Probiotics are useful microorganisms to fight against pathogenic bacteria in human gut . Commercially bacteria and fungus are being used as probiotic organisms

1. Seminar topic :-probiotics
Naveen Chaudhary
P.hd scholar
PGIMER ,Chandigarh

2. Probiotics


3. Introduction
History
Properties
Mode of action
Advantages
Application
Probiotic products
Adverse effects
Future aspects

4. Introduction
Origin of life :- Started 3.8 billions years back
The first living things on Earth:- Single-celled microbes lacking a
cell nucleus

5. Introduction
Evolutionary biologists generally agree that humans and
other living originated from the bacterial cells

6. Probiotics
 Probiotics are live microorganisms that are
similar to beneficial microorganisms found in the
human gut. They are also called “friendly
bacteria” or “good bacteria.
 Latin word pro means "for"and the Greek letter
bios means "life".
 The term contrasts with the term antibiotic

7. Definition
 Experts have debated how to
define probiotics
 One widely used definition,
developed by the World
Health(WHO)
 “Live microorganisms,
which, when administered in
adequate amounts, confer a
health benefit on the host ”

8. 8
Antibiotics
Synbiotics
Prebiotics
Probiotics

9. History
 The history of probiotics can be traced to the first use of
cheese and fermented products, that were well known to the
Greeks and Romans who recommended their consumption
 Nobel laureate Élie Metchnikoft suggested Proteolytic
bacteria such as clostridia, which are part of the normal gut
flora, produce toxic substances like phenol and ammonia ,
he called "intestinal autointoxication which would cause
the physical changes associated with old age
Elie Metchnikoff

10.  He postulated that yogurt-consuming Bulgarian
farmers lived longer lives because of their custom.
 Metchnikoff proposed that consumption of fermented milk
would "seed" the intestine with harmless lactic-acid
bacteria and decrease the intestinal pH, and that this would
suppress the growth of proteolytic bacteria.

11. Probiotics facts
 There Are 10 Times More Probiotics Than Cells In
Our Body
 Probiotics Live Throughout Our Entire Body (Not
Just Our Stomach!

12. Appendix Is Not Useless – It Incubates
Probiotics

13. Properties of Effective Probiotics
 Non pathogenic and non toxic
 Able to survive the passage through the digestive system.
 Able to attach to the intestinal epithelia and colonise.
 Able to maintain good viability.
 Able to utilise the nutrients and substrates in a normal diet.
 Capable of exerting a benificial effect on the host.
 Stability of desired characteristics during processing, storage and
transportation.
 Anti-inflammatory, antimutagenic, immunostimulatory.

14. Probiotics - Mode of Action
Tiwari et al, 2012
14

15. Bacteriocins:- Bacteriocins are potent protein toxins
produced by virtually bacterial and archeal species
These bactericidal peptides play an important role in
regulating competitive interactions in natural microbial
systems
TYPES OF BACTERIOCINS
Nisin
Microcins
Colicins
pediocin

16. Advantages
Increased nutritional value
Promotion recovery of diarrhea
Inhibition of pathogen growth and translocation
Produce vitamins (especially Vitamin B and vitamin K)
Enhancing specific and nonspecific immune response
Stimulation of gastrointestinal immunity
Decreasing prevalence of allergy in susceptible
individuals

17. Guidelines for evolution of
candiadate probiotic strains

19. Isolation of probiotics :-
steps
 Collection of Samples
 Isolation
 Identification
 Acidic tolerence
 Bile tolerence
 Antibiotic senstivity

20. Isolation and identification of
Lactobacillus
 Collection of Samples:- fecal ,milk,yoghurt
samples
 Media :-MRS
 Identification:
 Gram Staining,
 catalase test

21. Tolerance of Lactobacillus casei to
gastric juice(acidic)
 One aliquat (0.2 ml) of bacteria was transfered in
a 2ml sterile eppendroff tube and mixed in 0.3ml
of 0.5% sterile Nacl solution and 1ml stimulated
gastric juice of pH 2, 3, and 4 respectively
 Viability of strains was analysed by determination
of CFU/ml after different periods of incubation.
(0min, 30min, 60min, 90min) on MRS solid
media after an incubation time of 48hrs at 37ºC

22. Tolerance of Lactobacillus casei to
gastric juice(acidic)

23. Bile tolerence
 One aliquat (0.2ml) of bacteria mixed with 0.3ml of
sterile Nacl solution and 1ml stimulated intestinal
juice (0.5%, 1% and 1.5 %) followed by incubation at
37ºC for 48hrs
 Viability of srains determined by CFU/ml after
different periods of incubation (0min, 30min, 90min,
180min) in stimulated intestinal juice by inoculation
on MRS media after an incubation time of 48 hrs
at37ºC

24. Tolerance of Lactobacillus casei to
bile juice(basic )

25. Antibiotic senstivity
 Sensitivity of Lactobacillus spp. to different
antibiotics can be determined by minimum
inhibitory concentration (MIC) or disc diffusion
assay

26. Is yogurt a probiotic product?

27. How is yogurt different from curd?
 It is a prevalent myth, what curd is to India, yogurt is
to west

28. How is yogurt different from curd?
1.Curd
( i) Obtained by coagulation of milk by harmless
lactic acid bacteria like Lactobacillus
Lactococcus lactis.
(ii) It may contain a wide variety of bacteria, which
are not defined qualitatively/quantitatively
2.Yoghurt
( i) Obtained by lactic acid fermentation of milk by
Lactobacillus bulgaricus and Streptococcus
thermophillus

29. Is yogurt a probiotic product?
 Not all yogurts contain probiotics

30. Dosage Forms….
Standard forms:
 Capsules
 Sticks
 Powder blends
 Chewable tablets

31. yakult

32.  Yakult is a probiotic dairy product made
by fermenting a mixture of skimmed milk with a
special strain of the bacterium Lactobacillus
casei Shirota.
 It was created by Japanese scientist Minoru
Shirota , who graduated from the Medical School
of Kyoto University in 1930

33. Dr. Minoru Shirota, Yakult
founder

34.  Sugar (sucrose, dextrose)
 Skimmed milk powder
 Natural flavours
 Live Lactobacillus casei Shirota strain, 6.5
billion per 65 ml bottle ( 108 CFU/mL)
 Water

36. Floraster
 While traveling in Indo china in the early 1920’s,
Henri Boulard, a French microbiologist, noticed
the locals drinking a special tea—made from
lychee and mangosteen
 S. boulardii was first introduced to the market in
1953 by the French pharmaceutical company
Biocodex. Since then, it has become available in
over 100 countries making it the worldwide

37. FLORASTOR
 250 mg Saccharomyces boulardii lyo.
 32.50 mg lactose monohydrate.
 2.85 mg magnesium stearate.
 hypromellose (outer capsule shell)
 capsule printed colouring: titanium dioxid

38. Probiotics in Skincare Products
Probiotic skincare product NUDE Skincare© was first
introduce in 2007 by NUDE Brands Ltd., UK/USA
Probiotics help balance and also stabilize microflora on the
skin
It increases certain probiotic strains in skin that protect
skin from environmental stressors, soothes skin and
improves moisture retention
38

39. Probiotic skin cream

40. Skin Probiotics Offer Best Defense
Against MRSA Infections
The research discovered this ability to prevent MRSA infection on the
skin and into wounds was due not only to the Propionibacterium acnes’
ability to inhibit MRSA biochemically: Propionibacterium acnes along
with another skin probiotic, Staphylococcus epidermidis, stimulate the
release of the body’s own immunity mechanisms through activating Toll-
like receptors
The researchers tested the ability of Propionibacterium acnes bacteria
to prevent infection on the skin of mice. The Propionibacterium acnes
bacteria cultures significantly inhibited MRSA infections – at even
greater degrees than within the laboratory. The researchers also found
that the Propionibacterium acnes bacteria tended to colonize into and
around any wounds on the skin of the mice – thereby preventing
MRSA infection into the wound.

41. 41
Johannes, 2012

42. Table The Probiotic effect of lactic acid bacteria (LAB) in human and animal health.
Medical target Example strain Reference
Prevent food allergy
Block formation of biogenic amines
Overcome lactose intolerance
Prevent diarrhea (antibiotic-induced,
rotavirus, travellers, community acquired,
Clostridium difficile colitis)
Reduce intestinal disorders and pouchitis.
Suppress side effects of
Helicobacter pylori medication with
antibiotics.
Treat Crohn’s disease, ulcerative colitis and
imflammatory bowel disease (IBD)
Stimulate anticarcinogenic activity
Treat coronary heart disease and
anticholesterolaemic effects
Control of human urinary tract infection and
vaginosis.
Immunomodulating effect
L. rhamnosus GG
L. lactis ESI 561
E. faecalis INIA 4-07
E. faecalis EFS 2
L. acidophilus
LAB
L. rhamnosus GG
L. acidophilus LB
LAB
L. rhamnosus GG
L. acidophilus
L. rhmanosus GG
B. infants UGC35624
LAB
LAB
L. acidophilus
L. acidophilus
L. rhamnosus (GG)
L. rhamnosus GR-1
L. plantarum 299
L. rhmanosus GG
Sutas et al., 1996
Joosten et al., 1996
Gilliland and Kim, 1984
Fooks et al., 1999
Heyman, 2000
Oksanen et al., 1990
Simakachorn et al., 2000
Sanders, 2003
Gionchetti et al., 2000
Kuisma et al., 2003
Canducci et al., 2000
Gupta et al., 2000
Von Wright et al., 2002
Marteau et al., 2002
Goldin, 1990
Hirayma and Rafter, 2000
Schaafsma et al., 1998
Gilliland et al., 1985
Kontiokari et al., 2001
Reid, 2001
Reid 2002
Pathmakanthan S, et al., 2004
Schultz M, et al., 2003, Passi T, 2000

43. Probiotics - Clinical Applications
 Diarrhea
 Colon cancer
 Cardiovascular diseases
 Prevention of Helicobacter pylori infection
 Allergy
Inflammatory bowel disease (IBD)
 Lactose malabsorption
 Urogenital infections
43

44. Lactose Intolerance
 Lactose intolerance is a condition in which
people have symptoms due to the decreased
ability to digest lactose, a sugar found in milk
products. Those affected vary in the amount of
lactose they can tolerate before symptoms
develop. Symptoms may include abdominal
pain, bloating, diarrhea, gas, and nausea

45. LACTOSE INTOLERNCE

46. Antibiotic Associated Diarrhea
Disease Antibiotic treatment
Disturbance of
gut microbiota
Clostridium overgrowth
produces toxins
Diarrhea46
Balance
Microbiota
Probiotics

47. Production of antimicrobial substances
Stimulation of the mucus secretion
Competition for adhesion sites
Stimulation of specific and non-specific immune responses
Probiotics
Prevention of Helicobacter
pylori infection
Helicobacter pylori Infection
47

48. Assimilation of cholesterol by bacterial cells
Deconjugation of bile acids by bacterial acid hydrolases
Cholesterol-binding to bacterial cell walls
Reduction of hepatic cholesterol synthesis
Redistribution of cholesterol from plasma to liver
Bacterial production of short-chain fatty acids
Probiotics
Reduction of blood cholesterol level
Probiotics and Heart Diseases
48

49. Reverse increased intestinal permeability
 Enhance gut-specific IgA responses
Promote gut barrier function
Modulation of immune response
Enhance IL-10 and cytokines production that promote
production of IgE antibodies
Probiotics
Beneficial in Allergy and
Atopic diseases
Probiotics and Allergy
49

50. Urogenital Tract Disorders
Production of antimicrobial substances
Competition for adhesion sites
Competitive exclusion of pathogens
Probiotics
Relief from Urogenital Infection
50

52. Probiotics and Cancer
Enzymes of Gut Flora
Glycosidase
β- glucuronidase
Nitroreductase
Probiotics
 Oligofructose plus two probiotic strains (L. acidophilus and
L. casei) supplementation in humans helped to decrease
levels of these gut flora enzymes
52

53.  LcS can augment host immune defense through induction of IL-12
production by phagocytes, in which the unique cell wall structure of
this probiotic organism plays a key role
Studies with mouse peritoneal macrophages demonstrated that
phagocytosis of LcS is essential for its induction of IL-12 production
by macrophages, but that recognition of LcS by Toll-like receptor
(TLR)2, a sensor for a variety of Gram-positive bacteria, is not
required for IL-12 induction.
The active component of LcS for the IL-12 induction has been
revealed to be the intact cell wall, and the LcS bacterial body
engulfed by the macrophages showed significant resistance to
digestion .

54. Production Of GLP-1 by probiotics
GLP-1( Glucagon-like peptide-1) is a potent antihyperglycemic hormone,
inducing the β-cells of the pancreas to release the hormone insulin in
response to rising glucose, while suppressing glucagon secretion
GLP-1 is also known to inhibit the programmed cell death (apoptosis of
pancreatic β-cells, and to stimulate β-cell proliferation and differentiation
Probiotics for the Management of
Diabetes

57. Side Effects of Probiotics
 Rare cases cause bloating, diarrhea, abdominal pain.
 If in excess cause infection that require medical attentions.
 People having on underlying disease or compromised immune
system cause potential health problems like skin rash, fever,
bloody stools etc.
 Sometimes interact with immunosupressive drugs leading to life
threating cnditions. So people taking such drugs should avoid it.

58. Adverse effects
 May be infective in severely
immunocompromised
individuals
 Introduced into a sterile body
fluids.
Probiotics effect Reference
S.Boulardii
L.Rhamnosus
Fungemia
Liver abscess
Rautio
M(1999)
Piarroux
R(1999)

59. Synbiotics
 PROBIOTICS + PREBIOTICS
 Foods containing the combination of probiotics and
prebiotics are referred to as sybiotics.
 Improved survival in upper GIT and more efficient
implantation.

60. Genetically engineered probiotics
Weight gain can be prevented with engineered probiotics,
Researchers genetically modified probiotic bacteria to produce a
hunger-suppressing compound called NAPE, which is normally
released by the cells in the small intestine after a meal and signals
the brain to reduce food intake
Studies have found that people who are obese appear to not
produce enough NAPE, which stands for N-
acylphosphatidylethanolamine, and works to suppress appetite
after having a meal

61. Institutes engaged in Probiotic research in
India
 Central Food Technology And Research Institute,
Mysore, India
 National dairy research institute, Karnal, Haryana,
India
 Institute of microbial technology, Chandigarh,
India
 National dairy development board, Anand, Gujarat,
India
 Nestle Pvt Ltd, Panipat, Haryana, India

62. Indian studies

63. Future prospects of probiotics
 The future of probiotic foods is even promising, as
modern consumers are worried to maintain their
personal health, and expect the food that they eat
to be healthy and capable of preventing illness
 These research results will probably be as
essential for the positioning of probiotic
preparations as either a food, a food supplement
or as pharmaceutical preparation

64. THANKS