Microbiological Technique

Assignment
(Practical)
Topic: 1. Identification of Lactobacillus Acidophilus
. Total !iable "ount
#ubmitted to: Madam $areeha
#ubmitted b%: Pharmacolog% &roup
(M.phil ' Ph.()
)
rd
#emester
The *ni+ersit% of !eterinar% ' Animal #ciences
Total !iable count

Total Viable Count (TVC) gives a quantitative idea about the presence of microorganisms
such as bacteria, yeast and mold in a sample. To be specific, the count actually represents the
number of colony forming units (cfu) per g ( or per ml) of the sample.
This involves counting the colonies produced by viable cells under favourable growth
conditions. In pourplate method, an aliquot of suitably diluted sample is mi!ed with nutrient
agar at a temperature where it is liquid. Then the mi!ture is poured into petridishes and allowed
to set.
"lternatively an aliquot of the sample is spread over the agar surface of a #etridis using a
sterile spreader. $embrane filters can also be used to determine the bacterial numbers. In this
method cells are filtered onto membrane filter which is then placed over nutrient agar surface.
,esults
Identification of Lactobacillus Acidophilus

#tud% -ac.ground:
Lactobacillus acidophilus and other lactic acid bacteria are important in the fermentation of
many foods from dairy products to fruits and vegetables. %ermentation occurs when bacteria
brea& down sugars and carbohydrates to produce alcohol, carbon dio!ide and lactic acid.
These byproducts are responsible for the unique taste of fermented foods and help preserve
and increase palatability.
L. acidophilus can also be used as a probiotic or living organism, which upon ingestion in
certain numbers, e!ert health benefits beyond inherent basic nutrition. 'actic acid bacteria use
lactose as their main source of carbon to produce energy. 'actic acid bacteria use energy to
transfer lactose (main sugar of mil&) through their cell membrane. The lactose is metaboli(ed to
lactic acid and in some species also acetic acid, ethanol and carbon dio!ide. 'actic acid
bacteria that only produce lactic acid as an end product are called homofermentative) those
that also produce acetic acid, ethanol and carbon dio!ide are termed heterofermentive. The by
product of this reaction is energy that the bacteria use for growth. The end products of
fermentation ultimately change taste and te!ture of food.
L. acidophilus is a member of one of the eight main genera of lactic acid bacteria. The genera,
Lactobacillus, Streptococcus, Lactococcus, Leuconostot, Bifidobacterium, Carnobacterium,
Enterococcus and Sporolactobacillus can then be divided into species, subspecies, variants
and strains.
Lactobacillus are able to live in highly acidic environments of p* +, or lower. This p* is well
below the p* other lactic acid bacteria can live in and because of this, 'actobacillus is
responsible for the final stages of fermentation in products.L. acidophilus is probably the best
well&nown species of Lactobacillus . L. acidophilus is naturally found in the human and animal
-I tract, mouth and vagina. It can also be found in certain dairy products and free(e dried in
capsule and tablet form in grocery and health food stores.
Morphology: The overall shape of '. acidophilus is one that
is rod shaped. This bacterium occurs in small chains and is
usually .., to ../ micrometre (0 mm10.
2
metre) across by 3
to 4 mm in length. It is also important to note that '.
acidophilus is nonsporeforming.
This is the assignment of our class wor& as a group for the identification of 'actobacillus
acidophilus in laboratory.
#teps:
1. Lactobacillus acidophilus was taken present in powder form in probiotic Sachet named
Acotec containing Bifidobacterium, Lactobacillus acidophilus, streptococcus
thermophile.
2. Powder was dissolve in sterilied distilled water.
!. Solution was poured on agar media plate under aseptic conditions.
". Plates were incubated at in incubator for 2" hours.
#. $e%t da& plates were removed from incubator and mi%ed growth appeared.
'. (ulture was purified b& streak method on another media plate under aseptic conditions
and allowed to grow for 2" hours in incubator.
). Pure growth was appeared on media plate, Lactobacillus acidophilus was identified
according to its morpholog& and further scheme was followed to identif&.
#cheme to identif% Lactobacillus Acidophilus
Bifidobacterium,
Lactobacillus acidophilus,
streptococcus thermophile
Acid
Fast
Staining
Carbolfuchsi
n
5ed color
'actobasillus 6pp.
Catalase Test
7ubble
formations,
negative
'actobacillus
"cidophile confirmed