Lactococcus garvieae endocarditis: identification by 16S rRNA and sodA sequence analysis.
ABSTRACT Lactococcus garvieae is only rarely isolated from clinical specimens. We report a case of prosthetic valve endocarditis caused by L. garvieae in an elderly patient. Molecular methods based on the 16S rRNA and sodA(int) gene sequences confirmed the phenotypic identification of this opportunistic human pathogen.
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ABSTRACT: The 16S rRNA sequences of enterococcal species E. faecium, E. faecalis, E. gallinarum, E. casseliflavus/flavescens, E. dispar, E. pseudoavium, E. sulfureus, E. malodoratus, E. raffinosus, E. cecorum, E. hirae, E. saccharolyticus, E. seriolicida, E. mundtii, E. avium, E. durans, E. columbae, and E. solitarius are presented herein. These data were utilized to confirm the species identification of two nonmotile E. gallinarum isolates which had been previously phenotypically identified as E. faecium. The implications of this finding are discussed.Journal of Clinical Microbiology 12/1998; 36(11):3399-407. · 4.07 Impact Factor
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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.Journal of Clinical Microbiology 01/2000; 38(1):415-8. · 4.07 Impact Factor
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ABSTRACT: Nucleic acid hybridization studies and immunological relationships of superoxide dismutase demonstrated that Streptococcus lactis (and its subspecies), Lactobacillus xylosus, Lactobacillus hordniae, S. garvieae, S. plantarum and S. raffinolactis are closely related to each other but not to other streptococci. Therefore we propose that these taxa be transferred to a new genus Lactococcus gen.nov. as Lactococcus lactis subsp. lactis (including former S. lactis subsp. diacetilactis and Lactobacillus xylosus) comb.nov., L. lactis subsp. cremoris comb.nov., L. lactis subsp. hordniae comb.nov., L. garvieae comb.nov., L. plantarum comb.nov. and L. raffinolactis comb.nov. The relatedness of these organisms has also been demonstrated by the similarity of their lipoteichoic acid structures, lipid pattern, fatty acid and menaquinone compositions. Motile ‘S. lactis’ strains are not genetically related to the lactococci although they possess the group N antigen. They also differ in their lipid composition from lactococci and may represent a new taxon. A description of the genus Lactococcus and emended descriptions of the corresponding species are given. The main characteristics for the differentiation of lactococci are listed in Table 8.Systematic and Applied Microbiology 09/1985; 6(2):183–195. · 3.29 Impact Factor
Lactococcus garvieae endocarditis: identification
by 16S rRNA and sodA sequence analysis
Vincent Fihmana,b,*, Laurent Raskineb, Zina Barrouc, Claire Kiffelc,
Jacques Riahib, Be ´atrice Berc ¸ota,b, Marie-Jose ´ Sanson-Le Porsa,b
aFaculte ´ de Me ´decine Lariboisie `re-Saint Louis, Universite ´ Paris 7, Paris, France
bService de Bacte ´riologie-Virologie, Groupe Hospitalier Lariboisie `re-Fernand Widal,
Assistance Publique-Ho ˆpitaux de Paris, Paris, France
cService de Ge ´rontologie, GroupeHospitalier Lariboisie `re-Fernand Widal, Assistance Publique-Ho ˆpitauxde
Paris, Paris, France
Accepted 22 April 2005
Available online 3 June 2005
16S rRNA gene;
We report a case of prosthetic valve endocarditis caused by L. garvieae in an elderly
patient. Molecular methods based on the 16S rRNA and sodAintgene sequences
confirmed the phenotypic identification of this opportunistic human pathogen.
Q 2005 The British Infection Society. Published by Elsevier Ltd. All rights reserved.
Lactococcus garvieae is only rarely isolated from clinical specimens.
An 86-year-old woman was admitted to our hospital
in June 2004 because of right hip pain for 3 weeks
and recent onset fever. She had undergone aortic
valve replacement with a bioprosthesis 6 years
previously and cholecystectomy for small bowel
obstruction complicated by a hernia 8 years
previously. She had been breathlessness during
exercise since November 2003 and had a duodenal
ulcer. She had been given ceftriaxone (1 g once a
day for 7 days) 1 month earlier for putative
pneumonia. Upon admission she was receiving
acebutolol and omeprazole.
Physical examination revealed respiratory dis-
tress with blood pressure of 144/87 mmHg, heart
rate 79 beats/min, respiratory rate 20 breaths/min
and an oral temperature of 38.1 8C. No evidence of
cardiac murmur or crackle was noted. The labora-
tory findings upon admission were: hemoglobin
level of 9.6 g/dl with mean corpuscular volume of
84 fl, white blood cell count of 15!109/l with 81%
neutrophils, platelet count of 487!109/l. The
C-reactive protein concentration was 94 mg/l.
A chest X-ray was normal except for cardiomegaly.
The patient was treated with intravenous
After 12 h of incubation, the three sets of blood
cultures all became positive. Direct examination
revealed short chains of Gram-positive cocci. This
facultative anaerobic, non-hemolytic, catalase-
negative bacterium formed small colonies on
blood agar. The API rapid ID 32 Strep system
Journal of Infection (2006) 52, e3–e6
0163-4453/$30.00 Q 2005 The British Infection Society. Published by Elsevier Ltd. All rights reserved.
*Corresponding author. Service de Bacte ´riologie-Virologie,
Ho ˆpital Lariboisie `re, 2 rue Ambroise Pare ´, 75475 Paris Cedex
10, France. Tel.: C33 1 49 95 65 44; fax: C33 1 49 95 85 37.
E-mail address: firstname.lastname@example.org (V. Fihman).
(bio-Me ´rieux, Marcy l’Etoile, France) was used to
identify the bacterium to species level as Lacto-
coccus garvieae with a 98.0% probability (profile
obtained 30333101130). Additional phenotypic
results were: Lancefield serogroup D-negative,
weak growth in broth containing 6.5% NaCl, growth
at 10 8C but not at 44 8C, positive bile-esculin
reaction, acid formation in mannitol broth but not
in sorbitol broth. The strain was confirmed to be
L. garvieae by genetic methods based on the 16S
rRNA1and sodAint2gene sequences. These two
genes were, respectively, 99.7 and 94.0% identical
to the corresponding sequences of the type strain of
L. garvieae ATCC 49156 (accession no. AF061005
and AJ387923). The E-test method was used to
determine MICs: penicillin G, 0.75; amoxicillin, 0.5;
cefotaxime, 0.38; vancomycin, 1.5; and teicopla-
nin, 0.38 mg/ml.
showed a mobile posterior aortic vegetation of
10 mm without bioprosthesis dehiscence or peri-
valvular leakage. Bone scintigraphy ruled out the
diagnosis of associated hip arthritis and spondylo-
discitis. Panoramic dentition radiography and colo-
noscopy did not show any source of infection.
Intravenous amoxicillin (150 mg/kg/day) and
gentamicin (2 mg/kg/day in a single dose) were
administered immediately after direct examination
of blood culture. After 4 weeks this treatment was
replaced by oral amoxicillin alone for 3 weeks. The
patient’s clinical status improved within 1 week.
A control TEE, performed after 4 weeks of optimal
dosing regimen as assessed by antibiotic dosage,
showed that the aortic vegetation had disappeared.
The antimicrobial treatment was stopped after
normalization of the C-reactive protein concen-
tration. The patient remains clinically well after
The Lactococcus genus was recently separated from
the Streptococcus genus on the basis of genetic
analysis including DNA–DNA relatedness and 16S
rRNA sequencing data.3The Lactococcus genus
comprises eight species and subspecies,4all of
which are facultative anaerobic, catalase-negative,
Gram-positive cocci that primarily produce lactic
acid from the fermentation of carbohydrates.5
These bacteria are very common in the environ-
ment and, until recently, were considered as
unusual pathogens in humans. According to a
MEDLINE search, subspecies of Lactococcus lactis,
used in fermented dairy products, have already
been incriminated in two cases of endocarditis.6,7
L. garvieae, which is another name for Enterococ-
cus seriolicida, is responsible for bovine mastitis
and septicaemia in several fish species.8It has also
been described as a human pathogen: four cases of
definite endocarditis have been reported, three
involving prosthetic valves and one involving a
native valve,9as well as one case of osteomyelitis10
and one case of bacteraemia associated with a liver
Our case is the fifth case of L. garvieae-related
endocarditis and the seventh case of Lactococcus
spp.-related endocarditis. Although Lactococcus
spp. are ubiquitous in food and in the environment,
their incidence in human endocarditis is currently
tolerant, Gram-positive cocci arranged in chains and pairsa
Differences between Lactococcus spp. involved in human infection and other catalase-negative, NaCl
Bacterium BEPYRLAPGrowth atMotility MAN SBLARAVancomycin
10 8C45 8C
L. lactis subsp. lactis
L. lactis subsp. cremoris
aBE, esculin hydrolysis in presence of 40% bile salts, PYR, pyrrolidonyl-b-naphthylamide hydrolysis; LAP, leucine-b-naphthylamide
hydrolysis; MAN, mannitol; SBL, sorbitol; ARA, arabinose.
V. Fihman et al. e4
very low compared with that of other lactic acid
bacteria like Enterococcus spp. and Lactobacillus
spp., which account for 5–15 and 0.1% of bacter-
aemia cases, respectively.12No virulence studies
have been conducted on lactococci to explain this
difference. Members of this genus are not abundant
in the normal human flora and seem to behave like
opportunistic pathogens in the elderly, immunode-
ficient subjects and individuals with prosthetic
valves. The patient described here presented two
of these three risk factors. Two negative urine
cultures and panoramic dentition radiography
failed to identify the source of infection. The
digestive tract was suspected based on a decrease
in gastric acidity induced by omeprazole, but
colonoscopy revealed only uncomplicated colon
diverticulosis and the patient did not regularly eat
fish or cheese.
The low prevalence of human lactococci infec-
tions may also be due to the difficulties involved in
identifying this bacterial genus. Indeed, the phe-
notypic characteristics of Lactococcus species are
similar to those of other genera, particularly
Enterococcus spp., which can result in misidentifi-
cation (Table 1). Our clinical isolate did not grow in
broth incubated at 44 8C for 72 h. This is a
characteristic of members of the Lactococcus
genus whereas enterococci grow readily in these
conditions in 24 h.5Furthermore, our isolate only
grew weakly in 6.5% NaCl broth whereas entero-
cocci usually grow well at this concentration.
Finally, this isolate had no zone of inhibition around
the clindamycin disk (MICO8 mg/ml) as already
described for L. garvieae strains but not for
L. lactis strains.13
Phenotypic identification is time-consuming and
may lead to misidentification. We demonstrate in
this report that molecular methods are a rapid
alternative. DNA extraction and 16S rRNA or sodA
gene sequencing can be performed within 48 h,
leading to correct identification (Fig. 1).
In conclusion, although not frequently recovered
from human clinical specimens, L. garvieae can be
considered as an authentic cause of endocarditis.
Proper phenotypic or molecular identification
should make it possible to correct the probable
1. Patel R, Piper KE, Rouse MS, Steckelberg JM, Uhl JR,
Kohner P, et al. Determination of 16S rRNA sequences of
enterococci and application to species identification of
nonmotile Enterococcus gallinarum isolates. J Clin Microbiol
2. Poyart C, Quesnes G, Trieu-Cuot P. Sequencing the gene
encoding manganese-dependent superoxide dismutase for
rapid species identification of enterococci. J Clin Microbiol
3. Schleifer KH, Kraus J, Dvorak C, Klipper-Balz R, Collins MD,
Fischer W, et al. Transfer of Streptococcus lactis and related
streptococci to the genus Lactococcus gen. nov. Syst Appl
4. Boone DR, Castenholz RW, Garrity GM. Bergey’s manual of
systematic bacteriology. 2nd ed. New York: Springer; 2001.
5. Facklam R, Elliott JA. Identification, classification, and
clinicalrelevance of catalase-negative,
cocci, excluding the streptococci and enterococci. Clin
Microbiol Rev 1995;8:479–95.
6. Mannion PT, Rothburn MM. Diagnosis of bacterial endocardi-
tis caused by Streptococcus lactis and assisted by immuno-
blotting of serum antibodies. J Infect 1990;21:317–8.
7. Wood HF, Jacobs K, McCarty M. Streptococcus lactis isolated
from a patient with subacute bacterial endocarditis. Am
J Med 1985;18:345–7.
8. Vela AI, Vazquez J, Gibello A, Blanco MM, Moreno MA,
Liebana P, et al. Phenotypic and genetic characterization of
Lactococcus garvieae isolated in Spain from lactococcosis
outbreaks and comparison with isolates of other countries
and sources. J Clin Microbiol 2000;38:3791–5.
catalase-negative, Gram-positive cocci. The trees were constructed by the neighbour-joining method.
Distance matrix tree showing relationships between 16S rRNA (A) and sodA (B) sequences from various
Lactococcus garvieae endocarditis e5
9. Fefer JJ, Ratzan KR, Sharp SE, Saiz E. Lactococcus garvieae
endocarditis: report of a case and review of the lature. Diagn
Microbiol Infect Dis 1998;32:127–30.
10. James PR, Hardman SM, Patterson DL. Osteomyelitis and
possible endocarditis secondary to Lactococcus garvieae: a
first case report. Postgrad Med J 2000;76:301–3.
11. Mofredj A, Baraka D, Kloeti G, Dumont JL. Lactococcus
garvieae septicaemia with liver abscess in an immunosup-
pressed patient. Am J Med 2000;109:513–4.
12. Salminen S, von Wright A, Morelli L, Marteau P, Brassart D,
de Vos WM, et al. Demonstration of safety of probiotics—a
review. Int J Food Microbiol 1998;44:93–106.
13. Elliott JA, Facklam RR. Antimicrobial susceptibilities of
Lactococcus lactis and Lactococcus garvieae and a proposed
method to discriminate between them. J Clin Microbiol
V. Fihman et al.e6