Lecture 2 Campylobateria

www.helsinki.fi/yliopisto www.helsinki.fi/yliopisto

Campylobacter

Alejandra Culebro

Department of Food Hygiene and Environmental Health

Faculty of Veterinary Medicine

www.helsinki.fi/yliopisto

Campylobacteriosis

The genus Campylobacter

History

Taxonomy

Detection and isolation methods

Typing methods

Emerging campylobacters

Overview

www.helsinki.fi/yliopisto 30.10.2013 3 Source: National Institute for Health and Welfare 2011

0

10

20

30

40

50

60

70

80

90

2007 2008 2009 2010 2011

Co

nfi

rmed

cases p

er

100 0

00 p

op

ula

tio

n

Year

Average of EU member states except for Latvia, Portugal,Romania, and Slovenia

Finland

Source: EFSA 2009; EFSA 2010; EFSA 2011; EFSA and ECDC 2012; EFSA and ECDC 2013

Salmonella Campylobacter


Campylobacter

Source: THL

www.helsinki.fi/yliopisto Source: EFSA 2010; EFSA 2011

www.helsinki.fi/yliopisto 6

0

20

40

60

80

100

120

140

160

180

200

Fin EU US CR NewZealand

UK


500-800 cells

3.2 days

Self-limiting

Relapse 5-10%

Symptoms

Source: Black and others 1988; Feodoroff and others 2011; Pacanowski 2008; Skirrow 1977

Yet - Ingested dose

- Strains virulence - Acquired immunity

- Health status


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


So, what

about?

21.70%

75.8%, 10-10 000

CFU/g


- 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.


1886: Theodor Escherich: neonates

1913: McFadyean and Stockman: ovine foetuses

1963: Vibrio fetus and Vibrio bubulus Vibrio

1970s:

Filtration technique

Selective media

History of Campylobacter

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


Campylobacter

Source: Kaur and others 2011

Campylobacter coli, Gram-stain, 100X oil immersion

objective


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)


Challenges:

Low levels of contamination (Oyarzabal and others 2007)

VBNC (Chaisowwong and others 2012).

No Gold standard

Isolation and detection of

Campylobacter sp.


19

Isolation

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

Nat. Rev. Gastroenterol. Hepatol. doi:10.1038/nrgastro.2011.191. EFSA 2010; EFSA ; ECDC 2013; ISO 2006; ISO 2006

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


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 S-

shaped rods (0.20.8 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 (2000-

2012)

48% No-C. jejuni/C. coli


Typing methods

Adapted from: Moore and others 2006


Typing methods


Typing methods

Phenotype

Penner: soluble

heat-stable

Lior: heat labile

antigens

Grow at 28 C

Hippurate

DNA hydrolysis

H2S production

Resistotyping tests


60 isolates

Serotyping

Penner and Hennessy: 89%, 10 serotypes

Lior and others: 45%, 8 serotypes

Biotyping

Skirrow and Benjamin: Cj: 2 biotypes

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

E.coli O157: H7

>30 Source: Barton and others 2007; Clark and others 2003


Virulence factors

fla

cdtB

LOS


WGS


Are non-jejuni/coli emerging

pathogens?

36

Allarmists Nonbelievers

YES NO


It is isolated more frequently:

1. It is the most frequent

enteric pathogen

2. The methods dont allow the correct

isolation/identification

of other cryptic -proteobacteria

Why we speak only about C.

jejuni?

37

Underestimation

Allarmists

Nonbelievers


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.

38

Undiagnosed human

campylobacteriosis

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

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


Human diarrhoeic samples

C. jejuni

40%

Other Campylobacter spp.

50%

C. coli

3% C. fetus

3%

A. cryaerophilus

1%

A. butzleri

5%

39


Oleastro, CHRO2013

Portugal

(from September to

November 2012)

From 99/299 (33.2%)

stool samples of patients

with diarrhoea positive for

-proteobacteria

60% No-C. jejuni/C. coli

Higher in paediatric age



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 29% No-C. jejuni/C. coli

www.helsinki.fi/yliopisto www.helsinki.fi/yliopisto

Thank you

For the list of references please contact me at: [email protected]

Documents

Lecture 2 Campylobateria