Role of probiotics in Human nutrition and Healthcare

1. HUMAN PROBIOTICS AND
PREBIOTICS
INTRODUCTION
Probiotics are live microorganisms that are similar to the beneficial microorganisms
found in humans and are hence generally regarded as safe (GRAS). The concept of
probiotics (which means, “for life”) was introduced in early 20th century by Elie
Metschnikoff.
Though they have been in existence for many years, the proper appreciation of their
benefits is only a recent phenomenon. More and more research is being undertaken
to validate their efficacy; so far, such activities have demonstrated their effectiveness
in cases of antibiotic associated diarrhea, irritable bowel syndrome, lactose
intolerance, oral health etc. The functional ingredients in the probiotic products help
in balancing the intestinal microbiota resulting in enhanced over all health, well being
and boosts immune system.

2. Human intestinal microbiota.
1, mouth; 2, pharynx; 3, tongue; 4, esophagus; 5, pancreas; 6, stomach; 7, liver; 8,
transverse colon; 9, gallbladder; 10, descending colon; 11, duodenum; 12, jejunum; 13,
ascending colon; 14, sigmoid colon; 15, ileum; 16, rectum; 17, anus.
Image adapted from www.healthsystem.virginia.edu/uvahealth/adult_digest/images/
ei_0132.gif.
The gut microbiota form a diverse and dynamic ecosystem, including bacteria,
Archaea, and Eukarya that have adapted to live on the intestinal mucosal surface or
within the gut lumen.
Stomach and duodenum
• Harbor very low numbers of microorganisms: < 10^3 bacterial cells per gram
of contents
• Mainly lactobacilli and streptococci
• Acid, bile, and pancreatic secretions suppress most ingested microbes
• Phasic propulsive motor activity impedes stable colonization of the lumen
Jejunum and ileum
• Numbers of bacteria progressively increase from approximately 104
cells in the jejunum to 10^7 cells per gram of contents in the distal
ileum
Large intestine
• Heavily populated by anaerobes: 10^12 cells per gram of luminal
contents

4. HISTORY
The use of lactic acid bacteria as feed supplements goes back to pre-Christian times
when fermented milks were consumed by humans. It was not until the beginning of
this century that Metchnikoff, working at the Pasteur Institute in Paris, started to put
the subject on to a scientific basis. He believed that the flora in the lower gut was
having an adverse effect on the host and that these adverse effects could be
ameliorated by consuming soured milk. In support of this he cited the observation
that Bulgarian peasants consumed large quantities of soured milk and also lived to a
great age; he was in no doubt about the causal relationship and subsequent events
have, in part, confirmed his thesis.
He isolated what he called the 'Bulgarian bacillus' from soured milk and used this in
subsequent trials. This organism was probably what became known as Lactobacillus
bulgaricus and is now called L. delbrueckii subsp bulgaricus which is one of the
organisms used to ferment milk and produce yoghurt. After Metchnikoff's death in
1916 the centre of activity moved to the USA. The American workers showed that the
yoghurt organisms could not colonise the gut and reasoned that, if the effect was to
be manifested in the gut then a gut micro organism was more likely to produce the
required effect. Knowledge available at that time suggested the use of L. acidophilus
and many trials were carried out using this organism. Encouraging results were
obtained especially in the relief of chronic constipation.
In the late 1940's interest in the gut microflora was stimulated by two research
developments.
Firstly, the finding that antibiotics included in the feed of farm animals promoted their
growth. A desire to discover the mechanism of this effect led to increased study of
the composition of the gut microflora and the way in which it might be affecting the
host animal.
Secondly, the more ready availability of germ-free animals provided a technique for
testing the effect that the newly discovered intestinal inhabitants were having on the
host.
This increased knowledge also showed that L. acidophilus was not the only
lactobacillus in the intestine and a wide range of different organisms came to be
studied and later used in probiotic preparations.

5. PROBIOTICS
The microbial population of the digestive tract is largest in the caecum and proximal
colon with fewer numbers and species in the mouth, stomach and small intestine.
However, these smaller populations can be important for health outcomes.
The relatively sparse flora of the healthy stomach is usually less than 103 organisms
per gram of contents, comprising chiefly facultative anaerobes such as lactobacilli,
staphylococci and streptococci.
Rapid peristalsis, and the bactericidal actions of gastric secretions, limit colonisation
of the upper small intestine to usually less than 105 cells/ml of fluid. Bacterial
numbers, especially of the strict anaerobes, increase along the small intestine,
attaining a density of between 106 - 108/ml in the distal ileum.
In adult humans the bacterial population is very large, consisting of about 1014 viable
cells, which account for about 100 grams or 50% of the wet mass of colonic contents
(Cummings and Macfarlane, 1997) and of which about 15 grams of biomass is shed
daily in faeces (Cummings and MacFarlane 1991; Hill, 1995). The colonic microflora
is taxonomically diverse, consisting of over 50 genera and 400 species (Drasar,
1988; Salminen et al., 1995; Savage, 1986; Mitsuoka 1996) in which only a relatively
few genera predominate.
Probiotics are live microorganisms thought to be healthy for the host organism.
The term "probiotics" was first introduced in 1953 by Kollath (Hamilton-Miller et al.
2003). Contrasting antibiotics, probiotics were defined as microbially derived factors
that stimulate the growth of other microorganisms. In 1989 Roy Fuller suggested a
definition of probiotics which has been widely used: "A live microbial feed supplement
which beneficially affects the host animal by improving its intestinal microbial
balance".
Fuller's definition emphasizes the requirement of viability for probiotics and
introduces the aspect of a beneficial effect on the host.
According to the currently adopted definition by FAO/WHO, probiotics are: "Live
microorganisms which when administered in adequate amounts confer a health
benefit on the host".
Probiotics are commonly consumed as part of fermented foods with specially added
active live cultures; such as in yogurt, soy yogurt, or as dietary supplements.
At the start of the 20th century, probiotics were thought to beneficially affect the host
by improving its intestinal microbial balance, thus inhibiting pathogens and toxin
producing bacteria. Today, specific health effects are being investigated and
documented including alleviation of chronic intestinal inflammatory diseases,
prevention and treatment of pathogen-induced diarrhea, urogenital infections, and
atopic diseases.
The original observation of the positive role played by certain bacteria was first
introduced by Russian scientist and Nobel laureate Eli Metchnikoff, who in the
beginning of the 20th century suggested that it would be possible to modify the gut
flora and to replace harmful microbes by useful microbes. Metchnikoff produced the
notion that the aging process results from the activity of putrefactive (proteolytic)
microbes producing toxic substances in the large bowel. Proteolytic bacteria such as
clostridia, which are part of the normal gut flora, produce toxic substances including
phenols, indols and ammonia from the digestion of proteins. According to
Metchnikoff these compounds were responsible for what he called "intestinal autointoxication", which caused the physical changes associated with old age.

6. A probiotic contains thousands of genes which may potentially influence the clinical
effects. Furthermore, interaction with the host, food components or endogenous
substrates or the endogenous microbiota inside the gastrointestinal lumen may
generate by-products or end-products with functional properties. There is a great
hope that new (molecular combined with biostatistical) methods including
metagenomics and metabolomics will help to dissect the complexicity. These
methods should, for example, allow the discovery of novel genes and gene products,
with novel properties which could be later used as drugs (eg, small molecules or
proteins with antimicrobial activities, immunomodulating properties or interacting with
intestinal functions).
The majority of colonic bacteria are saccharolytic, and several bacterial groups,
including Eubacterium, Bacteroides, Bifidodobacterium and Escherichia can ferment
starch (Brown et al., 1998; Wang and Gibson 1993). Clostridium butyricum and
species of Bifidobacterium in particular are effective starch utilisers in vitro (Brown et
al., 1998).
Numerous mechanisms are involved in the action of probiotics. New discoveries
which may have an impact on the understanding of the clinical effects in IBS include
the demonstration of the presence of anti-inflammatory microorganisms in the
endogenous microbiota (especially the phylum Firmicutes),
(Sokol H,Pigneur B,Watterlot L,et al. Faecalibacterium prausnitzii is an antiinflammatory commensal bacterium identified by gut microbiota analysis of Crohn
disease patients; Proc Natl Acad Sci U S A 2008;105:16731–6.)
Bär et al reported that conditioned media from the probiotic strain Escherichia coli
Nissle 1917 modulates contractility of muscle strips isolated from humans.
(Bär F, Von Koschitzky H, Roblick U, et al. Cell-free supernatants of Escherichia coli
Nissle 1917 modulate human colonic motility: evidence from an in vitro organ bath
study. Neurogastroenterol Motil 2009;21:559–66)
Four controlled studies in humans showed that B lactis DN-173-010 shortened the
colonic transit time and two trials also showed reduction of bloating in patients IBS or
functional disorders.
(Agrawal A, Houghton LA, Morris J, et al. Clinical trial: the effects of a fermented milk
product containing Bifidobacterium lactis DN-173-010 on abdominal distension and
gastrointestinal transit in irritable bowel syndrome with constipation. Aliment
Pharmacol Ther 2009;29:104–14; Guyonnet D;
Schlumberger A, Mhamdi L, et al,. Fermented milk containing Bifidobacterium lactis
DN-173 010 improves gastrointestinal well-being and digestive symptoms in women
reporting minor digestive symptoms: a randomised, double-blind, parallel, controlled
study. Br J Nutr 2009;102:1654–62)
Previous studies showed that L farciminis CIP 103136 suppresses stress-induced
hypersensitivity in response to colorectal distension and it was recently reported that
pretreatment with this probiotic was associated with a decreased expression of Fos
protein (a marker of neuronal activation) in the spinal cord of stressed female rats.
(Ait-Belgnaoui A, Eutamene H, Houdeau E, et al. Lactobacillus farciminis treatment
attenuates stress-induced overexpression of Fos protein in spinal and supraspinal
sites after colorectal distension in rats. Neurogastroenterol Motil 2009;21:477–80)
Conditioned media from L paracasei NCC2461 reduced visceral hypersensitivity
associated with antibiotic treatment in mice and normalised substance P expression
in the myenteric and submucosal plexuses.

7. (Verdú EF, , Bercik P, Verma-Gandhu M, et al. Specific probiotic therapy attenuates
antibiotic induced visceral hypersensitivity in mice. Gut 2006;55:182–90)
L acidophilus NCFM has also been reported to increase the visceral threshold in rats
(as effectively as morphine), and to induce opioid or cannabinoid receptors on HT29
cells (a property which was not shared by a variety of other bacterial strains).
(Rousseaux C, Thuru X, Gelot A, et al. Lactobacillus acidophilus modulates intestinal
pain and induces opioid and cannabinoid receptors. Nat Med 2007;13:35–7
Role of Probiotics
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To combat diseases like as Irritable Bowel Syndrome (IBS), dyspepsia,
chronic diarrhea or constipation.
To curdle milk
To detoxify
To eliminate pathogens
To exist symbiotically
To fight against certain cancers.
To help in Lactic intolerance
To help in producing short chain fatty acids (SCFAs)
To help liver and kidneys in discharging their functions
To help regulate the immune response
To improve mineral absorption
To improve Total digestible Nutrients (TDN) of the food intake.
To inhibit LDL accumulation
To maintain optimum micro flora
To reduce Triglycerides
To reduce the stress owing to high levels of Antibiotics
Activities of Probiotic Bacteria
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Anti colon cancer effects
Anti Milk allergy
Anti-diarrheal effects
Cholesterol lowering
Correction of Hypertension
Immune system modulation
Improved tolerance to milk
Intestinal health maintenance
Reduction of Lactose intolerance
Suppression of harmful intestinal microbe activities
Suppression of pathogen translocation
Vaginal/urinary tract health maintenance
QUALITY PARAMETERS OF THE PROBIOTICS
• Adhere to gut epithelial tissue
• Be able to persist, albeit for short periods, in the gastrointestinal tract
• Demonstrate non-pathogenic behavior
• Exhibit resistance to technology processes (i.e., viability and activity in delivery
vehicles)
• Have the ability to influence metabolic activities

8. • Modulate immune responses
• Prove resistant to gastric acid and bile acid.
Lactic acid-producing bacteria are common components of probiotics. They are
popular choices because of the historical belief that these bacteria are desirable
members of the intestinal microflora, arising from the fact that lactic acid bacteria
have long been used in the manufacture of dairy foods and are thus "generally
regarded as safe", and because the consequent large-scale-culture and preservation
methods for lactic acid bacteria in a viable state have already been developed by the
dairy industry.
LIST OF SOME OF THE COMMONLY USED HUMAN PROBIOTICS
WHICH ARE CONSIDERED AS GRAS
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
17.
18.
19.
20.
21.
22.
23.
24.
25.
26.
27.
28.
29.
30.
31.
32.
Bacillius mesentericus
Bacillus brevis Migula
Bacillus circulans Jordon emend Ford.
Bifidobacterium animalis
Bifidobacterium bifidum
Bifidobacterium breve
Bifidobacterium infantis
Bifidobacterium lactis
Bifidobacterium longum
Clostridium butyricum
Escherichia coli strain Nissle 1917
Faecalibacterium prausnitzii
Lactic acid Bacillus
Lactobacillus acidophilus
Lactobacillus animalis
Lactobacillus brevis
Lactobacillus bulgaricus
Lactobacillus casei,
Lactobacillus delbrueckii subsp. bulgaricus.
Lactobacillus gasseri
Lactobacillus johnsonii.
Lactobacillus paracasei
Lactobacillus plantarum
Lactobacillus reutri
Lactobacillus rhamnosus,
Rhodospirillum rubrum
Saccharomyces boulardii
Streptococcus faecium
Streptococcus faecalis.
Streptococcus lactis
Streptococcus salivarius
Streptococcus salivarius subsp. Thermophilus
Few Examples of probiotic strains in products
Strain (alternative designations)
Bifidobacterium animalis DN 173 010
Bifidobacterium animalis subsp. lactis Bb12
Bifidobacterium breve Yakult
Bifidobacterium infantis 35624
Bifidobacterium lactis HN019 (DR10)
Brand name
Activia
Chr. Hansen
Producer
Danone/Dannon
Bifiene
Align
Howaru Bifido
Yakult
Procter & Gamble
Danisco

9. Bifidobacterium longum BB536
Morinaga Milk Industry
Enterococcus LAB SF 68
Escherichia coli Nissle 1917
Lactobacillus acidophilus LA-5
Lactobacillus acidophilus NCFM
Lactobacillus casei DN-114 001
Bioflorin
Mutaflor
Chr. Hansen
Danisco
Actimel, DanActive
Lactobacillus casei CRL431
Lactobacillus casei F19
Chr. Hansen
Cultura
Cerbios-Pharma
Ardeypharm
Danone/Dannon
Arla Foods
Information on few global suppliers of probiotics and prebiotics
Company
BioGaia
Description
Lactobacillus reuteri culture comes in three
different, producer-friendly forms: freeze-dried
powder, freeze- dried DVS (Direct Vat Set)
granules, and frozen pellets
Producer and seller of probiotic mix including L.
acidophilus and L. casei
The “nu-trish” brand probiotic culture range
consists of Probio-Tec, Yo-Fast, and other nutrish culture blends with a well-defined viscosity
profile that ferment quickly
Producer of Enterococcus LAB SF 68
The company’s cultures division produces,
develops, and markets starter cultures, media,
coagulants, and enzymes for cheese, fresh dairy,
and other food products, and also supplies
probiotic cultures for foods and supplements, as
well as natural food protectants
URL
www.biogaia.com
Producer of several brands of fermented dairy
products containing probiotics
The Lafti line of probiotics is formulated for
stability, survivability, and concentration, and
includes L. acidophilus (Lafti L10), L. casei (Lafti
L26), and Bifidobacterium (Lafti B94)
www.danone.com
GTC Nutrition
NutraFlora short-chain fructo-oligosaccharides
(scFOS) are a cane sugar or beet sugar–derived
natural prebiotic fiber
www.gtcnutrition.co
m
Lallemand
This Canadian supplier delivers probiotics and
biosupplements to the nutraceuticals,
functional-foods, and pharmaceuticals
industries
The Hi-Maize brand corn-based resistant starch
has multiple benefits, including acting as a
prebiotic for digestive health
BeneoSynergy1 is the unique, patented
oligofructose-enriched inulin prebiotic used in
the landmark SynCan project on synbiotics and
colon cancer
www.lallemand.com
Bio K +
Chr. Hansen
Cerbios-Pharma
Danisco
Danone
DSM
National Starch
Orafti
www.biokplus.com
www.chr-hansen.com
www.cerbios.ch
www.danisco.com
www.dsm.com
www.hi-maize.com
www.orafti.com

10. Probi
This biotech company develops and patents
probiotic strains, including L. plantarum 299v
and L. rhamnosus 271. L. plantarum 299 has not
yet been commercialized, but it is in the outlicensing phase
www.probi.com
Proctor & Gamble
“Align” is a probiotic supplement produced by
P&G. Align capsules contain Bifidobacterium
infantis 35624
Producer of Bacillus clausii strains O/C, NR, SIN,
and T, marketed in Europe, Asia, and South
America as Enterogermina
Frutafit inulin and Frutalose fructooligosaccharides (FOS) are soluble dietary fibers
with bifidogenic/prebiotic properties, suitable
for a variety of food systems to enrich fiber,
reduce calories, and replace sugars and fats
www.aligngi.com
Solvay
Producer of lactulose (Duphalac) for treatment
of constipation and hepatic encephalopathy
www.solvay.com
Valio
The Lactobacillus rhamnosus GG probiotic is the
most researched in the world and was recently
licensed to Dannon for the U.S. yogurt market.
The Gefilus family containing LGG is marketed
worldwide
VSL#3 is a mixture of eight strains with 450
billion live bacteria per packet
The company sells mixtures of probiotic strains
for different indications
www.valio.fi
Sanofi-Aventis
Sensus
VSL Pharmaceuticals
Winclove
www.sanofiaventis.com
www.sensus.us
http://www.vsl3.com
www.winclove.com
Evidence-based indications for probiotics and prebiotics
in gastroenterology
Disorder, action
Product
Recommended dose
Treatment of acute infectious
diarrhea in children
L. rhamnosus GG
1010–1011 cfu, twice daily
L. reuteri ATTC 55730
1010–1011 cfu, twice daily
L. acidophilus + B. infantis
(Infloran strains)
109 cfu each, three times
daily
S. cerevisiae (boulardii) lyo
200 mg, three times daily
Treatment of acute infectious
diarrhea in adults
Enterococcus faecium LAB SF68
108 cfu, three times daily
Prevention of antibioticassociated diarrhea in children
S. cerevisiae (boulardii) lyo
250 mg, twice daily
L. rhamnosus GG
1010 cfu, once or twice daily
B. lactis Bb12 + S. thermophilus
107 + 106 cfu/g of formula
Enterococcus faecium LAB SF68
108 cfu, twice daily
S. cerevisiae (boulardii) lyo
1 g or 3 × 1010 cfu per day
L. rhamnosus GG
1010–1011 cfu, twice daily
Prevention of antibioticassociated diarrhea in adults

11. L. casei DN-114 001in fermented
milk with L. bulgaricus + S.
thermophilus
B. clausii (Enterogermina strains)
Prevention of nosocomial
diarrhea in children
1010 cfu, twice daily
2 × 109 spores, three times
daily
5 × 1010 cfu, once daily
L. acidophilus CL1285 + L. casei
Lbc80r
L. rhamnosus GG
1010–1011 cfu, twice daily
B. lactis BB12 + S. thermophilus
108 + 107 cfu/g of formula
B. lactis BB12
109 cfu, twice daily
L. reuteri ATTC 55730
109 cfu, twice daily
L. casei DN-114 001 in fermented
milk with L. bulgaricus + S.
thermophilus
1010 cfu, twice daily
L. acidophilus + B. bifidum
(Cultech strains)
2 × 1010 cfu each, once daily
S. cerevisiae (boulardii) lyo
2 × 1010 cfu per day
Oligofructose
4 g, three times per day
L. rhamnosus GG
6 × 109 cfu, twice daily
B. clausii (Enterogermina strains)
2 × 109 spores, three times
daily
AB yogurt with unspecified
lactobacilli and bifidobacteria
5 × 109 viable bac, twice daily
S. cerevisiae (boulardii) lyo
1 g or 5 × 109 cfu per day
L. casei DN-114 001 in fermented
milk with L. bulgaricus + S.
thermophilus
1010 cfu, twice daily
Reduces symptoms associated
with lactose maldigestion
Regular yogurt with L. bulgaricus
+ S. thermophilus
Yogurt not heat-treated after
pasteurization contains
suitable cultures to improve
digestion of the lactose in the
yogurt
Alleviates some symptoms of
irritable bowel syndrome
B. infantis 35624
108 cfu, once daily
L. rhamnosus GG
6 × 109 cfu, twice daily
VSL# 3 mixture
4.5 × 1011 cfu, twice daily
Prevention of C. difficile
diarrhea in adults
Adjuvant therapy for H. pylori
eradication
L. rhamnosus GG, L. rhamnosus LC705, B. breve Bb99, and
Propionibacterium freudenreichii ssp. shermanii
1010 cfu, once daily
B. animalis DN-173 010 in fermented milk with L. bulgaricus + S.
thermophilus
1010 cfu, twice daily
Maintenance of remission of
ulcerative colitis
5 × 1010 viable bac, twice
daily
E. coli Nissle 1917

12. Prevention and maintenance
of remission in pouchitis
VSL# 3 mixture of 8 strains (1 S.
thermophilus, 4 Lactobacillus, 3
Bifidobacterium)
4.5 × 1011 cfu, twice daily
Treatment of constipation
Lactulose
20–40 g per day
Oligofructose
> 20 g per day
B. infantis, S. thermophilus, and
B. bifidum
0.35 × 109 cfu each strain,
once daily
L. acidophilus + B. infantis
(Infloran strains)
109 cfu each, twice daily
Prevention of postoperative
infections
Synbiotic 2000: 4 bacteria strains
and fibers including the prebiotic
inulin
1010 cfu + 10 g fibers, twice
daily
Treatment of hepatic
encephalopathy
Lactulose
45–90 g per day
Prevention of necrotizing
enterocolitis in preterm
infants
PREBIOTICS

13. A prebiotic is a selectively fermented ingredient that results in specific changes in
the composition and/or activity of the gastrointestinal microbiota, thus conferring
benefit(s) upon host health. (Gibson et al. 2010. Food Science and Technology
Bulletin: Functional Foods 7 (1) 1–19.)
Unlike probiotics, most prebiotics are used as food ingredients—in biscuits, cereals,
chocolate, spreads, and dairy products, for example. Commonly known prebiotics
are:
1
Oligofructose
2
Inulin
3
Galacto-oligosaccharides
4
Lactulose
5
Breast milk oligosaccharides
There is a growing list of candidate prebiotics such as polydextrose, soybean
oligosaccharides, isomaltooligosaccharides, gluco-oligosaccharides, xylooligosaccharides. palatinose, gentio-oligosaccharides and sugar alcohols (such as
lactitol, sorbitol and maltitol). However, the evidence for these, especially in humans,
is not as well advanced as it is for FOS and GOS.
6
Effects of Prebiotics
• Metabolic effects: production of short-chain fatty acids, fat metabolism, absorption
of ions (Ca, Fe, Mg)
• Enhancing host immunity (IgA production, cytokine modulation, etc.)
SPRCIFIC PRODUCTS BASED ON THE END USAGE:
Functional foods and beverages - dairy products, non Dairy beverages,
breakfast cereal, baked goods, fermented meat products, dry-food probiotics;

14. animal feed probiotics; dietary supplements - food supplements, nutritional
supplements and specialty nutrient.
Selection of microbes for probiotic-based strategies.
Applications:
Human application - regular consumption, probiotic therapy, prevention of
diseases and probiotic application for animals.

15. Probiotic bacteria have a variety of uses in the present society. Primarily, they are
used as a preventative measure against a variety of infections. In the study
performed by Frick et al, the scientists tested whether strains modulating
inflammatory reponses in vivo could prevent DSS induced acute colitis. Key results
were that they found that the probiotic bacteria increased the resistance of mice to
the infection. They also saw that the probiotic bacteria inhibited inflammatory
processes in vivo. The mouse model of oral Yersiniosis showed that the
administration of viable B. adolescentis (the probiotic bacteria in question) protected
mice from the generalization of Y. enterocolitica infection. In fact, in mice models with
the dextran sodium sulfate-induced- colitis, B. adolescentis attenuated the
development of colitis, indicating anti-inflammatory properties.
(Frick et al.)
Other uses for probiotics involve the commensal bacteria to be used as a neonatal
nutrition source. For example, research has shown that probiotic bacteria are
transferred through breast milk from mother to infant (Olivares et al.) As mentioned
previously, having proper intestinal flora is very important to gastrointestinal health.
Thus, for infants that are not breast-fed, proper supplements must be given with the
bacteria added to it, to ensure gut health.
MECHANISM OF ACTION BY PREBIOTICS AND PROBIOTICS
The intestinal microflora likely plays a critical role in inflammatory conditions in

16. the gut, and potentially probiotics could remediate such conditions through
modulation of the microflora.
Different strains of Lactobacillus have shown to have varying degrees of adherence.
Researchers tested the adhesive capacity of selected enterococci to human, canine,
and porcine intestinal music was investigated in order to select for potential probiotic
strains with good adhesive properties for human or animal use. The key results were
that a strong correlation was observed for the adhesion to human and canine
intestinal mucus and also between porcine and canine or human mucus. Two surface
proteins have been identified that are associated with association and may be
involved in adhesion of enteroccoci: the aggregation substance and enterococcal
surface proteins (Laukova et al.) Thus adhesion of these probiotics blocks other
pathogenic bacteria.
While it has been established that adherence is key to Lactobacillus resistance of
various gastrointestinal pathogens, studies have been done to examine where
exactly the lactobacillus adheres to, to provide said resistance. The key results were
that there were four sites of recovery for the probiotic bacteria: the cecum, the
transverse, the descending colon, the sigmoid colon (Morelli et al.).
While competition is a strong mechanism of resistance, it is not the only one used by
probiotics.
Evidence has been given to show that probiotic bacteria Lactobacilli exert
bacteriostatic or bactericidal effects against bacterial pathogens, namely,
Helicobacter pylori
(Cruchet et al.)
Further research with this pathogen/probiotic shows that Lactobacillus not only kills it,
but also prevents IL-8 release (Hamilton-Miller et al.) The authors speculate that the
mechanism by which Lactobacillus blocks Helicobacter pylori is combination of the
following: competition for nutrients, competition for exclusion, production of inhibitory
compounds, and immunomodulatory stimuli (Hamilton-Miller et al.).
Another species of bacteria that is affected by probiotics is N. gonorrhoeae. Studies
show that when probiotics are given to individuals who are infected with N.
gonorrhoeae, symptoms are alleviated. The mechanism by which this is achieved, is
again, when the Lactobacillus administered binds to the cells and therefore
preventing the adhesion of the pathogenic bacteria.
(http://www.wpi.edu/Pubs/E-project/Available/E-project-031809153710/unrestricted/finalpaper.pdf)
The direct effect of probiotics on infants’ intestinal systems by measuring changes in
systemic levels of cytokines and inflammatory markers has also been evaluated. The
researchers found that there was a significant increase in the levels of CRP (Creactive protein) in infants treated with LGG, who had AEDS. CRP is an indication of
inflammation. IL-6 induces gene activation of CRP in hepatocytes and it stimulates
CRP secretion
(Viljanen et al.)
Post inoculation with probiotic bacteria, the protein composition of the host organism
also changes. A study was done to evaluate exactly what and how changes, and the
result this causes on the organism. studies were performed in-vivo by inoculation of
Lactobacillus fermentum into the rabbit jejunum for 4 hours. After inoculation, the
bacteria were analyzed for proteome. The scientists performing this study found
changes in protein biomarkers that are beneficial for Lactobacillus and intestinal

17. epithelial cells in response to interactions. The key results were that there were
undetectable values for Lactate dehydrogenase when L. fermentum is incubated
inside rabbit jejunum. This indicates that lactate utilization is limited as a fuel in
intestinal bacterium. There is also a decrease in dihydrolipoamide dehydrogenase,
which is a component of the pyruvate dehydrogenase complex. This suggests that
oxidation of pyruvate via the krebs cycle is low post incubation. And since substrate
oxidation is associated with the production of oxygen free radicals, which are
potentially toxic to bacteria, these changes in energy metabolism would favor the
survival of L. fermentum in the intestine. Lastly, there is an increase in Glycoside
hydrolase, which plays a role in mucin degradation on the surface of the intestine.
The release of glycoside hydrolase from the bacterium may participate in the
regulation of mucin turnover and thus integrity of the intestinal epithelium. Another
important finding from this study is that the levels of some regulatory proteins were
elevated in response to incubation with the probiotic, and some of these proteins
would help protect against the inflammatory injury that occurs in response to
bacteria. They summarized that proteins can serve as biomarkers for the metabolic
changes that are beneficial for the lactobacillus and intestinal epithelial cells
(Yang et al.)
Administration of lactobacilli and bifidobacteria could theoretically modify the flora
leading to decreased β-glucuronidase and carcinogen levels (Hosada et al., 1996).
Furthermore, there is some evidence that cancer recurrences at other sites, such as
the urinary bladder can be reduced by intestinal instillation of probiotics including L.
casei Shirota (Aso et al., 1995). In vitro studies with L. rhamnosus GG and
bifidobacteria and an in vivo study using L. rhamnosus strains GG and LC-705 as
well as Propionibacterium sp. showed a decrease in availability of carcinogenic
aflatoxin in the lumen (El-Nezami et al., 2000; Oatley et al., 2000).
(http://www.who.int/foodsafety/publications/fs_management/en/probiotics.pdf)
Intravenous, intraperitoneal and intrapleural injection of L. casei Shirota into mice
significantly increased NK activity of mesenteric node cells but not of Peyer's patch
cells or of spleen cells (Matsuzaki and Chin, 2000), supporting the concept that some
probiotic strains can enhance the innate immune response.
(Joint FAO/WHO Expert Consultation on Evaluation of Health and Nutritional
Properties of Probiotics in Food Including Powder Milk with Live Lactic Acid Bacteria,
October 2001)
In a series of randomized, double blind, placebo controlled clinical trials, it was
demonstrated that dietary consumption of B.lactis HN019 and L. rhamnosus HN001
resulted in measurable enhancement of immune parameters in the elderly
(Arunachalam et al., 2000; Gill et al., 2001; Sheih et al., 2001).
Some probiotic strains were shown to inhibit the growth of enteropathogens, such as
Salmonella enteritidis, enterotoxigenic Escherichia coli, and Serratia marcesens, in
vitro and in this respect may offer considerable therapeutic potential. This finding,
together with more recent evidence showing that Lactobacillus GG exerts antagonist
activity against Salmonella typhimurium C5 infection both in vitro and in vivo (50),
provides a basis for the clinical use of probiotics in suppression of pathogens.
(http://www.ajcn.org/cgi/content/full/73/2/476S)
Potential for probiotics microorganisms to modulate the immune response and
prevent onset of allergic diseases has been demonstrated.

18. Ability of lactobacilli to reverse increased intestinal permeability, enhance gut-specific
IgA responses, promote gut barrier function through restoration of normal microbes,
and enhance transforming growth factor beta and interleukin 10 production as well as
cytokines that promote production of IgE antibodies
(Kalliomaki et al., 2001; Isolauri, 2001).
Certain microorganisms can contribute to the generation of counter-regulatory Thelper cell immune responses, indicating that use of specific probiotic
microorganisms could redirect the polarized immunological memory to a healthy one
(McCracken and Lorenz, 2001).
There is preliminary evidence that use of probiotic lactobacilli and metabolic byproducts
potentially confer benefits to the heart, including prevention and therapy of various
ischemic heart syndromes (Oxman et al., 2001) and lowering serum cholesterol
(De Roos and Katan, 2000).
There is some clinical evidence to suggest that oral and vaginal administration of
lactobacilli can eradicate asymptomatic (Reid et al., 2001a; 2001b) and symptomatic
Bacterial vaginosis
(Hilton et al., 1995; Sieber and Dietz, 1998).
One study of day care centres in Finland showed that probiotic use reduced the
incidence of respiratory infections and days absent due to ill health
(Hatakka et al., 2001).
Probiotic bacteria containing β-galactosidase can be added to food to improve
lactose
maldigestion
(Kim and Gilliland, 1983).
Studies have also shown that some infections are more prevalent in organisms and
people without a certain baseline level of probiotic bacteria. In addition to this,
research has demonstrated that having the probiotic bacteria in one’s system before
being infected leads to better recovery. For example, Helicobacter pylori infections
are seen more heavily in people that we know already have a lower baseline level of
Lactobacillus johnsonii La1. The clinical studies that were conducted show that the
Lactobacillus family of bacteria has positive effects on patients with Helicobacter
pylori infections. Results show that when the mouse models that were being tested
were fed L. salivarius prior to the challenge with H. pylori, no colonization was
achieved.
In another study, the scientists involved studied in vivo the effects of probiotics in
infants with atopic eczema dermatitis syndrome (AEDS) and those infants with cow’s
milk allergy (CMA).
They had a total of 230 in vivo subjects and found that in atopic children, there were
more coliforms and clostridia and fewer bifidobacteria and lactobacillus bacteria than
nonatopic children (Viljanen et al.). Studies were conducted on the specific effect of
probiotic bacteria on the specific effect of probiotic bacteria on the post treatment
inflammation marker, C-reactive protein (CRP). The data presented in this particular
study shows that there is a significant increase in the CRP levels in the Lactobacillustreated infants with AEDS, than those infants in the control group. This is beneficial
since the production of Human CRP leads to inhibition of production of different
inflammatory cytokines and chemokines. Human CRP also enhances the production

19. of IL-10, which displays potent abilities to suppress the antigen presentation capacity
of antigen presenting cells. It is also stimulatory towards certain T cells, mast cells
and B cells (Viljanen et al.).
Studies also show that certain probiotic bacteria can protect against various fish
pathogens, Vibrio anguillarum, Aeromonas salmonicida, and Flavobacterium
psychrophilum. These are of special importance to humans because these are
common pathogens that infect fish used for human consumption. It has been
deduced that L. rhamnosus ATCC 53103 and L. bulgaricus can be considered for
safe treatment in aquaculture (Nikoskelainen et al.) Adhesion, as will be discussed
later, is one of the mechanisms by which probiotics are thought to benefit the host
organism. In a particular study conducted by Nikoskelainen and his colleagues, it
was deduced that L. rhamnosus and L. bulgaricus were the most adhesive to the
gastrointestinal tract. This ability is thought to be the way by which probiotics
suppress pathogen growth.

20. PROOF OF ACTIVITY
Scientists have come up with techniques to follow and track the progress of the
probiotic bacteria through the gastrointestinal tract. In a study mentioned previously,
a tests were performed in vivo and the scientists stained the bacteria with cFDA-SE
and under observation, realized that the bacteria was localized in the rectum and
also in the jejunum and ileum. This is important since they know that the probiotic
bacteria can localize and adhere to the GI tract, and thus provide competition to the
pathogenic bacteria
(Bouzaine et al.)
Another method of tracking the progress of the probiotic through the GI tract is
through GFP staining. Scientists realized that bacteria can be difficult to follow
through the gastrointestinal tract, so they decided to label it with a plasmid containing
GFP. They used rats for the in-vivo models, and then took samples from the
stomach, jejunum, ileum, caecum, colon and rectum to test for presence of EcN.
They found that the EcN-GFP was detectable in high numbers in the stomach and in
lower numbers in the caecum but not in the rectum at the beginning of the
experiment. They also found out that the organisms were localized close to the
epithelial surface of the stomach, cecum, and rectum and also in the lumen of the
intestine. The transformation of EcN to obtain EcN-GFP in this study had no
detectable influence on the probiotic microorganism regarding adhesion on and
induction of IL-8 secretion of HT-29 cells and allowed the detection in mixed
microbial environments
(Schultz et al.)

21. SAFETY
Probiotics as treatment are still a novel concept in the medical world, and leads to
questions and doubts in the minds of many. However, one can rest assured that
studies are heavily underway to test for the safety and reliability of probiotics.
Research has been done to determine any toxicity present through the usage of
probiotic bacteria, Bacillus (subtilis and indicus). Researchers tested guinea pigs and
rabbits since they are considered the most sensitive lab animals. No noticeable
differences could be seen in the animals tested, thus they can assume that the
probiotic bacteria are harmless
(Hong et al.).

22. MICROENCAPSULATION_ A NOVELTY
In order to provide the consumers with the health benefits, probiotic food must
contain probiotic bacteria in sufficient number and with high viability when reaching
the human intestine, where they deliver the beneficial effect. The problem lies in the
sensibility of these bacteria to stress and extreme physiological conditions, thus the
survival of probiotics in current food is low and results in less than the recommended
effective daily intake as well as insufficient number of bacteria reaching consumers
intestine.
It is, therefore, necessary to protect probiotic bacteria added in food products until
they reach
the human large intestine. Providing probiotic living cells with a physical barrier
against adverse
environmental conditions (via microencapsulation, for instance) receives currently
considerable
interest, and quite a few research works have been conducted on the topic.
Based on a novel technology, it is aimed at devising a innovative microencapsulation
process to protect bacteria in food systems achieving at the same time a particle size less
than 30 μm (under the consumer sensory perception). The microcapsules formulations
were designed to effectively protect the bacteria by an adequate polymer coating thus
enabling them to reach the human intestine with the necessary viability level.

23. MARKET SURVEY
Beneficial microorganisms can be used in food, feed, drinking water; over pond water
mediums; in the environment.
Beneficial microorganisms can be used in controlling
• Bacterial, fungal and viral infections
• Cancer
• Cholesterol
• External parasites
• Insects
• Internal parasites
• Obesity
• Pests
Beneficial microorganisms can be used as alternate antibiotics, pesticides,
insecticides, growth promoters.
Beneficial microorganisms can be used as
• Anifungals
• Antibiotic s
• Antivirals
• Enzyme producers
• FCR Improvers
• Growth promoters
• Gut acidifiers
• Immuno modulators
• Insecticides
• Meat tenderizers
• Oxygen liberators
• Parasiticides
• Pesticides
• Pollutant degraders
• Protozoacides
• Toxin binders
• Zoothamnicides
Role of Probiotics
• To combat diseases like as Irritable Bowel Syndrome (IBS), dyspepsia,
chronic diarrhea or constipation.
• To curdle milk
• To detoxify
• To eliminate pathogens
• To exist symbiotically
• To fight against certain cancers.
• To help in Lactic intolerance
• To help in producing short chain fatty acids (SCFAs)
• To help liver and kidneys in discharging their functions

24. •
•
•
•
•
•
•
To help regulate the immune response
To improve mineral absorption
To improve Total digestible Nutrients (TDN) of the food intake.
To inhibit LDL accumulation
To maintain optimum micro flora
To reduce Triglycerides
To reduce the stress owing to high levels of Antibiotics
Activities of Probiotic Bacteria
• Anti colon cancer effects
• Anti Milk allergy
• Anti-diarrheal effects
• Cholesterol lowering
• Correction of Hypertension
• Immune system modulation
• Improved tolerance to milk
• Intestinal health maintenance
• Reduction of Lactose intolerance
• Suppression of harmful intestinal microbe activities
• Suppression of pathogen translocation
• Vaginal/urinary tract health maintenance
FACTORS THAT MAKE USAGE OF PROBIOTICS
AS THE ONLY BEST ALTERNATIVE
•
•
•
•
•
Abuse of Antibiotics
Abuse of Growth promoting antibiotics and chemicals
Abuse of Hormones
Abuse of Pesticides, insecticides
Abuse of dewormers, parasiticides, disinfectants, sanitizers etc
Probiotics, belonging to the functional group of gut flora stabilisers within the
category of zootechnical feed additives (according to the Regulation EC No
1831/2003) is a fast growing market
The probiotics market has been one of the prime beneficiaries
of the recent fad over functional foods. Rising levels of health
consciousness and the ageing baby boomer population are a few of the drivers
helping in the growth of the market. The other major market factor driving
the overall probiotics market in the present and expected to continue to do
so in the future is the influence exerted by the women buyer segment. It is
generally observed throughout the globe that traditionally women are
responsible for the buying decisions of the foods and beverages (F&B)
category in families. Since they tend to be more aware about the new products
and their health benefits, they try and incorporate more beneficial foods in
the families' diet. Hence it is no surprise that women have more knowledge
about probiotic F&B and consequently not only drive their consumption, but
also act as the opinion makers. This gathers more importance considering the
fact that the probiotic Foods & Beverages segment is expected to command over
75% of the overall probiotics market in 2009.

25. Probiotic dairy products are expected to command the highest
market share among all the probiotic foodstuffs, accounting for almost 70% in
the year 2009 and reaching a market size of almost $24 billion by the end of
2014. The biggest markets for these products are Europe and Asia; the U.S.
market has slowly but surely opened up to these products in the recent past
and is expected to grow at a CAGR of 17% from 2009 to 2014, the biggest
contributor being probiotic cultured drinks followed by probiotic yogurts.
Though the market base of probiotic products is comparatively lesser in the
U.S., the market is expected to grow at an astounding rate of almost 14% in
the same period driven by the large scale acceptance of - the probiotic
yogurts in spoonable single serve packs, probiotic cultured drinks in single
shot packaging form and probiotic dietary supplements.
Products that fall under the niche category presently, such as
probiotic chocolates, probiotic ice creams and probiotic baked products are
expected to enjoy a much larger market share due to the belief that consumers
are ready to pay an extra amount for fortified products if confident of
'extra' benefits. However, probiotic cheese, probiotic butter etc are fated
to the status of ultra-niche products due to their conventional image as
unhealthy dietary products.
The global market estimate of functional foods has been up to 73 Billion € and an
annular growth rate of 8-16%. In a recent study undertaken by Leatherhead Food RA,
the market for functional foods in the United Kingdom, France, Germany, Spain,
Belgium, Netherlands, Denmark, Finland, and Sweden was reviewed. The results of
the study showed that the probiotic yogurt market in these 9 countries totalled >250
million kg in 1997, with France representing the largest market, having sales of 90
million kg, valued at US$219 million. The German market for probiotic yogurts is
growing rapidly; for example, during 1996–1997, it increased by 150%, whereas the
UK market grew by a more modest 26% during the same period. On average,
probiotic yogurts accounted for 10% of all yogurts sold in the 9 countries studied,
with Denmark having the highest proportion (20%) of probiotic yogurts, followed by
Germany and the United Kingdom (both at 13%) and then France (11%). On the
lower end of the scale were the Netherlands and Belgium (both at 6%) and then
Finland and Sweden (both at 5%). Seen as crucial to market expansion in Europe is
further clarity on the use of health claims. The market for functional foods in Europe
could ultimately account for 5% of total food expenditure in Europe, which, based
on current prices, would equate to US$30 billion.
In 2004, the global market value of probiotics was €32 million, with a forecasted
annual growth of approximately 3%. However, due to the ban of antimicrobial feed
additives, the probiotic market in Western Europe showed an annual growth of more
than 7%. In 2006, Western Europe produced around 296 tons of probiotics, with a
value of €15.5 million. With 1012 CFU (equivalent to about 100g) usually added to a
ton of mixed feed, approximately 3 million tons of feed containing probiotics was
produced last year.
(www.allaboutfeed.net/article-database/potenti...)
In Japan a standard was developed by the Fermented Milks and Lactic Acid Bacteria
Beverages Association stipulating that a product contain 1 x 107 viable
bifidobacteria/g or mL product to be considered a probiotic food.

26. Probiotic Foods & Beverages segment is expected to command over 75% of the
overall probiotics market in 2009.
Probiotic dairy products are accounting for almost 70% in the year 2009 and reaching
a market size of almost $24 billion by the end of 2014.
Probiotic dairy products market in USA is expected to grow at a CAGR of 17% from
2009 to 2014.
Probiotic chocolates, probiotic ice creams and probiotic baked products are
expected to enjoy a much larger market share.
According to a new market research report, 'Probiotics Market (2009-2014)'
(www.marketsandmarkets.com/Market-Reports/probiotic-market-advanced-tec
hnologies-and-global-market-69.html), published by Markets and Markets
(www.marketsandmarkets.com), the global probiotics market is expected
to be worth US$ 32.6 billion by 2014, with the Europe and Asia accounting for
nearly 42% and 30% of the total revenues respectively. The global market is
expected to record a CAGR of 12.6% from 2009 to 2014.
Europe forms the largest market for probiotics with an estimated $13.5 billion by
2014. Its 12.2% CAGR from 2009 to 2014 is driven by consumer demand for healthenhancing probiotic products, such as probiotic yogurts, other probiotic dairy
products and probiotic dietary supplements.
Asia is the second largest segment, growing at with an estimated CAGR of
11.2% to reach $9.0 billion by 2014.
India is also fast emerging as a potential market for probiotics in food. Probiotic
product industry in India was estimated to be around Rs 20.6 million with a projected
annual growth rate of 22.6% until 2015.

27. STATUTORY REGULATIONS:
One major attribute of probiotics is the lack of adverse events associated with their
use.
(Gregor Reid; Canadian Research and Development Centre for Probiotics, Lawson
Health Research Institute, and Departments of Microbiology & Immunology, and
Surgery, University of Western Ontario, London, Canada; Current Pharmaceutical
Design, 2005, 11, 11-16)
International guidelines on probiotics in food broadly specify the kind of tests that
may be required to determine the safety and to assess the health claim of a probiotic
product in food. Such tests are based on the current understanding of the subject.
The regulatory mechanism for probiotics differs from country to country and also
even within a country.
In India there are no regulatory guidelines for probiotic foods upto 2011.
However a Task Force was constituted by ICMR in 2011 and the
guidelines proclaimed by it are given below.
2. GUIDELINES AND REQUIREMENTS FOR PROBIOTIC PRODUCTS
2.1. Scope: The guidelines deal with the use of probiotics in food and provide
requirements for assessment of safety and efficacy of the probiotic strain and health
claims and labeling of products with probiotics.
Note: These guidelines are not meant for probiotics which by definition would come
under drugs, beneficial microorganisms not used in foods or genetically modified
microorganisms (GMOs).
2.2 Definition of Probiotics: Probiotics are ‘live microorganisms which when
administered in adequate amounts confer a health benefit on the host’ (FAO/WHO,
2002).
2.3 Genus, species and strain identification: Effects of probiotics are strain
specific. Strain identity is important to link a strain to a specific health effect as well
as to enable accurate surveillance and epidemiological studies. Both phenotypic and
genotypic tests should be done using validated standard methodology. Nomenclature
of the bacteria must conform to the current, scientifically recognized names as per
the International Committee on Systematics of Prokaryotes (ICPS) (available at
http://www.the-icsp.org/).
The current molecular techniques used for identification include PCR based
techniques, 16S rRNA sequencing and DNA finger printing techniques like
Ribotyping and Pulsed Field Gel Electrophoresis (PFGE).
It is recommended that probiotic strains in use in India should be deposited in an
internationally recognized culture collection/repositories.
2.4 In vitro tests to screen potential probiotic strains: The following in vitro tests*
with standard methodology are recommended for screening putative probiotic strains:
2.4.1 Resistance to gastric acidity.
2.4.2 Bile acid resistance.

28. 2.4.3 Antimicrobial activity against potentially pathogenic bacteria (acid and
bacteriocin production).
2.4.4 Ability to reduce pathogen adhesion to surfaces
2.4.5 Bile salt hydrolase activity
These tests are based on the hostile gut environment which they mimic under in vitro
conditions. The cultures evaluated as probiotics based on these tests should be
subjected to preclinical validation in appropriate animal models before clinical trials
are conducted in human subjects.
2.5 In vivo safety studies in animal models: Assessment of the acute, subacute
and chronic toxicity of ingestion of extremely large amounts of probiotics should be
carried out for all potential strains. Such assessment may not be necessary for
strains with established documented use.
2.6 In vivo efficacy studies in animal models: To substantiate in vitro effects,
appropriate, validated animal models must be used first, prior to human trials.
2.7 Evaluation of safety of probiotics for human use: In recognition of the
importance of assuring safety, even among group of bacteria that are Generally
Recognized as Safe (GRAS)**, probiotics strains needs to be characterized at a
minimum with the following tests:
2.7.1 Determination of antibiotic resistance patterns. It should be ascertained that
any given probiotic strain is not at significant risk with regard to transferable antibiotic
resistance.
2.7.2 Assessment of undesirable side-effects.
2.7.3 If the strain under evaluation belongs to a species that is a known mammalian
toxin producer or of hemolytic potential, it must be tested for toxin production and
hemolytic activity respectively.
Assessment of lack of infectivity by a probiotics strain in immunocompromised
individuals would be an added measure.
2.8 Evaluation of efficacy studies in humans: The principal outcome of efficacy
studies on probiotics should be proven with similar benefits in human trials, such as
statistically and clinically significant improvement in condition, symptoms, signs, wellbeing or quality of life, reduced risk of disease or longer time to next occurrence or
faster recovery from illness. Each of the parameter should have proven correlation
with the probiotics tested.
Probiotics delivered in food may not be tested in Phase 3 studies (effectiveness),
unless the product makes a specific health claim wherein it becomes imperative to
generate the required evidence necessitating carrying out Phase 3 studies.
If a probiotic food has a record of documented long and safe use outside the country,
the data regarding this could be reviewed and deemed as sufficient to allow its
marketing within the
country. However, labeling of health benefits may require evaluation in a different
manner.
While taking into account studies done abroad, efficacy studies of probiotics (which
are of proven benefit in ‘other’ populations) should also be conducted on Indian
subjects. It is recommended that such ‘bridging’ human trials should comply with the
principles laid down by the Drug Regulatory Authority. Adverse effects, if any, should
be monitored and incidents reported to the appropriate authority.
2.9 Effective dosage of probiotic strain / strains: The minimal effective dose or
the level of viable cells of the probiotic strain in terms of cfu/ml/day in the carrier food

29. that demonstrates general health promoting functions or well being or specific health
claims in target population should be clearly indicated.
2.10 Labeling Requirements: In addition to the general labeling requirements under
the food laws, the following information should also be mentioned on the label
(23,39):
Genus, species and strain designation following the standard international •
nomenclature.
The minimum viable numbers of each probiotic strain should be specified at the level
at • which efficacy is claimed and at the end of shelf- life.
Evidence-based health claim(s) should be clearly stated. •
The suggested serving size to deliver the minimum effective quantity of the probiotic
• related to the health claim.
Proper storage conditions to be mentioned.
2.11 Manufacturing and handling procedures: Adequate quality assurance
programmes should be in place. Good Manufacturing Practices should be followed in
the manufacture of probiotic foods. The Codex General Principles of Food Hygiene
and Guidelines for Application of Hazard Analysis and Critical Control Point (HACCP)
(40) should be followed.
Guidelines for evaluation of candidate probiotic strains
Strain Identification by Phenotypic and Genotypic Methods
•
•
Genus, Species and Strain
Deposit strain in an Internationally Recognized collection
↓
Screening of Potential Probiotic Strains
• In vitroTests
↓
In vivo studies in validated animal models for:
•Safety
•Efficacy
↓
In vivo studies in humans for clinical evaluations
• Phase1 (safety)
• Phase 2 (efficacy)
• Phase 3 (effectiveness)*
↓
PROBIOTIC FOODS
↓
Labeling Requirements
• Genus, Species, Strain
•Minimum viable numbers of probiotics at the level at which efficacy is claimed and at
the end of shelf- life.
• Health claim(s)
• Serving size for efficacy
• Storage conditions
* Only required if a specific health claim is made

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