Lecture 2 Campylobateria

Campylobacter
Alejandra Culebro
Department of Food Hygiene and Environmental Health

Faculty of Veterinary Medicine
www.helsinki.fi/yliopisto
Overview
Campylobacteriosis
The genus Campylobacter
History
Taxonomy
Detection and isolation methods

Typing methods
Emerging campylobacters
Confirmed cases per 100 000 population
90
80
70
60
50
40
30
Average of EU member states except for Latvia, Portugal,
Romania, and Slovenia
Finland
20
10
0
2007
2008
2009
Source: EFSA 2009; EFSA 2010; EFSA 2011; EFSA and ECDC 2012; EFSA and ECDC 2013
Salmonella
Source: National Institute for Health and Welfare 2011
2010
2011
Year
Campylobacter
30.10.2013
Campylobacter
Source: THL
Source: EFSA 2010; EFSA 2011

200
180
160
140
120
100
80
60
40
20
0
Fin
EU
US
CR
New
Zealand
UK
Symptoms
500-800 cells
3.2 days
Self-limiting
Relapse 5-10%
- Ingested dose
- Strains virulence
- Acquired immunity
- Health status
Yet
Source: Black and others 1988; Feodoroff and others 2011; Pacanowski 2008;
Skirrow 1977
Treatment and prevention
Treatment:
No treated
BUT:
Erythromycin (Acheson and Allos 2001;
Taylor and Tracz 2005)
Macrolides, fluoroquinolones, and
tetracyclines (Engberg and others 2001;
Rapp 2007)
Antibiotic resistance (de Jong and others
2012)
Prevention:
Vaccine (?)
Control
Reservoirs and risk factors
75.8%, 10-10 000

CFU/g
So, what
about?
21.70%
- 28.11.2007-31.12.2007
- Cross-connection between
sewage- and drinking-water
pipelines
- 9500 residents: 50% ill
- 2052 healthcare visits
- Cost: EUR 354,496: EUR77/
patient
- Regurgitation
- Nausea
- Indigestion
- Heartburn
Sequels
Characteristics of post-campylobacteriosis-diseases.
Adapted from: Smith 2002; Kozminski 2008.
History of Campylobacter
1886: Theodor Escherich: neonates

1913: McFadyean and Stockman: ovine foetuses
1963: Vibrio fetus and Vibrio bubulus Vibrio
1970s:
Filtration technique
Selective media
Source: Charlier and others 1974; Ebruyned and others 2008; Kist 1986; McFadyean and Stockman 1913; Skirrow
1977; Skirrow 2006; Vron and Chatelain 1973
History of Campylobacter
By Joana Revez
15
Campylobacter
Campylobacter coli, Gram-stain, 100X oil immersion

objective
Source: Kaur and others 2011

Really? Another new

Campylobacter species?
Campylobacter iguaniorum (Maarten Gilbert, CDC)

Campylobacter fetus subsp. testudinum (Collette
Fitzgerald, CDC)
Campylobacter stanleyi (Lawson, UK)
New C. laninae-like Campylobacter species (Miller, USDA)
Isolation and detection of

Campylobacter sp.
Challenges:
Low levels of contamination (Oyarzabal and others
2007)
VBNC (Chaisowwong and others 2012).
No Gold standard
Isolation
The ISO 10272-1:2006: Bolton for 4-6 h 37 C

FDA : pre-enrichment 4 h at 37 C or 3 h at 30 C +
2 h at 37 C. Enrichment 42 C for 20-44 h, and
streaking of 24 h and 48 h
The ISO 10272-1:2006: plating in mCCD agar and a 2nd

medium of own choice at 41.5 C for 40-48.
Microaerobic
19 S. M. (2011) The clinical importance of emerging Campylobacter
Man,
species
Nat. Rev. Gastroenterol. Hepatol. doi:10.1038/nrgastro.2011.191. EFSA 2010; EFSA ; ECDC 2013; ISO 2006; ISO 2006
Isolation methods drawbacks

Method of choice impacts the recovery of
Campylobacter from food samples (Edson and
others 2009).
Underestimation of prevalence (Oyarzabal and
others 2013).
Cumulative 9-year false-negative rates (Edson and
others 2009):
C. jejuni : 13.6%
C. coli is 24%
E. coli O157:H7: 7.8%
Salmonella spp. :5.9%
L. monocytogenes : 7.2
Gram-negative
Nonsporeformers
small spiral,
curved, or Sshaped rods (0.2
0.8 m wide and
0.55.0 m long)
Microaerophilic
Common identification methods in

clinical samples
Hip (-) Hip (+)
Who said there is only C. jejuni?
Fitzgerald, CHRO2013
CDC, USA
Presumptive
Campylobacter strains
(864), primarily hippurate
negative strains (20002012)
48% No-C. jejuni/C. coli

24
Typing methods
Adapted from: Moore and others 2006
Typing methods
Typing methods
Phenotype
Grow at 28 C
Hippurate
DNA hydrolysis
H2S production
Resistotyping tests
Penner: soluble
heat-stable
Lior: heat labile
antigens
Penner and Hennessy: 89%, 10 serotypes
Serotyping
Lior and others: 45%, 8 serotypes

Skirrow and Benjamin: Cj: 2 biotypes
60 isolates
Biotyping
Roop and others: Cj: 2 biovars, Cc: 3

Lior: Cj: 2 biotypes, Cc: 2
Preston scheme: Cj: 7 biotypes, Cc: 18
Phage typing
Preston: 3 phage groups

Khakhria and Lior: 3 phage groups
Typing methods
Genotype
MLST
PFGE
fla
70 C. coli: Humans (n=20) and retail meats (n=50)

61 PFGE profiles with SmaI and KpnI
MLST: 37 sequence types
MLST
Source: Sheppard and others 2013
Outbreak: Salinas, Kansas
Outbreak: Walkerton, ON
1346-2300
13
>30
Source: Barton and others 2007; Clark and others 2003
E.coli O157: H7
Virulence factors
fla
cdtB
LOS
WGS
Are non-jejuni/coli emerging

pathogens?
Allarmists
YES
36
Nonbelievers
NO
Why we speak only about C.

jejuni?
It is isolated more
frequently:
Nonbelievers
1. It is the most frequent

enteric pathogen
Allarmists
2. The methods dont

allow the correct
isolation/identification
of other cryptic proteobacteria
37
Underestimation
Undiagnosed human
campylobacteriosis
Unknown etiology:
49% of the 1.7 million cases of foodborne diseases
reported between 1996 and 2000 in England and Wales
(UK)
68% of 41,000 hospital diagnoses of gastroenteritis
reported annually in Australia.
Campylobacter spp. other than C. jejuni/C. coli

frequently be recovered from a substantial portion of
undiagnosed gastroenteritis cases.
Man, S. M. (2011) The clinical importance of emerging Campylobacter species

Nat. Rev. Gastroenterol. Hepatol. doi:10.1038/nrgastro.2011.191
38

Human diarrhoeic samples
Oleastro, CHRO2013
A. cryaerophilus
1%
A. butzleri
5%
Portugal
(from September to
November 2012)
C. jejuni
40%
From 99/299 (33.2%)

stool samples of patients
with diarrhoea positive for
-proteobacteria
Higher in paediatric age
Other Campylobacter spp.

50%
C. fetus
3%

39
C. coli
3%
Bullman, 2011
Cork, Ireland
7194 patient faecal
samples submitted to the
Microbiology
Laboratory. January
2009 and December
2009.
349 (~5%) were

determined to be
Campylobacter
genus-positive

40
Thank you
For the list of references please contact me at: culebroe@mappi.helsinki.fi

Lecture 2 Campylobateria

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Campylobacter

Department of Food Hygiene and Environmental Health

Detection and isolation methods

Confirmed cases per 100 000 population

Source: EFSA 2010; EFSA 2011

Treatment and prevention

Reservoirs and risk factors

75.8%, 10-10 000

1886: Theodor Escherich: neonates

Campylobacter coli, Gram-stain, 100X oil immersion

Source: Kaur and others 2011

Really? Another new

Campylobacter iguaniorum (Maarten Gilbert, CDC)

New C. laninae-like Campylobacter species (Miller, USDA)

Isolation and detection of

The ISO 10272-1:2006: Bolton for 4-6 h 37 C

The ISO 10272-1:2006: plating in mCCD agar and a 2nd

Isolation methods drawbacks

Common identification methods in

Who said there is only C. jejuni?

48% No-C. jejuni/C. coli

Adapted from: Moore and others 2006

Penner and Hennessy: 89%, 10 serotypes

Lior and others: 45%, 8 serotypes

Roop and others: Cj: 2 biovars, Cc: 3

Preston: 3 phage groups

70 C. coli: Humans (n=20) and retail meats (n=50)

Source: Sheppard and others 2013

Outbreak: Salinas, Kansas

Source: Barton and others 2007; Clark and others 2003

Are non-jejuni/coli emerging

Why we speak only about C.

1. It is the most frequent

2. The methods dont

Campylobacter spp. other than C. jejuni/C. coli

Man, S. M. (2011) The clinical importance of emerging Campylobacter species

Who said there is only C. jejuni?

From 99/299 (33.2%)

Other Campylobacter spp.

60% No-C. jejuni/C. coli

Who said there is only C. jejuni?

349 (~5%) were

29% No-C. jejuni/C. coli

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