Differentiation of Enterococcus faecium from Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus strains by PCR and dot-blot hybridisation

Langa, Susana; Fernández, Antonio José Díaz; Martı́n, Rocı́o; Reviriego, C.; Marín, María; Fernández, Leónides; Rodríguez, Juan Miguel Díaz

doi:10.1016/s0168-1605(03)00180-6

Cited by 7 publications

(8 citation statements)

References 7 publications

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“…After isolation and biochemical characterisation of single colonies obtained from faecal samples at the 50 and 75% stages of the experimental period, PCR amplification of the 16S-23S rRNA gene spacer region with specific primers for E. faecium yielded two amplification products: one of 200 bp and the other of 300 bp. However, these results differ from those of Langa et al [17], who used the same primer combination, but different strains. Those authors found two bands, an expected one of 300 bp and an unexpected fragment of 400 bp, and that result was reproduced in this study by PCR amplification of E. faecium CRL 39.…”

Section: Discussioncontrasting

confidence: 99%

“…Considering the contrasting sizes and intensities obtained, it was not clear whether the fragments found in our study corresponded to amplified rDNA, since the yield of the 200 bp fragment was significantly smaller than the ones amplified by [17]. Thus, only the 200 bp product obtained from amplification of CRL 183 and the 300 bp product from CRL 39 were partially sequenced.…”

Section: Discussionmentioning

confidence: 73%

“…Those authors found two bands, an expected one of 300 bp and an unexpected fragment of 400 bp, and that result was reproduced in this study by PCR amplification of E. faecium CRL 39. The production of more than one band may be explained by differences in the region of insertion of the gene in the chromosome [17].…”

Section: Discussionmentioning

confidence: 99%

“…DNA fragments encoding the 16S-23S rRNA spacer region were amplified, using the primer combination Enf 1 (5'-AT-TACGGAGACTACACACTTTG-3') and Ent 2 (5'-TAGCCATAGAAGTTACAT-CAAG-3'), as detailed in [17].…”

Section: S Rdna Amplificationmentioning

confidence: 99%

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Demonstration of the cellular viability and safety ofEnterococcus faeciumCRL 183 in long-term experiments

Cano

Valentini

Sivieri

et al. 2007

Lait

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-Enterococcus faecium CRL 183, a strain isolated from NSLAB cheese starter, has been the focus of much research on its potential probiotic capacity, although its survival through the gastrointestinal tract has not been demonstrated so far. In order to determine the capacity of E. faecium CRL 183 to survive such conditions, this strain was administered daily to rats for 30 weeks. The experimental animals were divided into Group I: those that did not receive E. faecium, Group II: those that received a pure culture of E. faecium CRL 183 and Group III: animals that received E. faecium CRL 183 in the form of a fermented soy-based product. Faecal samples were collected at the beginning and at the 50%, 75% and 100% stages of the experimental period. Isolation and counts of Enterococcus were carried out on KF selective media. To distinguish the various Enterococcus species in the faeces, biochemical (API Strep 20) and molecular (PCR) tests were performed. Initially, E. faecium was absent from the intestinal flora of the rats; however, after 15 weeks of administration, E. faecium could be recovered from the faeces of Groups II and III, demonstrating that E. faecium CRL 183 was able to survive gastrointestinal transit under the study conditions. This is further evidence of the probiotic qualities of this strain. The safety of the strain was also investigated with regard to body weight and serum biochemical analysis. Résumé -Démonstration de la viabilité cellulaire et de l'innocuité de Enterococcus faecium CRL 183 dans des expériences sur le long terme. Enterococcus faecium CRL 183, souche isolée à partir de bactéries lactiques non levain en transformation fromagère, a été l'objet de nombreuses recherches sur sa capacité à être un probiotique potentiel, bien que sa survie dans le tractus digestif ne soit pas encore bien démontrée à ce jour. Afin de déterminer la capacité de E. faecium CRL 183 à survivre dans de telles conditions, cette souche a été administrée quotidiennement à des rats pendant 30 semaines. Les animaux ont été divisés en 3 groupes expérimentaux : le groupe I ne recevant pas E. faecium, le groupe II recevant une culture pure de E. faecium CRL 183, et le groupe III recevant E. faecium CRL 183 sous la forme d'un produit fermenté à base de soja. Les échantillons de fécès ont été collectés au début et à 50 %, 75 % et 100 % de la période d'expérimentation. L'isolement et la numération des Enterococcus ont été réalisés sur milieu sélectif KF. Des tests biochimiques (API Strep 20) et moléculaires (PCR) ont été réalisés pour distinguer les espèces isolées des fécès. Initialement, E. faecium était absent de la flore intestinale des rats ; cependant, après 15 semaines d'administration, E. faecium a pu être retrouvé dans les fécès des groupes II et III, démontrant que E. faecium CRL 183 était capable de survivre dans les conditions du transit gastrointestinal. L'innocuité de la souche a également été étudiée au regard du poids corporel et des analyses biochimiques plasmatiques.probiotique / Enterococcus faeci...

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Section: Discussioncontrasting

confidence: 99%

Section: Discussionmentioning

confidence: 73%

Section: Discussionmentioning

confidence: 99%

Section: S Rdna Amplificationmentioning

confidence: 99%

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Demonstration of the cellular viability and safety ofEnterococcus faeciumCRL 183 in long-term experiments

Cano

Valentini

Sivieri

et al. 2007

Lait

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“…A 1 kb Ladder Plus (Invitrogen, Life Technologies, Carlsbad, CA, USA) was included as molecular size standard. Finally, the gels were stained with ethidium bromide and photographed under UV illumination [ 39 , 40 ].…”

Section: Methodsmentioning

confidence: 99%

Probiotic Soy Product Supplemented with Isoflavones Improves the Lipid Profile of Moderately Hypercholesterolemic Men: A Randomized Controlled Trial

et al. 2016

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Background: Cardiovascular disease is the leading cause of worldwide morbidity and mortality. Several studies have demonstrated that specific probiotics affect the host’s metabolism and may influence the cardiovascular disease risk. Objectives: The aim of this study was to investigate the influence of an isoflavone-supplemented soy product fermented with Enterococcus faecium CRL 183 and Lactobacillus helveticus 416 on cardiovascular risk markers in moderately hypercholesterolemic subjects. Design: Randomized placebo-controlled double-blind trial Setting: São Paulo State University in Araraquara, SP, Brazil. Participants: 49 male healthy men with total cholesterol (TC) >5.17 mmol/L and <6.21 mmol/L Intervention: The volunteers have consumed 200 mL of the probiotic soy product (group SP-1010 CFU/day), isoflavone-supplemented probiotic soy product (group ISP–probiotic plus 50 mg of total isoflavones/100 g) or unfermented soy product (group USP-placebo) for 42 days in a randomized, double-blind study. Main outcome measures: Lipid profile and additional cardiovascular biomarkers were analyzed on days 0, 30 and 42. Urine samples (24 h) were collected at baseline and at the end of the experiment so as to determine the isoflavones profile. Results: After 42 days, the ISP consumption led to improved total cholesterol, non-HDL-C (LDL + IDL + VLDL cholesterol fractions) and electronegative LDL concentrations (reduction of 13.8%, 14.7% and 24.2%, respectively, p < 0.05). The ISP and SP have prevented the reduction of HDL-C level after 42 days. The C-reactive protein and fibrinogen levels were not improved. The equol production by the ISP group subjects was inversely correlated with electronegative LDL concentration. Conclusions: The results suggest that a regular consumption of this probiotic soy product, supplemented with isoflavones, could contribute to reducing the risk of cardiovascular diseases in moderately hypercholesterolemic men, through the an improvement in lipid profile and antioxidant properties.

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Einsatz molekularer Methoden für Starterkulturen

Ehrmann

Pavlovic²

2010

Molekularbiologische Methoden in Der Lebensmittelanalytik

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Differentiation of Enterococcus faecium from Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus strains by PCR and dot-blot hybridisation

Cited by 7 publications

References 7 publications

Demonstration of the cellular viability and safety ofEnterococcus faeciumCRL 183 in long-term experiments

Demonstration of the cellular viability and safety ofEnterococcus faeciumCRL 183 in long-term experiments

Probiotic Soy Product Supplemented with Isoflavones Improves the Lipid Profile of Moderately Hypercholesterolemic Men: A Randomized Controlled Trial

Einsatz molekularer Methoden für Starterkulturen

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