Specific Response of a Novel and Abundant Lactobacillus amylovorus-Like Phylotype to Dietary Prebiotics in the Guts of Weaning Piglets

Laboratory of Microbiology, Agrotechnology and Food Sciences Group, Wageningen University, Hesselink van Suchtelenweg 4, 6703 CT Wageningen, The Netherlands.
Applied and Environmental Microbiology (Impact Factor: 3.67). 08/2004; 70(7):3821-30. DOI: 10.1128/AEM.70.7.3821-3830.2004
Source: PubMed


Using 16S rRNA gene-based approaches, we analyzed the responses of ileal and colonic bacterial communities of weaning piglets to dietary addition of four fermentable carbohydrates (inulin, lactulose, wheat starch, and sugar beet pulp). An enriched diet and a control diet lacking these fermentable carbohydrates were fed to piglets for 4 days (n = 48), and 10 days (n = 48), and the lumen-associated microbiota were compared using denaturing gradient gel electrophoresis (DGGE) analysis of amplified 16S rRNA genes. Bacterial diversities in the ileal and colonic samples were measured by assessing the number of DGGE bands and the Shannon index of diversity. A higher number of DGGE bands in the colon (24.2 +/- 5.5) than in the ileum (9.7 +/- 4.2) was observed in all samples. In addition, significantly higher diversity, as measured by DGGE fingerprint analysis, was detected in the colonic microbial community of weaning piglets fed the fermentable-carbohydrate-enriched diet for 10 days than in the control. Selected samples from the ileal and colonic lumens were also investigated using fluorescent in situ hybridization (FISH) and cloning and sequencing of the 16S rRNA gene. This revealed a prevalence of Lactobacillus reuteri in the ileum and Lactobacillus amylovorus-like populations in the ileum and the colon in the piglets fed with fermentable carbohydrates. Newly developed oligonucleotide probes targeting these phylotypes allowed their rapid detection and quantification in the ileum and colon by FISH. The results indicate that addition of fermentable carbohydrates supports the growth of specific lactobacilli in the ilea and colons of weaning piglets.

Download full-text


Available from: Barbara A Williams
  • Source
    • "A recent study showed that lactulose and Lactobacillus plantarum synbiotic treatment has positive effects in controlling colibacillosis in post-weaning pigs (Guerra-Ordaz et al., 2014). In addition, lactulose was shown to alter specific gut microflora in sow using the denaturation gradient gel electrophoresis method (Konstantinov et al., 2004a; Zheng et al., 2014). Although the study presented interesting results regarding its effects on animal gut physiology, morphology, and metabolites, the influence of lactulose on the composition of the gut microbiome requires further study. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Lactulose is a synthetic by-product of the isomerization of lactose to disaccharide galactosyl β-(1,4) fructose, and is widely applied as an oral therapeutic and laxative. As a prebiotic, lactulose is thought to control gastrointestinal diseases associated with weaning both in humans and animals. This study focuses on the effect of lactulose as a prebiotic supplement in modulating the microbial community composition and diversity in 30 piglets. The swine faecal microbiome was investigated using 16S rRNA gene barcoded-pyrosequencing before and after lactulose administration for 2 weeks. Major shifts in bacterial composition were detected from the phylum to species level. An increased abundance of Firmicutes phylum and lower abundance of the Bacteroidetes phylum were observed in pigs fed with lactulose compared with the control group. Interestingly, there was a negative relationship between lactulose administration and abundance of the genus Escherichia. In addition, a marked increase in the abundance of Lactobacillus reuteri (1.15% to 6.28%) was observed in the lactulose group. The canonical loading plot showed that three genera, Prevotella, Subdoligranulum, and Clostridium, were highly discriminating genera that explain the separation of swine faecal microbiota by lactulose administration. Overall, our results showed that lactulose administration could effectively increase diversity and create a distinct microbial community in weaned piglets, which is believed to be important and beneficial to health and performance.
    Full-text · Article · Jul 2015 · Animal Feed Science and Technology
  • Source
    • "However , the number of bands was consistent with what Liu et al.[14]observed in neonatal piglets. Potential reasons for the discrepancy between Konstanintov et al.[44]and the results of the present study and that of Liu et al.[14]were that the samples were collected at different ages and the piglets were fed different diets. Based on the findings reported above, it is evident that one mechanism through which L. reuteri I5007 functions is by increasing the presence of beneficial bacterial species in the lower gastrointestinal tract and decreasing the presence of undesirable microbes. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Seventy-two, suckling piglets, obtained from 9 litters standardized to 8 piglets, were assigned to 1 of 3 treatments (n = 24) to compare short-term, early administration with intermittent, longer-term administration of Lactobacillus reuteri I5007. The treatments were a control (given a placebo of 0.1% peptone water from day 1 to 5) or treatments in which 1.7 × 1010 CFU L. reuteri was administrated either daily for 4 days starting on day 1 or every 4th day from day 1 to 17. Five piglets per treatment were killed at 3 time points (day 7, 14 and 21). Denaturing Gradient Electrophoresis of ileal digesta revealed an increase in the presence of L. reuteri I5007 and Clostridium lentocellum (on day 14 and 21) in the every 4th-day treatment and Actinobacillus porcinus (on day 7 and 14) in both L. reuteri treatments, while reducing the abundance of E. coli on day 21 in the every 4th-day treatment. Real-time qPCR of ileal digesta showed an increase in Bifidobacterium spp. on day 14 for both L. reuteri I5007 treatments. An increase in the concentration of lactic acid and a lower pH was observed in the first 4-day treatment on day 7 and the every 4th day treatment on day 14. The relative abundance of mRNA for TGF-β was increased while that for IFN-γ was decreased in the mesenteric lymph nodes of piglets treated with L. reuteri every 4th day. In conclusion, early intervention with L. reuteri increases the presence of beneficial bacteria and decreases the presence of undesirable microbes in the lower gastrointestinal tract. The changes appear to be mediated by altering the intestinal pH through lactic acid production resulting in favorable bacterial species colonization. A prolonged duration of treatment (i.e. every 4th day) would appear to be superior to treatment only during the first 4 days.
    Full-text · Article · Mar 2015 · PLoS ONE
  • Source
    • "Previous studies showed that up to 37% of the pectin-rich fraction in SBP is already fermented before the ileum, and another 50% disappeared in the large intestine of pigs (Graham et al. 1986). Konstantinov et al. (2004) determined a considerable microbial activity and increase in lactobacilli populations in the pig small intestine when diets containing SBP were fed. In the stomach, SBP increased SCFA production and a shift from acetate towards propionate and butyrate, showing the relevance of bacterial fermentation in this part of the porcine GIT. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Although fermentable carbohydrates (CHO) can reduce metabolites derived from dietary protein fermentation in the intestine of pigs, the interaction between site of fermentation and substrate availability along the gut is still unclear. The current study aimed at determining the impact of two different sources of carbohydrates in diets with low or very high protein content on microbial metabolite profiles along the gastrointestinal tract of piglets. Thirty-six piglets (n = 6 per group) were fed diets high (26%, HP) or low (18%, LP) in dietary protein and with or without two different sources of carbohydrates (12% sugar beet pulp, SBP, or 8% lignocellulose, LNC) in a 2 × 3 factorial design. After 3 weeks, contents from stomach, jejunum, ileum, caecum, proximal and distal colon were taken and analysed for major bacterial metabolites (D-lactate, L-lactate, short chain fatty acids, ammonia, amines, phenols and indols). Results indicate considerable fermentation of CHO and protein already in the stomach. HP diets increased the formation of ammonia, amines, phenolic and indolic compounds throughout the different parts of the intestine with most pronounced effects in the distal colon. Dietary SBP inclusion in LP diets favoured the formation of cadaverine in the proximal parts of the intestine. SBP mainly increased CHO-derived metabolites such as SCFA and lactate and decreased protein-derived metabolites in the large intestine. Based on metabolite profiles, LNC was partly fermented in the distal large intestine and reduced mainly phenols, indols and cadaverine, but not ammonia. Multivariate analysis confirmed more diet-specific metabolite patterns in the stomach, whereas the CHO addition was the main determinant in the caecum and proximal colon. The protein level mainly influenced the metabolite patterns in the distal colon. The results confirm the importance of CHO source to influence the formation of metabolites derived from protein fermentation along the intestinal tract of the pig.
    Full-text · Article · Jun 2014 · Archives of animal nutrition
Show more