www.elsevier.com/locate/jbiotec
Review article
Received 19 July 2000; received in revised form 21 September 2000; accepted 4 October 2000
Abstract
During the past two decades probiotic (health promoting) micro-organisms have been increasingly included in
various types of food products, especially in fermented milks. Several aspects, including safety, functional and
technological characteristics, have to be taken into consideration in the selection process of probiotic micro-organisms. Safety aspects include specifications such as origin (healthy human GI-tract), non-pathogenicity and antibiotic
resistance characteristics. Functional aspects include viability and persistence in the GI-tract, immunomodulation,
antagonistic and antimutagenic properties. Before probiotic strains, chosen on the basis of their good safety and
functional characteristics, can benefit the consumer, they must first be able to be manufactured under industrial
conditions. Furthermore, they have to survive and retain their functionality during storage, and also in the foods into
which they are incorporated without producing off-flavours. Factors related to the technological and sensory aspects
of probiotic food production are of utmost importance since only by satisfying the demands of the consumer can the
food industry succeed in promoting the consumption of functional probiotic products in the future. 2000 Elsevier
Science B.V. All rights reserved.
Keywords: Probiotic bacteria; Health; Safety
1. Introduction
0168-1656/00/$ - see front matter 2000 Elsevier Science B.V. All rights reserved.
PII: S 0 1 6 8 - 1 6 5 6 ( 0 0 ) 0 0 3 7 5 - 8
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assess their health benefits to consumers (MattilaSandholm and Salminen, 1998; Mattila-Sandholm
et al., 1999).
Fig. 1. The theoretical basis for selection of probiotic microorganisms includes safety, functional (survival, adherence,
colonisation, antimicrobial production, immune stimulation,
antigenotoxic activity and prevention of pathogens) and technological aspects (growth in milk, sensory properties, stability,
phage resistance, viability in processes).
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200
target precursor for vancomycin activity terminating with D-alanine (Billot-Klein et al., 1994).
Many intrinsically vancomycin resistant strains of
lactobacilli have a long history of safe use as
probiotics and there is no indication that vancomycin resistant lactobacilli could transfer the
resistance to other bacteria. Tynkkynen et al.
(1998) have demonstrated that the vancomycin
resistance factor of the probiotic strain L. rhamnosus GG is not closely related to those of enterococci, and they could not observe the transfer of
antibiotic resistances between L. rhamnosus GG
and enterococci.
Although plasmid-linked antibiotic resistances
are not very common among lactobacilli, they do
occur (Ishiwa and Iwata, 1980; Vescovo et al.,
1982; Rinckel and Savage, 1990), and their safety
implications should be taken into consideration.
Since transfer of antibiotic resistant genes may
occur between phylogenetically distant bacteria
(Courvalin, 1994), strains harbouring mobile elements carrying resistance genes should not be
used either as human or animal probiotics.
Checking the ability of a proposed probiotic
strain to act as a donor of conjugative antibiotic
resistance genes may be a prudent precaution
especially when probiotics are administered during antibiotic therapy, and at least in the case of
animal feeding, where the use of antibiotics as
growth promoters apparently creates selective advantage for spreading of the resistance factors.
Antibiotic susceptibility patterns vary greatly between different species of lactobacilli, and strains
with atypical resistance to some clinically important antibiotics have been detected among lactobacilli (Charteris et al., 1998b; Felten et al., 1999),
indicating the necessity for susceptibility testing of
each probiotic strain.
Most bifidobacteria are intrinsically resistant to
nalidixic
acid,
neomycin,
polymyxin
B,
kanamycin, gentamycin, streptomycin and
metronidazole (Miller and Finegold, 1967; Matteuzzi et al., 1983; Charteris et al., 1998a). In
earlier studies vancomycin has been found highly
inhibitory against bifidobacteria (Miller and Finegold, 1967; Lim et al., 1993), whereas in a recent
study by Charteris et al. (1998a) vancomycin resistance was suggested to be a general characteris-
201
Saccharomyces boulardii
Strain
Table 1
Clinical effects of some probiotic strains and yoghurt strains
References
202
M. Saarela et al. / Journal of Biotechnology 84 (2000) 197215
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204
with transient alterations beneficial to the consumer (Schiffrin et al., 1997; Mattila-Sandholm
and Kauppila, 1998). L. rhamnosus GG and
Bifidobacterium lactis Bb-12 derived extracts have
been shown to suppress lymphocyte proliferation
in vitro (Mattila-Sandholm and Kauppila, 1998).
Further evidence for immunomodulation by these
two strains was provided by a trial involving
children with severe atopic eczema resulting from
food allergy. Children fed L. rhamnosus GG and
B. lactis Bb-12 showed a significant improvement
in clinical symptoms compared to the placebo
group (Mattila-Sandholm and Kauppila, 1998;
Alander and Mattila-Sandholm, 2000). L. rhamnosus GG has been shown to down-regulate the
milk-induced inflammatory response in milk-hypersensitive subjects, whereas it had an immunostimulatory effect in healthy subjects (Pelto et al.,
1998). Furthermore, L. rhamnosus GG was shown
to promote IgA immune response in Crohns disease patients (Malin et al., 1996) and enhanced
the circulating antibody secreting cell response in
children with rotavirus diarrhoea (Kaila et al.,
1992).
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Main flavour
components
Lactobacillus acidophilus a
Bifidobacterium spp.a
Streptococcus thermophilus b
Lactate (DL)
Lactate (L+), acetate
Lactate (L+),
acetaldehyde, diacetyl
Lactate (D),
acetaldehyde, diacetyl
Probiotic strains.
Non-probiotic strains.
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was ten times higher with the sweet milk (Pettersson et al., 1983).
6. Future trends
Diet is a major focus of public health strategy
aimed at maintaining optimum health throughout
life, preventing early onset of chronic diseases
such as gastrointestinal disorders, cardiovascular
disease, cancer, osteoporosis, as well as promoting
healthier ageing. Although the highly complex
relationship of food and health is still poorly
understood, recent research advances in different
disciplines provide promising new approaches to
improve our understanding. The growing demand
for healthy foods is stimulating innovation and
new product development in the food industry
internationally. The food industry has a central
role in facilitating healthier eating practices
through the provision and promotion of healthy
foods.
Continuously increasing consumer health consciousness and expenditure are socio-economic
factors responsible for the expanding European
and world-wide interest in functional foods. Considerable confusion and scepticism, however, exists between consumers, consumer organisations,
scientific communities and media about the claims
associated with probiotic products. Recent EU
projects have demonstrated that, with co-ordinated efforts towards communication and a scientific approach to selecting and applying
probiotics, functional food products can be developed with measurable health benefits for consumers. Probiotic strains can be successfully
manufactured and incorporated into highly acceptable food products where they can retain their
viability and functionality. There are many strain
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Acknowledgements
This work was supported by European Concerted Action PL98 4230.
References
Adams, M.R., 1999. Safety of industrial lactic acid bacteria. J.
Biotechnol. 68, 171 178.
210
glycopeptide-resistant species Lactobacillus casei, Pediococcus pentosaceus, Leuconostoc mesenteroides, and Enterococcus gallinarum. J. Bacteriol. 176, 2398 2405.
Black, F., Einarsson, K., Lidbeck, A., Orrhage, K., Nord,
C.E., 1991. Effect of lactic acid producing bacteria on the
human intestinal microflora during ampicillin treatment.
Scand. J. Infect. Dis. 23, 247 254.
Black, F.T., Andersen, P.L., Orskov, J., Orskov, F., Gaarslev,
K., Laulund, S., 1989. Prophylactic efficacy of lactobacilli
on travellers diarrhoea. Travel Med. 333 335.
Bleichner, G., Blehaut, H., Mentec, H., Moyse, D., 1997.
Saccharomyces boulardii prevents diarrhea in critically ill
tube-fed patients. A multicenter, randomized, double-blind
placebo-controlled trial. Intensive Care Med. 23, 517 523.
Buts, J.P., Corthier, G., Delmee, M., 1993. Saccharomyces
boulardii for Clostridium difficile-associated enteropathies
in infants. J. Pediatr. Gastroenterol. Nutr. 16, 419 425.
Charteris, W.P., Kelly, P.M., Morelli, L., Collins, J.K., 1998a.
Antibiotic susceptibility of potentially probiotic Bifidobacterium isolates from the human gastrointestinal tract. Lett.
Appl. Microbiol. 26, 333 337.
Charteris, W.P., Kelly, P.M., Morelli, L., Collins, J.K., 1998b.
Antibiotic susceptibility of potentially probiotic Lactobacillus species. J. Food Prot. 61, 1636 1643.
Chauviere, G., Cocoinnier, M.-H., Kerneis, S., Fourniat, J.,
Servin, A.L., 1992. Adhesion of human Lactobacillus
acidophilus strain LB to human enterocyte-like Caco-2
cells. J. Gen. Microbiol. 138, 1689 1696.
Coconnier, M.-H., Bernet, M.-F., Chauviere, G., Servin, A.L.,
1993a. Adhering heat-killed human Lactobacillus
acidophilus, strain LB, inhibits the process of pathogenicity
of diarrhoeagenic bacteria in cultured human intestinal
cells. J. Diarrh. Dis. 11, 235 242.
Coconnier, M.-H., Bernet, M.-F., Kerneis, S., Chauviere, G.,
Fourniat, J., Servin, A.L., 1993b. Inhibition of adhesion of
enteroinvasive pathogens to human intestinal Caco-2 cells
by Lactobacillus acidophilus strain LB decreases bacterial
invasion. FEMS Microbiol. Lett. 110, 299 306.
Coconnier, M.-H., Lievin, V., Bernet-Camard, M.-F., Hudault, S., Servin, A.L., 1997. Antibacterial effect of the
adhering human Lactobacillus acidophilus strain LB. Antimicrob. Agents Chemother. 41, 1046 1052.
Courvalin, P., 1994. Transfer of antibiotic resistance genes
between gram-positive and gram-negative bacteria. Antimicrob. Agents Chemother. 38, 1447 1451.
Daly, C., Davis, R., 1998. The biotechnology of lactic acid
bacteria with emphasis on applications in food safety and
human health. Agric. Food Sci. Finland 7, 219 250.
Dave, R., Shah, N., 1997. Viability of yoghurt and bacteria in
yoghurts made from commercial starter cultures. Int.
Dairy J. 7, 31 41.
Dave, R.I., Shah, N.P., 1998. Ingredient supplementation effects on viability of probiotic bacteria in yougurt. J. Dairy
Sci. 81, 2804 2816.
Donnet-Hughes, A., Rochat, F., Serrant, P., Aeschlimann,
J.M., Schiffrin, J.E., 1999. Modulation of nonspecific
mechanisms of defense by lactic acid bacteria: effective
dose. J. Dairy Sci. 82, 863 869.
211
212
Lim, K.S., Huh, C.S., Baek, Y.J., 1993. Antimicrobial susceptibility of bifidobacteria. J. Dairy Sci. 76, 2168 2174.
Ling, W.H., Korpela, R., Mykkanen, H., Salminen, S., Hanninen, O., 1994. Lactobacillus strain GG supplementation
decreases colonic hydrolytic and reductive enzyme activities in healthy female adults. J. Nutr. 124, 18 23.
Link-Amster, H., Rochat, F., Saudan, K.Y., Mignot, O.,
Aeschlimann, J.M., 1994. Modulation of a specific humoral immune response and changes in intestinal flora
mediated through fermented milk intake. FEMS Immunol.
Med. Microbiol. 10, 55 63.
Mackay, A.D., Taylor, M.B., Kibbler, C.C., Hamilton-Miller,
J.M.T., 1999. Lactobacillus endocarditis caused by a probiotic organism. Clin. Microbiol. Infect. 5, 290 292.
Majamaa, H., Isolauri, E., 1997. Probiotics: a novel approach
in the management of food allergy. J. Allergy Clin. Immunol. 99, 179 185.
Majamaa, H., Isolauri, E., Saxelin, M., Vesikari, T., 1995.
Lactic acid bacteria in the treatment of acute rotavirus
gastroenteritis. J. Pediatr. Gastroenterol. Nutr. 20, 333
338.
Malin, M., Suomalainen, H., Saxelin, M., Isolauri, E., 1996.
Promotion of IgA immune response in patients with
Crohns disease by oral bacteriotherapy with Lactobacillus
GG. Ann. Nutr. Metab. 40, 137 145.
Marteau, P., Pochart, P., Bouhnik, Y., Rambaud, J.C., 1993.
Fate and effects of some transiting micro-organisms in the
human gastrointestinal tract. World Rev. Nutr. Diet. 74,
1 21.
Marteau, P., Gerhardt, M.F., Myara, A., Bouvier, E., Trivin,
F., Rambaud, J.C., 1995. Metabolism of bile salts by
alimentary bacteria during transit in the human small
intestine. Microb. Ecol. Health Dis. 8, 151 157.
Marteau, P., Pochart, P., Flourie, B., Pellier, P., Santos, L.,
Desjeux, J.F., Rambaud, J.C., 1990. Effect of chronic
ingestion of a fermented dairy product containing Lactobacillus acidophilus and Bifidobacterium bifidum on
metabolic activities of the colonic flora in humans. Am. J.
Clin. Nutr. 52, 685 688.
Marteau, P., Vaerman, J.P., Dehennin, J.P., Bord, S., Brassart, D., Pochart, P., Desjeux, J.F., Rambaud, J.C., 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. Gastroenterol. Clin. Biol. 21,
293 298.
Matteuzzi, D., Crociani, F., Brigidi, P., 1983. Antimicrobial
susceptibility of Bifidobacterium. Ann. Microbiol. (Paris)
134A, 339 349.
Mattila-Sandholm, T., Kauppila, T. (Eds.), 1998. Functional
Food Research in Europe 3rd Workshop, FAIR CT961028, PROBDEMO, VTT Symposium 187. Haikko, Finland, p. 125.
Mattila-Sandholm, T., Salminen, S., 1998. Up-to-date on probiotics in Europe. Gastroenterol. Int. 11, 8 16.
Mattila-Sandholm, T., Matto, J., Saarela, M., 1999. Lactic
acid bacteria with health claims interference and interactions with gastrointestinal flora. Int. Dairy J. 9, 25 35.
213
214
215