Enterococcus haemoperoxidus sp. nov. and Enterococcus moraviensis sp. nov., isolated from water.

Švec, Pavel; Devriese, Luc; Sedláček, Ivo; Baele, Margo; Vancanneyt, Marc; Haesebrouck, Freddy; Swings, Jean; Doškař, Jiřı́

doi:10.1099/00207713-51-4-1567

Cited by 68 publications

(24 citation statements)

References 31 publications

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“…Indeed, novel enterococcal species have been identified in recent years from water (38)(39)(40) and fecal (41,42) samples. While the relative abundances of novel enterococcal species in water samples are unknown, these results indicate that there is a need for further investigation of enterococcal diversity in both fecal and environmental samples.…”

Section: Resultsmentioning

confidence: 99%

Development of Quantitative PCR Assays Targeting the 16S rRNA Genes of Enterococcus spp. and Their Application to the Identification of Enterococcus Species in Environmental Samples

Ryu

Henson

Elk

et al. 2013

Appl Environ Microbiol

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eThe detection of environmental enterococci has been determined primarily by using culture-based techniques that might exclude some enterococcal species as well as those that are nonculturable. To address this, the relative abundances of enterococci were examined by challenging fecal and water samples against a currently available genus-specific assay (Entero1). To determine the diversity of enterococcal species, 16S rRNA gene-based group-specific quantitative PCR (qPCR) assays were developed and evaluated against eight of the most common environmental enterococcal species. Partial 16S rRNA gene sequences of 439 presumptive environmental enterococcal strains were analyzed to study further the diversity of enterococci and to confirm the specificities of group-specific assays. The group-specific qPCR assays showed relatively high amplification rates with targeted species (>98%), although some assays cross-amplified with nontargeted species (1.3 to 6.5%). The results with the group-specific assays also showed that different enterococcal species co-occurred in most fecal samples. The most abundant enterococci in water and fecal samples were Enterococcus faecalis and Enterococcus faecium, although we identified more water isolates as Enterococcus casseliflavus than as any of the other species. The prevalence of the Entero1 marker was in agreement with the combined number of positive signals determined by the group-specific assays in most fecal samples, except in gull feces. On the other hand, the number of group-specific assay signals was lower in all water samples tested, suggesting that other enterococcal species are present in these samples. While the results highlight the value of genus-and group-specific assays for detecting the major enterococcal groups in environmental water samples, additional studies are needed to determine further the diversity, distributions, and relative abundances of all enterococcal species found in water.

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Section: Resultsmentioning

confidence: 99%

Development of Quantitative PCR Assays Targeting the 16S rRNA Genes of Enterococcus spp. and Their Application to the Identification of Enterococcus Species in Environmental Samples

Ryu

Henson

Elk

et al. 2013

Appl Environ Microbiol

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“…Some isolates were clearly identified as either E. casseliflavus or E. flavescens, while other isolates were positive for both sets of primers. In addition, the species E. haemoperoxidus and E. moraviensis were not included in the multiplex PCR because sodA sequences from multiple isolates were not available for comparison (20). Moreover, although E. seriolicida and E. solitarius may eventually be reclassified into other genera, they are presently classified as enterococci and were included in the analysis (21,23).…”

Section: Discussionmentioning

confidence: 99%

Use of a Genus- and Species-Specific Multiplex PCR for Identification of Enterococci

2004

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The 16S rRNA gene has previously been used to develop genus-specific PCR primers for identification of enterococci. In addition, the superoxide dismutase gene (sodA) has been identified as a potential target for species differentiation of enterococci. In this study, Enterococcus genus-specific primers developed by Deasy et al. (E1/E2) were incorporated with species-specific primers based upon the superoxide dismutase (sodA) gene for development of a multiplex PCR. This assay provides simultaneous genus and species identification of 23 species of enterococci using seven different reaction mixtures. Accuracy of identification of the multiplex PCR was determined by comparisons to standard biochemical testing, the BBL Crystal kit, VITEK, and API Rapid ID 32 Strep. Isolates from swine feces, poultry carcasses, environmental sources, and retail food were evaluated and, overall, results for 90% of the isolates tested by PCR agreed with results obtained using standard biochemical testing and VITEK. Eighty-five percent and 82% of PCR results agreed with results from the API Rapid ID 32 Strep and BBL Crystal tests, respectively. With the exception of concurrence between identification using standard biochemical testing and VITEK (85%) and between BBL Crystal and VITEK (83%), the percent agreement for PCR was higher than or equal to any other pairwise comparison. Multiplex PCR for genus and species determination of enterococci provides an improved, rapid method for identification of this group of bacteria.

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“…Eleven isolates, obtained from Lisbon Veterinarian Faculty, were reported as dog-infecting strains. When phenotypic and molecular methods (6,33,35) The species E. villorum (45), E. haemoperoxidus and E. moraviensis (40), E. porcinus and E. ratti (43), and E. pallens and E. gilvus (44) were not included in the study, since they have been proposed and accepted as new species more recently.…”

Section: Microorganismsmentioning

confidence: 99%

Comparative Study Using Type Strains and Clinical and Food Isolates To Examine Hemolytic Activity and Occurrence of the cyl Operon in Enterococci

et al. 2003

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The hemolytic ability, the presence of cyl genes, and the diagnostic accuracy of cytolysin molecular detection were investigated in the genus Enterococcus by using 164 strains from 20 different species (26 reference strains, 42 clinical isolates from human and veterinary origin, and 96 isolates from ewe cheese and milk). Hemolysis was assayed with sheep and horse erythrocytes and under aerobic or anaerobic conditions. Screening of cytolysin genes (cylL L , cylL S , cylM, cylB, and cylA) was performed with new specific primers and the anaerobic assay of beta-hemolysis was used as the "gold standard" for the evaluation of cyl gene-based PCRs. Since beta-hemolysis and cyl genes were found in 10 and 14 species, respectively, the hemolytic ability seems to be spread throughout the genus Enterococcus. Beta-hemolysis was observed in 6 of 26 (23%) reference strains, 14 of 42 (33%) clinical isolates, and 6 of 96 (6%) food isolates. The presence of cyl genes was detected in 15 of 26 (58%) reference strains, 37 of 42 (88%) clinical isolates, and 67 of 96 (70%) food isolates. These data indicate a virulence potential in food isolates, reinforcing the need of their safety assessment. Analysis of phenotypicgenotypic congruence suggests a divergent sequence evolution of cyl genes and the effect of environmental factors in the regulation of cytolysin expression. Evaluation of the diagnostic accuracy of cytolysin molecular detection points to cylL L -based PCR and cylL L L S MBA-based PCR as the most reliable approaches. Nevertheless, the low sensitivity (46%) and gene variability indicated by our study strongly recommend the phenotypic assay for the assessment of hemolytic ability in enterococci, followed by the molecular screening of cyl genes in nonhemolytic strains to evaluate their virulence potential.

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Enterococcus haemoperoxidus sp. nov. and Enterococcus moraviensis sp. nov., isolated from water.

Cited by 68 publications

References 31 publications

Development of Quantitative PCR Assays Targeting the 16S rRNA Genes of Enterococcus spp. and Their Application to the Identification of Enterococcus Species in Environmental Samples

Development of Quantitative PCR Assays Targeting the 16S rRNA Genes of Enterococcus spp. and Their Application to the Identification of Enterococcus Species in Environmental Samples

Use of a Genus- and Species-Specific Multiplex PCR for Identification of Enterococci

Comparative Study Using Type Strains and Clinical and Food Isolates To Examine Hemolytic Activity and Occurrence of the cyl Operon in Enterococci

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