PROBIOTICS

PROBIOITCS

Prepared by
T.Krishnakumar
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S.No Contents Page.No

1. Introduction

2. Historical perspective

3. Facts that make bacteria- a probiotic

4. Probiotics foods

5. Promising health benefits of probiotics

6. Probiotic products

7. Safety of probiotics & recommended

8. Future perspectives
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INTRODUCTION

“Let food be thy medicine and medicine be the food” the old age quote by
Hippocrates, is certainly true in case of probiotics. Probiotics are live microbial food
ingredients, which have beneficial effects on human health by enhancement of immune
system. It is one of the functional foods that can positively modify the biological and
physiological processes in human nutrition. For thousands of years probiotics are used in
foods and alcoholic fermentation and in past century the scientist had find their ability to
prevent and cure the various human diseases.

PROBIOTICS

The term probiotics was derived from the Greek word “meaning for life” was
originally used by Lilly and Stillwell. In 1965 it is described
as a substances secreted by one microorganism that stimulate
the growth of another. Parker (1974) refereed probiotics as
“Organism and substances, which contribute to the intestinal
microbial balance and included microbial culture suspension
and crude microbial culture products. Gibson and Roberfoid
(1995) said that probiotics are microbial food or feed
supplements that beneficially affect the host by improving its
intestinal microbial balance.
The United States Food and Drug Administration (FDA) in 1989 used the term
“Direct Fed Microbial (DFM)” instead of probiotics. The FDA defines DFM as a source of
live naturally occurring microorganisms. In 2001, a joint committee Food and Agriculture
Organization of the United Nations/World Health Organization (FAO/WHO) redefined
probiotics as “live microorganisms that, when administered in adequate amounts, confer a
health benefit on the host.”
HISTORICAL PERSPECTIVE
The first recorded probiotic was fermented milk for human consumption. After that,
probiotic become popular with animal nutrition. The concept of the probiotics was found in
the Bible and the sacred book of Hinduism. In 76 B.C., the Roman historian Plinio suggested
the administration of fermented milk products for treating gastrointestinal infection.
But the original observation was done by the Russian scientist Metchnikoff. In 1970,
he mentioned that the consumption of foods such as yogurt, kefir and sour milk contacting
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lactic acid bacteria was associated with good health. In his book
“The Prolongation of Life” he suggested that the long, healthy
life of Bulgarian peasants resulted from their consumption of
fermenting Bacillus which positively influenced the micro flora of
the colon, decreasing the toxic microbial activities. In the
beginning of the 20th century, Tissier, proposed that Bifidobacteria
might be effective in preventing infections in infants, as they were
the predominant component of the intestinal microflora of breast-
fed infants.
In 1926, Henneberg proposed the use of an intestinal
isolate, Lactobacillus acidophilus, to produce what he called “acidophilus-milch,” or “reform
yogurt.” This concept finally became a success in the 1980s in Germany and other Western
European countries and the Lactobacillus species used for the fermented product referred to
as “yogurt mild” were selected on the basis of their technological properties, and not their
potential health benefits.
According to an anecdote told by Murch, Adolf Hitler was first reported to have
benefited from probiotic intervention: Hitler’s eczema and irritable bowel symptoms were
result from imbalance in intestinal microbes and cured by consuming the probiotic
Escherichia coli Nissle 1917.
FACTS THAT MAKE BACTERIA- A PROBIOTIC
Based on the three facts the bacteria were considered as a probiotic as follows:
1. It must be safe for consumption. As per the law of Generally Recognized As Safe
(GRAS), only safe bacteria are accepted on the market.
2. Scientific proof must be for the specific bacteria and it should produce measurable
beneficial effects on health.
3. The bacteria must be alive when reaching the human gut. It must be recommended
with the sufficient dose from the manufacture and till the time of consumption.
GENERAL PROPERTIES OF PROBIOTIC
The probiotic have several properties generally (table 1) they are:
 Safety
 Functional behavior (survival, adherence, colonization, antimicrobial production,
immune stimulation, anti-genotoxic activity and prevention of pathogens such as
Helicobacteria pylori, Salmonella, Listeria and Clostridum)
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 Technological aspects (growth in milk, sensory properties, stability, phage rsistance,

viability in processes).
Table 1: Desirable properties of probiotic microorganism

PROBIOTICS FOODS
Probiotics can be in powder form, liquid form, gel, paste, granules or available in
the form of capsules, sachets, etc. Probiotics can be bacteria, moulds, and yeast (Table
2). It is incorporated with the foods; the best known example of a probiotic food is
yogurt. Other foods are:
 Acidophilus milk
 Buttermilk

 Kefir (fermented milk-based drink)

 Tempeh (fermented soy product)

 koumis
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 Bidifus milk

 Other "fermented" foods like sour cream, cheese, sour dough bread, wine beer,
fermented meats and fermented vegetables like sauerkraut which typically do not
contain live cultures in the finished products.

There are other food components called prebiotics, which stimulate the growth of probiotics.
Prebiotics are found naturally in many fruits and vegetables as well as in whole grain foods.

 Oatmeal
 Flax

 Barley

 Other whole grains

 Onions

 Green vegetables (especially dandelion greens, but

also spinach, collard greens, chard, kale, and mustard
greens)

 Berries, bananas, and other fruit

 Legumes (lentils, kidney beans, chickpeas, navy beans, white beans, black beans,
etc.,)

Table 2: Microorganisms commonly used as probiotic around the world

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SCREENING, IDENTIFICATION AND CHARACTERIZATION OF STRAINS

Several genera of bacteria have been proposed as probiotic cultures, the most
commonly used are Lactobaillus and Bifidobacterium species. However, the selection of a
strain to be used as an effective probiotic is a complex process.

According to FAO/WHO guidelines it is necessary to identify the microorganism to

species/strain level given that the evidence suggests that the probiotics effects are strain
specific. It is recommended to employ a combination of phenotypic and genetic techniques
to accomplish the identification, classification and typing.
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For the nomenclature of bacteria, scientifically recognized names must be employed

and it is recommended to deposit the strains in an internationally recognized culture
collection. Further characterization of strains must be undertaken taking into account the
“functional” or probitic aspects and safety assessment. In vitro tests, some of them
summarized in Fig, are useful to gain knowledge of both strains and mechanism of the
probiotic effect.

In addition, even if these genera have a long history of safe consumption in

traditionally fermented products and several species have been awarded a “General
Recognised As Safe” (GRAS) status by the American Food and Drug Association or a
qualified presumption of safety (QPS) consideration by the European Food Saftey Authority
(EFSA), some characteristics must be studied to ensure the safety of the novel lactobacilli
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and bifidobacterium strains. Several of the in vitro tests can be correlated with in vivo studied
with animal models, but probiotics for human use must be validated with human studies
covering both safety (phase 1 trails) and efficacy (phase 2 trails). Phase 2 studies should be
designed as double –blind, randomized to measure the efficacy of the probiotics strain.

PROMISING HEALTH BENEFITS OF PROBIOTICS

Lifestyle and eating habits are partly responsible for each individual’s overall health
status. In ancient times, human species would have definitely eaten food with lot of live
bacteria. As the concept of hygiene improved, we began eating clean food with very little live
bacteria. The diet in western countries has been associated with the development of diseases
such as heart disease and cancer because of its content of saturated fats and sugars and
relative lack of fruits and vegetables and the desirable omega-3 fatty acids.

Figure 1: Overview of probiotics role in human

Western diet contains dramatically decreased numbers of fermented foods

coincidence that the increase in inflammatory conditions, allergic conditions, obesity, heart
disease and cancers has paralleled the decreased Probiotics. They impart numerous nutritional
and therapeutic benefits to the host. In some instance, the probiotic microbes function for the
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benefit of the host, such as antimicrobial mechanism. While in other cases, the probiotic
microbes may trigger the indigenous microbes (Figure 1).

Probiotics are widely thought to aid in restoring the balance of the intestinal flora
(Figure 2), providing protection against infection while improving metabolism. The challenge
is to ensure the survival and activity of the orally ingested microorganisms within the harsh
environment of the human digestive tract.

Figure 2: Normal flora of GI tract.

The beneficial attributes of probiotics can be broadly grouped as: antimicrobial,

biochemical, physiological and immunological.

1. ANTIMICROBIAL PROPERTIES:

It is now recognized that probiotic bacteria such as

lactobacillus and bifidobacteria have antimicrobial
properties against a variety of underirable enteric pathogenic
bactiera such as Salmonella typhimurium, E. coli, C. diffcile
and Shigeilla spp. Etc. Mechanism responsible for the
inhibition of pathogens include.
Competition for nutrients and adhesions sites
Production of inhibitory metabolites, such as organic acid, hydrogen peroxide,
bacterocins and deconjugated bile acids
Stimulation of immune system
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There are various studies indicating that consumption/ application of probiotic bacteria is
effective in the treatment of:

Probiotic-Derived Antibacterial Substances

Probiotics exert direct antibacterial effects on pathogens through production of
antibacterial substances, including bacteriocins, acid, and hydrogen peroxide.
Bacteriocin
Bacteriocins are small antimicrobial peptides produced by Lactobacilli. The primary
mechanism of bacteriocin action is forming pores in the cytoplasmic membrane of sensitive
bacteria, but they can also interfere with essential enzyme activities in some species. In
addition, several strains of Bifidobacteria have been found to produce bacteriocin-like
compounds toxic to both Gram-positive and Gram-negative bacteria. Interestingly, probiotics
also stimulate intestinal epithelial cells to produce antimicrobial substances (Table 3).
Table 3: Some bacteriocins synthesis organism

Acid
Probiotic bacteria, especially strains of Lactobacilli , produce acetic, lactic, and
propionic acid which lower the local pH leading to growth inhibition of a wide range of
Gram-negative pathogenic bacteria. Some Lactobacillus strains inhibit the growth of S.
enterica solely by the production of lactic acid. However, antibacterial effects of other
strains of Lactobacilli may be the result of a combination of lactic acid and additional
unknown Lactobacillus -derived bactericidal substances. One study suggests that lactic acid
or low pH acts as a permeabilizer of Gram-negative bacterial outer membranes, allowing
other antimicrobial substances to penetrate and increasing the susceptibility of pathogens.
Hydrogen Peroxide
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The production of hydrogen peroxide exerts a nonspecific antimicrobial defense

mechanism in the normal vaginal ecosystem. Several Lactobacillus strains of vaginal origin
have been found to produce hydrogen peroxide to inhibit gonococci growth in the female
genital tract. Additionally, inhibition of Gardnerella vaginalis by Lactobacillus strains is
through the combinational effects of hydrogen peroxide, lactic acid, and bacteriocins. General
mechanism of metabolic products are given in the Table 4.

Table 4: Action of metabolic product of probiotic on pathogens

There are various studies indicating that consumption/ application of probiotic bacteria is
effective in the treatment of:

INFANTILE DIARRHEA

The most studied gastrointestinal condition treated by probiotics is acute infantile

diarrhea. Rotavirus is the leading cause of infantile diarrhea worldwide and rapid oral
rehydration is the primary treatment. Several potential mechanisms have been proposed for
how lactobacilli reduce the duration of rotavirus diarrhea via: competitive blockage of
receptor sites in which lactobacilli bind to receptors, enhanced immune response signals from
lactobacilli bind regulates the secretary and motility defenses designed to remove perceived
noxious substances and that lactobacilli produced substances that inactivate the viral
particles.

ANTIBIOTIC ASSOCIATED DIARRHEA

The incidence of antibiotic associated diarrhea in children ranges from 20% to 40% of
those receiving broad spectrum antibiotics. The infants are more prone to get diarrhea
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because of immature intestinal mucosal barrier, immature systemic immunity as well as

incomplete evolved intestinal bacterial flora which takes 1 year to reach adult level and
composition.

The mechanism by which antibiotic, chemotherapy or

radiation associated diarrhea occurs most likely relates to disturbances
of microbial flora in gastrointestinal tract which has profound effect
on the protection barrier and results in overgrowth by pathogens,
invasion and translocation of toxins and life threatening infections,
e.g. Clostridium difficile. Antibiotic may also disturb the metabolism
of carbohydrates, resulting in malabsorption of osmotic ally active
particles.

Antibiotic-associated diarrhea and gastrointestinal problems are an important area of

probiotic application. Reported control clinical studies indicate that antibiotic-associated
diarrhea can be prevented by Lactobacillus GG strain in yoghurt form using a cfu/ml in
healthy volunteers or as a freeze-dried product using doses varying from 10 9 to 1010 cfu/day
in infants receiving antibiotics. There are also good studies showing the efficacy of a
probiotic yeast Saccharomyces boulardii in the treatment of antibiotic-associated diarrhea.

TRAVELLER’S DIARRHEA

Though a large number of microorganism are responsible for traveller’s diarrhea but
the most commonly found are enterotoxigenic E.coli. according to one report the incidence
of diarrhea in Danish tourists to Egypt was reduced to 43 percent in treated group from 72
percent in control. The probiotics given were mixture of live lyophilized L. acidophilus, B.
lactis, L. bulgaricus and S. termophilus. The mechanisms for the resistance of enteric
pathogens are:

Secretary immune effects

Colonization resistance
Alteration of intestinal conditions to be less favorable for
pathogenecity (pH, short chain fatty acids, bacteriocins)
Alteration in toxin binding sites
Influence on gut flora population
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Adherence to intestinal mucosa, interfering with pathogen adherence

Upregulation of intestinal mucin production, interfering with pathogen attachment to
intestinal epithelial cells

CHRONIC GASTRITIS
Helicobacter pylori is a microaerophilic and gram negative bacillus that causes
chronic gastritis, peptic ulcer diseases and also suspected to be involved in the genesis of
malignant gastric ulcer. The inhibitory effect of lactobacilli on the growth of H. pylori is
dependent on an extracellular secretory product, probably lactic acid which causes inhibition
of colonization.

UROGENITAL INFECTIONS
Probiotics bacteria decrease the level of uraemic toxin like phenol, pcresol and
indicant and inhibit the production of hydrogen peroxide or biosurfactatn, thereby preventing
the urogential infection. Effective colonization resistance and adhesion to urinary and gential
tract cells have also been shown to be responsible for it.

Bacterial vaginosis, yeast vaginitis and recurrent urinary tract infections (UTIs) are
common urogenital problems. The normal
vaginal flora in premenopausal women
consists primarily of lactobacilli, which are
protective against infection, but many
pathophysiologic factors can cause unstable
vaginal flora that may result in infection.
There are only few studies with the use of
probiotics in vaginal infections. The
overwhelming experiences have revealed a positive effect of L. acidophilus. Evidence from
the available studies also suggests that probiotics can be beneficial for preventing recurrent
UTIs in women. The prevention or resolution of bacterial vaginosis is particularly important
in women at risk of human immunodeficiency virus (HIV) infection. Thus treatment of
bacterial vaginosis and promotion of vaginal lactobacilli may reduce a woman's risk of
acquiring HIV-1, gonorrhea and trichomoniasis.

2. BIOCHEMICAL EFFECTS
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Micrbial fermentation tends to increase the content of various nutritents in the food.
Termed as bio-enrichment, it involuves enrichment of food value by supplementation with
proteins, essential amino acids, minerals and vitamins. For example, lactic acid bacteria
increase folic acid in yogurt, bifidus milk and kefir, niacin and riboflavin levels in yogurt,
vitamins B12 in cottage cheese, and vitamin B 6 in cheddar cheese. Probiotic microbes may
release various enzymes into the intestinal lumen, thus aiding the digestion process.

REDUCTION OF SERUM CHOLESTEROL

Large dietary intake of yoghurt lowered serum cholesterol. One of the important
beneficial effects attributed to the L. acidophilus group is its ability to reduce serum
cholesterol, possibly through binding of the dietary cholesterol with the cells or deconjugate
the bile salts in small intestine and prevent the re absorption in the liver.

Probiotics have been reported to have cholesterol lowering properties in human. The
proposed mechanism includes direct assimilations of lipids and conversion of lipids into their
metabolites. In addition, the end products of probioitcs may affect synthesis of cholesterol.
Some studies suggested that deconjucation of bile acids by probiotics results in less efficient
absorption of cholesterol from the intestine.

LACTOSE HYDROLYSIS

Lactose-intolerant individuals, because of a genetic disorder, are unable to

produce lactose (β-galactosidase) in the small intestine. When they consume milk, lactose
molecules are not hydrolyzed in or absorbed form the small
intestine but passed to the colon. They are then hydrolyzed in
the colon by lactase of different bacteria to glucose and
galactose and then further metabolized to produce acids and
gas, resulting in fluid accumulation, diarrhea, and flatulence.
Consumption of yogurt, acidiophilus milk, and live cells of
lactobacillus, especially Lab. acidophilus in fresh milk and
pharmaceutical products, reduces the symptoms in lactose-intolerant individuals. This
benefit is attributed to the ability of beneficial bacteria to supply the needed lactase in the
small intestine. However, as Lab. delbrueckii ssp. Bulgaricus and Str. Thermophilus do not
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survive in stomach acidity well and are not normal intestinal bacteria, the benefit of
consuming normal yogurt is considered to be due to the reduced amounts of lactose in yogurt,
as compare to milk, and to supply of lactase from dead cells . In contrast, the intestinal
bacteria, especially some lactobacillus species, could under proper conditions, colonize the
small intestine and subsequently supply lactase.

ANTIMUTAGENIC AND ANITCARCINOGENIC PROPERTIES

Colorectal cancer is one of the leading causes of mortality in both men and
women. In several countries, a positive correlation between dietary factors, such as
consumption of red meat and fat and the incidence of colon cancer has been established.
Probiotics especially lactic acid bacteria have been found to be effective in
prevention and suppression of cancer of the colon. At the present, several studies have been
reported on cancer and intestinal microecology related aspects. Some of the mechanisms by
which large-bowel cancer may be prevented are:
 Metabolic conversion/degradation/absorption of carcinogenic compounds
 Reduction in levels of faecal bacterial enzymes
 Stimulation of immune system.
The reports on the benefits of oral administration of probiotic culture milks and lactic acid
bacteria on tumors have been connected with changes related to tumor induction and
promotion. The following mechanism has been indicated in various studies relating lactic
acid bacteria intake:
 Altered conversion of precarcinogens to carcinogens
 Decrease or delay in the absorption and bioavailability of harmful components and
toxins.
 Enhanced resistance to inflammation, chemicals and other factors
 Strengthened intestinal barrier mechanisms

Lactic acid bacteria have been reported to inactivate dietary and intestinally
generated mutagenic compounds such as azodyes and n-nitrosamines. It has been proposed
that peptidoglycan and polysaccharide
components of bacterial cell walls have strong
binding affinities for mutagen. Lactobacillus
acidophilus, Lactobacillus casei Shirota strain
and Lactobacillus GG have been shown to have
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inhibitory properties on chemically induced tumors in animals. The proposed mechanisms

included the enteropathogens such as E. coli and C. perfringens produce enzyme β-
gulcosidase and urease, these enzymes can convert procarcinogens to proximate carcinogens.
Probiotics by lowering the intestinal pH create a bacteriocidal environment and thus may
modulate these bacterial enzymes. Probiotics colonizing at the expense of enteropathogens
may bind the ultimate carcinogen by physically removing it via feces. The cell wall of B.
infants induces activation of phagocytes to destroy growing tumor cells.
3. PHYSIOLOGICAL MECHANISMS
IMMUNE RESPONSE
i) THE ROLE OF INNATE IMMUNITY IN PROBIOTIC ACTION
Innate immunity serves to recognize and react to microorganisms and to either destroy
them if possible or contain them if they cannot be eliminated. The machinery of innate
immunity includes soluble factors like defensins and lysozyme as well as cells like
macrophages, Poly-Morphonuclear Neutrophils (PMNs), mast cells and dendritic cells.
Intestinal epithelial cells which can produce a variety of immunomodulatory proteins and
cytokines in response to microbes can also be considered part of the innate immune system in
the gut. In theory, probiotics can also induce production of soluble innate defense factors,
such as IgA and cytokines and microbicidal defensin proteins by Paneth cells in the small
intestine. In addition, the presence of the microbiota seems to render the macrophages more
active and to promote a general pro-inflammatory state, favoring the production of the pro-
inflammatory cytokines, such as TNF and the chemokine MCP-1. Innate immune responses
can be rapidly called into action in defense of the host and then deactivated just as rapidly
when a threat is removed.

ii) THE ROLE OF ACQUIRED IMMUNITY IN PROBIOTIC ACTION

Probiotics can enhance acquired immunity. It has been shown that microbes can have
an adjuvant effect; they enhance and the
immune response to other antigens. Hence,
probiotics may modulate the acquired immune
response to other antigens in the environment
through interactions with the innate immune
system. Macrophages and dendritic cells,
components of the innate immune system,
interact with microbes in the body and process
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and present antigens to T helper cells. T helper cells determine the immune response to an
antigen. Depending on the microbe with which they interact, macrophages and dendritic cells
are induced to produce either T helper cell stimulatory cytokines (like IL-1 and IL-12) or T
helper cell inhibitory cytokines (like IL-10). Therefore, different probiotics may be used to
modulate the action of the antigen presenting cell

REDUCING ALLERGIC DISEASES

Establishment of normal gut flora, which starts after birth and continues up 1- 2 years
of age, may be important in the development in later life of counter regulatory ability against
several specific immune responses. The normal flora of the GI tracts
enters the body through food, water, air and other environmental
sources. Raising infants in an over sanitary environment and feeding
semi sterile processed foods may interfere with the establishment of
normal micro flora in the GI tract. This may cause the immune system
of infants to develop inflammatory response to many food antigens.
Probiotic containing beneficial gut bacteria can have a suppressive
effect to such reaction by stimulating the production of anti inflammatory cytokines and
reducing allergic reaction in sensitive individuals.

PROBIOTICS AS VACCINE CARRIERS FOR INFECTIOUS AGENTS

Since most probiotics are harmless natural inhabitants of GI and urogenital tracts, it is
intriguing to use them as vaccine carriers to prevent pathogens colonization / infection in
host. In an attempt to prevent HIV infection, scientists cloned HIV-specific CD4 receptor in
Latobacillus jensenni isolated from vagina. Normally
HIV surface protein, gp120 interacts with CD4
receptor of host cells to initiate infection.
Lactobacillus expressing CD4 receptor protein in
vaginal tract will interact with HIV virus thus
preventing infection in host. A modest effectiveness of
controlling infection has been demonstrated in
laboratory experiment. Harmless probiotic strain of E.
coli was also used to express enterotoxin binding
lipopolysaccharide through cloning. This probiotic bacterium showed reduced diarrheagenic
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action when challenged with toxin from toxigenic e. coli or Vibro cholera in experimental
model. The use of probiotics as vaccine carriers against in infections diseases is an exciting
and promising filed of study; however there is no such vaccine commercially available today.

PROBIOTICS IN PREGNANCY WOMEN

Good nutrition during pregnancy improves the chances of having a healthy baby who
will be at lower risk of diseases later in life. Bacterial vaginosis, has been suggested as a
factor that increases risk of preterm labour and infant mortality and probiotics been shown to
decrease risk of bacterial vaginosis and maintain normal lactobacilli vaginal flora. In animal
studies, these strains were found to be safe during pregnancy and to enhance the health of
mothers and newborns. Another area of interest in the use of probiotics in pregnancy is to
prevent allergic reactions. Studies using L rhamnosus GG and B lactis BB12 have shown that
atopic dermatitis, a condition that causes severe skin rashes in up to 15% of babies, can be
prevented in 50% of cases if mothers ingest probiotics during pregnancy and newborns ingest
them during the first 6 months of life. Probiotics during pregnancy also have an excellent
safety record (Table 5).
Table 5: Probiotics for newborns and pregnant women:
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Other than the above mentioned, there are several reports given that the
effect of fermented milk to prevent constipation by using L. acidiophilus,
L. casei, Lactobacillus GG. And it also prevent
 LIVER CIRRHOSIS, CROHN’S DISEASES
 POUCHITIS, IRRITABALE BOWEL SYNDROME, etc,.
Some common role of probiotics in human health was described
in the Table 6.
Table 6: Clinical effects of some Probiotics
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PROBIOTIC PRODUCTS
Probiotics products for human consumption fall into three categories:
1) Infant foods, 2) Culture milk, 3) Pharmaceutical preparations
There has been a tremendous commercial interest in the incorporation of
Bifidobacteria into baby foods and also in the development of “bifidus milk” contanign
growth promoting bifidogenic factors such as lactulose and fructo-oligosaccharide. Products
that have been developed for the baby food such as Lactana-B, Femilact. (Table 7).
Many products of dairy origin contains probiotic bacteria are on
the market, these include sour cream, ice cream, butter milk, yoghurt,
powdered m ilk and frozen desserts. The probiotic culture may be added
during fermentation with normal yoghurt starters, to the final fermented
products, or to the fresh product before shipment. For example “Cultura”
made in Denmark, is made by fermenting protein – enriched whole milk
with B. Bifidum and L. acidiophilus and has a shelf life of at least 20 days
after production. A drink is obtained by fermenting partially skimmed milk. ‘Biograde’,
which is produced by more than 45 dairies in Germany, is made by fermenting milk with S.
thermophilus, B. bifidium and L. acidophilus etc., which are clinical proven health benefits.
Table 7: Some familiar commercial dairy products containing probiotic bacteria

In addition to food products containg probiotic bacteria there are various health food
adjuncts and pharmaceutical preparations contatning probiotics in the market (Table 8).
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These are generally encapsulated freeze-dried bacterial preparations, and used in the
treatment of gastrointestinal disturbances, constipation etc.
Table 8: Some commercial pharmaceutical preparations

NON DAIRY PROBIOTIC FOOD PRODUCTS

Dairy products are among the leading causes of food allergies, and there are growing
numbers of people who seek to avoid them for that reason, or because their bodies are lactose
intolerant, or because they seek to avoid all foods that come from animal sources as part of a
strict vegetarian diet. Another concern is recombinant bovine growth hormone (rBGH),
which is administered to an estimated 30 percent of conventional dairy cows to increase
production. Some scientists believe that consuming dairy products from rBGH-treated cows
may increase the risk of prostate and pre-menopausal breast cancer.
 Edensoy and White Wave's Silk soya milk, which has a smooth taste and even mixes
well in coffee. It also comes in vanilla and chocolate flavors
 Non-dairy cheese - TofuRella cheddar flavor is low in calories and fat compared to its
dairy counterpart.
 Jalapeno Jack Rice Cheese, made from rice milk, is quite tasty. It is low in fat and
calories. The very edible and tasty in sandwich and pizza.
 Poi is the starchy paste of taro corm and primarily provides carbohydrates as well as
other nutrients. Poi is made by cooking, crushing, and pounding the taro corms into a
paste by adding water and lactic acid bacteria.
Poi has some potential advantages compared to yogurt in therapeutic use in cancer
patients. Although the role of an alkaline ash diet in the treatment of cancer is debatable, poi
contributes to an alkaline diet since its calories are primarily derived from starch rather than
sugar and protein. Also, poi is easily digestible due to its very small starch granule size, and
poi is culturally acceptable to many people of Hawaiian or Polynesian ancestry.
Barry Callebaut develops probiotic chocolate with two microencapsulated specific
strains (Lactobacilli and Bifidobacteria), Probiocap™. Just 13.5g of probiotic chocolate per
day is sufficient to ensure the balance of the intestinal
microflora. Studies reveal chocolate to be a better carrier for
intestinal delivery of probiotics, as compared to dairy
products, which are normally used as carriers for probiotics.
Initial studies also confirm that chocolate is a superior carrier
for the intestinal delivery of probiotics, ensuring the survival
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of up to 4 times as many beneficial probiotic bacteria as those contained in milk-based

substrates.
In case of cereal based dairy
alternative oats milk, oats yogurt was
prepared (Eg; Yosa®, Velle®) which is rich
in rich in fiber and beta-glucanand reduce
the risk of heart disease. The oats with rich
nutrient that support the growth of the
probiotic incorporated in the fermented foods (Table 9).
Table 9: Some non-dairy probiotic foods

LEGISLATION, SAFETY CONSIDERATIONS AND CLAIMS FOR THE USE OF

PROBIOTICS IN FOODS
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Although LAB have long term history of safe use in human foods its is generally accepted
that zero risk can never be applied to the investigation of live microbes whether these are
probiotics, starters or components of natural microbiotia if fresh foods e.g. fruits and
vegetables.

Therefore even among a group of bacteria that is generally recognized as safe (GRAS) as for
example Lactobacillus spp., it is recommended that probiotic stains be characterized at a
minimum with the following tests (FAO/WHO, 2002).

 Determination of antibiotic resistance patterns

 Assessment of certain metabolic activities (e.g. deconjugation of bile salts)
 Testing foe toxin production
 Haemolytic potential
 Lack of infectivity by a probiotic strain in immunocompromised animals
 Assessment of side-effects during human studies (i.e. systemic infections, deleterious
metabolic activities, excessive immune stimulation in susceptible individuals and
gene transfer).
 Epidemiological surveillance of adverse incidents in consumers (post market).

General health claims allowed for foods containing probiotics fall under the guidelines for the
use of nutrition claims adopted by the Codex Alimentarius since 1997, FAO/AHO (2002),
has recommended that specific health claims such as enhanced function and/or disease
prevention claims on foods containing probiotics should be allowed only in cases where
sufficient scientific evidence is available. In the European Union such evidence has to
assessed by the European Food Safety Authority, prior to its use in the labeling, presentation
and advertising of foods, according to the European Commission 2003/0165 (COD) proposal
on nutrition and health claims made on foods.

QUALITY CONTROL OF PROBIOTICS IN FOOD

As of August 2007, there are no requirements for a minimal number of
microorganisms in foods in order for those foods to be classified as “probiotic foods” in the
USA. Certain food associations have made recommendations for the number of organisms
and/or the type of microorganisms in their food products. For example, the National Yogurt
Association recommends the minimal presence of 10 8 viable organisms per gram of a yogurt
product at production. To encourage compliance among manufacturers and confidence from
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consumers, the National Yogurt Association will add a seal (Live Active Seal) to products
containing this minimal count of microorganisms. The quantification of microorganisms
within a product must be certified by independent microbiologists to qualify for the Live
Active Seal. The National Yogurt Association however, does not specify whether these
organisms are “probiotic organisms” or organisms such as L. bulgaricus and Streptococcus
thermophilus that are used as “starter cultures”
RECOMMENDATIONS FOR CLINICIANS
What recommendations can we make to healthcare professionals and their patients
when choosing a probiotic that will be safe? The first and foremost step is to check the peer-
reviewed literature and consumer websites. These resources provide clinicians and patients a
way to check on the quality control of probiotic foods and supplements. Specifically, these
resources indicate which brands have been tested for purity, viability, and efficacy with
regard to immune support or specific health care problems. The most commonly used sites
are listed in Table 10.
Table 10: Websites to investigate quality control of
probiotics

Current Status of probiotics in India

In India, only sporulating lactobacilli are produced and they are prescribed with some
of the antibiotic preparations. Here probiotics are often used as veterinary and poultry feed
supplements. This requirement is also met by importing probiotics from other countries. It is
rarely used for human beings – Sporolac, Saccharomyces boulardii and yogurt (L. bulgaricus
+ L. thermophillus) are the most common ones. Sporolac is manufactured using
Sporolactobacilli. Lactobacilli solution is an example of a probiotic, usually given to
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pediatric patients in India. The latest and recent addition to the list of probiotics in India is
ViBact which is made up of genetically modified Bacillus mesentricus. This acts as an
alternate to B-complex capsules.
FUTURE PERSPECTIVES
 Genetically modified lactic acid bacteria have been proposed as a vehicle to deliver
vaccine in the gastrointestinal tract.
 Various processing advances, such as microencapsulation and bacterial coating and
addition of perbiotoic compounds used as growth factors for probiotics organism,
will help in survival of strain.
 Characterization of the possible antimicrobial properties and their metabolic
activities.