1. business and industrial
2. agriculture and forestry
3. farms and ranches

Campylobacter in Finnish
poultry production in the 2000’s.
Ann-Katrin Llarena, ELL, PhD Student
Department of Food Hygiene and Environmental Health
University of Helsinki
EHYT/ Ann-Katrin Llarena/ Campylobacters in Finnish poultry
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Overview
•  Background
•  Chicken production in Finland
•  Methods
•  Results
•  Conclusions
EHYT/ Ann-Katrin Llarena/ Campylobacters in Finnish poultry
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Why study broilers?
•  Poultry production and products
‒  main source of human infection
‒  share genotypes w human infections
•  Chicken symptomless carry 10^8 CFU/g
caecum/small intestine
•  ! Campylobacter pos carcass (Sahin et al. 2002)
•  At retail
Why ?
Slaughterprocess
Defeathering and evisceration
!Fecal spill
Moist environment
‒  Spray cooling
‒  But: Air chilling (Finland)
•  only chicken products contaminated w/
relevant numbers of Campylobacter(EFSA)
Optimal ecological niche on
the chicken carcass
‒  Feather follicles
‒  Skin folds
Source: Ellerbroek et al., 2010,
Hermans 2012
EHYT/ Ann-Katrin Llarena/ Campylobacters in Finnish poultry
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Broiler production in Finland
Grandparents
•  Ross 308
•  Imported – quarantine
•  Approx 10 farms
Parent generation – the Breeders
•  Making of the broiler-egg
•  Approx 35 farms
Hatcheries
•  21 days
•  Approx 4 hatcheries
Rearing
•  Growing the broiler we buy
•  Approx 140 farms each year
•  Target point of this study
Source: Suomen Siipikarjanliitto ry & Suomen Broileryhdistys ry
EHYT/ Ann-Katrin Llarena/ Campylobacters in Finnish poultry
Slaughter
-  Atria (2 abattoirs)
-  Hk Scan (1 abattoir)
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Key numbers from
Rearing
Production
104,5mill kg 2014
Average
Avv
sized farm ≈40 000 birds
R
Raised in halls ≈15–20 000
35
5
35-39
days of growing
Alll-iin, All-out
A
All-outt : No
No depopulation
depopullattion
All-in,
28,8mill kg 1990
18.5kg
Consumption
7.6kg
Source: OSF:Luke, Meat production , www.mmm.fi, Gallup
Elintarviketieto,WP5.1 CamCon
EHYT/ Ann-Katrin Llarena/ Campylobacters in Finnish poultry
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Finnish monitoring
program of 2004
Maa- ja metsätalousministeriön asetus broilereiden
kampylobakteerivalvonnasta, Asetus nro 10/EEO/2007
•  Compulsory for all broiler slaughterhouses
•  Sample size:
•  All slaughter batches between June and October
•  Random testing rest of the year
•  10 intact ceca per slaughter batch
•  Pooled
•  Test for C. jejuni, C. coli and C. lari
Broiler batch: a group (or batch) of broilers of same age reared together in one
hall and slaughtered on the same day.
Source: Maa- ja metsätalousministeriön asetus broilereiden
kampylobakteerivalvonnasta,
Asetus nro 10/EEO/2007
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Aims
Baseline study
C. jejuni population in
Finnish broilers
Effects of year, season and
abattoir on C. jejuni
genotypes
EläinlEHYT/ Ann-Katrin Llarena/ Campylobacters in Finnish poultry
Link genotype and farm
information
evidence of an existing
persistent contamination
source?
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Methods
•  Cooperation with Evira – PhD Marjaana Hakkinen
‒  Only C. jejuni handled
‒  Isolates through the Finnish monitoring programme
•  MLST and PFGE (Evira) type strains from 5 years – 9 year period
‒  2004, 2006*, 2007*, 2008, 2012 (*de Haan et al. 2010)
•  Regression analysis:
‒  Outcome: ST or CC,
‒  Variables; Year, Month, Slaughterhouse
‒  (Cooperation w Anses, Ploufragan-Plouzané laboratory)
7894 batches
420 C. jejuni+
(3.1% (95% CI [2.1%, 4.1%]) )
380 M
MLST (90.5%)
•  Korzcak et al.
•  Miller et al.
•  WGS - Illumina
366 PFGE (87.1%)
•  SmaI
• w.
KpnI
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ww
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Results 1/4
Occurrence of C. jejuni
!
2004!
2006!
2007!
2008!
2012!
!"#$%
!"&$%
#"#$%
#"'$%
!"($%
&"&$)%
&"&$%
&"&$%
("*$%
+"#$%
Prevalence
June- October!
Prevalence November –
May subsequent year!
Seasonal distribution from all years
82-95% during
summer
Source: EFSA- Reports
EHYT/ Ann-Katrin Llarena/ Campylobacters in Finnish poultry
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Results 2/4
MLST – diverse population with
ST-45 CC dominant in all years
UA
21
45
<5%
S4, S55 ,
S66, S7
<5%
677
283
230
45
S64, S78
12 CCs
677
451
267
63 STs
03.04.2014
v
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Results 3/4
Regression analysis
45
677 21
Variable Year
ST-45 CC: More in 2004 and 2012
ST-21 CC: More in 2008
ST-677 CC: No effect
Outcome
ST-45 CC
ST-21 CC
Variable Season :
CC: No effect
ST-45: More from Sep
compared to spring
ST-677 CC
ST-45
Variable abattoir
ST-45 CC Less in B
ST-21 CC Less in A
ST-677 CC No effect
nim
n
imi /
im
Eläinlääketieteellinen tiedekunta / Henkilön nimi
Esityksen nimi
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Results 4/4
Farm data – GOOD SITUATION!
Approx 140 farms each year
Multiple batch farms
48.2% of these farms - within one rearing cycle
72.2% annualy did not
Often same genotype (MLST and PFGE)
deliver any C. jejuni pos.
House! house contamination
batches!
Multiple years:
60% of pos. only in one year Different genotypes each year
Most delivered just one pos
batch
Kuvaa ei voi näyttää. Tietokoneen muisti ei ehkä riitä kuvan
avaamiseen, tai kuva on vioittunut. Käynnistä tietokone uudelleen ja
avaa sitten tiedosto uudelleen.. Jos punainen
punainen x-merkki
x-merkki tulee edelleen
edelle
edell
näkyviin,
näkyvi
in, kuva
kuva on ehkä poistettava
poistet
poistet
oistettava
tava ja
ja lisättävä
lisättävä uudelleen.
uudelleen.
uudel
Kuvaa ei voi näyttää. Tietokoneen muisti ei ehkä riitä kuvan
avaamiseen, tai kuva on vioittunut. Käynnistä tietokone uudelleen ja
avaa sitten tiedosto uudelleen. Jos punainen x-merkki tulee edelleen
näkyviin, kuva on ehkä poistettava
tava ja
ja lisättävä
lisättävä uudelleen.
uudelleen.
ST-45 2004
NO!
ST-45 2006??
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Time-period!
Year!
2004!
Month!
June!
Farms !
A12!
A19!
%
B1!
ST-50/ S90S
ST-45*/ S66S%
July!
%
ST-45*/ S7%
Aug!
%
ST-677/ S8%
Sep!
%
ST-1539/ S96S
ST-3477/ K60S%
C5!
%
C8!
%
C28!
%
ST-451/ S5%
ST-230/ K36%
ST-230/ nd%
%
%
ST-230/ K59S
ST-677/ S64S%
%
ST-45/ S54S
ST-230/ K36S%
%
Oct!
%
%
%
ST-53/ S1S
ST-4596/ K78S%
%
%
%
Nov!
%
%
%
ST-4596/ K78 %
%
%
%
June!
ST-45/ S64%
%
ST-267/ S54S
ST-583/ S64S%
%
%
%
%
%
%
ST-3453/ S162S
ST-50/ S101S%
%
%
%
ST-586/ S13%
%
%
%
%
%
%
!
2006!
B5!
July!
Aug!
%
%
ST-267/ S64S
ST-267/ S102S
ST-267/ K36S
ST-45*/ S14S%
%
ST-267/ K36%
!
Sep!
ST-45/ S74S
ST-45/ S54S
ST-45/ S4S%
Oct!
%
%
RED: Carry over / same time rearing house!house / shared source
Yellow: Variance btw houses on a farm
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Conclusion
Take home messages
C. jejuni in chickens
•  Low prevalence of C. jejuni
on Finnish broiler farms
•  Month, year and abattoir
mildly influences the
C.jejuni population in
broilers
•  Diverse population but
predominant genotypes
•  ST-45 CC much more
common in Finland than
abroad
•  ST-677 CC Nordic
specialist?
On Farms
•  Effective hygiene barriers on
Finnish farms
•  ! rare and sporadic contamination
pattern of chicken flocks
•  No persistent contamination on
farms
•  From the environment
•  House to house do occur –
•  Only improvement point
Eläinlääketieteellinen tiedekunta / Henkilön nimi /
Esityksen nimi
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Aknowledgement
•  ELL Kaija Sivonen
RESEARCH ARTICLE
Predominant Campylobacter jejuni Sequence
Types Persist in Finnish Chicken Production
Ann-Katrin Llarena1*, Adeline Huneau2, Marjaana Hakkinen3‡, Marja-Liisa Hänninen1‡
1 Department of Food Hygiene and Environmental Health, Faculty of Veterinary Medicine, University of
Helsinki, Helsinki, Finland, 2 Anses, Ploufragan-Plouzané laboratory, BP 53, 22440, Ploufragan, France,
3 Food and Feed Microbiology Research Unit, Research and Laboratory Department, Finnish Food Safety
Authority, Evira, Helsinki, Finland
•  Ass. Professor Mirko
Rossi
‡ These authors contributed equally to this work.
* [email protected]
•  The research group
Abstract
OPEN ACCESS
Citation: Llarena A-K, Huneau A, Hakkinen M,
Hänninen M-L (2015) Predominant Campylobacter
jejuni Sequence Types Persist in Finnish Chicken
Production. PLoS ONE 10(2): e0116585.
doi:10.1371/journal.pone.0116585
Academic Editor: Igor Mokrousov, St. Petersburg
Pasteur Institute, RUSSIAN FEDERATION
Received: September 17, 2014
Accepted: December 11, 2014
Published: February 20, 2015
Copyright: © 2015 Llarena et al. This is an open
access article distributed under the terms of the
Creative Commons Attribution License, which permits
unrestricted use, distribution, and reproduction in any
medium, provided the original author and source are
credited.
Consumption and handling of chicken meat are well-known risk factors for acquiring campylobacteriosis. This study aimed to describe the Campylobacter jejuni population in Finnish chickens and to investigate the distribution of C. jejuni genotypes on Finnish chicken farms over a
period of several years. We included 89.8% of the total C. jejuni population recovered in Finnish poultry during 2004, 2006, 2007, 2008, and 2012 and used multilocus sequence typing
(MLST) and pulsed-field gel electrophoresis (PFGE) to characterize the 380 isolates. The typing data was combined with isolate information on collection-time and farm of origin. The C.
jejuni prevalence in chicken slaughter batches was low (mean 3.0%, CI95% [1.8%, 4.2%]), and
approximately a quarter of Finnish chicken farms delivered at least one positive chicken batch
yearly. In general, the C. jejuni population was diverse as represented by a total of 63 sequence types (ST), but certain predominant MLST lineages were identified. ST-45 clonal complex (CC) accounted for 53% of the isolates while ST-21 CC and ST-677 CC covered 11%
and 9% of the isolates, respectively. Less than half of the Campylobacter positive farms
(40.3%) delivered C. jejuni-contaminated batches in multiple years, but the genotypes (ST and
PFGE types) generally varied from year to year. Therefore, no evidence for a persistent C.
jejuni source for the colonization of Finnish chickens emerged. Finnish chicken farms are infrequently contaminated with C. jejuni compared to other European Union (EU) countries, making
Finland a valuable model for further epidemiological studies of the C. jejuni in poultry flocks.
Data Availability Statement: All relevant data are
within the paper and its Supporting Information files.
Funding: This work was supported by Finnish
Ministry of Agriculture and Forestry grant [No.
MMM2054/312/11] and by Coordination of European
Research on Emerging and Major Infectious
Diseases of Livestock (EMIDA ERA-NET) (Dnro 776/
312/2012) through the project entitled Biology and
Control of Campylobacter in the Chicken Supply
Chain (CamChain). The funders had no role in study
design, data collection and analysis, decision to
publish, or preparation of the manuscript.
Introduction
Campylobacter spp. is the most common causative agent for bacterial gastroenteritis worldwide, including Finland [1,2]. In 1999, campylobacteriosis surpassed the number of salmonellosis cases, and has since been the nation’s most important bacterial zoonosis with 4064
registered Finnish cases in 2013 [3,4]. The majority of cases (95%) are caused by Campylobacter
jejuni, which upon ingestion normally results in a self-limiting gastroenteritis, and the number
of cases peaks during the summer months of July and August [5].
PLOS ONE | DOI:10.1371/journal.pone.0116585 February 20, 2015
Eläinlääketieteellinen tiedekunta / Henkilön nimi /
Esityksen nimi
•  Funding:
•  Era-Net CamChain grant
•  Camcon
•  Finnish Ministry of
Agriculture and Forestry
1 / 18
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