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Lactic Starter Cultures And Related Products

Culture Product
Cottage cheese, buttermilk, sour cream,
Lactococcus lactis subso Lactis
cheddar, soft and semisoft cheeses, Gouda,
Lactococcus Lactis subsp. cremoris
blue vein cheese, other cheese.
Lactococcus Lactis subsp. lactis biovar Cottage cheese, buttermilk, sour ceram,
diacetylactis senisoft cheese, cheddar
Yogurt, Mozazrella, Emmamentaler, Gruyere,
Streptococcus thermophilus
Swiss, Hard italian cheese

Lactobacillus delbrueckii subsp. bulgaricus

Yogurt, Mozzarella, Emmamentaler, Gruyere,
Lactobacillus delbrueckii subsp. lactis
Swiss, hard italian cheese, kefir, koumis
Lactobacillus lactis

Lactobacillus acidophilus Acidophilus milk, yogurt emmentaler,

Propionibacterium shermanii Gruyere, Swiss, Gouda
Bifidobacterium ssp Probiotic milk and cultured milk products
Sumber : Kosikowskli et al

Bakteri pembusuk pada keju dihambat oleh efek asam dan garam, rendahnya kelembapan dari
keju. Spores of clostridium tyrobutyricum in the milk used for the manufacture of Emmentaler,
Edam, and Gouda can survive the heat treatment used for cheese milk and cause late gas formation
(blowing defect) and related of flavors during ripening. Thermoduric streptococcus thermophilus
can use flavor defect in Gouda cheese (45). The presence of heat stable enzymes (from
psychrotrophic bacteria) can be detrimental to the quality of both fresh and ripened cheese
(11,12,45) by causing bitter or rancid flavors and by impairing the coagulation properties of the
milk. Fresh cheeses, such as cottage cheese and other high mousture cheeses, may be subject to
spoilage by gram negative psychrotrophic bacteria (Pseudomonas, Falvobacterium, or
Alcaligenes), coliforms, and yeast and molds that eneter as post pasteurization contaminants (10).
Most hard ripened cheese are not subject to gram negative spoilage though coliform contamination
has been associated with the gassy defect in cheese making (for example , cheddar) (41). Ripeden
cheese is prone to surface growth of yeast and molds, particularly if exposed to atmospheric to the
starter cultures, culture failure, and undesirable secondary flora (45).
Cheeese Secondary Flora
Soft (surface ripened)
Camembert Yest
Brie Penicillium caselolum
Caephilly Lactobacilli
Limburger (surface ripened) Yeast, Brevibacterium linens
Blue vein
Roquefort Penicillium roqueforrrti, Yeast, Micrococci
Cheddar Lactobacilli, pediococci
Emmantaler Propionibacterium shermanii, Group D
Gruyere Proprionibacterium shermanii, Group D
streptocci, Yeast, Coryneforms,B.linens

Microbial competititon, reduced water activity, arganic acids, and a low pH generally limit
the growth of pathogens in cheese. A slow starter culture (due to bacteriophage, antibiotics, etc).
Can allow growth of bacteria related to foodborne illnesses such as staphylococcus, Salmonella,
Listeriaa, and enteropathogenic E.coli (6, 37, 52). Which enter with raw milk or as post
pasteurization contaminants. Number of S.aureus will normally decline during the ripening stage,
but if sufficient numbers (>107 per mL) are reached during cheese making, enterotoxin may persist
in the cheese. Salmonella spp, survive beyond the ripening period (6) with the potential to cause
infection at relatively low doses (13). Most but not all, enteropathogenic strains of E.coli are
inactivated ap pH <5,0 (32) although in low acid, semisoft, surface ripened cheese, fecal coliforms
are commonly found (6). Post heating contamination or the use of contaminated raw milk can be
a source of L.monocytogenes, as was implicated in an outbreak involving a low acid Mexican style
cheese (16). Listeria is capable of surviving in cheddar, camembert and cottage cheese (69)
although growth is limited because of the low pH of most cheeses. Biogenic amine formation in
fermented dairy products has been reported by many workers. Edward and Sandine (19), reviewed
the occurrence of biogenic amines in cheeses and discussed the public health significance of the
amines. Histamines produced by the decarboxylation of histidine, and other biogenic amines such
as tyramine and tryptamine cause abdominal cramps, diarrhea, nausea, headache, palpation,
tingling, and flushing, etc. medical intervention is unnecessary, with most symptoms disappearing
a few hours after onset. Biogenic amines also affect some antidepressant drugs by counteracting
monoamine oxidase inhibitors. Enterocci and certain other lactic acid bacteria were implicated in
the production of biogenic amines in fermented dairy products.

Yogurt and Fermented Milks

Yogurt, fermented milks (buttermilk) and cultured cream (sour cream) are unripened,
cultured dairy products. They are generall ready for consumption with the minimum processing
after development of the desired acidity through a lactic acid fermentation. Yogurt fermentation
involves a mixed culture of S.thermophilus and Lactobacillus delbrueckii subsp. bulgaricus, which
are thermophilic in nature, while Lactococcus spp and Leuconoctoc spp are normally used in
cultured milks and sour cream. The more exotic cultured milks (e.g., kefir, koumis) are derived
from mixed fermentations involving yeast, Lactobacillus acidophilus may be used in the
production of bith yogurts and fermented milks (41).
In a normal fermentation, a fina pH of <4,5 is developed in cultured milk products. This
low pH generally prevents the growth of most spoilage and pathogenic organisms, although
interference with acid development may allow growth of the most spoilage and pathogenic
organism, although interference with acid development may allow growth of undersirable
microorganisms. Microorganisms that cause deterioration of fermented milk product can enter
products through poor sanitation techniques or can be introduced by the addition of flavoring
materials. Yeast and molds, which tolerate the lower pH, arethe more predominant organism
involved in the spoilage of cultured milks. Bacillus subtilis and B.cererus can cause bitter flavors
if large numberss survive pateurization. Coliforms, if present, decline rapidly after manufacture of
yogurt, although they may survive in cultured buttermilk and sour cream (52).
Fermented dairy products normally contain high numbers of starter microorganisms or
secondary ripening flora, making total counts insignificant except in products that are heated to
inactivate added cultures. Yeast and mold count and coliform counts may be used as indicators of
adequacy of processing santation with some fermented products. Howefer , it needd to be
determined if the source of the contamination is poor sanitation or cantaminated flavoring
materials that might have been added to the products. Enterococci are probably better indicators
of improper sanitation than coliforms because coliforms are quite sensitive to high acid condition
prevelant in fermented products and might not be recovered on selective media because of acid
injury. Counts of psychrotrophic bacteria are useful for cottage cheese and similar products that
are subject to spilaoge by pychrotrophic bacteria.

Recommended methods
1. Yeast and Molds Counts
2. Coliforms Counts
3. Psychrotrophic count
4. (Cottage cheese)
Other methode for specific determinations
1. Staphylococcus Enterotoxin/ Thermonuclease
2. For individual pathogens such as salmonella, campylobacter, Listeria, and yersinia consult
appropriate chapters of this book.

Acid Producing Bacteria In Dairy Foods

One of the most important groups of acid producing bacteria in the food industry is the
lactic acid bacteria. Members of this group are gram positive, nonsporulating cocci or rods,
dividing in one plane only, with the exception of the pediococci. They are catalase negative (with
the exception of the some pediococci, which either form a pseudo catalase or incorporate
preformed hemin supplied exogenously, into a catalase molecule). The organisms are usually
nonmotile and are oblgate fermenters, producing mainly lactic acid and sometimes also volatile
acids and CO2. They are subdivided into the genera lactococcus, Leuconoctoc, Pediococcus and
Lactobacillus. The homofermentative species produce lactic acid from available sugars, while the
heterofermentative types produce, in addition to lactic acid, mainly acetic acid, ethanol, CO 2 and
other components in trace amounts. Lactic acid bacteria are widespread in nature and are best
known for their activities in major foods such as dairy, meet and vegetable products.
Propionibacterium also produce acid and several species are important in the development of the
characteristic flavor and eye production in Swiss type cheese.

Lactic Acid Producing Bacteria

The large number of media proposed for lactic acid bacteria, particularly for streptococci and
lactobacilli, is indicative of the difficulties encountered in growing some strains o these organisms.
The choice of medium is governed to some extent by the particular strains under study and
therefore by product or habitat. The media listed below have merit in the support of colony
development of lactic acid bacteria, but are not highly selective and some such as Eugon agar are
not selective at all. Hence, organisms other than lactic acid bacteria may develop on these media
and produce acid.
While the lactic acid bacteria in general are tolerant of low pH, they can be very sensitive
to other adverse conditions. Samples to be examined for numbers of viable lactic acid bacteria
should not be frozen prior to analyses. Many of the lactic acid bacteria are easily killed or injured
by freezing. If the product to be examined is normally frozen, it should not be thawed and refrozen
prior to microbial analyses since this would tend to increase damage caused by freezing.
Dilution of product with phosphate-buffered diluent for plating can damage lactic acid
bacteria in samples to the point that reduced counts are obtained. Thus, it is best touse sterile 0,1
% peptone water as the diluent since it protects bacteria during the dilution process (31,35,39).
Depending on the product, it may be advantageous to blend the initial dilution for the plate
count to disrupt chains of lactic acid bacteria. This is especially true for many freshly prepared
cultured food products. Blending the initial dilution can produce a more accurate count of the
number of bacteria actually present. Chilled diluent and a chilled blender cup should be used.
These bacteria do not grow well aerobically, although most of them are considered to be
facultative. Thus, it is usually important to pour overlays of the appropriate agar medium onto the
surface of the solidified agar in plates containing the lactic acid bacteria. Al alternative is to
incubate the plates in an environment containing little or no oxygen.
The fastidious nature of lactic acid bacteria restricts them in the environment to where to
wherever carbohydrate, protein breakdown products, vitamins, and minerals occur in ampple
quantity and proportion. Therefore, the greatest natural reservoir for these bacteria is growing
green plants. Enrichment culture of blended plant material added to skim milk supplemented with
0,05% glucose and 0,1% yeast extract, with subsequent plating on appropriate media is a common
isolation procedure.Mundt et al (58), Mundt and Hammer (57) and Sandine et al (70) are often
consulted for the isolation of the lactococci and lactobacilli from plants, including vegetables.
Selective Media For Lactic Acid Producing Bacteria
1. Lactic Agar
This medium was developed to support colony development of lactococci (lactic
streptococci) and lactobacilli. Prepare pour plates of sample as describe using lactic agar. After
incubation of plates, prepare gram stains of individual colonies, examine thesemicroscopocally,
and test for catalase reaction. Gram-positive, catalase negative cocci or rods may be tentatively
considered to be lactic acid bacteria. If further identification is needed, consult sharpe (72).
2. MRS Media
MRS broth was developed by deMan, Rogosa, and sharpe (14) to support the growth of
various lactobacii, particularly of dairy origin. Pediococci and leuconostocs grow
luxuriously in his medium. MRS broth with added agar may be used to prepare pour plates
of sample using MRS agar instead of lactic agar.
3. RMW Agar
this is a selective medium for the cultivation of oral and fecal lactobacilli. Pediococci can
also be isolated using this medium. Since some lactobacilli will not grow on this medium if
incubated aerobically, plates should be incubated in a CO2 enriched atmosphere. The

plates can be placed in plastic bags, flushed 1 min with CO2 , sealed and incubated
at the desired temperature.
4. M16 Agar
This medium was developed to support growth of lactococci (lactic
streptococci) used in cheddar cheese manufacturing in New Zealand.
5. M17 Agar
This medium was developed by terzaghi and Sandine to support the growth of
lactococci (lactic streptococci). It is buffered with β-disodium
glycerophosphate and also is usefu for plaque assay of lactic bacteriophages. P
6. Meugon Agar
7. APT Agar
8. LBS Oxgall Agar