campylobacter antimicrobial resistance data · 2015. 1. 13. · funded by the 7th framework...
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Funded by the 7th Framework
Programme of the European
Union
FP7 – Knowledge-based Bio-Economy
(KBBE)
Campylobacter antimicrobial resistance data
“Promise” project
Sonja Smole Možina
Campylobacter entlang der Lebensmittelkette
Vienna, October, 9th, 2014
Funded by the 7th Framework
Programme of the European
Union
FP7 – Knowledge-based Bio-Economy
(KBBE)
PROMISE: Protection of consumers by microbial risk mitigation
through combating segregation of expertise
Funded by the 7th Framework
Programme of the European
Union
FP7 – Knowledge-based Bio-Economy
(KBBE)
WP
3 E
U f
oo
d s
afe
ty
da
ta-m
odels
fo
r
un
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rtain
ty a
nd
va
ria
bili
ty
of p
reva
len
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estim
ate
s
WP
1 N
egle
cte
d
exo
ge
no
us ro
ute
s o
f
tra
nsm
issio
n
WP
2 N
egle
cte
d
ind
oge
nou
s ro
ute
s o
f
tra
nsm
issio
n
WP
4 M
itig
atio
n o
f risk
thro
ugh
tra
inin
g
Consumer protection
WP7 Management
WP
5 In
tera
ction
with
po
licy m
ake
rs
WP
6 D
isse
min
ation
an
d s
usta
ina
bili
ty
PROMISE: Protection of consumers by microbial risk mitigation
through combating segregation of expertise
Source: Promise project proposal, 2010
Funded by the 7th Framework
Programme of the European
Union
FP7 – Knowledge-based Bio-Economy
(KBBE)
WP
3 E
U f
oo
d s
afe
ty
da
ta-m
odels
fo
r
un
ce
rtain
ty a
nd
va
ria
bili
ty
of p
reva
len
ce
estim
ate
s
WP 1 Neglected
exogenous routes
of pathogen
transmission
WP
Negle
cte
d
ind
oge
nou
s ro
ute
s o
f
tran
sm
issio
n
WP
4 M
itig
atio
n o
f risk
thro
ugh
tra
inin
g
Consumer protection
WP7 Management
WP
5 In
tera
ction
with
po
licy m
ake
rs
WP
6 D
isse
min
ation
an
d s
usta
ina
bili
ty
PROMISE: Protection of consumers by microbial risk mitigation
through combating segregation of expertise
Source: Promise project proposal, 2010
Funded by the 7th Framework
Programme of the European
Union
FP7 – Knowledge-based Bio-Economy
(KBBE)
WP 1 Neglected exogenous routes of transmission
of foodborne pathogens
Studied contamination rate of food items carried by global
travellers (airports Vienna; Frankfurt; Bilbao)
Studied contamination rate of food items traded by small black
market in Romania
Studied contamination rate of food items traded at market selling
mainly homegrown food in Turkey
Studied contamination rate of food items carried away across
ground borders and seaports located in the Balkan countries
Studied contamination rate of food items imported to Greece
Source: M. Wagner, IAFP Promise meeting, 2014
Funded by the 7th Framework
Programme of the European
Union
FP7 – Knowledge-based Bio-Economy
(KBBE)
Number of foods items to be tested agreed
Harmonization of methods and sampling
Testing
in lab 1 Testing in
lab 6..
Admission
procedures
Collection of isolates
Characterisation
WP 1 Neglected exogenous routes of transmission
of foodborne pathogens
Testing
in lab 4
Testing
in lab 5 Testing
in lab 3
Testing
in lab 2
Source: M. Wagner, IAFP Promise meeting, 2014
Funded by the 7th Framework
Programme of the European
Union
FP7 – Knowledge-based Bio-Economy
(KBBE)
2500 foods items to be tested agreed
Salmonella
spp.
Listeria
monocytogenes
MLST typing
Virulotyping
Escherichia coli Campylobacter
jejuni/ coli
Staphylococcus
aureus
Micro array
MDR Verotoxin MDR
MDR
WP 1 Neglected exogenous routes of transmission
of foodborne pathogens
Source: M. Wagner, IAFP Promise meeting, 2014
MLST typing
Funded by the 7th Framework
Programme of the European
Union
FP7 – Knowledge-based Bio-Economy
(KBBE)
…seit Februar 2013
August 2012
December 2012
April 2013
February-May 2013
WP 1 Neglected exogenous routes of transmission
of foodborne pathogens : Example – German airports
Frankfurt Airport approx. 20 M passengers/ year (from third party countries)
2009 - 2011 ~0.3 % passengers (~ 50000/ year) inspected
approx. 7 % food items confiscated
approx. 2 kg food items per passenger
~ 2800 tons of food/ year Source: B. Appel, IAFP Promise meeting, 2014
Funded by the 7th Framework
Programme of the European
Union
FP7 – Knowledge-based Bio-Economy
(KBBE)
Côte d'Ivoire
IranNigeria
North Korea
Philippines
Russia
South Korea
Theiland
Ukraine
United Arab Emirates
Israel
Albannia
Tunisia
Serbia
Latvian
Qatar
Egypt 11%
China 11%
Vietnam
IndiaEthiopia
Jordan
Kazakhstan
Korea
Azerbaijan
Mongolia
MozambiqueBosnia and
Herzegovina
Armenia
South Africa
Montenegro
Kosovo
• Vienna International Airport
• August 2012 – March 2013
• 1473 products of animal origin found (6229 kg)
• 600 samples tested for pathogens and hygiene indicators
• 240 flights from 33 countries covered (61355 travellers)
WP 1 Neglected exogenous routes of transmission
of foodborne pathogens : Example – Vienna airport
Source: M. Wagner, IAFP Promise meeting, 2014
Funded by the 7th Framework
Programme of the European
Union
FP7 – Knowledge-based Bio-Economy
(KBBE)
Origin of passengers –
and food samples
Types of food items
Source: B. Appel, IAFP Promise meeting, 2014
Funded by the 7th Framework
Programme of the European
Union
FP7 – Knowledge-based Bio-Economy
(KBBE)
Mainly dried or fermented meat or milk products...
Funded by the 7th Framework
Programme of the European
Union
FP7 – Knowledge-based Bio-Economy
(KBBE)
Poultry less frequently found, if yes, then mostly processed...
Campylobacter very
rarely identified…
We got more than 120
presumptive samples,
but just a few strains
confirmed as C. jejuni
Funded by the 7th Framework
Programme of the European
Union
FP7 – Knowledge-based Bio-Economy
(KBBE)
Campylobacter
ENRICHMENT
PLATING-OUT
CONFIRMATION
Test portion (× g or × ml) in 9 × ml Preston or Bolton broth
mCCDA agar microaerophilic and aerobic atmosphere 44 4 h (41,5 °C 1 °C)
Detection of hippurate and indoxyl acetate
hydrolysis
Microaerophilic atmosphere 4-6 h (37 °C) and then
44 4 h (41,5 °C 1 °C)
Susceptibility on nalidixic acid and
cephalotin
Molecular (PCR, microarray),
immunological or latex agglut.test
mCCDA agar microaerophilic atmosphere
44 4 h (41,5 °C 1 °C)
Another isolation medium
Funded by the 7th Framework
Programme of the European
Union
FP7 – Knowledge-based Bio-Economy
(KBBE)
C. jejuni isolates from PROMISE were all isolated
from samples of fresh poultry from Balkan countries
ANTIBIOTIC RESISTANCE
MDR C. jejuni strains:
C2: resistant against STR, CIP, TET, ERY, NAL; Thr86Ala mutation,
tetO present
C33: resistant against STR, CIP, TET, NAL; Thr86Ala mutation;
without ERY R conferring mutation in 23S rRNA, tetO present
GENOTYPING METHODS
MLST (UL), flaA (UL), Binary multiplex PCR (mPCR) typing (UL), PFGE (VUW)
– C2: ST 1707, CC ST-607, flaA type14, mPCR clade C9ii
uncommon MLST sequence type and mPCR profile typical for wildlife environment
– C33: ST 400, CC ST-353, flaA 67, mPCR clade 5
Common MLST clonal complex and multiplex PCR profile, typical for farm animals
Funded by the 7th Framework
Programme of the European
Union
FP7 – Knowledge-based Bio-Economy
(KBBE)
0
50
100
150
200
250
2001 2002 2003 2004 2005 2006 2007 2008 2009 2010 2011 2012
Št.
po
trje
nih
prim
ero
v x
10
00
Human campylobacterioses 2001-12 in EU
Why to talk more about C. jejuni resistance in EU?
Funded by the 7th Framework
Programme of the European
Union
FP7 – Knowledge-based Bio-Economy
(KBBE)
0
10
20
30
40
50
60
70
80
Campylobacter spp. C. jejuni C. coli
Št.
od
po
rn
ih s
ev
ov
v %
CIP
ERY
GEN
NAL
TET
Average for EU, 2012
Funded by the 7th Framework
Programme of the European
Union
FP7 – Knowledge-based Bio-Economy
(KBBE)
Funded by the 7th Framework
Programme of the European
Union
FP7 – Knowledge-based Bio-Economy
(KBBE)
Funded by the 7th Framework
Programme of the European
Union
FP7 – Knowledge-based Bio-Economy
(KBBE)
Funded by the 7th Framework
Programme of the European
Union
FP7 – Knowledge-based Bio-Economy
(KBBE)
0
10
20
30
40
50
60
70
80
90
C. jejuni C. coli
Št.
od
po
rn
ih s
ev
ov
v %
CIP
ERY
GEN
NAL
TET
Average for EU, 2012
0
10
20
30
40
50
60
70
2009 2010 2011 2012
Št.
od
po
rn
ih s
ev
ov
v %
CIP
ERY
GEN
NAL
TET
Average for EU 2009-12
Funded by the 7th Framework
Programme of the European
Union
FP7 – Knowledge-based Bio-Economy
(KBBE)
Funded by the 7th Framework
Programme of the European
Union
FP7 – Knowledge-based Bio-Economy
(KBBE)
Funded by the 7th Framework
Programme of the European
Union
FP7 – Knowledge-based Bio-Economy
(KBBE)
Funded by the 7th Framework
Programme of the European
Union
FP7 – Knowledge-based Bio-Economy
(KBBE)
111 samples of retail fresh chicken meat; ISO 10272 kvali/kvanti: 56% Campy pos., ~10-100 CFU/g,
9 PFGE types, 11 MLST types (most frequent CC-21, CC-353, CC-354).
Antibiotc resistance profile of C. jejuni isolates (N=35) Source: CRP V4-1110)
GEN CIP TET ERY NAL CHL STR
R 0 27 2 1 21 0 1
S 35 8 33 34 14 35 34
R (%) 0 77 5.7 2.8 60 0 2.8
S (%) 100 23 94.3 97.2 40 100 97.2
2012-2013 (sampling Nov 12- Jun 13)
22,20%
70,40%
3,70%3,70%
0R
1R
2R
3R
Slovenia
Funded by the 7th Framework
Programme of the European
Union
FP7 – Knowledge-based Bio-Economy
(KBBE)
Funded by the 7th Framework
Programme of the European
Union
FP7 – Knowledge-based Bio-Economy
(KBBE)
Antibiotic resistance gene spreading
Expansion of resistant clones
Clones resistant to ciprofloxacin have a
resistance conferring point mutation in
QRDR region of gyrA gene
Determining genetic relatednes based
on QRDR sequence and MLST type
Molecular epidemiology of Campylobacter
Funded by the 7th Framework
Programme of the European
Union
FP7 – Knowledge-based Bio-Economy
(KBBE)
Funded by the 7th Framework
Programme of the European
Union
FP7 – Knowledge-based Bio-Economy
(KBBE)
Another methodological approach…
..looking for something known
Funded by the 7th Framework
Programme of the European
Union
FP7 – Knowledge-based Bio-Economy
(KBBE)
hens broilers
CZ CRO SI CRO SI HU
Funded by the 7th Framework
Programme of the European
Union
FP7 – Knowledge-based Bio-Economy
(KBBE)
Metagenomic approach for searching of resistance genes…
Funded by the 7th Framework
Programme of the European
Union
FP7 – Knowledge-based Bio-Economy
(KBBE)
Other issues: link antibiotic - biocide resistance?
Funded by the 7th Framework
Programme of the European
Union
FP7 – Knowledge-based Bio-Economy
(KBBE)
MEHANIZEM MODULATORNEGA DELOVANJA – na nivoju GENOTIPA
Other issues: Mechanisms and modulation of antibiotic resistance?
Funded by the 7th Framework
Programme of the European
Union
FP7 – Knowledge-based Bio-Economy
(KBBE)
Other issues: Adherence/biofilm formation – higher resistance
Funded by the 7th Framework
Programme of the European
Union
FP7 – Knowledge-based Bio-Economy
(KBBE)
• Risk for antibiotic resistance development and spreading is high!
• Antibiotic resistance of C. jejuni/coli is frequent, especially against CIP in fresh
chicken meat – still increasing in central and south EU countries.
• Responsible use of antimicrobial agents is urgently needed!
• Support to research and development of new antimicrobials and alternative agents
and strategies should be given!
• Support to general education of consumers, producers and other food-chain
stakeholders is needed!
Conclusions –urgent actions needed
Funded by the 7th Framework
Programme of the European
Union
FP7 – Knowledge-based Bio-Economy
(KBBE)
Acknowledgement
To AGES – C. Unger for invitation! To all of you for attention!