scholarly journals Identification of Enterococcus Species and Phenotypically Similar Lactococcus andVagococcus Species by Reverse Checkerboard Hybridization to Chaperonin 60 Gene Sequences

2000 ◽  
Vol 38 (11) ◽  
pp. 3953-3959 ◽  
Author(s):  
Swee Han Goh ◽  
Richard R. Facklam ◽  
Michelle Chang ◽  
Janet E. Hill ◽  
Gregory J. Tyrrell ◽  
...  
Keyword(s):  
Lactococcus Lactis ◽  
Single Pair ◽  
Degenerate Primers ◽  
Lactococcus Garvieae ◽  
Enterococcus Mundtii ◽  
Enterococcus Casseliflavus ◽  
Chaperonin 60 ◽  
Enterococcus Gallinarum ◽  
Method R ◽  
Enterococcus Cecorum

Data from four recent studies (S. H. Goh et al., J. Clin. Microbiol. 36:2164–2166, 1998; S. H. Goh et al., J. Clin. Microbiol. 34:818–823, 1996; S. H. Goh et al., J. Clin. Microbiol. 35:3116–3121, 1997; A. Y. C. Kwok et al., Int. J. Syst. Bacteriol. 49:1181–1192, 1999) suggest that an approximately 600-bp region of the chaperonin 60 (Cpn60) gene, amplified by PCR with a single pair of degenerate primers, has utility as a potentially universal target for bacterial identification (ID). This Cpn60 gene ID method correctly identified isolates representative of numerous staphylococcal species and Streptococcus iniae, a human and animal pathogen. We report herein that this method enabled us to distinguish clearly between 17 Enterococcus species (Enterococcus asini, Enterococcus rattus,Enterococcus dispar, Enterococcus gallinarum,Enterococcus hirae, Enterococcus durans,Enterococcus cecorum, Enterococcus faecalis,Enterococcus mundtii, Enterococcus casseliflavus, Enterococcus faecium,Enterococcus malodoratus, Enterococcus raffinosus, Enterococcus avium, Enterococcus pseudoavium, Enterococcus new sp. strain Facklam, andEnterococcus saccharolyticus), and Vagococcus fluvialis, Lactococcus lactis, and Lactococcus garvieae. From 123 blind-tested samples, only two discrepancies were observed between the Facklam and Collins phenotyping method (R. R. Facklam and M. D. Collins, J. Clin. Microbiol. 27:731–734, 1989) and the Cpn60 ID method. In each case, the discrepancies were resolved in favor of the Cpn60 ID method. The species distributions of the 123 blind-tested isolates wereEnterococcus new sp. strain Facklam (ATCC 700913), 3;E. asini, 1; E. rattus, 4; E. dispar, 2; E. gallinarum, 20; E. hirae, 9; E. durans, 9; E. faecalis, 12; E. mundtii, 3; E. casseliflavus, 8; E. faecium, 25; E. malodoratus, 3; E. raffinosus, 8; E. avium, 4; E. pseudoavium, 1; an unknown Enterococcus clinical isolate, sp. strain R871; Vagococcus fluvialis, 4;Lactococcus garvieae, 3; Lactococcus lactis, 3;Leuconostoc sp., 1; and Pediococcus sp., 1. The Cpn60 gene ID method, coupled with reverse checkerboard hybridization, is an effective method for the identification ofEnterococcus and related organisms.

scholarly journals Sequencing the Gene Encoding Manganese-Dependent Superoxide Dismutase for Rapid Species Identification of Enterococci

2000 ◽  
Vol 38 (1) ◽  
pp. 415-418
Author(s):  
Claire Poyart ◽  
Gilles Quesnes ◽  
Patrick Trieu-Cuot
Keyword(s):  
Superoxide Dismutase ◽  
Bacterial Species ◽  
Single Species ◽  
Rrna Gene ◽  
Target Sequence ◽  
Single Pair ◽  
Degenerate Primers ◽  
Gene Encoding ◽  
Enterococcus Casseliflavus ◽  
Enterococcus Gallinarum

ABSTRACT Simple PCR and sequencing assays that utilize a single pair of degenerate primers were used to characterize a 438-bp-long DNA fragment internal ( sodA int ) to the sodA gene encoding the manganese-dependent superoxide dismutase in 19 enterococcal type strains ( Enterococcus avium , Enterococcus casseliflavus , Enterococcus cecorum , Enterococcus columbae , Enterococcus dispar , Enterococcus durans , Enterococcus faecalis , Enterococcus faecium , Enterococcus flavescens , Enterococcus gallinarum , Enterococcus hirae , Enterococcus malodoratus , Enterococcus mundtii , Enterococcus pseudoavium , Enterococcus raffinosus , Enterococcus saccharolyticus , Enterococcus seriolicida , Enterococcus solitarius , and Enterococcus sulfureus ). Sequence analysis of the sodA int fragments enabled reliable identification of 18 enterococcal species, including E. casseliflavus-E. flavescens and E. gallinarum . The sodA int fragments of E. casseliflavus and E. flavescens were almost identical (99.5% sequence identity), which suggests that they should be associated in a single species. Our results confirm that the sodA gene constitutes a more discriminative target sequence than 16S rRNA gene in differentiating closely related bacterial species.

scholarly journals Effects of Different Test Conditions on MICs of Food Animal Growth-Promoting Antibacterial Agents for Enterococci

1998 ◽  
Vol 36 (7) ◽  
pp. 1907-1911 ◽  
Author(s):  
Patrick Butaye ◽  
Luc A. Devriese ◽  
Freddy Haesebrouck
Keyword(s):  
Aerobic Incubation ◽  
Mueller Hinton ◽  
Enterococcus Mundtii ◽  
Enterococcus Casseliflavus ◽  
Enterococcal Species ◽  
Animal Growth ◽  
Growth Promoting ◽  
Enterococcus Gallinarum ◽  
Susceptibility Tests ◽  
Free Media

The influence of the addition of sheep blood to Mueller-Hinton II agar and the effects of aerobic incubation with or without CO2 and of anaerobic incubation were tested with bacitracin, tylosin, avoparcin, virginiamycin, avilamycin, narasin, and flavomycin on enterococci. The antibacterial activity of bambermycin (Flavomycin) was strongly inhibited by the addition of blood, except with the species Enterococcus faecium, Enterococcus mundtii, Enterococcus hirae, Enterococcus casseliflavus, and Enterococcus gallinarum, which were not susceptible to this antibiotic on blood-free medium. With all other antimicrobials except avoparcin and tylosin, the presence of blood resulted in MIC increases of 1 to 3 log2 differences. Incubation in aerobic or anaerobic atmospheres enriched with CO2 lowered the susceptibility of enterococci to tylosin and increased their susceptibility to avilamycin, narasin, and avoparcin. This effect was most pronounced in tests on blood-free media. Results of susceptibility tests incubated under anaerobiosis and in a CO2-enriched atmosphere did not differ. For all enterococcal species, the preferred conditions for testing the susceptibility are Mueller-Hinton II medium supplemented with blood and incubation in a CO2-enriched atmosphere. However, when onlyE. faecium and Enterococcus faecalis are being tested, Mueller-Hinton II medium without blood incubated aerobically gives satisfactory results.

scholarly journals Use of Tests for Acidification of Methyl-α-d-Glucopyranoside and Susceptibility to Efrotomycin for Differentiation of Strains ofEnterococcus and Some Related Genera

1998 ◽  
Vol 36 (6) ◽  
pp. 1584-1587 ◽  
Author(s):  
Maria Da Glória S. Carvalho ◽  
Lúcia M. Teixeira ◽  
Richard R. Facklam
Keyword(s):  
Enterococcus Faecalis ◽  
Enterococcus Faecium ◽  
Clinical Microbiology ◽  
Clinical Microbiology Laboratory ◽  
Microbiology Laboratory ◽  
Lactococcus Garvieae ◽  
Negative Results ◽  
Enterococcus Casseliflavus ◽  
Enterococcus Gallinarum ◽  
Atypical Strains

A total of 107 Enterococcus strains, 10Vagococcus fluvialis strains, and 8 Lactococcus garvieae strains were tested for acidification of methyl-α-d-glucopyranoside (MGP) and susceptibility to 100-μg efrotomycin (EFRO) disks. All 26 strains ofEnterococcus casseliflavus, including 3 nonmotile and 2 nonpigmented strains, acidified MGP and were resistant to EFRO. All 22 strains of Enterococcus gallinarum, including 5 nonmotile strains, also acidified MGP and were resistant to EFRO. None of the 26 strains of Enterococcus faecium acidified MGP, and all were susceptible to EFRO. Although all 12 Enterococcus faecalisstrains were also negative in the MGP test, they were resistant to EFRO. Other enterococcal strains gave variable results. All 10 strains of V. fluvialis and all 8 strains of L. garvieae gave positive and negative results, respectively, in the MGP test and were, respectively, resistant and susceptible to EFRO. These results indicate that tests of the production of acid from MGP and susceptibility to EFRO can be used as adjunct tests in the identification of typical and atypical strains of enterococci in the clinical microbiology laboratory.

scholarly journals First isolation of Enterococcus strains in pig faeces in Turkey and determination of antibiotic susceptibilities

2013 ◽  
Vol 82 (3) ◽  
pp. 231-235
Author(s):  
Kemal Metiner ◽  
Mine Anğ Küçüker ◽  
Özden Büyükbaba Boral ◽  
Özdem Anğ
Keyword(s):  
Antimicrobial Resistance ◽  
High Rate ◽  
Diffusion Method ◽  
Marmara Region ◽  
Disk Diffusion Method ◽  
Faecal Samples ◽  
Enterococcus Casseliflavus ◽  
Enterococcus Gallinarum ◽  
Enterococcus Cecorum

In this trial,Enterococcusstrains were isolated from a total of 69 faecal samples obtained from 238 pigs (105 pigs < 6 months old and 133 > 6 months old) on three pig farms located in Istanbul and Tekirdag Provinces in the Marmara Region of Turkey in the summer season of 2003. Forty-seven of the isolates were determined asEnterococcus faecium(68%), 15 isolates asEnterococcus faecalis(21.7%), three isolates asEnterococcus gallinarum(4.3%) and one of each asEnterococcus hirae(1.4%),Enterococcus casseliflavus(1.4%),Enterococcus cecorum(1.4%)andEnterococcus sulfurens(1.4%). In addition, antimicrobial susceptibilites of isolates were assessed through the disk diffusion method. Among 47E. faeciumisolates, 44 were determined to be resistant to erythromycin, 38 to ciprofloxacine, and three isolates were resistant to vancomycin. AllE. faecalisisolates were resistant to erythromycin (100%) and four were resistant to vancomycin (27%). FiveE. faecium(11%) and fiveE. faecalisisolates (33%) were found to exhibit intermediate resistance to vancomycin. In this study, isolates obtained from pig faeces were determined to exhibit a high rate of antimicrobial resistance. This study is the first report on isolation and determination of antimicrobial resistance of Enterocci in Turkey.

On the Microbiological Profile of Traditional Portuguese Sourdough

1999 ◽  
Vol 62 (12) ◽  
pp. 1416-1429 ◽  
Author(s):  
J. MIGUEL ROCHA ◽  
F. XAVIER MALCATA
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Lactococcus Lactis ◽  
Dominant Species ◽  
Geographic Locations ◽  
Microbiological Profile ◽  
Enterococcus Casseliflavus ◽  
Lactobacillus Spp

Traditional manufacture of bread from maize has been noted to play important roles from both economic and social standpoints; however, enforcement of increasingly strict hygiene standards requires thorough knowledge of the adventitious microbiota of the departing dough. To this goal, sourdough as well as maize and rye flours from several geographic locations and in two different periods within the agricultural year were assayed for their microbiota in sequential steps of quantification and identification. More than 400 strains were isolated and taxonomic differentiation between them was via Biomerieux API galleries (375 of which were successfully identified) following preliminary biochemical and morphological screening. The dominant groups were yeasts and lactic acid bacteria (LAB). The most frequently isolated yeasts were Saccharomyces cerevisiae and Candida pelliculosa. The most frequently isolated LAB were (heterofermentative) Leuconostoc spp. and (homo-fermentative) Lactobacillus spp.; L. brevis, L. curvatus, and L. lactis ssp. lactis were the dominant species for the Lactobacillus genera; Lactococcus lactis ssp. lactis for lactococci; Enterococcus casseliflavus, E. durans, and E. faecium for enterococci; and Streptococcus constellantus and S. equinus for streptococci.

scholarly journals Perfil de susceptibilidade a antimicrobianos em amostras de cocos Gram-positivos, catalase negativos, isoladas de mastite subclínica bubalina

2003 ◽  
Vol 23 (2) ◽  
pp. 47-51 ◽  
Author(s):  
Maria C.E. Vianni ◽  
Norma S. Lázaro
Keyword(s):  
Rio De Janeiro ◽  
Clinical Laboratory ◽  
National Committee ◽  
Lactococcus Garvieae ◽  
Enterococcus Gallinarum

Estudou-se o perfil de susceptibilidade a antimicrobianos em cocos Gram-positivos catalase negativos (21 amostras de Lactococcus garvieae e 6 de Enterococcus gallinarum), isoladas do leite de fêmeas com mastite subclínica e pertencentes a uma população composta por seis rebanhos bubalinos localizados no Estado do Rio de Janeiro. O teste utilizado foi o da difusão de discos em agar Müller Hinton, segundo recomendações do National Committee for Clinical Laboratory Standards - NCCLS, tendo sido testados discos com ampicilina (10mg), cefalotina (30mg), cefotaxima (30mg), cefoxitina (30mg), cloranfenicol (30mg), eritromicina (15mg), gentamicina (10mg), nitrofurantoína (300mg), norfloxacina (10mg), penicilina (10 UI), tetraciclina (30mg) e vancomicina (30mg). Os resultados evidenciaram que em se tratando de Lactococcus garvieae, o antimicrobiano mais eficiente foi o nitrofurantoína com 85,71% de sensibilidade, seguido da cefotaxima (61,90%), vancomicina (52,38%), norfloxacina (47,62%) e cefalotina (47,62%). A maior resistência foi desenvolvida frente a penicilina e ampicilina, com 95,24% de resistênciapara os dois antimicrobianos testados. O perfil de susceptibilidade desenvolvido pelas amostras de Enterococcus gallinarum, mostrou baixa sensibilidade frente aos antimicrobianos testados, onde os maiores índices foram observados frente eritromicina e gentamicina, com 33,34% de sensibilidade para ambos; quanto à resistência desenvolvida, foi possível observar 100% de resistência com relação a vancomicina e tetraciclina, seguindo-se cloranfenicol, penicilina, ampicilina, cefoxitina, cefalotina, cefotaxima, norfloxacina e nitrofurantoína, todas evidenciando uma resistência de 83,33% das amostras testadas.

scholarly journals PCR-RFLP of 16S ribosomal DNA to confirm the identification of Enterococcus gallinarum and Enterococcus casseliflavus isolated from clinical and food samples

2010 ◽  
Vol 43 (1) ◽  
pp. 100-101 ◽  
Author(s):  
Aline Weber Medeiros ◽  
Pedro d'Azevedo ◽  
Rebeca Inhoque Pereira ◽  
Ana Paula Cassenego ◽  
Sueli Van Der Sand ◽  
...  
Keyword(s):  
Pcr Amplification ◽  
Food Samples ◽  
Correct Identification ◽  
Accurate Identification ◽  
16S Ribosomal Dna ◽  
Pcr Rflp ◽  
Enterococcus Casseliflavus ◽  
Dna Pattern ◽  
Enterococcus Gallinarum ◽  
Rdna Analysis

INTRODUCTION: This study aimed to confirm the identification of Enterococcus gallinarum and Enterococcus casseliflavus isolated from clinical and food samples by PCR-RFLP. METHODS: Fifty-two strains identified by conventional biochemical exams were submitted to PCR amplification and digested with HinfI. Only 20 (38.5%) of the 52 strains showed a DNA pattern expected for E. gallinarum and E. casseliflavus. RESULTS: Analysis of the results of this study showed that E. gallinarum and E. casseliflavus are occasionally erroneously identified and confirmed the potential application of 16S rDNA analysis for accurate identification of these species. CONCLUSIONS: A correct identification is important to distinguish between intrinsic and acquired vancomycin resistance.

scholarly journals A Virulent Phage Infecting Lactococcus garvieae, with Homology to Lactococcus lactis Phages

2015 ◽  
Vol 81 (24) ◽  
pp. 8358-8365 ◽  
Author(s):  
Giovanni Eraclio ◽  
Denise M. Tremblay ◽  
Alexia Lacelle-Côté ◽  
Simon J. Labrie ◽  
Maria Grazia Fortina ◽  
...  
Keyword(s):  
Lactococcus Lactis ◽  
Common Ancestor ◽  
Burst Size ◽  
Open Reading Frames ◽  
Content Type ◽  
Virulent Phage ◽  
Lactococcus Garvieae ◽  
Double Stranded Dna ◽  
Compost Soil ◽  
Reading Frames

ABSTRACTA new virulent phage belonging to theSiphoviridaefamily and able to infectLactococcus garvieaestrains was isolated from compost soil. Phage GE1 has a prolate capsid (56 by 38 nm) and a long noncontractile tail (123 nm). It had a burst size of 139 and a latent period of 31 min. Its host range was limited to only twoL. garvieaestrains out of 73 tested. Phage GE1 has a double-stranded DNA genome of 24,847 bp containing 48 predicted open reading frames (ORFs). Putative functions could be assigned to only 14 ORFs, and significant matches in public databases were found for only 17 ORFs, indicating that GE1 is a novel phage and its genome contains several new viral genes and encodes several new viral proteins. Of these 17 ORFs, 16 were homologous to deduced proteins of virulent phages infecting the dairy bacteriumLactococcus lactis, including previously characterized prolate-headed phages. Comparative genome analysis confirmed the relatedness ofL. garvieaephage GE1 toL. lactisphages c2 (22,172 bp) and Q54 (26,537 bp), although its genome organization was closer to that of phage c2. Phage GE1 did not infect any of the 58L. lactisstrains tested. This study suggests that phages infecting different lactococcal species may have a common ancestor.

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