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Diagnostic Microbiology and Infectious Disease
isolation from conjunctival swab in Italy
G.M. Giammancoa,*, V. Di Marcob, I. Priolob, A. Intrivicic, F. Grimontd, P.A.D. Grimontd
aDipartimento di Igiene e Microbiologia “G. D’Alessandro”, Universita` di Palermo, Palermo, Italy
bIstituto Zooprofilattico Sperimentale della Sicilia “A. Mirri”, Palermo, Italy
cAzienda Sanitaria Locale n.6, Palermo, Italy
dUnite´ Biodiversite´ des Bacte´ries Pathoge`nes Emergentes, Institut Pasteur, Paris, France
Received 29 April 2002; accepted 20 June 2002
Presented in part: Societa` Italiana di Microbiologia
(SIM) 29th Congress, Genoa, Italy, November 2001.
was isolated from conjunctival swabs of a farmer suffering from purulent conjunctivitis. This species has
only recently been reported in Switzerland and Germany to be exclusively isolated from ocular surfaces. This represents the first isolationof C. macginleyi
in Italy indicating that its circulation is not geographically limited. 2002 Elsevier Science Inc. All rights reserved.
1. Case report
colonies of Gramϩ pleomorphic rods were observed after48 h incubation on SBA. No growth was observed on
The species Corynebacterium macginleyi
has been re-
chocolate agar and MacConkey agar. Gram’s stain revealed
cently described by Riegel and coll. (Riegel et al., 1995) as
coryneform rods forming arrangements similar to Chinese
a result of comprehensive investigation of lipophilic coryne-
letters. The commercial API Coryne system (bioMe´rieux,
bacteria. Since the original description including three
Marcy l’Etoile, France) was used according to the manu-
strains isolated from eye specimens, only 25 clinical isolates
facturer’s instructions for identification of the isolate. The
of C. macginleyi
have been reported in the literature. All of
numerical code 5100305 was obtained after 24 h incubation
these clinical strains have been isolated from conjunctival
and used for species identification with API Coryne data-
swabs in Switzerland (Funke et al., 1998) and Germany
base 2.0 (Funke et al., 1997). API Coryne system unequivo-
cously identified the isolate as C. macginleyi
and the nu-
We isolated C. macginleyi
from a conjunctival swab of a
merical code obtained corresponded to the one more
65 year old farmer living in a rural area in Western Sicily
frequently encountered by Funke and coll. (Funke et al.,
and suffering from bilateral conjunctivitis. According to the
1998). Moreover, an identical identification code was ob-
physician in charge, the patient showed conjunctival hype-
tained with C. macginleyi
type strain (ATCC 104099T).
raemia with a whitish discharge, loss of visual acuity and
The Corynebacterium isolate was confirmed to be li-
corneal ulcers, but with no signs of intraocular infection.
pophilic by testing its growth on Tween 80-supplemented
The conjunctival sample was cultured on Columbia agar
SBA (Riegel et al., 1995). Our isolate grew well on both 0.1
plates (Becton Dickinson BBL, Cockeysville, Md.) supple-
mented with 5% sheep blood (SBA) and chocolate agar
Antimicrobial susceptibility patterns were determined by
(Becton Dickinson) at 37°C both in a 5% CO -enriched
the agar diffusion method. Our strain was susceptible to a
atmosphere and in ambient air and on MacConkey agar
large panel of antibiotics including: amoxicillin/clavulanate,
(Becton Dickinson) at 37°C in ambient air. Very small
clindamycin, erythromycin, gentamicin, penicillin G, chlor-amphenicol, rifampicin, tetracycline, vancomycin, ce-foperazone, enrofloxacyn, and kanamycin.
In order to determine the phylogenetic position of our
* Corresponding author. Tel.: ϩ39-0916553663; fax: ϩ39-091343896.E-mail address:
firstname.lastname@example.org (G.M. Giammanco).
isolate, 16S rRNA gene was amplified by polymerase chain
0732-8893/02/$ – see front matter 2002 Elsevier Science Inc. All rights reserved.
PII: S 0 7 3 2 - 8 8 9 3 ( 0 2 ) 0 0 4 3 8 - 8
G.M. Giammanco et al. / Diagnostic Microbiology and Infectious Disease 44 (2002) 205–207
Fig. 1. Phylogenetic dendrogram based on 16S rDNA sequences of reference strains of lipophilic species belonging to Corynebacterium
genus and relativeposition of the first Italian C. macginleyi
reaction and sequenced by GenomeExpress (Montreuil,
is reported to predominantly affect already
France). The sequence of our clinical isolate was compared
injured conjunctivas or superinfect during conjunctivitis
and aligned to published 16S rRNA sequences searched
caused by other bacterial micro-organisms or viruses
with NCBI taxonomy browser (National Center for Biotech-
(Funke et al., 1998; Joussen et al., 2000). No other bacteria
nology Information, National Library of Medicine, Be-
could be isolated from our sample but, unfortunately, it was
thesda, MD, USA; http://www.ncbi.nlm.nih.gov) and re-
not submitted for viral or Chlamydia
isolation, so that bac-
trieved from GenBank. The retrieved 16S rRNA sequences
terial superinfection of a viral or chlamydial etiology can
belonged to nine reference strains of six different species of
including C. macginleyi
The patient recovered uneventfully three weeks after
strain, and three strains of CDC Groups F-1 and G. The
sampling following Colbiocin (chloramphenicol ϩ colistin
EMBL/GenBank accession numbers for the 16S rRNA se-
ϩ tetracycline) antibiotic drops treatment. According to the
quences were as follows: C. accolens
literature, our isolate was susceptible to a large panel of
X80500; C. afermentans afermentans
antibiotics including chloramphenicol and tetracycline. De-
X82054; C. afermentans lipophilum
CIP 103500T, X82055;
layed recovery could be explained by the scarce compliance
NCTC 3224T, X84444; C. jeikeium
of our patient toward therapy. Other three members of the
U87816; C. jeikeium
ATCC 43217, U87815; C. jeikeium
same family showed similar symptoms but they were not
ATCC 43734, U87823; C.
CDC Group F-1 CDC G5911,
sampled because they had already started antibiotic treat-
CDC Group F-1 G4330, X81905; C.
ment. They all recovered after antibiotic drops treatment
Group G CDC G5840, X80498; C. urealyticum
but, in the absence of isolation, the occurrence of a C.
43042T, X81913; C. macginleyi
ATCC 104099T, X80499.
epidemic remains a mere speculation.
Phylogenetic analysis included sequence alignment, cal-
Our finding of a case of conjunctivitis associated to C.
culation of percent sequence similarity, construction of a
isolation in Italy confirms Joussen’s and coll.
phylogenetic tree, and assessment of the tree topology by
indication that the presence of this micro-organism is not
bootstrap analysis, which was performed by Clustal method
geographically limited (Joussen et al., 2000).
with Weighted residue weight table using DNAstar software(DNASTAR Inc., Madison, WI, USA). In the phylogenetictree (Fig. 1) our clinical isolate clustered together with C.
published sequence showing 98.7% sequencesimilarity (0.4 divergence). 16S rDNA sequence similarity
Thanks are due to G. Rubino for excellent technical
above 97% should be considered sufficient to consider two
strains as belonging to the same species (Stackebrandt &Goebel, 1994). The 16S rRNA gene sequence obtained forthe Italian C. macginleyi
isolate has been deposited in theGenBank sequence data-base and given accession no.
As confirmed by 16S rDNA sequence analysis API
Funke, G., Renaud, F. N. R., Freney, J., & Riegel, P. (1997). Multicenter
Coryne system is affordable in identifying C. macginleyi
evaluation of the updated and extended API (RAPID) Coryne database2.0. Journal of Clinical Microbiology, 35,
isolates. Growth on Tween 80-supplemented SBA can be
Funke, G., Pagano-Niederer, M., & Bernauer, W. (1998). Corynebacterium
used to detect lipophilic isolates but further biochemical
has to date been isolated exclusively from conjunctival
tests are needed for species identification.
swabs. Journal of Clinical Microbiology, 36,
G.M. Giammanco et al. / Diagnostic Microbiology and Infectious Disease 44 (2002) 205–207
Joussen, A. M., Funke, G., Joussen, F., & Herbertz, G. (2000). Coryne-
of Corynebacterium macginleyi
sp. nov. International Journal of Sys-
: a conjunctiva specific pathogen. British Journal
temic Bacteriology, 45,
of Ophthalmology, 84,
Stackebrandt, E., & Goebel, B. M. (1994). Taxonomic note: a place for
Riegel, P., Ruimy, R., de Briel, D., Prevost, G., Jehl, F., Christen, R., &
DNA-DNA reassociation and 16S rRNA sequence analysis in the
Monteil, H. (1995). Genomic diversity and phylogenetic relationships
present species definition in bacteriology. International Journal of
among lipid-requiring diphtheroids from humans and characterization
Systemic Bacteriology, 44,
846 – 849.
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