeration in storage room
Control
 Proper sanitation of dairy environment

 Controlled humidity and aeration in processing and
packaging rooms
 Moisture not more then prescribed limit
 Khoa stored in air tight package
 Use of potassium sorbate treatment of packaging material
Preservation of khoa:
 Shelf life 37°C 7 days
 23-24°C 10 days
 5-10°C 3 weeks
 Hot packaging – At 80-90°C gives keeping quality of 14-21 days at
37°C
 Drying of khoa – Dried khoa 90 days in air packed, 105 days in gas
packed at 16-30°C room temperature
 Heat sterilization – but affects sensory quality
 Addition of chemical preservative – BHA, potassium sorbate
 Addition of NISIN
 UV irradiation – for 90mts increase keeping quality from 5 to 25
days, but product developed oxidized flavor
 Lower temperature storage
 Addition of sugar – at the rate of 2% in milk, increase keeping
COAGULATED PRODUCTS (Paneer)
A .Paneer: -
 Moisture (not more than 70%), Milk Fat (not less than
50%)
 Made from buffalo milk (5% fat)
 Paneer made by coagulation with citric acid or tartaric
acid
 Paneer not much used in sweets more to make vegetable
dishes
 Microbiology: -Sources of contamination are air, water,
utensils, cutting knife, muslin cloth, person handling
COAGULATED PRODUCTS (Paneer)
 Microbial population vary due to

 Location of manufacture
 Extent exposure of product to air
 Extent exposure of product to temperature
 Period of storage
 Organisms: - coliforms
 Staphylococci (25% are coagulase positive)
 Yeast and molds
 TBC --, 35 x 10^5, 6000 X 10^5, average 6 X 10^5
COAGULATED PRODUCTS (Paneer/
Channa
B. Channa:
 Channa obtained by acid coagulation (lactic acid) of hot milk
 Channa made from cow milk only (4% fat)
 Channa is softer than paneer (not as firm) as it has no form
 Mainly used as base material for prepration of indian sweets like
Rasogolla, Rasmalai etc
 Spoilage is mainly due to thermoduric bacteria
 Micrococci – 45% predominant
 Sporeformers – 34%
 Non spore forming rods – 27%
 Common molds are –Penicillium, Asperigillus, Mucor,
Rhizopus,Fuserium etc.,
Defects
 Defects in paneer and chhana: - Rancidity, Mold growth

Microbiology of Rasbhari and
Lalmohan
 Very little information is available for the microbiological
quality of rasbhari and lalmohan.
 They are relatively safer due to its spongy and juicy

nature as a result of its soaking in sugar syrup.
 On long term storage can be contaminated with ,

Mesophiles, yeasts and molds
Microbiology of Ice cream and its
ingredients
Ice cream
 Frozen product that is most delicious and popular dairy

product.
 Ice cream is the frozen product obtained from :
 cow milk or buffalo milk or combination thereof or from
cream and/or other milk products,
 with or without addition of cane sugar, eggs, fruits, fruit
juices, preserved fruits, nuts, chocolate, edible flavors and
permitted food colors,
 It may contain permitted stabilizers, emulsifiers not
exceeding 0.5% by wt
 The mixture must be suitably heated before freezing.
Ice cream
 The product should contain not less than 10% milk fat, 3.5%
protein and 36.0% total solids.
 If any one of the aforesaid preparations contain fruits or nuts or
both, the content of milk fat may be proportionately reduced but
may not be less than 8% by wt.
 Starch may be added to a maximum extent of 5% with a
declaration to that effect on the label.
 Air cells are dispersed in a continuous liquid phase with
embedded ice crystals, the liquid phase also contains solidified fat
globules, milk proteins, insoluble salts, lactose crystals
 In some cases; stabilizers of colloidal suspension and sugars and
soluble salts in solution. So finished product constitutes a three
phase system
Suggested Microbiological
 SPC :< 50,000/g
standards
 Coliform : <10/gm
 S.aureus : <10/gm (coagulase +ve)
 E.coli(faecal type) : Absent/1g
 Salmonella : Absent in 25 g.
BIS STANDARDS:
 Weight (gm/lt) : Min 525
 Total solids(%wt) : Min 36.0
 Milk fat (%wt) : Min 10.0
 Acidity (%LA) : Max 0.25
 Sucrose(%wt) : Max 15.0
 Stabilizer/Emulsifier : Max 0.5
 SPC (per gm) : Not more than 2,50,000
 Coliform (per gm) : Not more than 90
 Phosphotase test : -ve
(British standards)
MBRT Test:
 2 ml sample
 7 ml Ringer’s solution.
 1 ml Methylene blue.
 Time taken
 Grade 1 - >4.5 hrs
 ,, 2 - 2.5 - 4.0 h
 ,, 3 - 0.5 - 2.0 h
 ,, 4 - 0 h
International Commission On
Microbiological Specification For Foods
(ICMSF) -
Ice cream standards
Limit per gram

Test Simple ice cream Complex Ice cream
m M m M
SPC 10^4 2.5X10^5 2.5 X 10^4 2.5X10^5
Coliforms 10 10^3 10^2 10^3
Staph. aureus 10 10^2 10 10^2
Salmonella spp. 0 0 0 0
History:
 Mention in Old Testament – milk & honey; Abraham & Isaac –

had frozen or chilled drinks;
 Chinese – mixed sugar and fruit juices to make an iced
sweet.;
 Ist century AD – Romans – ice used to chill mixture of honey
and fruit pulp or juice;
 In 1292 – Marco polo – details about sweet made from frozen
milk – sherbet ice;
 Introduced in Britain by Henrietta Maria, wife of Charles, I in
1630 brought her chefs with her.
History:
 Production – small scale until 1851 and Jacob fusel

opened the first ice cream factory in Baltimore, Maryland,
US in 1851.
 Hand worked ice cream freezer – invented in 1846 by
Nancy Johnson patented in 1848 W.G.Young.
 Major inventions in addition to mechanical refrigeration
(1878) are Homogenizer(1899)
 Invention of direct expansion ice cream freezer(1913) and
Continuous freezer (1929).
 The product is introduced in Europe, but industrially well
developed in America.
Sources of ingredients:
 Fat: Sweet cream; Plastic cream; Frozen cream; Unsalted
butter and Butter oil
 MSNF: Skim milk, Skim milk Powder, Condensed Skim milk
and Sweet cream butter milk
 Sources of both fat & MSNF: Whole milk, Whole milk
powder, Condensed Whole milk and evaporated milks
 Sweetening agents : Cane sugar or beet sugar (sucrose),Corn
sugar (9dextrose),
 Invert sugar (Glucose + Fructose),Corn syrup, Corn syrup
solids (dextrose + maltose), Saccharin and Aspartame
 Stabilizers: Gelatin, Sod. Alginate, Locust bean, Guar gum,
CMC (Carboxyl methyl cellulose), Pectin
Sources of ingredients:
 Emulsifiers: Glycerol mono stearate (GMS), Tween-80, Egg

yolk – (Active agent lecithin) Polyoxy ethyl glycol and
Sorbitol esters
 Flavors: Vanilla, Chocolate, Straw berry, Pine apple,
Lemon, Banana, Mango and Orange
 Fruits & Nuts: Apple, Banana, Mango, Pine apple, Grape,
Almond, Cashew nuts, Walnut and Ground nut.
 Colors: Yellow, Green, Pines etc.
Microbiology of Ice cream
ingredients
 Source of bacteria are of two types:
 Through ingredients
 Contamination at manufacture and handling

Microbiology of Ice cream
ingredients
 Ingredients added to ice cream mix before pasteurization
normally constitute little microbial contamination except
spores.
 However microbial analysis is very important in

establishing the quality of ingredients such as milk,
cream, other dairy products, chocolate, cocoa, eggs,
emulsifier and other food additives
 SPC
 Coliform
 Yeast & Moulds
 Thermoduric counts.
 Presence of higher bacteria in frozen dairy products and

ingredients are indications of improper sanitation and of
processing and handling conditions
Microbiological Analysis of
ingredients
1. Cream:-
 Bacterial contamination varies with raw milk used, separation
process,
 Inadequate refrigerated storage and Long distant transport
 Usually cream contains high bacterial counts than the milk
from which it is made and such cream may be a single chief
source of bacteria in ice cream.
 Best ice cream is made from sweet an un neutralized cream
which has not been subjected to microbial action.
 Cream with off flavor if used, will be reflected in the finished
product
ingredients
2. Butter or Butter oil (anhydrous milk fat)
 Tests –
 Yeasts & Moulds
 Mesophilic bacteria
 Coliforms
 Lipolytic bacteria.
 The important spoilage organism is Pseudomonas fragi which
causes unpleasant taste and taints.
 The spoilage is usually the result of chemical changes
producing rancid and other off flavors.
 Butter should preferably be stored at –18 to –20 c.
ingredients
3. Other Dairy products:
 Whole milk/Skim milk:
 High quality is desirable and it should be free from toxin
producing microorganisms.
• Raw milk should have SPC less than 2,00,000/ml
• Pasteurized milk SPC should be less than 30,000/ml

ingredients
3. Other Dairy products:
 Whole milk/Skim milk:
 Skim milk powder may contain number of Bacillus cereus,
though not a major health hazard but the number should
be as low as possible.
 Bacilli reduce methylene blue rapidly and can produce
clotting in milk; they grow at relatively low temperature
and in extreme cases cause spoilage of the mix.
ingredients
 Sweetened condensed milk (SCM) and Dried milk (DM):
 The bacterial standards intended for ice cream manufacturing
should
• SCM – SPC 500/ml
• DM - SPC 50,000/ml and Coliform 90/ml
 Condensed milk – Should be stored in manner that prevent

entrance of insects, bacteria and mold spores.
 Dry milk – should be stored in reduced humidity and in places

of free from insect & dust contamination.
ingredients
4. Vegetable fats –
 high temperature used in processes like refining and
deodorizing processes.
 They contain almost no moisture and therefore should
contain very few organisms indeed.
ingredients
5. Sugars and sweeteners:
 Granulated sugar and dextrose, should be almost sterile.
 Total count in any sugar should not exceed 200/g.
 The most common organism present in small number is
Yeasts.
 Sugar contains only small number of bacteria and is of little
importance as a source of bacteria in ice cream.
 Sugar syrups contain osmophilic yeasts and Moulds may
grow on the surface if contaminated.
ingredients
5. Sugars and sweeteners:
 Tests to be performed:
 Aerobic spore formers – Thermophiles, Mesophilic acid
producers
 Anaerobic thermophiles -H2S and non- H2S producers
 Yeasts and Moulds
 Coliforms.
 Thermophilic spores formers - not more than 125/10g - 150/10g
 Yeasts – not more than 10/10g
 Moulds – not more than 10/10g
 Flat sour spores - not more than 50/10g
ingredients
6. Stabilizers:
 Used to maintain smooth texture and body by preventing
formation of large ice crystals.
 Stabilizers should not present any problem, but gelatin as an

animal products, may be a hazard.
 The common organism in gelatin is aerobic spore former.

 Tests for stabilizers:
 SPC, Coliform, Yeast & Moulds, Aerobic spore formers which
indicate degree of unsanitary handling.
 Gelatin should be obtained from reputable supplies and store
as cool & dry.
 Gelatin
 SPC- not more than 10,000/g and
 Yeast & Moulds: not more than 100/g
 Stabilizers are kept dry and properly protected from dust and
dirt,
 The number of bacteria present tend to reduce as a result of
death of organisms.
ingredients
7. Emulsifiers:
 Very essential when butter, butter oil or vegetable fats are
used to provide surface active material in place of natural fat
globule membrane.
 Homogenization reduces fat globule size and increases
surface area which has to be protected.
 An emulsifier reduces the energy required to maintain the
integrity of fat globules.
 Assists in obtaining more uniform and large number of
smaller, air cells.
 Egg yolk is important source.
Tests for emulsifiers:
 SPC Coliform,
 Yeasts & Moulds,
 ß.haemolytic, streptococci,
 Salmonella
 Egg products – SPC should not exceed 10,000/g
ingredients
8. Fruits and Nuts:
 Important source of contamination in ice cream as they are
added after mix is pasteurized.
 Fruits – canned, fresh and frozen and should be of satisfactory in
microbiological standard particularly canned fruits.
 Fresh & frozen fruits – may containYeasts.
 Nuts may be infected with moulds.
 Coconuts may be contaminated with Salmonellae.
 Walnuts – Mould and parasitic infection
Tests for Fruits and Nuts:
 visual inspection,
 SPC, Coliform,
 ß-haemolytic streptococci,
 Yeasts & Moulds.
 Canned / fresh nuts are sterilized in ethylene oxide & CO2
in1:9 or boiling in sugar solution (50%) followed by drying at
2500C for few minutes.
 Coconuts should be heat treated when desiccated.
ingredients
9. Colors & Flavors:
 Careless handling – defects avoided by maintaining good
management control.
 Alcoholic colors add only a fewer bacteria. Flavors have
low counts due to
 presence of alcohol.
 Tests: Similar to fruits & nuts.
Suggests tests for raw
materials:
1.Milk - SPC, Coliform
2.Milk powder - SPC, Spore forming org.
3.Butter - SPC, Coliform, Ps. fragi, lipolytic, Y&M.
4.Cream - SPC, Coliform
5.Anhydrous milk fat - Coliform, lipolytic, Y&M.
6.Vegetable fats - Coliform, lipolytic, Y&M.
7.Sugar - SPC, Coliform,Y&M.
8.Stabilizer/emulsifier - SPC, Coliform
9.Fruits - SPC, Coliform,Y&M
10.nuts - SPC, coliforms,Y&M
11.Confectionary - SPC, Coliform,Y&M, Staphylococci
Processing of ice cream mix
 Heat treatment: A severe outbreak of typhoid in
Aberystwyth, Wales in 1947, caused by ice cream, led to the
introduction of heat treatment regulation ( Ice cream Heat
Treatment etc.
 Regulations, (1959) (SI 734) and amendment 1963 (SI 1083)
 Ice cream mixture must not be kept for more than 1h at any
temperature which exceeds 7.2°C (45°F) before being
pasteurized or sterilized.
Contamination at manufacture
and handling
1.Pasteurization:
Aim:
1. To destroy all pathogenic microorganisms
2. To reduce total number of bacteria
3. Through mixing of the ingredients, dispersion of
stabilizers.
Pasteurization temperature of 68.3°C /30 min or 71.1°C /10
min or 79.4°C /15 sec is followed
Pasteurization
 Higher pasteurization temperature required because of the

protection for the microorganisms by:
A. Sugar –
 Certain sugars retard the agglomerating action of heat on
protoplasmic colloids and as a result cells subjected to a given
heat treatment in concentrated sugar solutions are only
reversibly coagulated
 Where as in aqueous solution the protoplasm approaches
irreversible stage of coagulation
B. Increased milk solids present in the mix solids also protect
phosphatase against destruction.
Pasteurization
C. Concentration of the product interferes with heat penetration

 Ex: Micrococcus freudenreichii show more resistance to
destruction by heat in ice cream mix than in milk.
 M. tuberculosis is destroyed in 5 mts at 68.3°C that is why a six
fold safety margin has been given i.e 68.3 °C/30 min.
Micrococcus requires 79.4 °C /20 sec.
2. Sterilization:
 Temperature not less than 148.8°C held for at least 2 sec.
 The mix must then be cooled to not more than 7.2°C within
1.5 h of being heated and held till it is frozen.
3. Sources of contamination:
 Coolers & pipelines from pasteurizer equipment;
Condensates & pumps used for mixing after pasteurization.
 These two are important sources of coliforms
 Improperly closed vats may be a source of pathogens.
Homogenization
4. Homogenization:
 Homogenization pressure has little effect on microorganisms.
 Bacterial clumps are broken up and in maximum in total count
may be observed.
 Homogenizer must be kept clean and sanitized as it may
contribute to post pasteurization contamination of mix.
 If it is carelessly cleaned, it may be a very important source of
coliforms and other bacterial contamination.
 Homogenizer is a complex piece of equipment requiring
cleaning and sanitization.
Cooling
5. Cooling:
 Cools to below 4.4°C.
 Small increase in total count may result from
contaminants in the cooler.
 The sudden decrease in temperature has little effect on
the bacteria in the mix.
Ageing
6. Ageing:
 Changes are hydration of proteins and stabilizers; fats begin to
crystallize.
 Contaminants are introduced from the vats used for ageing.
16-24h at 4.4°C has little effect on the bacterial count of ice
cream mix.
 Under prolongation of storage can lead to the proliferation of
psychotropic organism with serious risk of mix spoilage
 Coliform bacteria also may increase in numbers in ice cream
mix held at 8°C & above which may give rise to false
conclusions relative to post pasteurization contamination.
Freezing
7. Freezing:
 Mix is frozen to the proper softness and whipped to the
desired overrun by the incorporation of air.
 Total bacterial count may be increased due to contamination
of equipment, disintegration of bacterial clumps during
operation.
 Few organism are killed by the freezing process itself due to
mechanical damage caused to the bacteria by the crystals
 Batch - -4 to –3°C /7 min
 Continuous - -6 to –5°C /24 sec
Freezing
 When the ice cream leaves the freezer the bacterial standard
is, as it were, “locked-in”, as very little change in the
bacteriological population will occur for several months,
provided the ice cream is kept properly frozen.
 Sal .typhosa – survive for 28 months at –20°C
 Paratyphoid & Brucella – 4 years at –23°C
 T.B organism - more than 30 months.
Packaging
8. Packaging:
 Packaging, molding and cutting of ice cream must be
carefully done to prevent contamination of ice cream.
 Small molds used for specialty ice cream are often or
serious source of contaminate because of their reuse
without adequate cleaning and sanitization.
 Hands of worker should be thoroughly washed and clothing
of worker should be fresh & clean.
Hardening and storage
9. Hardening & storage:

 Done at a temperature of -18 to –29°C.
 There is a gradual but not rapid decrease in total bacterial
number during storage.
 The water is crystalline state and is not available for microbial
metabolism.
Vendors
10. Vendors:
 Important source of contamination are dippers & scoops.
 Bacterial counts were high in scoop samples. Scoop dipped
frequently in water is of chief concern.
 Use of tap running water is good.
 Open top containers- chief contaminants for paper
containers from air/dust involve
 Micrococcus
 Alcaligenes
 Sarcina
 Bacillus and
 Achromobacter.
Microbial count in ice cream:
High microbial counts are due to:

 Poor quality ingredients.
 Improper pasteurization.
 Post pasteurization contamination.
 Improper ageing
 Unsanitary equipment
 Uninformed personal
 Unfiltered air used for over run.
Sources of coliforms:
 Leakage of raw milk with pasteurized mix because of leaky
valves etc.
 Pasteurized mix stored above 4.4°C. At 10°C there will be rapid
growth of coli forms.
 Prolonged storage of mix. i.e. above required period of aging (4
hrs.) Ingredients mostly fruits, nuts, colors, flavors.
 Absence of rapid sanitization programme.
 Improper sanitary practices by personnel involved in mix
processing.
 Foam acts as a good insulator so source of Coliform in vats.
 Large amount of sample will give more definite evidence than a
small one because of unequal distribution of bacteria, so
determination can be made on 2g amount of unmelted sample,
with deoxycholate lactose agar
Diseases through ice cream:
 Ice cream has been incriminated as a transmitter of pathogenic bacteria
particularly home made ice creams. The main source is being the raw
ingredients.
 The most common are
 Vibrio cholerae ,
 Ps. aeruginosa,
 M. tuberculosis,
 Sta. aureus,
 Sta.albus ,
 Sal. typhi (Typhoid fever),
 Sal. paratyphi (paratyphoid),
 Ent. faecalis,
 Cor. diptheridia (Diptheria),
 S. haemolyticus,
 St. pyogenes (Scarlet fever) and
Prevention and control:
1.Cold alone is not sufficient to kill pathogenic bacteria. Once it is
pasteurized it must not be exposed again to the possibility of
contamination by disease producing bacteria.
2. All ingredients should be free from pathogenic bacteria.
3. Careful operation of equipment, proper cleaning and disinfection of

plant at the end of each days operation, but also before it is use again.
4. Human contamination during handling of ingredients packaging

materials, produce processing & distribution.
5. The personal hygiene and habits of all in the factory – unwell - not
allow without full medical clearance.
Prevention and control:
6. Pet animals, such as dogs and cats, have no place in a food

production factory.
7. Use stream of running, pure, cold water for cleaning scoops and
other serving equipment.
8. Air used for over run should be filtered.
9. Ice cream should be protected from flies, rodents, which may

transmit diseases.
10. Maintaining proper hardening temperature storage &

temperature.
Hygiene:
Personnel :
 Educate about health hazards. Follow routine medical check up especially
for Skin diseases/sore wounds/abrasion.
 Wear clean overall and head covering
 Hand washing before entry into plant. Prohibition of smoking.
 Good personal habits
Plant:
Typical cleaning routine for freezer.
 Samples ice cream & mix is removed.
 Cool or warm 150°C - rinse.
 The freezer is then dismantled.
 Each part cleaned with detergent/disinfectant solution by hand scrubbing.
 All parts – rinsed in water & placed in a disinfectant solution before
reassembling.
 Disinfection carefully before it is used again.
Hygiene:
Other equipment:
1. The plant is thoroughly rinsed with warm water.
2. The plant is then dismantled.
3. Washing with detergents & physical methods of cleaning.
4. Re-assembling.
5. The plant is rinsed with clean water – temperature depression
the type of equipment.
6. Disinfecting – chemical solution – Sod Hypochlorite
7. Final rinsing, cleaning and then leaving the equipment to dry.
Precautions
Precautions taken to prevent contamination of products in small
size units where regulations are not followed:
 Hoppers should be refrigerated and the front of the freezer

should be insulated.
 The outlet of the freezer should be protected from droplet

infection
 The air used for over run should be filtered

Precautions
 The cans of mix should be around 4.4◦C when received
 The cans of mix should not be held too long and they should not
be left open
 The cans must be good in physical and sanitary conditions
 Facilities for washing and sanitizing should be available
 The public should not have access to the area around freezer
 The cans and containers should be stored in closed or self
closing units and finger contact should be avoided
 There should be adequate and convenient hand washing
facilities, including warm water, soap and clean towels.
Microbiology of Cheese
Introduction
 Food spoilage has been an important problem throughout

human history.
 Finding ways to overcome this problem was crucial as
communities became larger and individuals no longer grew their
own food.
 Some kind of system was needed to maintain the nutrient
content of various food stuffs for long periods of time and
prevent them from rotting and becoming inedible.
Introduction
 Early solutions to food spoilage

 Food spoilage is caused by the growth of microorganisms,
primarily bacteria and fungi, that convert nutrients into energy
which they use for their own growth.
 Depletion of the nutrient content of food as well as the secretion
of byproducts from this biochemical process, contribute to the
spoilage of food rendering it inedible.
 Ancient practices to extend shelf of food ; Salting, Drying,
Canning, Fermentation etc.
Introduction and History
 Cheese is an important product of fermentative lactic acid
bacteria.
 Particularly in the past, cheese was valued for its long shelf life.
 Due to its reduced water content, and acidic pH, bacterial growth
is severely inhibited.
 This causes cheese to spoil much more slowly than other milk
products.
 Art of cheese production has spread throughout Europe, each
country manufacturing many different types of cheeses.
 Made with biological materials milk, rennet and Mos, cheese is a
continually developing substrate, requiring time to reach
maturity and affected by the conditions under which it is
produced and stored
Introduction and History
 No real knowledge of the origins of cheese or cheese making.

 Preparation of cheese dates back many centuries to the time
when nomadic tribes of Mediterranean countries carried milk of
domesticated mammals in sacks, made from animal skins.
 If kept warm milk become sour and separated into curds and whey.
 If whey was drained from curds the latter could be dried to form
firm cheesy mass that could be eaten fresh or stored.
Types of Cheese
Types of Cheese
 The varieties may be grouped or classified into types according
to criteria such as
 Length of ageing
 Texture
 Methods of making/ ripening
 Fat content
 Animal milk
 Country or region of origin etc. - with these criteria either being
used singly or in combination
 The method most commonly and traditionally used is based on
moisture content, further narrowed down by fat content and
curing or ripening methods.
Types of Cheese
 General Classification of Cheese

 Soft Cheese
 Semi-hard Cheese
 Hard Cheese
Types of Cheese
 Hard Cheese (26-50% moisture)-ripened by bacteria
 Very hard (grating) e.g. Parmesan, Romano etc.
 Hard ( with eyes) e.g. Emmenthal etc.
 Hard without eyes e.g. Cheddar
 Semi-hard e.g. Gouda
 Bacterial surface-ripened cheese
Semi-soft (45-55%) e.g. Limburger etc.
 Internal Mould-ripened cheese
Semi Hard (42-52%) e.g. Roquefort, Bleu d’Auvergne etc.
 Soft cheese (48-55% moisture) Surface mold ripened e.g. Brie,
Camembert etc.
 Soft cheese (50-80% moisture)-Unripened e.g. Mozzarella, Ricotta
etc.
 Miscellaneous: White brined cheese e.g. Feta, Whey cheese e.g.
Ricotta, Processed cheese e.g. Many
Hard Cheese
 Hard pressed cheese made from firm, relatively dry curd
 Moisture percent range from 26-50%
 Hard cheese constitute a large group with 4 major types
 Ripened by bacteria and mature slowly over a period of 3-12
months
 In varieties like Cheddar acid is developed in the curd before
salting and pressing
 In varieties like Emmenthal acid is developed while the curd is
draining and being pressed before salting.
 Very hard cheese like Parmesan are made from very firm curd,
low moisture (26-34%), made from partly skimmed milk and
ripened by bacteria over a period of 1-2 years.
Hard Cheese
Characteristics of Hard Cheese
1. Low moisture content, achieved by size of cut, use of high
temperature for curd
2. Clean rind, or no rinds where film wrapped cheese are made
3. Method of ripening which proceeds evenly throughout the
whole cheese and carried out by enzymes derived from the
starter and non-starter bacteria and rennet.
4. Ripening is augmented and characteristics in Swiss cheese
by fermentation of lactate by propionic acid bacteria with
the production of CO2 which form eyes
Very Hard Cheese
 Grating varieties like Parmesan, Romano
 Very low fat (32%) and moisture contents (34%)
 Starter cultures of Thermophilic lactic acid bacteria
e.g. Streptococcus thermophilus with Lactobacillus helveticus,
Lactobacillus lactis ,Lactobacillus bulgaricus.
 Very high scald temperatures Long immersion in brine
 Long and slow maturation period
 During maturation, free fatty acids in the cheese increase slowly
producing sharp flavor of mature cheese
 Could be eaten fresh but usually stored for long periods, grated and
sold for culinary purposes.
Very Hard Cheese
 Parmesan Cheese
 Process
 Fat content of Milk 2%
 Very high scalding temperature (32-35◦C)
 Starter culture consists of Str. thermophilus and strong lactic acid
producers Lac. bulgaricus or similar heat-resistant Lactobacillus are
added (1%)
 After a short ripening period rennet (lipase rich rennet or rennet paste)
is added.
 Coagulum is cut into fine particles 3mm
 Curds and whey are stirred and heated in two stages 43◦C for 15min
and 54-58 ◦C for 15-30min
 Firm curd particles are settled for 10min and dipped out of whey in a
special cloth
Very Hard Cheese
 Parmesan Cheese
 Process
 Curd is allowed to drain for about 30 min prior pressing overnight.
 After removal from press cheese are held in moulds and kept in brining
chamber for 3 days at 15 ◦C for starter bacteria to continue ferment lactose.
 Cheese brined again at 26% NaCl at 8-10 ◦C for 14 days and dried for several
days at 15-20 ◦C and finally removed to curing room.
 Maturation 2 stages: first stage at 10 ◦C with RH 85% for 6-12 months. Turned
frequently to avoid spoilage of rind rubbed with vegetable oil to prevent mold
growth
 Second stage: matured for further 2-4 yrs
 Cheese coated with vegetable oil to prevent evaporation of moisture stored at
10-12 ◦C at 85-90% RH
 Proteolytic and lipolytic change produce strong flavors, cheese becomes drier
and could be grated
Very Hard Cheese
 Parmesan Cheese
 Microbiological changes
 During manufacturing starter bacteria increases
Str. thermophilus and Lac. bulgaricus and continue to multiply for few days
afterwards while the lactose is available in cheese.
 Non-starter bacteria from the raw milk have little time to grow before rennet
addition,
 Majority of Non-starter are killed by scalding temperature
 But extracellular enzymes of these bacteria contribute to proteolytic and
lipolytic changes in the cheese ripening.
 Low inhibition of cheese flora by salt during two weeks of ripening at 10 ◦C
while at 15-20◦C provides opportunity for growth of any bacteria survived until
(14 days) and which utilize the products of lactose fermentation
 Low moisture, low pH, low temperature long, slow ripening restricts further
microbial changes in cheese
Very Hard Cheese
 Parmesan Cheese
 Biochemical changes in protein, fats and carbohydrates which constitute
ripening are brought about by enzymes produced by bacteria in the original
milk or starter culture or by the enzymes from the rennet paste.
 Propionibacterium shermanii and P. freudenreichii were isolated from
Parmesan cheese; influenced by season, temperature of ripening on their
number.
 Abnormal counts of P. shermanii were associated with defects.
 Studies shown total numbers of bacteria in the cheese decreased markedly
during ripening from ≥10^7/g in fresh cheese to 10^5-10^4/g after 14 months.
 Decrease were more pronounced after the brine treatment or early in the
ripening period
 Psychotropic bacteria, coliform bacteria and Staph.aureus were regarded as
unlikely to cause problems in normal cheese and while propionic acid bacteria
occurred, their importance was not evident.
Very Hard Cheese
 Parmesan Cheese
 Lactic acid bacteria grew in relatively fresh cheese, but the starter starin
Lac.bulgaricus was eventually replaced Lac. casei as cheese matured.
 Spores of aerobic and anaerobic bacteria failed to germinate.
 Defects in parmesan cheese and other hard cheese arise from enzymatic
changes following bacterial contamination of Cheese milk
Associated defects
• Rancid
• Fruity
• Fermented flavors
 Surface defects: Not occur if proper care taken during ripening
 Heat treatment received by the curds and whey and treatment of brine
provide a substrate un conducive to growth of spoilage mos.
Hard Cheese without eyes
 Cheese in this group are not hard as grating cheese

 Made from whole milk and contains more fat (48%) and more moisture (39%)
 Starter used: Mesophilic lactic acid streptococci
 Lower cooking temperature used
 Acidity developed during the manufacturing process before the curd is salted
and pressed.
 Moisture is expelled from rennet coagulum by cutting, heating (cooking or
scalding) and development of acidity and following removal of whey
 Curd undergoes texturing in vat or in special devices
 Curd is milled and dry salt is added
 Curd is filled into moulds and pressed
 Conventional cheese are coated with paraffin wax to protect rind and
evaporation of moisture
 Matured in shelves at 15 ◦C and 88% RH
 Cheddar cheese
 Process and Microbiological changes
 Milk standardized to a certain ratio of casein to fat (C:F) (0.68-0.72) or fat to solid
not fat (F: SNF) (0.33-0.46)
 Pasteurization (71 ◦C for 15 s) cooled at 30 ◦C and delivered to making vats
 Inoculated with starter culture of Mesophilic lactic acid streptococci
Streptococcus cremoris, Streptococcus lactis
 After 20 min of inoculation rennet is added and 30-40min later coagulum is cut
into small cubes 5mm
 Acidity of whey maintained at 0.10-0.14% lactic acid
 Curd stirred in the whey and heated to 39-40 ◦C for 40-45 minutes (0.11-0.15%)
lactic acid
 Stirring continues until firm (dry) and suitable for pitching and separation of
whey (0.15-0.19%) lactic acid; final stage in curd making
 Maximum increase in Starter organisms takes place until this time
 Cheddar cheese
 Process and Microbiological changes
 After separation from whey the curd (0.20-0.22% lactic acid) is cheddared
either by hand or by cheddaring system.
 Starter bacteria continue to multiply and produce lactic acid at the rate of
approximately 0.10% every 20 min
 Whey continue to drain from the curd which becomes elastic, smooth and silky
 Eventually exhibits the characteristic ‘chicken breast’ texture(0.60-0.80%)
 The curd is then milled and salt(2%) is added and mixed thouroughly to ensure
even distribution
 Salted curd is delivered to cheese moulds and pressed hard(2-3 tonnes) for 15-
18 hrs
 The microflora of cheddar cheese:
• Starter bacteria: Streptococcus lactis, Streptococcus cremoris and
streptococcus lactis sub-sp. Diacetylactis
• Non strarter bacteria:
• May survive pasteurization or gained access as post pasteurization
contaminants
 Lactobacilli, Pediococci, Leuconostoc, Staphylococci, Micrococci, Group D
Streptococci and Gram Negative rods such as coliforms, Pseudomonads and
Achromobacter
• During cheese making the starter streptococci increase in numbers from
about 2.0 x 10^7 cfu/ml in the cheese milk 20min after inoculation to about 2
x 10^9cfu/ml in the curd at pressing.
• Greatest increase occurs in the first 3-3 1/2hr of the making process while the
curd is in contact with the whey.
• In period of salting (2-3h) the starter population is maintained or decline,
depending on the species and strains of starter bacteria.
 The micro flora of cheddar cheese

 Streptococcus lactis maintains its maximum population until
the cheese leaves the press while Str. Cremoris declines after
reaching its maximum.
 Survival depends on salt tolerance, Str. Lactis being more
resistant than Str. Cremoris .
 Fresh cheese curd contains between 10^8-10^9 viable starter
bacteria per gram, these die out during the early stages of
maturation at a rate dependent on the species; Str. Cremoris
tend to die out more rapidly than strains of Str. Lactis or Str.
Lactis sub-sp. diacetylactis

 Lactobacilli may be present in number ranging from 10 to 10^4/g in
the curd.
 Only lactic acid bacteria to multiply in maturing cheese (except
Pediococci) and reach levels of 10^6 to 10^8/g in 10 to 60 days,
maintain these numbers then decline after 4-6 months.
 E.g. Lac.casei, Lac.plantarum, the heterofermentative sps
Lac.brevis and Lac.buchneri and many unclassified Streptobacteria.
 Pediococci is less frequent but multiply at similar rate if present,
reach level of 10^7/g e.g. Pediococcus pentosaceus

 Leuconostoc may be present in curd in small number, usually 1-
100/g do not multiply and die out during ripening e.g. Leuconostoc
lactis, L.cremoris, L.dextranicus and L.mesenteroides.
 Group D streptococci, the numbers present may vary from being
completely absent to being present in small numbers of 10/g or
being present in high number of 10^4-10^6/g
• Group D streptococci die out a rate depending on the strain
sometime surviving for weeks and lasting for 3 months e.g.
Str.faecium, Str. bovis

 Micrococci occur in the curd at levels of 10^2-10^6/g.
-Depending partly on heat treatment of milk
- They slowly decrease in number during ripening, so after 6
months there is still 10-100/g of cheese.
- Coagulase-positive Staph.aureus strains which are potential
toxin-producers are destroyed by pasteurization.
- If lower heat treatment is used, this sub-lethal heating often
damages the cells, so if viable cells are present they grow on
favorable nutrient media
- Heat injured cells are unable to multiply on the unfavorable
acid (pH 5.2) and Salt (4.5%) substrate of cheddar curd.

 Micrococci :
-Heat resistant micrococci are commonly Micrococcus varians and other
unidentified micrococci.
-Other sps are M. lutes and M. lacticus
 Gram-negative rods:
- Killed by pasteurization
- Pseudomonads are only low-level post-pasteurization contaminants.
 Coliforms :
- Found much frequently, depending on hygiene of dairy personnel.
- Only non-starter bacteria to multiply during actual cheese making as
they are not inhibited by acid produced
- They die out rapidly in the cheese.

 Corynebacterium and aerobic spore-formers occur in small number in
the curd and survive for many months without multiplying or decreasing.
 During ripening controlled degradation of milk carbohydrate, protein
and butterfat occurs
 This is because of the action of enzymes from the milk, rennet, starter
and other micro flora and by chemical reaction like oxidation
 Yields a complex mixture of compounds which mature cheese with
required balance of flavor and aroma.
 Ripening: Three basic metabolic process, glycolysis, proteolysis, lipolysis
occurs
 Glycolysis: lactose lactic acid +acetic acid+CO2+diacetyl
 Starter streptococi and lactobacilli

 Ripening: Three basic metabolic process, glycolysis, proteolysis, lipolysis
occurs
 Lipolysis (weakly): Milk fat Free fatty acids (lactic acid bacteria)
 Proteolysis: Casein Peptides (Rennet)
Peptides + Amino acids (Starter Streptococci)

 Defects in Cheddar cheese: Rare if suitable starter used which
produce sufficient acidity at satisfactory level and which do not give
rise to fruity off-flavors or bitterness.
 Gassy curd, blowing of the film-wrap may arise from:
- Presence of Str.lactis sub-sp.diacetylactis in the starter which utilizes
citrate in the cheese milk producing CO2.
- Lac. casei also utilizes citrate, if present in the curd, grows during
ripening producing CO2, causes blowing of the pack at a latter stage
of maturation.
- High level of heterofermentative lactobacilli also produce CO2 during
glycolysis and can cause blowing of the flim-wrap, but high number
of lactobacilli must be present.
- Coliforms: if present in large number in cheese milk, multiply during
cheese making to produce gassy curd, which blow the film-wrap and
give fecal off-flavor
 Defects in Cheddar cheese:
 Rancidity:
 Heat resistant lipase from psychrotrophs (Pseudomonas) in raw milk if survive
at high level cause rancidity
 Rancidity occurs by hydrolyzing the milk fat triglycerides to release free fatty
acid
 These lipase remain active in cheese for more than 12 months so the levels of
FFA and intensity continue to rise
 Mould growth:
 Weak bodied cheese develop cracks and splits due to incorrect chees making
 May become infected with molds (Penicillium)
 Aerobic conditions cause splits allow their growth to penetrate the cheese
 Concentration of oxygen at the surface of flim-wrapped cheese should be too
low to support mold growth
Hard Cheese with Eyes
 Eye formation is the characteristics of certain hard cheese like

Swiss cheese, Emmenthal and Gruyere
 Eyes result from the production of CO2 in the cheese by
Propionibacterium
 The gas forms bubbles in the cheese which increase in size
depending on the rate of gas production and the physical
nature of the cheese.
 For these varieties of cheese it is essential to produce curd
which is sufficiently elastic for eye formation to occur.
 Propionic acid forms salts which contribute to the flavor of
these cheese.
Emmenthal cheese
 Named after the valley of the river Emmen –Swizerland
 Originated many centuries ago
 Traditional Swiss cheese – made in large wheels, 68-78 cm in diameter
and weight 70-80 Kg
 Nowadays rindless, rectangular blocks weighing about 40 kg
 The mature cheese has a pliable body with smooth texture, many large
holes , smooth and shiny and a characteristic sweet, nut like flavor
 Good quality raw milk is standardized to a fat content of 2.8-3.1%
 Milk warmed to 30-35◦C
 Starter culture: Thermophilic lactic acid bacteria such as
Streptococcus thermophilus (0.03-0.10%) with Lactobacillus lactis or L.
bulgaricus (0.20%) and Propionibacterium (1-1.25ml per 1000litres)
 The starter culture must be heat resistant and able to grow at relatively
high temperatures to survive the cooking process and continue to
grow during processing
Emmenthal cheese
 Rennet extract is added and 25-30min later coagulum is cut into
strips, then cubes and finally into particles about 3mm.
 Temperature of curd is raised to 45◦C at a rate of 1 ◦C every 2 min
and 53-57 ◦C at the rate of 1 ◦C /min.; temperature is critical to kill
non-thermoduric bacteria.
 Total heating time 45min, until curd is sufficiently firm
 Collected in large, coarse dipping cloth and then kept in the mould
of wood or stainlesssteel for draining table with pressing device ;
pressed for 20-24h and turned at intervals to promote uniform
drainage.
 After cooling of curd the starter bacteria multiply and produce the
acidity needed to cause the curd particles to mat together and form
elastic mass.
 Salting at 10 ◦C for 1-2 days, turned and dry salt is sprinkled on
surface removed from the mould immersed for 2-3 days in brine
containing 23% NaCl at 8-12 ◦C
Emmenthal cheese
 After removal from brine the cheese is placed on clean base broad
and held in chilled room 10-12 ◦C 80-85% RH for 8-10 days.
 Each day brushed and wiped with a cloth dipped in salt water
turned and dry salted.
 Maturation: Cheese is moved to warm room (18-23 ◦C 80-85% RH)
to encourage development of eye forming bacteria.
 Turned at intervals during 4-8 weeks, brushed, washed with brine
to prevent mold growth.
 Cheese acquires a slightly round shape, when eye development is
considered satisfactory, removed to curing room (13 ◦C and 80-
85% RH); washed, turned at intervals throughout the remainder
period (3-9 months)
 6 months are necessary for cheese to develop characteristics flavor.
Emmenthal cheese
 Propionibacteria bacteria and it activity
 Propionibacteria: development of the characteristic flavour

and eye formation in swiss cheeese
 Ferments lactic acid, carbohydrates and
polyhydroxyalcohols to propionic acid , acetic acid and CO2
 Growth of propionibacteria is stimulated in presence of
starter culture and more propionic and CO2 are produced
with improved flavour in the cheese
Emmenthal cheese
 Role of Micro flora in processing and ripening
 During curd making the no. of Str. Thermophilus increase slowly and
steadily but rapid at higher temperature towards end of cooking.
 Lac. Bulgaricus may decline in number
 Low cooking temperature may allow survival of an unwanted flora,
leading to defects.
 Draining and pressing (20 ◦C for 20-24h)control moisture of curd and
promote acid production assisting to the curd to mat together.
 At this period temperature of cheese falls and growth of starter and
acid production increases
 Cooling rapid on surface and thermophilic lactic acid bacteria began to
multiply there in first few hours of pressing.
 Str. Thermophilus multiplies first producing lactic acid at the periphery
within 2-3 h maximum no 10^8/g in the centre of cheese and 10^9/g at
the periphery about 3h later
Emmenthal cheese
 Lactobacilli multiply later
 Lac.bulgaricus after 4-5h reaching maximum in 20h; count being 10
times less at the center of cheese.
 Lactic acid fermentation rapid in periphery, hence lactose from the
center diffuses towards periphery where it is utilized, so less lactose
is available in center when temperature is eventually suitable for
lactic acid bacteria to metabolize.
 After removal from press (24h) no lactose remains, more lactate at
periphery than in center. This affects development of
propionibacteria and formation of CO2 and volatile fatty acids
 pH from pressed cheese 5-2-5-5 if higher then there may be defects
in eye formation and flavor
 If starter bacteria is inhibited by antibiotic residues in m ilk early
browning may occur.
Emmenthal cheese
 Propionibacterium:
 Inhibited by relatively low salt concentration and must develop
while the salt is still largely confined to periphery of the cheese.
 Excessive eye formation indicate low salt level in cheese
 When 7-10 day old cheese is moved to warm room
Propionobateria rapidly develop 10^9/g at 4-8 weeks and major
flora for the rest of ripening period.
Emmenthal cheese(Defects)
 Lipolytic Off Flavours

 Storage of milk at refrigeration for several days may create
problems
 Due to the growth of lipolytic psychrotrophic bacteria
 Early blowing
 Inhibition of starter bacteria by antibiotics in the milk or too low
heat treatment temperature and/or low cooking temperatures
can lead to the growth of unwanted bacteria such as Aerobacter
aerogens causing gas production in the curd whilst in the press
 Secondary fermentation
 Following the desired eye formation by propionibacteria a
subsequent secondary fermentation may occur
 Secondary fermentation(cont..)
 Results more eye production and the development of cracks in
the cheese
 This is because of late formation of CO2 from decarboxylation of
amino acids by enterococci or stimulation of propionibacteria by
free amino acids formed by the enterococci or proteolytic activity
of some lactic acid bacteria
 Lactobacillus helviticus is considered to inhibit late fermentation
 Late Blowing
 Abnormal production of gas (CO2) during ripening by lactate
fermenting clostridia
 These bacteria produce large amounts of butyric acid and CO2
and can cause severe spoilage of swiss cheese
 Excessive Eye formation or oversetting

 Large number of small eyes can result from excessive growth
of propionibacteria
 This fault usually indicates a lack of salt or acid in the cheese
 Lack of eye formation
 Propionibacteria are very sensitive to salt and acid
 High amounts of these can inhibit their growth
 Such high levels of salt or acid also cause loss of elasticity in
the curd which then cracks instead of stretching when eyes
are being formed
 This is known as the glass or glasler defect
 Various off flavours

 May develop if unwanted microbes grow
 All the gas producing organisms form products which cause
off-flavours such as yeasty, fermented or unclean or putrid
 Rusty spot
 Small rust colored spots may seen sometimes
 Usually caused by P. rubrum
Semi-Hard Cheese
 Cheese in this group vary considerably, demonstrate diverse
roles of lactic acid bacteria in cheese making, ripening and
flavor development
 E.g. Caerphilly, Lancashire, Edam and Gouda etc.
• Caerphilly: Fresh, acid curd
• Lancashire: Crumbly flavorful, mature
• Edam and Gouda: Closed textured, mild washed curd cheese
 All varieties are prepared from whole milk or low fat milk
 Starter culture: Mesophilic lactic acid Streptococci
 Rennet curds are coarsely cut and lightly scalded, thus retains
high level of moisture and lactose
Semi-Hard Cheese
 Specific methods are used subsequently to control the growth
of starter bacteria and development of acidity in different
curds.
 Differences in methods governs pH of young cheese and
accounts for widely different types of cheese within this group.
 Caerphilly and Lancashire : Growth of starter bacteria is
encouraged during curd making and pH of young cheese is low
(5.0-5.2) acid curds produced and they have crumbly texture.
 Edam and Gouda: Acid production is restricted by removing a
portion of whey during stirring and replacing it with water. It
reduces water content of young cheese, pH is high (5.3-5.4),
the body is plastic and texture is close
Semi-Hard Cheese
 E.g. Gouda and Edam
 Originated in Holland , most important type of cheese and
manufacture is well mechanized.
 Gouda is made in the form of small wheels weighing 3.5-25kg.
 Edam is made in small balls weighing 2kg, red waxed.
 Both varieties are sweet curd, renneted cheese made by the
system of washing curds with water.
 High pH of both curds (5.3-5.4) and absence of lactic acid to
suppress spoilage bacteria, emphasize the importance of using
milk of good bacteriological quality for making cheese.
 Bactofugation of milk for Gouda improves the cheese quality
by removing a considerable number of clostridia spores
present
Semi-Hard Cheese
 E.g. Gouda and Edam

 Equipment's: Cheese making vessels, ancillary tools, cheese
moulds and presses, brine tanks and cheese stores.
 Process and Microbial changes:
 Milk is standardized and pasteurized and cooled to 30-31◦C
 Calcium chloride (up to 0.02%), Sodium nitrate (up to 0.02%)
 Coloring may be added before milk is inoculated with 0.5% of
starter culture of Mesophilic lactic streptococci ( Str. cremoris,
Str. lactis)
 Rennet (30ml per 100 liters of milk) is added and 30 min later
coagulum is cut into large particles (10mm cubes).
Semi-Hard Cheese
 E.g. Gouda and Edam
 Process and Microbial changes
 These are lightly scalded by removing some whey and replacing
it with water at 50-60 ◦C; this raises the vat temperature to 36-37
◦C in around 30 min.
 Amount of the water added is equivalent to 25% of the original
milk and this dilution reduces the lactose content of the curd and
controls the growth of the starter bacteria.
 After draining off the whey , the high moisture, high pH (5.3-5.4)
curds are filled into moulds and presses lightly .
 The cheese curds are then salted in brine (21-36% NaCl) for 24-
48h dried to form a coat .
 Ripened for about 6 weeks (12-15 ◦C and 85-90% RH)
 They are waxed or film wrapped before sale.
Semi-Hard Cheese
 Changes during ripening are similar to other hard-pressed
cheese-breakdown of casein by enzymes from rennet, and by
protease from starter bacteria with further degrade the
intermediate products of casein hydrolysis (proteoses and
peptones).
 But this is restricted by high pH of young curd (5.3 at 24h).
 Total count of cheese increases rapidly at the start of ripening
and decreases slowly.
 Str. Cremoris is present in large number during first 10 days and
quickly disappears
 As the starter bacteria grow and the pH of cheese decreases (5.1
at 3 days), casein hydrolysis intensifies
Semi-Hard Cheese
 Particular strains of lactic acid bacteria vary in their ability to
break down the products of casein hydrolysis and their effect
on the quality and micro flora of Edam cheese.
 Gouda with its slightly lower moisture content can be
matured for longer than Edam.
 It allows more protein breakdown and more flavor.
 A few small eyes produced by propionic acid bacteria are
permissible in Gouda but regarded as defect in Edam.
Semi-Hard Cheese
(Defects)
 Gouda and Edam cheese suffer may defects which spoil hard
cheese such as
 Mould growth (mainly by Penicillium and Aspergillus sps.)
 Gassiness
 Development of off-flavors and
 Bitterness
Bacterial Surface-
Ripened Cheese
 Semi-soft Cheese (e.g. Limburger, Port du Salut)
 This group of cheese contains a number of varieties which
are ripened largely from outside by the action of surface
flora.
 All varieties made from sweet, rennet curds and are
ripened to some extent by proteolysis from starter
bacteria.
 Characteristics brownish-red surface growth of Bre. lines
makes contribution to the ripening of some varieties and
affects the flavor of all of them
Bacterial Surface-
Ripened Cheese
 Semi-soft Cheese (e.g. Limburger)
 Process
 Made from Pasteurized milk with small amounts of starter
culture of lactic acid streptococci
 Starter culture may be Str. lactis alone or Str. lactis and Str.
thermophilus and amount may vary from 0.1-0.2% for
Limburger to 0.8% for Port. du Salut.
 Rennet (22ml per 100 liters) is added at 30-32 ◦C , the
coagulum is cut in 30-40min, and the curd particles are
heated in the whey.
 Moisture content of different varieties is controlled by the
particle size and the temperature of the scald 35 ◦C for
Limburger
Bacterial Surface-
Ripened Cheese
 Process
 After removal of whey, the curd is filled into suitable cheese
moulds followers and weights are placed on top of curd to
consolidate it , turned frequently for several days.
 Immersed in the brine (23% NaCl) at 10 ◦C for 24h or dry
salted on surface for 1-2 days and wiped with cloths soaked
in brine.
 Cheese are now placed on shelves in the ripening cellar.
 Within few days characteristics brownish-red slime develops
on the surface and this is uniformly smeared over whole
surface to distribute the bacteria uniformly.
Bacterial Surface-
Ripened Cheese
 Process
 Size varies in these types of cheese
 Important relationship between depth of cheese and
surface diameter is important, since too large diameter or
too great a depth can accelerate or delay ripening, cheese
could over ripe and bitter near the surface long before the
interior is ripe.
 Role of Micro flora in Processing and Ripening
 Starter bacteria:
• During manufacturing the lactic acid streptococci grow
rapidly, and development of acidity continues while the
curd is draining in the moulds
Bacterial Surface-
Ripened Cheese
 Starter bacteria:
• During manufacturing the lactic acid streptococci grow rapidly,
and development of acidity continues while the curd is draining
in the moulds.
• After 24h the pH may be about 5.0 but this varies with different
varieties.
• Rate of acid development governs the rate of whey drainage
from the curd and thus final moisture content of the cheese.
• Affected by the numbers, activity of acid producing bacteria,
and by the room temperature.
• Rapid acid production, leading to a dry, over-firm curd or slow,
inadequate drainage leading to a wet, sour curd, result in cheese
which don’t ripen satisfactorily.
Bacterial Surface-
Ripened Cheese
 Surface flora:
• Within 2-3 days of manufacture, a whitish growth of aerobic,
salt tolerant yeast and Geotrichum candidum appears on the
surface of the cheese.
• Some days later when pH has been increased by their
metabolism, the brownish-red growth of Bre. lines appears
and surface becomes slimy.
• Brevibacterium linens is very actively proteolytic and the
extent to which its growth is allowed to develop depends on
the variety of the cheese.
• It is greatest in Limburger where cheese may resemble the
consistency of warm butter.
Bacterial Surface-
Ripened Cheese
 Ripening:
• In these cheese as the starter bacteria die out, the curd undergoes
varying of proteolytic and lipolytic changes associated with
enzymes from the starter bacteria and rennet.
• Function of the surface organisms esp. Bre. linens , is to
contribute, in a greater or lesser degree to the flavor .
• Their growth and contribution to cheese flavor is governed by
1. Moisture content of the curd, drier varieties such as Bel Paesa
and Monterey show less surface ripening and low flavor intensity
2. Area of cheese exposed during ripening, cheeses may be piled on
top of one another to prevent excessive growth of Bre. linens.
3. Temperature and duration of ripening
4. Removal of the surface growth
Bacterial Surface-
Ripened Cheese
 In Limburger the surface growth is responsible for many of the
changes in ripening.
 G. candidum utilizes the lactate formed and increases the pH at
the cheese surface to enable Bre. Linens to grow.
 The difference in pH from surface (6-7) to centre (5.4-5.7) suggest
that neutralization of the acid is associated with the products of
the surface flora rather than casein hydrolysis within the cheese.
Bacterial Surface-
Ripened Cheese
(Defects)
 Semi-soft Cheese
 Moulds:
-Most serious defects arising in bacterial surface ripened cheese is
failure to develop.
- Occurs if the surface of the cheese is too dry, or the humidity of
curing room is too low.
- Slow development of the smear allows unwanted moulds to grow
- Cladosporium, Penicillium, Aspergillus etc
 Early blowing:
-If good quality pasteurized milk is not used all varieties may suffer
early gas formation by coliform organisms during draining and salting.
- This defect may vary in intensity from a few, small pin-holes to a
Bacterial Surface-
Ripened Cheese
(Defects)
 Semi-soft Cheese
 Late Blowing
- If curd after manufacture does not have a low enough pH (below
5.3) or high enough salt content the cheese are subject to late gas
formation by the growth of clostridia
 Over-acidity:
- Excessive development of acidity by the starter bacteria during
manufacture may cause the pH of the curd to fall too rapidly or too far
(4.8-4.7).
Internal Mould-
Ripened Cheese
 E.g. Gorgonzola, Roquefort and Stilton
 Made from high acid, semi-soft curd and the process involves slow
acid development during long draining period.
 These cheese are not pressed, instead curd is allowed to consolidate
under their own weight.
 Salt may be added to the drained, acid curd before it is placed in the
cheese moulds (Stilton) or may be rubbed into surface of those
cheese where acid development takes place in the moulds
(Gorgonzola, Roquefort ).
 Initially blue-viened cheese may undergo some proteolysis from the
starter bacteria and rennet enzymes
 When they are pierced to admit air the blue-green moulds,
Penicillium roqueforti or P. glaucum spread through the cheese and
brings proteolytic and liplytic changes responsible for ripening and
Internal Mould-
Ripened Cheese
 Cheese Making
 Milk : For blue–veined cheese , high-fat milk desirable as lipolysis
plays an important part in flavor development.
 Sheep’s milk extensively in France and mandatory for Roquefort and
Stilton but Gorgonzola made from cow’s milk and cream added if
fat content is low.
 Most of the milk used is raw, as milk lipase aids fat hydrolysis in
ripening but now pasteurized and homogenized milk is been used to
overcome defects of raw milk of poor bacteriological quality.
 This process helps to form a smooth curd with maximum moisture
retention, increases the surface area of the fat, promoting the
lipolytic action of P. roquefortii and accelerating typical flavor
development.
Internal Mould-Ripened Cheese
 Cheese Making
 Starters: Mesophilic lactic acid bacteria
 Growth and acid production of starter are essential for adequate
draining of curd.
 Str. Lactis sub-sp. diacetylactis and Leuconostoc
 Moulds: Blue-veined cheeses are characterized in appearance and
flavor by growth and development of blue mold P. roquefortii.
 Cultures of P. roquefortii in liquid suspension are introduced
deliberately either to the milk before rennet is added or by spraying
on to the curd with the salt.
 Cheese are pierced several times, during ripening to admit air to
enable the mold to grow and metabolize
 Proteolytic and lipolytic activity within the cheese gives the cheese
their piquant character and flavor
 Cheese Making
 Process:
 Closely associated with particular geographical localities for e.g.
- Stilton ; Made in farm house in the Vale of Belvoir in Leicestershire.
- Gorgonzola; Milan Italy
- Roquefort: 8th century Province Aveyron France
 Basic principle of high acid development over a long slow drainage period: pH
of 4.5-4.7 is reached in 24h.
 Slow acid development controls the expulsion of moisture from rennet curd ,
to produce a cheese with soft velvety body, flaky texture, low pH and
moisture 48-50%
 Starter and rennet are added to the milk , after cutting, syneresis of the curd
is brought about by acid development only.
 Curd are not scalded during processing and not pressed mechanically.
 Curds filled into moulds, frequent turning ensures adequate drainage and
 Role of Micro flora in processing and ripening:
 Slow production of acid by the lactic acid bacteria makes it essential to use
good bacteriological quality.
 Balance between acid production and moisture expulsion is vital any
acceleration or hindrance of acid development affects the moisture level in
the curd.
 Salting of curd before moulding reduces moisture content or applied to
cheese helps to firm and harden the surface.
 During salting the no. of starter bacteria are high 10^9 cfu/g but low pH and
increasing concentration of salt in the moisture of cheese inhibit these
organisms
 But after 2-3 weeks few viable cells are found, no other bacteria are found
during ripening.
 Actual mechanism of starter bacteria for ripening is not known, possibly due
to intracellular enzymes released and rennet bring proteolysis of casein in
the early stages of ripening and before the mold (P. roquefortii) grows.
Role of Micro flora in processing and ripening:
 Characteristics process in the manufacture of the cheese is pricking,
piercing or stabbing the cheese to admit air, possibly mold spore, to the
interior.
 Roquefort and Gorgonzola: Pierced at 2-3 weeks old
 Stilton: 5-6 weeks old
 A t the temperature (10-13°C) and humidity 96% of curing room P.
roquefortii
appears within 8-10 days.
 Mould ramifies through fissures in the cheese and along the lines of
stabbings maximum in 30-90 days.
 Maximum growth at the centre of the cheese where initially the salt
concentration of salt is lower.
 Cheese are then wrapped in foil or moved to a cooler curing room to slow
down ripening during remaining maturation process.
 P. roquefortii is used as ripening agent due to its tolerance of salt and low
concentration of oxygen than other sps., added in huge number.
 After ripening the oxygen content is reduced and CO2 is increased (P.
roquefortii withstand them better than other species).
 Casein is hydrolyzed after mold develops and body of cheese becomes softer
and increase in amount of amino nitrogen (marked during 1st month of
ripening) but slowly continues throughout the life of cheese.
 Proteinase activity is over a range pH 5.3-7 and optimum pH 6.
 Ripening:
-pH of cheese rises from initial minimum of 4.7-4.5 at 24h to maximum of 6.0-
6.24 at 2-3 months; as a result of utilization of lactic acid by P. roquefortii .
- Subsequent fall in pH occurs when butyric, caporic, caprilllic and capric acid or
if other higher fatty acids liberated from fat by lipases produced by mold.
- -Lipases are active at pH, temperature and salt concentration found in
ripening cheese.
 Coating:
- Throughout the making process and until a satisfactory rind or coat has
formed these cheese are vulnerable to infection by air-borne molds and flies.
- Formation of relatively smooth, impervious coat is necessary for protection
of cheese during ripening, coat may vary with type of cheese.
- Surface flora affected by pH, moisture, concentration of NaCl.
- Salt tolerant yeast develop first increasing the pH , so other mos can grow.
- Reddish-brown growth of Bre.linens may appear, and cheese acquires slimy
type of surface (e.g. American Blue)
- While cheese like Stilton, have drier surface
- Yeast, lactobacilli are found regularly, with aerobic spore former, pigmented
cocci and coli form occur irregularly in variable numbers.
- Type of coat is often characteristic of particular curing room.
Defects in Internal Mould-
Ripened Cheese
1. Gassiness:
- Gassiness is seldom troublesome due to low pH and high salt concentration
in the young cheese, combine with fairly low curing temperature provide
unsuitable conditions for growth of spoilage bacteria.
- Volatile fatty acids can be produced quite early during ripening, but if milk
is homogenized, are inhibitory to many bacteria and may assist in the
control of bacterial spoilage.
2. Failure to blue:
- Failure of P. roquefortii to develop properly and consequent low levels of
proteolysis and lipolysis, produces a cheese which is too firm and lacks
flavor.
- Poor mold growth results due to excessive development of acidity in the
curd, leading to too close texture in the cheese or dry curd.
- Low humidity in making room also cause dry curd and affect acid
development.
- Poor color of mold arise from species of Penicillium used or possible lack of
iron in milk.
Defects in Internal Mould-
Ripened Cheese
3. Over-bluing:
-Excessive mold growth may occur if cheese are pricked heavily and proportion
of O2 to CO2 is upset.
-A good blue color is achieved but volatile acids are used for the mold
metabolism and subtle flavor is not produced.
-Worst condition is musty, unclean flavor development.
4. Surface defects: Causes considerable losses.
-A black mold usually Caldosporium may grow on surface and stab-holes causing
musty flavor.
-Application of paraffin wax or Cryovac or Al-foil after satisfactory mold growth
is sometimes used to protect cheese.
5. Browning: Common defect associated with high pH (8.0-8.5) in cheese.
6. Slip-coat: In Stilton cheese and soft edges in Roquefort are defects associated
with moisture under the rind of the cheese.
Strong Proteolytic organisms like Proteus vulgaris and Str. liquefaciens were
found to be associated with slip coat other factors may be involved.
Soft cheese
 Constitute a large group of varieties characterized by moisture
content of (48-55%) to high (55-80%) and consequent
perishability.
 Made from cream, whole milk, or whole milk standardized
with cream or skim-milk.
 Curds may be formed by rennet and acid and if starters are
used they are mesophilic lactic streptococci.
 Curd receive little or no cutting and little scalding.
 Cheese are not pressed, any necessary drainage is done by
gravity.
 High acidity may develop in certain varieties.
 Cheese made in small cylindrical, rectangular or logs shapes.
 Some varieties are consumed fresh and have clean lactic acid
flavor with little break down of protein or fat.
Soft cheese
 Types of soft cheese

 Soft cheese (48-55% moisture) Surface Mold-Ripened e.g.
Brie, Camembert etc.
 Surface Smear-Ripened Soft cheese e.g. Romadour
 Soft cheese (50-80% moisture)-Unripened e.g. Mozzarella,
Ricotta etc.
Soft cheese
 Includes many varieties of soft cheese made in France.
 These are full fat soft cheese with 45-50% FDM (Fat in dry
matter) and maximum moisture of 55%.
 Made from cow’s milk but sometime goat’s milk may be used.
 Cheese prepared from whole milk with addition of starter and
rennet to form a medium acid, soft curd.
 Curd is transferred without cooking to perforated moulds and
drainers.
 Frequent turning of moulds help to achieve adequate drainage
and subsequent ripening and gives cheese a sufficient dry
surface for yeast and mold establishment.
Soft cheese
 Strong Proteolytic enzymes produced by the mold P.
camemberti diffuse into the curd converting it to characteristic
soft, smooth condition during a relatively short ripening
period.
 Surface mold ripened cheese are made in small sizes to
provide the maximum surface for mould growth, and optimum
distance for diffusion of their enzymes.
 Mature cheese deteriorate rapidly , and in absence of
refrigerated storage must be marketed quickly.
Soft cheese
 Soft cheese :Surface Mold-Ripened
 Camembert
 Most important of French soft cheese.
 Making room should be kept warm (22-25°C)
 Made from fresh milk, sometimes raw milk is used but
generally heat treated (72 °C for 16s) cooled (33-34°C) and
delivered to making vats.
 Culture (1.5-2.5%) of lactic acid streptococci is added and
allowed to ripen the milk for 1h before rennet (16-17ml per 100
ltrs) is added.
 Coagulation begins in 25min, curd should be firm enough for
distribution in 70-75min from renneting.
 Curd is ladled into perforated cylindrical moulds, standing on
salts on a draining table.
Soft cheese
 Camembert :
 Drainage proceeds for about 6h and during this time, the curd should have
shrunk to half its original height and acidity of the whey about (0.60-0.70%)
lactic acid.
 Cheese are then turned and left overnight for draining (min. 22°C)
 Changes during Processing and ripening
 After removal from the moulds moisture content is 60%.
 Temperature and humidity of making room helps to achieve the balance
between increasing acidity and decreasing moisture content for
development of surface flora.
 Low temperature impair acid development and drainage and high
temperature accelerate acid development and cause excessive drainage.
 Dry salting after 24h at 18-20°C helps to dry the surface of cheese further
and to control the type of organisms growing on it.
Soft cheese
 Camembert :
 Cheese are sprayed with P. camemberti culture and 24h later transferred to
ripening room.
 During this time starter bacteria have multiplied and risen to very high
number in the curd but after few days at start of ripening their number
begin to decline due to low pH (4.7-4.9) , shortage of lactose and high salt
concentration.
 Species of lactobacilli may be found during ripening , but their effect is
considered insignificant.
 Maturation first (10-14 days) temperature of 11-14°C and RH 85-90%
maintained ; growth of film yeast (Mycoderma) and Geotrichum appear on
surface (withstand high acid and high salt content ).
 Few days later P. camemberti begins to develop and spreads the whole
surface
 Maximum development in 10-12 days, when acidity at the surface has been
further reduced by yeast and mold the red and brown spots of Bre.linens
appear.
Soft cheese

 Camembert : Microbiological changes
 Complex changes occur
 Yeast grows first may ferment residual lactose at the cheese surface and
reduce acidity, but their growth should not be excessive, otherwise spores of
P. camemberti may have difficulty in penetrating the yeast mat and
establishing themselves.
 Yeast restricted to certain extent by salt content of the cheese surface and by
regulating temperature and humidity; surface is fairly dry.
 Luxuriant snow-white growth of P. camemberti spreads over the whole
surface of the cheese and changes body and flavor of cheese.
Soft cheese
 Camembert : Microbiological changes
 Efficient control if atmospheric conditions in repining room leads to its
proper development.-
-Too low temperature: slow mold growth and ripening delayed.
- High temperature lead to excessive mold growth and strong flavors
– Low humidity: cheese surface become too dry, prevents yeast and molds
from growing and wild sps like green spored penicillia grows.
- High humidity: Movement of air inadequate, evaporation of moisture from
cheese surface is insufficient, allows excessive growth of film yeast
Geotrichum and Mucor sps, and P. camemberti inhibited.
 After 10-14 days after ripening begins, the edges of cheese soften at this
stage the cheese is wrapped and boxed.
 By time they reach to consumer 2-3 wks later this softening will have
extended to the centre of the cheese, fine flavor will developed and
ripening will be completed.
Soft cheese
 Camembert : Defects
 Excessive proteolysis and Strong flavor-initial high moisture content or
high temperature in ripening room
 Cheese with dry curd or dry surfaces-low humidity
 Over salting or under salting
 Cheese with wet surfaces; risk of excessive growth of Bre.linens. Results in
consequent surface smear rather than surface mold.
 Early gas: during curd draining, if raw milk is used.
 Late gas: Uncommon due to high acid and salt content prevent growth of
Clostridia.
 Wild Molds: P. glaucum, P. roquefortii and P. bruneo-violaceum cause
problem if cheese become too dry for P. camemberti to grow; hygienic
precautions essential
Soft cheese
 Soft cheese :Surface Smear-Ripened Soft cheese

 Romadour:
 Made in Austria
 Fat standards 25-45% FDM
 Made in small logs, process similar to Limburger.
 Cheese develop same reddish brown smear by Bre.linens during Short
ripening period (4 wks)
 Proteolytic and lipolytic changes produce ammonia, fatty acids and other
volatile organic compound responsible for odor and flavor.
Soft cheese
 Unripened Soft cheese50-80% moisture)-e.g. Mozzarella, Ricotta etc.
 Made from cream, milk or skim-milk
 Allows a wide range of fat and moisture contents.
 High fat varieties generally have more flavor and softer and smoother.
 Characterized by a high moisture content which contribute to their soft
body and to their perishability.
 Refrigerated storage and transport can prolong their shelf –life.
 Starter bacteria: Str. lactis, Str. thermophilus or Str. faecalis and Lac.
bulgaricus (Mixed Starters)
 Starter bacteria and rennet make a soft from which some whey is drained.
 Salt is mixed into the curd and various flavoring materials added
 Curd is packed in paper cartons or plastics pots and consumed within few
days.
 Some varieties of soft cheese are homogenized to increase the
smoothness of the product.
Soft cheese
 Unripened Soft cheese50-80% moisture)-e.g. Mozzarella, Ricotta etc.
 Microbiological Changes:
 Concerned with the production of acid by suitable lactic streptococci.
 Fast production of acid is required to produce clean, acid flavor, in the
drained curd and starters containing cultures of Str. lactis and Str. cremoris
and aroma forming bacteria like Leuconostoc are suitable.
 Str. lactis sub-sp. Diacetylactis often included in the starter but may
produce yoghurt flavor defect due to acetaldehyde production at certain
times of year.
 Mozzarella cheese: Starter culture of Str. lactis , Str. thermophilus and Lac.
Bulgaricus.
- Rapid acid production is required during manufacturing, eaten fresh.
- Cheese has soft waxy body and mild slightly acid flavor.
 Defects: Post pasteurization contamination , defects in these unripened
soft cheese arise mainly from contamination by yeast and molds.
Miscellaneous
 Cottage cheese
 White brined cheese e.g. Feta, Whey cheese e.g. Ricotta, Processed
cheese e.tc.
 Processed cheese:
 Made from mixed or single variety of natural cheese.
 A cheese mix is prepared to give required properties and flavor, to this
emulsifying salt are added if desired whey powder or skim-milk powder.
 Amount of water is controlled by the final composition of the product.
 Mixture heated to 80-85°C for 4-8 min. filled directly into cans, tubes or
moulds lined with aluminum foil.
 These cheese are packed in suitable outer cases and cooled before going
to store, no ripening required.
 Low moisture and high salt content gives it good keeping quality.
 Only microbial spores survive the high temperature of cooking e.g.
Clostridia- gas production, rancidity and putrefaction result from its
growth
Normal Flora of Cheese Milk
 Bacterial flora of pasteurized cheese milk finally consists of thermoduric
organisms which have survived pasteurization.
 Corynebacteria
• Micrococci
• Enterococci
• Spores of Bacillus and Clostridium
 Other post pasteurization contaminants such as;
• Other Micrococci
• Coagulase-positive Staphylococci
• Coliforms
• Lactic acid bacteria ( Lactobacilli, Pediococci, Leuconostoc and
Enterococci)
 These organisms are derived from the pipe-lines, cheese vats, utensils, air,
washing water and dairy personnel.
 However, cheese is manufactured in large scale in closed vat with
mechanized process the level of contamination is much lower compare to
manual operation.
Normal Flora of Cheese Milk
 Level of hygiene plays in the creamery plays important role influencing the
level of post-pasteurization contamination.
 There is also a necessity to control phage proliferation, esp. in large
factories where large vats are re-used at least once a day; strict hygiene
must be maintained.
 In addition to mos, heat resistant proteolytic and lipolytic enzymes will be
present .
 These enzymes are derived from psychrotrophs which are themselves
killed by pasteurization, survive and remain active in the milk.
 Thus, all organisms present in the milk becomes a part of fresh curd flora,
being concentrated about 10 times in curd.
 Level of these non starter bacteria present in the curd is related to history
of raw milk (e.g. cooling, refrigeration, age etc.), heat treatment it
receives and level of hygiene.
Role of starter culture in
cheese manufacturing
 Production of cheese depends on fermentation of lactose by lactic acid
bacteria (LAB) to form mainly lactic acid.
 This process imparts
 Flavor to cheese curd,
 Assist in the formation of rennet coagulum by causing shrinkage of the
curd,
 Moisture expulsion and
 Promotes characteristics texture formation during cheese making.
 Low pH of fresh cheese curd (5-5.2) helps to suppress the growth of
pathogenic and spoilage bacteria and preserves the products.
 Lactic acid bacteria also imparts
 Traces of flavorful aromatic compounds
 Their proteolytic activity and lipolytic activity (lesser) aids in the
maturation of cheese.
Role of starter culture in
cheese manufacturing
 Lactic acid bacteria also produces
 Low oxidation-reduction potential (Eh) , necessary for the production of
sulphur compounds like methanethiol; contribute to aroma of Cheddar
cheese..
 In old days the fermentation of lactose in milk for cheese making
depended on natural contamination of the milk with LAB from
environment.
 In the modern time, where huge amount of milk is processed into cheese a
rapid and reliable fermentation is needed.
 Since Pasteurization kills the natural LAB hence it is replaced by LAB
deliberately inoculated into cheese milk.
 Selected strains, with predictable acid development and production of
flavorful products are used as starters to obtain a steady rate of acidity
throughout the curd making process.
Types of Starters
 Lactic acid bacteria are used as starter in cheese making including;
Streptococci, Leuconostoc and Lactobacilli.
 Selected strains of these genera are used as
 Combined cultures or
 Single strains cultures or
 Mixtures of single strain culture .
 Mesophilic starter (optimum temperature 20-30°C) are used to
produce a wide variety of cheese.
 For Hard cheese, selected strains of Str. cremoris gives cheese of
good flavor and are free from off-flavors, but if paired with strains
of Str. lactis can shorten the manufacturing time.
 Streptococcus lactis sub-sp. diacetylactis or Leuconostoc which
produce CO2 gives desired open texture for mold ripened cheese
or slight eye formation for Dutch cheese with the production of
diacetyl a flavorful compound.
Types of Starters
 Thermophilic starters (optimum temperature 37-45°C) are used
for production of cooked cheese varieties (e.g. Swiss and
Parmesan), where the starter must withstand a high cooking
temperature of ≥45°C and grow at relatively high
temperature.
 It is quite essential to have a satisfactory starter which
 Produces sufficient acid at a rate required for particular
making process,
 Doesn’t produce off-flavors or bitterness
 Provide conditions in the curd which are suitable for typical
flavor development.
 Starters multiply during cheese making from about 10^7
cfu/ml in milk to 10^8-10^9 cfu/g of curd; growth checked by
salting stage.
Types of Starters
 All strains of starter bacteria are inhibited by antibiotics for e.g.

Penicillin, Aureomycin etc. which may be present in milk following
the treatment of cows for mastitis.
 Penicillin is the most powerful drug and 0.05-0.10 unit/ml milk
affects the growth of starter bacteria.
 These antibiotics can be detected by assay tests.
 Many strains of starters are inhibited by agglutinins and
peroxidases, naturally occurring in some milk.
 Such strains grow better in pasteurized milk as the inhibitory
substances are inactivated by the heat treatment.
Lactic acid bacteria Employed as Starter
Cultures
Bacteria Examples of Usage
1.Mesophilic Starters†
Streptococcus cremoris, Streptococcus lactis Hard pressed cheese, e.g. Cheddar, Gouda.
or Mold-ripened e.g. Stilton
Str. lactis Soft ripened e.g. Camembert, Feta, many
Str. lactis sub-sp. diacetylactis others
Leuconostoc spp. Soft, Unripened, e.g. Coulommier, cottage
Str. cremoris cheese, Quarg, Cream cheese
Str. lactis sub-sp. diacetylactis
L. cremoris
2.Thermophilic Starters†† Swiss-type cheese, e.g. Emmenthal; Italian,
Streptococcus thermophilus with very hard e.g. Parmesan;
Lactobacillus helveticus, Lactobacillus lactis Semi-soft, smear types, e.g.
or Lactobacillus bulgaricus Limburger
3. Mixed Starters Italian pasta filata type e.g. Mozzarella,

Str. Lactis, Str. thermophilus or Str. faecalis Provolone
and Lac. bulgaricus
† Optimum temperature 20-30°C
†† Optimum temperature 37-35°C
Ripening of Cheese
 Ripening involves changes in chemical and physical

properties of the cheese along with characteristics flavor.
 Fresh, young or “green” cheese curd is tough and rubbery.
 Consists of protein, fat and moisture in varying proportions
depending on cheese, along with small amount salt, lactose,
lactic acid, whey protein and minerals.
 During ripening; curd is gradually digested by enzymes and
the mature cheese acquires firm or plastic or soft body
texture of particular variety.
Ripening of Cheese
 Major chemical changes responsible for ripening are:
1. Fermentation of lactose to lactic acid, small amounts of acetic and
propionic acid, CO2 and diacetyl
2. Proteolysis and
3. Lipolysis
 These changes are brought about by enzymes from
I. Lactic acid bacteria of the starter culture
II. Miscellaneous, non-starter bacteria in milk
III. The rennet, rennet paste or rennet substitute used in the milk
IV. Milk itself and
V. Other micro-organisms growing within or on the surface of the cheese.
 These metabolic changes are accompanied by the development of
characteristic flavor
 Affected by the size and composition of the young cheese, and are
controlled by the conditions of temperature and humidity at which the
cheese ripened and stored.
Ripening of Cheese
 For e.g.
 Block stacking of warm cheese on pallets and block stacking of pallets can
influence temperature and flavor differences between blocks of cheese
from the same making vat.
 Some varieties like Emmenthal, Camembert and Stilton requires special
periods of controlled temperature and humidity for ripening process,
during which bacterial and fungal activity produces specific changes in
the body, texture and flavor of cheese.
 Ripening followed by storage, for the hard cheese like Cheddar and
Parmesan constant storage temperature is required throughout the
ripening period and maturation may extend over many months.
Ripening of Cheese
 Major changes during Ripening of Cheese
A. Changes in Body, texture and Flavor
 Body: means consistency of cheese, includes attributes such as firmness,
elasticity, plasticity and cohesiveness.
-Transformation of tough, rubbery curd is brought about by enzymic
digestion of the casein, cheese becomes softer and if moisture is high
crumbly texture is achieved.
- Large amounts of acid developed during manufacturing produce crumbly,
brittle curd and body of such cheese is described as “short” (lacks
elasticity).
- This is defect in some types of cheese for e.g. Cheddar and Emmenthal
where acid development is carefully controlled while it is desirable in
Stilton and Roquefort.
Ripening of Cheese
 Texture : Describes the structure or presence of holes within the cheese.
- Close texture refers to cheese with no holes while cheese with holes is
said to have open texture.
- Where this occurs from failure of curd particles to fuse together, it is
described as mechanical openness and if more than slight it is defect.
- For e.g. In Blue-veined cheese an open texture is necessary to allow
growth of molds throughout cheese.
- If open texture occurs from unacceptable gas production it is a defect.
- However, moderate openness due to gas production is character of some
cheese like Limburger and Gouda and essential for Emmenthal and
Gruyere.
Ripening of Cheese
 Flavor: Ripening of cheese involves development of desired flavor, aroma
compounds by the action of mos and enzymes which break down protein,
fats and carbohydrates and in some cases, metabolize lactic acid, lactate
and citrate.
- Various products of protein hydrolysis as well as fatty acids and their
esters or ketones may be present in varying amounts in the cheese.
- This produces a complex mixture of component which give the required
balance of flavor, characteristic for the variety.
- Starter bacteria dies out during ripening as do most other organisms
present in the curd, like Enterococci and Leuconostoc.
- Only lactobacilli may be present in fresh curd in small numbers, multiply
and these may be present in fresh curd in small numbers, multiply and
reach levels of 10^6 to 10^8/g in cheese in 3-6 wks,
Ripening of Cheese
B. Chemical and Bio chemical changes:
 Most lactose disappears from hard cheese (30-40% moisture) within
first few days of manufacture, but this time may be longer in soft, high
moisture cheese (50%).
 Fermentation of lactose by starter bacteria produces mainly lactic acid,
with some volatile acids, ethanol and small amounts of other by-
products.
 Some lactic acid combines with basic radicals in the cheese to form
salts and in some varieties like Emmenthal where starter includes
propionic acid bacteria, secondary fermentation of lactic acid with
production of propionic acid , acetic acid and carbon dioxide occurs.
Ripening of Cheese
 Most part of nitrogenous material in the young cheese is present as
water-insoluble protein but as ripening proceeds, part or all of this is
hydrolyzed by enzyme action to more simple, soluble compounds
Protein Proteoses Peptones Peptides Amino acids
(insoluble) (---------------------Soluble-------------------------)
 Mos may reduce amino acids to ammonia and organic acids or oxidize
them to form CO2 and amines.
 Extent of proteolysis and formation of resulting compounds helps to
establish the character of mature cheese.
 Soft cheese like Camembert, Brie and Limburger undergoes extensive
proteolysis and formation of water soluble compounds like peptides,
amino acids and ammonia along with high moisture content of these
varieties is responsible for soft velvety texture.
Ripening of Cheese
 Hard cheese like Cheddar and Emmenthal undergo much less proteolysis
and only some 25-35% of protein is made soluble; in a well matured cheese
high proportion of the breakdown products is in the form of peptides and
amino acids.
 Renin can breakdown casein to water-soluble compounds (Proteoses and
Peptones) but microbial enzymes bring about further breakdown with
formation of amino acids and ammonia.
 In Hard cheese mos present are mainly lactic acid forming cocci and rods
which are dispersed throughout the whole of the cheese mass, these
produce extracellular proteinases.
 When these cells die out and autolyse intracellular enzymes are released,
ripening progresses evenly through whole cheese.
 In soft quick ripening cheese most proteolysis is done by extracellular
proteinases released by mos growing in the surface.
Ripening of Cheese
 Molds plays important role in the proteolysis of some cheese (e.g. Blue-veined
cheese Stilton) and surface mold-ripened cheese (e.g. Brie and Camembert).
 Fat decomposition is not extensive, but some hydrolysis do occur during
ripening, important products are volatile lower fatty acids (butyric, caporic,
caprillic).
 Lipolytic enzymes in cheese may have come from milk , mos or from enzymes
preparations added in the milk (e.g. coagulant of bacterial and fungal origin)
 Milk lipase active only in cheese made from raw milk.
 Strains of lactobacilli and other bacteria liberate, upon autolysis, intracellular
lipases, which accounts for most lipolytic activity in hard cheese (Cheddar,
Emmenthal).
 Molds growing in or on the surface of cheese (Stilton or Camembert) are
sources of lipolytic enzymes .
 Rennet extract has little lipolytic activity but rennet paste are actively lipolytic
, hence used in manufacture of Parmesan cheese.
Defects in Cheese
 Spoilage in cheese can be mechanical or biological.

 Types of spoilage can be divided into those
 developed during the manufacture
 ripening of cheese curd and those
 occurring in the finished products
Defects in Cheese
 Spoilage during Manufacture
 During manufacturing of most types of cheese or during draining, a
lactic acid fermentation is encouraged.
 If the lactics are ineffective or contamination with other mos is
heavy abnormal changes may take place which affect the quality of
cheese.
 In Cheese made from raw milk, the gas forming organisms may
produce off flavors as well as gas holes in the curd.
 Lactose fermenting yeast (not present commonly) can also cause
gassiness.
 Spore forming gas producers like Clostridium can cause trouble
either in raw milk or pasteurized milk cheese, if lactic starter
bacteria are not functioning properly.
 Less common : Aerobic spore forming species of Bacillus like B.
polymyxa may produce gas and other defects.
Defects in Cheese
 Spoilage during Manufacture
 Other bacteria may compete with the starter organisms which
do not become evident until curing process, where body and
flavor may be affected.
 Acid Proteolytic bacteria may produce bitter flavor or
Leuconostoc spp may cause openness in Cheddar cheese from
pasteurized milk.
 Cottage cheese is subject to spoilage during storage before
consumption if the starter bacteria produce insufficient acid or
yield it too slowly, cheese becomes inferior due to growth of
undesirable organisms.
 Proteolysis, Sliminess, Off-flavors may ruin the product.
Defects in Cheese
 Spoilage during Ripening
 During ripening cheese undergoes physical and chemical changes
via action of enzyme released from the autolyzed cells of bacteria
that grew during manufacturing and starter.
 Growth of organisms other than desired cause inferior or worthless
cheese due to alteration in body, texture, appearance or flavor.
 Gas Formation: Most common defect is “late gas” caused by
heterofermentative lactic acid bacteria or lactate-fermenting
Clostridium spp.
 Also Bacilli, Propionic bacteria, or heterofermentative lactics.
 Gas holes undesirable in most cheese except Swiss cheese.
 Cracking or splitting of the cheese by gas or the production of too
many, too small or mis-shapen eyes.
 Gas formation by the spore formers is accompanied by the
production of undesirable flavor e.g. butyric acid from the
anaerobes
Defects in Cheese
 Bitterness: Caused by certain lactic streptococci, proteolytic bacteria
e.g. the acid proteolytic types; Coliforms, Micrococci and rarely yeast,
which gives sweet, fruity or yeasty flavor.
 Putrefaction: May occur locally or generally in cheese where
insufficient acidity has been produced by the lactics or the acid has
been destroyed by lactate fermenter e.g. Clostridium tryobutyricum, C.
sporogenes or C. lentoputrescens ( anaerobes)
Defects in Cheese
 Discoloration: Results from the action of mos or compounds produced
during curing or on added coloring materials like annatto in Cheddar
cheese or may result from the development of pigmented colonies or
organisms on or in the cheese.
- Blue , green or black discoloration may be produced by the reaction of
H2S produced by organisms with metals or metallic salts.
- Bacterially formed sulfhydryl groups give a pink to muddy appearance
to annatto.
- Reddish brown to grayish brown color results from the oxidation of
tyrosine by bacteria growing in soft cheese.
- Rusty spots of Cheddar cheese and others are caused by colonies of
Lactobacillus plantarum var. rudensis or l. brevis var. rudensis
- Yellow, pink or brown spots in Swiss cheese mostly on surface of the
eyes are colonies of pigmented sps of Propionibacterium.
Defects in Cheese
 Spoilage of Finished Cheese
 Perishability of cheese increases with the moisture content.
 Soft cheese like Limburger and Brie are most perishable and hard cheese like
Cheddar and Swiss cheese the most stable.
 Most common organisms being molds that tend to grow on the cheese
surface and into their crack holes.
 Even cheese which depend partly on a specific mold for ripening may be
damaged by other molds.
 Most cheese have rinds that serve as some protection to the anaerobic
interior but usually are not dry enough to prevent the mold growth.
 Acidity of cheese is not deterrent to growth and storage temperature is not
too low for such growth.
 Most molds grow in colored colonies on the surface or in the crevices,
without much penetration into the cheese but some produce rots
 Products produced by the mold like mycotoxins and antibiotics can migrate
in to the cheese.
Defects in Cheese
 Molds growing on the cheese surface are:
 Oospora (Geotrichum spp.) (Geotrichum lactis): Dairy mold, grows on
soft cheese and during ripening sometimes suppresses other molds as
well as surface-ripening bacteria. The curd gradually becomes liquefied
under felt O. rubrum, O. crustacea produce red coloration and O.
aurianticum forms orange to red spots.
 Cladosporium spp.: The Mycelium and spores of these molds are dark or
smoky and give dark colors to the cheese. E.g. C. herbarum
characterized by the dark-green to black colors. Others cause green,
brown or black discoloration.
 Penicillium spp.: P. puberulum and other green spored species grow in
cracks, crevices and holes of Cheddar cheese and other related cheese
to give a green coloration because of their spores. May act on annatto
to cause mottling and discoloration. P.casei causes yellow-brown spots
on the rind and P.aurantio-virens discolors Camembert cheese.
Defects in Cheese
 Molds growing on the cheese surface are:
 Monilla spp.: M. niger produces penetrating black spots on the rind of
the hard cheese. Species of many other genera may discolor cheese
and give off-flavors. E.g. Scopulariopsis, Aspergillus, Mucor and
Alternaria.
 If the surface is sufficiently moist yeast may form colored colonies or
areas and film yeast may pave the way for the yellow to red growth of
Brevibacterium linens in surfaced ripened cheese.
Bacteriophage Contamination in
Cheese
 Bacteriophage are virus that infect bacteria.
 Can cause complete lysis of bacterial culture.
 Obligate intracellular parasites that multiply inside bacterial cell
by making use of some or all host biosynthetic machinery.
 Mode of replication: Lytic (or virulent phage) and temperate
phage (Lysogenic phage)
 Most phage multiplication cycle end with cell lysis and the release
of hundreds of new virions ready to infect neighboring cells.
 Problems due to the presence of phages were reported in the
food, chemical, pharmaceutical, feed and pesticide industries.
 The dairy industry is probably the one in which phage problems
are the most documented.
Cheese
 The manufacture of cheese requires the inoculation of
carefully selected bacterial cells (known as starter cultures)
per ml of pasteurized milk to control the fermentation and
to obtain high-quality end-products.
 Starter cultures are a combination of various lactic acid
bacteria (LAB), usually strains of Lactococcus lactis,
Streptococcus thermophilus, Leuconostoc sp., and/or
Lactobacillus sp.
Cheese
 In the non-sterile environment of raw or heat-treated milk,

the added LAB cells will come into contact with virulent
phages found in milk .
 Although phage concentration is usually low in milk, a
specific phage population can increase rapidly if phage-
sensitive cells are present in the starter culture.
 The consequent lysis of a large number of sensitive cells will
delay or even halt the milk fermentation process leading to
low-quality products.
 In worse cases, the inoculated milk must be discarded.
Cheese
 The dairy industry has been dealing with this natural

phenomenon and has relied on an array of control
measures
 notably adapted factory design,
 improved sanitation,
 process changes,
 specific culture medium,
 strain rotation, and
 use of phage-resistant strains.
Cheese
 Sources of phage contamination
 Phages can come from various sources.
 It is of prime importance to know the potential sources of
phages to limit their entry within the manufacturing
facilities, which could be deleterious to the fermentation
process.
I. Raw ingredients
 Any raw natural ingredient that enters a fermentation
facility may contain phages, albeit at low levels.
 For example, raw milk, which is an ecological niche for some
LAB, is well known to contain phages.
 Milk is collected from different farms, phage biodiversity is
amplified within milk silos.
Cheese
I. Raw ingredients
 Phages can easily propagate in a liquid medium such as
milk as they can also diffuse in gel-like media, only a few
sensitive cells are needed to rapidly increase phage levels
in a given environment.
 These numbers can be higher in whey samples or final
products since phages can propagate during most
fermentation processes.
 Titers as high as 10^9 PFU per ml of cheese whey have
been reported.
Cheese
I. Raw ingredients
 Depending on the frequencies of phage attacks and the
size of the facilities, it may be advisable to analyze
-Milk (or other ingredients) for the presence of phages
before beginning the fermentation process to confirm
that the initial phage load does not represent a significant
risk of fermentation failure.
- If the ingredients are thought to pose a risk, they can be
treated to reduce phage levels or used for other processes
that will not be affected by phages.
- Effective cleaning procedures must be also in-place to
reduce to the initial phage load.
Cheese
II. Processed or recycled ingredients
 The milk fermentation industry may reuse whey proteins
to improve the taste or texture of a final product to increase its
nutrient value, to standardize, milk before the fermentation
process or to increase the yield .
 Upon whey or milk protein concentration, phages may remain
in the whey protein concentrate (liquid or dried) and
contaminate the products to which it is added.
 When using membranes to separate whey components, it is
highly possible that phages will be retained by ultra filtration
and/or microfiltration.
Cheese
II. Processed or recycled ingredients
 Ideally, milk by-products should either be treated to inactivate
the phages or be used in a type of fermentation that is driven
by different starter cultures.
 For example, if the whey was collected from a cheddar
fermentation made using mesophilic starter cultures, by-
products of this whey could safely be used in yogurt
manufacture or in a cheese process requiring thermophilic
cultures.
 In addition, the use of concentrated milk products from
another dairy plant (which may use different starter
cultures)can offer additional protection. Although the latter
will also most likely increase the phage biodiversity within the
factory.
Cheese
III. Phage reservoir
 One perceived source of phages is the starter culture itself.
 When a temperate phage enters a strain, it can either start
the lytic cycle or its genome can integrate into the bacterial
chromosome and follow bacterial multiplication.
 When bacteria carry such a prophage, the cell is called a
lysogen.
 Different bacterial stresses such as heat, salts, antimicrobials,
starvation or UV can induce the prophage and trigger the lytic
cycle .
 Thus, the use of lysogenic strains in a starter culture may lead
to cell lysis during fermentation.
Cheese
III. Phage reservoir
 Induction can also occur naturally and can reach a frequency of up to 9% .
 Prophages are carried by many LAB strains and often more than one
prophage is found in a genome.
 The most recent analysis revealed that 25 out of 30 commercial,
collection or dairy isolated Lactobacillus casei, Lactobacillus paracasei and
Lactobacillus rhamnosus were found to carry inducible prophages .
 It should be noted that most starter culture suppliers will test their
strains for the presence of prophages and their natural induction rate.
 Usually, lysogenic strains carrying easily inducible prophages will not find
their way into commercial products.
 Of note, phage induction assays cannot be readily performed with
undefined starter culture as the exact strain composition of this type of
starter is unknown.
Cheese
IV. Air/surfaces
 Dissemination routes of contaminants can be more complicated to identify.
 The presence of airborne lactococcal phages in a cheese plant was
investigated because it had been rarely documented .
 A high level of airborne phages in the environment may mean that phage
propagation has previously occurred or that phage problems are likely to
occur.
 The large amount of milk processed daily in open cheese vats, as well as
whey processing, inevitably lead to liquid splashes and aerosolization of
phages .
 These bacterial viruses can also be aerosolized by air displacement around
the surfaces of contaminated fluids and transported elsewhere in the plant.
Cheese
IV. Air/surfaces
 No standard procedures have been established to detect airborne viruses,
which can be present in a wide range of particle sizes, from nanometer to
micrometer.
 Hence there is a need for testing sampler and detection methods in a
particular environmental setting as each have their advantages and pitfalls .
 Air samplers were recently tested in a cheese factory and samples were
analyzed for the presence of lactococcal phages using qPCR.
 Air sampling was performed for 12 hours next to the filling section at the
end of a cheese production line
Cheese
Methods of detection
 New protocols are still being designed with improved efficiency for detecting
phages in industrial processes.
 Microbiological methods, such as plaque assays or acidification
monitoring, have long been the gold standard for phage detection because they
are quantitative and sensitive methods, though time consuming.
 Other methods:
 PCR-based methods:
-Classic PCR detection methods were successfully used to detect or to quickly
classify Lactobacillus, Lactococcus and Streptococcus phages.
-These methods could be used directly on milk or on whey samples to detect the
presence of phages.
-The lowest detection limit reported using a classical one-step PCR method is 10^3
PFU/ml but this usually varies from 10^4 to 10^7 PFU/ml depending on the
phages tested and the nature of the sample.
- qPCR-based methods can overcome this inconvenience by monitoring the
replication of specific phage genes in real time, during the fermentation.
Cheese
 Other methods:
 PCR-based methods:
-Classic PCR detection methods were successfully used to detect or to
quickly classify Lactobacillus, Lactococcus and Streptococcus phages.
-These methods could be used directly on milk or on whey samples to
detect the presence of phages.
-The lowest detection limit reported using a classical one-step PCR
method is 10^3 PFU/ml but this usually varies from 10^4 to 10^7
PFU/ml depending on the phages tested and the nature of the sample.
- qPCR-based methods can overcome this inconvenience by monitoring
the replication of specific phage genes in real time, during the
fermentation.
Cheese
 Other methods:
 Impedimetric monitoring
-Promising new biosensor technologies were recently developed to detect
phages (biosensor).
-The technique is based on the binding of phages to bacterial cells attached to
a chip.
 Flow cytometry
-A novel technique using flow cytometry was also designed to incorporate the
host specificity of phages.
-When phages infect their hosts, these bacteria undergo morphological
changes leading to lysis.
-The loss of mass and the interruption of cell division are two changes that can
be monitored by flow cytometry .
Cheese
Flow cytometry
-While low contrast cells can be observed under a phase-contrast microscope
following phage lysis, the light scattering of the flow cytometer can
efficiently measure the mass of the cells, as long as the bacterial chains (in
the case of LAB) are first broken by vigorous shaking.
-This property allows the flow cytometer to discriminate the infected from the
non-infected cells.
-To assess the presence of phages, the culture is run on the flow cytometer,
which gives the distribution of the cells’ mass.
-A broad distribution of cell mass indicates the presence of both lysed and live
cells whereas live cells will typically give a narrow peak.
-This technique has the advantage of detecting the cell morphology changes
regardless of the strain, the phage or the number of strains in a starter
culture.
- The reported detection limit (10^5 PFU/ml) was comparable to classical PCR
Cheese
Control
 Sanitation
 Raw material treatment
 Starter rotation
 Anti-phage mechanisms
 Phage inhibiting components
Microbiology of condensed
Milk
Microbiology of condensed Milk
 Condensed milk is cow's milk from which water has been
removed.
 It is most often found in the form of sweetened condensed
milk, with sugar added, and the two terms 'condensed milk'
and 'sweetened condensed milk' are often used
synonymously today.
 Sweetened condensed milk is a very thick, sweet product
which when canned can last for years without refrigeration
if unopened.
 Though there have been unsweetened condensed milk
products, they spoiled far more easily and are uncommon
nowadays.
 Condensed milk is used in numerous dessert dishes in many
countries, including the United States, India, Germany,
Argentina, Brazil, Venezuela, Viet Nam, China, Lebanon,
and Russia.
 A related product is evaporated milk, which has undergone
a more complex process and which is not sweetened.
 Local tastes in most countries prefer one or the other.
 In Germany unsweetened evaporated milk is far more
common than sweetened condensed milk. In Peru and the
US both are equally common.
Microbiology of condensed
Milk
Objectives:
1. To improve the shelf life and keeping quality of milk
2. To reduce the cost of transportation.
3. To save the storage and package space.
4. To serve as resource material in scarcity conditions.
5. Ready reconstitutability to convert back into milk
6. To serve as an additive for certain products preparation i.e. ice-
cream, candies and food items.
Standard and Specification for
Condensed Milks
 The first and most imp prerequisite of evaporated milk or
condensed milk is that it must be sterile. The temperature
treatment applied in the process must be adequate to ensure
complete destruction of all microbes present in milk.
 Each can must receive full sterilizing effect……This is achieved
by uniform distribution of heat to all cans by uniform
distribution of steam over entire length of sterilize and
absence of air pockets between cans.
 Air pockets are avoided in batch sterilizer by presence of
water ( water hasten up the coming up period shortening the
time by above 5 mts)
 Avoid leakers……Seals and seams must be tight because this is
the major cause for spoilage.
Manufacture of Condensed Milk
1.Raw milk supply
 Initial quality of raw milk has a direct bearing on finished product.
It must have MBRT not less then 2.5 hours RRT and DMC also are
used for assessing the quality. Standards are more lenient than
those for fluid milks, because of opinion that finally product is
sterilized.
 Raw milk should be cooled to <4.40C to avoid development of
undesirable changes. Holding for>24h is discouraged to prevent
growth of Psychrophiles.
 Sanitary quality of milk at the receiving platform depends on its
production background on the farm. Viz: Healthy cows, clean milk
production, clean utensils, freedom from colostrum, prompt
cooling and refrigerated transports.
1.Raw milk supply
 All milk supplies must be systematically and thoroughly inspected
each day by conscientious and experienced milk graders.
 It must be Clean and Sweet It should be free from off-flavors &
odors and free from extraneous material.
 Reject milk with excessive acidity, increase bacterial count or
decrease heat stability.
2. Filtration /clarification:
 Done at 35 to 40◦C. It helps in removing visible foreign matter and
removing somatic cells and some bacteria and has no significant
change in microbiological quality.
3. Standardization
 The materials used to standardize i.e fat: solids ratio, whether
cream, or skim milk should be of the same or of better quality than
the raw milk.
 Proper provision must be made for the production and handling of
these products.
4. Fore warming/ Pre heating
 The milk proteins are stabilized against coagulation by heating to
above 93.40C/20- 25 min. which is done in a ‘hot wells’ or ‘surge
tanks’ or for continuous operations up to 1210C /few min. This
temperature destroys
 All non-sporulating bacteria present
 Many of the less-resistant sporulating types
4. Fore warming/ Pre heating
 Fore warming temperature. apparently constitute a sub lethal heat
shock which makes the spores sensitive to the final heat treatment.
 Bacteria causing infectious disease
 Thermophiles is not a problem – high temperature will not allow the
growth of Thermophilic bacteria
 The enterotoxins produced by entero toxigenic staphylococci would
not be inactivated. So proper precautions necessary to prevent
extensive growth the organism in milk.
5. Condensing / Evaporation:
 Two types: Batch and Continuous
 Temperature during the process seldom exceeds 54.50C and will not
kill those bacteria which survived preheating. Thermophilic bacteria
may show moderate growth in these conditions
5. Condensing / Evaporation:
 Growth of Thermophilic bacteria becomes a factor in limiting the length
of time the equipment can be operated.
 Operation for extended periods may also result in extensive build up of
solids on the heating surfaces that satisfactory cleaning will be made
quite difficult .
 The accumulated soil on equipment surface may offer protection to
undesirably microbes that may develop later.
6. Homogenization:
 Pressure has little effect on micro organisms. Usual precautions
concerning homogenizer care must be observed to avoid excessive
contamination from this source.
 Carelessly cleaned homogenizer may be important source of coliform
and other bacterial contamination. After the use homogenizer is
cleaned with tepid water. All valves, pistons, cylinders, pipes should be
scrubbed with a hot solution of washing powder, rinsed and allowed to
dry.
7. Cooling:
 Cooling and holding under good refrigeration conditions until
packaging and sterilization is essential because the product is not
sterile and can thus spoil if conditions permit appreciable microbial
growth.
8. Pilot Sterilization:
 To determine the amount of chemical stabilizer to add i.e Tri Sodium
Citrate (TSC) and di Sodium Phosphate (DSP)
9.Canning/Packaging:
 Cans if fabricated in adjacent plant, the heat employed in fabrication is
enough to ensure that the cans are free of microbes.
 Sufficient headspace is allowed above the product in cans, because milk is
cold when placed in the can, expansion occurs during sterilization
process, since during expansion the cans may because leaky by opening
one or two seams due to internal pressure and permit the entry of
bacteria when heat level is no longer adequate for destruction.
 Hermetic seal is essential i.e. to avoid air or fluid in either direction or a
non-hermetic seal is an invitation to contamination.
 Latest technology is to sterilize the milk which is followed by aseptic
packaging of milk. The fillers used for placing the milk in the final
container are complex and difficult to clean properly.
 Recommended procedure is complete disassembly of the canner and
careful cleaning of each of the component part. In the absence of sound
practice filler can contribute enough bacteria of wrong types.
 So increases in severity of heat treatment to get “commercial
sterilization” of the final product.
10. Sterilization:
 To have adequate keeping quality at room temperature, evaporated
milk must be “commercially sterile”. That means it must not contain
organisms which will grow and probably produce defects in normal
storage conditions.
 Time-Temperature combination varies to produce ‘practical sterility’
or ‘commercial sterility’.
 “Absolute sterility would be associated with an unacceptable level of
“cooked “flavor.
 A dark color and some modified physical characteristics. So the
temperature-time chosen is the minimum one that will provide
commercial sterility and keep changes in flavor and physical
characteristics to a minimum. when spore load of milk is high, the
exposure must be greater than normal.
 Presence of unusually resistant spore also increases the exposure.
The temperature generally followed are 115.5 oC for 20 min ( 114.5-
118.4 oC for 14 -18 min).
10. Sterilization:
 Gales loot (1962) observed that by increasing temperature from 120 to 150oc
there was an increase in spore destruction rate by 100 fold and browning rate
by 15.7 fold.
 Markedly reduced time of exposure required at higher temperature gives
equivalent bactericidal effect retention of flavor and color.
 Disadvantages of higher temperature: Reduces viscosity – sometimes some
fat separation; Reduces shelf stability – so gelation may occur.
 To over come the problems of low viscosity and gelation the historical
procedure for sterilization has been to hold the canned product for 15-20 mts
at 1150C or slightly higher.
 Recent Technology is the use of UHT treatment
-Temperature: 130◦C /30 sec or 150◦C / ‹1 sec
-It must kill ‘normal’ load of Mesophilic spore forming bacteria.
-The most resistant in this group is “Bacillus subtilis”. But highly heat resistant,
obligately Thermophilic spore former i.e “Bacillus stearothermophilus” survive in
small number but do not grow and cause defects under normal holding
conditions for canned evaporated milks.
 Aseptic packaging of UHT products poses numerous microbiological
problems:
 The container & closure must be sterile.
 The equipment through which the product passes must be sterile.
 Contamination by micro organism from air must be avoided.
 Prevent entry of air & water and contamination from other sources by
sealing container hermetically.
 Avoid rough handling subsequent to filling to prevent damage to the
package or even temporary weakening of the closure which may permit
contamination.
 Metal cans - may be flame heated or autoclaved with super heat to be made
sterile.
 Glass containers – cleaned thoroughly and then autoclaved.
 Paper – plastic – foil containers – treated with H2O2 and dry heat. For dry
heat air heated to 200o C effectively remove any residual peroxide.
 An atmosphere of superheated steam or hot gas must be maintained
around the filler, closing machine and interconnecting conveyor system to
preserve sterility.
Microbiological Examination
 Customary testing procedures after packaging show no viable organisms.
 Microorganism seldom develops even after prolonged holding at usual room
temperature, although defects do appear occasionally.
 In past years – before shipping – to detect spoilage canned condensed milk
was held for 2-3 weeks.
 Improved technology – Representative samples are held for incubation and
examination. Incubation – 37 or 55oc are used to detect spoilage organisms
like facultative or obligate Thermophilic bacteria which survived heat
treatment.
-The contents of pack are removed aseptically and small quantity smeared onto
plates into already poured agar plates. – incubated at temperature which the
packages had been held. Usually no quantitative method is followed.
 The presence of viable organism is a criterion. Presence of Non-spore
forming organisms indicates post-sterilization contamination due to leaky
containers and Spore formers indicate inadequate heat treatment.
 The packs are examined for organisms survived heat treatment and
organisms as contaminant of aseptic packaging and organisms gained entry
as result of leaky container.
Type of flora in condensed milks:
 Among the variety of micro organism in the milk samples, the most
predominant types are invariably the heat resistant bacteria.
Contaminating organisms especially non-spore forming types may
also gain access into the milk due to defective cans.
1. Heat resistant bacteria:
B. cereus, B. subtilis, B. coagulans, B. circulans, B. licheniformis, B. brevis,
B. polymyxa, B. mesentericus, B. mycoides, B. stearothermophilus B.
amarus, B. panis , Cl. sporogenes Cl. butyricum Cl. foetidum Cl. Botulinum
2. Post processing contaminants: E.coli (gassy type), Ps. Ichthyosmia,
Enterobacter sp., St. distendens, Certain yeasts.
MICROBIAL DEFECTS
 Microbial defects are due to
 Heat resistant organisms that survive slightly inadequate heat
treatment
 Organisms which gain entrance after heat treatment.
 Inadequate heat treatment may be due to:
A. Insufficiently intense time time-temperature combination
B. Deviation from chosen sterilization programme. Eg: temporary
decrease in sterilization temperature during holding period.
C. Due to uneven distribution of heat in sterilizer. So Thermophilic and
heat resistant spore forming bacteria may survive and serve as spoilage
producers.
 Contamination is mainly due to leakage of cans as result of improper
seal, damage to the tin.
MICROBIAL DEFECTS
1. Gassy fermentation or Bloat
 It is the most serious defect of evaporated milk.
 Signs: Bulging of cans.
 Organism: Anaerobic spore forming gas producers. Eg: Cl. foetidum.
This organism also causes putrefactive changes along with gas
production (butyric acid and putrefactive types.) It produces hydrogen
per sulphide which gives foul smell.
 Bulged cans are also much more commonly by a chemical action on the
metal of can, or by overfilling of the cans with cold milk which then
expands on heating.
 Other bacteria are: B. coagulans, Streptococcus sp.
 Prevention:-
1. Improved sanitation in production of milk on farm as manure, soil, crop
residues are important source of anaerobic spores.
2. Proper time temperature combination for sterilization.
MICROBIAL DEFECTS
2. Bacterial coagulation:
 Sweet or sour type.
a) Sweet coagulation:- sweet curdling spore formers Eg: B. subtlis.
 It causes non acid curd, which may then be digested to a brownish
liquid with a bitter taste by proteolytic enzymes.
b) Sour coagulation:- due to acid producing spore formers
The changes are
 coagulation due to microbial acidity
 sour and cheesy taste
 flat sour bacteria which sour milk but form no gas and no bulging.
MICROBIAL DEFECTS
 B. cereus – slightly soft curd at the surface of the milk in can. It is highly
aerobic. More the head space the more the spoilage.
 B. coagulans – causes increase in acidity as it grows at 37oc. Produces
acid coagulation and slight cheesy odour and flavour
 B. megaterium – coagulum formed is accompanied by some gas and a
cheesy odour.
 B.subtlis : non acid coagulation.
 B.stearothermophilus – grows at thermophilic temperature. Acidity and
cheesy odour is produced.
Microbial Defects
 Predisposing factors:
o High temperature of storage
o inadequate heat treatment
o improper cooling
o contaminate with heat
o Resistant organism.
 Prevention
o cold temperature storage
o proper sterilization
o adequate cooling
Microbial Defects
4. Bitterness:
 Facultative heat resistant spore forming bacteria survive
marginal heat sterilization and grow satisfactorily at room
temperature and causes break down of protein and formation
of peptones and other decomposition products and is
sometimes accompanied by wheying off.
Organisms:
 B. subtlis - Bitterness and thinning
 B. amarus - Bitterness and abnormal flavour.
 B. panis - Bitterness.
 Prevention:
o Proper sterilization
o Proper heat distribution.
 Post processing contamination defects:
 Breakdown of aseptic canning process by Spore former and Non spore
former.
 Leaky cans contribute to non spore formers due to improper closing of
hermetic seal, Subsequent corrosion, Mechanical injury, Momentary leak.
Precaution:
o Once product is opened it should be refrigerated.
o Maintain pressure while cooling to avoid bulging to prevent weakening of
seals to prevent entry of cold water into cans.
 Non-microbial defects:
 These are cooked flavour, coagulation, browning, discoloration, age
thinning, mineral deposits and bloats.
 Public health significance: Most important concern is of Cl.botulinum
 Anaerobic conditions in the can are congenial for the growth and
proliferation of this organism.
Quality control tests: SPC, Anaerobic spore count, Sterility test.
SWEETENED CONDENSED MILK (SCM)
 SWEETENED CONDENSED MILK (SCM) : Made from whole milk
and from skim milk.
 Water is removed by evaporation and sugar is added to yield a
product with high solute concentration to prevent growth of most
microorganisms.
 Codex Description: Sweetened condensed milks are
 Milk products which can be obtained by the partial removal of
water from milk with the addition of sugar, or
 By any other process which leads to a product of the same
composition and characteristics.
 The fat and/or protein content of the milk may have been
adjusted, only to comply with the compositional requirements in
of this Standard,
 By the addition and/or withdrawal of milk constituents in such a
way as not to alter the whey protein to casein ratio of the milk
being adjusted.
 Principle of preservation:
 Addition of sugar increases the osmotic pressure to a point
inhibitory to most microorganisms. The added carbohydrate also
binds water, making it unavailable for metabolic process. This is
explained by the concept of water activity.
 The increased concentration of milk solids also is effective in raising
osmotic pressure and in binding water but this is relatively a minor
compared to the effect of added sugar.
 Absence of air in hermetically sealed containers also contribute to
the keeping quality of the canned product inhibiting the growth of
aerobic microorganisms, particularly molds, a few yeasts and some
micrococci which can tolerate the high osmotic pressure.
 Concentration of sugar in water of SCM is known as ‘sugar ratio’
Ratio usually followed is 62.5 to 64.5 for canned goods
Sugar ratio= % of water in SCM X 100/100 – Total milk solids in
SCM
Manufacture of Sweetened
Condensed Milk
 CODEX STANDARD FOR SWEETENED CONDENSED MILKS
 ESSENTIAL COMPOSITION and QUALITY FACTORS
 Raw materials: Milk and milk powder, cream and cream
powders, milk fat products.
 Permitted ingredients
• Potable water
• Sugar
• Sodium chloride
• Sugar is generally considered to be sucrose, but a combination of
sucrose with other sugars, consistent with Good Manufacturing
Practice, may be used.
(Codex)
 Composition
• Sweetened condensed milk
-Minimum milk fat: 8%
-Minimum milk solids: 28%
-Minimum milk protein in milk solids-not-fat: 34%
 DFTQC Standard
- Minimum milk fat 8%
- Minimum milk fat with solids: 31%
- Minimum sugar content : 40%
US Standard (FDA) for SCM
Physical requirements
(a) Flavor. Shall be sweet, clean, and free from rancid, oxidized, scorched, fermented,
stale or other objectionable tastes and odors.
(b) Color. Shall be white to light cream.
(c) Texture. Shall be smooth and uniform, free from lumps or coarse graininess.
There shall not be sufficient settling of the lactose to cause a deposit on the bottom
of the container.
(d) Body. Shall be sufficiently viscous so that the product upon being poured at
room temperature piles up above the surface of that previously poured, but does not
retain a definite form.
Microbiological limits.
(1) Coliforms, less than 10 per gram;
(2) yeasts, less than 5 per gram;
(3) molds, less than 5 per gram;
(4) total plate count, less than 1,000 per gram.
BIS Standard for SCM
Manufacturing of Sweetened
Condensed Milk
1.Raw milk:
The important criteria for raw milk selection are :
 MBRT not less than 3.5
 DMC Not more than 10^6 cfu / ml
 No physical abnormalities such as blood, flakes, clots
 No developed acidity (not more than 0.03% LA over natural acidity)
 Sediment not more than 0.03 mg
 The addition of sugar should not be considered as a substitute for
good sanitation, good quality milk or adequate processing practices
since sterilization process are not used.
Condensed Milk
2.Fore warming:
 With the increase in concentration the temperature of heat treatment
also vary. Total solids concentration varies between 12 and 25.
Temperature used varies between 82-100°C / 10-30mts. The temperature
and time used are sufficient to destroy all pathogenic microbes and all
other except most heat resistant ones. The z value should be nearly
’10.5’.
 Inactivates the natural enzymes of milk.
 Necessary for satisfactory vacuum pan operation.
 It is only means of destroying pathogenic organisms and is instrumental
in destroying spoilage organisms.
 The bacterial enzymes lipases, proteinases are not inactivated.
 Holding in HOTWELL: 90-105°C / 20 min. No drastic change in SPC and
the survivors could be spore formers, and micrococci
3. Addition of sugar: Condensed Milk
 Addition of sugar to the fore warmed milk before condensing. This
results in plasmolysis of some surviving microorganisms.
 Addition before fore warming – offers protection for destruction of
microorganisms.
 Added to syrup (65% solution) – drawn into the pan towards the end of
run.
 Sugar is sucrose, stored in dry place, free from dust insect and rodent
contamination. Under unfavorable conditions, get contaminated with
mold spores, osmophilic yeasts and microbes that will produce acid and
gas.
Condensed Milk
4. Superheating:
 The temperature is raised to 70°C and held for varying time to obtain
desired viscosity of product. There is little effect on microbial quality
5. Condensing:
 It is done in vacuum pan. Milk boils at 57.2°C at 25 inch vacuum, but
towards end temperature drops to 48.9°C in 3-4hr. Condensing is not
expected to reduce the microbial population significantly.
 Vacuum pan should be cleaned very well. A possible source of
contamination may be vacuum pan heads. Milk deposits (sticky nature)
that quite difficult to clean may become source of contamination.
 Recommended – Alkaline detergent 82°C at 14 inch vacuum followed by
acid detergent and 600ppm chlorine solution
 Finishing – slightly over condensing and adjust with sterile water or
under condensed milk such a process is prone to post processing
contamination.
Condensed Milk
6. Forced crystallization:
 Temperature is 30°C. Milk is seeded with lactose crystals and
vigorously agitated for 1hr, to force lactose to form fine crystals.
 Added lactose is usually not heavily contaminated and SPC should
not exceed 10 to 15/g but can be sterilized by heating under vacuum
at 93.8°C to convert to α –anhydride form, grounded, canned and
sterilized at 130°C/1-2hr.
7. Packaging:
 Filling operations should take place in a separate sanitary room in an
atmosphere of filled air at around 16°C.
 Dust and insect contamination of stored cans and ends must be
avoided.
 Filling operation may be major source of micrococci, Yeasts and
Molds.
Microbiological examination of SCM
 SCM is not a completely sterile. Total count varies between few hundred
to 1,00,000/gm.
 Microbes that can be observed include: Micrococci, Yeast and molds,
spore forming aerobes, coliforms which are due to contamination from
air and utensils.
 Micrococci, spore formers ex: - B. subtilis may survive fore warming
process.
 Sampling of sealed cans: Thorough cleaning of cans, Testing the open
area with microbiocidal agent with alcohol, Use of sterilize opener, Care
in sampling of material prewarming carefully to 45°C to reduce viscosity.
 Three tests are done usually:- SPC, yeasts and molds, coliforms.
 First dilution is made gravimetrically (11gm/99ml) and then volumetric
dilutions (because the product is very viscous) other quality control test
are thermoduric count, tests for micrococci and staphylococci.
Type of Flora:
 Bacteria
1. Micrococci: - Due to marked resistance to unfavorable conditions like
high osmotic pressure prevalent in the product. M.varians, M .candidus,
M. caseolyticus (Predominate) M. luteus M. freudenreichii
2. Staphyococci: - S. aureus, S. epidermidis
3. Coliforms: - present in very few numbers [3%] E. coli, Ent. aerogenes.
4. Aerobic spore formers: - Due to resistance to fore warming
B. subtitis (Predominate), B. cereus, B. mesentericus, B.stearothermophilus, B.
mycoides, B. pseudoanthracis
5.Anaerobic spore formers: - Cl. welchii, Cl. butyricum, Cl. sporagenes, Cl.
thermosacharoiyticum
6. Streptococci: - Enterococcus faecalis, Streptococcus faecium, E. faecalis var
zymogenes
Type of Flora:
7. Others: - Pseudomonas sp, Serratia marcescens

 Yeasts and molds: -
• Yeasts are Saccharomyces, Candida, Torulopsis, Trichosporon,
Rhodotorula
• Molds are
 Asperigillus , Penicillium (Predominates) Cladosporium,
 Catenularia , Mucor
MICROBIAL DEFECTS of SCM
 Fresh SCM SPC varies between 5,000-50,000/g and the reasons for this are
1. Heat treatment used not sufficient to kill spores
2. Organisms due to contamination
3. Enough oxygen may be present in headspace of incompletely filled containers to
support growth of high osmotolerant organisms.
 Common defects are
 Gassy fermentation,
 Bacterial thickening,
 Mold button formation
 Other Defects
 Lipolysis – due to lipases (Fat glycerol +FA); butyric, caproic and caprilic acids
impart objectionable bitter flavor and pungent sharp, unpleasant rancid aroma.
 Yeasts; - Mold – Pencillium sp. Geotrichum sp
 Bacteria; - Ps.fluorescens, lipolytic micrococci, bacillus sp.
 Glue like odor and taste -Thermobacterium mathiacelle
 Fruity flavor
 Fishy
 BUTTON FORMATION:
 Causative organisms: - important species are Asperigillus sp and
Penicillium sp., A.repens, A.glaucus,Cladosporium sp., Catenularia
fulginea, certain actinomycetes
 Buttons are composed of old mycelia and coagulated casein due to the
production of milk clotting enzymes (affecting casein stability) and are
colored white to brown.
 Mold buttons are lumps of variable size, cheesy consistency and
whitish yellow to reddish brown colour.
 They constitute firm self-contained units that do not re emulsify into
the body of milk.
 Buttons are found on surface or subsurface of the product and molds
grow until the availability of oxygen in headspace is exhausted.
 The button formation may continue even after cessation of mold
growth, presumably due to continued enzymatic action.
BUTTON FORMATION:
 Signs: -
 Lumps or buttons of dark brownish colour,
 Disagreeable taste, stale odour accompanied by colour change, flavor
change and
 Defects in body and texture.
 Major source: -
 Post processing contamination, because almost all molds are destroyed
by fore warming operation
 Storage at high temperature
 Presence of air in headspace of cans
 Prevention: -
 Filling cans fully
 Vacuum packing
 Storing below 16°C
 Improving sanitation of plant
 Prevention of contamination after fore warming
 Inversion of stored cans at regular intervals.
 GASSINESS:
 Gas may develop suddenly in the product after 10 days to few weeks.
 Concentration of the most disastrous defect
Signs: -
 Gas cause bulging of cans, blowing of cans.
 In extreme cases may break the cans or burst the barrels,
 Extreme acidity, lumpiness and
 Darkening of internal surface of containers
Organisms: -
 Yeasts -- Torulopsis lactis condensi, T. globosa that ferment sucrose but
not lactose
 Bacteria – coliforms, B. cereus, B. butyricus, B. coagulans, Cl. butyricum
GASSINESS:
Mechanisms: -
 Sucrose is inverted by enzyme invertase that is generally produced by yeast cells
 Sucrose glucose + fructose- C02+alcohol by yeasts and to C02+H2 by
coliforms.
 The organisms are heat sensitive. So the post forewarning contamination is the
major reason.
 This is more common in warmer months. The defect develops very slowly because
of slow growth rate of responsible organisms in the high sugar concentration.
Prevention: -
 Avoid contamination with yeasts
 Avoid contamination of sugar by protecting from dampness and from insects and
Clean sugar conveyers after each days use
 Use concentration sugar syrup in boiling hot water
 Sanitization of all equipment's
 Avoid prolonged exposure to air
 Containers are filled fully giving reasonable allowances for heat expansion
BACTERIAL THICKNENING:
It is the most common defect and is due to Physico-chemical and bacterial origin.
Signs: -
 High bacterial count
 Disagreeable cheesy odor and taste
 Increase in acidity
 When thickened product is diluted with water and heated the curd separates
distinguish from heat or age thickening
 Organisms: - Micrococcus pyogenes, S. aureus, S. albus, B. subtilis and Certain
yeasts
 Mechanism: -
 This is due to the production of rennin like enzyme by organisms, which acts
on milk proteins.
 When sugar contents are high, the growing conditions are rather unfavorable
and thickening proceeds slowly.
Predisposing factors: -
 Contamination with defect causing organisms
 Storage at high temperature
 Low sugar ratio
Public health significance:
 SCM is not a sterilized product either before or after it is canned the

reliance is mainly on
 low moisture content, preservation action of sugar.
 The pathogens are mainly aerobic and anaerobic spore formers. St.
aureus, E.coli and pathogenic yeasts and molds of higher sugar
concentration tolerant ones are also important.
 Preformed staphylococcal enterotoxins are also of greater concern.
Microbiology of DRIED MILKS
 Objectives: -
1. Handling excess milk supply in a dairy factory during the flush
season
2. Optimal conservation of natural properties that are characteristics
of fresh milk
3. Dependable keeping quality of finished product
4. Reduction in bulk to facilitate economic transportation to all parts of
the word
5. Product that can be used in other products like ice cream, infant
foods.
 Dried milk products: - Whole milk powder, Skim milk powder, Infant
foods, Malted milk foods, Dried ice cream mix, Dried whey and
these have the water activity of 0.1 –0.3
 Microbial standards: -BIS and PFA
 Methods:
 The methods are
• Roller drying, Spray drying, Freeze drying, Vacuum drying
1. Roller drying: Involves the application of milk in the form of thin
film over the continuously rotating steam heated metal drum, roller
and dried film is continuously scrapped by stationary knife called
scrapper or doctor blades
2. Spray drying: Consists of atomizing the milk, preferably
preheated and concentrated to form a very minute droplets which
are directed into a large chamber where they mix with a current of
hot air
3.Vacuum drying: Revolving metal drums are enclosed in a vacuum
chamber in order to use reduced temperature for drying
Type of flora:
 This include microbes surviving the heating and drying process
and contaminants from air, utensils etc.
 Expected sources:
 Thermoduric flora originating from milk, Species/strains of
bacterial that survive and multiply during drastic processing
conditions
 Residual flora as contaminants from various equipment's
 Post processing contaminants from utensils, air soil
 Contamination of feed tank, Microbes associated with personnel
handling product
Type of flora:
 The common Cocci from enteric source predominately found in

dried products are Enterobacter faceium and Enterobacter faecalis.
 The other organisms: -Coliforms, Salmonellae, Staphylococci,
Haemolytic streptococci, Hafnia, Actinomycetes, and
 Molds such Asperigillus, Mucor, Penicillium.
 Types of Flora
 Dried milks owing to their low moisture content have a long
keeping quality.
 Although the microbes in dried milks can not grow and thus do
not play any direct role in their spoilage their occurrence in
these products is of great significance and serves as an index of
hygienic standards maintained during production, processing
and handling.
 The microbes present may cause defects in derived products
E.g. Processed cheese, reconstituted milk.
 Certain number of microbes even in low number may
constitute a potential hazard. Hence the quality control
personnel are interested in determining both the number and
type of microbes which vary considerably with the method of
manufacture of these products.
 Factors affecting microbiological quality of dried milks and
their control measures :
A. Raw milk: -
Quality of raw milk: -
 Quality of raw milk used directly influences the quality of
finished product. It should have desired properties
 It should be fresh and free from developed acidity
 Activity of enzymes like lipase, protease should be minimum
 Milk should be of good hygienic quality
 It should not contain heavy metals such as copper and iron
 Should be of very good microbiological quality particularly if
low heat powder is to be manufactured
A. Raw Milk (quality)
 The most important organisms, especially thermoduric aerobic
spore formers should be low in number
 Should have low Thermophilic count. There is simple
opportunity to increase in numbers at stages of manufacture
due to their ability to grow under processing conditions
 MBRT should not be less than 3.5hr
 Acidity not more than 0.03%LA than normal
 No objectionable flavor or odor
 To assess general hygiene of milk quality, SPC, Thermophilic
count and coliform count are important .
Clarification / Filtration:
 The plate count may be higher in clarified milk due to
disintegration of bacterial clumps but the effect on microbial
quality is not very significant.
 After each use clarifier should be thoroughly rinsed with ward
water and slime must be brushed off with a detergent.
 The discs should be removed, cleaned and dried.
 Stone formation should be prevented
Separation:
 Skim milk contains fewer microbes since many bacteria are
trapped in the slime.
 However it does not affect the keeping quality. Separator should
be kept clean and sanitized.
Standardization:
 May contribute additional micro flora because of additives but it
does not create any difficulties
Homogenization:
This is an essential step in roller drying because
 to accomplish uniform distribution of fat in WMP
 Facilitate reconstitution of dried milks to fluid milk
-It has appreciable effects on total bacterial count due to breaking
of bacterial clumps. Contamination from homogenizer by
contaminated cooling water past the pistons due to worn or
defective chevron ring.
B. Processing:
1. Pre heating: - Spray drying 61.1-93.3°C, Roller drying 65.6-85°C
but common temperature is 85°C / 10-30 min. It destroy most of
bacteria except heat resistant forms, inactivate milk enzymes
and reduces number of thermoduric.
 Good sanitation, Good raw milk are important in obtaining low
count milk instead of increasing preheat treatment.
2. Concentration: -
 Spray dried milk: - Vacuum pan / Evaporator, concentrated to the
extent of 2:1 or 3:1. There is hardly any chance of aerial
contamination as air enters at a very high temperature. Pre
heating temperature for NFDM of low heat 72°C / 15 sec, high
heat 85°C / 20 min, WMP 88°C or above
B. Processing:
2. Concentration: -
 Roller dried milk: - Pre concentrated to the extent of 3:1 or 5:1.
-Poorly cleaned evaporators / condensers may cause outbreaks of
Thermoduric or Thermophilic bacteria.
-UHT heads in evaporator inclusion gives high quality of powders. 3 to
4 stage continuous evaporators are used, then cleaning should be
carefully done .
- Crucial points are Return loops, Blank ends, Feed control valves,
Vapor ducts
B. Processing:
3. Drying:-
 Direct feed to drying chamber or through feed tank or balance tank.
 Balance tank: -
 Poses microbial hazards as these provide suitable temperature for
growth of microbes and lead to a build up of contaminants
 Staphylococci when present may grow, produce heat stable
enterotoxins that can result in food poisoning out breaks
Mesophiles can grow at this stage
 A high total solid tends to form sludge encouraging microbial
growth contaminating fresh lot.
 Covers should be retained on these tanks and milk level should be
kept as low as possible.
B. Processing:
3. Drying:-
 Spray drying: -
Dryers: - Jet or nozzle type (US type) and Rotary atomizer type
(Europe type)
Drying causes: -
 Instant death of a majority of microbes
 Remaining microbes surviving may die slowly as a result of
oxidative changes
 Some bacteria still may survive because of
o Protective layer of milk solids around them
o Evaporation is always associated with a cooling effect in substrate,
thus bacteria are further protected from high temperature.
o Dry air is not as effective as a bacterial agent as moist air
B. Processing:
3. Drying:-
 Jet spray drier:
 Concentrated milk posses through a high pressure pump to a series
of jets within drying chamber into a conical thin sheet, broken into
droplets and dried instantaneously by contact with hot, dry
incoming air so no possibility of any microbial hazard
 Rotary atomizer:
 It is a spinning disc with holes. Heavy powder falls to bottom, fine
powder is to be recovered using cyclone or fabric filter type, cyclone
recovery is appears to be safest.
 Filtration of air before heating using retentive filter increases the
quality of powder.
B. Processing:
3. Drying:-
 Roller drying: - Atmospheric – run at 148-149°C; Vacuum – run at
100°C
 High temperature destroys bacteria drums reduce the viable
microbes and hence powders prepared by roller drying contain
fewer microbes than spray dried.
 Bacterial content is due to
 Hood used for vapor removal provide a harbor for microorganism
 High humidity in hoods is responsible for microbial proliferation
 Manual contact with powders during recovery is reason for
contamination
 Use disposable plastic gloves
 Mechanical recovery
B. Processing:
4. Instantization: -
 Forming agglomerates of powder to improve the working and
dispersing properties.
 This process increases microbial hazards.
5. Packaging / Storage: -
 Aim is to protect dried milk from moisture during packaging and
storage.
 Dried bacteria die at a greater rate in air in presence of increased
moisture and temperature and die at a slower rate in atmosphere of
Co2, H2 and N2.
 Usually inert gas packaging is used. Filling operation may be source
of contamination
 Storage changes: - Roller dried- decreases rapidly first and
relatively constant after 2 to 4 months and in spray dried –milk the
decrease is less marked. The Spore formers and Micrococci survive
longest
C. HYGIENE:
 This is very important especially in plants with high staphylococci,
yeast and molds and coliforms.
1. Air: This plays a significant role because of a large number of air and
product contact points during spray drying, Instantization and
packaging.
 Measures to minimize contamination:
o Air filtration with ultrahigh efficiency air filters to remove microbes
o Periodic cleaning of filter pads to remove accumulated dust
o Bacterial spray i.e chloromist
o Sanitary conditions in drying room
o Opening of side windows to rectify clogging of conveyer should be
avoided
C. HYGIENE:
2. Caladria/ Vacuum pan: Milk stones become loci for multiplication of
microbes. Circulate acid and alkali and finally sterilize by steam and
chlorine.
3. Pipe lines: Clean thoroughly those sections which do not form part
of circuit eg. Valves
4. Concentration tanks
5. Packaging rooms
6. Storage of containers
7. Personnel hygiene
Defects in Dried Milk
 Dried milk has longer shelf life as compared to other products.

No specific defects can be attributed to microbes until it is
reconstituted because low moisture level.
 On contrary there is decrease in number of bacteria during
storage.
Storage on microbial quality: -
 Microbial population decrease because of low water activity.
 A rapid fall in number of bacteria is observed followed by the growth of
yeast and molds at high humidity.
 Micrococci survived at 5-15% relative humidity.
 There is decrease in fecal streptococci, psychrotrophs caseolytic and
lipolytic organisms at 18-25°C and at RH of 60-65%
 S. aureus, Sal. newport survival was greatest at water activity of 0.22 and
decreased with increased water activity
 Pressure of air has adverse effect on survival particularly of staphylococci
and 1.5% oxygen in nitrogen was nearly as destructive as air.
 For 99% decrease in count it takes 18 weeks for staphylococci, 9 weeks for
salmonellae.
 Coliforms were considerably reduced after 3 months
 Staphylococci were resistant with 50% survival after 3 months
 Thermophilic count was decrease by 11-25% at 10-20°C after 3 months.
 Reconstitution on microbial quality: -
-This is used for propagation of starter cultures and used for
preparation of cultured milk products but Inhibitory substances –
antibiotics, sanitizers, detergents carried from initial raw milk
may in inhibit starters
 If reconstituted milk is held for 24hrs, there will be little
increase in total count at 15.5 0C, two fold increase at 22.2 0C
and clotting of milk at 37 0C by St. faecalis.
 Factors affecting
• Temperature of reconstitution
• Type of water
• Sanitary condition of utensils
• Consumer practices
• Unboiled water may be a source of coliforms and B.cereus
Public health significance
 Important organisms are Staphylococcus aureus, Salmonella
 Because they cause food poisoning outbreaks, may carry these
organisms to final product in which dried milk is a constituent.
 Roller dried milk is safe and reported to be free from pathogens.
Spray dried milk may contain pathogens and the possible
sources of these organisms are
 Survival during processing
 Milk handlers, dirty equipment's, infected powder water
 External agencies – dust, rodents, birds, vermin etc
 Air with infected powder
 Contamination during drying, cooling, instantizing, packaging
and transportation
Public health significance
 Important organisms are Staphylococcus aureus, Salmonella
 Staphylococci:
 A small portion may survive spray drying mainly due to contamination
from plant environment as pasteurization kills those present in milk.
 Preformed enterotoxins are not destroyed even at boiling and
responsible for food poisoning outbreaks
 Salmonella: -
Sources: personal contact, air water.
 Two important factors for survival of salmonella are product
temperature and particle density.
 Death rate is enhanced by high fat content. These are not killed
completely by spray drying.
 Storage at 45°C and 55°C has lethal effect on Salm. typhimurium,
Sal.thompson
 Storage at 25 and 35°C also led to reduction in population in 4 to 8
weeks
Survival of pathogens
 The chances of survival increased with Modern processing
and Modern packaging.
 Modern processing: - Modern processing is to improve flavor and
solubility. So vulnerable to contamination and growth of
contaminants besides enhancing the possibility of survival of
microbes.
o Milder heat treatment during spray drying compared with roller
drying
o Instantization
o Increased production with less hygiene
 Modern packaging techniques are designed to prevent chemical
deterioration allow longer survival of microbes including
pathogens
 Quality control tests:
-SPC, Coliforms, Thermophilic bacteria, Spores ,Staphylococci,
Salmonellae, Hemolytic streptococci,Yeast and molds
 Quality control programme :
This is to detect faults and produce a product of high quality. The testing
schedule should be
 If coliforms are present in concentrated milk ……..test by swab count of
pipe line especially
 The bypass between two calandria
 The bypass and valve near balance tank
 Concentrated milk tank
 If coliforms are present in powder………..test hygiene of
The pipelines from concentrated milk tank
- The inlet pipe of atomizer
-Personnel helping to prevent clogging in the conveyer
Quality control programme (contd..)
 Air in condensing and drying room containing 10 org/min/sq ft……use
chlorine spray at a concentration of 500-1000 ppm
 Presumptive coliform test should indicate absence of coliforms in 5-6
ml of water.
Types of Dried Milk (Infant Milk
powder)
 Types of Infant milk foods,
 Processed cereal infant foods, Special infant foods
(Humanized milks)
Microbial quality:
 Microbial quality is of great concern because of vulnerable,
sensitive group of consumers.
 Product is not of uniform quality because of variation in
Types of Dried Milk (Infant Milk
powder)
Microbial quality:
 Microbial quality is of great concern because of vulnerable,
sensitive group of consumers.
 Product is not of uniform quality because of variation in
quality of milk and conditions manufacture.
 Spray drying - Milder heat treatment than roller dried milk
 Roller drying - Severe heat treatment
 Instantization - Wetting and redrying may survive as additional
opportunity for contamination.
 Organisms used as index of hygienic quality are Staphylococci,
B. cereus, Coliforms and Enterococci
Micro flora of infant foods:
 The following isolates were obtained by different

workers
Public health significances:
 Infants are highly susceptible consumers, so requirement of high
microbial quality
 Initial quality of milk free from pathogens is very important
 Toxic metabolites lead to vomiting, giddiness, nausea,
dehydration and even to death
 Staphylococcal enterotoxins are of greater concern. Minimum
population required is 10^6/ml. Toxin production in low count
milk is faster in poor quality milk, competition with other fast
growing organisms and changes produced by them – acidity,
depletion of nutrients – may inhibit toxin production by
staphylococci. Ideal place is concentration tanks. Enterotoxins
are not destroyed even after boiling for 15 min
 The other important organisms are Salmonellae, Coliforms,
Clostridia, Bacillus
Organisms of Public Health
Significance
Staphylococcus aureus: -
 They are Gram-positive cocci, non-motile, Negative oxidase.
When low number of staphylococci are expected –
 then follow enrichment procedure usually in broth containing
high salt concentration then sub cultured on Baird Parker’s
medium.
 This medium is not very selective in case of dairy products.
The Principle lies in reduction of tellurite and production of
egg yolk reaction producing typically black colonies.
 Identification:
1. Utilization of glucose anaerobically to produce acid but
variations are observed.
2. Produce acid aerobically from glycerol in the presence of
erythromycin.
3. Sensitivity 0.4 micro gram / ml. Sensitivity to lysostaphin
Organisms of Public Health
Significance
Contd..
 Supplemental tests :
1. Coagulase test,…most widely used for milk and milk products but
this enzyme may also be produced by S.intermedicus,
S.chromogenes, and S.hyicus
2. Thermonuclease test: almost 99% of coagulase positive
staphylococci produce this enzyme
 Detection of entertoxins: Molecular weight is between 28,000 to
34,000 and can withstand 100◦C for 30 min. The toxins are
detected by
a) Radio immuno assay (RIA)
b) Enzyme linked immunoSorbent assay (ELISA)
c) Reverse passive haemaglutination assay (RPHA)
Organism of Public Health
Significance
Contd..
 There may not be correlation between number and detectable
enterotoxins because
a) The organisms killed during processing after producing of toxins
b) Small number of viable organisms gained access to the finished
product
c) Outgrowth of organisms after toxin production
 Salmonella:
 These are G –ve asporogenous and facultative short rods, motile
by peritrichus flagella, Grow best between 5.5 to 45 ◦C and can
with stand aw of 0.945 to 0.999.
 For the detection the preliminary enrichment is recommended in
tetrathionate broth followed by plating on selective media like
Bismuth sulfate agar which gives rise to black colonies with
metallic sheen colonies.

Dairy Microbiogy Notes PPT11

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Dairy Microbiogy Notes PPT11

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DAIRY MICROBIOLOGY

 when milk fat is concentrated into a fraction of the original

 NS – Not less than 18% Fat

Half and Half Cream: It is usually used in tea and coffee.

In coffee, pouring on fruit and desserts,

Pouring/spooning cream for desserts,

Aeration for applications including

Contains no less than 35% milk fat (Cream has been

Used in the classic English cream tea

Production of Transport to Storage in

 If spore content is high (>100 ml-1) it may be worthwhile to

• Separating the milk at slightly higher fat content than

 Destruction of all pathogens

 The protective effect of fat on the bacteria

 Tyndallisation i.e., heating at 100°C for 30 min is also

 Cartons whether of paper or plastics, are of good hygienic

 Bulk quantities are transported in a stainless steel tanker

 Intermediate quantities are distributed in steel or aluminum

 Common water borne organisms

 This is least objectionable method.

 Exit from Pasteurizer

 Keeping quality is of 14 days with the cream held at

 From the point of keeping quality in cream bacteria

Plate or colony count Cells or clumps forming colonies under the

Modern refined instrumental Production of metabolites or changes in

 Butter may be defined as fat concentrate obtained by

 Sour cream butter

 Cream micro flora impairing quality of butter

Cooling 20-22°C Cooling 5-10°C

Churning (temperature. 9-11°C)

(2-2.5% of fat) salting and working

 Butter cultures contain lactic acid producers such as L.

-Cool cream to 19°C -Cool cream to 6-8°C

 Neutralization changes the pH of the cream to levels of

 Heat treatment used is high >71.1°C/30min and Usually

 Working of butter causes little quantitative change in micro

Temperature of Very good Keeping Good keeping Poor keeping quality

15 5 weeks 20 days 3 days

10 2 months 4 weeks 1 week

0 3 months 6 weeks 1-4 weeks

-12 9 months 6 months 1-3 months

-25 12 months 9 months 3-6 months

Satisfactory <100 <10

Doubtful 100-1000 10-100

Unsatisfactory >1000 >100

 Flavor imparted to butter depends on the amount of flavor

E.g. Ps.putrefaciens effectively inhibited by the addition of 5 or

 In unsalted butter favorable temperature may permit the

1. Lack of delicate and characteristic flavor. Have somewhat

 The normal flavor of the butter should be fresh, clean flavor,

 Flat or insipid flavor: - occurs in fresh butter and is due to

 Medicinal Flavor: - is due to the result of

 Putrefactive Taint: - This is the most common bacterial

 Low bacterial counts in the raw materials

 Malty Flavor: - is due to malty variants of

Acid Coagulation Paneer (non fat)

Whole Milk Condensing Kulfi

Peda Burfi Lal Mohan

 Processing conditions are not hygienic at halwais, and