In situ Identification of Lactococci, Enterococci and Streptococci

Beimfohr, Claudia; Krause, Angela; Amann, Rudolf; Ludwig, Wolfgang; Schleifer, Karl‐Heinz

doi:10.1016/s0723-2020(11)80279-1

Cited by 123 publications

(81 citation statements)

References 21 publications

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“…After drying for 10 min, slides were dehydrated in 50, 80 and 100% ethanol each for 3 min. To optimize test conditions for the Lba probe, samples were incubated with 20 ml of lysozyme (10 mg ml À1 ) at 41C for 10 min as described previously (Beimfohr et al, 1993). Hybridization was carried out as described by Manz et al (1992).…”

Section: Yoghurt and Dietmentioning

confidence: 99%

Lactobacillus acidophilus 74-2 and Bifidobacterium animalis subsp lactis DGCC 420 modulate unspecific cellular immune response in healthy adults

Friedrich²,

et al. 2007

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Objective: It was determined whether a combination of Lactobacillus acidophilus (L. acidophilus) 74-2 and Bifidobacterium animalis subsp lactis DGCC 420 (B. lactis 420) affect the faecal microbiota as well as immunological parameters and blood lipids in healthy adults. Design: A placebo-controlled, double-blinded, randomized crossover trial was conducted. Subjects: Twenty-six healthy volunteers (mean age 25 years) were recruited by advertising in academical buildings. All of them completed the study. Methods: After 3-week run-in period, half of the volunteers consumed 300 g/day of yoghurt supplement containing probiotic strains L. acidophilus 74-2 and B. lactis 420, and the other half received the placebo product for a period of 5 weeks. The two groups were crossed during the following 5-week period. Blood and faecal samples were collected at the end of each period. The faecal content of probiotic bacteria, faecal short-chain fatty acids (SCFA), serum lipids and plasma immune system biomarkers were evaluated. Results: Faecal proportions of L. acidophilus and of B. lactis increased significantly from 0.02 to 0.19 and 0.4 to 1.4% (Po0.05), respectively. Percentages of granulocytes and monocytes showing phagocytic activity were significantly elevated from 92 to 95% during probiotic intervention, whereas their oxidative burst activity and specific immune parameters remained unaffected. Fecal SCFA and serum cholesterol levels were not influenced by the probiotics. However, serum concentrations of triacylglyceroles decreased significantly by 11.6% (Po0.05) in the probiotic supplementation period. Conclusions: L. acidophilus and B. lactis were recovered in faeces in significantly elevated numbers after supplementation. They are able to modulate unspecific cellular immune response indicated by the increased phagocytic activity.

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Section: Yoghurt and Dietmentioning

confidence: 99%

Lactobacillus acidophilus 74-2 and Bifidobacterium animalis subsp lactis DGCC 420 modulate unspecific cellular immune response in healthy adults

Friedrich²,

et al. 2007

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“…Fluorescent whole cell hybridization of BB-12-like colonies Whole cell hybridization was performed as published previously (Beimfohr et al, 1993) with some modifications. The fixation step was omitted and instead the cell material from purified isolates from the semiselective media was directly smeared onto microscope slides (Marienfeld) and dried at 371C for 30 min.…”

Section: Spiking Of Crl-431 In Fecesmentioning

confidence: 99%

Dose–response study of probiotic bacteria Bifidobacterium animalis subsp lactis BB-12 and Lactobacillus paracasei subsp paracasei CRL-341 in healthy young adults

Larsen

Nielsen²,

Kæstel³

et al. 2006

Eur J Clin Nutr

109

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Objective: This study was performed to investigate the dose-response effects of supplementation with Bifidobacterium animalis subsp lactis (BB-12) and Lactobacillus paracasei subsp paracasei (CRL-431) on blood lipids, recovery from feces and bowel habits. Changes of the fecal microflora was analyzed in the 10 10 CFU/day probiotic and placebo group. Design: The study was designed as a randomized, placebo-controlled, double-blinded, parallel dose-response study. Subjects: Healthy young adults (18-40 years) were recruited by advertising in local newspapers. Of the 75 persons enrolled, 71 (46 women, 25 men, mean age 25.6 years (range 18-40 years)) completed the study. Intervention: The volunteers were randomly assigned into five groups receiving either placebo or a mixture of the two probiotics in the concentration of 10 8 , 10 9 , 10 10 or 10 11 CFU/day in 2 weeks run-in period, 3 weeks intervention and 2 weeks wash-out. Diary reporting bowel habits and well being (abdominal bloating, flatulence and headache) was kept for all 7 weeks and blood lipids, fecal recovery of BB-12 and CRL-431, as well as fecal microflora was tested before, immediately and 2 weeks after intervention. Results: The fecal recovery of BB-12 increased significantly (Po0.001) with increasing dose. In the group receiving 10 11 CFU/ day BB-12 was recovered from 13 out of 15 volunteers. CRL-431 was not recovered in any of the fecal samples. Supplementation with probiotics did not change the fecal bacterial composition. A significant linear increase in fecal consistency (looser stool) with increasing probiotic dose (P ¼ 0.018) was observed. No overall dose-response effect was found on the blood lipids. High doses of probiotics were well tolerated. Conclusion: A dose-related recovery of BB-12 from feces was observed. Sponsorship: The study was sponsored by Chr. Hansen A/S, Hoersholm, Denmark. Keywords: dose-response study; probiotics; cholesterol; recovery from feces; constipation European Journal of Clinical Nutrition IntroductionIt is generally accepted that the composition of the intestinal microflora influences the health and well being of humans. With the many publications showing beneficial effects of probiotic bacteria, there has been an increasing interest in the mechanism behind. Most clinical studies have tested only one dose of probiotics, ranging from 4 Â 10 8 CFU/day for determination of gastrointestinal colonization and immune modulation (Valeur et al., 2004) Contributors: DCE, CNL and KFM wrote the protocol. DCE performed the study. CNL did the microbiological analysis of fecal samples and study product. EB performed the fluorescent whole cell hybridization of BB-12 like colonies. MB performed the PFGE of CRL-431 like colonies. PK performed the statistical analysis. CNL and SN wrote the first draft of the paper and all contributors participated in the revision and final approval of the paper.

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“…Current methods of identification are based on phenotypic characteristics that may show variation on testing, and for which the tests are difficult to perform and often take up to several days for a final result (Facklam & Collins, 1989). In recent years, authors have described molecular methods for the detection and identification of Enterococcus species by the use of labelled oligonucleotide probes based on 16S and 23S rRNA genes (Behr et al, 2000;Beimfohr et al, 1993;Betzl et al, 1990;Monstein et al, 1998;Pryce et al, 1999). However, most of these studies use direct DNAhybridization techniques for the identification of enterococci.…”

Section: Introductionmentioning

confidence: 99%

“…However, most of these studies use direct DNAhybridization techniques for the identification of enterococci. Only a few studies describe whole cell in situ hybridization for identification and enumeration of enterococci (Beimfohr et al, 1993;Harmsen et al, 1999).…”

Section: Introductionmentioning

confidence: 99%

“…The studies using FISH for detection of enterococci that have been published so far have been hampered by the fact that the specificities of the probes were not tested in different mixed bacterial populations or that they were only focused on one specific type of material (Beimfohr et al, 1993;Harmsen et al, 1999;Jansen et al, 2000). In this study, we describe the design of two fluorescent-labelled oligonucleotides that specifically detect the 16S rRNA of E. faecalis or the combination of E. faecium, Enterococcus hirae, Enterococcus mundtii, Enterococcus villorum and Enterococcus saccharolyticus.…”

Section: Introductionmentioning

confidence: 99%

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Fluorescent in situ hybridization with specific DNA probes offers adequate detection of Enterococcus faecalis and Enterococcus faecium in clinical samples

Waar

Degener

Luyn

et al. 2005

Journal of Medical Microbiology

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Enterococcus faecalis and Enterococcus faecium are among the leading causes of hospitalacquired infections. Reliable and quick identification of E. faecalis and E. faecium is important for accurate treatment and understanding their role in the pathogenesis of infections. Fluorescent in situ hybridization (FISH) of whole bacterial cells with oligonucleotides targeted at the 16S rRNA molecule leads to a reduced time to identification. In clinical practice, FISH therefore can be used in situations in which quick identification is necessary for optimal treatment of the patient. Furthermore, the abundance, spatial distribution and bacterial cell morphology can be observed in situ. This report describes the design of two fluorescent-labelled oligonucleotides that, respectively, detect the 16S rRNA of E. faecalis and the 16S rRNA of E. faecium, Enterococcus hirae, Enterococcus mundtii, Enterococcus villorum and Enterococcus saccharolyticus. Different protocols for the application of these oligonucleotides with FISH in different clinical samples such as faeces or blood cultures are given. Enterococci in a biofilm attached to a biomaterial were also visualized. Embedding of the biomaterial preserved the morphology and therefore the architecture of the biofilm could be observed. The usefulness of other studies describing FISH for detection of enterococci is generally hampered by the fact that they have only focused on one material and one protocol to detect the enterococci. However, the results of this study show that the probes can be used both in the routine laboratory to detect and determine the enterococcal species in different clinical samples and in a research setting to enumerate and detect the enterococci in their physical environment.

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In situ Identification of Lactococci, Enterococci and Streptococci

Cited by 123 publications

References 21 publications

Lactobacillus acidophilus 74-2 and Bifidobacterium animalis subsp lactis DGCC 420 modulate unspecific cellular immune response in healthy adults

Lactobacillus acidophilus 74-2 and Bifidobacterium animalis subsp lactis DGCC 420 modulate unspecific cellular immune response in healthy adults

Dose–response study of probiotic bacteria Bifidobacterium animalis subsp lactis BB-12 and Lactobacillus paracasei subsp paracasei CRL-341 in healthy young adults

Fluorescent in situ hybridization with specific DNA probes offers adequate detection of Enterococcus faecalis and Enterococcus faecium in clinical samples

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