Journal of Applied Microbiology ISSN 1364-5072

ORIGINAL ARTICLE

Effects of a synbiotic milk product on human intestinal ecosystem

M.C. Casiraghi1, E. Canzi2, R. Zanchi2, E. Donati3 and L. Villa3
1 Department of Food Science and Microbiology, Division of Human Nutrition, University of Milan, Milan, Italy 2 Division of Microbiology, Department of Food Science and Microbiology, University of Milan, Milan, Italy 3 Centrale del Latte di Milano, Granarolo Group, Milan, Italy

Keywords bidobacteria, faecal bile acids, inulin, lactic acid bacteria, synbiotic. Correspondence M. Cristina Casiraghi, Department of Food Science and Microbiology, Division of Human Nutrition, University of Milan, Via Celoria n.2, 20133 Milano, Italy. E-mail: maria.casiraghi@unimi.it

Abstract Aims: To investigate the effect of prolonged consumption of a synbiotic milk (Synbiotic) containing Lactobacillus acidophilus (strain 74-2, 107 CFU ml)1), Bidobacterium lactis (strain 420, 107 CFU ml)1) and 2% inulin on colonic ecosystem in healthy humans. Methods and Results: A group of 26 healthy subjects, aged 2247 years, participated in a 6-week placebo-controlled dietary intervention study. After a 2week baseline period, in which all volunteers consumed 500 ml day)1 of 2% skimmed milk (Placebo), the study was designed as a randomized, doubleblind, two-armed parallel study in which 4-week consumption of 500 ml day portions of Synbiotic or Placebo were compared. Faecal microbial counts, pH, l-lactic acid and bile acid concentrations were assessed before and after the intervention. Synbiotic consumption signicantly decreased faecal dry weight (P < 001) and l-lactic acid (P < 005) concentration, while signicantly increased faecal bidobacteria (P < 005) and lactobacilli (P < 001) counts. Conclusion: The tested synbiotic milk showed its synbiotic nature by enhancing the growth of bidobacteria and lactobacilli. Signicance and Impact of the Study: Scientic support to functional effect of a synbiotic milk.

2006/0951: received 5 July 2006, revised 5 September 2006 and accepted 6 November 2006
doi:10.1111/j.1365-2672.2006.03273.x

Introduction The awareness that food has benets over and beyond the basic nutrients that it provides has recently emerged in nutrition, and the concept of functional food i.e. foods and food ingredients that can enhance health was introduced (Diplock et al. 1999). Among functional foods those which exert a favourable effect on the colonic ecosystem have received widespread research worldwide; they include probiotics, prebiotics and synbiotics. Probiotics were dened as a live microbial food supplement which benecially affects the host by improving its intestinal microbial balance (Fuller 1989). Probiotics, most notably lactobacilli and bidobacteria, are marketed as capsules, powders, enriched yoghurts, yoghurt-like products and milks. Possible health effects of such products include improved bioavailability of nutrients, allevi-

ation of the symptoms of lactose intolerance, immune system stimulation, cholesterol lowering, and colon cancer prevention (Goldin 1998; Naidu et al. 1999). Prebiotics were dened by Gibson and Roberfroid (1995) as a nondigestible food ingredient that benecially affects the host by selectively stimulating the growth and/ or the activity of one or a limited number of bacteria in the colon. Among the various nondigestible oligosaccharides which could have this potential, inulin and oligofructose are the most studied. At the present there is convincing evidence that such substrates affect the composition and the metabolic activity of the intestinal microbiota, stimulate bowel habits and mineral absorption (Gibson et al. 2004); in addition, there are promising observations in animal models concerning chemo-prevention and retardation of tumour growth as a consequence of inulin administration (Van Loo et al. 1999).
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When combining both a probiotic and a prebiotic in a single food product, the expected benets are, as stated in the synbiotic denition, an improved survival of the probiotic bacteria during the passage through the upper intestinal tract and a more efcient implantation of probiotics in the large bowel, together with a stimulating effect of the prebiotic on the growth and/or the activities of both the exogenous (probiotic) and endogenous bacteria (e.g. bidobacteria). In accordance with Roberfroid (1998), whereas the concept of prebiotic has become very popular since its introduction in 1995, the concept of synbiotic has, up to now, not widely applied to the development of functional foods. This concept thus remains open for validation and further research is needed. The aim of this study has been to investigate, in healthy humans, the inuence of a synbiotic milk product consumption on intestinal ecosystem by estimating faecal microbiota composition. Furthermore, because a synbiotic-mediated modulation of intestinal microora could inuence host metabolic pathways, some faecal biochemical marker (bile acids, pH and l-lactate) have been also investigated. Materials and methods Synbiotic and placebo The experimental product (Synbiotic) was prepared on a pilot plan by Centrale del latte di Milano Granarolo Group, Milan, Italy, using pasteurized skimmed (2% fat) fresh milk added with 2% of inulin (mean degree of polymerization 10, Raftiline; Orafti, Tienen, Belgium), Lactobacillus acidophilus (strain 74-2, 107 CFU ml)1) and Bidobacterium lactis (strain 420, 107 CFU ml)1). The probiotic strains were from Wiesby GmbH & Co., KG, Vimercate, Italia. Skimmed milk (2% fat) without the addition of prebiotic and probiotics was used as Placebo in the trial. Inulin content and probiotic counts in Synbiotic samples, from three different production cycles, were assessed just after production and at the end of its shelf-life (5 days). Synbiotics inulin content, assessed by HPLC
Time 0 day (T0 day): Foods record Intestinal habits Faecal samples Time 14 days (T14 days): Foods record Intestinal habits Faecal samples Placebo n = 13

(Prosky and Hoebregs 1999), was 184 151 mg ml)1 (mean SD) at production and 185 149 mg ml)1 after 5 days of conservation at 4C. Viable counts of bidobacteria and lactobacilli in Synbiotic were evaluated, in duplicate, on Rogosa agar (Oxoid, Basingstoke, UK) and trypticasephytoneyeast extract agar (Scardovi 1986) respectively, after 3 days of anaerobic incubation in GasPak (bioMerieux, Marcy lEtoile, France) at 37C. At production Synbiotic contained 692 004 log CFU ml)1 and 698 011 log CFU ml)1 of L. acidophilus and B. lactis, respectively; probiotics counts did not signicantly change at the end of shelf-life, being 671 001 log CFU ml)1 for L. acidophilus and 651 037 log CFU ml)1 for B. lactis. After production the Synbiotic was kept at 4C and administered fresh to volunteers (within 4 days). Subjects A total of 26 healthy volunteers (14 females and 12 males) participated in the study, after providing written consent. Inclusion criteria were: milk consumers, normal dietary habits, excluding vegetarianism, therapeutic diets and every kind of food supplements and other specic probiotics and prebiotics; no drug therapy, no pathologies or surgery of gastrointestinal tract, no laxative of any class, no use of antibiotics within the previous six months; low to normal physical exercise (<60 min exercise three times per week). During the trial period volunteers followed their usual diet without other dietary restrictions. The study was approved by the Ethics Committee of the University of Milan. Experimental design The trial lasted for 6 weeks and was divided into two consecutive phases: a baseline period of 2 weeks and a treatment period of 4 weeks. During the baseline period (T0d and T14d) all volunteers consumed 500 ml day)1 of Placebo; the treatment period was designed as a randomized, double-blind, two-armed parallel study in which Synbiotic was compared with Placebo for a consecutive period of 4 week (T42d) (Fig. 1). Age and body mass index values of volunteers were homogeneous among
Time 42 days (T4 42 days): Foods record Intestinal habits Faecal samples

Placebo (Baseline) n = 26

( Treatment ) Synbiotic n = 13
Figure 1 Experimental design.

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Table 1 Characteristics of subjects involved in the study

Placebo Males (n) Females (n) Age (years) Body mass index (kg m)2) 6 7 305 19 (2241) 217 10 (160294) Synbiotic 6 7 354 28 (2247) 232 31 (187284)

Values are presented as mean SE; range in parenthesis.

and on Oxoid Rogosa agar. The counts of lactobacilli were assessed on two different selective media because preliminary tests showed the inability of the L. acidophilus, present in the Synbiotic, to grow on Lamvab medium in the presence of vancomycin which, however, does not inhibit most intestinal lactobacilli (Hartemink et al. 1997). Micro-organisms were classied according to V.P.I. Anaerobe Laboratory Manual (Holdeman et al. 1977) and Bergeys Manual Determinative Bacteriology (Holt et al. 1994) as previously described (Canzi et al. 2002). Faecal biochemical analyses After homogenization, pH was measured directly on fresh faecal samples with a pH electrode (mod. PH M83 Autocal; Radiometer, Copenhagen, Denmark). Faecal l-lactic acid concentration was measured by means of a dedicated analyser (model 2300; Yellow Spring Inst., Yellow Spring, OH, USA), after dilution of faecal samples with NaCl 09% solution (1/5, w/v). A sample of about 30 g of faeces was freeze-dried, weighed and 200 mg of dry sample was analysed for free bile acids concentration after ethanol extraction, according to Korpela et al. (1986), using cholanic acid (5-bcholanic acid: C7628 ursocholanic acid; Sigma, Milan, Italy) as internal standard. Briey, faecal ethanol extract was applied on a DEAE-Sephadex A-25, 5 mm 5 cm column (Pharmacia, Upsala, Sweeden); after washing with 7 ml of chloroform/70% methanol solution (15 : 10, v/v), free bile acids were eluted with 5 ml of 02 mol l)1 acetic acid in 70% methanol. This fraction was evaporated to dryness under N2 ow and bile acids were assessed by GLC, after trimethylsilylester derivatization. Analysis was performed using a Varian model 3300 Gaschromatograph tted with FID detector, split/splitless injector and a SPB-1 capillary column (30 m 032 mm ID, 025 lm lm thickness; Supelco, Bellefonte, PA, USA). Statistical analysis The results are expressed as mean SE, unless otherwise indicated. Data related to the baseline and to the Synbiotic and Placebo consumption periods were treated separately; results assessed at the different time points were also submitted to repeated measures analysis of variance, followed by Honesty Signicant Difference test. Univariate relationship between dietary intake, faecal microbial counts and results of biochemical analysis conducted on faecal samples were evaluated by Pearsons regression analysis, pooling together the data from the different experimental periods. Analysis were performed using the
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subjects belonging to the two arms of treatment (Table 1). During the treatment period, subjects consumed 500 ml day)1 of Synbiotic or Placebo in addition to their habitual diet. They followed no other dietary restrictions as, according to functional food denition, the Synbiotic must demonstrates their effects as a part of a normal food patterns (Diplock et al. 1999). To estimate their food intake during the trial, volunteers were asked to ll a food record in the last 3 days of each period (i.e. at the baseline start and end, T0d and T14d, and at the end, T42d, of Synbiotic or Placebo periods). Nutrient calculations were performed using the Italian Food Composition Tables (Carnovale and Marletta 1997). Intestinal habits were estimated using a self-administered questionnaire: after each defecation subjects answered questions (in a linear scale) about the ease of defecation, faeces consistency, abdominal pain, atulence and general well-being. At T0d, T14d and T42d, volunteers were asked to collect in a sterile plastic pot the rst stool passed in the morning. The samples were immediately introduced in anaerobic bags (Generbag Anaer, bioMerieux), cooled at +4C and delivered to the laboratory within 2 h from defecation for the microbiological and biochemical analyses. Faecal microbiological analyses Faecal sample analyses were performed under strict anaerobic conditions in an anaerobic cabinet (Forma Scientic, Marietta, OH, USA) under an atmosphere of N2/H2/CO2 (85/10/5, v/v/v). After homogenization, a subsample of 5 g was suspended (nal volume of 50 ml) in reduced dilution blank (Holdeman et al. 1977) and serial 10-fold dilutions were performed. The following microbial groups were enumerated using selective growth media: total anaerobes on blood agar (Holdeman et al. 1977); total aerobes and facultative anaerobes on Difco (Detroit, MI, USA) tryptic soy agar; coliforms on Difco MacConkey agar; clostridia on sultecolistinmilk agar (MevissenVerhage et al. 1982) after ethanol treatment of the dilutions; bidobacteria on Beerens medium (Beerens 1990); lactobacilli on Lamvab medium (Hartemink et al. 1997)

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statsoft Statistical Package for Windows (release 45; StatSoft Inc., Tulsa, OK, USA). Results All the volunteers completed each limb of the study: 500 ml day)1 of Placebo or Synbiotic was well accepted. One subject did not ll the food record at baseline so that nutrient intakes in this period were evaluated as a mean of 25 values. Baseline period slightly changed food habits of involved subjects, particularly a signicant increase in Ca (P < 0001), P (P < 005) and proteins (P < 005) intakes (Table 2) was assessed. The observed dietary changes, however, did not modify intestinal habits (data not shown). Furthermore, at T14d no signicant differences in dietary intakes were observed among subjects allocated in the two groups of subsequent treatment with Synbiotic or Placebo. During these dietary periods, nutrient intake was unaffected by treatments (Table 3). On the contrary, intestinal habits were modied by dietary treatments: Synbiotic consumption induced a slight amelioration of the easiness of defecation (149 105% vs 185.0 194% of the maximal defecation stress), and a signicant increase in atulence (120 180% vs

Table 4 Microbial counts in faecal samples of the subjects at the start (T0d) and the end (T14d) of the baseline period
Microbial groups (log CFU g)1 d.w.) Facultative anaerobes Anaerobes Bidobacteria Clostridia Coliforms Lactobacilli Lactobacilli T0d (n 26) 809 1082 993 458 745 608 n.d. 019 011 011 168 024 031 T14d (n 26) 828 1149 1051 459 768 637 641 017 006 018** 125 018 036 039

Values are presented as mean SE. n.d. not detected. Lactobacilli determined on Lamvab medium. Lactobacilli determined on Oxoid Rogosa agar medium. **P < 0001.

Table 2 Nutrient daily intakes at the start (T0d) and the end (T14d) of the baseline period
T0d (n 25) Energy (kJ) Protein (g) Fat (g) Carbohydrate (g) Dietary bre (g) Ca (mg) P (mg) Fe (mg) 80300 707 722 2488 176 9290 12097 101 4057 36 41 143 12 668 731 07 T14d (n 26) 79448 775 677 2441 146 11228 13477 105 2988 30* 37 107 11 578** 534* 12

Values are presented as mean SE. *P < 005; **P < 0001.

320 290% of the extreme sensation of discomfort; P < 005) without inducing abdominal pain. During baseline period (T0dT14d) we observed a signicant increase in bidobacteria counts (P < 0001) (Table 4). The other tested microbial groups did not show any signicant change. Table 5 shows results about faecal microbial counts during Placebo and Synbiotic treatment. At T14d no signicant differences were observed in faecal microbiota composition between the two groups of subjects. The consumption of Synbiotic for 4 weeks markedly modied faecal microbiota composition with a signicant increase in bidobacteria (P < 005) and in lactobacilli on Rogosa agar medium (P < 0001). Notably, an increase in the number of lactobacilli was not observed on Lamvab medium, where the presence of vancomycin inhibited the growth of the L. acidophilus strain added in the Synbiotic product. The faecal microbiota composition of volunteers who consumed 2% fat milk (placebo) did not show any change between T14d and T42d. Results about pH, dry weight, l-lactate and faecal bile acids during baseline and treatment period are shown in Tables 6 and 7, respectively. We observed a signicant increase (P < 005) in faecal pH between T0d and T14d. In

Placebo (n 13) T14d Energy (kJ) Protein (g) Fat (g) Carbohydrate (g) Dietary bre (g) Ca (mg) P (mg) Fe (mg) 79887 765 653 2551 157 11133 13165 101 4557 37 51 176 19 682 648 08 T42d 86968 799 759 2689 165 11651 14964 105 5895 33 47 220 17 714 890 07

Synbiotic (n 13) T14d 79009 785 701 2331 136 11324 13789 109 4049 49 55 122 12 963 866 24 T42d 79276 718 704 2471 148 9759 12382 102 5895 32 43 170 12 798 684 15

Table 3 Nutrient daily intakes at the start (T14d) and the end (T42d) of the Placebo and Synbiotic treatment periods

Values are presented as mean SE.

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Table 5 Microbial counts in faecal samples of the subjects at the start (T14d) and the end (T42d) of the Placebo and Synbiotic treatment periods

Microbial groups (log CFU g)1 d.w.) Facultative anaerobes Anaerobes Bidobacteria Clostridia Coliforms Lactobacilli Lactobacilli

Placebo (n 13) T14d 832 1149 1068 494 749 631 616 025 011 012 141 027 054 068 T42d 819 1131 1048 494 731 668 649 023 011 013 136 033 030 044

Synbiotic (n 13) T14d 824 1148 1037 424 786 642 662 025 007 032 082 026 049 047 T42d 773 1144 1076 361 728 665 823 022 012 022* 168 020 045 027**

Values are presented as mean SE. Lactobacilli determined on Lamvab medium. Lactobacilli determined on Oxoid Rogosa agar medium. *Signicantly different (P < 005) from T14d in Synbiotic group. **Signicantly different (P < 001) from T14d in Synbiotic group.

addition, a correlation between faecal pH and calcium consumption (P < 005; r 03406, n 72) was found. The other tested parameters did not change even if a trend to decrease in faecal concentration of secondary bile acids deoxycholic (3a,12a-dihydroxy-5b-cholanic acid, DCA) and lithocholic (3a-monohydroxy-5b-cholanic acid, LCA) was shown (Table 6). Unfortunately the two groups of treatment were not completely homogeneous for all the faecal biochemical
Table 6 Faecal biochemical parameters of the subjects at the start (T0d) and the end (T14d) of the baseline period
T0d (n 26) pH Dry weight (%) )1 L-Lactic acid (mg 100 g w.w.) Lithocholic acid (lg mg)1 w.w.) Deoxycholic acid (lg mg)1 w.w.) Total bile acids (lg mg)1 w.w.) Values are presented as mean SE. *P < 005. 687 203 1760 19 19 39 012 12 152 03 05 08 T14d (n 26) 705 255 1635 17 13 30 037* 61 658 02 02 04

variables at the start of the treatment period (T14d) (Table 7). In subjects belonging to the Placebo group, the faecal concentration of DCA was signicantly (P < 005) higher than that evaluated in the Synbiotic group. Synbiotic administration induced a signicant decrease in faecal l-lactic acid (P < 005) concentration. Furthermore, at the end of Synbiotic consumption period (T42d) faecal LCA concentration showed a low decrement; on the contrary, DCA concentration increased, although the differences were not statistically signicant due to the large variability among subjects. Bile acids correlated with bacterial counts. In particular, in the Synbiotic group LCA was negatively correlated with lactobacilli (on Lamvab medium P < 001, r )05405, n 29; on Rogosa agar P < 005, r )04814, n 19) and to bidobacteria (P < 005, r )03840, n 30). Discussion In the last decade, dairy industry has been quickly renewed to search for products characterized not only by

Table 7 Faecal biochemical parameters of the subjects at the start (T14d) and the end (T42d) of the Placebo and Synbiotic treatment periods

Placebo (n 13) T14d pH 709 Dry weight (%) 232 )1 L-Lactic acid (mg 100 g w.w.) 1827 Lithocholic acid (lg mg)1 w.w.) 157 Deoxycholic acid (lg mg)1 w.w.) 177 Total bile acids (lg mg)1 w.w.) 337 T42d 013 705 18 255 237 1635 022 182 141a 172 062 356

Synbiotic (n 13) T14d 010 711 17 235 183 2341 033 174 058ab 082 088 262 T42d 008 692 11 198 288 1545 035 129 018b 116 049 246 016 14** 297* 027 029ab 050

Values are presented as mean SE. *Signicantly different from T14d (P < 005) for synbiotic. **Signicantly different from T42d (P < 001) for synbiotic. Values in the same row with different superscript are signicantly different (P < 005).
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their elevated nutritional value and pleasant taste, but also by their ability to exert positive effects on the consumer health. In the present study, the effect of a synbiotic milk-based product was tested on human volunteers. In accordance with PASSCLAIM consensus Criterion 2 (Agget et al. 2005), we characterized the background diet of the study groups. Dietary intervention study induced modications of intestinal ecosystem. In the baseline period, during which all the volunteers consumed 500 ml day)1 of skimmed milk for 2 weeks, an increase in the intake of protein, calcium and phosphorus was observed, probably due to the consumption of milk, rich in these elements, in quantity higher than usual. Moreover, in the same period a signicant increase of faecal pH was evidenced. This result can be partially related to the high level of calcium intake assessed in the same period, as suggested by the direct correlation between faecal pH and calcium consumption. Similarly, Govers et al. (1996) reported an increase of faecal pH, after administration of a milk product, ascribed to the buffered action of calcium and phosphorus. Regarding faecal microbiota composition, the Placebo administration during baseline induced a signicant increase in bidobacteria counts. Some authors (Geesey et al. 2000) evidenced that calcium can inuence the adhesion ability of bacteria, and Kim (1988) found that high calcium levels induced in vitro an increase in the adhesiveness of various bidobacteria species. Therefore, it is possible that variations in the mucosa adhesion ability, calcium mediated, have played an important role in the regulation of the bidobacteria indigenous populations during our experiment. Moreover, some authors found bidobacteria growth-promoting factors in bovine milk (Etienne et al. 1994; Brody 2000). It is also possible that an increased lactose ux in the colon, as a consequence of consumption of 500 ml of bovine milk per day, could have contributed to the bidogenic effect assessed during baseline period. Nowadays no published data support the bidogenic effect of lactose. Thus, this is an important result suggesting that milk consumption can favour the development of useful micro-organisms, particularly bidobacteria, in subjects not accustomed to high daily intake of such a food. Moreover, during baseline period faecal LCA and DCA concentrations decreased although not signicantly. As known, colonic bacteria are involved in intestinal bile acid metabolism: the conjugated primary cholic (3a,7a,12a-trihydroxy-5-b-cholanic acid) and chenodeoxycholic acids (3a,7a-dihydroxy-5-b-cholanic acid) that escaped the absorption in the last part of the ileum are deconjugated and 7a-dehydroxylated in the secondary DCA and LCA respectively by colonic bacteria. Consequently, the observed trend could be due to a lower 7a-dehydroxylating activity probably due to a reduction in the counts of
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microbial populations able to perform the conversion of primary to secondary bile acids. After the following treatment period of 4 weeks, during which the two groups of volunteers consumed 500 ml day)1 of the Synbiotic or of the Placebo, no differences in dietary intakes were estimated, but intestinal habits were modied by dietary treatments: Synbiotic consumption induced a slight amelioration of the ease of defecation and an increase in atulence, without inducing abdominal pain. Other authors have recorded an increase of gas production during inulin administration, sometimes with intestinal troubles which were attributed either to inulin intake or to individual sensitivity (Alles et al. 1996, 1999; Pedersen et al. 1997; Davidson et al. 1998). Moreover we observed a reduction, although nonsignicant, of faecal dry weight in Synbiotic-treated subjects which could be associated with the assessed increased facility of defecation. We did not observe faeces acidication in the Synbiotic-treated group that may be related to the milk matrix, rich in buffering substances; moreover, as suggested by other authors (Alles et al. 1996; Bouhnik et al. 1996), the short-chain organic acids, produced during inulin fermentation, could be metabolized faster by the colonic mucosa. Synbiotic consumption also affected intestinal microbiota composition. In subjects treated with the Synbiotic we assessed an increase in bidobacteria and lactobacilli counts; on the contrary, Placebo did not modify faecal microbial composition. Notably, faecal l-lactic acid concentration decreased signicantly (P < 005) after Synbiotic consumption, suggesting a stimulation of the growth of lactate-utilizer microbial groups. It should be interesting to test this hypothesis as some genera, such as propionibacteria, could be considered useful (Grant and Salminen 1998), but others, particularly sulfate-reducing bacteria producing cytotoxic H2S, are dangerous for the host. Our data did not show any signicant change in faecal concentration of secondary bile acids during treatment period even if LCA tends to decrease while DCA tends to increase after Synbiotic administration. In addition, in the same period we evidenced a signicant inverse correlation between faecal concentration of LCA and bidobacteria and lactobacilli counts. It is likely that a lower pH, probably induced in the colon by such lactic acid-producing bacteria and by inulin fermentation, could have partially inhibited the 7a-dehydroxylating activity, thus reducing LCA production. Notably this secondary bile acid, poorly soluble, is mostly excreted in faeces. On the contrary, faecal DCA concentration tend to increase. This result is an apparent contrast to the above hypothesis. In fact, the lower pH could reduce the DCA solubility increasing its

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precipitation and, if this is the case, the lower production of DCA could be hidden by its higher faecal concentration. The above-suggested mechanism could modulate entero-hepatic circulation of bile acids; particularly a reduced absorption of secondary bile acids should stimulate the reconstitution of bile acid pool from endogenous cholesterol, with a potential reduction of plasma cholesterol level. However it must be noted that the methodological approach of this study do not allow us to assess the role of inulin and probiotics separately with respect to the modulation of colonic ecosystem and consequently bile acid metabolism. Actually, not many studies investigated the effect of inulin and probiotic bacteria in human bile acids metabolism, thus further studies are needed, to conrm our data and in particular to clarify the role of prebiotic and probiotic consumption on bile acid metabolism. We conclude that the consumption of the tested Synbiotic may positively modulate the intestinal ecosystem as it ameliorated intestinal habits and induced an increase in faecal lactobacilli and bidobacteria counts. References
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