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4 AUTHORS, INCLUDING:
Sonja Smole Moina
Anja Klancnik
University of Ljubljana
University of Ljubljana
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Review
Campylobacter and
its multi-resistance in
the food chain
Sonja Smole Mo
zina*,
Marija Kurin
ci
c, Anja Klan
cnik
and Ana Mavri
University of Ljubljana, Biotechnical Faculty,
Department of Food Science and Technology,
Jamnikarjeva 101, SI-1000 Ljubljana, Slovenia
(e-mail: sonja.smole@bf.uni-lj.si)
Increasing antimicrobial resistance is an urgent world-wide
problem, including multidrug resistant microorganisms transmitted via the food chain. Campylobacteriosis is the leading
bacterial food-borne illness and most frequently reported zoonosis in humans. Despite the differences in results and some
insufficiency in the methodological harmonization among reports from different countries, there was an evident increase in
prevalence and multiple resistance of Campylobacter in food
production environments in central and southern EU-member
states (MSs). Non-specific efflux pumps are involved in bacterial reduced susceptibility and/or resistance against antibiotics
and other unrelated antimicrobials, including bile salts. Beside
this, the synergistic antimicrobial activity of some compounds
which could act as inhibitors of efflux pumps is a challenge to
develop more efficient protection against food-borne pathogens. Testing the activity of known and putative natural efflux
pump inhibitors (EPIs) in Campylobacter resistant isolates revealed new information about the mechanisms involved in resistance and new approaches for Campylobacter risk
management.
Introduction
Antimicrobials are extremely important for treatment
and disease prevention in animal livestock production.
For almost 50 years and until their recent withdrawal in
the EU, antibiotics at subtherapeutic levels were also used
* Corresponding author.
0924-2244/$ - see front matter 2010 Elsevier Ltd. All rights reserved.
doi:10.1016/j.tifs.2010.09.003
for growth promotion, improved feed efficiency and enhanced performance of livestock (Donnelly, 1999). However, it is generally acknowledged that the use of
antimicrobials in food animals drives the emergence of antimicrobial resistance and promotes its dissemination via
the food chain. The development and spread of antimicrobial resistance of zoonotic bacteria with its reservoirs in
healthy food animals, e.g. poultry, pigs and cattle, has become a global public health problem (EFSA, 2008).
Antimicrobial resistance in the food chain, including
multidrug resistance, is a complex issue. To understand
the public health concern about antimicrobial resistance
in zoonotic bacteria, not only the use of antimicrobials
but other factors, including bacterial virulence and capacity
to spread, the genetic basis of antimicrobial resistance, and
co- and cross-resistance, as well as the rate of bacterial
transfer, dependent on the animal and bacterial species,
slaughter practices and production forms should also be
considered. In a very recent joint opinion of EFSA, the
European Centre for Disease Control and Prevention
(ECDC), the European Medicines Agency (EMEA) and the
European Commissions Scientific Committee on Emerging
and Newly Identified Health Risks (SCENIHR) on antimicrobial resistance focused on zoonotic infections (EFSA,
2009a), some combinations of bacterial agent/antimicrobial/animal species are considered of the highest concern.
Among microbes these are Campylobacter and Salmonella,
and among antimicrobials quinolones, cephalosporines and
macrolides. Cephalosporins are not used for campylobacterioses treatment, so only quinolone, macrolide and multi-resistance of Campylobacter are targeted in this review. In
selecting the segment of the food chain with the highest emphasis on human health, chicken meat production was highlighted for several reasons: i) the higher prevalence of
Campylobacter in chicken than in other meats, ii) usage of
antibiotics on livestock farms is increasing in several EUMSs, especially in broiler production (MARAN, 2007), iii)
the high and still increasing consumption of chicken meat because of consumer preferences for white meat and the competitive prices of poultry with respect to other meats
(European Commission Prospects, 2009).
The aim of this review is to illustrate recent trends in the
prevalence and antibiotic (multi)resistance of Campylobacter jejuni and Campylobacter coli in the food chain. Beside
this, the role of efflux in multiple resistance of Campylobacter is highlighted, not only to antibiotics but also for
60
50
2003
40
2004
30
2005
20
2006
10
2007
Farm animals
Beef
Pork
Chicken
Cattle
Pigs
Broilers
Retail meat
100
90
80
70
60
50
40
30
20
10
0
Broiler flocks
Chicken meat
Es
to
n
N ia
or
wa
Sw y
e
D den
en
m
ar
La k
tv
ia
Sp
ai
I n
Sw rela
itz nd
er
la
n
Au d
st
C Ge ria
ze
ch rma
Re ny
pu
Sl blic
ov
en
ia
Ita
Fr ly
an
S l ce
ov
ak
ia
S.S. Mo
z ina et al. / Trends in Food Science & Technology 22 (2011) 91e98
92
5%
6%
12%
21%
18%
21%
93
21%
0R
16%
1R
2R
36%
3R
23%
38%
Human isolates ,
N = 636
32%
Animal isolates,
N = 246
>3R
22%
31%
Fig. 3. Multiresistance of Campylobacter jejuni/coli isolates from human clinical samples, broiler flocks and retail chicken meat samples (source:
EFSA country reports for A, D, HU and SI, 2007 and 2009).
94
et al., 2010). Similar results were found when we tested retail meat and human clinical samples in two geographical
areas (Slovenia and Bosnia and Herzegovina) already in
the period 2001e2003 (Kurincic et al., 2009; Smole
Mozina, Zorman, & Berce, 2005).
In Fig. 3, combined data about the multiple antibiotic resistance of C. jejuni/coli isolates from chicken meat and
other sources (animal, human isolates) reported to EFSA
in the years 2006e2007 from four central European countries are presented (Data from Austria, Germany, Hungary
and Slovenia were included). For all of them resistance to
aminoglycosides (gentamicin), fluoroquinolones (enrofloxacin), macrolides (erythromycin), quinolones (nalidixic
acid), penicillins (ampicillin) and tetracyclines (oxytetracyclin) was tested. Much higher prevalence of multiresistant
strains is again evident for the food chain, especially
meat isolates, in comparison to human isolates.
Table 1. (Multi)resistance (minimal inhibitory concentration (MIC, mg/mL or mg/mL) in bold means high resistance) of ten Campylobacter isolates from the food chain and the efficiency (U) of efflux pump inhibitors (EPIs, PAbN (20 mg/mL) and NMP (80 mg/mL)) in reducing the resistance to erythromycin, ciprofloxacin, bile salts and Na-deoxycholate.
Strain
803
FC8
722
725
375
M37
171
7114
1076
140
ATCC 33559
ATCC 33560
Source
Species
Water jejuni
Poultry coli
Pig
coli
Pig
coli
Poultry jejuni
Poultry coli
Poultry coli
Pig
coli
Pig
coli
Poultry coli
Feces, coli
pig
Feces, jejuni
bovine
Erythromycin
Ciprofloxacin
Bile salts
Na-deoxycholate
MIC
mg/mL
U-PAN
U-NMP
MIC
mg/mL
U-PAN
U-NMP
MIC
mg/mL
U-PAN
U-NMP
MIC
mg/mL
U-PAN
U-NMP
1
4
8
8
64
512
512
>512
>512
>512
2
16
32
64
128
32
32
16
32
32
32
32
32
8
1
1
16
4
4
512
32
8
32
2
64
128
0.25
128
2
16
8
64
32
0.063
1
2
64
1
64
1
1
1
64
1
1
1
4
64
2
64
2
2
2
64
4
1
64
64
64
16
16
16
16
16
64
64
64
2
4
2
4
2
1
4
4
64
1
32
4
64
16
8
4
2
8
16
64
2
32
64
64
16
64
8
32
16
2
32
16
32
4
1
1
8
8
1
16
4
64
1
4
32
64
2
32
32
32
16
4
32
2
64
0.25
64
32
64
32
A four-fold or greater reduction of MIC after EPI addition is considered significant and indicated in bold.
MIC EGCG
8
16
4
8
4
64
16
64
16
32
16
1
8
1
1
8
2
32
1
1
1
128
64
8
0.25
1
1
1
1
8
2
1
1
1
4
0.5
64
4
>512
0.25
jejuni
jejuni
coli
coli
jejuni
Poultry
Poultry
Pig
Pig
Human
K49/4
375
FC8
7114
NCTC
11168
U-*0.5
8
32
16
8
32
64
8
64
2
64
4
16
16
2
16
U-0.25
U-0.5
U-0.5
32
32
16
8
32
MIC
U-0.25
EGCG
MIC
U-0.5
EGCG
MIC
mg/mL
EGCG
EGCG
EGCG
MIC
mg/mL
U-*
0.5 MIC
U-**
0.25 MIC
MIC
mg/mL
EGCG
MIC
U-0.25
MIC
MIC
mg/mL
EGCG
MIC
EGCG
MIC
Na-deoxycholate
Bile salts
Ciprofloxacin
Erythromycin
Species
Source
Future challenges
We are currently studying several approaches to confirm
the specific role of Campylobacter efflux pumps via testing
well-known EPIs in isolates from the food chain and in the
mutants in specific efflux pump genes. Another important
subject is the involvement of efflux in resistance to other
95
Strain
Table 2. The efficiency (U) of the plant phenolic compound, (L)epigallocatechin gallate (EGCG) as a putative efflux pump inhibitor in Campylobacter isolates in reducing the resistance to
erythromycin, ciprofloxacin, bile salts and Na-deoxycholate. Four-fold or greater reduction of MIC is considered significant and indicated in bold (MICs of EGCG for the isolates K49/4 and
375 were 0.078, for FC8 was 0.625, but for the isolates 7114 and 11168 were 0.313 mg/mL).
S.S. Mo
z ina et al. / Trends in Food Science & Technology 22 (2011) 91e98
S.S. Mo
z ina et al. / Trends in Food Science & Technology 22 (2011) 91e98
antimicrobials frequently used in the food chain, e.g. disinfectants, cleaning and decontamination agents, to contribute to knowledge about a possible link between the
resistance to biocides and antibiotics, which has been recently confirmed in some other enteric pathogens
(Karatzas et al., 2007; Thorrold, Letsoalo, Duse, &
Marais, 2007). However, the most challenging task is
screening for natural antimicrobial and resistance modifying agents, preferably from plants because of their GRAS
status (Klancnik, Guzej, Kolar, Abramovic, & Smole
Mozina, 2009). In Table 2, the activity of the plant phenolic
compound ()-epigallocatechin gallate (EGCG) as a putative efflux pump inhibitor and resistance modifying agent
in selected Campylobacter isolates is presented. Some
strains (e.g. C. coli VC7114 and FC8 and C. jejuni 375)
are the same as in Table 1, so also comparison with the activity of known EPIs, PAbN and NMP is possible. To our
knowledge this is the first evidence of the activity of any
plant phenolic compound as resistance modifying agent in
Campylobacter for some non-related antimicrobials. This
could have very useful applications, but should be further
studied with higher number of natural isolates and their mutants in target efflux genes both in in vitro and in vivo
systems.
Acknowledgement
We are grateful to Jasna Kovac for help in preparing
some figures and The Slovenian Research Agency
(ARRS) for PhD grants for M.K. and A.M., a post-doc
grant to A.K. and the financing of our national and bilateral
projects. This work was also partially financed by FP6036272.
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