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Biotechnological innovations in
kefir production: a review

Innovations in
kefir production

S. Sarkar
Metro Dairy Limited, Neelgunj Bazar, West Bengal, India

283

Abstract
Purpose The purpose of the paper is to show that traditionally, kefir was obtained by fermenting
milk with kefir grains. Wide variation in microflora of kefir grains makes it difficult to obtain an
optimal and uniform starter culture necessary for obtaining a quality kefir. Reviewed literature on
microbiological and technological innovations in kefir production would enrich the scientific
knowledge resulting in production of kefir with superior physical, chemical, nutritional, therapeutic
and sanitary qualities.
Design/methodology/approach An attempt is made to highlight the microbiological and
technological aspects of kefir production with regard to the microflora of kefir grains, suitability of
different types of milk, treatment of milk, starter inoculation and incubation, packaging, storage and
post-production treatment of kefir as well as methods of preservation of kefir grains.
Findings Diverse microflora of kefir grains is the prime cause for the wide variation in kefir
quality. Production of kefir is based on symbiotic relation between lactic acid bacteria and yeasts and
the type of milk, their heat-treatment, size of inoculating starters and temperature of incubation
influence their metabolic activities. Application of a suitable combination of lactic acid bacteria and
yeasts would enable production of kefir with more uniform product with specific properties Packaging
of kefir in a suitable container and storage at low temperature are suggested to retain its qualities.
Originality/value Fermentation of milk with a suitable starter combination consisting of lactic
acid bacteria and yeasts rather than application of kefir grains during the production of kefir would be
more scientific to yield a product with enhanced nutritional and therapeutic qualities.
Keywords Heat treatment, Milk, Food products, Drinks
Paper type Research paper

Introduction
Yeasts are commercially significant in food industries, particularly in dairy processing,
because they bring about desirable fermentative changes in some of the fermented milk
products (Westall and Filtenborg, 1998) and have remarkably stable association with
lactic acid bacteria (LAB), especially lactobacilli (Wood, 1981). kefir is a
self-carbonated, sour beverage obtained with a fermenting agent called kefir grain,
consisting of casein and gelatinous colonies of LAB and yeasts grown together
symbiotically (Webb et al., 1987). Yeasts and lactobacilli are mutually dependent and
grow in balanced proportion in kefir grains (Wood and Hodge, 1985) and symbiosis
between yeasts, lactobacilli and streptococci was noted during the production of kefir
(Loretan et al., 2003).
Kefir has gained popularity in various part of the world including Southwest Asia,
Eastern and Northern Europe, North America, Japan (Otles and Cagindi, 2003), Middle
East, North Africa and Russia (Koroleva, 1982, IDF, 1988) owing to its nutritional
(Guzel-Seydim et al., 2003, Liut Kevicius and Sarkinas, 2004) and therapeutic properties
(Marquina et al., 2002, Liu et al., 2002, Zacconi et al., 2003, Czamanski et al., 2004) and
has been recommended for consumption as a dietetic beverage (Sarkar, 2007). Wide
variation in microflora of kefir grains makes it difficult to obtain an optimal and

British Food Journal

Vol. 110 No. 3, 2008
pp. 283-295
q Emerald Group Publishing Limited
0007-070X
DOI 10.1108/00070700810858691

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uniform starter culture necessary for obtaining a quality kefir beverage. Factors
affecting kefir flora and consequently the quality of kefir include media used, grain to
milk ratio, cultures used for kefir production, time and temperature of incubation,
extent of agitation, type of package and storage conditions (Koroleva, 1988a, 1991).
According to the Codex Standard, composition of kefir must be constituted of , 10.0
per cent (m/m) milk fat, a minimum of 2.80 per cent (w/w) milk protein, 0.6 per cent
(m/m lactic acid) titratable acidity, 107cfu/g total microbial count and 104cfu/g yeast,
however no specification for ethanol content has been mentioned (FAO/WHO, 2001). In
the present endeavor biotechnological innovations for production of kefir with
improved dietetic properties have been reviewed.
Production of kefir
Kefir was first made from goat milk with kefir grains in goatskin bag by hanging in the
house during winter and outside during summer (Ozer and Ozer, 1999). Traditionally,
kefir was produced by inoculating milk with kefir grains (Pijanowski, 1980) or by the
widely adopted European method, which involved use of bulk milk culture obtained by
kefir grains for milk inoculation (Puhan and Vogt, 1985). Kurmann (1984) classified kefir
under the class of mixed lactic acid and ethanol fermented milk and can be further
sub-classified as kefir obtained using kefir grains and artificial kefir obtained without
kefir grains Various steps involved during kefir production are enunciated below.
Microflora of kefir grains
Kefir grains are sometimes called Millet of the prophet or Mohomet grains, dump
shaped, gelatinous granules measuring about 1-2 mm to 3-6 mm, sometimes up to
2-15 mm in diameter with irregular, rough and convoluted surfaces (Koroleva, 1991). It
has been reported that kefir grains are constituted of 108-109cfu/ml LAB, 105-106cfu/ ml
yeasts and 105-106cfu/ml acetic acid bacteria (Koroleva, 1991; Garrote et al., 2001). Basic
microflora of kefir grain consists of LAB such as lactobacilli (thermophilic and
mesophilic), leuconostoc, streptococci (homofermentative and heterofermentative),
lactococci and acetic acid bacteria as well as yeasts (Bottazzi et al., 1994, Rea et al.,
1996), all held together by water soluble polysaccharide called kefiran (Kosikowski,
1977), which is composed of glucose and galactose (Yokoi et al., 1991). Bosch et al. (2006)
identified Lactobacillus kefir, Lactobacillus parakefir and Lactobacillus brevis
(heterofermentative) and Lactobacillus plantarum, Lactobacillus acidophilus,
Lactobacillus kefirgranum, Lactobacillus kefiranofaciens and Lactobacillus casei
(homofermentative) from kefir grains. Ninane et al. (2005) reported greater variability
in the population of lactic acid streptococci (443 per cent) than lactobacilli (28 per cent)
and yeasts (35 per cent), isolated from kefir grains. Witthuhm et al. (2004) reported that
LAB and yeasts present in kefir grains vary widely and counts ranged from 6.4 104
8.5 108 and 1.5 105 3.7 108 cfu/ml, respectively. Identified LAB and yeasts in
kefir grains and kefir are shown in Tables I and II. Duitschaever et al. (1988) observed the
presence of short and elongated bacilli as well as yeasts embedded amongst a densely
packed fibrillar, amorphous matrix but no cocci in kefir grains. They further reported
appearance of some long and curved bacteria in the surface section of the grain
indicating disparity in the microflora at the edge and near the centre of kefir grains.
Dominance of outer layer of kefir grain with rod shaped LAB, yeasts at the core, a
balanced bacteria and yeast at the intermediate zone and a progressive change

Constituents of bacterial flora

Lactobacilli
Lactobacillus kefir
Lactobacillus kefiranofaciens
Lactobacillus kefirgranum
Lactobacillus parakefir
Lactobacillus brevis
Lactobacillus plantarum
Lactobacillus paraplantarum
Lactobacillus gasseri
Lactobacillus helveticus
Lactobacillus acidophilus
Lactobacillus delbrueckii
Lactobacillus rhamnosus
Lactobacillus casei
Lactobacillus paracasei
Lactobacillus fructivorans
Lactobacillus hilgardii
Lactobacillus fermentum
Lactobacillus viridescens
Lactobacillus bulgaricus
Lactococci
Lactococcus lactis subsp. lactis
Lactococcus lactis subsp. cremoris
Streptococci
Streptococcus thermophilus
Enterococci
Enterococcus durans
Enterococcus faecium
Leuconostocs
Leuconostoc mesenteroides
Leuconostoc mesenteroides subsp. cremoris
Acetic acid bacteria
Acetobacter aceti
Acetobacter pasteurianus
Other bacteria
Escherichia coli
Bacillus subtilis
Micrococcus sp.

References
Takizawa et al. (1994)
Takizawa et al. (1994)
Takizawa et al. (1994)
Takizawa et al. (1994)
Santos et al. (2003)
Santos et al. (2003)
Anana et al. (2005)
Anana et al. (2005)
Simova et al. (2002)
Santos et al. (2003)
Santos et al. (2003)
Santos et al. (2003)
Simova et al. (2002)
Santos et al. (2003)
Yoshida and Toyoshima (1994)
Yoshida and Toyoshima (1994)
Angulo et al. (1993)
Angulo et al. (1993)
Wang et al. (2004)

Innovations in
kefir production

285

Yoshida and Toyoshima (1994)

Koroleva (1991)
Simova et al. (2002)
Yuksekdag et al. (2004)
Wang et al. (2004)
Koroleva (1991)
Mainville et al. (2006)
Koroleva (1991)
Ottogalli et al. (1973)
Angulo et al. (1993)
Ottogalli et al. (1973)
Angulo et al. (1993)

according to the distance from the core has been reported (Ottogalli et al., 1973; Rea
et al., 1996). Microflora of kefir may vary with the type of culturing media used as well
as the method of kefir production employed. No disparity in microflora and
polysaccharide composition of kefir grains were observed during culturing in milk or
whey (Fil Chakova and Koroleva, 1997), however higher polysaccharide contents were
recorded in cow milk than in soya milk (Abraham and Antoni, 1999). Microflora
detected in kefir was lactobacilli, streptococci and yeasts, when made from these pure
cultures or cocci and few yeast, when made by direct-set culture (Duitschaever et al.,
1988). Recently, scanning electron microscopy and transmission electron microscopy of

Table I.
Bacterial flora of kefir
grains and kefir

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Table II.
Yeast flora of kefir grains
and kefir

Constituents of yeast flora

Klyveromyces species
Klyveromyces matxianus
Klyveromyces lactis
Saccharomyces species
Saccharomyces cerevesiae
Saccharomyces unisporus
Saccharomyces exiguss
Saccharomyces turicensis
Saccharomyces delbrueckii
Torulaspora species
Torulaspora delbrus
Torulaspora delbrueckii
Candida species
Candida pseudotropicalis
Candida tenuis
Candida inconspicua
Candida maris
Candida lambica
Candida tannotelerans
Candida valida 6
Candida kefyr
Candida holmii
Other yeasts
Pichia fermentans
Zygosaccharomyces rouxii
Debaryomyces hansenii
Bretannomyces anomalus
Issatchenkia occidentalis

References
Loretan et al. (2003)
Loretan et al. (2003)
Loretan et al. (2003)
Loretan et al. (2003)
Iwasawa et al. (1982)
Wyder and Puhan (1997)
Rosi (1978)
Loretan et al. (2003)
Loretan et al. (2003)
Ottogalli et al. (1973)
Ottogalli et al. (1973)
Simova et al. (2002)
Simova et al. (2002)
Engel et al. (1986)
Dousset and Caillet (1993)
Dousset and Caillet (1993)
Engel et al. (1986)
Engel et al. (1986)
Angulo et al. (1993)
Loretan et al. (2003)
Loretan et al. (2003)
Wyder and Puhan (1997)
Engel et al. (1986)

kefir grains revealed most denser colonized portion at the exterior surface, consisting
mainly of bacteria and yeasts, many of them autolysing but could not pass through the
polysaccharide matrix (Zhang et al., 1998). Lin et al.(1999) also reported that LAB were
localized mainly on the surface layer consisting of Klyveromyces marxianus and
Leuconostoc mesenteroides whereas yeasts such as Pichia fermentas at the center of the
grains.
Type of milk
Milk intended for fermented milk production should comply with the following
requirements of low bacterial count, absence of pathogens or inhibitory substances
such as antibiotic residues and sanitizer residues (Roginski, 1988). kefir is frequently
made from whole, part skim or skim milk (Webb et al., 1987) employing milk from cow,
ewe, goat, mare (Kneifel and Mayer, 1991), camel (Garg, 1989) or sheep (Wojtowski
et al., 2003). Goat milk is not suitable for kefir manufacture due to lower viscosity and
sensory properties in contrast to cow milk kefir and no significant improvement in the
sensory scores due to softer consistency could be achieved with 2 per cent
supplementation of goat milk kefir with milk powder, whey protein concentrate and
inulin in comparison to un-supplemented kefir (Bozanic et al., 2003). Recently literature
indicated that quality of kefir could be influenced with the acquiring period of milk.

Jasinska et al. (2005) concluded that best quality kefir with good consistency, highest
acidification and least perceptible goaty flavour could be obtained by utilizing goat
milk collected at the end of October. Amongst milk from different breeds (cow, goat
and sheep), sheep milk should be preferred for kefir production due to certain health
benefits owing to its lowest contents of medium-chain saturated acids and highest
linoleic and / -linolenic acid (Wojtowski et al., 2003). Recently, production of kefir
from soy milk has also been reported (Halle et al., 1994; Kuo and Lin, 1999).
Treatment of milk
Concentration of skim milk to . 1.3 to 1.8 fold adopting ultrafiltration technique is
suggested for obtaining denser kefir (Chagarovsky and Lipatov, 1990). Milk intended
for cultured milk production must be heat-treated with the objective of pasteurizing the
product, rendering milk more stimulatory growth medium for starter cultures due to
elaboration of amino acids and other growth factors, reduction of the redox potential
and elimination of inhibitory substances, improve physical properties of fermented
milk and to reduce syneresis and prevent hydrolytic rancidity through inactivation of
enzyme lipase (Kessler, 1981). Various temperature-time combinations have been
suggested for heat-treatment of milk during kefir production. Suggested
heat-treatments were 858C/30 min (Vedamuthu, 1977), 85-908C/15-20 min (Klyavinya,
1980), 90-938C/15 min (Penido et al., 2001), 928C/20 min (Simova et al., 2006),
958C/15 min (Ozer and Ozer, 1999), 90-958C/2-3 min (Koroleva, 1988b) and
958C/10-15 min (Dies, 2000).
Starter inoculation and incubation
Kefir can be obtained either by inoculating milk with kefir grains or by inoculating
milk with bulk starter obtained by culturing with kefir grains. Beshkova et al. (2002)
produced kefir involving a starter comprising of two bacteria (Lactobacillus helveticus
and Lactobacillus lactis subsp. lactis), single yeast culture isolated from kefir grains
(Saccharomyces cerevesiae) along with two yoghurt strains (Lactobacillus delbrueckii
subsp. bulgaricus and Streptococcus thermophilus). In USA, kefir is produced
employing only starter cultures consisting of Streptococcus lactis, Lactobacillus
plantarum, Streptococcus cremoris, Lactobacillus casei, Streptococcus diacetylactis,
Leuconostoc cremoris and Saccharomyces florentinus (Hertzler and Clancy, 2003).
Factors influencing extent of acidification during kefir production are size of
inoculation of kefir grains, agitation and temperature of incubation (Irigoyen et al.,
2003). Extent of acidification and carbondioxide production was influenced by the
concentration of kefir grains, higher being noted at a level of 100 g than 10 g/l milk
(Garrote et al., 1998). kefir production with 5 per cent kefir grains proved to be optimum
for ethanol and volatile acid production (Korovkena et al., 1978). Rate of inoculation
also influence the kefir flora. kefir made with 1 per cent inoculation had LAB as the
dominant flora but those made with 5 per cent inoculation were predominated with
yeasts and acetic acid bacteria (Irigoyen et al., 2005).
The fermentation temperature affects distinctly both the total capacity of
acidification and the acid production speed (Irigoyen et al., 2003). Chagarovsky and
Zholkevskaya (2003) reported that acid production occurred within 6 h at 408C and
308C for thermophilic and mesophilic bacteria, respectively. Mesophilic microflora
considerably increased during the first 24 h of fermentation and then remained stable

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with a slight increase at the end of kefir production (Penido et al., 2001). During the
same period of fermentation, the viable population of bacteria (lactobacilli and
lactococci), yeasts and acetic acid bacteria attain a level of 108, 105 and 106cfu/ml,
respectively (Irigoyen et al., 2005). Garcia-Fortan et al. (2006) noted predominance of
Lactococcus sp. during the first 48 h of fermentation (< 8.0 log cfu/g), followed by
predominance of Lactobacillus sp. (< 8.5 log cfu/g). Liu et al. (2002) recorded that
immediately after inoculation of soya milk with kefir grains, counts of LAB was higher
than yeasts and their activity enhanced with fortification of soya milk with 1 per cent
glucose.
During fermentation various flavouring compounds are produced by starter
cultures. Acetaldehyde is produced by L. delbrueckii subsp. bulgaricus, diacetyl by S.
thermophilus, L. helveticus and L. lactis subsp. lactis, acetone by L. delbrueckii subsp.
bulgaricus and L. helveticus, ethanol by S. cerevesiae (Beshkova et al., 2002). Garg (1989)
mentioned other flavouring compounds traced in kefir to be propionaldehyde,
2-butanone, isoamyl alcohol and diacetyl. Acceptability of kefir produced by pure
cultures could be enhanced either by sweetening (Duitschaever et al., 1987, 1991),
addition of peach flavour or modification of fermentation process with the addition of
lactococci, lactobacilli or yeasts (Duitschaever et al., 1991; Muir et al., 1999).
Recommended incubation temperature-time combinations are 208C/20 h 11 h (Chen
et al., 2006), 208C/48 h (Abraham and Antoni, 1999), 20-238C/12-14 h (Pijanowski, 1980),
228C/11 h (Li et al., 2004), 22-238C/20 h (Bottazzi, 1985), 24-278C/20 h (Klupsch, 1985) and
22-258C/8-12 h (Koroleva, 1988b) for kefir production. Recently, Simova et al. (2006)
recommended two-stage fermentation process (288C/5 h and 208C/16 h) for kefir
production.
Packaging
Kessler (1981) mentioned that the containers to be used for packaging of fermented
milk should be impermeable to water and odours, insoluble in water and free from
foreign odours. Kwak et al. (1996) mentioned that growth of yeasts continue after
packaging, therefore the containers intended for kefir packing must be either strong
enough to withstand the buildup pressure such as glass or flexible enough to retain the
amount of gas produced such as plastic with an aluminium foil top. Special containers
designed for kefir had lids consisted of three layers that allows the escape of carbon
dioxide generated by viable yeasts, which prevents swelling and bulging of kefir cups
(Fluckiger, 1986).
Storage
Ozer and Ozer (1999) suggested that post-fermentation cooling of kefir to 4-68C should
be done slowly within 10-12 h to ensure retention of its pronounced aroma and typical
taste. A slight abatement in contents of orotic and citric acid but a significant
enhancement in lactic acid to 7,739ppm, ethanol to 0.08 per cent, acetaldehyde to
11mg/g, decline in acetoin to 16ppm and presence of non-detectable levels of diacetyl
during storage of kefir at 48C/21 days were encountered (Guzel-Seydim et al., 2000).
Irigoyen et al. (2005) reported a decline in lactic flora (1.5 log units) but a stability for
acetic acid bacteria and yeasts during storage of kefir at 5 ^ 18C/28 days. Storage of
kefir for two days prior to its consumption is recommended for better sensory qualities
and highest viable population of Lactobacillus gasseri (Anana et al., 2005).

Post-production treatments
Shelf-life of kefir is dependent on the type of packaging material and varies from eight
to ten days at 3-48C (Koroleva, 1988b). Post-production treatments such as autoclaving,
irradiation, ohmic heating and high pressure treatment can be done on kefir (Mainville
et al., 2001) with the objective of enhancing the shelf-life. High pressure treatment of
kefir at 400 MPa/30 minutes induced deactivation of bacteria and yeasts (Mainville
et al., 2001) and loss of ability of LAB to inhibit test organisms (Jankowska et al., 2001)
but no change in protein and lipid structure (Mainville et al., 2001). Since loss of
viability and antibacterial activity of starter cultures will reduce the therapeutic value
of kefir, application of high-pressure treatment for kefir intended for therapeutic
application is not recommended. Freeze drying of kefir also induced a loss in viability
of microorganisms but the survival can be improved with the addition of 10%
galactose or 10% sucrose prior to freeze drying (Chen et al., 2006).
Biotechnological innovations in kefir production
Traditional kefir made from goat milk had low viscosity and sensory properties in
contrast to cow milk kefir and contained 0.04-0.3 ml/100 ml ethanol (Seiler, 2003).
Tratnik et al. (2006) recommended supplementation of goat milk with whey protein
concentrate at a level of 60.0-60.5 g/100 g proteins for enhancement in ethanol
production (0.35 ml/100 ml). To comply with the consumers demand for more healthy
foods, soya milk could be a suitable substitute for kefir production owing to its low
saturated fat and cholesterol (Berry, 2000), higher polyunsaturated fatty acids, lecithin,
linolenic acid, magnesium, iron, folic acid and vitamin E (Hermann, 1991). Koca et al.
(2002) reported that though soya milk is a satisfactory medium for the growth and
activity of starter cultures, a slower rate of acidification is noted in soya milk in
contrast to those in cow milk Fortification of soya milk with glucose (Liu and Lin, 2000)
and low-fat milk with tryptose (Schoevers and Britz, 2003) are suggested for growth
stimulation LAB and yeasts.
Fermentation of milk by traditional methods employing kefir grains resulted in
disparity in product quality due to diverse microflora and uncontrolled fermentation.
Two methods have been suggested to overcome the drawbacks of traditional methods
of kefir production kefir can be produced either by simultaneous (Tamai et al., 1996) or
consecutive lactic acid and yeast fermentation (Beshkova et al., 2002)
Production of kefir employing pure cultures induced production of higher
concentration of carbonyl compounds (Beshkova et al., 2003) and carbondioxide (1.7 vs
0.85 to 1.05 g/L) in contrast to those obtained adopting kefir grains (Gobbetti et al.,
1990, Beshkova et al., 2002; Simova et al., 2002). Klupsch (1985) recommended
primarily fermentation of milk with a 2 per cent starter comprising of streptococcus
and lactococci, followed by a secondary fermentation with 0.05-0.5 per cent starter
containing Candida kefir and Lactobacillus brevis. Best quality kefir could be produced
by incubating homogenized milk with 3 per cent (V/V) each of bacteria and yeast at
258C for 24 h (Assadi et al., 2000).
Probiotic kefir capable of exhibiting antimicrobial properties could be obtained
employing L. acidophilus, Lactobacillus kefiranofuctens and Lactobacillus
kefiranofaciens (Santos et al., 2003) or Bifidobacterium bifidum (Murashova et al.,
1997). Introduction of Candida kefyr during kefir production may be advantageous as it
did not disappeared completely at pH 2.0, retains 97.2 per cent viability in presence of 1

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per cent bile salts and not inhibited by most antibiotics including tetracycline (You
et al., 2006). During kefir production introduction of 3 per cent L. delbrueckii subsp.
bulgaricus HP 1 with S. thermophilus T 15 (1:1), 1 per cent L. lactis subsp. lactis C 15, 3
per cent L. helveticus MP 12 (Simova et al., 2006) have been recommended and further
inclusion of Sacch. cerevesiae AB shortened the time to reach the maximum
exopolysaccharide production (824.3 mg/L) by 6 h.
In order to meet the consumers demand for healthful foods in the current era of
self-care and complementary medicine kefir with enhanced dietetic properties could be
obtained by co-inoculation of soya milk with yeasts (Sacch. cerevesiae, Candida kefir),
lactic acid bacteria (Streptococcus thermophilus, Lactobacillus bulgaricus) and probiotic
cultures (Lactobacillus acidophilus, Bifidobacterium bifidum).
Preservation of kefir grains
During fermentation, kefir grains comes at the surface due to production of carbon
dioxide and are strained, washed, dried and stored for longer periods between their
application. Liu et al. (1999) recommended storage of lyphilized kefir grains at 2 208C
and viability of LAB retained stable and resulted in kefir with viable population
similar to those obtained from non-stored grains. kefir grains could be preserved by
frozen storage without milk at 2 188C, refrigerated storage at 48C, air drying at room
temperature for three weeks in a dessicator or freeze drying but air drying is not
recommended for commercial application due to unacceptable colour and flavour of
grain (Witthuhm et al., 2005a). Among various packaging materials low-density
polyethylene film (LPDE), oriented polyster film (OPET) and methallized oriented
polyster film (MOPET), adoption of MOPET is recommended for retaining the activity
of kefir grains for extended storage period (Witthuhm et al., 2005b). Selection of proper
packaging material and preservation technique is of utmost imporancy for longer
period of storage of kefir grains.
Conclusion
Traditional method of kefir production involved unpredictable and slow souring of
milk using kefir grains, resulting in disparity in quality of kefir produced.
Biotechnological innovations must be considered to upgrade the dietetic characteristics
of kefir There is exigency to screen out the method for enhancing the shelf life of kefir
to extend its market reach. Every effort should be exercised to popularize kefir as a
dietetic beverage.
References
Abraham, A.G. and Antoni, G.L.D. (1999), Characterization of kefir grains in cows milk and in
soya milk, J. Dairy Res., Vol. 66, pp. 327-33.
Anana, I., Ortigosa, M., Irigoyen, A., Ibanez, F.C. and Torre, P. (2005), Isolation and
identification of potentially probiotic lactobacillus strains from kefir, Milchwiss., Vol. 60,
pp. 292-4.
Angulo, L., Lopez, E. and Lema, C. (1993), Microflora present in kefir grains of the Galician
region (North-West of Spain), J. Dairy Res., Vol. 60, pp. 263-7.
Assadi, M.M., Pourahmad, R. and Moazami, N. (2000), Use of isolated kefir cultures in kefir
production, World J. Microbiol. Biotechnol., Vol. 16, pp. 541-3.
Berry, D. (2000), Bean vs bovine, Dairy Fd., Vol. 101, pp. 37-9.

Beshkova, D.M., Simova, E.D., Frengova, G.I., Simov, Z.I. and Dimitrov, Z.P. (2003), Production
of volatile aroma compounds by kefir starter cultures, Int. Dairy J., Vol. 13, pp. 529-35.
Beshkova, D.M., Simova, E.D., Simov, Z.I., Frengova, G.I. and Spasov, Z.N. (2002), Pure cultures
for making kefir, Fd. Microbiol., Vol. 19, pp. 537-44.
Bosch, A., Golowczyc, M.A., Abraham, A.G., Garrote, G.L., Antoni, G.L.D. and Yantorno, O.
(2006), Rapid discrimination of lactobacilli isolated from kefir grains by FT-IR
spectroscopy, Int. J. Fd. Microbiol., Vol. 111, pp. 280-7.
Bottazzi, V. (1985) in Rehm, H.L. and Reed, G. (Eds), FRG, Vol. 5, Verlag Chemie, Weinheim,
pp. 315-66.
Bottazzi, V., Zacconi, C., Sarra, P.G., Dallavalle, P. and Parisi, M.G. (1994), kefir microbiology,
chemistry and technology, Industria del Latte, Vol. 30, pp. 41-62.
Bozanic, R., Tratnik, L., Herceg, Z. and Hruskar, M. (2003), The Quality and Acceptability of Plain
and Supplemented Goats and Cows Fermented Milk with Kefir Culture the Quality of
Goats and Cows Kefir, International Dairy Federation, Brussels, pp. 267-79.
Chagarovsky, A.P. and Lipatov, N.N. (1990), Ultrafiltration in the Manufacture of kefir, Quarg
and Domashny Fresh Cheese, International Dairy Federation, Brussels, p. 440.
Chagarovsky, V.P. and Zholkevskaya, I.G. (2003), Biotechnology of Bioyoghurt and biokefir
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Corresponding author
S. Sarkar can be contacted at: metrocal@cal3.vsnl.net.in

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