functional cultures and health benefits.pdf
TRANSCRIPT
-
International Dairy Journal 17
Review
Keywords: L. acidophilus; Bifidobacterium; L. casei; Health benets; Stability; Bioactive peptides
3.6. Reduction in serum cholesterol . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1266
3.7. Helicobacter pylori infection. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1266
1. Introduction
ARTICLE IN PRESSAt the beginning of this century, Nobel Laureate ElieMetchnikoff, at the Pasteur Institute, linked health and
0958-6946/$ - see front matter r 2007 Elsevier Ltd. All rights reserved.
doi:10.1016/j.idairyj.2007.01.014
Tel.: +61 3 9919 8289; fax: +61 3 9919 8284.E-mail address: [email protected]. Inammatory bowel disease . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1266
3.9. Immune system stimulation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1267
4. Prebiotics and synbiotics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1268
5. Alternate products for incorporating probiotics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1268
5.1. Products based on bioactive peptides released from milk proteins by proteolytic probiotics . . . . . . . . . . . . . . . . . . 1271
6. Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1273
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1273Contents
1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1262
2. Probiotic bacteria . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1263
3. Health benets of functional probiotic cultures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1264
3.1. Antimicrobial activity and gastrointestinal infections . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1264
3.2. Effectiveness against diarrhoea . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1265
3.3. Improvement in lactose metabolism . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1265
3.4. Antimutagenic properties . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1265
3.5. Anticarcinogenic properties . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1266probiotics.
r 2007 Elsevier Ltd. All rights reserved.Functional cultures and health benets
Nagendra P. Shah
School of Molecular Sciences, Victoria University, P.O. Box 14428, Melbourne, Vic. 8001, Australia
Received 25 September 2006; accepted 22 January 2007
Abstract
A number of health benets have been claimed for probiotic bacteria such as Lactobacillus acidophilus, Bifidobacterium spp., and
L. casei. These benets include antimutagenic effects, anticarcinogenic properties, improvement in lactose metabolism, reduction in
serum cholesterol, and immune system stimulation. Because of the potential health benets, these organisms are increasingly being
incorporated into dairy foods, particularly yoghurt. In addition to yoghurt, fermented functional foods with health benets based on
bioactive peptides released by probiotic organisms, including Evoluss and Calpiss, have been introduced in the market. To maximize
effectiveness of bidus products, prebiotics are used in probiotic foods. Synbiotics are products that contain both prebiotics and(2007) 12621277
www.elsevier.com/locate/idairyj
-
1900s that scientists began to investigate the reasons forthose benets. A number of genera of bacteria (and yeast)are used as probiotics, including Lactobacillus, Leuconos-toc, Pediococcus, Bifidobacterium, and Enterococcus, butthe main species believed to have probiotic characteristicsare L. acidophilus, Bifidobacterium spp., and L. casei.Members of the genera Lactobacillus and Bifidobacteriumhave a long and safe history in the manufacture of dairyproducts and are also found as a part of gastrointestinalmicroora. Probiotic bacteria with desirable properties andwell-documented clinical effects include L. johnsonii La1,L. rhamnosus GG (ATCC 53103), L. casei Shirota,L. acidophilus NCFB 1478, B. animalis Bb12 and L. reuteri(Shah, 2004).At present, 56 species of the genus Lactobacillus have
been recognized (Table 1). L. acidophilus is the mostcommonly suggested organism for dietary use. Growth ofL. acidophilus occurs at as high as 45 1C, however, theoptimum growth temperature is between 3540 1C. Theorganisms grow in slightly acidic media at pH of 6.44.5,but growth ceases when a pH of 4.03.6 is reached. Theacid tolerance of the organisms varies from 0.3% to 1.9%
ARTICLE IN PRESSiry Journal 17 (2007) 12621277 1263longevity to ingestion of bacteria present in yoghurt(monograph reprinted in: Metchnikoff, 2004). It wasbelieved that bacteria present in yoghurt controlledinfections caused by enteric pathogens and regulatetoxaemia, both of which play a major role in ageing andmortality. This observation provided a major boost to themanufacture and consumption of yoghurt. The healthbenets derived by the consumption of foods containingLactobacillus acidophilus, Bifidobacterium and L. casei arenow well documented.Functional foods are dened as foods that contain some
health-promoting component(s) beyond traditional nutri-ents (Shah, 2001). Functional foods are also known asdesigner foods, medicinal foods, nutraceuticals, therapeuticfoods, superfoods, foodiceuticals, and medifoods (Shah,2001). In general, the term refers to a food that has beenmodied in some way to become functional. One way inwhich foods can be modied to become functional is byaddition of probiotics: the word probiotic originated fromGreek meaning for life. Probiotic foods are dened asfood containing live microorganisms believed to activelyenhance health by improving the balance of microora inthe gut (Fuller, 1992). Probiotic yoghurts, for instance,contain probiotic bacteria as health promoting componentsbeyond traditional nutrients.Traditionally, yoghurt is manufactured using Strepto-
coccus thermophilus and L. delbrueckii ssp. bulgaricus asstarter cultures. These organisms are claimed to offer somehealth benets as postulated by Metchnikoff; however,they are not natural inhabitants of the intestine. Therefore,for yoghurt to be considered as a probiotic product,L. acidophilus, Bifidobacterium and L. casei are incorpo-rated as dietary adjuncts. Products such as Yakult containthe L. acidophilus Shirota strain. Whilst fermented milkproducts containing only one or more of these threeadjuncts could be manufactured, the longer incubationperiod required and poorer resultant product quality arethe two main factors that preclude such practice commer-cially. Thus, the normal practice is to make productwith both starter organisms, e.g., Str. thermophilus andL. delbrueckii ssp. bulgaricus, and one or more species ofprobiotic bacteria.New fermented products containing L. acidophilus,
Bifidobacterium spp., L. casei Shirota, L. rhamnosus GG,and L. reuteri have been developed in Europe. However,L. acidophilus and Bifidobacterium spp. are most commonlyused as probiotics. It is estimated that over 70 productscontaining L. acidophilus and Bifidobacterium spp., includ-ing yogurt, buttermilk, frozen desserts and milk powder,are produced worldwide. Probiotic organisms themselvesare also available as powders, capsules and tablets.
2. Probiotic bacteria
Probiotics have been consumed in foods such as yoghurt
N.P. Shah / International Dafor perhaps thousands of years, and while the cultureswere thought to have benecial effects, it was not until thetitratable acidity, with an optimum pH at 5.56.0 (Curry &Crow, 2003; Shah, 2003).L. acidophilus tends to grow slowly in milk, leading to
the risk of overgrowth of undesirable microorganisms.Ironically, most strains of L. acidophilus do not survive wellin fermented milk due to the low pH, and it is difcult tomaintain large numbers in the product. The poor growth ispartly related to low concentration of small peptides andfree amino acids in milk, which would be insufcient tosupport the bacterial growth.
Table 1
Lactobacilli used as probiotic culturesa
Species Strains
L. acidophilus LA-1/LA-5 (Chr. Hansen)
L. acidophilus NCFM (Rhodia)
L. acidophilus Johsonii La1 (Nestle)
L. acidophilus DDS-1 (Nebraska Cultures)
L. acidophilus SBT-2062 (Snow Brand Milk Products)
L. bulgaricus Lb12
L. lactis L1A (Essum AB)
L. casei Immunitas (Danone)
L. plantarum 299v, Lp01
L. rhamnosus GG (Valio)
L. rhamnosus GR-1 (Urex Biotech)
L. rhamnosus LB21 (Essum AB)
L. reuteri SD2112/MM2 (Biogaia)
L. rhamnosus 271 (Probi AB)
L. plantarum (Probi AB)
L. reuteri SD2112 (also known as MM2)
L. casei Shirota (Yakult)
L. paracasei CRL 431 (Chr. Hnasen)
L. fermentum RC-14 (Urex Biotech)
L. helveticus B02aAdapted from Krishnakumar and Gordon (2001), Holm (2003), Playne
et al. (2003); Shah (2004).
-
Bifidobacterium are normal inhabitants of the humangastrointestinal tract. Recent in vivo scientic studies usinganimals or human volunteers have shown that consump-tion of live Bifidobacterium has an effect on the gutmicroora. Selected strains survive stomach and intestinaltransit and reach the colon in abundant numbers. New-borns are colonized with Bifidobacterium within daysafter birth and the population appears to be relativelystable until advanced age, when a decline in their numbersoccurs. However, diet, antibiotics, and stress are reportedto inuence the population of Bifidobacterium in theintestines.Presently, there are 29 species in the genus Bifidobacter-
ium (Table 2), 14 of which are isolated from human sources(i.e., dental caries, faeces and vagina), 12 from animalintestinal tracts or rumen, and 3 from honeybees.Bifidobacterium species found in humans are: B. adolescen-tis, B. angulatum, B. bifidum, B. breve, B. catenulatum,B. dentium, B. infantis, B. longum, and B. pseudocatenula-tum. B. breve, B. infantis, and B. longum are found inhuman infants; B. adolescentis, and B. longum are found in
ARTICLE IN PRESSN.P. Shah / International Dairy J1264human adults (Shah & Lankaputhra, 2002).The optimum pH for the growth of Bifidobacterium is
6.07.0, with virtually no growth at pH 4.55.0 and belowor at pH 8.08.5 and above. Optimum growth occurs at atemperature of 3741 1C, the minimum and maximumgrowth temperatures are 2528 and 4345 1C, respectively.The main probiotic organisms that are currently used
worldwide belong to the genera Lactobacillus and Bifido-bacterium. A limited number of investigations have alsobeen carried out into the potential properties of generaincluding Pediococcus, Leuconostocs, and Propionibacter-ium and also of Enterococcus faecium. Ent. faecium is
Table 2
Bifidobacterium cultures used as probiotic culturesa
Species Strains
B. adolescentis ATCC 15703, 94-BIM
B. longum BB536 (Morinaga Milk Industry)
B. longum SBT-2928 (Snow Brand Milk
Products)
B. breve Yakult
B. bifidus Bb-11
B. lactis (reclassied as B.
animalis)
Bb-12 (Chr. Hansen)
B. essensis Danone (Bioactivia)
B. lactis Bb-02
B. infantis Shirota
B. infantis Immunitass
B. infantis 744
B. infantis 01
B. laterosporus CRL 431
B. lactis LaftiTM, B94 (DSM)
B. longum UCC 35624 (UCCork)
B. lactis DR10/HOWARU DaniscoaAdapted from Krishnakumar and Gordon (2001); Holm (2003); Playne
et al. (2003); Shah (2004).more pH stable than L. acidophilus and producesbacteriocins against some enteropathogens. These proper-ties make this organism attractive as a probiotic. Frompublished reviews, four strains with the most publishedclinical data are L. rhamnosus GG, L. casei Shirota,B. animalis Bb-12, and Saccharomyces cerevisiae Boulardii(Shah, 2006b).
3. Health benets of functional probiotic cultures
A number of health benets are claimed in favourof products containing probiotic organisms includingantimicrobial activity and gastrointestinal infections, im-provement in lactose metabolism, antimutagenic proper-ties, anticarcinogenic properties, reduction in serumcholesterol, anti-diarrhoeal properties, immune systemstimulation, improvement in inammatory bowel diseaseand suppression of Helicobacter pylori infection (Kurmann& Rasic, 1991; Shah, 2000b, 2004). Some of the healthbenets are well established, while other benets haveshown promising results in animal models. However,additional studies are required in humans to substan-tiate these claims. Health benets imparted by probioticbacteria are strain specic, and not species- or genus-specic. It is important to note that no strain will pro-vide all proposed benets, not even strains of the samespecies, and not all strains of the same species will beeffective against dened health conditions. The strainsL. rhamnosus GG (Valio), S. cerevisiae Boulardii (Bioco-dex), L. casei Shirota (Yakult), and B. animalis Bb12(Chr. Hansen) have the strongest human health efcacydata with respect to management of lactose malabsorp-tion, rotaviral diarrhoea, antibiotic-associated diarrhoea,and Clostridium difficile diarrhoea (Playne, Bennet, &Smithers, 2003; Shah, 2006a, b). There is sufcient evi-dence to support the view that oral administration ofLactobacillus and Bifidobacterium is able to restore thenormal balance of microbial populations in the intestine(Shah, 2006b).
3.1. Antimicrobial activity and gastrointestinal infections
Probiotic bacteria produce organic acids, hydrogenperoxide and bacteriocins as antimicrobial substances thatsuppress the multiplication of pathogenic and putrefyingbacteria. Lactic and acetic acids account for over 90% ofthe organic acids produced. Lowering of pH due to lacticacid or acetic acid produced by these bacteria in the gut hasa bacteriocidal or bacteriostatic effect. Both Bifidobacter-ium and L. acidophilus show antagonistic effects towardsenteropathogenic Escherichia coli, Salmonella typhimurium,Staphylococcus aureus and Cl. perfringens. L. acidophilusproduces various bacteriocins and antibacterial substancessuch as Lactocidin, Acidolin, Acidophilin, Lactacium-Band inhibitory protein. Similarly, Bifidobacterium produces
ournal 17 (2007) 12621277Bidolin and Bilong, which inhibit several pathogenicbacteria (Shah, 1999).
-
ARTICLE IN PRESSiry J3.2. Effectiveness against diarrhoea
A major problem associated with antibiotic treatment isappearance of diarrhoea, often caused by Cl. difficile. Thisorganism is found in small numbers in the healthyintestine, but disruption of indigenous microora due toantibiotic treatment leads to an increase in their numberand toxin production, which causes the developmentof diarrhoea. Treatment with metonidazole or vancomycinis usually effective but recurrences are common. Probioticshave proved to be useful as a prophylactic regimenwith antibiotic-associated diarrhoea as well as for treat-ment after onset of antibiotic induced diarrhoea. A dailydose of Lactobacillus GG has been found to be effectivein termination of diarrhoea. Studies with a yeast prepara-tion containing S. cerevisiae Boulardii has also beeneffective in treatment of Cl. difficile related colitis (Shah,2004, 2006b).Rotavirus is one of the most common causes of acute
diarrhoea in children worldwide. During diarrhoeal stageof infection, the permeability of gut epithelial cells isincreased to intact proteins. Probiotics are claimed toshorten duration of rotavirus diarrhoea in children(Saavedra, Bauman, Oung, Perman, & Yolken, 1994).The strongest evidence of a benecial effect of denedstrains of probiotics has been established usingL. rhamnosus GG and B. lactis Bb-12 (now reclassied asB. animalis Bb-12) for prevention and treatment ofdiarrhoea and acute diarrhoea in children mainly causedby rotaviruses. Selected probiotic strains are also effectiveagainst antibiotic-associated diarrhoea. Certain probioticstrains can inhibit the growth and adhesion of a range ofenteropathogens. Studies have indicated benecial effectsagainst pathogens such as Sal. typhimurium and Sal.enteriditis. B. longum SBT-2828 has shown inhibition ofenterotoxigenic Escherichia coli. A pediatric beveragecontaining a mix of B. animalis, L. acidophilus, andL. reuteri has been found to be useful in the preventionof rotavirus diarrhoea (Guandalini et al., 2000).L. rhamnosus GG has been reported to be more effective
in treatment of rotavirus diarrhoea than preparationscontaining Str. thermophilus and L. delbrueckii ssp.bulgaricus. L. reuteri has also been effective in shorteningduration of rotavirus diarrhoea. It reduces the duration ofdiarrhoea in children suffering from rotavirus diarrhoea.Treatment with Lactobacillus GG was associated withenhancement of IgA-specic antibody-secreting cells torotavirus and serum IgA antibody level.There is also strong evidence that probiotic strains can
prevent travellers diarrhoea (Hilton, Kolakowski, Singer,& Smith, 1997), which is caused by bacteria, particularlyenterotoxigenic E. coli. Several studies have been carriedout to assess the effects of probiotic preparations asprophylaxis for travellers diarrhoea; however, the resultshave been conicting. In one study, Danish tourists on
N.P. Shah / International Daa 2-week trip to Egypt were given a mixture of livefreeze-dried preparation of L. acidophilus, B. animalis,L. delbrueckii ssp. bulgaricus and Str. thermophilus at adaily dose of 109 cfu. The administration of probioticpreparation reduced the frequency of diarrhoea. A similarstudy conducted with Finnish tourists using a lyophilizedpreparation of Lactobacillus GG also showed a reductionin the occurrence of travellers diarrhoea. On the otherhand, Katelaris, Salam, and Farthing (1995) and de diosPozo-Olano, Warram, Gomez, and Cacazos (1978) havefound no effect in people suffering from travellersdiarrhoea when L. fermentum KLD L. acidophilus andL. bulgaricus were given in separate studies.Yoghurt containing B. longum was found to be effective
in reducing the course of erythromycin induced diarrhoea.Faecal counts of Lactobacillus GG indicated that theorganisms colonized the intestine despite erythromycintreatment. Probiotic reparations containing 4 109 cfu ofB. animalis Bb-12 and L. acidophilus La-5 has shownsimilar results when volunteers received ampicillin alongwith probiotic preparation. Several studies have shown areduction in diarrhoea in subjects taking S. cerevisiaeBoulardii during the period of antibiotic treatment (Shah,2004, 2006a).
3.3. Improvement in lactose metabolism
Relief of the symptoms of lactose malabsorption isprobably the most widely accepted health benet ofprobiotic organisms. Lactose malabsorption is a conditionin which lactose, the principal carbohydrate of milk, is notcompletely hydrolysed into its component monosacchar-ides, glucose and galactose. Since lactose is cleaved into itsconstituent monosaccharides by the enzyme b-D-galactosi-dase, lactose malabsorption results from a deciency of thisenzyme. Lactose malabsorbers often complain of gastricdistress after consuming fresh, unfermented milk or milkproducts due to the formation of hydrogen gas bymicrobial action on undigested lactose in the gut (Shah,1993; Shah, Fedorak, & Jelen, 1992).The traditional cultures used in making yoghurt (i.e.,
L. delbrueckii ssp. bulgaricus and Str. thermophilus) containsubstantial quantities of b-D-galactosidase (Shah, 2000c),and so both yoghurt and probiotic yoghurt are toleratedwell by lactose malabsorbers. However, reduced levels oflactose in fermented products due to partial hydrolysis oflactose during fermentation are only partly responsible forthis greater tolerance for yoghurt. The slower gastricemptying of both semi-solid and pasteurized yoghurtresults in better absorption of lactose (in the latter, enzymeactivity and bacteria are destroyed due to heat treatment;Shah et al., 1992).
3.4. Antimutagenic properties
An antimutagenic effect of fermented milks has beendetected against a range of mutagens and promutagens in
ournal 17 (2007) 12621277 1265various test systems based on microbial and mammaliancells. Probiotic organisms are reported to bind mutagens
-
ARTICLE IN PRESSry Jto the cell surface (Orrhage, Sillerstrom, Gustafsson,Nord, & Rafter, 1994). Probiotic bacteria are reportedto reduce faecal enzymatic activities including b-glucur-onidase, azoreductase, and nitroreductase, which areinvolved in activation of mutagens (Goldin & Gorbach,1984b).Lankaputhra and Shah (1998) studied the antimutagenic
activity of organic acids produced by probiotic bacteriaagainst several mutagens and promutagens. The TA-100mutant of Sal. typhimurium (His) strain was used as amutagenicity indicator organism. The mutagenicity testwas carried out using the Ames Salmonella test. In theirstudy, butyric acid showed a broad-spectrum antimuta-genic activity against all mutagens or promutagens studiedand live bacterial cells showed higher antimutagenicitythan killed cells. Inhibition of mutagens and promutagensby probiotic bacteria was permanent for live cells andtemporary for killed cells. The results emphasized theimportance of consuming live probiotic bacteria and ofmaintaining their viability in the intestine to provideefcient inhibition of mutagens. Table 3 summarizesstudies pertaining to antimutagenic properties of functionalmicroorganisms.
3.5. Anticarcinogenic properties
Bacteria and metabolic products such as genotoxiccompounds (nitrosamine, heterocyclic amines, phenoliccompounds, and ammonia) are responsible for colorectalcancer. The consumption of cooked red meat, especiallybarbequed meat, and low consumption of bre arereported to play a major role in causing colorectal cancer.The colonic ora are also reported to cause carcinogenesismediated by microbial enzymes such as b-glucuronidase,azoreductase, and nitroreductase, which convert procarci-nogens into carcinogens.Certain strains of L. acidophilus and Bifidobacterium spp.
are reported to decrease the levels of enzymes such asb-glucuronidase, azoreductase, and nitroreductase respon-sible for activation of procarcinogens and consequen-tly decrease the risk of tumour development (Yoon,Benamouzig, Little, Francois-Collange, & Tome, 2000).Short chain fatty acids produced by L. acidophilus andBifidobacterium, L. plantarum and L. rhamnosus arereported to inhibit the generation of carcinogenic productsby reducing enzyme activities (Cenci, Rossi, Throtta, &Caldini, 2002).The anticarcinogenic effect of probiotic bacteria is
reported to be due to the result of removal of sources ofprocarcinogens (or the enzymes that lead to their forma-tion) improvement in the balance of intestinal microora,normalized intestinal permeability (leading to preventionor delaying of toxin absorption), strengthening of intestinalbarrier mechanisms, and activation of non-specic cellularfactors (such as macrophages and natural killer cells) via
N.P. Shah / International Dai1266regulation of g-interferon production. Orally administeredBifidobacterium is also reported to play a role in increasingproduction of IgA antibodies and functions of Peyerspatch cells (Singh et al., 1997). Table 4 summarizes studiespertaining to anticarcinogenic properties of functionalmicroorganisms.
3.6. Reduction in serum cholesterol
The level of serum cholesterol is a major factor forcoronary heart disease, and elevated levels of serumcholesterol, particularly LDL-cholesterol, have been linkedto an increased risk (Liong & Shah, 2006). There is a highcorrelation between dietary saturated fat or cholesterolintake and serum cholesterol level. Feeding of fermentedmilk containing very large numbers of probiotic bacteria(109 bacteria g1) to hypercholesterolaemic human sub-jects has resulted in lowering cholesterol from 3.0 to1.5 gL1.Probiotic bacteria are reported to de-conjugate bile salts:
deconjugated bile acid does not absorb lipid as readily asits conjugated counterpart, leading to a reduction incholesterol level. L. acidophilus is also reported to takeup cholesterol during growth and this makes it unavailablefor absorption into the blood stream (Klaver & Meer,1993). Studies showing reduction in serum cholesterol aresummarized in Table 5.
3.7. Helicobacter pylori infection
Helicobacter pylori is a pathogenic bacterium that causespeptic ulcers, type B gastritis and chronic gastritis and isnormally present in the stomach as an opportunisticpathogen without causing any symptoms (Armuzzi et al.,2001; Sakamoto et al., 2001).Antibiotic treatments can successfully eradicate
H. pylori. However, antibiotics often cause side effectsand make the bacteria more antibiotic resistant. Probioticorganisms do not appear to eradicate H. pylori, but theyare able to reduce the bacterial load in patients infectedwith H. pylori. L. johnsonii La1 and L. gasseri OLL2716have been found to reduce H. pylori colonization andinammation (Felley et al., 2001). Similarly, L. caseiShirota and L. acidophilus are reported to inhibit thegrowth of H. pylori (Cats et al., 2003). Studies showingprobiotic micro-organisms and H. pylori infection aresummarized in Table 6.
3.8. Inflammatory bowel disease
Inammatory bowel disease (ulcerative colitis andCrohns disease) is related to the intestinal microora.Symptoms of inammatory bowel disease include adisturbance in bowel habits and mucosal inammation.In the intestine of people with inammatory bowel diseasethe numbers of Lactobacillus and Bifidobacterium are lowerand those of coccoids and anaerobes are higher. Probiotics
ournal 17 (2007) 12621277do not cure the disease, but once patients are in remissionthrough treatment with corticosteroids, some probiotics
-
ARTICLE IN PRESSiry JTable 3
Antimutagenic properties of functional microorganisms
Microorganism Antimutagenic activity
N.P. Shah / International Dacan prolong the remission period, thus reducing theincidence of relapse and the use of corticosteroids. Thisimproves the quality of life of patients. Studies showingprobiotic micro-organisms and inammatory bowel diseaseare summarized in Table 7.
L. acidophilus strains Microbial binding leading
such as 4-nitroquinoline-N
L. acidophilus 2400, 2401, 2404, 2405, 2409,
2415; B. bifidum 1900, 1901, 1912, 1920, 1930
and 1941
Antimutagenic activity of
against mutagens and prom
and 2-amino-3-methyl-3H
L. casei CRL 431 The immune cells activated
mice were those of the inn
specic markers of these c
modication in the numb
L. rhamnosus GG Inuenced intestinal phys
modulation of the endoge
L. acidophilus (LAFTIs L10); B. lactis (LAFTI
B94)
Synbiotic combinations in
induced damage to coloni
L. acidophilus (SBT0274, SBT1703, SBT10239,
SBT10241); B. longum (SBT 2928)
Exhibited a high percentag
amino acid pyrolysates us
of Sal. typhimurium TA 9
L. bulgaricus 291; Str. thermophilus F4, V3; B.
longum BB536
Clear evidence for DNA-p
yoghurt production again
which are targets of tumo
nitrosoguanidine, 1,2-dim
azoxymethane and also re
pre-carcinogenic lesions in
Lactobacillus GG; L. delbrueckii subsp.
rhamnosus (LGG); B. lactis Bb12
The groups treated with p
the total number of tumou
(P 0.04) than in untreatprotective effect of probio
L. helveticus L89 Antimutagenic compound
by L. helveticus, and the r
contributing mechanism. H
variant of the same strain
nitroquinoline-N0-oxide (4
B. infantis CCRC 14633, B. longum B6,
L. acidophilus CCRC 14079 and Str.
thermophilus CCRC 14085
Unfermented soymilk exer
dimethyl-4-amino-bipheny
NQO), the fermented soy
against DMAB than 4-NQ
themophilus and B. infanti
antimutagenicity of 85.07%
and DMAB.
L. acidophilus LA 106 The milk cultured with L.
inhibition of 77% against
nitrosoguanidine among t
Probiotic bacterium Ent. faecium M-74 Exerted different antimuta
N0-nitro-N-nitrosoguanidiinduced mutagenicity in S
presence (+Se) or absence
antimutagenicity of MRS(
Se) extract. Selenium also
and killed cells of Ent. faeReferences
ournal 17 (2007) 12621277 12673.9. Immune system stimulation
The intestine is the bodys largest immune organ and theintestinal microora and the metabolic activity of theintestine is equivalent to that of the liver. Probiotics may
to inhibition of mutagens and promutagens0-oxide, 2-nitrouorene and benzopyrene.
Ayebo et al. (1982)
organic acids produced by probiotic bacteria
utagens such as 2-nitroourene, aatoxin-B,
-imidazoquinoline.
Lankaputhra and Shah
(1998)
after oral L. casei administration to BALB/c
ate immune response (with an increase in the
ells, CD-206 and TLR-2), with no
er of T cells.
Galdeano and Perdigon
(2006)
iology either directly or indirectly through
nous ecosystem or immune system.
Marteau et al. (2001)
regulating the consequences of carcinogen-
c epithelial cells in male Sprague-Dawley rats.
Le Leu et al. (2005)
e of antimutagenicity and binding in vitro on
ing a streptomycin-dependent (SD510) strain
8.
Sreekumar and Hosono
(1998)
rotective effects of lactobacilli used for
st DNA damage caused by HCAs in organs
r induction by 1-methyl-3-nitro-1-
ethylhydrazine, N-methyl-N-nitrosourea and
duce chemically induced-DNA migration and
colon cells of male Fischer 344 rats.
Zsivkovits et al. (2003);
Wollowski et al. (2001)
robiotics, the proportion of cancers relative to
rs in the same groups was signicantly lower
ed male F344 rats suggesting that the possible
tics is restricted to malignant tumours.
Femia et al. (2002)
s were produced in milk during fermentation
elease of peptides was one possible
owever, milk fermented by a non-proteolytic
showed no inhibitory effects on 4-
-NQO).
Matar et al. (1997)
ted lower antimutagenic activity against 3, 20-l (DMAB) than 4-nitroquinoline-N-oxide (4-
milk, showed a higher antimutagenic activity
O. Soymilk fermented with both Str.
s simultaneously exhibited the highest
and 85.78%, respectively, against 4-NQO
Hsieh and Chou (2006)
acidophilus LA 106 (LA2) showed the highest
the mutagenicity of N-methyl-N-nitro-N-
he 71 strains tested.
Hosoda et al. (1992)
genic activity against ooxacin-, N-methyl,
ne- and sodium 5-nitro-2-furylacrylate-
al. typhimurium assay depending on the
of disodium selenite pentahydrate (-Se). The
+Se) extract was higher than that of MRS(-
enhanced the antimutagenic effect of both live
cium M-74, respectively.
Belicova et al. (2004)
-
ARTICLE IN PRESS
ion
t-b
xpe
the
FM
ts h
, da
ld r
uc
bio
oxi
tab
n c
e a
ucta
ry JTable 4
Probiotic microorganisms and their anticarcinogenic properties
Microorganisms Anticarcinogenic funct
L. acidophilus NCFM Rats consuming a mea
cancer after a 20-wk e
supplement increased
Administration of NC
tumours. NCFM fed ra
the faeces.
L. acidophilus NCFM Using human subjects
resulted in a 2- to 4-fo
glucuronidase, nitrored
L. bulgaricus; Str. thermophilus Ingestion of viable pro
detoxication of genot
L. bulgaricus; Str. thermophilus Possibly inuenced me
the colon.
Lactobacillus and Bifidobacterium; cellular
components and metabolites of LAB
Limits DNA damage i
Bifidobacterium fermented milk; fermented milk
with L. acidophilus, B. bifidum, Lc. lactis ssp.
lactis, Lc. lactis ssp. cremoris
Procarcinogenic enzym
nitroreductase, azored
N.P. Shah / International Dai1268directly or, by changing the composition or activity of theintestinal microora, indirectly inuence the bodys im-mune function (Marteau et al., 1997). Probiotic culturesproduce g-interferon by T-cells and stimulate cytokines asrepresented by TNF-a (tumour necrosis factor) and IL-6and IL-10 (interleukines 6 or 10). Immunomodulation byL. acidophilus and Bifidobacterium has been observed, inparticular IgA levels and non-specic immunity. Ingestionof probiotic yoghurt has been reported to stimulatecytokine production in blood cells and enhance theactivities of macrophages. Studies showing probioticmicro-organisms and immune system stimulation aresummarized in Table 8.
4. Prebiotics and synbiotics
To maximize effectiveness of bidus products, prebioticsare used in probiotic foods. A prabiotic is a non-digestiblefood that benecially affects the host by selectivelystimulating the growth and/or activity of one or a limitednumber of bacteria in the colon (Shah, 2004, 2006b).Some oligosaccharides, due to their chemical structure,
are resistant to digestive enzymes and therefore pass intothe large intestine where they become available for
L. acidophilus; Lc. lactis ssp. cremoris, Binding of mutagens.
B. lactis; L. gasseri Released bioactive hydrox
showed anticarcinogenic p
L. acidophilus Decreased polyps, adenom
L. acidophilus 145; B. longum 913 Reduction of the risk of c
DNA damage in animals.References
ased diet showed a lower incidence of colon
rimental period, implying that the NCFM
latency period for colon cancer in rats.
along with antibiotics also decreased colon
ad a signicantly lower level of free amines in
Goldin and Gorbach
(1980, 1984a, c).
ily consumption of milk containing NCFM
eduction in the activity of faecal enzymes, X -tase, and azoreductase.
Goldin and Gorbach
(1984b, c)
tics led to anticarcinogenic effects, through
ns in the gut of rats.
Wollowski et al.
(2001a, b)
olic, immunologic, and protective functions in Kasper (1996)
olon cells (antigenotoxicity). Pool-Zobel et al. (1993,
1996); Wollowski
(1998); Ji (1997)
ctivity decreased: b-glucuronidase,se and detoxifying enzyme activity increased.
Benno and Mitsuoka
(1992); Bouhnik et al.
(1996)
ournal 17 (2007) 12621277fermentation by saccharolytic bacteria. Compounds thatare either partially degraded or not degraded by the hostand are preferentially utilized by Bifidobacterium as acarbon and energy sources are referred to as bidogenicfactors. Some bidogenic factors that are of commercialsignicance include fructo-oligosaccharides, lactose deri-vatives such as lactulose, lactitol, galacto-oligosaccharides,and soyabean oligosaccharides. Resistant starch and non-starch oligosaccharides are classied as colonic foods, butnot as prebiotics, because they are not metabolized bycertain benecial bacteria (Shah, 2004).The products that contain both prebiotics and probiotics
are referred to as synbiotics. Synbiotics are a combinationof the effects of probiotics and prebiotics to produce healthenhancing functional food ingredients. Japan is the worldleader in probiotic and prebiotic products. The majority ofyoghurts marketed in Australia, USA and Europe in recentyears contain one or more species of probiotic bacteria andsome form of prebiotics (Shah, 2000a).
5. Alternate products for incorporating probiotics
To realize health benets, probiotic bacteria must beviable and must be available in a high concentration,
Orrhage et al. (1994);
Morotomi and Mutai
(1986)
ycinnamic acids in the human colon which
roperties both in vitro and in animal models.
Couteau, et al. (2001)
as and colon cancer in experimental animals. Gorbach et al. (1987)
olon cancer by inhibiting carcinogen-induced Moschner et al. (2004)
-
ARTICLE IN PRESS
l le
l as
of
men
iry JTable 5
Probiotic microorganisms and reduction in serum cholesterol
Microorganisms Probiotic function
L. acidophilus Lowered serum cholestero
L. acidophilus; L. casei ASCC 1520, ASCC
1521, ASCC 292, ATCC 15820, and
L. acidophilus ATCC 4962
Highest in vitro cholestero
cholesterol in the presence
media.
Human strain of Ent. faecium 29 men receiving milk fer
N.P. Shah / International Datypically 106 cfu g1 of a product. Despite the importanceof viability, studies have shown low populations ofprobiotic bacteria in probiotic foods (Anon, 1992; Shah,Lankaputhra, Britz, & Kyle, 1995; Shah, Ali, & Ravula,2000). It is questionable whether such products can providethe claimed benets if the populations of probiotic bacteriaare low. Yoghurt is considered the most important carrierof probiotic bacteria. However, a number of factors affectthe loss of viability of probiotic organisms in yoghurt,including acidity of products, acid produced duringrefrigerated storage (also known as post-acidication),
(1081011 cfuL1) led to a dec0.370.41mmolL1 after 6wkno effect.
Enterococcus group of 87 men
decrease in serum LDL conce
than the placebo-group at we
Ent. faecium (Gaio) Was found to be effective in r
lipoprotein cholesterol by 4%
control group.
L. casei TMC 0409 In a single blind parallel stud
(200mLday1) showed signicholesterol, compared with th
supplementation. The levels of
in subjects receiving the ferme
Str. thermophilus TMC 1543 In a single blind parallel stud
(200mLday1) showed signicholesterol, compared with th
supplementation. The levels of
in subjects receiving the ferme
B. longum 913; L. acidophilus 145 Consumption of 300 g yoghur
seven weeks increased high de
0.3mmolL1 (p 0.002) andlipoprotein from 3.24 to 2.48
L. gasseri SBT0270, SBT0274 Exerted hypocholesterolaemic
through deconjugation of bile
L. reuteri CRL 1098 (104 cells d1) Caused a 40% reduction in trhigh density lipoprotein to lo
translocation of the native mi
Albino mice.
B. longum BL1 Led to a signicant lowering
cholesterol, low-density lipop
products compared with the c
lipoprotein cholesterol concen
L. brevis NR1C1684; Ent. faecalis Assimilated more cholesterol (
in the media with 0.2 and 0.4References
vels. Ouwehand et al. (2002)
similation of more than 25mgmL1
cholic acid, compared to other different bile
Liong and Shah (2005)
ted with a human strain of E. faecium Agerbaek, Gerdes, and
ournal 17 (2007) 12621277 1269level of oxygen in products and oxygen permeationthrough the package, sensitivity to antimicrobial sub-stances produced by starter bacteria and lack of nutrientsin milk (Dave & Shah, 1997a, b; Lankaputhra & Shah,1995, 1997; Shah, 2000a; Tamime, Saarela, Sandergaard,Mistry, & Shah, 2005).A number of food products including probiotic yoghurt,
ice cream, frozen fermented dairy deserts, and freeze-driedyoghurt (Capela, Hay, & Shah, 2006) have been employedas alternate delivery vehicles for probiotics. But, due to thepresence of high oxygen content (Lankaputhra & Shah,
rease in serum cholesterol by
, whereas consumption of the placebo had
Richelsen (1995)
and women had a signicantly larger
ntrations decreased throughout the study,
eks 4 and 12 (Po0.05).
Richelsen et al. (1996)
educing both total and low density
and 5%, respectively, compared with the
Agerholm-Larsen, Bell,
Grunwald, and Astrup
(2000)
y, subjects consuming fermented milk
cant increases of high density lipoprotein
e pre-intervention levels after four weeks
triglycerides were also reduced signicantly
nted milk.
Kawase et al. (1999)
y, subjects consuming fermented milk
cant increases of high density lipoprotein
e pre-intervention levels after eight weeks
triglycerides were also reduced signicantly
nted milk.
Kawase et al. (1999)
t supplemented with the 2 microrganisms for
nsity lipoprotein concentration by
decrease ratio of low to high density
(p 0.001).
Kiebling et al. (2002)
effect in rats fed a diet high in cholesterol
salts.
Usman and Hosono
(2000)
iglycerides and a 20 increase in the ratio of
w density lipoprotein without bacterial
croora into the spleen and liver of Swiss
Taranto et al. (1998)
of the serum concentrations of total
rotein cholesterol, and triglycerides in milk
ontrol, while no change in high-density
tration was observed.
Xiao et al. (2003)
in vivo) than the average of the other strains
% (wt/vol) oxgall, respectively.
Pereira and Gibson
(2002)
-
ARTICLE IN PRESS
H.
n 3
am
hibi
10
he i
ntib
ss t
han
tra
ry JTable 6
Probiotic microorganisms and Helicobacter pylori infection
Microorganisms Probiotic function
L. salivarius Suppressed and eradicated
by lactic acid secretion.
L. gasseri OLL2716(LG21) Signicant improvement o
reduced gastric mucosal in
Bac. subtilis 3 Inhibited H. pylori, the in
pylori isolates ranged from
Bac. clausii Bacteriotherapy reduced t
related to anti-H. pylori a
L. rhamnosus GG, L. rhamnosus LC705;
B. breve Bb99; Propionibacterium
freudenreichii ssp. shermanii JS
Probiotic group showed le
the total symptom score c
eradication therapy in con
N.P. Shah / International Dai12701997) in ice cream type products and injury due to freezingand freeze drying, many of the above mentioned productshave failed to successfully deliver the required level ofviable cells of probiotics. Cheese-based dips could be adelivery vehicle for probiotic bacteria because of its stablepH, the buffering capacity of ingredients used and thepresence of prebiotics (Tharmaraj & Shah, 2004).Other foods such as Cheddar cheese have also been
studied as carriers of probiotic microorganisms. Cheeseshave a number of advantages over fresh fermentedproducts such as yoghurt as a delivery system for viableprobiotic to gastrointestinal tract as they tend to havehigher pH, more solid consistency and relatively higher fatcontent. These offer protection to probiotic bacteria duringstorage and passage through the gastrointestinal tract.Cheeses also have higher buffering capacity than yoghurt.Cheddar cheeses, however, have long ripening time hence
Ent. faecium; Bac. subtilis; Bifidobacterium Produced heat-stable proteina
growth of both antibiotic-resi
L. acidophilus LA5+B. lactis Bb12 Increased eradication ofH. py
with combined use of certain
L. reuteri ATCC 55730 (108CFU) Probiotic supplemented childr
score during eradication thera
L. acidophilus strain LB (lyophilized) 1-week standard therapy incr
group of 120 patients.
L. casei strain Shirota There was a trend towards su
patients.
L. johnsonii La1 There was reduced density ofH
activity among 52 patients.
L. acidophilus NAS Eradication of H. pylori in six
L. casei subsp. rhamnosus (GG); S.
cerevisiae Boulardii; L. acidophilus; B. lactis
Probiotic preparations supple
were associated with lower inc
better treatment tolerability c
L. reuteri strains (JCM1112, JCM1081,
JCM1084 JCM2762, JCM2763 and
JCM2764)
Inhibition by selected L. reute
early stage of colonization inH
strains sharing glycolipid spec
probiotics.References
pylori in tissue cultures and animal models Aiba et al. (1998)
1 subjects infected with H. pylori and
mation following LG21 treatment.
Sakamoto et al. (2001)
tory zone for each of the 21 strains of H.
to 16mm.
Pinchuk et al. (2001)
ncidence of the most common side-effects
iotic therapy compared with placebo.
Nista et al. (2004)
reatment-related symptoms as measured by
ge (P 0.038) throughout the H. pylorist to the placebo group.
Myllyluoma et al. (2005)
ournal 17 (2007) 12621277development of probiotic Cheddar cheese requires a carefulexamination of the suitability of particular strain(s) tomaintain viability throughout the ripening and shelf life(Ong, Henriksson, & Shah, 2006). Two Cheddar cheesescontaining the probiotic DR20
TM
were launched by Main-land (Fonterra) in Australia in 2002 under the name InnerBalance.Another product, which has received much attention, is
soy based isoavone phytoestrogens, found abundantly insoybeans, are an isomeric family of di-phenolic compoundswith structural and functional similarities to humanestrogens. Soybeans and non-fermented soy foods (includ-ing soy extract) contain 8095% of their isoavones asglucoside conjugates, which are biologically inactive andnon-bioavailable (Tsangalis, Ashton, McGill, & Shah,2002). The biologically active (estrogen-like) and bioavail-able aglycone forms of daidzein, genistein and glycitein are
ceous compounds capable of inhibiting the
stant and -sensitive strains of H. pylori.
Tsai et al. (2004); Pinchuk
et al. (2001); Collado et
al. (2005)
lori among 160 dyspeptic subjects in 4 weeks
antibiotics.
Sheu et al. (2002)
en had a signicant reduction of GSRS
py compared to those receiving placebo.
Lionetti et al. (2006)
eased eradication rate of H. pylori in active Canducci et al. (2000)
ppressive effect in an active group of 20 Cats et al. (2003)
. pylori, reduced inammation and gastritis Felley et al. (2001)
of 14 patients after 8 weeks of treatment. Mrda et al. (1998)
menting a standard antiH. pylori regimen
idence of self-reported side effects and with
ompared to placebo.
Cremonini et al. (2002)
ri strains helped to prevent infection in an
. pylori leading to a proposal that L. reuteri
icity with H. pylori have a potential as
Mukai et al. (2002)
-
ARTICLE IN PRESS
a fr
atie
ma
teria
itab
l sy
roen
arrh
oxy
ciat
fun
e.
epis
he 2
e we
diar
se.
iry JTable 7
Probiotic microorganisms and inammatory bowel disease
Microorganisms Probiotic function
Ent. faecium Decreased duration of acute diarrhoe
L. acidophilus Signicant decrease of diarrhoea in p
L. plantarum Especially effective in reducing inam
enterocolitis in rats, small bowel bac
Reduced pain and constipation of irr
atulence, and pain in irritable bowe
L. reuteri Shortened the duration of acute gast
Shortened acute diarrhoea.
S. cerevisiae Boulardii (yeast) Reduced recurrence of Cl. difficile di
Effects on Cl. difficile and Klebsiella
shortened duration of antibiotic-asso
acute gastroenteritis. Decreased only
symptoms of irritable bowel syndrom
B. bifidum; Str. thermophilus Reduction of rotavirus shedding and
Ent. faecium PR88 Symptomatic improvement in 19 of t
caused by food intolerance for twelv
weight.
S. cerevisiae Boulardii; L.
rhamnosus GG
Helped prevent antibiotic-associated
appeared useful for Cl. difficile disea
N.P. Shah / International Dalinked to the prevention and potential treatment ofhormone-dependent disorders, including osteoporosis,cardiovascular disease and cancer (Tsangalis, Ashton,Wilcox, Shah, & Stojanovska, 2005). Recent research hasshown that isoavone aglycones are absorbed faster and inhigher amounts in humans than their respective glucosides.Intestinal microora, consisting predominantly of Bifido-bacterium, are of signicant importance in hydrolysingingested isoavone glucosides into bioavailable, bioactiveaglycones (Tsangalis et al., 2005), as well as in convertingdaidzein to the biologically potent equol. Fermentation ofplain soy extract with Bifidobacterium sp. is reported tohydrolyse the isoavone glucosides into bioactive agly-cones, increasing the concentration of aglycones from lessthan 10% to approximately 50% of the total isoavonecontent (Tsangalis et al., 2002).
5.1. Products based on bioactive peptides released from milk
proteins by proteolytic probiotics
The functionality of dairy proteins is further enhancedupon liberation of bioactive peptides by proteolysis(Gobbetti, Stepaniek, De Angelis, Corsetti, & Di Cagno,2002). Proteolysis is performed by naturally occurringenzymes in milk, starter cultures and the enzymes of thedigestive tract, resulting in release of many peptides withdifferent biogenic activities such as opioid, hypotensive,immunomodulating, antithrombotic and antimicrobialactivities.References
om gastroenteritis. Marteau et al. (2001)
nts receiving pelvic irradiation. Marteau et al. (2001)
tion in inammatory bowel; e.g.,
l overgrowth in children, pouchitis.
Schultz and Sartor (2000);
Vanderhoof (2000)
le bowel syndrome. Reduced bloating,
ndrome in controlled trial.
Nobaek et al. (2000)
teritis. Marteau et al. (2001)
Shornikova et al. (1997a,
1997b)
oea. Pochapin (2000)
toca resulted in decreased risk and/or
ed diarrhoea. Shortened the duration of
ctional diarrhoea, but not any other
Marteau et al. (2001)
odes of diarrhoea in children in hospital. Saavedra et al. (1994)
8 patients with high volume diarrhoea
eks and a signicant decrease in faecal
Hunter et al. (1996)
rhoea (AAD). S. cerevisiae Boulardii McFarland (2006)
ournal 17 (2007) 12621277 1271Dairy cultures used in production of fermented milkproducts have appreciable proteolytic activity enablingtheir rapid growth in milk. During fermentation, milkproteins, namely caseins, undergo a slight proteolyticdegradation resulting in a number of potentially bioactivepeptides (Table 9). Strains of L. helveticus have beenidentied as highly proteolytic and peptidolytic towardscaseins and their derivatives and, if used in fermentations,they can produce products with a range of bioactivitiessuch as opioid and hypotensive activity (Matar, Amiot,Savoie, & Goulet, 1996).Another group of bioactive peptides, termed angiotensin
I-converting enzyme (ACE) inhibitors, have been exten-sively studied due to their hypotensive role (Conlin et al.,2000). Milk proteins contain a number of ACE inhibit-ing peptides encrypted within their primary structures(Donkor, Henriksson, Vasiljevic, & Shah, 2005). Thesepeptides are liberated by proteolytic action of extracellularand intracellular enzymes of LAB proteolytic system.Strains of L. helveticus appeared to be superior in regardto production of ACE inhibitory peptides in comparisonwith other species tested (Ashar & Chand, 2003). Severalcommercial products containing highly proteolytic strainsof L. helveticus have been developed and marketed aspossessing hypotensive activity, including Calpiss andEvoluss. Calpiss sour milk (Calpis Co. Ltd., Tokyo,Japan), is prepared by fermenting milk using mixed culturecontaining L. helveticus CM4 (CP790) and S. cerevisiae.These organisms are responsible for the release of a range
-
ARTICLE IN PRESS
Table 8
Probiotic microorganisms and immune system stimulation
Microorganisms Probiotic function References
Lactobacillus strains Microbial interference therapythe use of nonpathogenic bacteria to
eliminate pathogens and as an adjunct to antibiotics.
Bengmark (2000)
Improved mucosal immune function, mucin secretion and prevention of
disease.
MacFarlane and
Cummings (2002)
L. acidophilus NCFM Prevented urogenital infection with subsequent exposure to three pathogens
E. coli, Klebsiella pneumoniae, Pseudomonas aeruginosa.
Sanders and Klaenhammer
(2001)
L. plantarum Positive effect on immunity in HIV+ children. Walker (2000)
L. rhamnosus Enhanced cellular immunity in healthy adults in controlled trial. Tomioka et al. (1992)
S. cerevisiae Boulardii In HIV associated acute diarrhoea, 56% of patients who were treated with
S.cerevisiae Boulardii had their symptoms resolved compared with only 6% of
placebo treated patients.
Saint Marc et al. (1999)
L. casei sp.; L. fermentum sp. Can induce cytokine responses in human peripheral blood mononuclear cells. Chen et al. (1999);
Miettinen et al. (1996)
L. salivarius strain Produces a large amount of lactic acid and completely inhibits the growth of
H. pylori in a mixed culture, hence suppressing H. pylori, and reducing the H.
pylori-induced inammatory response.
Aiba et al. (1998)
L. acidophilus DDS-1 Has been shown to induce the production of IL-1a and TNF- a. Rangavajhyala et al. (1997)
L. helveticus Able to release peptide compounds that may have important implications for
the modulation of the cellular immune response.
Matar et al. (2001);
Leblanc et al. (2002)
L. acidophilus TMC 0356 Signicantly increased the production of IL-6, IL-10, IL-12, and TNF-a Morita et al. (2002)
L. rhamnosus sp.; L. plantarum or
L. paracasei ssp. paracasei strains
Induced high levels of IL-10 in human blood mononuclear cells, or monocytes
and highest levels of IL-12 produced by L. paracasei subsp. paracasei strains,
implying its suitability to stimulate cell-mediated immunity.
Hessle et al. (1999)
L. casei Shirota Induced a marked increase in the production of IL-12, probably by
macrophages, which in turn stimulated the production of IFN-g.Kato et al. (1999)
Induced the production of several cytokines, such as IFN- g, IL-1b and TNF-a in mice.
Matsuzaki (1998)
B. adolescentis; B. longum Induced signicantly more pro-inammatory cytokine secretion of IL-12 and
TNF-a than did the infant-type bidobacteria, B. bifidum, B. breve, and B.infantis.
He et al. (2002)
L. plantarum 299v In the presence of the proinammatory cytokine TNF-a, exerted a protectiveeffect by downregulating IL-8 secretion in the human HT-29 colonic epithelial
cell line.
Mccracken et al. (2002)
L. casei DN-114 001, DN-114056,
ATCC-334; L. bulgaricus LB-10 strains
Normal colonic specimens have been obtained from Crohns disease (CD)
patients with neoplasm and inamed ileal specimens after being cultured with
either of these microorganisms.
Borruel et al. (2003)
L. johnsonii La1 Increases transforming growth factor beta mRNA in leukocyte-sensitized
Caco-2 cells.
Haller et al. (2000)
L. johnsonii La1; B. bifidum Bb12
strains
In healthy volunteers receiving a fermented milk product supplemented with
either bacteria for 3 weeks, phagocytosis of E. coli sp. by leukocytes isolated
from the blood was enhanced.
Schiffrin et al. (1995, 1997)
L. casei Shirota; B. breve YIT4064
strains
In mice, oral administration of either bacteria activated the humoral immune
system.
Yasui et al. (1999)
L. casei ssp. rhamnosus GG Prevents cytokine-induced apoptosis in intestinal epithelial cell lines in culture
by activating the anti-apoptotic Akt/protein kinase B, and by inhibiting the
pro-apoptotic p38/mitogen-activated protein kinase by stimulating the
production of TNF-a, IL-1b, IL-1a, or INF-g.
Yan and Polk (2002)
N.P. Shah / International Dairy Journal 17 (2007) 126212771272
-
lar. Other probiotic organisms including Ent. faecium,
ARTICLE IN PRESS
thei
Leu-Asn-Val-Pro-Gly- L. delbrueckii ssp.
iry JS. cerevisiae Boulardii and Propionibacterium have poten-tial to be used in probiotic products. Several health benetshave been claimed for probiotic bacteria. Yoghurt is themost important delivery vehicle for probiotic organisms.of potent bioactive peptides including two tripeptides, Val-Pro-Pro and Ile-Pro-Pro (Nakamura et al., 1995; Vasiljevic& Shah, 2007).
6. Conclusions
Probiotic products containing L. acidophilus, Bifidobac-terium spp. and L. casei are becoming increasingly popu-
glu-Ile-Val-Glu bulgaricus SS1
Asn-Ile-Pro-Pro-Leu- Lc. lactis ssp.
Thr-Glu-Thr-Pro-Val cremoris FT4
aAdapted from Vasiljevic and Shah (2007).Table 9
Some examples of the identied bioactive peptides in fermented milk and
Sequence Microbial agent
Val-Pro-Pro L. helveticus CM4 &
Ile-Pro-Pro S. cerevisiae
Val-Pro-Pro L. helveticus
Ile-Pro-Pro LBK16H
Asn-Leu-His-Leu-Pro- L. helveticus NCC
Leu-Pro-Leu-Leu 2765
Tyr-Pro-Phe-Pro-Glu- L. helveticus NCC
Pro-Ile-Pro-Asn 2765
Tyr-Pro L. helveticus CPN4
N.P. Shah / International DaCheddar cheese, dips and spreads are becoming popular asalternate products for incorporation of probiotics. Proteo-lytic strains of probiotic bacteria are used to releasebioactive peptides such as ACE-inhibitor peptides forfurther improving the health benets of probiotic foods.
References
Agerbaek, M., Gerdes, L. U., & Richelsen, B. (1995). Hypocholester-
olaemic effect of a new fermented milk product in healthy middle-aged
men. European Journal of Clinical Nutrition, 49, 346352.
Agerholm-Larsen, L., Bell, M. L., Grunwald, G. K., & Astrup, A. (2000).
The effect of probiotic milk product on plasma cholesterol: A meta-
analysis of short-term intervention studies. European Journal of
Clinical Nutrition, 54, 856860.
Aiba, Y., Suzuki, N., Kabir, A. M., Takagi, A., & Koga, Y. (1998). Lactic
acid-mediated suppression of Helicobacter pylori by the oral admin-
istration of Lactobacillus salivarius as a probiotic in a gnotobiotic
murine model. American Journal of Gastroenterology, 93, 20972101.
Anon. (1992). Yoghurt and probiotics. Choice, 11, 3235.
Armuzzi, A., Cremonini, F., Bartolozzi, F., Canducci, F., Candelli, M.,
Ojetti, V., et al. (2001). The effect of oral administration of
Lactobacillus GG on antibiotic-associated gastrointestinal side-effects
during Helicobacter pylori eradication therapy. Alimentary Pharmacol-
ogy and Therapeutics, 15, 163169.Ashar, M. N., & Chand, R. (2003). ACE-inhibitory activity of lactic acid
bacteria in fermented milks. MilchwissenschaftMilk Science Interna-
tional, 58, 5961.
Ayebo, A. D., Shanani, K. M., Dam, R., & Friend, B. A. (1982). Ion
exchange separation of the antitumor components in yogurt dialyzate.
Journal of Dairy Science, 65, 23882390.
Belicova, A., Krizkova, L., Dobias, J., Krajcovic, J., & Ebringer, L.
(2004). Synergistic activity of selenium and probiotic bacterium
Enterococcus faecium M-74 against selected mutagens in Salmonella
assay. Folia Microbiologica, 49, 301305.
Bengmark, S. (2000). Colonic food: Pre- and probiotics. American Journal
of Gastroenterology, 95, S5S7.
Benno, Y., & Mitsuoka, T. (1992). Impact of Bifidobacterium longum on
human fecal microora. Microbiology and Immunology, 36, 683694.
Borruel, N., Casellas, F., Antolin, M., Llopis, M., Carol, M., Espiin, E.,
r physiological activitya
Precursor Bioactivity
b- & k-casein Hypotensive
b- & k-casein Hypotensive
b-casein ACE inhibition
b-casein Opioid
caseins ACE inhibition
b-casein ACE inhibition
b-casein ACE inhibition
ournal 17 (2007) 12621277 1273et al. (2003). Effects of nonpathogenic bacteria on cytokine secretion
by activity of the probiotic strain Bacillus subtilis 3 is due to secretion
of antibiotics. Antimicrobial Agents and Chemotherapy, 45, 31563161.
Bouhnik, Y., Flourie, B., Andrieux, C., Bisetti, N., Briet, F., & Rambaud,
J.-C. (1996). Effects of Bifidobacterium sp fermented milk ingested with
or without inulin on colonic bidobacteria and enzymatic activities in
healthy humans. European Journal of Clinical Nutrition, 50, 269273.
Canducci, F., Armuzzi, A., Cremonini, F., Cammarota, G., Bartolozzi, F.,
Pola, P., et al. (2000). A lyophilized and inactivated culture of
Lactobacillus acidophilus increases Helicobacter pylori eradication
rates. Alimentary Pharmacology and Therapeutics, 14, 16251629.
Capela, P., Hay, T. K. C., & Shah, N. P. (2006). Effect of cryoprotectants,
prebiotics and microencapsulation on survival of probiotic organisms
in yoghurt and freeze-dried yoghurt. Food Research International, 39,
203211.
Cats, A., Kulpers, E. J., Bosschaert, M. A., Pot, R. G., Vandenbroucke-
Gauls, C. M., & Kusters, J. G. (2003). Effect of frequent consumption
of Lactobacillus casei-containing milk drink in Helicobacter pylori-
colonised subjects. Pharmacology and Therapeutics, 17, 429435.
Cenci, G., Rossi, J., Throtta, F., & Caldini, G. (2002). Lactic acid bacteria
isolated from dairy products inhibit genotoxic effect of 4-nitroquino-
line-1-oxide in SOS-chromotest. Systematic and Applied Microbiology,
25, 483490.
Chen, T., Isomaki, P., Rimpilainen, M., & Toivanen, P. (1999). Human
cytokine responses induced by gram-positive cell walls of normal
intestinal microbiota. Clinical and Experimental Immunology, 118,
261267.
-
ARTICLE IN PRESSry JCollado, M. C., Gonzalez, A., Gonzalez, R., Hernandez, M., Ferrus, M.
A., & Sanz, Y. (2005). Antimicrobial peptides are among the
antagonistic metabolites produced by Bifidobacterium against Helico-
bacter pylori. International Journal of Antimicrobial Agents, 25,
385391.
Conlin, P. R., Chow, D., Miller, E. R., Svetkey, L. P., Lin, P. H., Harsha,
D. W., et al. (2000). The effect of dietary patterns on blood pressure
control in hypertensive patients: Results from the dietary approaches
to stop hypertension (DASH) trial. American Journal of Hypertension,
13, 949955.
Couteau, D., McCartney, A. L., Gibson, G. R., Williamson, G., & Faulds,
C. B. (2001). Isolation and characterization of human colonic bacteria
able to hydrolyse chlorogenic acid. Journal of Applied Microbiology,
90, 873881.
Cremonini, F., Di Caro, S., Covino, M., Armuzzi, A., Gabrielli, M.,
Santarelli, L., et al. (2002). Effect of different probiotic preparations
on anti-Helicobacter pylori therapy-related side effects: A parallel
group, triple blind, placebo-controlled study. American Journal of
Gastroenterology, 97, 27442749.
Curry, B., & Crow, V. (2003). Lactobacillus spp.: General characteristics.
Encyclopedia of Dairy Sciences, 3, 14791484.
Dave, R. I., & Shah, N. P. (1997a). Effect of level of starter culture on
viability of yoghurt and probiotic bacteria in yoghurts. Food Australia,
49, 164168.
Dave, R. I., & Shah, N. P. (1997b). Viability of yoghurt and probiotic
bacteria in yoghurts made from commercial starter cultures. Interna-
tional Dairy Journal, 7, 3141.
de dios Pozo-Olano, J., Warram, J. H., Gomez, R. G., & Cavazos, M. G.
(1978). Effect of lactobacilli preparation on travellers diarrhoea. A
randomized, double blind clinical trial. Gastroenterology, 74, 829830.
Donkor, O. N., Henriksson, A., Vasiljevic, T., & Shah, N. P. (2005).
Probiotic strains as starter cultures improve angiotensin-converting
enzyme inhibitory activity in soy yogurt. Journal of Food Science, 70,
375381.
Felley, C. P., Corthesy-Theulaz, I., Rivero, J. L., Sipponen, P., Kaufmann,
M., Bauerfeind, P., et al. (2001). Favourable effect of an acidied milk
(LC-1) on Helicobacter pylori gastritis in man. European Journal of
Gastroenterology and Hepatology, 13, 2529.
Femia, A. P., Luceri, C., Dolara, P., Giannini1, A., Biggeri, A., Salvadori,
M., et al. (2002). Antitumorigenic activity of the prebiotic inulin
enriched with oligofructose in combination with the probiotics
Lactobacillus rhamnosus and Bifidobacterium lactis on azoxymethane-
induced colon carcinogenesis in rats. Carcinogenesis, 23, 19531960.
Fuller, R. (1992). History and development of probiotics. In R. Fuller
(Ed.), Probiotics, the scientific basis (pp. 18). London, UK: Chapman
& Hall.
Galdeano, C. M., & Perdigon, G. (2006). The probiotic bacterium
Lactobacillus casei induces activation of the gut mucosal immune
system through innate immunity. Clinical Vaccine and Immunolology,
13(2), 219226.
Gobbetti, M., Stepaniak, L., De Angelis, M., Corsetti, A., & Di Cagno, R.
(2002). Latent bioactive peptides in milk proteins: Proteolytic
activation and signicance in dairy processing. Critical Reviews in
Food Science and Nutrition, 42, 223239.
Goldin, B. R., & Gorbach, S. L. (1980). Effect of Lactobacillus acidophilus
dietary supplements on 1,2-dimethylhydrazine dihydrochloride-in-
duced intestinal cancer in rats. Journal of the National Cancer Institute,
64, 63265.
Goldin, B. R., & Gorbach, S. L. (1984a). Alterations of the intestinal
microora by diet, oral antibiotics, and Lactobacillus: Decreased
production of free amines from aromatic nitro compounds, azo dyes,
and glucuronides. Journal of the National Cancer Institute, 73,
689695.
Goldin, B. R., & Gorbach, S. L. (1984b). The effect of milk and
Lactobacillus feeding on human intestinal bacterial enzyme activity.
American Journal of Clinical Nutrition, 39, 756761.
N.P. Shah / International Dai1274Goldin, B. R., & Gorbach, S. L. (1984c). The effect of oral administration
of Lactobacillus and antibiotics on intestinal bacterial activity andchemical induction of large bowel tumors. Developmental Industrial
Microbiology, 25, 139150.
Gorbach, S. L., Chang, T. W., & Goldin, B. (1987). Successful treatment
of relapsing Clostridium difficile colitis with Lactobacillus GG. Lancet,
2, 1519 (letter).
Guandalini, S., Pensabene, L., Zikri, M. A., Dias, J. A., Casali, L. G.,
Hoekstra, H., et al. (2000). Lactobacillus GG administered in oral
rehydration solution to children with acute diarrhoea: A multicenter
European trail. Journal of Pediatric Gastroenterology and Nutrition, 30,
460.
Haller, D., Bode, C., Hammes, W. P., Pfeifer, A. M., Schiffrin, E. J., &
Blum, S. (2000). Non-pathogenic bacteria elicit a differential cytokine
response by intestinal epithelial cell/leucocyte co-cultures. Gut, 47,
7987.
He, F., Morita, H., Ouwehand, A. C., Hosoda, M., Hiramatsu, M.,
Kurisaki, J., et al. (2002). Stimulation of the secretion of pro-
inammatory cytokines by Bifidobacterium strains. Microbiology and
Immunology, 46, 781785.
Hessle, C., Hanson, L. A., & Wold, A. E. (1999). Lactobacilli from human
gastrointestinal mucosa are strong stimulators of IL-12 production.
Clinical and Experimental Immunology, 116, 276282.
Hilton, E., Kolakowski, P., Singer, C., & Smith, M. (1997). Efcacy of
Lactobacillus GG as a diarrhoeal preventive in Travellers. Journal of
Travel Medicine, 4, 4143.
Holm, F. (2003). Gut health and diet: The benets of probiotic and
prebiotics on human health. The World of Ingredients, 2, 5255.
Hosoda, M., Hashimoto, H., Morita, H., Chiba, M., & Hosono, A.
(1992). Antimutagenicity of milk cultured with lactic acid bacteria
against N-methyl-N-nitro-N-nitrosoguanidine. Journal of Dairy
Science, 75, 976981.
Hsieh, M. L., & Chou, C. C. (2006). Mutagenicity and antimutagenic
effect of soymilk fermented with lactic acid bacteria and bidobacteria.
International Journal of Food Microbiology, 111, 4347.
Hunter, J. O., Lee, A. J., King, T. S., Barratt, M. E. J., Linggood, M. A.,
& Blades, J. A. (1996). Enterococcus faecium strain PR88an effective
probiotic. Gut, 38(Suppl 1), A62.
Ji, S. T. (1997). Untersuchungen zur Klarung der Mechanismen bei
antigenotoxischen und antikanzerogenen Wirkungen durch Milchsaur-
ebakterien: Charakterisierung von bakteriellen Zellfraktionen fur die
Entgiftung von lebensmittelrelevanten Kanzerogenen. (Studies on the
antigenotoxic and anticarcinogenic effects of lactic acid bacteria:
characterization of bacterial cell fractions on detoxifying food relevant
carcinogens). Heidelberg, Germany: Medizinische Fakultat der Uni-
versitat (in German).
Kasper, H. (1996). Lebendkeime in fermentierten milchproduktenIhre
bedeutung fur die prophylaxe und therapie (Living bacteria in
fermented milk productsTheir importance for prophylaxis and
therapy). Ernahrungsumschau, 43, 4045 (in German).
Katelaris, P. H., Salam, I., & Farthing, M. J. (1995). Lactobacilli to
prevent travellers diarrhoea? New England Journal of Medicine, 333,
13601361.
Kato, I., Tanaka, K., & Yokokura, T. (1999). Lactic acid bacterium
potently induces the production of interleukin-12 and interferon-
gamma by mouse splenocytes. International Journal of Immunophar-
macology, 21, 121131.
Kawase, M., Hoshimoto, H., Hosoda, M., Morita, H., & Hosono, A.
(1999). Effect of administration of fermented milk containing whey
protein concentrate to rats and healthy men on serum lipids and blood
pressure. Journal of Dairy Science, 83, 255263.
Kiebling, G., Schneider, J., & Jahreis, G. (2002). Long-term consumption
of fermented dairy products over 6 months increases HDL cholesterol.
European Journal of Clinical Nutrition, 56, 843849.
Klaver, F. A. M., & Meer, R. V. D. (1993). The assumed assimilation of
cholesterol by lactobacilli and Bifidobacterium bifidum is due to their
bile salt deconjugating activity. Applied and Environmental Microbiol-
ogy, 59, 11201124.
ournal 17 (2007) 12621277Krishnakumar, V., & Gordon, I. R. (2001). Probiotics: Challenges and
opportunities. Dairy Industries International, 66, 3840.
-
ARTICLE IN PRESSiry JKurmann, J. A., & Rasic, J. L. (1991). The health potential of products
containing bidobacteria. In R. K. Robinson (Ed.), Therapeutic
properties of fermented milks (pp. 117157). London, UK: Elsevier
Applied Science Publishers.
Lankaputhra, W. E. V., & Shah, N. P. (1995). Survival of Lactobacillus
acidophilus and Bifidobacterium spp. in the presence of acid and bile
salts. Cultured Dairy Products Journal, 30, 27.
Lankaputhra, W. E. V., & Shah, N. P. (1997). Improving viability of
Lacidophilus and bidobacteria in yoghurt using two step fermentation
and neutralised mix. Food Australia, 49, 363369.
Lankaputhra, W. E. V., & Shah, N. P. (1998). Antimutagenic properties of
probiotic bacteria and of organic acids. Mutation Research, 39,
169182.
Le Leu, R. K., Brown, I. L., Hu, Y., Bird, A. R., Jackson, M., Esterman,
A., et al. (2005). A synbiotic combination of resistant starch and
Bifidobacterium lactis facilitates apoptotic deletion of carcinogen-
damaged cells in rat colon. Journal of Nutrition, 135, 9961001.
Leblanc, J. G., Matar, C., Valdez, J. C., Leblanc, J., & Perdigon, G.
(2002). Immunomodulating effects of peptidic fractions issued from
milk fermented with Lactobacillus helveticus. Journal of Dairy Science,
85, 27332742.
Lionetti, E., Miniello, V. L., Castellaneta, S. P., Magista, A. M., De canio,
A., Maurogiovanni, G., et al. (2006). Lactobacillus reuteri therapy to
reduce side-effects during anti-Helicobacter pylori treatment in
children: A randomized placebo controlled trial. Alimentary Pharma-
cology and Therapeutics, 24, 14611468.
Liong, M. T., & Shah, N. P. (2005). Acid and bile tolerance and
cholesterol removal ability of Lactobacilli strains. Journal of Dairy
Science, 88, 5566.
Liong, M. T., & Shah, N. P. (2006). Effects of Lactobacillus casei synbiotic
on serum lipoprotein, intestinal microora, and organic acids in rats.
Journal of Dairy Science, 89, 13901399.
MacFarlane, G. T., & Cummings, J. H. (2002). Probiotics, infection and
immunity. Current Opinion in Infectious Diseases, 15, 501506.
Marteau, P. R., De Vrese, M., Cellier, C. J., & Schrezenmeir, J. (2001).
Protection from gastrointestinal diseases with the use of probiotics.
American Journal of Clinical Nutrition, 73, 430S436S.
Marteau, P., Vaerman, J. P., Bord, J. P., Brassart, D., Pochart, P.,
Desjeux, J. F., et al. (1997). Effects of intrajejunal perfusion and
chronic ingestion of Lactobacillus johnsonii strain LA1 on serum
concentrations and jejunal secretions of immunoglobulins and serum
proteins in healthy humans. Gastroenterologie Clinique et Biologique,
21, 293298.
Matar, C., Amiot, J., Savoie, L., & Goulet, J. (1996). The effect of
milk fermentation by Lactobacillus helveticus on the release of
peptides during in vitro digestion. Journal of Dairy Science, 79,
971979.
Matar, C., Nadathur, S. S., Bakalinsky, A. T., & Goulet, J. (1997).
Antimutagenic effects of milk fermented by Lactobacillus helveticus
L89 and a protease-decient derivative. Journal of Dairy Science, 80,
19651970.
Matar, C., Valdez, J. C., Medina, M., Rachid, M., & Perdigon, G. (2001).
Immunomodulating effects of milks fermented by Lactobacillus
helveticus and its non-proteolytic variant. Journal of Dairy Research,
68, 601609.
Matsuzaki, T. (1998). Immunomodulation by treatment with Lactobacillus
casei strain Shirota. International Journal of Food Microbiology, 41,
133140.
Mccracken, V. J., Chun, T., Baldeon, M. E., Ahrne, S., Molin, G.,
Mackie, R. I., et al. (2002). TNF-alpha sensitizesHT-29 colonic
epithelial cells to intestinal lactobacilli. Experimental Biology and
Medicine, 227, 665670.
McFarland, L. V. (2006). Meta-analysis of probiotics for the prevention of
antibiotic associated diarrhea and the treatment of Clostridium difficile
disease. American Journal of Gastroenterology, 101, 812822.
Metchnikoff, L. E. (2004). The prolongation of life: Optimistic studies
N.P. Shah / International Da(reprinted edition 1907). New York, USA: Springer Publishing
Company.Miettinen, M., Vuopio-Varkila, J., & Varkila, K. (1996). Production
of human tumor necrosis factor alpha, interleukin-6, and interleukin-
10 is induced by lactic acid bacteria. Infection and Immunity, 64,
54035405.
Morita, H., He, F., Fuse, T., Ouwehand, A. C., Hashimoto, H., Hosoda,
M., et al. (2002). Cytokine production by the murine macrophage cell
line J774.1 after exposure to lactobacilli. Bioscience, Biotechnology,
Biochemistry, 66, 19631966.
Morotomi, M., & Mutai, M. (1986). In vitro binding of potent mutagenic
pyrolysates. Journal of National Cancer Institute, 77, 195201.
Moschner, O., Daniela, L., Gerhard, J., Gerhard, R., & Pool-Zobel, B. L.
(2004). Dietary intervention with the probiotics Lactobacillus acid-
ophilus 145 and Bifidobacterium longum 913 modulates the potential of
human faecal water to induce damage in HT29clone19A cells. British
Journal of Nutrition, 91(6), 925932.
Mrda, Z., Zivanovic, M., Rasic, J., Gajin, S., Somer, L., & Trbojevic, S.
(1998). Therapy of Helicobacter pylori infection using Lactobacillus
acidophilus. Meditsinski Pregled, 51, 343345.
Mukai, T., Asasaka, T., Sato, E., Mori, K., Matsumoto, M., & Ohori, H.
(2002). Inhibition of binding of Helicobacter pylori to the glycolipid
receptors by probiotic Lactobacillus reuteri. FEMS Immunology and
Medical Microbiology, 32, 105110.
Myllyluoma, E., Veijola, L., Ahlroos, T., Tynkkynen, S., Kankuri, E.,
Vapaatalo, H., et al. (2005). Probiotic supplementation improves
tolerance to Helicobacter pylori eradication therapyA placebo-
controlled, double-blind randomized pilot study. Alimentary Pharma-
cology and Therapeutics, 21, 12631272.
Nakamura, Y., Yamamoto, N., Sakai, K., Okubo, A., Yamazaki, S., &
Takano, T. (1995). Purication and characterization of angiotensin
I-converting enzyme inhibitors from a sour milk. Journal of Dairy
Science, 78, 777783.
Nista, E. C., Candelli, M., Cremonini, F., Cazzato, I. A., Zocco, M. A.,
Franceschi, F., et al. (2004). Bacillus clausii therapy to reduce side-
effects of anti-Helicobacter pylori treatment: Randomized, double-
blind, placebo controlled trial. Alimentary Pharmacology and Ther-
apeutics, 20, 11811188.
Nobaek, S., Johansson, M. L., & Molin, G. (2000). Alteration of intestinal
microora is associated with reduction in abdominal bloating and pain
in patients with irritable bowel syndrome. American Journal of
Gastroenterology, 95, 12311238.
Ong, L., Henriksson, H., & Shah, N. P. (2006). Development of probiotic
Cheddar cheese containing Lactobacillus acidophilus, Lb. casei, Lb.
paracasei and Bifidobacterium spp. and their inuence on proteolytic
patterns and production of organic acid. International Dairy Journal,
16, 446456.
Orrhage, K., Sillerstrom, E., Gustafsson, J. A., Nord, C. E., & Rafter, J.
(1994). Binding of mutagenic heterocyclic amines by intestinal and
lactic acid bacteria. Mutation Research, 311, 239248.
Ouwehand, A. C., Salminen, S., & Isolauri, E. (2002). Probiotics: An
overview of benecial effects. Antonie Van Leeuwenhoek, 82, 279289.
Pereira, D. I. A., & Gibson, G. R. (2002). Cholesterol assimilation by
lactic acid bacteria and bidobacteria isolated from the human gut.
Applied and Environmental Microbiology, 68, 46894693.
Pinchuk, I. V., Bressollier, P., Verneuil, B., Fenet, B., Sorokulova, I. B.,
Me0graud, F., et al. (2001). In vitro anti-Helicobacter pylori activity ofthe probiotic strain Bacillus subtilis 3 is due to secretion of antibiotics.
Antimicrobial Agents and Chemotherapy, 45, 31563161.
Playne, M. J., Bennet, L. E., & Smithers, G. W. (2003). Functional dairy
foods and ingredients. The Australian Journal of Dairy Technology, 58,
242264.
Pochapin, M. (2000). The effect of probiotics on Clostridium difficile
diarrhea. American Journal of Gastroenterology, 95, S11S13.
Pool-Zobel, B. L., Bertram, B., Knoll, M., Lambertz, R., Neudecker, C.,
Schillinger, U., et al. (1993). Antigenotoxic properties of lactic acid
bacteria in vivo in the gastrointestinal tract of rats. Nutrition and
Cancer, 20(3), 271281.
ournal 17 (2007) 12621277 1275Pool-Zobel, B. L., Neudecker, C., Domizlaff, I., Ji, S., Schillinger, U.,
Rumney, C., et al. (1996). Lactobacillus and Bifidobacterium mediated
-
ARTICLE IN PRESSry Jantigenotoxicity in the colon of rats. Nutrition and Cancer, 26,
365380.
Pool-Zobel, B. L., Bertram, B., & Knoll, M. (1993). Antigenotoxic
properties of lactic acid bacteria in vivo in the gastrointestinal tract of
rats. Nutrition and Cancer, 20, 271282.
Rangavajhyala, N., Shahani, K. M., Sridevi, G., & Srikumaran, S. (1997).
Nonlipopolysaccharide component(s) of Lactobacillus acidophilus
stimulate(s) the production of interleukin-1 alpha and tumor necrosis
factor-alpha by murine macrophages. Nutrition and Cancer, 28,
130134.
Richelsen, B., Kristensen, K., & Pedersen, S. B. (1996). Long-term (6
months) effect of a new fermented milk product on the level of plasma
lipoproteinsA placebo-controlled and double blind study. European
Journal of Clinical Nutrition, 50, 811815.
Saavedra, J. M., Bauman, N. A., Oung, I., Perman, J. A., & Yolken, R. H.
(1994). Feeding of Bifidobacterium bifidum and Streptococcus thermo-
philus to infants in hospital for prevention of diarrhoea and shedding
of rotavirus. Lancet, 344, 10461049.
Saint Marc, T., Rosse, H. O., Prats, L., & Touraine, J. L. (1999). Efcacit
Saccharomyces boulardii dans let treatment des diarrheas di SIDA.
Annals of Internal Medicine, 142, 6465.
Sakamoto, S., Igarashi, M., Kimura, K., Takagi, A., Miwa, T., & Koga,
Y. (2001). Journal of Antimicrobial Chemistry, 47, 709710.
Sanders, M. E., & Klaenhammer, T. R. (2001). Invited review: The
scientic basis of Lactobacillus acidophilus NCFM functionality as a
probiotic. Journal of Dairy Science, 84, 319331.
Schiffrin, E. J., Brassart, D., Servin, A. L., Rochat, F., & Donnet-Hughes,
A. (1997). Immune modulation of blood leukocytes in humans by
lactic acid bacteria: Criteria for strain selection. American Journal of
Clinical Nutrition, 66, 515S520S.
Schiffrin, E. J., Rochat, F., Link-Amster, H., Aeschlimann, J. M., &
Donnet-Hughes, A. (1995). Immunomodulation of human blood cells
following the ingestion of lactic acid bacteria. Journal of Dairy Science,
78, 491497.
Schultz, M., & Sartor, R. B. (2000). Probiotics and inammatory bowel
diseases. American Journal of Gastroenterology, 95, 19S21S.
Shah, N. P. (1993). Effectiveness of dairy products in alleviation of lactose
intolerance. Food Australia, 45, 268271.
Shah, N. P. (1999). Probiotic bacteria: Antimicrobial and antimutagenic
properties. Probiotica, 6, 13.
Shah, N. P. (2000a). Probiotic bacteria: Selective enumeration and
survival in dairy foods. Journal of Dairy Science, 83, 894907.
Shah, N. P. (2000b). Effects of milk-derived bioactives: An overview.
British Journal of Nutrition, 84, S3S10.
Shah, N. P. (2000c). Some benecial effects of probiotic bacteria.
Bioscience Microflora, 19, 99106.
Shah, N. P. (2001). Functional foods, probiotics and prebiotics. Food
Technology, 55, 4653.
Shah, N. P. (2003). Yogurt: The product and its manufacture. In L.
Trugo, P. Finglas, & B. Caballero (Eds.), Encyclopedia of food
sciences and nutrition (2nd ed., pp. 62526260). London, UK:
Academic Press.
Shah, N. P. (2004). Probiotics and prebiotics. AgroFood Industry HiTech,
15, 1316.
Shah, N. P. (2006a). Health benets of yogurt and fermented milks. In
R. C. Chandan (Ed.), Manufacturing yogurt and fermented milks
(pp. 327340). Iowa, USA: Blackwell Publishing Professional.
Shah, N. P. (2006b). Microorganisms and health attributes (probiotics).
In R. C. Chandan (Ed.), Manufacturing yogurt and fermented milks
(pp. 341354). Iowa, USA: Blackwell Publishing Professional.
Shah, N. P., Ali, J. F., & Ravula, R. R. (2000). Populations of L.
acidophilus, Bifidobacterium spp., and L. casei in commercial fermented
milk products. Bioscience and Microflora, 19, 3539.
Shah, N. P., Fedorak, R. N., & Jelen, P. (1992). Food consistency effects
of quarq in lactose absorption by lactose intolerant individuals.
International Dairy Journal, 2, 257269.
N.P. Shah / International Dai1276Shah, N. P., & Lankaputhra, W. E. V. (2002). Bifidobacterium spp.:
Morphology and physiology. In H. Roginski, J. Fuquay, P. Fox, &Patrick (Eds.), Encyclopedia of dairy science (pp. 141146). London,
UK: Academic Press.
Shah, N. P., Lankaputhra, W. E. V., Britz, M. L., & Kyle, W. S. A. (1995).
Survival of Lactobacillus acidophilus and Bifidobacterium bifidum in
commercial yoghurt during refrigerated storage. International Dairy
Journal, 5, 515521.
Sheu, B. S., Wu, J. J., Lo, C. Y., Wu, H. W., Chen, J. H., Lin, Y. S., et al.
(2002). Impact of supplement with Lactobacillus- and Bifidobacterium-
containing yogurt on triple therapy for Helicobacter pylori eradication.
Alimentary Pharmacology and Therapeutics, 16, 16691675.
Shornikova, A. V., Casas, I. A., & Isolauri, E. (1997b). Lactobacillus
reuteri as a therapeutic agent in acute diarrhea in young children.
Journal of Pediatric Gastroenterology and Nutrition, 24, 399404.
Shornikova, A. V., Isolauri, E., Burnakova, L., Lukovnikova, S., &
Vesikari, T. (1997a). A trial in the Karelian Republic of oral
rehydration and Lactobacillus GG for treatment of acute diarrhoea.
Acta Paediatrics, 86, 460465.
Singh, J., Rivenson, A., Tomita, M., Shimamura, S., Ishibashi, N., &
Reddy, B. S. (1997). Bifidobacterium longum and Lactobacillus
acidophilus producing intestinal bacteria inhibits colon cancer and
modulates the intermediate biomarkers of colon carcinogenesis.
Carcinogenesis, 18, 833841.
Sreekumar, O., & Hosono, A. (1998). Antimutagenicity and the inuence
of physical factors in binding Lactobacillus gasseri and Bifidobacterium
longum cells to amino acid pyrolysates. Journal of Dairy Science, 81,
15081516.
Tamime, A. Y., Saarela, M., Sondergaard, K., Mistry, V. V., & Shah, N.
P. (2005). Production and maintaining viability of probiotics (Chapter
3). In A. Y. Tamime (Ed.), Probiotic dairy products (pp. 3972).
Oxford, UK: Blackwell Publishing.
Taranto, M. P., Medici, M., Perdigon, G., Ruiz Holgado, A. P., & Valdez,
G. F. (1998). Evidence for hypocholesterolemic effect of Lactobacillus
reuteri in hypercholesterolemic mice. Journal of Dairy Science, 81,
23362340.
Tharmaraj, N., & Shah, N. P. (2004). Survival of Lactobacillus acidophilus,
Lactobacillus paracasei subsp. paracasei, Lactobacillus rhamnosus,
Bifidobacterium animalis and Propionibacterium in cheese-based dips
and the suitability of dips as effective carriers of probiotic bacteria.
International Dairy Journal, 14, 10551066.
Tomioka, H., Tomioka, K., Sato, K., & Saito, H. (1992). The protective
activity of immunostimulants against Listeria monocytogenes infection
in mice. Journal of Medical Microbiology, 36, 112116.
Tsai, C. C., Huang, L. F., Lin, C. C., & Tsen, H. Y. (2004). Antagonistic
activity against Helicobacter pylori infection in vitro by a strain
of Enterococcus faecium TM39. International Journal of Food Micro-
biology, 96, 112.
Tsangalis, D., Ashton, J., Mcgill, A., & Shah, N. P. (2002). Enzymic
transformation of isoavone phytoestrogens in soymilk by b-glucosi-dase producing bidobacteria. Journal of Food Science, 67(8),
31043113.
Tsangalis, D., Ashton, J. F., Wilcox, G., Shah, N. P., & Stojanovska, L.
(2005). Bioavailability of isoavone phytoestrogen in postmenopausal
women consuming soy milk fermented with probiotic bidobacteria.
British Journal of Nutrition, 93, 867877.
Usman, A., & Hosono, A. (2000). Effect of administration of Lactoba-
cillus gasseri on serum lipids and fecal steroid