Available online at www.sciencedirect.

com

Application of probiotics in food products—challenges and

new approaches
I Jankovic1, W Sybesma2, P Phothirath1, E Ananta1 and A Mercenier1

The probiotic research conducted over the past 20 years has bacteria and thereby reduce putrefaction in the gut, thus
resulted in a valuable source of data related to health beneficial prolonging the life span of the host. Soon after his
effects of probiotics. Nevertheless, documentation of probiotic postulate, strains of LAB and bifidobacteria were applied
benefits remains challenging, especially in functional foods that as supplements and over-the counter drugs for treatment
are designed for the generally healthy population that, however, of diarrhea (e.g. Lactobacillus LB Lactéol in 1907, Escher-
regularly experiences episodes of ‘suboptimal’ health. In ichia coli Nissle 1917, end of 1920s) and in food products
addition, in view of today’s application of probiotics in an for promotion of intestinal health and prevention of
increasing variety of food matrixes, process optimization and disease (e.g. Lactobacillus acidophilus L-92 in 1910, Lacto-
product design need to take into account cell viability and bacillus casei Shirota in 1935). Note that two of these early
probiotic function altogether. To meet this challenge, medium products (Lactéol and L-92) were heat inactivated. Pro-
to high-throughput bioassays – based on the identification of biotics were also introduced in animal feed in the 1970s as
active compounds and their mechanism of action – have to be supplements for the promotion of animal growth and
developed and their predictive value established. Together with improvement of their resistance to disease [3]. Approxi-
validated biomarkers for health and disease, this should help mately one century after the initial probiotic concept, the
rationalize probiotic product development and associated transition from theory to scientific documentation of
health claim substantiation in human studies. beneficial effects of probiotics had begun. Apart from
Addresses some pioneering work in the early days, concerted efforts
1
Nestlé Research Center, Vers-chez-les-Blanc, 1000 Lausanne 26, to demonstrate health beneficial effects of probiotics
Switzerland mainly started in the 1980s. The volume of research
2
Nestlé Product Technology Centre Konolfingen, Konolfingen,
Switzerland
rapidly accelerated after the year 2000 such that to date,
more than 700 human intervention trials have been con-
Corresponding author: Jankovic, I ([email protected]) ducted. In solely 2008, more than 1000 articles and
reviews were published on the subject and more than
Current Opinion in Biotechnology 2010, 21:175–181
2000 probiotic products launched (Figure 1).

This review comes from a themed issue on The aim of this paper is to review the probiotic devel-
Food biotechnology opment in the past years as well as the current challenges
Edited by Dietrich Knorr and Carmen Wacher
of using probiotics in different food matrixes.
Available online 21st April 2010

0958-1669/$ – see front matter Health beneficial effects of probiotics

# 2010 Elsevier Ltd. All rights reserved. The role of probiotics as functional ingredients in food
Generally, throughout life healthy individuals suffer
DOI 10.1016/j.copbio.2010.03.009
from periods of suboptimal health. This can be caused
by respiratory or gut infections or by other external
stimuli that challenge the immune system [4], and often
Introduction from first hypothesis to scientific alter the microbiota and weaken mucosal barrier func-
approach tion. A suboptimal state of health can also be a con-
The health-promoting use of fermented milk products sequence of chronic stresses, such as chronic infections,
started a long time before the existence of microorgan- fatigue, exercise, use of medications, psychological stres-
isms and lactic acid bacteria was discovered. The early ses, and many other challenges resulting in downregula-
written records go back to 76 B.C. when Roman historian tion or weakening of the natural defenses of the organism
Plinio (Plinius) described their use in the therapy of [4,5]. It is also well established that psychological stresses
various gastro-intestinal infections [1]. However, it was like depression or anxiety can influence digestive func-
only after the invention of the microscope and the dis- tion and symptom perception [6] thereby resulting in gut
covery of bacteria and especially lactic acid bacteria discomfort.
(LAB) in the 17th, 18th, and 19th centuries, respectively,
that the scientific basis of the probiotic concept was set The role of functional foods is to benefit human health
with the theories of Elie Metchnikoff at the beginning of beyond the effect of nutrients [7]. Situated between
the 20th century [2]. He postulated that consumption of foods, which supply basic physiological functions and
fermented milk would suppress the growth of proteolytic drugs that treat diseases, functional foods are used to

www.sciencedirect.com Current Opinion in Biotechnology 2010, 21:175–181

176 Food biotechnology

Figure 1

Publications and probiotic products. In gray: number of publications on probiotics published per year (source PubMed, http://www.ncbi.nlm.nih.gov/
sites/entrez). In black: number of products containing probiotics launched per year (source Global New Products Database, http://www.gnpd.com).

maintain good health and counterbalance small physio- usually interpreted as an increase in lactobacilli and/or
logical disorders that healthy hosts may experience. In bifidobacteria and a decrease in potentially pathogenic
addition to the well-established functional ingredients bacteria. In the past 20 years, it has been demonstrated
such as vitamins, minerals, and micronutrients, probiotics that it is possible to transiently modify the composition of
belong to the emerging generation of active ingredients the gut microbiota of healthy individuals in favor of
that includes prebiotics, phytonutrients, and lipids, for lactobacilli and bifidobacteria species upon ingestion of
example. some probiotics [8]. It was also shown that infants fed
probiotic infant formula have similar fecal levels of bifi-
Historically, the development of probiotics was very dobacteria as breast fed infants [9]. However, it remains
much oriented toward pharmaceutical applications such difficult to link such changes with a benefit in healthy
as treatment of diarrhea, prevention of antibiotic-associ- populations, although it is well established that dysbioses
ated diarrhea, management of stomach and gastro-intes- are associated to conditions such as chronic inflammatory
tinal infections, management of chronic inflammation, disorders [5], obesity [10], or allergy [11]. One of the best
and so on. However, these effects are not easily extended studied examples of how microbiota dysbiosis affects
to the category of functional foods that are destined for health is seen in Crohn’s disease (CD). A decrease in
the generally healthy population. Although beneficial the global biodiversity of intestinal bacteria, particularly
effects of specific probiotics have been demonstrated within the phylum Firmicutes, has been observed in CD
in the treatment and prevention of several health dis- patients [5]. Recent analyses have revealed that a lower
orders, the remaining challenge is to demonstrate long- level of Faecalibacterium prausnitzii, a major member of
term effects of probiotic foods as presently required by Firmicutes is associated with a higher risk of postoper-
health claim regulations in Europe. As large trials of long ative recurrence of ileal CD. Oral administration of live F.
duration are difficult to support, particularly for small and prausnitzii or its culture supernatant reduced the severity
medium-sized laboratories and food companies, there is of TNBS induced colitis in mice indicating that counter-
an urgent need to better identify and validate risk factors balancing dysbiosis might be a promising strategy in CD
of diseases and biomarkers of health. treatment [12].

Probiotics and microbiota balance Although it is not known whether alteration in the micro-
Microbiota balance is the oldest proposed probiotic biota is a cause or a consequence of a pathophysiological
benefit. Metchnikoff defined it as ‘seeding’ of the intes- situation, the aforementioned examples underline the
tinal tract with harmless LAB that suppress the growth of fact that an equilibrated microbiota is of high importance
harmful proteolytic bacteria. Nowadays such a benefit is for health maintenance. However, the challenge will be to

Current Opinion in Biotechnology 2010, 21:175–181 www.sciencedirect.com

 Probiotics for food Jankovic et al. 177

demonstrate i) what is the composition and function of a greatly help in proving the benefit of probiotic foods for
balanced microbiota and ii) its long-term impact on the healthy population as well as in validating immune
health. Efforts are currently ongoing to determine the markers.
composition and functionality of the microbiota in
healthy and diseased populations. A systems biology Intestinal discomfort
approach using techniques such as high-throughput Periods of gastro-intestinal discomfort are very frequent in
microbiota diversity diagnostic arrays [13], metage- otherwise healthy individuals. Irritable bowel syndrome
nomics (http://www.metahit.eu) [14], proteomics [15], (IBS), one of the most common disorders seen by primary
metabonomics [16], and high-throughput phenotyping care physicians, affects 7–10% of the world population [32].
of metagenomic clones [17] will progressively provide a In the absence of an efficient therapy with no side effects,
better definition of the composition and function of a well selected probiotic strains might provide a valuable
‘healthy’ microbiota. Markers of ‘healthy’ microbiota will alternative. Certainly, a significant reduction in IBS symp-
permit to definitively support Metchnikoff’s theory of toms has been observed after intervention with probiotics
‘balanced microbiota’ and its impact on health. such as B. infantis 35624 [33] or with a probiotic mix [34].
As another example of intestinal discomfort, constipation
Probiotics and immune system that represents the most prevalent complaint among the
Since the early studies of mucosal immunity in the 1970s, general adult population was shown to be regulated by a
a lot of progress has been made in understanding the probiotic fermented milk containing B. animalis DN-
mode of interaction between the gut microbiota and the 173010 [35,36]. Finally, successful treatment of colicky
immune system (e.g. see review [18]). The ability of symptoms, frequent in newborn infants, was achieved
exogenous probiotics to improve clinical outcomes using probiotic L. reuteri ATCC55730 [37].
through modulation of the immune response has been
demonstrated in subjects with chronic and acute diseases. Probiotic effects beyond the gut
For example, the probiotic mix VSL#3 was shown to It is important to note that a positive impact of probiotics
reduce pouchitis relapse [19,20] and to improve clinical is progressively being demonstrated beyond the gut. To
scores in ulcerative colitis patients [21,22] through im- cite only a few examples, oral intake of Lactobacillus
provement of the inflammatory status of the patients. rhamnosus GR-1 and Lactobacillus fermentum RC-14 had
Lactobacillus rhamnosus LGG given to infants during a positive impact on vaginal health [38], Lactobacillus
episodes of acute rotavirus diarrhea resulted in greater paracasei ST11 improved recovery of skin immune
increase in non-specific antibody secreting cells and homeostasis [39], and Streptococcus salivarius K12
specific anti-rotavirus antibodies in the circulation than improved oral malodour parameters [40]. Signalling of
that seen in the placebo group and resulted in shorter intestinal microorganisms to the gut–brain axis is an
duration of the diarrhea [23,24]. Furthermore, beneficial actively emerging field of research [41]. Apart from
immunomodulatory effects of specific probiotics have animal studies that showed impact of probiotics on
been observed for H. pylori-associated gastritis [25], de- anxiety, mood, and behavior, the first human trials
velopment of allergies, or reduction of allergy symptom suggest that probiotic interventions may modulate mood
scores (see [26]). and stress induced gastro-intestinal symptoms [42,43].

Immune stimulatory effects have been established in Probiotic production process and probiotic
generally healthy population as well. An improved functionality
specific immune response to a S. typhi oral vaccine has Cultivation of probiotics
been described in individuals consuming a probiotic mix In addition to providing added value to food, probiotics
containing L. johnsonii La1 and B. lactis BB12 [27] and to need to be cost effectively produced, which implies
influenza vaccine in subjects receiving L. fermentum maximizing substrate-to-biomass yield and stability
CECT5716 [28]. Furthermore, Natural Killer (NK) and during processing and shelf life.
phagocytic cell activity was increased in healthy elderly or
adults upon ingestion of B. lactis HN019 [29] or L. In the present state of knowledge, it remains difficult to
johnsonii La1 [30,31], respectively. Finally, a few studies anticipate to which extent growth conditions of probiotics
have shown a link between activated immune markers may affect their functional properties. Therefore, the
and improved resistance to infections. For example the consequences of changes in growth conditions to achieve
probiotic mix containing L. gasseri PA16/8, B. longum higher biomass yield, which may for example alter bac-
SP07/3 and B. bifidum MF 20/5 shortened the duration terial components with purported probiotic activity, may
of common cold episodes and reduced fever, while at the be overlooked. For instance, Gitton et al. [44] highlighted
same time increasing the number of leucocytes, lympho- that the global proteomic pattern differed when L. lactis
cytes (specially T-lymphocytes) and monocytes [8]. Stu- was cultivated in M17Lac broth, milk microfiltrate, or
dies such as the latter, which demonstrate a link between milk. Moreover, the time of harvesting may also influence
immune markers and improved resistance to disease will the exerted functional properties. Fayol-Messaoudi et al.

www.sciencedirect.com Current Opinion in Biotechnology 2010, 21:175–181

178 Food biotechnology

[45] demonstrated that apart from growth temperature tion and taste deterioration have to be prevented. Among
the in vitro anti-pathogen activity of the studied probio- several factors, oxygen has been identified as the key one
tics depends largely on the growth stage at which the cells that influences survival in liquid products. Strategies to
were collected. Similarly, the expression profiles of protect probiotics against oxygen were discussed else-
human mucosa (duodenal biopsies) displayed differences where [51].
in modulation of NF-kB-dependent pathways after con-
sumption of L. plantarum harvested at different growth Rehydration
phases or when heat treated [46]. A last topic related to probiotic stability and viability
assessment, which is often neglected, is the influence
Thus it would be of great advantage to identify bioactive of the rehydration step. Several studies established that
components of probiotics that can be measured under depending on the applied reconstitution conditions such
different growth conditions in order to ensure that pro- as buffer [52], pH, duration, sugar content [53], and
biotic functionality will be optimized. rehydration temperature [54], the difference in the final
cell count could vary up to 1 log cycle. These observations
Stability throughout process and storage indicate that a large proportion of the probiotic bacteria
For production of probiotics in dried form, which provides may be killed or made unculturable depending on the
a longer shelf life than liquid products, the challenge is to rehydration conditions. Hence, the conditions of the
master loss of viability due to removal of water, exposure applied enumeration methods are relevant for the
to oxygen, and eventually high temperature during dry- interpretation of the stability and viability data of pro-
ing. Further, the stability over shelf life period, which is biotics.
dictated by physical parameters of the final product
matrix and the storage conditions, has to be ensured. Furthermore, it remains to be answered to what extent
In most cases viability loss during storage is more drastic the living, the non-culturable and/or non-replicating pro-
than during processing. Improvement of stability in pow- biotics contribute to delivering a functional benefit. In the
ders can be achieved for instance by use of protective past years studies have reported that non-replicating
agents/encapsulation material, application of mild probiotics may still deliver specific health benefits
environmental sublethal stresses either during or after [55]. It is noteworthy that probably all products contain-
fermentation as recently reviewed by Muller et al. [47]. ing live probiotics will also contain a portion of dead or
damaged cells. Many of these products are down stream
Use of protective agents is a well-known strategy to processed in presence of at least part of the spent culture
increase the drying tolerance of strains. However, the medium. Hence, it cannot always be ascertained if the
potential impact on the physiology of the bacterial cells claimed functional effects of probiotic preparations are
should also be considered. For instance, Reddy et al. [48] delivered by the biomass, intracellular or extracellular cell
reported variation in in vitro cholesterol assimilation and components, and/or media derived bioactive compounds
acid tolerance upon spray drying of probiotics with differ- (Table 1).
ent types of protective agents or carriers. Finally, different
matrixes may influence the survival and functionality of For these reasons, it is presently recommended to con-
probiotics. Corcoran et al. [49] published that the presence duct clinical trials with the final formulation of the ingre-
of a metabolizable sugar could markedly enhance survival dient or product until predictive and validated functional
of L. rhamnosus LGG in simulated stomach conditions. bioassays can be applied.

Alternative strategies to address this type of questions Safety of probiotics

start to emerge. For example, accelerated evolution was The application of probiotic microorganisms in foodstuffs
applied to select a heat-shock resistant ‘spontaneous’ requires a thorough safety assessment. Several guidelines
mutant of B. longum NCC 2705 [50]. The differential are available on how to assess the safety of probiotics used
transcriptomic profile of mutant and wild type strains in food applications [56–59].
highlighted the constitutive overexpression of the classi-
cal heat-shock regulon dnaK in the mutant, which has The European Food Safety Authority (EFSA) has devel-
further proven to exhibit higher survival during down oped the QPS (Qualified Presumption of Safety)
stream processing (W. Sybesma, unpublished data). approach as a tool for the safety assessment of microor-
Importantly, the mutations introduced in the strain did ganisms used in food. This is based on a documented
not influence its functionality, as far as was demonstrated history of use and knowledge of potential pathogenic or
in a rotavirus induced diarrhea mouse model (N. Pagé, L. toxicogenic properties associated to a particular genus and
Hammarström, personal communication). species. In the US, probiotic microorganisms would be
assessed via the GRAS (Generally Recognized As Safe)
With regard to liquid probiotic application, beyond main- system. For example, B. lactis BB12, L. rhamnosus LGG,
tenance of high levels of viable cells, also post-acidifica- L. reuteri DSM 17938 strains have over recent years been

Current Opinion in Biotechnology 2010, 21:175–181 www.sciencedirect.com

 Probiotics for food Jankovic et al. 179

Table 2

Fermented yogurt drinks

Heat treated lactobacilli,

Spent culture medium,
Dried culture powders
Products/Examples

for example, see [64]

for example, see [65]

Comparison of assessment schemes for microorganisms used
in food by US FDA GRAS and EU EFSA QPS systems.

with probiotics
GRAS guidelines QPS guidelines
Applies to food additives in Applies to microorganisms
general only
Determination of GRAS status by Determination of QPS status
FDA and/or external experts by EFSA
Open list Positive list
Based on common use Based on history of use and

by cell components
adverse effects
Killed probiotic

direct contact or
Describes specific substance Describes taxonomic unit

Activity through
or microorganism (e.g. genus, species, or strain)
cell

Case-by-case assessment General assessment

Adapted from Wassenaar 2008 [66].

Yes

Yes

Yes
No

accepted in the US as GRAS for their intended use. The

main differences between the two approaches are sum-
peptides, cell wall

marized in Table 2. EU-funded research projects such as

components
Extracellular

Bacteriocins,
antimicrobial

components

ACE-ART [60] and PRO-SAFE [61] addressed the issue

Potential Probiotic Derived Bioactive Compounds

of antibiotic genes in probiotic and starter strains, and

Different concepts of probiotic production processes and related potential probiotic derived bioactive compound.

initiated an evaluation of assays that can be used to assess

Yes

Yes

Yes

Yes

biosafety of probiotics.

Data requirements to assess the safety of probiotics can

DNA, cellular
components
Intracellular

vary depending on the bacterial species of interest, the

proteins or
peptides

intended application/use and/or the target populations.

Parameters such as taxonomy and identification, pheno-
Yes

Yes

Yes
No

typic characterization, history of food use, and human

exposure, and so on, are generally considered important
[62,63]. If no history of safe use can be demonstrated,
from carbohydrates,
Fermented medium

Lactic acid derived

extensive preclinical studies, including standard 90-day

bioactive peptides
derived from milk
component

toxicity studies as defined in OECD Testing Guideline

proteins or raw

408, should also be considered. Clinical studies should

materials

include parameters to demonstrate safety in use or toler-

ability in the target population(s).
Yes

Yes

Yes
No

Rare cases of adverse effects linked to probiotic admin-

For example, frozen, spray dried, freeze dried, encapsulated.

istration have been documented in individuals having

metabolites or cell
Signalling through

serious underlying disease. Thus, special care must be

Living probiotic

activity through
components or

direct contact

taken with particularly vulnerable target population(s)

cell

such as neonates, immunocompromised subjects, or cri-

tically ill/hospitalized patients [63]. Overall the vast
Yes

Yes

amount of available data and long history of use in food-

No

No

stuffs have not indicated any safety concerns for currently

used probiotics (mainly lactobacilli and bifidobacteria) in
healthy populations.
Cultivated, killed and downstream
Fermented product as a whole

Potential mode of action/main

processed* without culture
Cultivated and down stream

Cultivated and down stream

processed with or without

Conclusion and outlook

processed* without cells

The role of functional foods is to promote the host’s

health or to restore it when it is transiently affected.
active component
Production Process

culture medium

Demonstration of the health beneficial effects of probio-

tics in generally healthy humans currently remains a
challenging task due to the lack of validated biomarkers
medium
Table 1

of health and risk factors of diseases, and the need to

undertake generally large and long-term human trials. An
*

additional challenge lies in probiotic production in a cost

www.sciencedirect.com Current Opinion in Biotechnology 2010, 21:175–181

180 Food biotechnology

effective way that will ensure probiotic viability over shelf 11. Penders J, Stobberingh EE, van den Brandt PA, Thijs C: The role
of the intestinal microbiota in the development of atopic
life. One should keep in mind that inclusion of probiotics disorders. Allergy 2007, 62:1223-1236.
in increasingly diverse food vehicles requires product and 12. Sokol H, Pigneur B, Watterlot L, Lakhdari O, Bermudez-
process modifications that may bear the risk of modulat-  Humaran LG, Gratadoux JJ, Blugeon S, Bridonneau C, Furet JP,
ing the probiotic functionality, as suggested so far mostly Corthier G et al.: Faecalibacterium prausnitzii is an anti-
inflammatory commensal bacterium identified by gut
by in vitro testing. microbiota analysis of Crohn disease patients. Proc Natl Acad
Sci USA 2008, 105:16731-16736.
Therefore research and development in the probiotic area 13. Rajilic-Stojanovic M, Heilig HG, Molenaar D, Kajander K,
should dedicate sustained efforts in the development and  Surakka A, Smidt H, de Vos WM: Development and
application of the human intestinal tract chip, a phylogenetic
validation of biomarkers of health and disease, the design microarray: analysis of universally conserved phylotypes in
of functional assays with predictive value, and the identi- the abundant microbiota of young and elderly adults. Environ
Microbiol 2009.
fication of bioactive molecules in probiotic products. This
will strongly depend on continuous efforts to identify 14. Qin J, Li R, Raes J, Arumugam M, Burgdorf KS, Manichanh C,
Nielsen T, Pons N, Levenez F, Yamada T et al.: A human gut
mechanisms of action of probiotics. With these tools in microbial gene catalogue established by metagenomic
hands it will be able to rationalize and optimize selection sequencing. Nature 2010, 464:59-65.
of probiotic candidates and downstream processing, and 15. Verberkmoes NC, Russell AL, Shah M, Godzik A, Rosenquist M,
to evaluate potential effects of the food matrixes on  Halfvarson J, Lefsrud MG, Apajalahti J, Tysk C, Hettich RL,
Jansson JK: Shotgun metaproteomics of the human distal gut
probiotic functionality. This in turns should help to better microbiota. ISME J 2009, 3:179-189.
design the human trials required for health claim sub-
16. Nicholson JK, Lindon JC: Systems biology: metabonomics.
stantiation. Nature 2008, 455:1054-1056.
17. Gloux K, Leclerc M, Iliozer H, L’Haridon R, Manichanh C,
Acknowledgement Corthier G, Nalin R, Blottiere HM, Dore J: Development of high-
We would like to thank Dr. Anne Constable for critical reviewing the safety throughput phenotyping of metagenomic clones from the
section of the manuscript. human gut microbiome for modulation of eukaryotic cell
growth. Appl Environ Microbiol 2007, 73:3734-3737.
References and recommended reading 18. Duerkop BA, Vaishnava S, Hooper LV: Immune responses to the
Papers of particular interest, published within the period of review, microbiota at the intestinal mucosal surface. Immunity 2009,
have been highlighted as: 31:368-376.
19. Gionchetti P, Rizzello F, Helwig U, Venturi A, Lammers KM,
 of special interest Brigidi P, Vitali B, Poggioli G, Miglioli M, Campieri M: Prophylaxis
 of outstanding interest of pouchitis onset with probiotic therapy: a double-blind,
placebo-controlled trial. Gastroenterology 2003, 124:1202-1209.

1. Bottazzi V: Other Fermented Dairy Products. Biotechnology. 20. Gionchetti P, Rizzello F, Morselli C, Poggioli G, Tambasco R,
Weinheim: Verlag Chemie; 1983:. pp. 315–363. Calabrese C, Brigidi P, Vitali B, Straforini G, Campieri M: High-
dose probiotics for the treatment of active pouchitis. Dis Colon
2. Metchnikoff E: The Prolongation of Life. Optimistic Studies. Rectum 2007, 50:2075-2082.
London: Butterworth-Heinemann; 1907.
21. Miele E, Pascarella F, Giannetti E, Quaglietta L, Baldassano RN,
3. Fuller R: Probiotics in man and animals. J Appl Bacteriol 1989, Staiano A: Effect of a probiotic preparation (VSL#3) on
66:365-378. induction and maintenance of remission in children with
ulcerative colitis. Am J Gastroenterol 2009, 104:437-443.
4. Dhabhar FS: Enhancing versus suppressive effects of stress on
immune function: implications for immunoprotection and 22. Pronio A, Montesani C, Butteroni C, Vecchione S, Mumolo G,
immunopathology. Neuroimmunomodulation 2009, 16:300-317. Vestri A, Vitolo D, Boirivant M: Probiotic administration in
patients with ileal pouch-anal anastomosis for ulcerative
5. Manichanh C, Rigottier-Gois L, Bonnaud E, Gloux K, Pelletier E,
colitis is associated with expansion of mucosal regulatory
Frangeul L, Nalin R, Jarrin C, Chardon P, Marteau P et al.:
cells. Inflamm Bowel Dis 2008, 14:662-668.
Reduced diversity of faecal microbiota in Crohn’s disease
revealed by a metagenomic approach. Gut 2006, 55:205-211. 23. Majamaa H, Isolauri E, Saxelin M, Vesikari T: Lactic acid bacteria
6. Wang SX, Wu WC: Effects of psychological stress on small in the treatment of acute rotavirus gastroenteritis. J Pediatr
intestinal motility and bacteria and mucosa in mice. World J Gastroenterol Nutr 1995, 20:333-338.
Gastroenterol 2005, 11:2016-2021. 24. Kaila M, Isolauri E, Soppi E, Virtanen E, Laine S, Arvilommi H:
7. Ferguson LR: Nutrigenomics approaches to functional foods. J Enhancement of the circulating antibody secreting cell
Am Diet Assoc 2009, 109:452-458. response in human diarrhea by a human Lactobacillus strain.
Pediatr Res 1992, 32:141-144.
8. de Vrese M, Winkler P, Rautenberg P, Harder T, Noah C, Laue C,
Ott S, Hampe J, Schreiber S, Heller K, Schrezenmeir J: Probiotic 25. Pantoflickova D, Corthesy-Theulaz I, Dorta G, Stolte M, Isler P,
bacteria reduced duration and severity but not the incidence Rochat F, Enslen M, Blum AL: Favourable effect of regular
of common cold episodes in a double blind, randomized, intake of fermented milk containing Lactobacillus johnsonii on
controlled trial. Vaccine 2006, 24:6670-6674. Helicobacter pylori associated gastritis. Aliment Pharmacol
Ther 2003, 18:805-813.
9. Langhendries JP, Detry J, Van HJ, Lamboray JM, Darimont J,
Mozin MJ, Secretin MC, Senterre J: Effect of a fermented infant 26. Prescott SL, Bjorksten B: Probiotics for the prevention or
formula containing viable bifidobacteria on the fecal flora treatment of allergic diseases. J Allergy Clin Immunol 2007,
composition and pH of healthy full-term infants. J Pediatr 120:255-262.
Gastroenterol Nutr 1995, 21:177-181.
27. Link-Amster H, Rochat F, Saudan KY, Mignot O, Aeschlimann JM:
10. Ley RE, Turnbaugh PJ, Klein S, Gordon JI: Microbial ecology: Modulation of a specific humoral immune response and
human gut microbes associated with obesity. Nature 2006, changes in intestinal flora mediated through fermented milk
444:1022-1023. intake. FEMS Immunol Med Microbiol 1994, 10:55-63.

Current Opinion in Biotechnology 2010, 21:175–181 www.sciencedirect.com

 Probiotics for food Jankovic et al. 181

28. Olivares M, Diaz-Ropero MP, Sierra S, Lara-Villoslada F, Fonolla J, 45. Fayol-Messaoudi D, Berger CN, Coconnier-Polter MH,
Navas M, Rodriguez JM, Xaus J: Oral intake of Lactobacillus Lievin-Le MV, Servin AL: pH-Lactic acid-, and non-lactic acid-
fermentum CECT5716 enhances the effects of influenza dependent activities of probiotic Lactobacilli against
vaccination. Nutrition 2007, 23:254-260. Salmonella enterica Serovar Typhimurium. Appl Environ
Microbiol 2005, 71:6008-6013.
29. Arunachalam K, Gill HS, Chandra RK: Enhancement of natural
immune function by dietary consumption of Bifidobacterium 46. van Baarlen P, Troost FJ, van HS, van der MC, de Vos WM, de
lactis (HN019). Eur J Clin Nutr 2000, 54:263-267.  Groot PJ, Hooiveld GJ, Brummer RJ, Kleerebezem M: Differential
NF-kappaB pathways induction by Lactobacillus plantarum in
30. Donnet-Hughes A, Rochat F, Serrant P, Aeschlimann JM, the duodenum of healthy humans correlating with immune
Schiffrin EJ: Modulation of nonspecific mechanisms of tolerance. Proc Natl Acad Sci USA 2009, 106:2371-2376.
defense by lactic acid bacteria: effective dose. J Dairy Sci 1999,
82:863-869. 47. Muller JA, Ross RP, Fitzgerald G, Stanton C: Manufacture of
 probiotic bacteria. In Prebiotics and Probiotics Science and
31. Schiffrin EJ, Brassart D, Servin AL, Rochat F, Donnet-Hughes A: Technology. Edited by Charalampopoulos D, Rastall RA. Springer;
Immune modulation of blood leukocytes in humans by lactic 2009:725-759.
acid bacteria: criteria for strain selection. Am J Clin Nutr 1997,
66:515S-520S. 48. Reddy KBPK, Madhu AN, Prapulla SG: Comparative survival and
evaluation of functional probiotic properties of spray-dried
32. Spiegel BM: The burden of IBS: looking at metrics. lactic acid bacteria. Int J Dairy Technol 2009, 62:240-248.
Curr Gastroenterol Rep 2009, 11:265-269.
49. Corcoran BM, Stanton C, Fitzgerald GF, Ross RP: Survival of
33. O’Mahony L, McCarthy J, Kelly P, Hurley G, Luo F, Chen K,  probiotic lactobacilli in acidic environments is enhanced in the
 O’Sullivan GC, Kiely B, Collins JK, Shanahan F, Quigley EM: presence of metabolizable sugars. Appl Environ Microbiol 2005,
Lactobacillus and bifidobacterium in irritable bowel 71:3060-3067.
syndrome: symptom responses and relationship to cytokine
profiles. Gastroenterology 2005, 128:541-551. 50. Berger B, Moine D, Mansourian R, Arigoni F: HspR mutations are
 naturally selected in Bifidobacterium longum when applying
34. Kajander K, Korpela R: Clinical studies on alleviating the successive heat-shock treatments. J Bacteriol 2009,
symptoms of irritable bowel syndrome. Asia Pac J Clin Nutr 192:256-263.
2006, 15:576-580.
51. Talwalkar A, Kailasapathy K: A review of oxygen toxicity in
35. De Paula JA, Carmuega E, Weill R: Effect of the ingestion of a probiotic yogurts: influence on the survival of probiotic
symbiotic yogurt on the bowel habits of women with bacteria and protective techniques. Compr Rev Food Sci Food
functional constipation. Acta Gastroenterol Latinoam 2008, Saf 2004, 3:117-124.
38:16-25.
52. Abe F, Muto M, Yaeshima T, Iwatsuki K: Effects of suspension–
36. Guyonnet D, Chassany O, Ducrotte P, Picard C, Mouret M, dilution buffers and plating media on enumeration of
Mercier CH, Matuchansky C: Effect of a fermented milk Bifidobacterium. Milchwissenschaft 2009, 64:139-142.
containing Bifidobacterium animalis DN-173 010 on the
health-related quality of life and symptoms in irritable bowel 53. Muller JA, Stanton C, Sybesma W, Fitzerald GF, Ross RP:
syndrome in adults in primary care: a multicentre, randomized, Reconstitution conditions for dried probiotic powders
double-blind, controlled trial. Aliment Pharmacol Ther 2007, represent a critical step in determining cell viability.
26:475-486. J Appl Microbiol 2010, 108:1369-1379.
37. Indrio F, Riezzo G, Raimondi F, Bisceglia M, Cavallo L, 54. Mille Y, Obert JP, Beney L, Gervais P: New drying process for
 Francavilla R: The effects of probiotics on feeding tolerance, lactic bacteria based on their dehydration behavior in liquid
bowel habits, and gastrointestinal motility in preterm medium. Biotechnol Bioeng 2004, 88:71-76.
newborns. J Pediatr 2008, 152:801-806.
55. Kataria J, Li N, Wynn JL, Neu J: Probiotic microbes: do they
38. Anukam K, Osazuwa E, Ahonkhai I, Ngwu M, Osemene G,  need to be alive to be beneficial? Nutr Rev 2009, 67:546-550.
Bruce AW, Reid G: Augmentation of antimicrobial
metronidazole therapy of bacterial vaginosis with oral 56. Food and Agriculture Organization, World Health Organization.
probiotic Lactobacillus rhamnosus GR-1 and Lactobacillus Guidelines for the evaluation of probiotics in food. 2002. Ontario,
reuteri RC-14: randomized, double-blind, placebo controlled Canada, Food and Agriculture Organization of the United Nations
trial. Microbes Infect 2006, 8:1450-1454. and World Health Organization. 11-27-2009. Ref Type: Report.
39. Peguet-Navarro J, Dezutter-Dambuyant C, Buetler T, Leclaire J, 57. Agence Française de Sécurité Sanitaire des Aliments (AFSSA).
 Smola H, Blum S, Bastien P, Breton L, Gueniche A: Alimentation infantile et modification de la flore intestinale. 2003.
Supplementation with oral probiotic bacteria protects human Ref Type: Report.
cutaneous immune homeostasis after UV exposure-double
blind, randomized, placebo controlled clinical trial. 58. Agence Française de Sécurité Sanitaire des Aliments (AFSSA).
Eur J Dermatol 2008, 18:504-511. Effets des probiotiques et prébiotiques sur la flore et l’immunité de
l’homme adulte. 2005. Ref Type: Report.
40. Burton JP, Chilcott CN, Moore CJ, Speiser G, Tagg JR: A
preliminary study of the effect of probiotic Streptococcus 59. BgVV - Bundesinstitut für gesundheitlichen Verbraucherschutz
salivarius K12 on oral malodour parameters. J Appl Microbiol und Veterinärmedizin. Final Report Working Group: Probiotic
2006, 100:754-764. Microorganisms in Food. 1999. Ref Type: Report.
41. McLean PG, Calver AR, Alpers DH, Collins SM, Shanahan F, Lee K: 60. European Commission-funded project number: CT-2003-506214.
 The emerging role of the microbial-gastrointestinal-neural Assessment and Critical Evaluation of Antibiotic Resistance
axis. Gastroenterology Insights 2009, 1:5-13. Transferability in Food Chain (ACE-ART). 2004. 11-30-0090.
Ref Type: Internet Communication.
42. Benton D, Williams C, Brown A: Impact of consuming a milk
drink containing a probiotic on mood and cognition. 61. Vankerckhoven V, Huys G, Vancanneyt M, Vael C, Klare I,
Eur J Clin Nutr 2007, 61:355-361. Romond MB, Entenza JM, Moreillon P, Wind RD, Knol J et al.:
Biosafety assessment of probiotics used for human
43. Diop L, Guillou S, Durand H: Probiotic food supplement reduces consumption: recommendations from the EU-PROSAFE
stress-induced gastrointestinal symptoms in volunteers: a project. Trends Food Sci Technol 2008, 19:102-114.
double-blind, placebo-controlled, randomized trial.
Nutr Res 2008, 28:1-5. 62. Sorokulova I: Preclinical testing in the development of
probiotics: a regulatory perspective with Bacillus strains as an
44. Gitton C, Meyrand M, Wang J, Caron C, Trubuil A, Guillot A, example. Clin Infect Dis 2008, 46(Suppl. 2):S92-S95.
Mistou MY: Proteomic signature of Lactococcus lactis
NCDO763 cultivated in milk. Appl Environ Microbiol 2005, 63. Snydman DR: The safety of probiotics. Clin Infect Dis 2008,
71:7152-7163. 1:S104-S111.

www.sciencedirect.com Current Opinion in Biotechnology 2010, 21:175–181