Impotentie brengt een constant ongemak met zich mee, net als fysieke en psychologische problemen in uw leven cialis kopen terwijl generieke medicijnen al bewezen en geperfectioneerd zijn
Journal of Food Protection, Vol. 71, No. 3, 2008, Pages 602–607
Copyright ᮊ, International Association for Food Protection
Comparison of Antimicrobial Resistance of
Campylobacter jejuni and Campylobacter coli Isolated from
Humans and Chicken Carcasses in Poland
˙ BIETA ROZ˙YNEK,
1* KATARZYNA DZIERZ˙ANOWSKA-FANGRAT,
1 DOROTA KORSAK,
2,3 PIOTR KONIECZNY,
5 JOLANTA SZYCH,
5 MIROSŁAW JAROSZ,
2 AND DANUTA DZIERZ
1Department of Clinical Microbiology & Immunology, The Children’s Memorial Health Institute, 04-730 Warsaw, Poland;
2National Food and Nutrition Institute, Warsaw, Poland;
3Department of General Microbiology, Faculty of Biology, University of Warsaw, Warsaw,
4Infectious Diseases Hospital, Dziekanow Lesny, Warsaw, Poland; and
5National Institute of Hygiene, Warsaw, Poland
MS 07-048: Received 29 January 2007/Accepted 11 September 2007
-associated gastroenteritis remains an important cause of morbidity worldwide, and some evidence suggests
that poultry is an important source of this foodborne infection in humans. This study was conducted to analyze the prevalenceand genetic background of resistance of 149 Campylobacter jejuni
and 54 Campylobacter coli
strains isolated from broilerchicken carcasses and from stool samples of infected children in Poland from 2003 through 2005. Nearly all isolates weresusceptible to macrolides and aminoglycosides. The highest resistance in both human and chicken strains was observed forciprofloxacin (more than 40%), followed by ampicillin (13 to 21%), and tetracycline (8 to 29%). Resistance to ampicillin andtetracycline rose significantly between 2003 and 2005. Slight differences in resistance between human and chicken isolatesindicate that although chicken meat is not the only source of Campylobacter
infection in our population, it can be involvedin the transmission of drug-resistant Campylobacter
strains to humans.
and Campylobacter coli
and avilamycin have been used as growth promoters in
portant causes of foodborne gastroenteritis in humans in
poultry farming operations, but this procedure was banned
many developed countries (2, 7, 24).
Surveillance data on
in 2006 according to the European Community 90/167/EEC
human campylobacteriosis in Poland are limited. However,
However, antimicrobials such as tylosin, tia-
the results of a 6-year study in Polish children with diarrhea
mulin, lincomycin, amoxicillin, ampicillin, and tetracy-
revealed that the incidence of Campylobacter
clines can be used in medicated feed (6, 13).
ceeded that of salmonellosis (32).
Campylobacteriosis is of-
The most common mechanism of high-level quinolone
ten associated with handling of raw poultry or eating of
resistance in Campylobacter
spp. is associated with a mu-
undercooked poultry meat (4).
Cross-contamination of raw
tation in the quinolone resistance-determining region at po-
poultry to other ready-to-eat foods via the cook’s hands or
sition 86 in the gyrA
The most frequently ob-
kitchen utensils also has been reported (27).
served molecular mechanism of tetracycline resistance is
teriosis is generally a self-limiting disease. However, in cas-
the production of Tet(O), a ribosomal protection protein.
es of severe or extraintestinal infections and in immuno-
(O) gene is often associated with conjugative plas-
compromised patients, antibiotics may be required. Eryth-
mids. However, this gene also can be chromosomally lo-
romycin is the usual drug of choice for the treatment of
The high-level resistance to erythromycin is me-
diated by mutations in domain V of the 23S rRNA gene at
gentamicin, and tetracycline also are clinically effective in
infections when antimicrobial ther-
The aim of this study was to compare the prevalence
and genetic background of antimicrobial resistance in Pol-
In many studies, a significant rise in resistance to fluo-
ish strains of C. jejuni
and C. coli
isolated from chicken
roquinolones, tetracycline, and erythromycin has been dem-
carcasses and children to determine whether there was a
onstrated in C. jejuni
and C. coli
isolates (1, 15, 21, 22,
31, 34, 35).
Increasing resistance is probably partly due to
MATERIALS AND METHODS
the wide use of these antimicrobials in veterinary medicine,especially in poultry (9, 10, 15, 40).
In Poland, flavomycin
Samples of 130 broiler chicken carcasses were obtained from
selected supermarkets in Warsaw. Stool samples from 946 chil-dren with diarrhea were obtained from three large pediatric hos-
* Author for correspondence. Tel: ϩ48 22 815 72 70; Fax: ϩ48 22 815
pitals in Warsaw. All samples were collected from 2003 through
72 75; E-mail: firstname.lastname@example.org.
ANTIBIOTIC RESISTANCE OF CAMPYLOBACTER
ISOLATES FROM POLAND
Isolation of Campylobacter
spp. from chicken meat was per-
resistance to ampicillin rose from 8 to 35.5% (P
formed according to the International Organization for Standard-
in human isolates and from 5.8 to 30.4% (P
ϭ 0.01) in
ization guideline 10272 (20).
After initial bacterial characteriza-
chicken isolates. High rates of ciprofloxacin resistance
tion by phase-contrast microscopy, Gram staining, catalase and
(Ͼ40%) were noted in both human and chicken isolates
oxidase production, and growth at 25 and 42ЊC, suspected colo-
and did not change significantly during the study period.
nies were confirmed as C. jejuni
or C. coli
using the PCR method
Cross-resistance to nalidixic acid and ciprofloxacin was
with specific primers (33).
Isolation and identification of human
found in all quinolone-resistant strains.
strains were performed according to World HealthOrganization recommendations (17)
and confirmed by the PCR
Resistance phenotypes are shown in Table 3. Resis-
isolates were maintained in 20%
tance to quinolones was the predominant phenotype ob-
glycerol in Brucella
broth at Ϫ70ЊC for subsequent analysis.
served both in human (50%) and chicken (63.3%) isolates.
and C. coli
susceptibility to seven antimicrobials
The frequency of double resistance was significantly higher
was determined by the Etest (AB Biodisk, Solna, Sweden) on
Ͻ 0.001) in human isolates (27 of 68 isolates, 39.7%)
Mueller-Hinton agar containing 5% sheep blood. Etests were used
than in chicken isolates (7 of 60 isolates, 11.6%). Only two
in accordance with the manufacturer’s instructions. The plates
human C. jejuni
isolates were simultaneously resistant to
were incubated at 37ЊC for 48 h under microaerophilic conditions.
The following CLSI interpretative criteria for the Enterobacteri-
All 149 C. jejuni
and 54 C. coli
strains were screened
family were used as breakpoints for Campylobacter
for molecular mechanisms of resistance to tetracycline,
tance: 16 mg/liter gentamicin and tetracycline, 32 mg/liter nali-
erythromycin, and ciprofloxacin. The results of phenotypic
dixic acid, 4 mg/liter ciprofloxacin, and 32 mg/liter ampicillin.
For erythromycin and azithromycin, an interpretative breakpoint
and genetic analyses of resistance to tetracycline were fully
spp. of 8 mg/liter was used (26). C. jejuni
concordant. All tetracycline-resistant isolates possessed the
ATCC 33560 and C. coli
ATCC 33559 were used as reference
(O) gene. The results of macrolide susceptibility testing
by Etests and 23S rRNA gene analysis were in agreement
(O) gene was detected by PCR assay (12).
for all but one isolate. The only C. jejuni
mutations in the gyrA
gene were identified by the mismatch am-
to erythromycin by the Etest method (MIC of 24 mg/liter)
plification mutation assay PCR (MAMA PCR) as described else-
had no detectable A2074C or A2075G mutations in the 23S
where (41, 42)
; however, different reaction conditions were used:
rRNA gene by either PCR-RFLP assay or direct sequenc-
94ЊC for 5 min for predenaturation followed by 35 cycles of 94ЊC
ing. All Campylobacter
isolates resistant to quinolones by
for 30 s, 54ЊC for 1 min, and 72ЊC for 1 min and a final extension
the Etest had Thr-86-Ile mutations in the gyrA
at 72ЊC for 7 min. The presence of Thr-86-Ile substitutions was
tected by MAMA PCR and PCR-RFLP assays. However,
additionally confirmed by the PCR–restriction fragment lengthpolymorphism (RFLP) method (3).
The PCR-RFLP method also
these mutations were also found by both molecular methods
was used for the detection of A2074C and A2075G mutations in
in two isolates phenotypically susceptible to ciprofloxacin
the 23S rRNA gene (39).
Isolates with discrepant phenotypic and
(both MICs of 2 mg/liter) and nalidixic acid (MICs of 1
genetic susceptibility results were subjected to direct sequencing
and 8 mg/liter). Both isolates were further confirmed by
of respective genes using DYEnamic ET Terminator Cycle Se-
direct sequencing to contain Thr-86-Ile mutations.
quencing Kit (GE Healthcare, Chalfont St. Giles, UK) on an Ap-plied Biosystems 3730 automated sequencer (Applied Biosystems,
In several studies, handling and consumption of poul-
The resistance rates were compared us-
try has been identified as important risk factors for Cam-
infections in humans (4, 16, 18, 36, 37).
Thefrequency of contamination of retail chicken meat with
spp. has exceeded 80% in some countries
During a 3-year study, 100 Campylobacter
(4, 36, 37).
In a recent survey undertaken in Poland by the
isolated from 130 chickens, and 103 strains were obtained
National Food and Nutrition Institute, 75% of chicken car-
from 946 children. C. jejuni
comprised 86.4% of human
casses were contaminated with Campylobacter
ϭ 89) and 60% of chicken isolates (n
published data). Microorganisms that colonize food animals
The remaining isolates were C. coli.
or contaminate food products and are transmitted to humans
The results of antimicrobial susceptibility testing are
via the food chain also can serve as a source of antimicro-
summarized in Tables 1 and 2. All chicken Campylobacter
bial resistance genes for bacteria residing in the human gas-
strains and all but one human strain were susceptible to
trointestinal tract. Increasing Campylobacter
macrolides. Resistance to gentamicin was detected in three
antimicrobials is likely the result of the wide use of these
strains isolated from children and in one chicken
agents in veterinary medicine and as growth promoters in
strain. All gentamicin-resistant strains were obtained in
animal farming operations. An estimated 50% of all anti-
2005. The overall resistance to tetracycline in chicken C.
biotic usage worldwide is in veterinary medicine (24).
and C. coli
strains was 8.3 and 10%, respectively.
of quinolones, mainly enrofloxacin in veterinary applica-
The respective rates in human isolates were 15.7 and
tions, has been associated with the emergence of ciproflox-
28.5%. Tetracycline resistance in chicken isolates increased
acin resistance in poultry and human strains of Campylo-
from 0% in 2003 to 17.3% in 2005 (P
ϭ 0.01). Ampicillin
bacter (9, 10, 15).
In contrast, in countries where quino-
and ciprofloxacin had low activity against C. jejuni
lones have not been used in food-producing animals, very
strains regardless of their origin. From 2003 to 2005,
low incidences (2%) of ciprofloxacin-resistant Campylo-
TABLE 1. MIC distributions of seven antimicrobial agents for 149
C. jejuni and 54
C. coli isolatesa
AM, ampicillin; Cj, C. jejuni
; H, human; Ch, chicken; Cc, C. coli
; EM, erythromycin; AZ, azithromycin; TC, tetracycline; CI,
ciprofloxacin; NA, nalidixic acid; GM, gentamicin.
have been observed (24).
In Poland, quinolones are
86-Ile substitutions also were found in isolates resistant to
allowed for therapeutic use in veterinary medicine by the
ciprofloxacin but fully susceptible to nalidixic acid (5).
Ministry of Agriculture and Rural Development, which
A high prevalence of macrolide resistance in Campylo-
may explain the high resistance rates in Campylobacter
has been reported mainly in those European coun-
strains analyzed in this study and in Salmonella
tries where tylosin has been used as an animal growth pro-
strains of poultry origin (19).
The high preva-
Among 203 Campylobacter
isolates examined in
lence of ciprofloxacin resistance in Campylobacter
this study, only 1 clinical C. jejuni
strain had low resistance
obtained from children may result from transmission of
to erythromycin and azithromycin by the Etest, and this
poultry strains, because quinolones are used only occasion-
strain was simultaneously resistant to tetracycline, cipro-
ally in children, mainly in the treatment of cystic fibrosis.
floxacin, and ampicillin. However, this strain did not con-
spp. resistance to quinolones most fre-
tain either of the most frequent mutations in the 23S rRNA
quently results from the Thr-86-Ile mutations in the gyrA
gene responsible for macrolide resistance, which suggests
gene (ACA→ATA in C. jejuni
and ACT→ATT in C. coli
that resistance was associated with the efflux pump. Such
In this study, all isolates resistant to quinolones
a mechanism has been described in multidrug-resistant
by the Etest possessed Thr-86-Ile substitutions. However,
isolates with low-level macrolide resistance
these mutations also were found in two isolates phenotyp-
ically categorized as quinolone susceptible (ciprofloxacin
An increase in the resistance of Campylobacter
MIC of 2 mg/liter for both isolates and nalidixic acid MICs
racyclines has been observed in many countries. The prev-
of 1 and 8 mg/liter). A similar phenomenon has already
alence of tetracycline resistance among Campylobacter
been observed. The gyrA
Thr-86-Ile mutations were re-
lates in the United States and Germany is close to 50% (23,
ported in Campylobacter
isolates with a ciprofloxacin MIC
and in Spain it exceeds 70% (34).
In this study, overall
of 2 mg/liter, which suggests that this concentration could
tetracycline resistance did not exceed 30%, and there were
be a more appropriate breakpoint for this agent (5, 29).
no significant differences in tetracycline resistance between
ANTIBIOTIC RESISTANCE OF CAMPYLOBACTER
ISOLATES FROM POLAND
human and chicken isolates. However, a significant increase
tern in epidemiological investigations (31, 34, 35, 40).
in resistance was found during the last 3 years in chicken
most frequent resistance pattern observed in this study was
strains (but not in human isolates). This increase may be
the lack of susceptibility to a single agent, which was char-
associated with the use of chlortetracycline and doxycycline
acteristic of 83.3 and 56% of chicken and human strains,
in medicated feed for poultry and swine (6).
The most im-
respectively. Double resistance was detected three times
portant mechanism of tetracycline resistance in Campylo-
more frequently in human strains than in chicken strains.
is the plasmid-mediated transfer of the tet
Slight differences in resistance between human and chicken
which codes for the ribosomal protection protein (12).
strains may indicate that chickens are not the only source
our tetracycline-resistant isolates possessed the tet
infection in our population. This finding
Some authors have proposed the use of resistance pat-
is concordant with our previous findings of a different dis-
TABLE 2. Resistance of
Campylobacter isolates of human and chicken origin to five antimicrobial agents in 2003, 2004, and 2005
In 2003, 2004, and 2005, 25, 47, and 31 human isolates and 51, 26, and 23 chicken isolates, respectively, were obtained.
TABLE 3. Resistance phenotypes of chicken and human
CIR, resistance to ciprofloxacin; TCR, resistance to tetracycline; AMR, resistance to ampicillin; GMR, resistance to gentamicin; EMR,
tribution of some virulence markers in human and chicken
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J Oral Pathol Med (2004) 33: 445–50ª Blackwell Munksgaard 2004 Æ All rights reservedOral histoplasmosis associated with HIV infection:a comparative studyS. L. Herna´ndez1, S. A. Lo´pez de Blanc1,2, R. H. Sambuelli3,4, H. Roland3, C. Cornelli3, V. Lattanzi1,M. A. Carnelli41Clinical Stomatology B, School of Dentistry, National University of Co´rdoba, Co´rdoba; 2Department of Oral Pathol
CLASSE DES PRODUITS DOPANTS 1. Les stimulants 2. Les analgésiques narcotiques 3. Les anabolisants 4. Les corticostéroïdes 5. Les hormones peptidiques et analogues 6. Les bêta-bloquants 7. Les diurétiques 1. Les stimulants De quoi s’agit-il? Les stimulants agissent surtout sur le système nerveux central et cardiovasculaire. Ils présentent la particularité psychol