Campylobacter antimicrobial resistance data · 2015. 1. 13. · Funded by the 7th Framework Programme of the European Union FP7 – Knowledge-based Bio-Economy (KBBE) Campylobacter

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



Union


(KBBE)

PROMISE: Protection of consumers by microbial risk mitigation

through combating segregation of expertise



Union


(KBBE)

WP

3 E

U f

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da

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ty

of p

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estim

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WP

1 N

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us ro

ute

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issio

n

WP

2 N

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WP

4 M

itig

atio

n o

f risk

thro

ugh

tra

inin

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Consumer protection

WP7 Management

WP

5 In

tera

ction

with

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WP

6 D

isse

min

ation

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Source: Promise project proposal, 2010



Union


(KBBE)

WP

3 E

U f

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odels

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nd

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WP 1 Neglected

exogenous routes

of pathogen

transmission

WP

Negle

cte

d

ind

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nou

s ro

ute

s o

f

tran

sm

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WP

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itig

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inin

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Consumer protection

WP7 Management

WP

5 In

tera

ction

with

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6 D

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min

ation

an

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Source: Promise project proposal, 2010



Union


(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



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(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



Testing

in lab 4

Testing

in lab 5 Testing

in lab 3

Testing

in lab 2




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(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




MLST typing



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(KBBE)

…seit Februar 2013

August 2012

December 2012

April 2013

February-May 2013


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



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(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)


of foodborne pathogens : Example – Vienna airport




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(KBBE)

Origin of passengers –

and food samples

Types of food items

Source: B. Appel, IAFP Promise meeting, 2014



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(KBBE)

Mainly dried or fermented meat or milk products...



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(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



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(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



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(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



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(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?



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(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



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(KBBE)



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(KBBE)



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(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



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(KBBE)



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(KBBE)



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(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



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(KBBE)



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(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



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(KBBE)



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(KBBE)

Another methodological approach…

..looking for something known



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(KBBE)

hens broilers

CZ CRO SI CRO SI HU



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(KBBE)

Metagenomic approach for searching of resistance genes…



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(KBBE)

Other issues: link antibiotic - biocide resistance?



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(KBBE)

MEHANIZEM MODULATORNEGA DELOVANJA – na nivoju GENOTIPA

Other issues: Mechanisms and modulation of antibiotic resistance?



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(KBBE)

Other issues: Adherence/biofilm formation – higher resistance



Union


(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



Union


(KBBE)

Acknowledgement

To AGES – C. Unger for invitation! To all of you for attention!

Documents

Campylobacter antimicrobial resistance data · 2015. 1. 13. · Funded by the 7th Framework Programme of the European Union FP7 – Knowledge-based Bio-Economy (KBBE) Campylobacter