The Lactobacillus plantarum strain ACA-DC287 isolated from a Greek cheese demonstrates antagonistic activity in vitro and in vivo against Salmonella enterica serovar Typhimurium

Journal of Applied Microbiology ISSN 1364-5072

ORIGINAL ARTICLE

The Lactobacillus plantarum strain ACA-DC287 isolated from a Greek cheese demonstrates antagonistic activity in vitro and in vivo against Salmonella enterica serovar Typhimurium D. Fayol-Messaoudi, M.-H. Coconnier-Polter, V. Lievin-Le Moal, F. Atassi, C.N. Berger and A.L. Servin Institut National de la Sante´ et de la Recherche Me´dicale (INSERM), Unite´ 756, Faculte´ de Pharmacie, Universite´ Paris-Sud, Chaˆtenay-Malabry, France

Keywords anti-bacterial, Caco-2, cheese Lactobacillus, intestine, probiotics, Salmonella. Correspondence Alain L. Servin, Faculte´ de Pharmacie, Inserm Unite´ 756, F-92296 Chaˆtenay-Malabry, France. E-mail: [email protected]

2006 ⁄ 0992: received: 11 July 2006, revised 24 November 2006 and accepted 4 December 2006 doi:10.1111/j.1365-2672.2007.03293.x

Abstract Aims: The purpose of this study was to investigate the antibacterial activity of the Xynotyri cheese isolate Lactobacillus plantarum ACA-DC287 using a set of in vitro and in vivo assays. Methods and Results: The co-culture of L. plantarum strain ACA-DC287 and Salmonella enterica serovar Typhimurium strain SL1344 results in the killing of the pathogen. The killing activity was produced mainly by non-lactic acid molecule(s) that were present in the cell-free culture supernatant of the L. plantarum strain ACA-DC287. The culture of the L. plantarum strain ACADC287 inhibited the penetration of S. typhimurium SL1344 into cultured human enterocyte-like Caco-2 ⁄ TC7 cells. In conventional mice infected with S. typhimurium SL1344, the intake of L. plantarum strain ACA-DC287 results in a decrease in the levels of Salmonella associated with intestinal tissues or those present in the intestinal contents. In germ-free mice, the L. plantarum strain ACA-DC287 colonized the gastrointestinal tract. Conclusions: The L. plantarum strain ACA-DC287 strain exerts anti-Salmonella activity similar that of the established probiotic strains Lactobacillus rhamnosus GG, Lactobacillus casei Shirota YIT9029 and Lactobacillus johnsonii La1. Significance and Impact of the Study: The observation that a selected cheese Lactobacillus strain exerted antibacterial activity that was similar to those of probiotic Lactobacillus strains, is of interest for the use of this strain as an adjunct strain for the production of health-giving cheeses.

Introduction There is increasing evidence that the antibacterial activities of the lactobacilli involve numerous mechanisms of action, including the production of H2O2, metabolites, and antimicrobial substances, including bacteriocins and nonbacteriocin molecules (Servin 2004; Reid et al. 2003). Some probiotic Lactobacillus strains affect the adhesiveness of bacterial pathogens in a way that enables them to inhibit the bacterial invasion within cultured human epithelial cells that mimick the situation in vivo. Moreover, the antibacterial activity of selected Lactobacillus strains

has been investigated principally using two infected mouse models – the gnotobiotic or the conventional mice. Finally, a few number of probiotic strains of Lactobacillus have been reported to be effective against microbial pathogens involved in diarrhoea and urovaginal infection. The evolution of the microbiota of cheeses from various continents has been investigated showing the presence of Lactobacillus delbrueckii ssp. lactis, Lactobacillus casei, Lactobacillus paracasei ssp. paracasei, L. casei ssp. casei, Lactobacillus rhamnosus, Lactobacillus fermentum, Lactobacillus curvatus, Lactobacillus plantarum and Lactobacillus helveticus strains (Bouton et al. 1998; Morea

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et al. 1998; Xanthopoulos et al. 2000; De Vuyst et al. 2002; Antonsson et al. 2003; Manolopoulou et al. 2003; Bulut et al. 2005). It has been clearly established that the antimicrobial activity of lactobacilli is a strain-specific property, and cannot be extrapolated to other strains. The properties of Lactobacillus strains of cheese origin, including the antibacterial activity, have been poorly documented. The strain L. plantarum ACA-DC287 is a Xynotyri cheese isolate that has been selected from amongst 123 Lactobacillus strains isolated from a range of Greek food products, including raw milk, sour milk, yoghurt, Bulgarian-type yoghurt, cheese, fresh Feta curd, Feta brine, Feta cheese, Kasseri cheese, Xynotyri cheese and Galotyri cheese, on the basis of their inhibitory activity against the gram-negative pathogen Salmonella typhimurium using the well-diffusion assay (De Vuyst et al. 2004). We investigate here the antibacterial activity of this Lactobacillus strain. To do this, we have developed a set of in vitro assays, including the killing of gram-negative pathogens, activity against bacterial motility, inhibition of bacterial penetration into cultured human enterocyte-like Caco-2 ⁄ TC7 cells, and in vivo assays in mice that had already been used to investigate the activities of probiotic strains (Servin 2004). Materials and methods Bacterial strains Lactobacillus johnsonii strain La1 was from the Nestle Research Center at Vers-chez-les-Blanc (Switzerland). Lactobacillus rhamnosus strain GG was a gift from S.L. Gorbach (Tufts University, USA). Lactobacillus casei Shirota strain YIT9029 was from Yakult Honsha Co., Ltd. (Japan). The Xynotyri cheese isolate L. plantarum ACADC287 was provided by the Laboratory of Dairy Research, Department of Food Science and Technology (Agricultural University of Athens, Greece). The Lactobacillus strains were grown in 10 ml of de Man, Rogosa, Sharpe (MRS) broth (Difco Laboratories, Detroit, MI, USA) for 24 h at 37C. Lactobacillus cell-free culture supernatants (Lactobacillus CFCS) were obtained by centrifuging at 10 000 g, for 30 min at 4C. After being centrifuged, CFCS were passed through a sterile 0Æ22 lm Millex GS filter unit (Millipore, Molsheim, France). As the various Lactobacillus cultures exhibited pH values ranging from 3Æ9 to 4Æ5, a control was used in these experiments that consisted of acidified MRS at pH 4Æ5 (MRS-HCl). Moreover, as metabolites, such as lactic acid, have been shown to inhibit the growth of pathogens and even to kill them (Vandenbergh 1993), another control was used that consisted of MRS containing 60-mmol l-1 lactic acid (MRS-LA), which was equivalent to the concentration 658

found in the 24-h culture of the Lactobacillus strains used. Salmonella typhimurium SL1344 (Finlay and Falkow 1990) was from B.A.D. Stocker (Stanford University, California, USA). SL1344 was grown for 24 h at 37C in Luria broth (Difco Laboratories, Detroit, MI, USA). Lactic acid determination A commercial d- and l-lactic acid determination kit (Test-Combination d-lactic acid ⁄ l-lactic acid UV-method, Boehringer Mannheim GmbH, Germany) was used to determine the concentration of lactic acid in the Lactobacillus CFCS. Determination of the killing activity An exponentially growing culture of S. typhimurium SL1344 was centrifuged at 5.500 g for 5 min at 4C. The culture medium was discarded, and the bacteria were washed once with phosphate-buffered saline (PBS), and then resuspended in Luria broth. Bacterial cells were counted in a Salumbini chamber, and then adjusted to the desired concentration before experiments. The total killing activity of the Lactobacillus cultures or CFCS was determined in the presence of Luria broth as previously described (Coconnier et al. 1997). The colony-count assays were performed by incubating 500 ll of S. typhimurium SL1344 at 2 · 108 colony forming units per ml (CFU ml-1), with 500 ll of Lactobacillus culture or CFCS, MRS, MRS-HCl (pH 4Æ5), or MRS-LA (60 mmol l-1, pH 4Æ5) for 4 h at 37C (pH of incubation medium 45 ± 0Æ2). The aliquots were removed, serially diluted, and plated on trypticase soy agar (TSA) (Invitrogen, Cergy, France) to determine the bacterial colony count. In order to investigate the killing activity of the nonlactic acid molecule(s) present in the Lactobacillus CFCS, we used a recently described, in vitro method that distinguishes between the lactic acid- and non-lactic aciddependent anti-Salmonella activities of Lactobacillus strains (Fayol-Messaoudi et al. 2005). The S. typhimurium SL1344 bacteria were resuspended in Dulbecco’s modified Eagle’s minimum essential medium (DMEM) (Invitrogen), and bacteria were counted in a Salumbini chamber, and then adjusted to the desired concentration. A colonycount assay was performed by incubating 500 ll of S. typhimurium SL1344 at 2 · 108 CFU ml-1, with 500 ll of Lactobacillus CFCS, or MRS, MRS-HCl (pH 4Æ5), or MRS-LA (60 mmol l-1, pH 4Æ5) at 37C for 4 h (pH of the incubation medium 4Æ8 ± 0Æ2). The aliquots were removed, serially diluted, and plated on TSA to determine the bacterial colony count.

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Penetration of Salmonella into cultured human intestinal Caco-2 ⁄ TC7 cells The TC7 clone (Caco-2 ⁄ TC7) (Chantret et al. 1994), established from the parental human enterocyte-like Caco-2 cell line (Pinto et al. 1983) was used. Cells were routinely grown in DMEM (25-mmol l-1 glucose) (Invitrogen), supplemented with 15% heat-inactivated (30 min, 56C) fetal calf serum (FCS) (Invitrogen) and 1% nonessential amino acids. For maintenance purposes, the cells were passaged weekly using 0Æ02% trypsin in Ca2+Mg2+-free PBS containing 3-mmol l-1 EDTA. The experiments and cell maintenance were carried out at 37C in an atmosphere of 10% CO2–90% air. The culture medium was changed daily. For assays of S. typhimurium SL1344 infection, fully differentiated cells were used at postconfluence after 15 days in culture (Coconnier et al. 1997, 2000). The inhibition of the internalization of S. typhimurium SL1344 by Lactobacillus cultures was determined as previously described (Bernet-Camard et al. 1997). An exponentially growing culture of S. typhimurium SL1344 bacteria in DMEM (500 ll, 5 · 107 CFU per well) was incubated with the Caco-2 ⁄ TC7 cells (1 h at 37C in an atmosphere of 10% CO2–90% air) in the presence of Lactobacillus cultures (500 ll of 24-h cultures), or of MRS, MRS-HCl (pH 4Æ5), or MRS-LA (60 mmol l-1, pH 4Æ5). The plates were then washed three times with sterile PBS, and the number of viable intracellular Salmonella was determined by a colony-count assay as described earlier. After exposure to the Lactobacillus culture, the cell integrity was determined by measuring the lactate dehydrogenase (LDH) activity in the incubation medium (Enzyline LDH kit, Biome´rieux, Dardilly, France), according to the manufacturer’s instructions. The integrity of the cells incubated with Lactobacillus CFCS was the same as that of the cells incubated with DMEM alone (not shown). Anti-Salmonella activity in conventional mice The anti-Salmonella activity was determined in conventional mice which had a gastrointestinal (GI) microbiota, as previously reported (Bernet et al. 1994; Coconnier et al. 1997). Conventional C3H ⁄ He ⁄ Oujco mice (Charles River, Lyon, France), free from specific and opportunistic pathogens, such as Proteus (SOPF), and 6 or 7 weeks in age, were used. They were housed and fed in accordance with the relevant national legislation. The mice were infected with S. typhimurium SL1344 by intragastric force-feeding once (1 · 108 CFU per mouse). On two successive days, the infected mice received the same dose of a 24-h Lactobacillus culture by intragastric force-feed-

Strain ACA-DC287 inhibits Salmonella

ing (0Æ2 ml, 109 CFU ml-1). The control infected mice were given MRS. The mice were then killed by cervical dislocation. The following GI tract segments were removed: stomach, small intestine (divided into three segments corresponding approximately to the duodenum, jejunum and ileum), caecum and colon. After removing the contents, the intestinal wall was gently washed with eight successive 10-ml sterile aliquots of PBS, and drained. The tissues were weighed and homogenized for 2 min with 1 ml of PBS using an Ultraturrax. All the content samples and organs were serially diluted and plated on Shigella and Salmonella agar (Difco, Paris, France), to estimate the number of viable S. typhimurium SL1344. Black colonies of S. typhimurium were easily distinguishable. Results are reported as log 10 CFU ± SD per g of tissue or per g of content. Colonization of gut by Lactobacillus strains in germ-free mice In order to examine the colonization properties of Lactobacillus strains in a germ-free mouse model, germfree adult female C3H ⁄ He ⁄ Oujco mice, 7-8 weeks of age (Iffa Credo, L’Arbresle 69, France) were used. They were housed and fed in accordance with the relevant national legislation. The germ-free mice were reared in Trexler type isolators fitted with a rapid transfer system (La Calhe`ne, Ve´lizy Villacoublay, France). All the mice had ad libitum access to commercial chow RO3 (UAR, Villemoisson ⁄ Orge, France), which had been irradiated at 40 kGy, and to autoclaved demineralized water. The mice were checked to confirm the absence of bacterial contamination by cultures of fresh faeces under aerobic and anaerobic conditions. The mice were deprived of water from the day before exposure to infection (six mice per group). They were given orally a Lactobacillus culture in their drinking water (10-fold dilution of a fresh, 24-h culture in bottled water, i.e. approximately 3 · 108 CFU per mouse). In all cases, a high population of lactobacilli (109 CFU g-1 of fresh faeces) was detected 18 h after infection. The mice were killed by cervical dislocation and the segments of the GI tract were treated as reported earlier. All the content samples and tissues were serially diluted and plated on MRS agar, to estimate the number of viable lactobacilli. The plates were incubated at 37C for 48 h. The results are reported as log 10 CFU ± SD per g of intestinal wall or per g of content. Statistics The data are expressed as the mean ± standard deviation (SD). The statistical significance was assessed using

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Table 1 Effect of Lactobacillus cultures on the viability of Salmonella typhimurium SL1344 S. typhimurium (log CFU ml-1) Control Nonacidified MRS MRS-HCl (pH 4Æ5) MRS-LA (60 mmol l-1, pH 4Æ5) Lactobacillus rhamnosus GG Lactobacillus casei Shirota YIT9029 Lactobacillus johnsonii La1 Lactobacillus plantarum ACA-DC287

8Æ77 8Æ52 8Æ05 4Æ23 2Æ00 2Æ11 2Æ18 2Æ13

± ± ± ± ± ± ± ±

0Æ02 0Æ10 0Æ08 0Æ84* 0Æ05* 0Æ14* 0Æ21* 0Æ16*

*P < 0Æ05 versus control. The inoculum of S. typhimurium SL1344 contained 108 CFU ml-1. Viable Salmonella were counted after 4 h of co-culture with the Lactobacillus strains in the presence of Luria broth. Each value shown is the mean ± SD from three experiments. MRS, de Man, Rogosa, Sharpe.

660

*

* *

* *

ACA-DC287

*

YIT9029

*

La1

*

GG

MRS-LA

The killing activities against S. typhimurium SL1344 of the L. plantarum strain ACA-DC287 was determined. As controls, the probiotic strains L. rhamnosus GG, L. casei Shirota YIT9029 and L. johnsonii La1 were used. As shown in Table 1, the unacified MRS and acidified MRS (MRS-HCl), used as controls, displayed no more killing activity than untreated S. typhimurium SL1344 after 4 h of contact in the presence of Luria broth. The MRS containing 60-mmol l-1 lactic acid (MRS-LA), which was equivalent to the concentration present in the Lactobacillus cultures (L. rhamnosus GG: 63 ± 8; L. casei Shirota YIT9029: 64 ± 12; L. johnsonii La1: 61 ± 16; L. plantarum ACA-DC287: 68 ± 19 mmol l-1), reduced the viability of S. typhimurium SL1334 by 4Æ5 logs in the presence of Luria broth. The culture of the L. plantarum strain ACADC287 displayed killing activity, lowering the level of viable bacteria by 6Æ7 logs relative to the control S. typhimurium SL1344 in the presence of Luria broth. The cultures of L. rhamnosus GG, L. casei Shirota YIT9029 and L. johnsonii La1 strains all killed S. typhimurium SL1344 to a similar extent, reducing the level of viable bacteria by 6Æ7 logs in the presence of Luria broth. As previously reported (Fayol-Messaoudi et al. 2005), the killing activity of CFCS was determined in the presence of Luria broth (total killing activity) and in the pres-

*

MRS-HCI

Killing activity

MRS

Results

2 1 0 –1 –2 –3 –4 –5 –6 –7 Control

student’s t test. The differences were considered significant at a P value of 99Æ9% (>3 logs of decrease in viability) of a test micro-organism after being incubated with it for a fixed length of time under controlled conditions (N.C.C.L.S. 1999). There is increasing evidence that the antibacterial activities of lactic acid bacteria involve numerous mechanisms of action, including the production of H2O2, of metabolites, such as lactic acid and of antimicrobial substances, including bacteriocins and nonbacteriocin molecules (Servin 2004). We found that the killing activity of the L. plantarum strain ACA-DC287 resulted from the production of non-lactic acid antibacterial substance(s) present in its CFCS, as has previously been reported for the probiotic strains L. rhamnosus GG (Silva et al. 1987; Hudault et al. 1997; Fayol-Messaoudi et al. 2005), L. johnsonii La1 (BernetCamard et al. 1997; Fayol-Messaoudi et al. 2005;), and L. acidophilus LB (Coconnier et al. 1997, 2000; Coconnier-Polter et al. 2005). Probiotic Lactobacillus strains, including L. acidophilus LB (Chauviere et al. 1992; Coconnier et al. 1993a,b), L. rhamnosus GG (Hudault et al. 1997; Lee et al. 2000, 2003; Lee and Puong 2002), L. casei Shirota YIT9029 (Lee et al. 2000, 2003; Lee and Puong 2002), L. acidophilus 662

*

* 4.0

Figure 3 Distribution of Salmonella typhimurium SL1344 in the stomach and intestine tissues, and in the stomach and intestinal contents of infected conventional mice treated with the cheese Lactobacillus plantarum ACADC287 or probiotic Lactobacillus casei Shirota YIT9029 strains. Experimental conditions are described in Material and Methods. 1, stomach; 2, duodenum; 3, jejunum; 4, ileum; 5, caecum; 6, colon. Each point represents the mean ± SD of six mice per group. *Significant differences between the numbers of viable S. typhimurium SL1344 under the untreated (j) and treated conditions (u) (P < 0Æ05).

HN017 (Gopal et al. 2001), L. rhamnosus DR20 (Gopal et al. 2001), L. gasseri K7 (Bogovic-Matijasic et al. 2006) and L. johnsonii La1 (Bernet-Camard et al. 1997) were able to inhibit the adhesion to and ⁄ or the cell entry into cultured human intestinal cells of enterovirulent pathogens. Here, we observed that in competition condition, the culture of the L. plantarum strain ACA-DC287 was able to inhibit the penetration of S. typhimurium SL1344 into cultured human intestinal Caco-2 ⁄ TC7 cells as effectively as the probiotic strains L. rhamnosus GG, L. casei Shirota YIT9029 and L. johnsonii La1. As recently pointed out (Servin 2004; Reid and Burton 2002), it is difficult to extrapolate from in vitro effects to the GI situation in vivo. Indeed, it is recognized that the resident microbiota and mucus, and the peristaltic flow that continuously washes over the GI epithelium, could considerably modify or even abolish the effects of exogenous probiotic lactic acid bacteria. However, L. casei Shirota strain YIT9029 has been found active on oral infection with the enteric pathogen List. monocytogenes in conventional Wistar rats (de Waard et al. 2002). The oral administration of probiotic L. johnsonii La1 or L. acidophilus LB decreases the level of viable S. typhimurium in the faeces of infected conventional mice (Bernet-Camard et al. 1997; Coconnier et al. 1997). To demonstrate that

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Strain ACA-DC287 inhibits Salmonella

that the L. plantarum strain ACA-DC287 has been selected from amongst 123 Lactobacillus strains isolated from a range of Greek food products. Probiotic strains are defined as live micro-organisms which, when consumed in appropriate amounts in food, confer a health benefit on the host (FAO ⁄ WHO 2001). The observation that a selected cheese Lactobacillus strain exerted antibacterial activity, which was similar to that of probiotic Lactobacillus strains, is of interest for the use of this strain as an adjunct strain for the production of health-giving cheeses.

Lactobacillus (log CFU g–1)

9.0 YIT9029 8.0 7.0 6.0 5.0 4.0 3.0

Acknowledgements 2.0 1

2

3

4

5

6

3

4

5

6

Lactobacillus (log CFU g–1)

9.0 ACA-DC287 8.0 7.0 6.0 5.0 4.0 3.0 2.0 1

2

Figure 4 Distribution of lactobacilli in the stomach and intestine tissues, and in the stomach and intestinal contents of germ-free mice inoculated with the cheese Lactobacillus plantarum ACA-DC287 or probiotic Lactobacillus casei Shirota YIT9029 strains. Experimental conditions are described in Material and Methods. 1, stomach; 2, duodenum; 3, jejunum; 4, ileum; 5, caecum; 6, colon. Each point represents the mean ± SD of six mice per group.

the antagonistic activity of L. plantarum strain ACADC287 against S. typhimurium SL1344 observed in vitro also occur in vivo, we have used S. typhimurium-infected conventional mice. We found that the L. plantarum strain ACA-DC287 did reduce the levels of viable Salmonella associated with the intestinal epithelium and present in the intestinal contents in infected mice. The antagonistic effect correlates with the presence of L. plantarum isolate ACA-DC287 within the GI tract. In conclusion, the cheese L. plantarum strain ACADC287 exerts antibacterial activities in vitro and in vivo, which are similar to those previously reported for selected probiotic strains of human origin. It has clearly been established that the anti-microbial activity of probiotic Lactobacillus strains of human origin is a strain-specific property, and cannot be extrapolated to other strains (Servin 2004; FAO ⁄ WHO 2001). It is interesting to note

Part of this study was been carried out with financial support from the Commission of the European Communities, specific RTD programme ‘Quality of Life and Management of Living Resources’, QLK1-2001-01179 ‘PROPATH’ – Molecular analysis and mechanistic elucidation of the functionality of probiotics and prebiotics in the inhibition of pathogenic micro-organisms to combat GI disorders and to improve human health. It does not necessarily reflect its views and in no way anticipates the Commission’s future policy in this area. D. FayolMessaoudi and C.N. Berger were funded during their PhD studies by the QLK1-2001-01179 PROPATH Program (Commission of the European Communities). We are very grateful to E. Tsakalidou (Agricultural University of Athens, Greece) for his generous gift of the L. plantarum strain ACA-DC287. We are grateful to L. De Vuyst and E. Makras (IMDO, Department of Applied Biological Sciences, Vrije Universiteit Brussels, Belgium), A. Mentis and D. Sgouras (Hellenic Pasteur Institute, Laboratory of Bacteriology, Athens, Greece) and I. Nes (Laboratory of Microbial Gene Technology, Agricultural University of Norway, Aas, Norway) for stimulating discussion. References Antonsson, M., Molin, G. and Ardo, Y. (2003) Lactobacillus strains isolated from Danbo cheese as adjunct cultures in a cheese model system. Int J Food Microbiol 85, 159–169. Atanassova, M., Choiset, Y., Dalgalarrondo, M., Chobert, J.M., Dousset, X., Ivanova, I. and Haertle, T. (2003) Isolation and partial biochemical characterization of a proteinaceous anti-bacteria and anti-yeast compound produced by Lactobacillus paracasei subsp. paracasei strain M3. Int J Food Microbiol 87, 63–73. Bernet, M.F., Brassart, D., Neeser, J.R. and Servin, A.L. (1994) Lactobacillus acidophilus LA 1 binds to cultured human intestinal cell lines and inhibits cell attachment and cell invasion by enterovirulent bacteria. Gut 35, 483–489.

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Bernet-Camard, M.F., Lievin, V., Brassart, D., Neeser, J.R., Servin, A.L. and Hudault, S. (1997) The human Lactobacillus acidophilus strain LA1 secretes a nonbacteriocin antibacterial substance(s) active in vitro and in vivo. Appl Environ Microbiol 63, 2747–2753. Bogovic-Matijasic, B., Narat, M., Zoric Peternel, M. and Rogelj, I. (2006) Ability of Lactobacillus gasseri K 7 to inhibit Escherichia coli adhesion in vitro on Caco-2 cells and ex vivo on pigs’ jejunal tissue. Int J Food Microbiol 107, 92–96. Bouton, Y., Guyot, P. and Grappin, R. (1998) Preliminary characterization of microflora of Comte cheese. J Appl Microbiol 85, 123–131. Bulut, C., Gunes, H., Okuklu, B., Harsa, S., Kilic, S., Coban, H.S. and Yenidunya, A.F. (2005) Homofermentative lactic acid bacteria of a traditional cheese, Comlek peyniri from Cappadocia region. J Dairy Res 72, 19–24. Caridi, A. (2002) Selection of Escherichia coli-inhibiting strains of Lactobacillus paracasei subsp. paracasei. J Ind Microbiol Biotechnol 29, 303–308. Chantret, I., Rodolosse, A., Barbat, A., Dussaulx, E., Brot-Laroche, E., Zweibaum, A. and Rousset, M. (1994) Differential expression of sucrase-isomaltase in clones isolated from early and late passages of the cell line Caco-2: evidence for glucose-dependent negative regulation. J Cell Sci 107, 213–225. Chauviere, G., Coconnier, M.H., Kerneis, S., DarfeuilleMichaud, A., Joly, B. and Servin, A.L. (1992) Competitive exclusion of diarrheagenic Escherichia coli (ETEC) from human enterocyte-like Caco-2 cells by heat-killed Lactobacillus. FEMS Microbiol Lett 70, 213–217. Coconnier, M.H., Bernet, M.F., Chauviere, G. and 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 Diarrhoeal Dis Res 11, 235–242. Coconnier, M.H., Bernet, M.F., Kerneis, S., Chauviere, G., Fourniat, J. and 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, 299–305. Coconnier, M.H., Lievin, V., Lorrot, M. and Servin, A.L. (2000) Antagonistic activity of Lactobacillus acidophilus LB against intracellular Salmonella enterica serovar Typhimurium infecting human enterocyte-like Caco-2 ⁄ TC-7 Cells. Appl Environ Microbiol 66, 1152–1157. Coconnier, M.H., Lievin, V., Bernet-Camard, M.F., Hudault, S. and Servin, A.L. (1997) Antibacterial effect of the adhering human Lactobacillus acidophilus strain LB. Antimicrob Agents Chemother 41, 1046–1052. Coconnier-Polter, M.H., Lievin-Le Moal, V. and Servin, A.L. (2005) A Lactobacillus acidophilus strain of human gastrointestinal microbiota origin elicits killing of enterovirulent Salmonella enterica Serovar Typhimurium by triggering

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lethal bacterial membrane damage. Appl Environ Microbiol 71, 6115–6120. De Vuyst, L., Schrijvers, V., Paramithiotis, S., Hoste, B., Vancanneyt, M., Swings, J., et al. (2002) The biodiversity of lactic acid bacteria in Greek traditional wheat sourdoughs is reflected in both composition and metabolite formation. Appl Environ Microbiol 68, 6059–6069. De Vuyst, L., Makras, L., Avonts, L., Holo, H., Quing, Y., Servin, A., et al. (2004) Antimicrobial potential of probiotic or potentially probiotic lactic acid bacteria, the first results of the international european research project PROPATH of the PROEUHEALTH cluster. Microbial Ecol Health and Dis 16, 125–130. de Waard, R., Garssen, J., Bokken, G.C. and Vos, J.G. (2002) Antagonistic activity of Lactobacillus casei strain Shirota against gastrointestinal Listeria monocytogenes infection in rats. Int J Food Microbiol 73, 93–100. FAO ⁄ WHO (2001) Joint FAO ⁄ WHO Expert Consultation on Evaluation of Health and Nutritional Properties of Probiotics in Food Including Powder Milk with Live Lactic Acid Bacteria. Cordoba, Argentina 1 to 4 October 2001. http:// www.who.int/foodsafety/publications/fs_management/en/ probiotics.pdf. Fayol-Messaoudi, D., Berger, C.N., Coconnier-Polter, M.-H., Lievin-Le Moal, V. and Servin, A.L. (2005) pH-, lactic acid- and non-lactic acid-dependent activities against Salmonella enterica serovar Typhimurium by probiotic strains of Lactobacillus. Appl Environ Microbiol 71, 6008–6013. Finlay, B.B. and Falkow, S. (1990) Salmonella interactions with polarized human intestinal Caco-2 epithelial cells. J Infect Dis 162, 1096–1106. Gopal, P.K., Prasad, J., Smart, J. and Gill, H.S. (2001) In vitro adherence properties of Lactobacillus rhamnosus DR20 and Bifidobacterium lactis DR10 strains and their antagonistic activity against an enterotoxigenic Escherichia coli. Int J Food Microbiol 67, 207–216. Hudault, S., Lievin, V., Bernet-Camard, M.F. and Servin, A.L. (1997) Antagonistic activity exerted in vitro and in vivo by Lactobacillus casei (strain GG) against Salmonella typhimurium C5 infection. Appl Environ Microbiol 63, 513–518. Lee, Y.K. and Puong, K.Y. (2002) Competition for adhesion between probiotics and human gastrointestinal pathogens in the presence of carbohydrate. Br J Nutr 88, S101–108. Lee, Y.K., Puong, K.Y., Ouwehand, A.C. and Salminen, S. (2003) Displacement of bacterial pathogens from mucus and Caco-2 cell surface by lactobacilli. J Med Microbiol 52, 925–930. Lee, Y.K., Lim, C.Y., Teng, W.L., Ouwehand, A.C., Tuomola, E.M. and Salminen, S. (2000) Quantitative approach in the study of adhesion of lactic acid bacteria to intestinal cells and their competition with enterobacteria. Appl Environ Microbiol 66, 3692–3697. Manolopoulou, E., Sarantinopoulos, P., Zoidou, E., Aktypis, A., Moschopoulou, E., Kandarakis, I.G. and Anifantakis, E.M. (2003) Evolution of microbial populations during

ª 2007 The Authors Journal compilation ª 2007 The Society for Applied Microbiology, Journal of Applied Microbiology 103 (2007) 657–665

D. Fayol-Messaoudi et al.

traditional Feta cheese manufacture and ripening. Int J Food Microbiol 82, 153–161. Martin, R., Olivares, M., Marin, M.L., Xaus, J., Fernandez, L. and Rodriguez, J.M. (2005) Characterization of a reuterinproducing Lactobacillus coryniformis strain isolated from a goat’s milk cheese. Int J Food Microbiol 104, 267– 277. Miteva, V., Ivanova, I., Budakov, I., Pantev, A., Stefanova, T., Danova, S., et al. (1998) Detection and characterization of a novel antibacterial substance produced by a Lactobacillus delbrueckii strain 1043. J Appl Microbiol 85, 603– 614. Morea, M., Baruzzi, F., Cappa, F. and Cocconcelli, P.S. (1998) Molecular characterization of the Lactobacillus community in traditional processing of Mozzarella cheese. Int J Food Microbiol 43, 53–60. N.C.C.L.S. (1999) Methods for Determining Bactericidal Activity of Antimicrobial Agents: Approved Guideline. Wayne, PA: National Committee for Clinical Laboratory Standards, Wayne, PA, pp. 1–29. Pinto, M., Robine-Leon, S., Appay, M.D., Kedinger, M., Triadou, N., Dussaulx, E., et al. (1983) Enterocyte-like dif-

Strain ACA-DC287 inhibits Salmonella

ferentiation and polarization of the human colon carcinoma cell line Caco-2 in culture. Biol Cell 47, 323–330. Reid, G. and Burton, J. (2002) Use of Lactobacillus to prevent infection by pathogenic bacteria. Microbes Infect 4, 319– 324. Reid, G., Jass, J., Sebulsky, M.T. and McCormick, J.K. (2003) Potential uses of probiotics in clinical practice. Clin Microbiol Rev 16, 658–672. Servin, A.L. (2004) Antagonistic activities of lactobacilli and bifidobacteria against microbial pathogens. FEMS Microbiol Rev 28, 405–440. Silva, M., Jacobus, N., Deneke, C. and Gorbach, S.L. (1987) Antimicrobial substance from a human Lactobacillus strain. Antimicrob Agents Chemother 31, 1231–1233. Vandenbergh, P.A. (1993) Lactic acid bacteria, their metabolic products and interference with microbial growth. FEMS Microbiol Rev 12, 221–238. Xanthopoulos, V., Hatzikamari, M., Adamidis, T., Tsakalidou, E., Tzanetakis, N. and Litopoulou-Tzanetaki, E. (2000) Heterogeneity of Lactobacillus plantarum isolates from feta cheese throughout ripening. J Appl Microbiol 88, 1056– 1064.

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