Sri Ram Dayal Khemka Vivekananda

Vidhyalaya
Junior College

BIOLOGY INVESTIGATORY
PROJECT

Study of PROBIOTICS and their

Preparation

NAME: SAKTHIVELAYUDHAM. G
CLASS: XII A
ROLL NO:
CONTENTS: PAGE NO.
1) Introduction
2) Isolation of probiotics
3) Commonly used probiotics
4) Applications of probiotics
5) Health benefits on probiotics
6) Conclusions
7) Bibliography

INTRODUCTION
Probiotic are microorganisms, which when consumed, are
believed to provide health benefits to humans and other
organisms. We usually think of bacteria as something that
causes diseases. But your body is full of bacteria, both
good and bad. Probiotics are often called "good" or
"helpful" bacteria because they help keep your gut
healthy. When you lose “good” bacteria in your body (for
example, when we take antibiotics), probiotics can help
replace them. Probiotics are naturally found in your body.
You can also find them in some foods and supplements.
Commonly claimed benefits of probiotics include the
decrease of potentially pathogenic gastro-intestinal
microorganisms, the reduction of gastro –intestinal
discomfort, strengthening of immune system,
improvement of skin function, improvement of bowel
regularity, resistance to pollen allergens, decrease in body
pathogens, the protection of DNA, maintaining the
stomach health when receiving antibiotics, etc. Due to its
immense benefits, it is regarded to be very useful. The
history of probiotics can be traced back to the first use of
cheese and fermented products that were well known to
Greeks and Romans who recommended their
consumption. Later, Henry Tessier found that
Bifidobacteria are dominant in the gut flora of breast-fed
babies. He also observed clinical benefits in treating
diarrhea.

When we start to think about probiotics, there are many

aspects we must consider. Its origin, its first use for our
benefit, its importance in our lives, its synthesis for
common man, spread of its knowledge and improvements
and developments in its overall field of study, synthesis
and understanding. Here, we will be seeing into each of
these fields and aspects. Probiotics has been and is a very
intriguing topic as it has influenced many generations of
people around the globe. That is why we must get to
know about it so we can reap full benefits. Then we will
see how probiotics can be isolated for further scientific
test for its study. Already there has been a lot of research
being undertaken on this topic and there are many kinds
of probiotic, which are being used, already in our
everyday lives without us knowing about them! Then we
will go to the roots of probiotic and see its importance in
shaping many lives in the past all around the world. These
findings are sure to leave you astounded. Then we will
see how and where probiotics can/is being used for the
betterment of human, animal and plant life.

ISOLATION OF PROBIOTICS
The isolation of probiotic organisms can be done
using the following methods and materials: -

1. Sample collection-The dairy samples including

yoghurt, cheese, gorout and shour are collected from
different regions. They must be stored in low
refrigerator temperature (-4°C) to protect normal
micro flora and avoid from contamination and
deterioration.

2. Preliminary screening-the preliminary screening

in Phosphate Buffer Solution with PH 3.0 should
performed for three hours.

3. Isolation of bacteria-Bacteria is isolated from

dairy products by using MRS medium. After
dissolving into MRS broth, they were shaken
homogeneously and were incubated at 37°C for 24 h
in aerobic condition. Finally, the single colony of
bacteria was isolated by observing their colonial
morphology.
4. Identification of isolates-performed by
biochemical and morphological tests such as Growth
test at 4 °C and 15 °C in tubes containing MRS broth
and the fermentation of carbohydrates

5. Determination of antimicrobial activity- Some of

acid and bile resistant isolates was assessed for their
antibacterial activity against main three pathogenic
bacteria using well-diffusion method.
6. Final identification was done using classic
microbiology tests including Gram-staining for
detecting morphology, catalase and oxidase tests,
motility, indole producing, growth at 15°C, and
carbohydrates fermentation (arabinose, fructose,
galactose, lactose, mannitol, salicin, sucrose, and
trehalose) test.
7. All Gram-positive and catalase-negative bacilli
were selected for the assessment of antimicrobial
ability. Antimicrobial effect of isolates was
evaluated by disc diffusion test on MHA medium
plated with three pathogens. For this purpose, fresh
culture of isolates was centrifuged (8000 rpm, 15
min) and supernatants were removed. Blank discs
were inoculated with 40 μl supernatant of each
isolate and were placed on separate MHA medium
inoculated with S. aureus (ATCC-6538), B. subtilis
(ATCC-12711), and P. aeruginosa (ATCC-27853)
strains. Growth inhibition zones of pathogens and
isolated lactobacilli inhibitory ability were assessed
after incubation of all agar media at 37°C for 24
hours.
COMMONLY USED PROBIOTICS

1)Lactobacillus acidophilus – is a gram-positive

bacteria which are used for fermenting sugars into
lactic acid and grows readily at low pH values
(below pH5.0) and at an optimum temperature at
around 37 °C. These strains are commercially used
in various dairy products like yoghurts, etc.
2) Lactococcus lactis – is also a gram-positive
bacteria used extensively in the production of
buttermilk and cheese, pickled vegetables, beer or
wine, some breads. But it has also become famous as
the first genetically modified organism to be used
alive for the treatment of human disease. Other uses
that have been reported for this bacterium include
the production of pickled vegetables, beer or wine,
some breads, and other fermented foodstuffs, such as
soymilk kefir, buttermilk, and others.
3) Sacromyces cerevisiae – is a species of yeast
useful for winemaking, brewing and baking. The
process employed here is top fermenting. The
resulting beers have a different flavor than the same
beverage fermented with lager yeast. The carbon
dioxide generated by the fermentation is used as a
leavening agent in bread and other baked goods.
4) Bifidobacterium - is a genus of Gram-positive,
non-motile, often branched anaerobic bacteria. They
are ubiquitous, endosymbiotic inhabitants of the
gastrointestinal tract, vagina and mouth of mammals
including humans. Different species or strains of
Bifidobacteria may exert a range of beneficial health
effects, including the regulation of intestinal
microbial homeostasis, the inhibition of pathogens
and harmful bacteria that colonize or infect the gut
mucosa and the modulation of local and systemic
immune responses.
5) Saccharomyces boulardii: Saccharomyces
boulardii lyo has been used as a possibly effective
aid in preventing the recurrence of diarrhea caused
by Clostridium difficile (C. difficile). However, this
product may not be effective in treating the first
episodes of this type of diarrhea. Saccharomyces
boulardii lyo is also possibly effective in treating
"traveler's diarrhea," diarrhea in people with HIV,
and stomach ulcers caused by Helicobacter pylori.
7) Lactobacillus casei: Helps control diarrhea, has
potential anti-inflammatory effects on the GI and
aids in relieving antibiotic-associated diarrhea.
Produces lactic acid to lower the pH of the gut,
impeding the growth of harmful bacteria. Lives in
the mouth and intestines of both infants and adults.
8) Lactobacillus reuteri: Provides strong protection
against infection and helps maintain a healthy
immune system. Treats and prevents diarrhea. Helps
relieve colic. Releases a substance capable of killing
bacteria, yeast and fungi, making it popular for
vaginal infection support against candida, UTIs.
9) Streptococcus thermophiles: One of the most
useful strains in the commercial food industry. True
starter strain for making yogurt (used in making
cheeses too). Ferments milk sugar (lactose) that
turns into lactic acid, which is effective at preventing
lactose intolerance and also lowers the pH of the
yogurt preventing the growth of harmful bacteria
causing food poisoning. Keeps micro flora of
intestines balanced. May have benefits for
chemotherapy patients.
10) Lactobacillus helveticus: Exerts antimicrobial
activities against pathogens, helps reduce symptoms
of lactose intolerance by breaking down lactose,
helps to prevent and reduce diarrhea, may have
implications on lowering cholesterol and blood
pressure as well as help with calcium absorption.
Recent studies have also shown this strain to be
beneficial in supporting a balanced mood and
healthy management of stress and anxiety. Lives in
the intestines.
 APPLICATIONS OF PROBIOTICS
1. Dairy products – Probiotics are added in most
commonly consumed dairy products such as curd,
yoghurt, cheese, shrikhand, etc. These help add
benefits to our daily consumption so as to initiate a
regular intake of these good bacterium i.e.
probiotics.
2. Confectionary- Even the daily bread that you
consume has probiotics in it. They are also used in
chocolates, biscuits, cookies and cakes. This ensures
goodness with every bite of joy.
4. Health supplements – In case a person still misses
out on his daily dose of probiotics, there are many
health supplements that incorporate probiotics in
them and also probiotic supplements are available
separately too. Fermented probiotic milk based
drinks are also getting a lot popular nowadays.
These not only provide refreshment but also energy
and an adequate dose of probiotics.

5. Therapeutic – They help in improving intestinal

tract health, enhancing the immune system,
synthesizing and enhancing the bioavailability of
nutrients, reducing symptoms of lactose intolerance,
decreasing the prevalence of allergy in susceptible
individuals, and reducing risk of certain cancers.

6. Animal feed – The feed for swine, beef cattle,

dairy cattle, sheep, poultry, turkeys are seen that
sufficient quantity of probiotics are contained in
them.
HEALTH BENEFITS OF PROBIOTICS
1. Immune system modulation- Strengthening of
nonspecific and antigen-specific defense against
infection and tumors .Adjuvant effect in antigen-
specific immune responses. Regulating/influencing
Th1/Th2 cells, production of anti-inflammatory
cytokines. Decreased release of toxic N-metabolites.
2. Decreased detoxification/excretion of toxic
microbial metabolites - Increased Bifidobacterial
cell counts and shift from a preferable protein- to
carbohydrate-metabolizing microbial community,
less toxic and for putrefactive metabolites,
improvements of hepatic encephalopathy after the
administration of Bifidobacteria and lactulose.
2. Crohns disease - Reduction in proinflammatory
cytokines including TNFα, reduction in the number
of CD4 cells as well as TNFα expression among
intraepithelial lymphocytes.
3. Caries gingivitis-Reduction in gingivitis by
L.reuteri, affects on streptococcus mutants,
colonization of the teeth surface by lactobacilli Less
carries after the ingestion of living or oral
vaccination with heat-killed lactobacilli
4. Small bowel bacterial overgrowth-Lactobacilli
influence the activity of overgrowth flora,decreasing
toxic metabolite production Normalization of a small
bowel microbial community Antibacterial
characteristics.
5. Anticolon cancer effect- Anti mutagenic activity
Detoxification of carcinogenic metabolites
Alteration in pro-cancerous enzymatic activity of
colonic microorganisms Stimulation of immune
function Influence on bile salt concentration.
6. Hepatic encephalopathy- Competitive exclusion
or inhibition of urease-producing gut flora
7.Neutralization of dietary carcinogens - Production
of butyric acid neutralizes the activity of dietary
carcinogens
8. Antihypertensive effect- Bacterial peptidase
action on milk protein results in antihypertensive
tripeptides Cell wall components act as ACE
inhibitors
9. Urogenital Infections-Adhesion to urinary and
vaginal tract cells Competitive exclusion Inhibitor
production (H2O2, bio surfactants)
10. Allergy- Prevention of antigen translocation into
blood stream Prevent excessive immunologic
responses to increased amount of antigen stimulation
of the gut.
CONCLUSIONS
We have seen that probiotic maintains gastro
intestinal tract health and functionality. It can be
used for therapeutic purpose such as colitis,
inflammatory bowel disease, and diarrhea in
addition to lengthening the quality of life. It can be
also used as a supplement in the diet of all stages of
human life beginning from child to old age.
Following ban on antibiotic, probiotic is
recommended as feed additive in livestock for
increasing the productivity and product quality.
------------------THANK YOU---------------------
BIBLIOGRAPHY:
NCERT text book
Practical manual
www.icbse.com
https://academic.oup.com/femsle/article/
334/1/1/579191
https://www.ncbi.nlm.nih.gov/pmc/articles/
PMC1482314/
https://www.omicsonline.org/the-benefits-of-
probiotics-on-human-health-1948-5948.S1-003.php?
aid=2177