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Isolation of Lactobacillus acidophilus

Introduction:
The microorganism that I have selected is Lactobacillus acidophilus. "Lacto" means milk and
"bacillus" means rod like. Acidophilus means it is acid loving. It is a member of the Kingdom
Eubacteria.

 Scientific name: Lactobacillus acidophilus


 Kingdom: Bacteria
 Phylum: Firmicutes
 Class: Bacilli
 Order: Lactobacillales
 Family: Lactobacillaceae

General characteristics:
 It is a gram positive bacteria
 L. acidophilus produces the acids that make food taste sour
 It is rod shaped coccobacilli
 It is homofermentative specie.
 It ferments sugar into lactic acid
 It is non-spore-forming bacteria.
 It is facultative anaerobic or microaerophilic,
 It is very acid-resistant and tolerates bile quite effectively
 It is well known that Lactobacillus acidophilus is catalase negative

Cultural characteristics:
Part of the normal flora of mouth, intestinal tract and vagina of humans and other warm-blooded
animals. Also found in dairy meat, fish, pickled and fermented products
Growth Requirements:
 It grows readily between pH 6.5 to 8.
 It has an optimum growth temperature of around 37 °C (99 °F)
 They can grow in the absence of oxygen
RECOMMENDED MEDIA
For culture: Rich, complex media.

For selective isolation: Universal Beer Agar, MRS Agar and Broth.

For maintenance: Skim Milk Media may be used for storage at -70 degrees C.
Lyophilization may be used for preservation.

INCUBATION
Temperature: 30-35 degrees C.

Time: 24-72hr

Atmosphere: Anaerobic or in 10% CO 2 .

Strains
Laboratory: NCFM, 4962, CNRZ216, CNRZ218
Human: HA1, HA2, HA3, HM2, HM6
Pig: PA3, PA12, PA19, P18, P47
Chicken: C1, C2, C3, C7, C11
Isolating Methods:
 Sterilize all the things except sample
 Prepare the media (MRS agar)
 Pour the media into petri plates
 Curd is taken in sterilized flask. Under aseptic conditions curd was serially diluted from
10-1 to 10-14 from this 14 dilutions 10-5 10-7 10-9 are selected.
 For this 3 tubes spread plate technique farther with streak plate technique is done on
MRS medium.
 They are incubated in incubator at 37°C for 24 hours.
 After the period of incubation 3 isolated colonies were grown
Biochemical tests result:
Many test reactions are weak and dependent on the composition of the media and exact cultural
conditions

TEST OBSERVATIONS
Arabinose No fermentation

Sorbitol No fermentation

Maltose Fermentation (acid production)

Lactose Fermentation (acid production)

Sucrose Fermentation (acid production)

Glucose(acid) Fermentation (acid production)

Glucose (gas) No gas formation (no bubble in Durham tube )

Colony morphology:
MICROSCOPIC APPEARANCE
Gram Stain: Positive.

Morphology: Long, slender rods to short, coryneform coccobacilli.

Size: 0.5-1.2 micrometers by 1.0-10.0 micrometers.

Motility: Non-motile, rarely showing motility by peritrichous flagella.

Capsules: None.

Spores: None.

Colony appearance:
Typically, colonies on agar media are usually 2-5 micrometers, convex, entire, opaque, and
without pigment.
Lactobacillus acidophilus (ATCC ® 4356) colonies growing on Lactobacilli MRS Agar (Cat. no. G117). Incubated in CO 2 for 72 hours at
35ºC.

Importance of Lactobacillus acidophilus


 L. acidophilus occurs naturally in the human and animal gastrointestinal tract and mouth.
 Some strains of L. acidophilus may be considered to have probiotic characteristics.
 Some strains are commercially used in many dairy products, sometimes together
with Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus in the
production of acidophilus-type yogurt, or acidophiline
 Lactobacillus acidophilus has also been used to treat lactose intolerance, Crohn's disease,
overgrowth of bacteria in the intestines, or vaginal yeast infections caused by antibiotics.
 It competes against or "antagonizes" an array of pathogens including Escherichia coli (E.
coli, the cause of "Montezuma's Revenge"), Staphylococcus aureus and Salmonella
(common causes of food poisoning), Candida albicans (yeast infections and syndromes),
and other pathogens such as Shigella, Clostridium, Listeria, and Helicobacter species
 Life without Lactobacilli is unimaginable: They are used in the production of yoghurt,
cheese, chocolate, sourdough bread, sauerkraut, pickles, kimchi (a traditional Korean
pickled dish), beer, wine, cider and many other fermented foods.
 Lactobacilli are also important in animal feed (silage) production.
 They produce lactic acid, lower the pH and thereby inhibit growth of bacteria

References:
https://microbewiki.kenyon.edu/index.php/Lactobacillus_acidophilus

https://www.ijppsjournal.com/Vol6Issue1/7982.pdf

https://catalog.hardydiagnostics.com/cp_prod/Content/hugo/LactobacilliMRSAgar.html

https://catalog.hardydiagnostics.com/cp_prod/Content/hugo/Lactobacillus.htm

http://bacmap.wishartlab.com/organisms/1292

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