Discrimination between Bifidobacterium Species from Human and Animal Origin by PCR–Restriction Fragment Length Polymorphism

Delcenserie, Véronique; Bechoux, Nathalie; Leonard, Tamara; China, Bernard; Daube, Georges

doi:10.4315/0362-028x-67.6.1284

Cited by 22 publications

(21 citation statements)

References 18 publications

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“…In addition, three more combined patterns were observed along the cheese process: II-IX (presumed human origin bifidobacteria [23], V-IX and V-X. One hundred and eight samples (61%) were V-X type positive and 31 (18%) were V-IX type positive.…”

Section: Resultsmentioning

confidence: 99%

“…The enrichment medium was Brain Heart Infusion (BHI, 37 g l -1 , Bio-Rad, Marnes-la-Coquette, France), supplemented with several components (propionic acid, 5 ml l -1 ; Fe-citrate, 0.5 g l -1 ; cystein chlorhydrate, 0.5 g l -1 ; yeast extract, 5 g l -1 ; agar, 2 g l -1 ) and mupirocin (Lithium mupirocin, GlaxoSmithKline, England) as the selective agent at a final concentration of 80 mg l -1 [23]. …”

Section: Methodsmentioning

confidence: 99%

“…A 1050 bp amplicon of the 16S rDNA gene was generated using primers: 16S up: 5'-AAT AGC TCC TGG AAA CGG GT-3' and 16S down: 5'-CGT AAG GGG CAT GAT GAT CT-3' (Eurogentec, Seraing, Belgium; Genbank PUID: updown16S EOY_1) as previously described [23]. The digestion of the PCR products for species detection was performed using two enzymes: Alu I and Taq I (Roche; Basel, Switzerland) as described previously [23]. Following the digestion, the products were analyzed by gel electrophoresis using 2.5% agarose gel.…”

Section: Methodsmentioning

confidence: 99%

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Bifidobacterium pseudolongum are efficient indicators of animal fecal contamination in raw milk cheese industry

Delcenserie

Gavini²,

China

et al. 2011

BMC Microbiol

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BackgroundThe contamination of raw milk cheeses (St-Marcellin and Brie) from two plants in France was studied at several steps of production (raw milk, after addition of rennet - St-Marcellin - or after second maturation - Brie -, after removal from the mold and during ripening) using bifidobacteria as indicators of fecal contamination.ResultsBifidobacterium semi-quantitative counts were compared using PCR-RFLP and real-time PCR. B. pseudolongum were detected in 77% (PCR-RFLP; 1.75 to 2.29 log cfu ml-1) and 68% (real-time PCR; 2.19 to 2.73 log cfu ml-1) of St-Marcellin samples and in 87% (PCR-RFLP; 1.17 to 2.40 log cfu ml-1) of Brie cheeses samples. Mean counts of B. pseudolongum remained stable along both processes. Two other populations of bifidobacteria were detected during the ripening stage of St-Marcellin, respectively in 61% and 18% of the samples (PCR-RFLP). The presence of these populations explains the increase in total bifidobacteria observed during ripening. Further characterization of these populations is currently under process. Forty-eight percents (St-Marcellin) and 70% (Brie) of the samples were B. pseudolongum positive/E. coli negative while only 10% (St-Marcellin) and 3% (Brie) were B. pseudolongum negative/E. coli positive.ConclusionsThe increase of total bifidobacteria during ripening in Marcellin's process does not allow their use as fecal indicator. The presence of B. pseudolongum along the processes defined a contamination from animal origin since this species is predominant in cow dung and has never been isolated in human feces. B. pseudolongum was more sensitive as an indicator than E. coli along the two different cheese processes. B. pseudolongum should be used as fecal indicator rather than E. coli to assess the quality of raw milk and raw milk cheeses.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Bifidobacterium pseudolongum are efficient indicators of animal fecal contamination in raw milk cheese industry

Delcenserie

Gavini²,

China

et al. 2011

BMC Microbiol

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“…[54] The "bifid shunt" generates acetyl phosphate and erythrose 4-phosphate. The final product of the fermentation generates glyceraldehyde 3-phosphate that further enters to the Embden-MeyerhofParnas pathway.…”

Section: Role Of Probiotics In Diabetes Managementmentioning

confidence: 99%

Untitled

Prabhakar¹

2017

IJGP

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Diabetes mellitus is commonly known worldwide by the name of diabetes which occurs in all age groups. About >90% of diagnosed patient specially are Type-2 diabetes. The hallmark of Type 2 diabetes developments is increased insulin resistance, whereas Type 1 is related to less production of insulin which leads to uncontrolled hyperglycemia. Hyperglycemia slowly produces mild-to-very serious complications in patient mainly affecting vital organs such as blood vessels, eyes, neurons, nephrons, heart, and brain which increase the risk of heart attack, retinopathy, nephropathy, neuropathy, and stroke. Proper management of hyperglycemia is a key to prevent from diabetes and its complications. This concept has attracted many researchers to target various cells and tissue through special remedy so that hyperglycemia can be managed and complications can be reduced. The patient shows numerous side effects during therapy. There is craving demand for the proper cure of diabetes by sufferers. In this aspect, probiotics can be more helpful if proper research and formulation are done. Probiotics are good microorganism which can control hyperglycemia and its complications by utilizing and modifying glucose before absorption. Appropriate research is required to make strategy for searching and formulating good microorganism to be used as probiotics for the regulation of blood glucose and prevention from complexity.

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“…Several molecular methods that alleviate this inconvenience have recently been described: PCR-Elisa method based on the 16S rRNA to detect the most common Bifidobacterium species in humans (Malinen et al, 2002); pulsed-field gel electrophoresis (PFGE) and PCR targeting the 16S rRNA (Roy et al, 1996;Bonjoch et al, 2004); PCR in denaturing gradient gel electrophoresis (DGGE) targeting the transaldolase gene for identification, detection and enumeration of human Bifidobacterium species (Requena et al, 2002); PCR-RFLP method based on the 16SrRNA to detect the most common species from animal and human origins (Delcenserie et al, 2004;Roy and Sirois, 2000), and real-time quantitative PCR from the 16S or the transaldolase gene (Requena et al, 2002). They have also been used in the detection of human Bifidobacterium species from feces Requena et al, 2002, Mullié et al, 2003Venema and Maathuis, 2003), of bifidobacteria as probiotics (Brigidi et al, 2003;Fasoli et al, 2003) or as fecal indicators in waters (Bernhard and Field, 2000).…”

Section: Introductionmentioning

confidence: 99%

A PCR method for detection of bifidobacteria in raw milk and raw milk cheese: comparison with culture-based methods

Delcenserie

Bechoux

China

et al. 2005

Journal of Microbiological Methods

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Bifidobacteria are well known for their beneficial effects on health and are used as probiotics in food and pharmaceutical products. As they form one of the most important groups in both human and animal feces, their use as fecal indicator organisms in raw milk products has recently been proposed. Bifidobacteria species isolated in humans are different from those isolated in animals. It should therefore be possible to determine contamination origin (human or animal).A method of detecting the Bifidobacterium genus was developed by PCR targeting the hsp60 gene. The genus Bifidobacterium was identified by PCR amplification of a 217-bp hsp60 gene fragment. The degenerated primer pair specific to the Bifidobacterium genus used was tested for it specificity on 127 strains. Sensitivity was measured on artificially contaminated samples. Food can however be a difficult matrix for PCR testing since it contains PCR inhibitors. So an internal PCR control was used. An artificially created DNA fragment of 315 bp was constructed. The PCR detection method was tested on raw milk and cheese samples and compared with three culture-based methods, which comprised enrichment and isolation steps. The enrichment step used Brain Heart Infusion medium with propionic acid, iron citrate, yeast extract, supplemented with mupirocin (BHMup) or not (BH) and the isolation step used Columbia blood agar medium, supplemented with mupirocin (CMup) or not (C). The method using mupirocin at both enrichment and isolation steps and the PCR method performed from the culture in BHMup enrichment medium were shown to be the most efficient. No significant difference was observed in raw milk samples between PCR from BHMup and the culture-based method BHMup/CMup, while a significant difference was noticed between the same methods in raw milk cheese samples, which would favor using PCR.The results suggested that PCR on the hsp60 gene was convenient for a rapid detection of bifidobacteria in raw milk and raw milk cheese samples and that bifidobacteria always present throughout raw milk cheese production could be efficiently used as fecal indicators.

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Discrimination between Bifidobacterium Species from Human and Animal Origin by PCR–Restriction Fragment Length Polymorphism

Cited by 22 publications

References 18 publications

Bifidobacterium pseudolongum are efficient indicators of animal fecal contamination in raw milk cheese industry

Bifidobacterium pseudolongum are efficient indicators of animal fecal contamination in raw milk cheese industry

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A PCR method for detection of bifidobacteria in raw milk and raw milk cheese: comparison with culture-based methods

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