Distribution of lactobacilli in the porcine gastrointestinal tract

The ability of Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus administered in yogurt to survive the passage through the upper gastrointestinal tract was investigated with Göttingen minipigs that were fitted with ileum T-cannulas. After ingestion of yogurt containing viable microorganisms, ileostomy samples were collected nearly every hour beginning 3 h after food uptake. Living L. delbrueckii subsp. bulgaricus and S. thermophilus were detected in the magnitude of 10 6 to 10 7 per gram of intestinal contents (wet weight) in all animals under investigation. A calculation of the minimum amount of surviving bacteria that had been administered is presented. Total DNA extracted from ileostomy samples was subjected to PCR, which was species specific for L. delbrueckii and S. thermophilus and subspecies specific for L. delbrueckii subsp. bulgaricus. All three bacterial groups could be detected by PCR after yogurt uptake but not after uptake of a semisynthetic diet. One pig apparently had developed an endogenous L. delbrueckii flora. When heat-treated yogurt was administered, L. delbrueckii was detected in all animals. S. thermophilus or L. delbrueckii subsp. bulgaricus was not detected, indicating that heat-inactivated cells and their DNAs had already been digested and their own L. delbrueckii flora had been stimulated for growth.

Section: Methodsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Survival of Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus in the Terminal Ileum of Fistulated Göttingen Minipigs

Lick¹,

Drescher²,

Heller³

2001

“…While, these Lactobacillus populations establish early in the piglet, succession occurs throughout the pig's lifetime, with lactobacilli remaining a predominant portion of the population (29,31). Several reports indicate that lactobacilli exert antagonistic characteristics toward other bacteria and fungal species (6). Numerous species and strains of Lactobacillus, including L. reuteri, have been detected and isolated from pig intestine and feces using both conventional and molecular techniques (2,17,24).…”

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confidence: 99%

“…Currently, methods based on genes encoding 16S rRNA (16S rDNA) are being used to examine differences in the bacteria inhabiting the gastrointestinal tract of a variety of animal models, such as mice (15,38), rabbits (39), chickens (19), pigs (17,21,22,31), and humans (14,33,40). At present, molecular analysis of the porcine gastrointestinal microbiota has been limited to examination of samples that were cultivated prior to genotypic analysis (2,6,11,22,31,34).…”

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confidence: 99%

Denaturing Gradient Gel Electrophoresis Analysis of 16S Ribosomal DNA Amplicons To Monitor Changes in Fecal Bacterial Populations of Weaning Pigs after Introduction of Lactobacillus reuteri Strain MM53

Simpson

McCracken

Gaskins

et al. 2000

172

124

The diversity and stability of the fecal bacterial microbiota in weaning pigs was studied after introduction of an exogenous Lactobacillus reuteri strain, MM53, using a combination of cultivation and techniques based on genes encoding 16S rRNA (16S rDNA). Piglets (n ‫؍‬ 9) were assigned to three treatment groups (control, daily dosed, and 4th-day dosed), and fresh fecal samples were collected daily. Dosed animals received 2.5 ؋ 10 10 CFU of antibiotic-resistant L. reuteri MM53 daily or every 4th day. Mean Lactobacillus counts for the three groups ranged from 1 ؋ 10 9 to 4 ؋ 10 9 CFU/g of feces. Enumeration of strain L. reuteri MM53 on MRS agar (Difco) plates containing streptomycin and rifampin showed that the introduced strain fluctuated between 8 ؋ 10 3 and 5 ؋ 10 6 CFU/g of feces in the two dosed groups. Denaturing gradient gel electrophoresis (DGGE) of PCR-amplified 16S rDNA fragments, with primers specific for variable regions 1 and 3 (V1 and V3), was used to profile complexity of fecal bacterial populations. Analysis of DGGE banding profiles indicated that each individual maintained a unique fecal bacterial population that was stable over time, suggesting a strong host influence. In addition, individual DGGE patterns could be separated into distinct time-dependent clusters. Primers designed specifically to restrict DGGE analysis to a select group of lactobacilli allowed examination of interspecies relationships and abundance. Based on relative band migration distance and sequence determination, L. reuteri was distinguishable within the V1 region 16S rDNA gene patterns. Daily fluctuations in specific bands within these profiles were observed, which revealed an antagonistic relationship between L. reuteri MM53 (band V1-3) and another indigenous Lactobacillus assemblage (band V1-6).

“…Prebiotics have been used to induce the colonization of bacteria, such as lactobacilli and bifidobacteria, considered to be beneficial for the host (13,14,61). Stimulation of the Lactobacillus population within the GI tracts of piglets is of specific importance, not only due to the potential effect of the bacteria on gut function and health (57,58), but also because of their possible antagonistic activities toward other bacteria (19,51). Lactobacilli establish early in the piglet intestine, and although succession occurs throughout the pig's lifetime, they remain a predominant part of the intestinal bacterial community (3,38,57,60).…”

mentioning

confidence: 99%

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

Konstantinov

Awati

Smidt

et al. 2004

195

159

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.