PCR and real-time PCR primers developed for detection and identification of Bifidobacterium thermophilumin faeces

Mathys, Sophie; Lacroix, Christophe; Mini, Raffaella; Meile, Léo

doi:10.1186/1471-2180-8-179

Cited by 19 publications

(13 citation statements)

References 37 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…B. gallicum was mostly absent (15,30,33), except for a baby delivered by caesarean section (6). B. thermophilum, first isolated from baby feces (53), was recently detected in the feces of a breastfed infant at 6.7 log 10 cells/g feces (28). Since B. pseudolongum has so far been associated with animals, it has rarely been investigated in humans.…”

mentioning

confidence: 99%

Quantification of Human Fecal Bifidobacterium Species by Use of Quantitative Real-Time PCR Analysis Targeting the groEL Gene

Junick

Blaut

2012

Appl Environ Microbiol

106

View full text Add to dashboard Cite

ABSTRACTQuantitative real-time PCR assays targeting thegroELgene for the specific enumeration of 12 human fecalBifidobacteriumspecies were developed. The housekeeping genegroEL(HSP60in eukaryotes) was used as a discriminative marker for the differentiation ofBifidobacterium adolescentis,B. angulatum,B. animalis,B. bifidum,B. breve,B. catenulatum,B. dentium,B. gallicum,B. longum,B. pseudocatenulatum,B. pseudolongum, andB. thermophilum. The bifidobacterial chromosome contains a single copy of thegroELgene, allowing the determination of the cell number by quantification of thegroELcopy number. Real-time PCR assays were validated by comparing fecal samples spiked with known numbers of a givenBifidobacteriumspecies. Independent of theBifidobacteriumspecies tested, the proportion ofgroELcopies recovered from fecal samples spiked with 5 to 9 log10cells/g feces was approximately 50%. The quantification limit was 5 to 6 log10groELcopies/g feces. The interassay variability was less than 10%, and variability between different DNA extractions was less than 23%. The method developed was applied to fecal samples from healthy adults and full-term breast-fed infants. Bifidobacterial diversity in both adults and infants was low, with mostly ≤3Bifidobacteriumspecies andB. longumfrequently detected. The predominant species in infant and adult fecal samples wereB. breveandB. adolescentis, respectively. It was possible to distinguishB. catenulatumandB. pseudocatenulatum. We conclude that thegroELgene is a suitable molecular marker for the specific and accurate quantification of human fecalBifidobacteriumspecies by real-time PCR.

show abstract

mentioning

confidence: 99%

Quantification of Human Fecal Bifidobacterium Species by Use of Quantitative Real-Time PCR Analysis Targeting the groEL Gene

Junick

Blaut

2012

Appl Environ Microbiol

106

View full text Add to dashboard Cite

show abstract

“…Standard curves for each target group were prepared as described previously (28) , and assays were carried out using the KAPA SYBR ® FAST qPCR Kit (Kapa Biosystems, Inc.) in a reaction volume of 25 μl. A TaqMan assay was carried out for the enumeration of B. thermophilum with specific primers (online Supplementary Table S2) using the RT-QP2X-03WOULR Mastermix (Eurogentec S.A.) and reaction conditions set by Mathys et al (6) .…”

Section: Microbial Composition By Quantitative Polymerase Chain Reactionmentioning

confidence: 99%

“…With safe history of use, strains of the genera Bifidobacterium and Lactobacillus are widely represented among probiotics (2) . Bifidobacterium thermophilum strains have been primarily isolated from the bovine rumen and animal faeces, mainly calf and pig (3,4) , and have also been found in the faeces of babies (5,6) . The probiotic potential of the human faecal isolate B. thermophilum RBL67 (RBL67) was investigated in several in vitro studies.…”

mentioning

confidence: 99%

Effect of Bifidobacterium thermophilum RBL67 and fructo-oligosaccharides on the gut microbiota in Göttingen minipigs

Tanner

Lacroix

Del’Homme³

et al. 2015

Br J Nutr

Self Cite

View full text Add to dashboard Cite

Modulating the gut microbiota via dietary interventions is a common strategy to enhance the natural defence mechanisms of the host. Several in vitro studies have highlighted the probiotic potential of Bifidobacterium thermophilum RBL67 (RBL67) selected for its anti-Salmonella effects. The present study aimed to investigate the impact of RBL67 alone and combined with fructooligosaccharides (FOS) on the gut microbiota of Göttingen minipigs. Minipigs were fed a basal diet supplemented with 8 g/d probiotic powder (1 × 10 9 CFU/g in skim milk matrix) (probiotic diet (PRO)), 8 g/d probiotic powder plus 8 g/d FOS (synbiotic diet (SYN)) or 8 g/d skim milk powder (control), following a cross-sectional study design. Faecal and caecal microbiota compositions were analysed with pyrosequencing of 16S rRNA genes and quantitative PCR. Metabolic activity in the caecum and colon was measured by HPLC. 16S rRNA gene amplicon sequencing revealed that minipig faeces show close similarity to pig microbiota. During the treatments and at the time of killing of animals, RBL67 was consistently detected in faeces, caecum and colon at numbers of 10 5 -10 6 16S rRNA copies/g content after feeding PRO and SYN diets. At the time of killing of animals, significantly higher Bifidobacterium numbers in the caecum and colon of SYN-fed minipigs were measured compared with PRO. Our data indicate that the Göttingen minipig may be a suitable model for gut microbiota research in pigs. Data from this first in vivo study of RBL67 colonisation suggest that the combination with FOS may represent a valuable symbiotic strategy to increase probiotic bacteria levels and survival in gastrointestinal tracts for feed and food applications.Key words: Probiotics: Synbiotics: Gut microbiota modulation: In vivo studies: Animal modelsThe gut microbiota provides a beneficial barrier function for the host, especially against common enteropathogens such as Salmonella or Escherichia coli that are encountered in pig farms. This barrier effect is possibly enhanced by probiotic bacteria that contribute to gut microbiota modulation. Specific characteristics attributed to probiotics include competition for nutrients and adhesion sites, stimulation of SCFA, production of antimicrobial substances and modulation of the immune response (1,2) . Probiotic traits are strain-specific, and must therefore be proven for any given strain. With safe history of use, strains of the genera Bifidobacterium and Lactobacillus are widely represented among probiotics (2) . Bifidobacterium thermophilum strains have been primarily isolated from the bovine rumen and animal faeces, mainly calf and pig (3,4) , and have also been found in the faeces of babies (5,6) . The probiotic potential of the human faecal isolate B. thermophilum RBL67 (RBL67) was investigated in several in vitro studies. Strain RBL67 produces a bacteriocin-like substance (5,7) , adheres to human intestinal cell lines (8) and exhibits protective effects on epithelial integrity in the presence of a competing human microbiota (9) . Fu...

show abstract

“…thermophilum enumeration was performed using primers bthermRTF and bthermRTR and the Taqman probe bthermTqm (Additional file 3: Table S1) [60]. The RT-QP2X-03WOULR Mastermix (Eurogentec s.a., Seraing, Belgium) was used and standard curve preparation and reaction conditions were carried out as described previously [59,60].…”

Section: Qpcr Analysesmentioning

confidence: 99%

Synergistic effects of Bifidobacterium thermophilum RBL67 and selected prebiotics on inhibition of Salmonella colonization in the swine proximal colon PolyFermS model

et al. 2014

Self Cite

View full text Add to dashboard Cite

Background: Probiotics and prebiotics are promising strategies to counteract Salmonella prevalence in swine. In the present study, we investigated the effects of prebiotics (fructo-(FOS), galacto-(GOS) and mannan-(MOS) oligosaccharides) and the bacteriocinogenic Bifidobacterium thermophilum RBL67 (RBL67) on Salmonella enterica subsp. enterica serovar Typhimurium N-15 (N-15) colonization using the PolyFermS in vitro continuous fermentation model simulating the swine proximal colon. Material and methods: The PolyFermS model was designed with a first-stage reactor containing immobilized fecal pig microbiota. This reactor continuously inoculated five parallel second-stage reactors, a control and four treatment reactors, all operated with proximal colon conditions. FOS and GOS (5.2 g/day), and MOS (half dosage) and RBL67 (10 8 copy numbers/mL applied daily) were tested on the ability of N-15 to colonize reactors, inoculated with the same microbiota. Reactor effluents were collected daily and analyzed for microbial composition (quantitative PCR and 454 pyrosequencing of 16S rRNA gene pool) and main metabolites (HPLC).

show abstract

PCR and real-time PCR primers developed for detection and identification of Bifidobacterium thermophilumin faeces

Cited by 19 publications

References 37 publications

Quantification of Human Fecal Bifidobacterium Species by Use of Quantitative Real-Time PCR Analysis Targeting the groEL Gene

Quantification of Human Fecal Bifidobacterium Species by Use of Quantitative Real-Time PCR Analysis Targeting the groEL Gene

Effect of Bifidobacterium thermophilum RBL67 and fructo-oligosaccharides on the gut microbiota in Göttingen minipigs

Synergistic effects of Bifidobacterium thermophilum RBL67 and selected prebiotics on inhibition of Salmonella colonization in the swine proximal colon PolyFermS model

Contact Info

Product

Resources

About