For the best
performance and maintenance milk should be sterilized by heating at 90oC
for 1 hour. The breakdown products of milk by employing heat treatment act as
bacterial growth factors. This is probably one of the reasons why lactic acid bacteria
grow more rapidly in heated than unheated milk. After giving sufficient heat treatment
milk is cooled to 22-25oC and inoculated with appropriate inoculum size.After
inoculation, the culture is incubated at 22-25oC
until clotting takes place, and thereafter it is stored in a refrigerator.
Further, a small aliquot of the culture is restarter inoculated
into a similar container containing sterilized milk for storage of mother culture.
The remaining culture is inoculated into the starter can or vessel containing sterilized
milk. The purity of mother culture however is very essential. The inoculums generally
is added at the rate of 1 percent from a culture having approximately 0.8 percent
acidity. The acidity should not exceed 0.9 percent lactic acid. In case of poor
performance including slow activity, any visible abnormality in behaviour,
flavour,and colour and appearance of the starter, it should be immediately
discarded and fresh starters should be used.
The purity and
activity of the starter culture should be maintained by any means to achieve
desirable fermentation efficiently in the manufacture of cultured dairy
products.The starter culture must contain maximum number of viable organisms
and must be very active under production conditions of the plant. For the
preparation of fermented dairy products like cheese, dahi, yoghurt, etc.
starter cultures are often maintained in milk, heated to 90oC
for 1 hour and bulk starters may be grown in milk held at 90oC
for 30 min. This heat treatment is adequate to kill phages and all other
vegetative bacterial cells.
i.
Preparation of Mother Culture
The preparation of
mother culture is a very important step in the production of bulk starters for
manufacture of fermented milk products at large scale. The mother cultures are
generally maintained in narrow neck polyethylene bottles, which are thoroughly cleaned
and sterilized using steam jets. Sometimes these bottles are sanitized with 0.1
per cent -hypochlorite solution. The milk is pasteurized in these bottles at
72-73oC for 45 minutes and the bottles then submerged in the cooling
water, and thereafter, these milk bottles are stored in a refrigerator for
future use. The milk containing bottle is inoculated by injecting few drops of
the desired culture by means of a hypodermic needle, which can pierce the seals
of caps. Thereafter, the content is incubated at desired temperature in a BOD
incubator.
ii.
Preparation of Bulk Starters
The vessels of
different sizes are used for preparation of bulk starters. These vessels are
closed completely air tight after the initial stage of filling with milk. These
cans are filled with milk and heat treated by steam at 72-73oC
for 45 min and cooled to incubation temperatures. The bulk culture is
inoculated and incubated in the same manner as the mother cultures.
iii.
Continuous Starter Production
Recently, this
culturing technique has stimulated a lot of interest and dairy industry is striving
for the introduction of this technique in the cheese plants with the advent of this
technology, the manipulation and transfer of large bulk of starters are now
avoided.
Many cheese plants
today may use a very huge amount of bulk starters every day.This has been possible
only through continuous cultivation of the starters. The handling of large size
cans for bulk starters is extensively cumbersome and inconvenient.Inspite of
convenience and ease, the continuous starter production suffers from a serious
problem. During the operation of starters, if the starter gets contaminated(attacked
by a phage), the whole bulk and all the equipment get contaminated.Hence, the
control of purity and activity is even more important in continuous starter production
than in a batch method of production.
iv.
Preparation of Master Culture
In case of
preparation of master culture, the method of inoculation is the same as for mother
culture. For this, litmus milk may be used in the polyethylene tubes. Specific care
must be taken to sterilize the chalk before adding to the milk as spores in
chalk may be extremely resistant to heat. Litmus milk previously sterilized in
glass bottles is filled into the tubes using a hypodermic needle. These are now
ready for inoculation with the starter cultures. Such cultures may be
maintained indefinitely with careful checking and testing after three months
interval. Polyethylene containers are preferred over glass tubes and bottles as
these are unbreakable, very light easily portable.
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