FT – 549 – Recent Trends in Fermentation
Technology
PROBIOTICS IN VARIOUS FOODS:
FERMENTED PRODUCTS
BY
SRIRAM M (FEM21004)
M. Tech in Food Engineering and Technology
Department of Food Engineering and Technology
Tezpur University, Tezpur, Assam

Definitions
The World Health Organization defines that –
 Common definition – Probiotics defined as micro-organisms that have a beneficial
effects on the host intestinal microbial balance.
 Probiotics organisms are the microorganisms thought to be beneficial to the host
organisms.
 A bacterial strains that -
 The most common type of these beneficial bacteria are Lactobacillus and
Bifidobacterium.
Probiotics as “live micro-organisms which, when administrated adequate
amounts, confer health benefits on hosts”
• Survive stomach acid and bile
• Adhere in intestinal lining
• Grows and establish temporary resistance in intestine
• Import health benefits

History of Probiotics
Year Authors and their discovery/finding
1857 Louis Pasteur discovered lactic acid bacteria (Potential benefits of Lactobacillus)
1900 Moro described the Bacillus acidophilus
1905 Russian noble prize winner and father of modern immunology, Metchnikoff Elie, a scientist at the
Pasteur Institute was the first conceptualize the term “Probiotics”
1907 Metchnikoff proposed that the acid producing bacteria in fermented milk products could prevent
“fouling” in the large intestine (Health benefits of Lactic acid bacteria)
1965 Lilley and Stillwell coined the term “ Probiotics”
1980’s Fuller establishes first definition of probiotics – A live microbial feed supplement beneficially affects
host micro-organisms by improving its intestinal microbial balance
1991 Henry Tissier isolated the Bifidobacterium
The root of the word ‘Probiotics’ comes from the Greek word Pro means ‘Promoting’ and biotic means ‘Life’.
Metchnikoff Elie
(1846-1926)
Louis Pasteur
(1822-1895)

Characteristics of Probiotics
Probiotic micro-organisms are:
 Able to survive the passage through the digestive system
 Able to attach to the intestinal epithelia and colonize
 Able to maintain good viability
 Able to utilize the nutrients and substrates in a normal diet
 Non-pathogenic and non-toxic
 Capable of exerting a beneficial effect on the host
 Stability of desired characteristics during processing,
storage and transportation
 Anti-inflammatory, anti-mutagenic & immunostimulatory
Advantages
 Producing lactic acid: lowers the pH of the intestine and
inhibits bacterial villains such as E. coli, Salmonella &
Clostridium.
 Aid absorption of minerals, especially calcium, due to
increased intestinal acidity.
 Produce a wide range of antimicrobial substance
acidophilin and bacitracin etc. helps to control pathogenic
bacteria
 Acts as barrier to prevent harmful bacteria from colonizing
the intestine
Mode of action of Probiotics
 Production of inhibitory compounds
 Competition for chemical/available energy
 Enhancement of the immune response
 Improvement in water activity
 A source of macro and micro-nutrients
Mechanism of action of Probiotics

Most common Probiotics strains
Types of Lactobacillus species
Types of Bifidobacterium species
Streptococcus species

Probiotics Food Products
Dairy Probiotic Products
 Fermented milks and yogurt (bio-yogurt)
probiotic products.
 Ice-cream and frozen probiotic products.
 Cheese probiotic products.
Non-dairy Probiotic Products
 Fruits and vegetable probiotic products.
 Cereals and soya based probiotic products.
 Meat probiotics products.

Dairy Probiotic Products
Fermented milk and yogurt (bio-yogurt) probiotic products
 Fermented milks refers to those milk which have been made by employing selected micro-organisms to develop
the characteristics flavor and/or body and texture.
 Types: Natural butter milk, cultured buttermilk, Acidophilus milk, Bulgarian buttermilk, Kumiss, Kefir, Yoghurt
and Dahi.
 Starter Propagation: Starter species most commonly used in the industry are: Str. lactis, Str. diacetilactis, Leuc.
Citrovorum and Leuc. dextranicum.
Natural Buttermilk
This is a by products of churning cream for butter making. Ripened cream which undergoes a clean, lactic
fermentation is usually preferred. In Indian condition, a country buttermilk obtained as a by-products of churning
whole milk curd for production of country butter.

Cultured Buttermilk
 This is obtained by inoculation and incubation of pasteurized skim
milk with lactic starter.
 Cream is a starting substrate, butter is normally made by churning
the cream that has been sourced by lactic acid bacteria.
 Sour cream uses Str. cremoris or Str. lactis to producing lactic
acid and Leuc. cremoris for characteristic flavor.
 Ripened cream uses Str. cremoris or S. lactis to producing lactic
acid rapidly and Leuc. citrovorum produce the necessary flavors.
Acidophilus milk
 It is obtained by whole milk or skimmed milk to be fermented with
a culture of Lactobacillus acidophilus.
 It is claimed that acidophilus milk has therapeutic and health-
promoting properties.
 It has as an overall beneficial effect on people especially those
suffering from frequent diarrhoea and intestinal gas.
 When this milk is consumed, the beneficial bacteria was activated
in the worm stomach and intestinal tract.

Bulgarian Buttermilk
 Bulgarian sour milk comes under the category of yoghurts which
contain live bacteria. It is consider as best probiotic food.
 Usually, a pure single strain of Lact. bulgaricus is propagated
for product manufacturing.
 Now-a-days, It is prepared by combination of two strains such as
Lact. bulgaricus and Str. Thermophilus.
 This finished product is characterized by a higher acidity (1.2-
1.5%)
Kumiss
 This is a lactic acid-alcohol fermented milk (made from mare’s
milk), Originated in Russia.
 The culture consists of Lact. bulgaricus or acidophilus.
 Kumiss (2.5%) has a higher alcohol content than kefir.
 The first incubation period is followed by the addition of yeast
and a second incubation period.

Kefir
 This is originated in the Caucasian Mountain.
 It is made with fermenting agent called ‘Kefir grains’ which contains Str. lactis,
Betabacterium caucasicum, kefir bacilli and lactose fermenting yeasts.
 It is a self-carbonated beverage containing 1% lactic acid and 1% alcohol.
Yogurt
 Yogurt, as a fermented milk product, is one of the most popular food carriers for the
delivery of probiotic.
 Yogurt has long been recognized as a product with many desirable effects for
consumers, and it is also important that most consumers consider yogurt to be
‘healthy’, add to that incorporation of probiotic bacteria, such as L. acidophilus and
B. bifidum, into yogurt may add extra nutritional and physiological values.
Yogurt is a fermented milk product that has been prepared traditionally by allowing
milk to ferment at 42–45°C.
 Modern yogurt production is a well-controlled process that utilizes ingredients of
milk, milk powder, sugar, fruit, flavors, colorings, emulsifiers, stabilizers, and
standard pure cultures of LAB (Str. thermophilus and L. bulgaricus) to conduct the
fermentation process. S. thermophilus and L. bulgaricus exhibit a symbiotic
relationship during fermentation process of yogurt, with the ratio between the species
changing constantly.
Types of yogurt
Natural set yogurt Stirred yogurt
Drinkable yogurt

Bio-yogurt
 Recently, the Bio-Yogurt products have been formulated
with different types of probiotic strains: mainly species of
Lactobacillus and Bifidobacterium include L. acidophilus;
L. casei; L. gasseri; L. rhamnosus; L. reuteri; B. bifidum;
B. animalis; B. infantis; and B. longum.
 Different types of Bio-yogurts are produced by far
including plain, stirred, flavored, and fruits added Bio-
Yogurts.
Dahi
 Dahi or curd is a product made by fermenting milk of the
cow or buffalo milk with lactic acid bacteria.
 The pH of curd is 4.5-4.7
Designation Culture used
Sweet dahi Str. lactis Single or in combination
Str. diacetilactis with or without Leuc.
Str. Cremoris species
Sour dahi Same as above, along with L. bulgaricus or Str.
Thermophilus or both

Ice-cream and frozen probiotic products
 Ice-cream is a frozen dairy product, consists of a mixture of
components, include, milk, flavoring, sweeteners, stabilizers, and
emulsifiers agents.
 During the last few decades, new type of the ice-cream products have
been introduced to the markets, these products were developed by
incorporating probiotic cultures into ice-cream products.
 The incorporation of probiotic cultures into ice-cream resulted in
adding value to the ice-cream product and being considered as a
functional product, in addition to being a rich food from the nutritional
point of view, containing dairy based material, vitamins and minerals in
its composition.
 As a result to the composition/structure, manufacturing procedures,
and storage conditions, ice-cream and frozen dairy desserts
demonstrated great potential for use as vehicles for probiotic cultures.
General steps involved in probiotic
ice-cream manufacturing
Mixing of Ingredients
(Milk, milk powder, sugar, emulsifiers,
stabilizers)
Pasteurization
Cooling around 37–40°C for the soured ice-
cream
The freeze-dried starter cultures (usually
yoghurt cultures)
The probiotic cultures is added
Cooling and keeping the mixture at 4◦C for
24h for the maturation
Probiotics Ice-cream is produced
Storage and transport
Probiotic micro-organisms in ice-cream
L. acidophilus; Bifidobacterium animalis; B. lactis; L. johnsonii; L.
rhamnosus; L. agilis; Bifidobacterium longum; Bifidobacterium
lactis; L. paracasei; Bifidobacterium bifidum; L. gasseri & L. reuteri

Amul Prolife Sugar Free Probiotic Wellness Frozen Dessert
Now with the changing trends, Amul has introduced first time in India – Amul pro-life sugar free probiotic
wellness frozen dessert: Low fat low calorie
 It has been made by reduced fat content and sugar replacement with the combination of low calorie sweetener
(Fructo-oligosaccharides and sucralose) denotes that the probiotic compounds has the most beneficial effects on
probiotic stability.
 These sweetener has very low glycemic index because they sparingly gets digested by digestive enzymes in
stomach and small intestine.
 Low Fat: Amul pro-life sugar free delight is containing 50% less fat than the normal ice-cream.
 Low Calorie: Sugar replaced with low calorie sweetener.
 Available in five different flavor – Vanilla, chocolate, strawberry, fresh litchi and shahi anjir.

Amul Prolife Probiotic Wellness Ice-cream
Again, Amul has launched, India’s First Probiotic Ice-cream aimed at the
total health conscious of consumers.
 Available in five different flavor – Vanilla, chocolate, strawberry, fresh litchi
and shahi anjir.
Frozen Yogurt
 Frozen yogurt is generally referred to as the frozen ice milk processing a
typical yogurt flavour.
 In United states, frozen yogurt is a blend of 90% ice-cream, 10% plain
yogurt with pH is around 6.
 Frozen yogurt in Netherlands must contain at least 70% yogurt and the pH is
less than 5.0. On the other hand, Australia requires in production of frozen
yogurt with greater than 45% yogurt and the pH should be less than 4.5.
 In recent years, frozen yogurt has become the base for development of other
innovative products such as frozen flavoured yogurt dessert, frozen yogurt
novelties on a stick or sandwiches.
 Probiotic organisms such as Bifidobacterium spp and L. acidophilus have
been successfully added into frozen yogurt.

Cheese Probiotic Products
 Cheese means the product obtained by draining after the
coagulation of milk with a harmless milk coagulating agents,
under the influences of harmless bacterial cultures.
 During production, the milk is usually acidified and adding the
enzymes rennet causes coagulation.
 Cheese is a concentrated form of two milk components – Milk
protein (casein) and milk fat.
Types of cheese and micro-organisms involve
1. Soft cheese – Str. cremoris; Penicillum camemberti
2. Semi soft cheese – Lactococcus lactis; Brevibacterium linens
3. Hard cheese – L. casei; L. plantarum
4. Semi hard cheese - L. casei; Str. cremoris
5. Fresh cheese– Str. sp
6. Blue cheese– Penicillum roqueforti; Lactococcus lactis
7. Processed cheese (fungi or fungal spores used during ripening

Classification of cheese

Example of Probiotic cheese
Probiotic cheddar cheese
 Cheddar cheese has a great potential as a probiotic ‘functional food’.
 Currently, these are the probiotic strains including L. acidophilus; L.
casei; B. longum; L. paracasei and B. lactis were examined as
potential candidate for incorporation in Cheddar cheeses.
 These strains have been selected based on their acid and bile
tolerance, adhesion to intestinal cell line, anticarcinogenic properties,
oxygen sensitivity and ability to modify gut microflora of human
subjects.
 In general, a probiotic cheese should have the same performance as a
conventional cheese: the incorporation of probiotic bacteria should
not imply a loss of quality of the product.
Manufacture of Probiotic Cheddar cheese
(Flow diagram of manufacturing)

Non-dairy Probiotic Products
Fruits and Vegetables Probiotic Products
 Fruits and vegetables are considered healthy foods, as they contain several
beneficial nutrients, such as minerals, vitamins, dietary fibers, and antioxidants.
Unlike dairy products, fruits and vegetables lack allergens, lactose, and cholesterol,
which adversely affect certain segments of the population.
 Moreover, recent technologies advances have made alterations to some structural
characteristics of fruits and vegetables matrices by modifying food components in
a controlled way such as pH modification, and fortification of culture media, that
might make fruits and vegetables ideal substrates for probiotic bacteria delivery to
human
 Several type of probiotic fruits and vegetables products have been developed such
as fruits and vegetables juices, dried fruits, fermented vegetables, and vegetarian
deserts.
 Wide range of probiotic strains, mainly species of Lactobacillus and
Bifidobacterium such as L. acidophilus; L. casei; L. paracasei; L. rhamnosus
GG; L. plantarum; L. fermentum and B. bifidum have been widely used in the
development of many fruit and vegetable products, specially juice products,
include orange, pineapple, cranberry, cashew apple, tomato, cabbage, beet and
carrot juices.
Gundruk
Sinki
Khalpi
Soidon

Examples of traditional fermented fruits and vegetables, which are used in various parts of Asian subcontinent

Cntd.….

Cntd.….
Kimchi Sauerkraut Pao cai
Tempoyak Sayur Asin
Nozawana zuke

Cereal probiotic products
 Cereals have complex nutrient composition and are being consumed
on a daily basis all over the world as one of the staple foods.
 Benefit of consuming fermented cereal based foods is the availability
of dietary fiber and presence of non-digestible carbohydrates like
oligosaccharides can act as a probiotic which can stimulate the
growth of probiotic LAB.
 Cereal grains like maize, sorghum, millet, oats, barley, wheat and
rye are being used for this purpose.
 Fermentation of cereals increase the bioavailability of minerals such
as phosphorous, iron and zinc due to the action of microbial enzymes
such as phytases, or due to the organic acids produced during
fermentation of cereals.
 Commercial cereal based probiotic products includes cereal-based
puddings; Rice-based yogurt; Oat-based drink; barley, and malt
based; Yosa (oat-bran pudding); Mahewu (fermented maize
beverage) Maize-based beverage
 Examples of the traditional non-dairy cereal- based fermented
beverages include Boza, Tarhana, Kishk, Chicha, Kisra,
Kenkey…etc.
Cereal and soya based probiotic products

Soya based probiotic products
 Soya, the most important legume in the traditional Asian diet, is
rich in high-quality protein. The products of soybean play an
important role in the prevention of chronic diseases such as
menopausal disorder, cancer, atherosclerosis, and osteoporosis.
Soya probiotics products include, soymilk, soy-based yogurt,
vegetarian frozen desert, fermented soy tempeh, and soy cheese,
are a good substrate for the growth of probiotic bacteria.
 Soy yogurts were prepared with a yogurt starter in conjunction
with either the probiotic bacteria L. johnsonii, L. rhamnosus
GG or human derived Bifidobacterium.
 Probiotic frozen vegetarian soy desserts were developed with
the incorporation of L. acidophilus, L. rhamnosus, L. paracasei
ssp. paracasei, Saccharomyces boulardii and B. lactis.
 The soy cheese was made from soymilk fermented with soy
cheese bacterial starter cultures and L. rhamnosus.

Meat probiotic products
 Meat is a highly nutritious food with a high degree of nutrients bioavailability and
consumers have a high degree of preference for its taste, flavor, and texture. Meat had
shown an excellent vehicle for probiotics as a result to meat composition and structure.
Meat was found to have a protection effect on LAB against the lethal action of bile.
 The most common processed probiotic meat products is the dry fermented sausages
without heating.
 A raw cured sausage is a mixture of different meat grades preserved by curing, stuffed
into casings and microbiologically fermented with salt, nitrate or nitrite, sugar and
spices (mostly garlic and black pepper) added.
 Salami-type raw cured sausages are a wide group of products which differ in raw
material composition, ripening time, external appearance and flavor. The original
recipe for the salami-type raw cured sausage, initially made from donkey meat, was
developed in Italy
Lactic acid bacteria: L. plantarum; L. sake; L. pentosus; L. casei; L. curvatus; L.
alimentarius; Pediococcus acidilacti; Pediococcus pentosaceus.
Gram-positive cocci: Staphylococcus carnosus; Staphylococcus xylosus;
Micrococcus varians.
Yeasts: Debaryomyces hansenii; Candida famata.
Moulds: Penicillium nalgiovense; Penicillium camamberti; Penicillium chrysogenum
Types of fermented meat
products
Hams
Loins
Sausage

Examples of potential utilization of probiotic bacteria
in meat products

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