The propagation
and preparation of LAB starter cultures is one of the most important operations
in a dairy plant required for manufacture of fermented products. Since the
quality of a starter has a direct bearing on the quality of the final product,
starter must be pure, active and in good shape. The good quality starter
cultures could only be prepared by employing certain essential factors such as
selection of good quality milk, necessary heat treatment of milk, well cleaned
and sanitized utensils, optimum amount of inoculum, aseptic transfer of
culture, incubation at appropriate temperature and time, cooling and properly
storing till its use.
i.
Selection of milk
The milk selected
for preparation of starter must be from healthy animals that are secreting
normal and clean milk, free from inhibitory substances. The abnormal milk,mastitic
milk and milk with unusually high lipolytic activity and low in total solids should
not be used for preparation of starter cultures as well fermented products.
ii.
Heat treatment of milk
Milk to be used
for starter propagation should be given adequate heat treatment to inactivate
the normal germicidal effects and also to destroy the majority of the microorganisms
present as normal flora or contaminants. To achieve best results milk should be
heated to minimum of 71.1oC for 30 min. Sometimes
sterilized milk is recommended for starter propagation. However, more drastic
heat treatment results in the inhibition of culture and the texture and body of
the curd formed is soft and the scorched flavour developed therein hinders the
accelerate assessment flavoring and aroma development. The steaming and boiling
of milk for 30-60 min for mother
culture is
preferred. After heat treatment, immediately milk is cooled to incubation temperature
to minimize the physico-chemical changes in it.
iii.
Containers/ Utensils
Glasswares,
stainless steel, aluminium, enamelled wares are the most appropriate material
for starter propagation. The copper and copper alloys should not be used.The
surface of containers should be smooth and free from crevices. The container should
be thoroughly cleaned and sterilized by steaming for at least 30 min at 100oC.
iv.
Level of Inoculum
The level of
inoculum used for propagation of starter cultures may also affect the performance
of the starter. This depends upon the characteristics of starter culture,activity
and condition of the culture at the time of inoculation, time and temperature of
incubation employed. Normally 0.5 to 2.0% inoculum is used for starter preparation.
The amount of inoculum may vary according to type of starter culture.
v.
Culture Transfer
The starter
culture should be properly and aseptically transferred avoiding all the changes
of contamination. The strict aseptic conditions are needed to avoid aerial
contamination with bacteriophages, molds and other undesirable microorganisms.For
this purpose, a separate culturing room should be constructed and adequately equipped
with laminar flow, UV lamp and provision for spraying cleaners and sanitizers.
For transfer of cultures, all the accessories required should be thoroughly cleaned
and sterilized.
vi.
Incubation period
Normally, after
inoculation the majority of milk starter cultures are incubated at 21.1oC
to 30oC and time of incubation is determined depending upon the inoculums
size and the activity of individual culture. In general incubation time of
14-16 hours appears to be ideal when the inoculum level used is 1 percent. The
maintenance of proper temperature is very important and hence, it should be
carefully monitored.
vii.
Cooling
After attaining
the desirable growth of the starter culture, it should be immediately cooled to
stop further growth so that it should be in sound and active condition for further
use. The cultures are stored under refrigeration including mother cultures until
further use.
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