sampling passengers foods at airports: do we carry ... wagner.pdflisteria monocytogenes mlst typing...

Funded by the 7th Framework

Programme of the European

Union

FP7 – Knowledge-based Bio-Economy

(KBBE)

All States Veterinarian Conference, Limerick, 16th to 18th of April, 2015

Martin Wagner

Professor for Molecular Food Microbiology, Head of Institute for Milk

Hygiene and Food Science, Department for Farm Animals and

Veterinary Public Health, University for Veterinary Medicine, Vienna

Sampling passengers foods at airports: Do

we carry pathogens in our hand luggage?



Union


(KBBE)

David Rodriguez-Lazaro,

Jordi Rovira, and team

Bela Nagy

and team

Janine Beutlich,

Bernd Appel

Sonja Smole-Mozina

and team

Acknowledgement

Luminita Cirolacu

Anca Nicolau



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(KBBE)

Lunch for two : www.maps.ethz.ch

Shrimps (Thailand)

Onions (New Zealand)

Ginger (China)

Avocado (Israel)

Chili (Argentina)

Pecan nut (Gabun)

Peperoni (Spain)

Origin of food items used to produce a

shrimp wrap



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(KBBE)

PROMISE (7th FP program)-WP1:

macrotransmission of foodborne pathogens

Studied contamination rate of food items carried by global travellers (airports

Vienna n=600; Frankfurt/Schönefeld n=823; Bilbao n=200, Slovenija n=66)

Studied contamination rate of food items traded by small black

markets in Romania (n=200)

Studied contamination rate of food items traded at market selling

mainly homegrown food in Turkey (n=200)

Studied contamination rate of food items carried away across

ground borders and seaports located in South-West Europe

Studied contamination rate of food items imported to Greece

• All airports: Salmonella spp., L. monocytogenes, Campylobacter, VTEC

• Vienna: E. coli, Staphylococci

• Burgos: Food-borne viruses, Staphylococci



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(KBBE)

Procedures

Isolations done in partner labs

Salmonella

spp.

Listeria

monocytogenes

MLST typing

Virulotyping

Escherichia

coli

Campylobacter

jejuni/ coli

Staphylococcus

aureus

Micro array

MDR Verotoxin MDR Virulotyping MDR

Sample number/type of food to be tested approved

Harmonisation of methods



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(KBBE)

14.777.196 (8-fold of the Viennese population)

78.415

58.795

• 61355 travellers controlled

• 1473 products of animal origin were confiscated

• The food confiscates totalled to an amount of 6229 kg

• 600 samples tested for pathogens and hygiene indicators

Confiscate sampling Vienna

(August 2012 – March 2013)



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(KBBE)

Experiences from Vienna airport:

identifiable food items

Cairo, Egypt Seoul, south Korea

Yerevan, Armenia St. Petersurg, Russia Delhi, India



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(KBBE)

Experiences from Vienna and Bilbao

airport: exotic food items

Duck tongues, China Egg (1000 years old)

Rat (courtesy UoB)

Squirrel (courtesy UoB)



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(KBBE)

Experiences from Vienna airport:

unidentifiable origin

Bush meat, Nigeria Bush meat, Ethiopia

Kudu biltong, South Africa



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(KBBE)

48.2% of samples orginated from 33 other countries worldwide

51.8% of samples orginated from Turkey!

0 5 10 15 20 25 30 35

Africa

Asia

Third Country (Europe)

South America

Where did the food commodities

originate from?

China n=80

Egypt n=65



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(KBBE)

96.2% of samples: meat and milk products

Milk&products

n=315 7 5 Bush meat

n=6

83.2% (hard cheese)

11.4% (semihard)

5.4% (fresh)

47% (cooked meat)

17.2% (raw meat)

35.8% (sausages)

Meat&products

n=262

Fish&products

n=6



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(KBBE)

38.7% 61.3% 23.5% 44.8%

11.5% 4.3% 4.3% 3.8%

3.0% 1.7% 1.3% 0.3%

Types of packaging



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(KBBE)

Summary

Turkey

(n=311)

China

(n=80)

Africa

(n=85)

Others

(n=128)

Salmonella spp. 1 0 4,7 0

L. mono. 3,5 0 0 1,5

VTEC 2,6 0 0 0

SA 3,2 0 5,8 1,5

E. coli 13,2 0 21,2 5,4 0

2

4

6

8

10

12

Pathogens Indicators

Prevalence Samples Vienna Prevalence regional



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(KBBE)

ST-types matched with origin of food items

48 L. monocytogenes strains were subjected to MLST typing according to

Ragoon et al. (2008) 17 different ST-types (7 Lineage I, III; 100 Lineage II)



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(KBBE)

Data Listeria monocytogenes

Comparison of 48 L.

monocytogenes strains to all ST

types stored to the Pasteur st

database 2010-2014 (n=191)



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(KBBE)

Are the ST-types found fully virulent?

EGDe RO01

RO02 RO03 RO04

RO05

RO06

RO07

RO08

RO09

RO10 RO11

RO12

RO13

RO14

RO15

EGDe RO01 RO02 RO03

RO04

RO05

RO06

RO07

RO08

RO09 RO10

RO11

RO12

RO13

RO14

RO15

-5

0

5

10

15

20

25

30

0 1 2 3 4 5 6 7 8

Intr

ace

lllu

lar

Gro

wth

Co

efic

ien

t

Invasion efficiency, %

THP-1

Caco-2

ST 155

ST121

ST20



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(KBBE)

Are ST-types found fully virulent?

Only ST-types 2 and 20 fully virulent



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(KBBE)

10 5 10 5

Low virulent strains, sequencing of inlA gene

Number of mutations

Function E-cadherin binding Cell wall anchor Structural



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(KBBE)

inlA mutations

LRR domain Ig-like Transmembrane Anchor domain



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(KBBE)

….If ST121 strains are so prevalent (even if recovered

from food originated different countries and continents)

what increases their survival capacity?

The crucial question…..

ST121 strains (of low relevance for public health) were

most frquently recovered……



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(KBBE)

Sequences of ST121 strains isolated from

different countries are highly conserved



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(KBBE)

All sequenced ST121 strains show deletions in the

inlA gene (adapted in virulence)!

EGDe

1 kbp

6179 inlA* lmo0435/bapL*

4423 inlA* lmo0435/bapL*

N53-1 inlA* lmo0435/bapL*

LM_1880 inlA* lmo0435/bapL*



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(KBBE)

The ST121 plasmids are almost identical



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(KBBE)

Prophage 6179 tRNA-Arg-TCT

Pro

ph

ag

e 4

42

3 t

RN

A-A

rg-T

CT

Prophage 6179 tRNA-Arg-CCG

Pro

ph

ag

e 4

42

3 t

RN

A-A

rg-C

CG

tRNA-Arg-TCT and tRNA-Arg CCG prophages

are highly conserved among ST121 strains

Prophage 6179 tRNA-Arg-TCT

Pro

ph

ag

e L

. L

M_

18

80

tR

NA

-Arg

-TC

T

Prophage 6179 tRNA-Arg-CCG

Pro

ph

ag

e L

M_

18

80

tR

NA

-Arg

-CC

G

Prophages in N53-1 and LM_1880 are in multiple contigs



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(KBBE)

RHS protein

6179

4423

RHS protein

N53-1

RHS protein

The ST121 genomes contain an insertion possibly

involved in competition against other bacteria



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(KBBE)

Putative function of the RHS proteins

• Mediate competition by inhibiting growth of neighboring

cells

• In ST121 strains: KptA upstream:

– RNA 2‘ phosphotransferase: Catalyzes RNA cleavage (most

likely also tRNA cleavage)



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(KBBE)

Conclusions: Traveller behaviour

• 2,4%-4.2% of travellers carry food in their luggage

• Amount of food carried by travellers is enormous (at an

average 4,2kg; 14kg of raw poultry meat in one case)

• Only few nationalities contribute to the majority of food

commodities shuttled (immigrant`s scene; Case airport

Vienna: Turkey, Egypt, China app. 77%)

• In many cases, travellers carrying food are not the endusers

• Due to limited distribution, the public health impact is likely

low ( „Restaurant outbreaks“)

• Travellers are not aware of legal restrictions



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(KBBE)

Conclusions: Hazard

characterization

• Hygiene of this products was inacceptable in most cases

• Some food confiscated from travellers were contaminated at

rates higher than described in samples taken from domestic

production

• No classical foodborne viruses detected, however HepE

highly prevalent in samples confiscated at airport Bilbao

• Campylobacter was found in neither samples (so as in

samples from airports Bilbao and Frankfurt and Schönefeld)

• Strain characterization: no superbugs detected until now,

however new types were found!

• The crucial question regarding L. monocytogenes: Do we

need to perform risk assessments on subtype level??



Union


(KBBE)

www.promise-net.eu

Special issue on Microbial Transmission appearing

in International Journal of Food Microbiology soon

sampling passengers foods at airports: do we carry ... wagner.pdflisteria monocytogenes mlst typing...

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