an exhaustive presentation on probiotics and prebiotics

4. What are Probiotics?
• As defined by the Food and Agriculture Organization of the
United Nations (FAO) and the World Health Organization
(WHO) in 2002, probiotics are: “Live microorganisms which
when administered in adequate amounts confer a health benefit
on the host.”
• Probiotics (literally “for life”) are friendly bacteria or yeasts and
are a concept in contrast to antibiotics. Lactobacilli and
bifidobacteria are the most common probiotic bacteria, but the
yeast Saccharomyces cerevisiae and some Escherichia coli
strains are also used as probiotics.
• Probiotics can be found in the form of food or dietary
supplements in the United States. Potential health benefits from
probiotics may vary depending on the type of probiotics
consumed.

5. RATIONALE

6. HISTORY
• At the start of the 20th century, Russian noble prize winner and father of modern
immunology, Elie Metchnikoff, a scientist at the Pasteur institute, was the first
conceptualize “probiotics”.
• In 1907 Metchnokoff proposed that the acid producing bacteria in fermented milk
products could prevent “fouling” in the large intestine and if consumed regularly,
lead to a longer, healthier life.
• In early 1930’s, in Japan, Minoru shirota developed a fermented milk product
called Yakult (probiotic yogurt like product made by fermenting a mixture of
skimmed milk with a special strain of Lactobacillus casei shirota).
• Probiotic term coined in 1965 by Lilly and Stillwell.

7. GUT MICROFLORA
Human gastrointestinal tract contains 10 times more bacteria than there are
eukaryotic cells in the body. These probiotic bacteria have:
(1) protective, (2) immunomodulatory and (3) metabolic functions.
1. Gut bacteria form a protective layer, competitively inhibit the bad bacteria,
synthesize mucus, tighten intercellular junction, thus protecting the intestinal
mucosa.
2. They help in boosting host immunity by inhibiting proinflammatory mediators,
stimulating release of antiinflammatory cytokines and releasing bacteriocins.
3. The metabolic functions are digesting carbohydrates to produce short-chain fatty
acids, synthesizing vitamin K, aiding in absorption of minerals and converting
primary bile acids to secondary bile acids

8. INTESTINAL MICROFLORA
• Rare in the esophagus
• Uncommon in the stomach
• primarily gram (+)
• 102 - 104
• 105 in the jejunum – primarily
aerobes
• 1010 – 1012 in the colon
• primarily anaerobes
• 1000x more anaerobes than
aerobes

9. FACTORS AFFECTING INTESTINAL
ECOSYSTEM
Antibiotics and other drug intake
Microbial infections
Diet (highly processed, low fiber foods)
Chronic diarrhea
Stress
Radiation and chemotherapy
Colonic therapies for detoxification

12. A SUCESSFUL PROBIOTIC MUST..
Able to survive the passage through the digestive
system. Should survive intact at low pH of
stomach.
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.
 Non pathogenic and non toxic.
Capable of exerting a beneficial effect on the host.
Stability of desired characteristics during processing,
storage and transportation.
Anti-inflammatory, antimutagenic, immunostimulatory.
In respect of the probiotic
definition of FAO/WHO
products claiming probiotic
effects should contain a
sufficient number of viable
cells to confer efficacy. This
is generally higher than
10^6–10^8 cfu/g or 10^8–
10^10 cfu/day (Champagne
et al., 2011).

13. Microorganism Strain Company (product)
Bifidobacterium adolescentis ATCC 15703
Bifidobacterium animalis Bb-12 Chr. Hansen
Bifidobacterium bifidum Bb-11 Chr. Hansen
Bifidobacterium essencis Danone® (Activia)
Bifidobacterium infantis Shirota Immunitas® Yakult Danone®
Bifidobacterium lactis Bb-02, LaftiTM B94 DSM
Bifidobacterium CRL 431
Bifidobacterium longum BB536
SBT-2928
UCC 35624
Morinaga Milk Industry
Snow Brand Milk Products
UCCork
Bacillus lactis DR10 Danisco (Howaru™)
Lactobacillus acidophilus LA-1/LA-5 Chr. Hansen
Lactobacillus bulgaricus Lb12
STRAINS

15. 1. Culture activation
in Lab
2. Preparing and
measuring materials
3. Media
preparation,
sterilization in
fermentation vessel
4. Inoculation
5. Fermentation6. Ultrafiltration
7. Loading to Freeze
dryer or Spray dryer
8. Collection of dry
powder
9. Grinding, mixing,
weighing, packaging
10. Storage at -18°C
Innoculation in
suitable matrix
PRODUCTION OF PROBIOTICS

16. FROZEN OR DRIED PREBIOTIC CONCENTRATRES
Probiotics are delivered as food supplements, generally in caplets or capsules
containing dried cells, or in processed foods.
In general, probiotic concentrates are stored until incorporation in the final products.
Microencapsulation is one of the best storage technique used for probiotics.
Microencapsulation can be done either by Gel entrapment (in alginates) or by Spray
coating.

21. Nutrient Synthesis and Bioavailability
1. Fermentation of food with lactic acid bacteria has been shown to increase
folic acid content of yogurt, bifidus milk and kefir and to increase niacin
and riboflavin levels in yogurt, vitamin B12in cottage cheese and vitamin
B6 in Cheddar cheese.
2. In addition to nutrient synthesis, probiotics may improve the digestibility
of some dietary nutrients such as protein and fat.
3. Short-chain fatty acids such as lactic acid, propionic acid and butyric acid
produced by lactic acid bacteria may help maintain an appropriate pH
and protect against pathological changes in the colonic mucosa.

22. DRUG INTERACTIONS
Since probiotics contain live microorganisms, concurrent administration of
antibiotics could kill a large number of the organisms, reducing the efficacy
of the Lactobacillus andBifidobacterium species.
Patients should be instructed to separate administration of antibiotics from
these bacteria-derived probiotics by at least two hours.
 Similarly, S. boulardii might interact with antifungals, reducing the
efficacy of this probiotic.
 Probiotics should also be used cautiously in patients taking
immunosuppressants, such as cyclosporine, tacrolimus, azathioprine,
and chemotherapeutic agents, since probiotics could cause an
infection or pathogenic colonization in immunocompromised patients.

23. SAFETY AND ADVERSE EFFECTS
When ingested orally, probiotics are generally considered safe and well tolerated.
The most common adverse effects include bloating and flatulence; however,
these are typically mild and subside with continued use.
 Constipation and increased thirst have also rarely been associated with S.
boulardii.
 One theoretical concern associated with probiotics includes the potential for these
viable organisms to move from the gastointestinal tract and cause systemic
infections. Although rare, probiotic-related bacteremia and fungemia have been
reported.
 It is estimated that the risk of developing bacteremia from ingested lactobacilli
probiotics is less than 1 per 1 million users, and the risk of developing fungemia
from S. boulardii is estimated at 1 per 5.6 million users.
 Another theoretical risk associated with probiotics involves the possible transfer of
antibiotic resistance from probiotic strains to pathogenic bacteria; however,
this has not yet been observed.

24. SIDE EFFECTS
• Since probiotics contain live microorganisms, there is a slight chance that
these preparations might cause pathological infection, particularly in
critically ill or severely immunocompromised patients.
• Probiotic strains of Lactobacillus have also been reported to cause
bacteremia in patients with short-bowel syndrome, possibly due to
altered gut integrity.
• Caution is also warranted in patients with central venous catheters, since
contamination leading to fungemia has been reported when
Saccharomyces capsules were opened and administered at the bedside.
• S. boulardii is contraindicated in patients with a yeast allergy.
• No contraindications are listed for bifidobacteria, since most species are
considered nonpathogenic and nontoxigenic.

25. DOSAGE
• Minimum Consumption: 100g of a probiotic food with
107 cfu/ g.
• most probiotics do not permanently adhere in the
intestine, but exert their effects as they metabolize and
grow during their passage through the intestine
(colonization). Thus, daily consumption of these bacteria
is probably the best way to maintain their effectiveness

26. STABILITY

27. ACID TOLERANCE
In vitro tests of acid tolerance (A) can be predictive of
survival of probiotic strains in yogurt (B). In A, the viable
counts of 3 strains of bifidobacteria were measured at time
0 (□) and after incubation for 105 min at 37°C in 0.2 mol HCl-
KCl/L buffer, pH 2.0, plus 0.1% peptone (▪)
BILE TOLERANCE
The effect of bile on the growth of Lactobacillus amylovorus
CSCC 5442 (A) and Lactobacillus amylovorus CSCC 5197 (B).
•, control (MRS broth); □, treatment (MRS broth containing
0.3% ox bile).
ADHESION TO INTESTINAL MUCOSA
Adhesion of commercial probiotic strains in 2 in vitro models
of the intestinal mucosa. □, adhesion to a differentiated
Caco-2 cell monolayer; ▪, adhesion to intestinal mucins.

28. PROBLEMS
• Probiotics are regulated as dietary supplements and not subjected to the same
rigorous standards as medications.
• A challenge with these products involves the complexity of quality control with live
microorganisms.
• As a result, individuals may obtain a product that is ineffective or that contains
varying quantities of bacteria or yeast. Published studies involving probiotics have
often utilized small sample sizes and lacked appropriate randomization, blinding, or
control groups.
• Therefore, the results from many probiotic studies should be interpreted cautiously
due to methodological limitations. There is also heterogeneity among studies, since
different probiotic doses, strains, treatment durations, and patient populations may
have been used.
• Future research needs to encompass more well-designed clinical trials in larger
populations and for longer durations to better evaluate the efficacy of probiotics.

29.  Probiotics can be consumed in the form
of capsules or through food.
 Cheese, butter, powdered milk for
infants, mayonnaise, meat, cereals, ice
cream, fruits, vegetables can serve as
vehicles for delivering probiotics.
 Dairy products are regarded as the ideal
carrier because dairy products buffer the
bacteria in the stomach and contain
functional ingredients that interact with
the probiotics.
 Encapsulated probiotics can be added to
non dairy beverages.
 The probiotics should remain viable
during the manufacture of these food
matrices without any loss in
characteristics
PRODUCT DEVELOPMENT

30. PRODUCTS
Yogurt
• Usually made from milk (rarely, from cream) inoculated with
Streptococcus thermophilus and either Lactobacillus
acidophilus or Lactobacillus bulgaricus.
• Turkish in origin
• Available in innumerable forms and flavors
1. Low fat chocolate yogurt
2. Drinkable fruit-flavored goat yogurt
3. Neon-colored yogurt in squeeze tubes

31. An organic, non-dairy,
rice “yogurt” with
whole grains and live,
active cultures,
including Lactobacillus
bulgaricus,
Streptococcus
thermophilus,
Lactobacillus
acidophilus and
Bifidobacterium
bifidum.
RICERA RICE YOGHURT
Attune makes
Wellness Bars in
three chocolate
varieties and three
yogurt and granola
varieties.
All products contain
“more than 5
times the live active
cultures in yogurt,
with less sugar.
ATTUNES CHOCOLATE AND
GRANOLA BARS
Vive contains
one billion
CFUs of
Lactobacillus
acidophilus per
serving.
KASHI VIVE PROBIOTIC
WELLNESS CEREAL

32. • First probiotic juice
launched in the fall of 2007
by Next Foods.
• Goodbelly, organic fruit
juice-based probiotic
beverage , contains
L.Plantarum 299v, has
effects on irritable bowel
syndrome
• Three initial flavors include
Brilliant Blueberry, Peach
Mango and Strawberry
Rosehip
PROBIOTIC JUICE

33. DOSAGE FORMS
Standard forms:
– Capsules
– Sticks
– Powder blends
– Chewable tablets
Individual customization:
– Capsules
– Sticks
– Chewable tablets
– Sachets
– Tablets

37. What are Prebiotics?
Prebiotics are non-digestible substances that when consumed provide a
beneficial physiological effect on the host by selectively stimulating the
favorable growth or activity of a limited number of indigenous
bacteria.(Gibson and Roberfroid 1995)
New Definition (By Roberfroid 2007): Prebiotics are selectively
fermented, dietary ingredients that result in specific changes in the
composition and/or activity of the gastrointestinal microbiota, thus
conferring benefits upon host health.
Prebiotics modify the balance of the intestinal microbiota by stimulating
the activity of beneficial bacteria such as Lactobacilli and Bifidobacteria

38. CLASSIFICATION OF PREBIOTICS

39. STRUCTURES

41. SOURCES
Food Prebiotic Fiber Content by Weight
Raw Chicory Root 64.6%
Raw Jerusalem Artichoke 31.5%
Raw Garlic 17.5%
Raw Leek 11.7%
Raw Onion 8.6%
Raw Asparagus 5%
Raw Wheat bran 5%
Whole Wheat flour, Cooked 4.8%
Raw Banana 1%
While there is no broad consensus on an ideal daily serving of prebiotics, recommendations
typically range from 4 to 8 grams (0.14–0.28 oz) for general digestive health support, to 15 grams
(0.53 oz) or more for those with active digestive disorders.

42. PREBIOTIC MARKET

43. Reduced Constipation.
As a soluble dietary fiber, inulin shortens fecal
transit time, slightly increases fecal bulk.
Barrier against pathogens.
The SCFAs reduce lumenal pH in the colon to
levels at which pathogens such as E. coli can’t
multiply effectively.
In addition, increased populations of
bifidobacteria and lactobacilli can compete with
other organisms for nutrients and receptors.
Cancer Prevention.
Butyrate, a product of fermentation of prebiotics
by gut microflora, promotes the apoptosis of
carcinogenic cells and is thought to be protective
against colon cancer.
Improved Calcium absorption.
SCFAs cause a reduction in lumenal colonic pH
which is likely to increase calcium solubility and
overall levels in the gut.
SCFA enter the colon in a protonated form and
then dissociate in the intracellular environment.
The liberated proton is then secreted into the
lumen in exchange for a calcium ion.
Cholestesterol Lowering effect.
The prebiotic fiber bind with bile acids and
reduce solubilisation of cholesterol leading to
a cholesterol lowering effect. This overall
cholesterol lowering effect could reduce the
stiffness of large arteries and thus could
potentially reduce blood pressure.
Insulin resisistence lowering effect.
Insulin resistance is a physiological condition
where the natural hormone insulin becomes
less effective at lowering blood sugars.
The prebiotics have been found to improve
postprandial glucose response and decreased
secretion of insulin via lowered glycemic
index.
HEALTH BENEFITS

44. HEALTH
BENEFITS
Immune
function
Colon Cancer
Microflora
Modification
Blood
Glucose
Lipid
Metabolism
Mineral
Absorption
Laxation
Diarrhoea  Most of these effects
possibly emanate from
increased production of
short-chain fatty acids by
the stimulated colonic
bacteria.
SIDE EFFECTS
Prebiotics may cause
unpleasant side effects like
gas, bloating and increased
bowel movements.

45.  Prebiotics are not digested by our
digestive system because of the beta-
configuration of the bonds in their
structure.
 Inulin, oligofructose, and FOS are fully
metabolized by the colonic microflora.
 The end products of fermentation are
gases(such as carbon dioxide and
hydrogen), lactates, and short-chain
fatty acids (like acetate, propionate,
and butyrate).
 The Degree of Polymerization(DP)
influences the metabolism of specific
inulin-type prebiotic compounds.
MOA

47. MECHANISM

49. PRODUCTION OF PREBIOTICS
(1) direct extraction of natural oligosaccharides from plants;
(2) controlled hydrolysis of natural polysaccharides; and
(3) enzymatic synthesis using hydrolases and/or glycosyl transferases from plant or
of microbial origin.
Prebiotic Method
Inulin (FOS) Hot water extraction from Chicory root
followed by enzymatic hydrolysis
GOS Enzymatic lactose Transgalactosylation
XOS Enzymatic hydrolysis of plant xylans
Isomaltooligosaccharide Transglucosylation of liquified starch
lactulose Isomerization of lactose

51. 1. Mainly fructooligosaccharides(FOS)
are produced by this method.
2. The process is called
transfructosylation.
3. The fungal enzyme beta-fructosidase,
derived from Aspergillus niger is used.
4. Starting molecule used is sucrose,
and the enzyme activity sequentially
adds fructose units with new beta
(2−1) linkages placed in the chain.
5. Unlike inulin and oligofructose,
transfructosylation does not result
exclusively in beta (2−1) fructosyl-
fructose glycosidic bonds; other
linkages occur in limited numbers.
6. DP ranges from 2-4.
ENZYMATIC PROCESS

52. Chicory Roots
Washing
Slicing
Sun Drying (30-
35°C) or Hot
Oven Drying (80-
90°C)
Grinding to get a
chicory powder
Extraction by
diffusion in hot
water (70-80°c
for 1 hr)
Filtered using a
double layer
muslin cloth
Precipitation of
filtrate using
20,40, 60%
ethanol
Inulin obtained
by centrifugation
& dried at 45°C
INULIN

53. EMERGING
PREBIOTICS
1) LACTULOSE
2) LACTOSUCROSE

54. FUNCTIONAL PROPERTIES
Food Products Applications
Dairy Products Body and mouth feel; Sugar and fat replacement; Synergy
with sweeteners; Foam stability
Frozen Desserts Melting and texture; Sugar and fat replacement; Synergy with
sweeteners
Table Spreads Emulsion stability; Fat replacement; Spreadability and texture
Baked goods and
breads
Retention of moisture; Sugar replacement
Breakfast Cereals Crispness and expansion
Fruit preparations Body and mouth feel; Sugar replacement; Synergy with
sweeteners
Meat products Texture and stability; Fat replacement
Chocolates Heat resistance; Sugar replacement

55. NUTRIENT-NUTRIENT INTERACTION
• Uptake of calcium and magnesium is crucial for bone structure and
increasing absorption can prevent conditions such as osteoporosis.
• Chonan et al. (2001) have shown that adding GOS to the diet of rats can
increase calcium and magnesium absorption.
• The mechanism for this is unclear but in this case the presence of a colonic
flora is required for GOS to have this effect, though the authors
acknowledged that microbial mediated and nonmicrobial mediated
mechanisms probably exist.
• FOS can also affect mineral absorption and in human studies 15g per day
oligofructose or 40 g per day inulin increased the apparent calcium
absorption
• Magnesium absorption has also been shown to increase when ingesting
FOS

56. NUTRIENT-NUTRIENT INTERACTION
• Prebiotics may also have an effect on lipid regulation. Although the
mechanism is currently unknown, studies have shown positive results and
mechanistic hypotheses have been developed.
• A study on diabetic rats found that when XOS replaced simple
carbohydrates in the diet, the serum cholesterol and triglyceride
increases observed in diabetes were reduced and liver triglycerides
increased to a comparable level seen in healthy rats (Imaizumi et al., 1991).
• Other studies have examined FOS, which was also found to reduce
blood lipids (Bornet et al., 2002; Roberfroid, 2002). This was thought to be
due to an inhibition of lipogenic enzymes in the liver, which may be a
result of the action of propionate produced from the fermentation of
prebiotics by gut bacteria

57. DRUG-NUTRIENT INTERACTION
• Drug interactions include a reduction in the effectiveness of the blood
thinner warfarin and the arthritis medication sulfasalazine

59. Forms and Dosage
• Isapghula husk as a loose powder. A good dose is 10 grams(= 2 heaped teaspoons) twice
daily. For IBS.
• FOS(fructooligosaccharides) loose powder. A good dose is 2.5 grams(= 1 level teaspoon)
twice daily.
• Inulin powder/ syrups 2.5 grams(= 1 level teaspoon) twice daily( better prebiotic than
FOS)
• GOS( Galactooligosaccharides) 5 grams daily
• Butyric acid capsules- Provide most of the benefits of probiotics, without actually taking
probiotics for upset of gut.
• Start with lower doses for a few days, otherwise may experience some constipation.
• Caution is taken with antibacterial herbs or supplements that are taken along with,
which may inhibit the good bacteria as well as the bad.
The wonders of prebiotic.Hip, Oct 6, 2010

60. SAFETY
Fructooligosaccharides:
• Optimum intake : 10 gm/day
• The doses upto 44gm for men and 49gm for women are supposed to
induce diarrhea
Inulin and oligofructose:
• There is no evidence of toxic effects
• The dose of intolerance is quite high, which allows for a broad therapeutic
dose range

61. PROBLEMS WITH PREBIOTICS
• Overdose can cause intestinal bloating, pain, flatulence, or diarrhea
• Not as potent as antibiotics in eliminating specific pathogens
• Overcomes the need for probiotic bacteria to compete with intestinal bacteria
that are well established in their niche

62. PROBIOTICS v/s PREBIOTICS
Stable in long shelf life foods and
beverages
Physicochemical properties useful to
food taste and texture
Resistant to acid, protease, and bile
Stimulate organisms already resident
in the host
Stimulate fermentative activity of the
microbiota and health benefits from
SCFA
Lower intestinal pH and provide
osmotic water retention in the gut
Safe for long-term consumption and
prophylactic approaches
Do not stimulate side effects/ antimicrobial
resistance genes
Not allergenic
PROBIOTICS v/s ANTIBIOTICS

64. 60 CAPSULES
Rs 600/-

66. CONCLUSION
• The healthful effects of pre- and probiotics factor in their potential impact on
the balance of the body’s microflora, and directly or indirectly in their
enhancement of the function of the gut and systemic immune system.
• Although benefits vary, depending on the type and amount of a pre- or
probiotic consumed, experts agree that daily consumption of foods
containing these functional components is beneficial.
• In addition, effects of probiotics are strain-specific and must be
demonstrated through appropriate clinical trials.