Milk Fermented Products 2013

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Critical Reviews in Food Science and Nutrition , 53:482–496 (2013)

Copyright C Taylor and Francis Group, LLC

ISSN: 1040-8398 / 1549-7852 online

DOI: 10.1080/10408398.2010.547398

F e r m e n t e d M i l k s a n d M i l k P r o d u c t s

a s F u n c t i o n a l F o o d s — A R e v i e w

V. K. SHIBY1 and H. N. MISHRA2

1Freeze Drying and Animal Products Technology Discipline, Defence Food Research Laboratory, Siddharthanagar, Mysore,

India 5700112Department of Agricultural and Food Engineering, Post Harvest Technology Centre, Indian institute of Technology,

Kharagpur, India

Fermented foods and beverages possess various nutritional and therapeutic properties. Lactic acid bacteria (LAB) play

a major role in determining the positive health effects of fermented milks and related products. The L. acidophilus and

Bifidobacteria spp are known for their use in probiotic dairy foods. Cultured products sold with any claim of health benefitsshould meet thecriteria of suggested minimum number of more than 106 cfu/g at the time of consumption. Yoghurt is redefined

as a probiotic carrier food. Several food powders like yoghurt powder and curd (dahi) powder are manufactured taking into

consideration the number of organisms surviving in the product after drying. Such foods, beverages and powders are highly

acceptable to consumers because of their flavor and aroma and high nutritive value. Antitumor activity is associated with the

cell wall of starter bacteria and so the activity remains even after drying. Other health benefits of fermented milks include

prevention of gastrointestinal infections, reduction of serum cholesterol levels and antimutagenic activity. The fermented

products are recommended for consumption by lactose intolerant individuals and patients suffering from atherosclerosis. The

formulation of fermented dietetic preparations and special products is an expanding research area. The health benefits, the

technology of production of fermented milks and the kinetics of lactic acid fermentation in dairy products are reviewed here.

Keywords Fermented milks, L. acidophilus, bifidobacteria, probiotics, lactic acid bacteria

INTRODUCTION

Healthiness is one of the most frequently mentioned reasons

behind food choices in EU countries (Lappalainen et al., 1998).

Individuals are considered responsible for their own state of

health. Diseases are seen more and more as consequences of

consumer’s own behavior rather than the result of environment

(Ogden, 1998). Diet may strongly influence an individual’s risk

of obesity; cardiovascular diseases, cancer, and other life style

related diseases. Traditionally, the healthfulness has been as-

sociated with nutritional factors such as fat, fiber, salt, and vi-

tamin contents of food. In addition to traditional healthinessfood may contain single components that may have a positive

impact on our well-being. Products that are claimed to have

special beneficial physiological effects in the body are usually

called functional foods (Bellisle et al., 1998; Roberfroid, 1999;

Hardy, 2000; Kwak and Jukes, 2001). A particular food may be

made more functional by increasing or adding a potential health

Address correspondence to Prof. H. N. Mishra, Department of Agriculturaland Food Engineering, Post Harvest Technology Centre, Indian Institute of Technology, Kharagpur-721302, India. E-mail: [email protected]

promoting entity. Alternatively, concentration of adverse com-

ponents may be reduced or there may be a partial interchange

between toxic and beneficial ingredients. Functional food may

be considered as a therapeutic aid without prescription. The

term nutraceutical was coined by Stephen De Felice, founder of

the Foundation for Innovation in Medicine, in 1989. He defined

a nutraceutical as a food, dietary supplement or medical food

that has a medical or health benefit including the prevention or

treatment of disease. Today the term nutraceutical is often used

interchangeably with functional food. An official definition for

the term functional foods is lacking in USA and Europe. Ac-

cording to EU concerted action “a food can be considered asfunctional if it has been satisfactorily demonstrated to affect

beneficially one or more target functions in the body beyond

adequate nutritional effects in a way that is relevant to either

an improved state of health and well-being and/or a reduction

of risk of disease.” These functional foods may also be used as

conventional foods (Diplock et al., 1999).

Thereare several methods of manufacturing functional foods,

based either on the method used to produce them or on their

purpose. Functional foods may be processed by modification

or they may be fortified with different substances and the

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FERMENTED MILKS AND MILK PRODUCTS AS FUNCTIONAL FOODS—A REVIEW 48

functionality of a product can be targeted to a special disease or

just to improve overall well being (Roberfroid, 1999). Informa-

tion relating health effects and the ways of communicating such

information are the key factors because the health effect cannot

be perceived from the product itself (Urala et al., 2003). Since

1989, the nutraceutical or functional food industry has evolved

into a market worth $20.2 billion in 2002. Functional beveragesreached$ 10.35 billion sales in 2002, up 10.7% andare projected

to reach $15.9 billion by 2010. A variety of functional dairy

foods are conquering the market worldwide. Examples include

probiotic, prebiotic, and symbiotic products, low cholesterol

fresh milk omega-3-milk, low lactose, and lactose free products

and milk products that can control or manage hypertension and

immune functions. The market for functional foods, nutraceu-

ticals, wellness foods, and beverages is fast growing and it is

estimated that the functional food industry could almost dou-

ble by 2007. An increasing sector of the public is quickly aging

and purchasingfunctional food products. Today’s consumers are

more concerned with weight loss and management, heart health,

eye health, cancer, increasing energy and stamina, eye health,

and improved memory. Several categories of functional foods

and beverages have been developed to meet these challenges.

The scope of the present article is to provide an overview

of the kinetics of lactic acid fermentation in dairy products,

the technology and nutritional significance of fermented milks

popular in different countries, and the therapeutic properties of

probiotic organisms involved in their manufacture.

LACTIC ACID FERMENTATION

Fermentation Kinetics

Mathematical analysis of the biokinetics of functional starter

cultures can yield precious information about the relationship

between the food environment and bacterial functionality and

may contribute to optimal strain selection and process design.

This may result in better process control, enhanced food safety

and quality, and reduction of economic losses. Kinetic data are

needed to understand the fermentation process and to develop

continuous fermentation systems. Lactic acid bacteria (LAB)

have long been used in food industry as starter cultures for

the manufacture of fermented meat and dairy products. The

key metabolite produced during such fermentation reactions is

lactic acid, which is a commercially valuable product with ap-

plications in food manufacturing and pharmaceutical industries.

Both chemical and biotechnological methods are available for

lactic acid production, but the lactic acid from fermentative pro-

cesses has increased its market share owing to the preference

of consumers to products of natural origin (Chahal, 1989). The

commercial significance of dairy fermentation industry which

incorporates production of cheeses, fromage frais, sour cream,

etc. is well recognized and ranks second only to the alcoholic

beverages (Daly et al., 1998). The processes for wood utiliza-

tion can be directed toward lactic acid production (Moldes et al.,

1999). The kinetics of milk fermentation by starter cultures

has been studied by several research workers. Response Sur-

face Methodology has been successfully used in modeling o

acidification rate and growth of starter bacteria as a functio

of parameters such as fermentation temperature, fat and soli

content, inoculum size, and cocci/rods ratio on the fermenta

tion process of yoghurt (Torriani et al., 1996). The pH mea

surement method described by Spinnler and Corrieu (1989)

widely used in the determination of acidification kinetics duringel formation. Several research workers have tried to optimiz

the acidification process for manufacture of fermented prod

ucts and developed models to predict the acidification kinetic

Mathematical models have been used to predict the influenc

of fermentation operating temperatures on cell growth rate, ce

concentration, substrate utilization rate and lactic acid produc

tion rate. As for models, a number of them, purely or partiall

empirical, have been described in literature. Product inhibitio

is well-established feature of lactic fermentations.

Boonmee et al. (2003) developed a model for batch an

continuous fermentation of Lactococcus lactis strain NZ133

The growth kinetics of L. lactis is predominantly influenced b

lactose limitation and lactate product inhibition with the growt

of this particular strain showing a relatively high sensitivity t

lactate inhibition. The Leudeking-Piret equations for growt

and lactate production were successfully incorporated into th

model. The Leudeking–Piret model is given as

dx

dt = µx (1

dp

dt = α

dx

dt + βx (2

When this is modified to include the effects of lactose limtation and inhibition, as well as lactate inhibition the model

outlined as in Equatons (3)–(5).

The rate of biomass production

dx

dt =

s

Ksx + s

1 −

P − P ix

P mx − P ix

×

Kix

Kix + s

x (3

The rate of lactate production

dp

dt = α

dx

dt + qp,max

s

Ksp + s

×

1 −

p − P ip

P mp− P ip

Kip

Kip + s

x (4

The rate of lactose utilization

ds

dt = qs,max

s

Kss + s

1 −

p − P is

P ms − P is

×

Kis

Kis + s

x (5

where,

K i is lactate inhibition constant (gl−1),

K s is lactose limitation constant (gl−1),


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484 V. K. SHIBY AND H. N. MISHRA

P is lactate concentration (gl−1),

Pi is threshold lactate concentration (gl−1),

Pm is maximum lactate concentration (gl−1),

q p,max is maximum specific lactate production rate (gg−1h−1),

qs,max is maximum specific lactose utilization (gg−1h−1),

s is lactose concentration (gl−1),

x is biomass concentration (gl−1),

a is growth associated constant in Leudeking–Piret model

(gg−1),

b is nongrowth associated constant in Leudeking–Piret model

(gg−1h−1),

µ is specific growth rate in Leudeking-Piret model (h−1), and

µmax is maximum specific rate (h−1)

Common Organisms and Pathways

The group of LAB plays a central role in fermentation pro-

cesses, and has a long and safe history of application and con-sumption in production of fermented foods and beverages. (Ray,

1992; Wood and Holzaphel, 1995; Wood, 1997; Caplice and

Fitzgerald, 1999) During fermentation of lactose to lactic acid

the acidity of the milk rises and the growth conditions for mi-

croorganisms other than LAB become increasingly unfavorable

(Driessen and Puhan, 1988). Reid et al. (2002) have studied the

LAB in fermented foods in South East Asia. They discussed

the distribution and application of LAB in various products

such as fermented milk, meat, fish, vegetable, or plant prod-

ucts. Significant advances have been made in elucidating the

genetic, biochemical land physiological basis of many of the

key technological traits of these bacteria (Daly et al., 1998).

LAB are constituted of heterogenous group of Gram-positivebacteria with a strictly fermentative metabolism (Temmermman

et al., 2004). One of the most important functional properties

common to all LAB is their ability to convert lactose and a va-

riety of other carbohydrates into the primary end product, lactic

acid, by fermentation. The ability of LAB to transform food

into new products and to exert antagonistic action toward harm-

ful microorganisms make them suitable for both domestic and

industrial productions Servin (2004). The LAB involved in the

production of fermented milks are distributed in several phe-

notypic and genotypic groups. These groups are characterized

by different nutritional requirements, metabolic, cultivational,

and technological properties. Several authors have reported that

LAB present in fermented milks may belong to species of Lac-

tobacillus, Streptococcus, Pediococcus, Leuconostoc, and Lac-

tococcus genera (Aucloir and Mocquot, 1974 ; Cogan, 1985;

Gilliland, 1985; Sandine, 1985; Schleifer et al., 1985). The role

of interaction between yeasts and LAB in African fermented

milks has been reviewed by Narvhus and Gadaja, 2003.

There are two main fermentation pathways in LAB; the ho-

molactate pathway in which lactic acid is the major end product

and heterolactate pathway in which other compounds such as

acetic acid, carbon dioxide, ethanol etc. are produced in addi-

tion to lactic acid (Huggenholtz, 1993;Cocaign-Bous-quet et al.,

1996; de Vos, 1996). The homofermentation and heterofermen-

tation pathways are shown in Fig. 1. Both rods and cocci follow

different pathways of carbohydrate fermentation and achieve

end products, which are at least 90 or 50%, lactate in the case

of homo and hetero fermentative species respectively. Lactose

is transported across the cell membrane either by lactose per-

meate system or by phosphoenol pyruvate dependent phosphotransferase system (Lac PEP-PTS ). Depending on the transport

mechanism involved, unphosphorylated lactose (Lac permease)

is hydrolyzed into glucose and galactose by β-galactosidase

or phosphorylated lactose is hydrolysed by phosphorylated β -

galactosidase into glucose and galactose-6-phosphate. All of

the dairy lactococci and some mesophilic lactobacilli oper-

ate the Lac PEP-PTS system (Fig. 2) while leuconostoc and

some industrially significant thermophilic organisms; S. ther-

mophilus and Lb delbrueckii subsp. bulgaricus possess a perme-

ase system (Fig. 3) Subsequent metabolic pathways include the

glycolytic Embden-Meyerhof-Parnas(EMP), the phosphoketo-

lase pathways for glucose, the tagatose-6-phosphate pathway

for galactose-6-phosphate and the Leloir pathway for galac-

tose (when it is not transported back into the environment).

LAB containing aldolase, that is, Streptococci, Pediococci, and

obligately homofermenative and facultative heterofermentative

Lactobacilli have the homolactic fermentation with lactate as the

exclusive end product. LAB containing phosphoketolase can be

divided into twogroups. Thefirst, that is, Leuconostocs and obli-

gately hetero fermentative lactobacilli, follows “6P—gluconate

pathway” with production of equimolar amounts of CO2, lac-

tate and acetate. The others follow “bifidus pathway” with the

formation of acetate and lactate in a molar ratio of 3:2.

Among the LAB, lactose metabolism in lactococcus is prob-

ably the best understood in terms of its biochemical and ge-netic make-up and also in terms of its regulation. The complete

metabolic pathway is known and several important genes have

been characterized (de Vos and Vaughan, 1994). The genes en-

coding the PEP-PTS system and the tagatose pathway are all

plasmid located in a single operon under the control of the

repressor gene lacR. Another operon has also been identified

in Lactococcus encoding a number of glycolytic enzymes The

fermentative action of specific LAB strains may lead to the re-

moval of toxic or antinutritive factors such as galactose and

lactose from fermented milks to prevent lactose intolerance and

accumulation of galactose (Wouters et al., 2002).

Fermentation Process and Health

Fermented milks constitute important part of our food. In an-

cient days, human beings realized that fermentation occurred,

without, however, knowing their cause. Originally milk fer-

mented spontaneously, and the reuse of fermentation vessels

and tools contributed to a certain repeatability and stability in

the fermentation process. This led to the use of specific microor-

ganisms for the manufacture of more or less refined products.

Different countries or even different parts of the same country

developed their own fermented milks. The best-known product


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Glucose

Fru-1,6 P Glc-6PFru-6P

Acetyl-P+ Erythrose-4P6P-Gluconate

Heptose-P Xylulose-5P +CO2

+Pentose-P

Triose-3P

Phosphoketolase

aldolase

Triose-3P + Acetyl- P

PyruvatePyruvatePyruvate

Acetyl-P+ Triose-3P

ADP

ATP

ADP

Lactate LactateAcetate (Ethanol )

Lactate Acetate

GlycolysisBifidus pathway 6P-Gluconate pathway

Phosphoketolase pathways

HOMOFERMENTATION HETEROFERMENTATION

Fru-6P

Figure 1 Main pathways of hexose fermentation in lactic acid bacteria (Kandler, 1983).

is the thermophilic fermented milk, yoghurt, which has enjoyed

increased popularity in the last three decades. The role of living

microorganisms in fermented dairy products has gained consid-

erablyincreased interest forboth theconsumerand theproducer.

Nature has provided a certain degree of beneficial association

for humans through the activity of lactic LAB. These bacte-

ria are widely used in foods and agricultural fermentations. In

addition to their preservation, nutritional and therapeutic im-

portance, these are known for their longevity of human life

(Ram and Bhavadasan, 2002) LAB are natural inhabitants o

gastrointestinal tract. These microorganisms have a number o

traits which make them particularly attractive to be used a

“probiotic”. During fermentation of lactose to lactic acid th

acidity of milk rises and the growth conditions of microorgan

isms other than LAB become increasingly unfavorable. Apa

from the main metabolite lactic acid, fermentation microorgan

isms also produce bacteriostatic compounds. It has been sug

gested that the ingestion of LAB may counteract the effect o


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Figure 2 Scheme of lactose and galactose utilizati on by lactobacilli and streptococci (Kandler, 1983).

Escherichia coli outgrowth by a number of possible mechanisms

such as anti- E. coli metabolites, detoxification of enterotoxins,

prevention of toxic amine synthesis, andadhesionto thegut, thus

preventing colonization by pathogenic bacteria. These aspects

are comprehensively reviewed by Gurr (1983). Homofermenta-

tive LAB convert the available energy source (Lactose) almost

completely to lactic acid via pyruvate to produce energy and to

equilibrate the redox balance. The fermentation action of spe-

cific LAB strains may lead to removal of toxic or antinutritive

factors, such as lactose and galactose, from fermented milks to

prevent lactose intolerance and accumulation of galactose.

Culture Containing Milk Products in Prevention of Gastro

Intestinal Infections

The beneficial effects of LAB and cultured milks have been

attributed to their ability to suppress the growth of pathogens

either directly or through the production of antibacterial sub-

stances such as lactic acid, peroxide, and bacteriocins (Kodama,

1952; Kansal, 2001). LAB are credited with imparting a number

of nutritional and therapeutic attributes to fermented milk prod-

ucts such as improved digestion and bio availability of milk

constituents, inhibition to harmful bacteria of gastrointestinal

tract, suppression of cancer, alleviation of lactose intolerance,

and hypocholesterolaemic effect (Mathur et al., 2000).

Supplementation of infant formulas with Bifidobacterium

lactis and S. thermophilus has been reported to be protective

against nosocomical diahhroea in infants (Saavedra et al., 1994).

Pediatric beverage containing B. animalis, L. acidophilus, and

L. reuteri has been reported to be useful in prevention of ro-

tavirus diahhroea. Treatment with Lactobacillus GG promotes

systemic local immune response to rotavirus, which may be of

importance for protective immunity against infections (Kalia

et al., 1992). Probiotics reported to reduce antibiotic induced

diahhroea include B. longum, B. lactis, Lactobacillus GG,

L.acidophiluu La5, and Streptococcusfaecum and yeast Saccha-

romyces boulardii (Borgia et al., 1982; Colombel et al., 1987;

Surawicz et al., 1989; Nord et al., 1997). Several strains of lacto-

bacillus have been reported to inhibit Salmonella typhimurium

(Hitchins et al., 1985). Helicobacter pylori is recognized as

the main cause of anal gastritis. Inhibition of H. pylori NCTC

11637 by L. casei subsp. rhamnosus and several strains of

L. acidophilus and production of antihelicobacter factors by

these LAB have been reported (Kawamura et al., 1981; Lam-

bert and Hull, 1996).


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Figure 3 Lactose metabolism by Leuconostocs (Cogan, 1985).

TECHNOLOGY OF FERMENTED MILKS

AND MILK PRODUCTS

The nutritional importance and technology of traditional fer-mented dairy products popular in different parts of the world has

been summarized in Table 1. Besides the traditional products

which are popular in various countries, the development of pro-

biotic products, dairy beverages, dietetic preparations, and dry

cultured products are on progress due to the increasing demand

for such value added products and convenience based ingredi-

ents. The process technology for different categories of such

products is described. The major categories include cultured

milks with mesophilic organisms and thermophilic milks. The

organisms involved in their manufacture, metabolites including

flavor compounds produced and their proposed mechanism in

probiotic dairy products are given in Table 2 Dry cultured milk

products, whey based cultured beverages and several dieteticpreparations also find place in the booming health food market.

Cultured Milk

Cultured milk is a collective name applied to a wide range

of mesophilic fermented milks with a number of characteris-

tics in common. Mesophilic organisms are those with growth

optima between 20C and 30◦C. The starters used for fermenta-

tion consist of homofermentative mesophilic LAB and of aroma

producers (Driessen and Puhan, 1988). Mesophilic lactic cu

tures generally grow in the temperature range of 10–40◦C an

contain group N streptococci and/or leuconostocs (Petterson

1988). The final taste of cultured milk is the result of a mixtur

of compounds present in a certain ratio. For example, the d

acetyl flavor is associated with butter and buttermilk. Culture

milk should be mildly acid and slightly pricking. To meet thesrequirements, the content of lactic acid and carbohydrate has t

be controlled during the manufacture of the product. By coo

ing the milk at the proper time, acidification can be limited

whereas the excess of the carbon dioxide in the product at th

end of fermentation can be removed by stirring or de-aeratio

by vacuum.

Milk for production of cultured milk has to be heat treate

in order to inactivate the native substances which are inhibitor

to the LAB and to improve the structure of the final product b

denaturation of whey proteins. To obtain a coagulam that ca

be stirred easily to a smooth and viscous product, 805 or mor

of the whey proteins should be denatured (Snoeren et al., 1981

Richter, 1997) this can be reached by a heat treatment of 90◦

for 3 min or 85◦C for 30 min (Harland et al., 1955; Hillier an

Lyster, 1979).

Lowering of pasteurization temperature of milk results in de

creasing firmness and retarded acidification. Ultra High Tempe

ature (UHT) sterilized milk and other high temperature treate

milks also result in lower viscosity, age-thickening and a cooke

flavor in cultured milks. Homogenization of milk at 55◦C an

20 MPa is sufficient to get good distribution of fat. Decreas

ing milk solids-not-fat content results in taste difference. Th

product may turn “flat” and “watery” and the defect known a

“astringent” may occur. Increasing MSNF of milk leads to

“full” taste, higher viscosity, and a stable cultured milk withouwheying off during cold storage.

Dry Cultured Milk Products

Extensive research has been carried out on various dry cu

tured products including yoghurt, acidophilus milk, kumis

and kefir, usually combined with additives that will enhanc

the nutritive quality of the final product. Starters for dry cu

tured beverages are streptococcus thermophilus, Lactobacillu

bulgaricus, Lactobacillus acidophilus (LA), “Kefir grains”, an

Bifidobacterium adolescens among others (Caric, 1994). Ver

little scientific work has been published in this field and th

developed processes are covered by patents (Pallansch, 1970

Hall and Hedrick, 1975; Shah et al., 1986; Usacheva et al

1990). In general, the processing procedure for these product

differs somewhat from that for conventional cultured produc

(Krsev, 1989). Milk is heated to 90◦C–95◦C for 20 min prio

to culturing. A total solids content of 30% has been found t

be most favorable. Concentration is accomplished either b

vacuum evaporation or ultra filtration. The advantage of u

ing concentrated milk for culturing is volume reduction, an

certain stimulation is provided to microorganisms in this way


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Table 2 Probiotic microorganism, metabolites and proposed mechanism in dairy products

Product Microorganisms involved Metabolitesa,/flavorb compounds/probiotic c property

Acidophilus milk L. acidophilus Maintenance of normal intestinal florac, acidophilina

Acidophilus paste L. acidophilus

Acidophilus buttermilk L. acidophilus -do-

Acidophilus yoghurt L. acidophilus, S.thermophilus, L.

delbrueckii.subsp.bulgaricuss.

Acetaldehydeb

Biogurt L. acidophilus, S. thermophilus

Acidophilus—yeast milk L. acidophilus, lactose fermenting yeasts

Probiotic dahi L. acidophilus, L. lactis, L lactis subsp diacetylactis Diacetylb enhanced digestibility, prevents constipation

Bifidus milk Bifidobacterium bifidum, B. longum Removal of toxic aminesc, strengthening of host immune systems

Bifighurt B. bifidum, S. thermpophilus Reduction of serum cholesterol

Biogarde B. bifidum, L. acidophilus, S. thermophilus

Biokys B. bifidum, L. acidophilus, P. acidilactici

Special Yoghurt B. bifidum, L. acidophilus, S. thermophilus, L.

delbrueckii subsp. bulgaricus

Cultura B. bifidum, L. acidophilus

Cultura drink B. bifidum, L. acidophilus

Progurt S.lactis subsp. diacetylactis, S. lactis subsp.cremoris,

l. acidophilus, and/or B. bifidum

Diacetyla Allieviates lactose maldigestionc

a

Metabolites; b

flavor compounds; c

probiotic property.

However, the desired amount of inoculam varies considerably

from 1–6% according to various authors. Even starter quantities

as high as 10–15% have been reported. Various levels of lactic

acid in concentrated milk prior to spray drying were developed.

Most often acid ranges from 0.6–1.1%. Sometimes treatments

intended to minimize the heat-induced changes and to increase

starter survival during drying are carried out as well. Spray dry-

ing temperatures are lower than those commonly used for milk

drying and range from 145–175◦C (inlet air temperatures). Data

have been reported for drying cultured beverages by freeze-

drying on a laboratory scale. The resulting product is of good

solubility and contains viable microflora after recombination.Kumar and Mishra (2003) developed mango soy fortified yo-

ghurt powder. The nutritional and therapeutic value of yoghurt

was improved by supplementing buffalo milk with soymilk, thus

incorporating in the product phytochemicals. By incorporating

soymilk phenyl alanine, iron, thiamin, and niacin contents of

the product increased.

Whey Based Cultured Dairy Products

Current worldwide popularity of fruit flavored drinkable yo-

ghurt offers an excellent opportunity for incorporation of whey

into the development of cultured fruit flavored products (Singh,

2001). Proper amount of stabilizer coupled with heat processing

of the ingredients and homogenization are important aspects in

the manufacture of fermentable cultured dairy drink (Dorde-

vic et al., 1982). For making drinks from sweet or acid whey,

the standardized method involves adding sugar to filtered whey,

pasteurization at 80◦C for 5 min, homogenization at 70◦C, in-

cubation at 43◦C with 2% Lactobacillus helveticus starter for

15–17 h to the pH 3.8–3.95, repasteurization at 85◦C with 5 min

holding, addition of 4–7% fruit juice concentrate, rehomogeni-

sation, cooling to 50◦C and packing into retail containers. The

drink contained 11.65–13.1% total solids, 11.27–11.65% tota

sugar, 0.66–0.78% protein, 0.2–0.3% fat, and 0.42–0.46% min

erals. Mango-molke mix, a popular whey beverage markete

in Europe contains bifidobacteria culture along with whey an

mango juice. Probiotic attributes of LAB may be incorporate

to develop fermented whey beverages. Such beverages may b

beneficial for the people suffering from gastro-intestinal trac

disorders and it also provides nutritional and therapeutic so

drinks (Singh, 2001). Kumar and Tiwari (2003) showed tha

good quality whey based fermented milk beverage can be mad

from a mixture of cheese whey and milk. The fermented beve

age made by admixing 70% of cheese-whey, 0.3% CMC witthe milk resulting in acceptable whey based fermented mil

beverage.

Thermophilic Fermented Milks

The term thermophilic should be reserved for microorgan

isms whose optimum growth temperature lies between 55 an

75◦C. In dairy industry, it tends to be employed to describ

cultures that are most active between 35 and 40◦C. Accordin

to Bianchi-Salvadori (1981) thermophilic yoghurt starters d

not form part of the natural intestinal flora of man nor do the

implant themselves in the intestines. It has, however, been sug

gested that L. delbrueckii subsp. bulgaricus and S. thermophilu

can establish themselves in the intestine and gradually domi

nate the natural microflora. In vitro, studies have indicated tha

L. delbrueckii subsp. bulgaricus is inhibited by bile salts (Lem

bke, 1963). However, these bacteria are also able to resist a hig

degree of acidity and so it is quite feasible that a proportion o

bacteria ingested will reach the intestine in a viable state (Aco

and Labuza, 1972). Mabbit (1977) supports the hypothesis tha

certain strains of L.delbrueckii subsp. bulgaricus could surviv

and compete in the human intestine. This possibility is of grea


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importance since it would ensure the absence of putrefactive

microorganisms and hence protect the intestinal health of the

consumer (Dellaglio, 1984).

Fermented Dairy Beverages

The demand for fermented milk beverages is on the rise due

to their positive health effects and nutritive value. The general

requirements of fermented milk beverages are that they should

be light, low calorie, for example, drinking yoghurt has 30%

less energy than conventional yoghurt, refreshing (not causing

thirst), tasteful of agreeable mouth feel, and smooth in tex-

ture. Milk like soft drinks mainly prepared with yoghurt and

mesophilic starter cultures and nonsoft drinks, for example, Ke-

fir and Kumiss with alcohol production fall in the category of

fermented milk beverages. Acidified milk drinks (AMDs) need

a stabilizer, for example, high methoxy pectin if whey forma-

tion is to be avoided. A stabilizer can act in different ways.

An increased steric repulsion between the casein micelles maybe introduced, thus preventing flocculation. Alternatively, a ki-

netic barrier may be imposed against extensive aggregation.

Casein micelles in milk (pH 6.7) are generally assumed to stay

in the suspended state as the result of steric repulsive interac-

tions between the micelles. On acidification of milk to a pH

value around 4, as is done during the preparation of yoghurt,

the native mechanism of stabilization of casein micelles fails.

This failure is believed to be related to the collapse of the ex-

tended conformation of κ-casein chains (Holt, 1982 and De

Kruif, 1998). At pH 6.7 these chains protrude from the surface

of the casein micelle in order to maximise their entropy. This

same tendency to maximize entropy of κ -casein chains causes

the micelles to have a repulsive mutual interaction because over-lap of κ -casein chains of neighboring micelles would result in a

loss of entropy of the chains. This phenomenon is called steric

stabilization. In order to stabilize an AMD of 8.5% milk solids

(not fat), high methoxy pectin has to be added at concentrations

around 0.3% w/w (Trompa et al., 2004). It was found that up

to 90% of this pectin added as a stabilizer to AMDs is not di-

rectly interacting with the milk gel particles (largely consisting

of casein micelles). This nonadsorbing fraction of pectin, which

is, however, necessary to produce a stable AMD can be taken

out of the stable system without loss of stability. Another way

to decrease the amount of ineffective pectin in the AMD is to

increase the energy input during the homogenization step, that

is, after the addition of pectin. This will increase the adsorption

rate of the pectin, and therefore allows for a smaller overall

pectin concentration.

The steps in the manufacture of fermented dairy beverages

include selection of a milk base with low total solids and low

proteins (Rossi and Clementi, 1983; Dellaglo, 1984), selection

of additives such as fruit juice concentrate, colors, artificial

sweetener, sugar stabilizers, etc. taking into consideration their

interactions with fermented milk components (Rasic and Kur-

mann, 1988), Low temperature (e.g.,


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1% lactic acid, but for therapeutic purposes 0.6–0.7% is more

common.

Another variation has been the introduction of a sweet aci-

dophilus milk, one in which the LA culture has been added but

there has been no incubation. It is thought that the culture will

reach the gastro intestinal (GI) tract where its therapeutic ef-

fects will be realized, but the milk has no fermented qualities,thus delivering the benefits without the high acidity and flavor,

considered undesirable by some people.

Sour Cream

Cultured cream usually has a fat content between 12–30%,

depending on the required properties. The starter is similar to

that used for cultured buttermilk. The cream after standardiza-

tion is usually heated to 75–80◦C and is homogenized at >13

MPa to improve the texture. Inoculation and fermentation con-

ditions are also similar to those for cultured buttermilk, but thefermentation is stopped at an acidity of 0.6%.

Others

There are a great many other fermented dairy products, in-

cluding Kefir, Kumiss, beverages based on bulgaricus or bifidus

strains, labneh, and a host of others. Many of these have devel-

oped in regional areas and, depending on the starter organisms

used, have various flavors, textures, and components from the

fermentation process, such as gas or ethanol.

Fermented Dietetic Preparations

Dietetic foods are manufactured to meet particular nutri-

tional needs of people whose normal processes of assimilation

or metabolism are changed, or for whom a particular effect is

obtained by a controlled intake of food or certain nutrients.

They may be formulated for people suffering from physiologi-

cal disorders or for healthy people with additional needs (Ben-

der, 1975). The manufacture of dieteic-fermented milks causes

problems with texture (correction with ropy starter cultures and

hydrocolloids) and with flavor (correction with mild acidifying

and aroma forming cultures, lactose- hydrolysis, skim-milk con-

centrate, fruit concentrates, sweeteners, etc.). Fermented milk

products fortified with some dietary components (fiber, vita-

mins, phospholipids, minerals, other bioactive substances). At

present, the most frequently produced fermented dietetic prod-

ucts are calorie reduced and dietary fiber fortified (separated and

combined ones). A small number of dietetic products have been

developed for adults (obesity, sports activity, etc.) and for el-

derly people. Only a few of the presently manufactured dietetic

preparations are fermented, a factor which certainly become of

more significance in the future. The use of selected intestinal

bacteria starter cultures for the preparation of fermented milk

products will be an expanding product area. Sarkar and Misr

(2001) attempted to incorporate Bifidobacterium bifidum NDR

and propionibacterium freudenreichii subsp. Shermanii MTC

1371 along with Yoghurt- YH-3 to obtain dietetic yoghurt wit

enhanced nutritional and therapeutic characteristics. Dieteic yo

ghurt was obtained from skim milk (0.5% fat, 7.64% SNF

heated to 95◦C for 30 min at 1% level each and incubated a42◦C. The dietetic yoghurt possess desirable technological an

dietetic characteristics including antibacterial activity again

pathogenic test organisms. The viable cell populations obtaine

in the product were within the range required for successfu

implantation in the intestine.

FUNCTIONAL ASPECTS OF FERMENTED

MILK PRODUCTS

Fermented Products as Vehicles for Fortification

Fortification of foods can be an efficient tool to combat m

cronutrient deficiencies. The successful application of food fo

tification technology is based on consideration of compatibilit

of vehicle, fortificant, and process. Modified foods includin

those that have been fortified with nutrients also fall within th

realm of functional foods (ADA reports). According to Ame

ican Dietetic Association (ADA) functional foods includin

whole foods, fortified enriched or enhanced foods have poten

tially beneficial effect on the health when consumed as apar

of varied diet on a regular basis at effective levels. Fermente

dairy products, such as yoghurt and soft cheeses, have bee

used in iron fortification programmes (Lysionek et al., 2002

In the iron fortification of powdered nonfat dry milk, ferrousulfate at a level of 10 ppm was found to be stable for a perio

of 12 months. Ferric ammonium citrate and ferric chloride at

level of 20-ppm iron in the reconstituted product gave accep

able results (Coccodrilli and Shah, 1985). Enrichment has bee

used interchangeably with fortification (FAO/WHO, 1994), bu

elsewhere it has been defined as the restoration of vitamins an

minerals lost during processing (Hoffpauer and Wright, 1994

Probiotic Applications

The claimed beneficial effects from the consumption of fer

mented milks were once a debated issue. Research conducte

since the turn of the century has however, enhanced the under

standing of the therapeutic effects resulting from the consump

tion of these products. The probiotic products are now consid

ered helpful in maintaining good health, restoring body vigo

and in combating intestinal and other disease disorders (Mita

and Garg, 1992). Most scientific papers refer to research usin

L. acidophilus and bifidobacterium species as dietary culture

Nearly all probiotics currently on the market contain Lacto

bacilli, Streptococci, Enterococci, or Bifidobacteria. In contra

to Japan, where freeze-dried microorganisms are consumed b


12/16


a substantial part of the human population, in Europe, probi-

otic action toward humans is only claimed for certain fermented

dairy products (e.g., yoghurts). Those species that have been

extensively studied so far, with several experimental inocula-

tion of milk for yoghurt manufacture (Prevost & Divies, 1988).

Additional benefits of microencapsulation of cells include: pro-

tection of cells inside the beads from bacteriophages (Steen-son et al., 1987); increased survival during freeze drying and

freezing (Kearney et al., 1990; Sheu and Marshall, 1993; Sung,

1997) The trials on man were mainly carried out with the two

yoghurt bacteria Streptococcus thermophilus and Lactobacillus

bulgaricus. L. casei, Bifidobacteria, and L. acidophilus has also

received important scientific interest, however, only a few hu-

man studies have been reported. From the technological point

of view a good probiotic should be stable and viable for long

periods under storage, should be able to survive the low pH

levels of the stomach, be able to colonize the epithelium of the

gastrointestinal tract of the host, should not be pathogenic and,

last but not least, must be capable of exerting a growth promot-

ing effect or a resistance to infectious diseases (Kalantzopoulos,

1997). Protection of probiotics by microencapsulation in hydro-

colloid beads has been investigated for improving their viability

in food products and the intestinal tract (Rao et al., 1989). This

has been proposed for various dairy fermentations (Champagneet al., 1992a, 1992b, 1994) such as fermentation of whey (Audet

et al., 1989) and continuous Kim and Yoon, 1995); and greater

stability during storage (Kim et al., 1988; Reuter, 1990; Kebary

et al., 1998). Many health benefits have been attributed to fer-

mented dairy products and probiotic microorganisms (Salminen

et al., 1998).

The therapeutic significance of probiotic organisms is sum-

marized in Table 3. In order to exert positive health effects, it

is generally assumed that the microorganisms need to be vi-

able. Therefore, the International Dairy Federation (1997) has

Table 3 Fermented products and therapeutic significance

Clinical condition/symptoms Health benefit of fermented products Reference

Lactose maldigestion • Viable yoghurt well tolerated by lactose-deficient

subjects, L.acidophilus aids lactose digestion.

Morley, 1979; Gilliland, 1989; Kim and Gilliland, 1983

• Manifested by the presence of breath

hydrogen derived from fermentation

of the lactose in the large intestine,

abdominal pain, meteorism, bloating,

flatulence and diarrhea.

• The presence of bacterial b-galactosidase in the

viable yoghurt culture.

Hepatic encephalopathy

• A neurological disorder where

Detoxification of ammonia produced

in the intestine is impaired in patients

with liver failure.

• Alteration of the intestinal microflora Macbeth et al., 1965; Kavasnikov and Sodenko, 1967;

Read et al., 1968 Loguerolo et al., 1987• A decrease in faecal urease, a lowering of blood

ammonia and clinical improvement when treated

individually with L. acidophilus, Lactobacillus

GG, and E. faecum. SF68

Side effects of radiotherapy

• Radiotherapy alters the intestinal

microflora, vascular permeability of

the mucosa and intestinal motility.

• Treatment with fermented milk containing NCFB

1748 significantly decrease pelvic radiotherapy

associated diarrhea

Salminen et al., 1998 ; Kansal, 2001

Tumor or defective immune systems • Lactobacilli and their metabolic products modify

both the immune responses and antitumor

activities.

Kansal, 2001

• Enhanced mainly by activating macrophage

functions and increasing the activity of natural

killer cells and T-cells, maintenance of normal

intestinal flora

Varnam and Sutherland (1994). Anand et al., 1985

• Help removal of dietary procarcinogens. Benno et al., 1984

High serum cholesterol levels • L.acidophilus strains and some bifidobacteria

species lower cholesterol

Mann and Spoerry, 1974; Kawamura et al., 1981;

Gilliland et al., 1985; Gilliland, 1989; Walker and

Gilliland, 1993; Hosona and Tono-oka, 1995; Gopalet al., 1996; Marshall, 1996; Usman and Hosono,

2000; Gupta and Prabhu (2004)

Renal mal function • Effect of fermented milk product on cholesterol

metabolizing enzymes systems in liver

• Promotion of excretion of cholesterol through

faeces,

• Inhibition of cholesterol adsorption by the binding

of cholesterol to cells of LAB.

• The promotion of excretion by binding of bile salts

to lactic acid bacterial cells.

• Bifidobacterium spp.; Lactobacillus acidophilus

• Reduce level of toxic amines.


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defined fermented milk as a milk product fermented by the ac-

tion of specific microorganisms and resulting in reduction of

pH and coagulation. These specific microorganisms shall be

viable, active, and abundant (at least 107 cfu / g) in the prod-

uct to the date of minimum durability. Probiotics are defined as

live microbial food supplements which beneficially influence the

host by improving its intestinal microbial balance (Fuller, 1989;Ziemer and Gibson, 1998). Both definitions emphasize the vi-

ability of microorganisms. Whether viable microorganisms are

necessary for specific health benefits remains unclear. Health

benefits discussed are: alleviation of lactose maldigestion, ef-

fects on diarrhoea, immune-stimulation, antitumor activity, an-

timutagenic activity, reduction of serum cholesterol, and effects

on candidiasis. Many of the studies discussed here were not

conducted for the purpose of determining the efficacy of non-

viable bacteria or fermented milks, but killed microorganisms

have often been used as placebo. The results from these studies

give an indication of the activity of viable versus nonviable mi-

croorganisms and assist in defining the need for probiotics to be

viable.

Fermented Milk Products With Selected Strains

of Intestinal Bacteria

Fermented products containing pure strains of LA, bifidofac-

teria, or either of these in combination with other LAB fall in

the group of probiotic foods and beverages. Important progress

has been made in the study of gut bacteria in relation to their

distribution, metabolic activities, and significance in health and

disease. The potential beneficial roles of indigenous lactobacilliand bifidobacteria are described in many publications (Sandine

et al., 1972; Poupard et al., 1973; Speck, 1976; Shahani and

Chandan, 1979; Rasic and Kurmann, 1983; Goldin and Gor-

bach, 1984) The technology of some of these products has been

given by Rasic and Kurmann (1988). A list of such products and

the organisms associated with them is shown in Table 3. The

concept of yoghurt as aprobiotic carrier food has been reviewed

by Lourens-Hattingh et al., 2001.

Ishibashi and Shimamura (1993) reported that the fermented

milks and LAB beverages association of Japan has developed a

standard which requires a minimum of 107 viable bifidobacteria

cells/ml to be present in fresh dairy products. The criteria de-

veloped by National Yoghurt Association (NYA) of the UnitedStates specifies 108 cfu/g of LAB at the time of manufacture as a

prerequisite to use the NYA “Live and Active Culture” Logo on

the containers of the products (Kailasapathy and Rybka, 1997).

Several publications indicate that fermented milks have antimi-

crobial properties (Jay, 1982), antitumor activities (Reddy et al.,

1983; Shahani et al., 1983) and antileukemic activity (Esser

et al., 1983) The therapeutic significance of fermented prod-

ucts are given in Table 3. The potential therapeutic benefits of

health promoting fermented dairy products are briefly updated

by Itsaranuwat et al. (2003).

Future Research Prospects

The development of more probiotic products by the incor

poration of probiotic organisms into the manufacturing proces

for well-accepted traditional dairy products is to be explore

significantly. The development of convenience based dry cu

tured products containing viable bacteria by various technique

such as microencapsulation and the fortification of such powder

or concentrated products with minerals and vitamins are gain

ing importance. The preparation of suitable blends containin

healthful ingredients such as dietary fiber, natural vitamins an

folic acid also offers new possibilities for this widely expandin

research area. The formulation of various health drinks and d

etetic preparations as value added products based on traditiona

fermented products will definitely bring more opportunities t

the beverage industry. There is a need to intensify research t

distinguish the therapeutic benefits of viable and nonviable or

ganisms in different fermented products apart from the genera

nutrient enhancement achieved.

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