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Worldwide and European regulationsfor mycotoxins & analytical methods

expected from Biocop

Hans van Egmond & Marco Jonker, Brussels, 22 October 2004

RENational Institutefor Public Health

and the EnvironmentResearch for man and environment

2Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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“It can be measured, so it must exist”

Quote of first speaker at conference: “Measurements, a key to competitiveness”, 25th anniversary of BCR, November 1998

3Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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“A well-known mycotoxin, such as aflatoxin B1 is 33 million times more dangerousthan the dangerous solvent carbon tetrachloride – itsthreshold is 0.000001 gramper kg per day”

Naresh Magan, RTD Info 42, August 2004

4Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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“There is no point imposingconstraining regulations ifonly a few individuals and laboratories are able to monitor their applicationthroughout the continent”

Naresh Magan, RTD Info 42, August 2004

5Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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Outline of presentation

• Introduction• Factors influencing mycotoxin regulations• International inquiries on mycotoxin regulations• “Tour-du-monde“ 2003 & focus on the EU• Worldwide limits for aflatoxins in food and feed• Worldwide limits for other mycotoxins in food• BioCop work package mycotoxins• Summary

6Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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Outline of presentation

• Introduction• Factors influencing mycotoxin regulations• International inquiries on mycotoxin regulations• “Tour-du-monde“ 2003 & focus on the EU• Worldwide limits for aflatoxins in food and feed• Worldwide limits for other mycotoxins in food• BioCop work package mycotoxins• Summary

7Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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Factors influencing mycotoxin regulations

• Availability of toxicity data• Availability of survey analytical data• Availability of methods of sampling and analysis• Trade contacts with other countries• Sufficiency of food supply

8Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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Weighing the various factors: not trivial

9Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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Outline of presentation

• Introduction• Factors influencing mycotoxin regulations• International inquiries on mycotoxin regulations• “Tour-du-monde“ 2003 & focus on the EU• Worldwide limits for aflatoxins in food and feed• Worldwide limits for other mycotoxins in food• BioCop work package mycotoxins• Summary

10Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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Inquiries on mycotoxin regulations

• Worldwide inquiries in 1981, 1987, 1995 and 2002/2003, resulting in various publications

• Inquiry 1995 published as FAO FNP Paper 64 (1997)

11Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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International inquiry 2002/2003

Questionnaire February 2002For an Update of

“Worldwide regulations for mycotoxins 1995”FAO Food and Nutrition Paper 64

• Inquiry 2002/2003: FAO-contracted activity of RIVM• Information: Dutch Embassies and personal contacts • Details asked a.o. about tolerance limits, legal bases,

responsible authorities, methods of sampling and analysis• Results: regulations exist in 100 countries and for 13 toxins• FAO Food and Nutrition Paper: in print

12Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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Outline of presentation

• Introduction• Factors influencing mycotoxin regulations• International inquiries on mycotoxin regulations• “Tour-du-monde“ 2003 & focus on the EU• Worldwide limits for aflatoxins in food and feed• Worldwide limits for other mycotoxins in food• BioCop work package mycotoxins• Summary

13Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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Africa

• 15 nations with known regulations (59 % of inhabitants of the region)

• Majority of countries: regulations unknown or non-existent

• Several countries indicate regulations should be developed

• Regulations mainly for AFB1

• Most detailed: Morocco

14Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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Asia/Oceania

• 26 nations with known regulations (89 % of inhabitants of the region)

• Regulations for total aflatoxinsdominate in food, regulations for AFB1 dominate in feed

• Harmonized regulations in Australia & New Zealand

• Most detailed : China and Iran

15Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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Latin America

• 19 nations with known regulations (92 % of inhabitants of the region)

• Harmonized aflatoxin regulations in MERCOSUR

• Aflatoxin regulations mostly set for total aflatoxins

• Most detailed regulations: Uruguay

16Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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North America

• 2 nations with known regulations (100 % of inhabitants of the region)

• Canada: detailed tolerances forFusarium damaged kernels (% FDK) and for ergot (% by weight)

• USA: detailed tolerances forfumonisins (B1+B2+B3) in a widevariety of foods and feeds

17Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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Europe

• 39 nations with known regulations 99 % of inhabitants of the region)

• EU harmonized limits exist for: - total aflatoxins and AFB1 in various

foodstuffs and AFB1 in baby food- AFM1 in milk and baby food- ochratoxin A in various

foodstuffs and baby food- patulin in fruit products and baby food

• Low limits applied, especially for baby food

18Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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Europe - regulations in development

• EU guidelines limits for DON will be “upgraded” to regulated max. levels

• Expected: EU limits for ochratoxin A in coffee, wine and grape juice

• Expected: EU regulations for several Fusarium toxins in food and baby food

• Expected: EU regulations for various mycotoxins in feeds (apart from AFB1)- DON, zearalenone, fumonisins - ochratoxin A - ergot alkaloids

19Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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Europe: mycotoxins regulated in food (2003)

0 5 10 15 20 25 30 35 40

AFT

AFB1

AFB1/G1

AFM1

OTA

PAT

DON

T-2

ZEN

FUMB1/2

FUMB1

STEm

yco

toxi

ns

number of countries

20Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

r

Europe: mycotoxins regulated in feed (2003)

0 5 10 15 20 25 30 35 40

AFT

AFB1

AFB1/G1

OTA

DON

T-2

DAS

ZEN

myc

oto

xin

s

number of countries

21Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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AFRICA

EUROPE

ASIA

&

OCEANIA

NORTH AMERICA

LATIN AMERICA

22Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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Number of mycotoxin regulations per country

Food

1,53,6

1,8

5,519,6

ASIA / OCEANIA

LATIN AMERICA

AFRICA

EUROPE

NORTH AMERICA

23Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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Number of mycotoxin regulations per country

Feed1,3

1,71,0

13,5

8,5

EUROPE

NORTH AMERICA

LATIN AMERICA

ASIA / OCEANIAAFRICA

24Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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Outline of presentation

• Introduction• Factors influencing mycotoxin regulations• International inquiries on mycotoxin regulations• “Tour-du-monde“ 2003 & focus on the EU• Worldwide limits for aflatoxins in food and feed• Worldwide limits for other mycotoxins in food• BioCop work package mycotoxins• Summary

25Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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Aflatoxin B1 in food

1

29

21

5

2

3

1 µg/kg

2 µg/kg

5 µg/kg

10 µg/kg

15 µg/kg

20 µg/kg

number of countries

26Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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Total aflatoxins in food

23

129

388

173

2

0 µg/kg

1 µg/kg

3 µg/kg

4 µg/kg

5 µg/kg

10 µg/kg

15 µg/kg

20 µg/kg

30 µg/kg

35 µg/kg

number of countries

27Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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Aflatoxin M1 in milk

1

34

1

22

1

1

not detectable

0.05 µg/kg

0.2 µg/kg

0.5 µg/kg

5 µg/kg

15 µg/kg

number of countries

28Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

r

Aflatoxin B1 in feed for dairy cattle

27

5

1

3

1

2

5 µg/kg

10 µg/kg

15 µg/kg

20 µg/kg

25 µg/kg

50 µg/kg

number of countries

29Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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Outline of presentation

• Introduction• Factors influencing mycotoxin regulations• International inquiries on mycotoxin regulations• “Tour-du-monde“ 2003 & focus on the EU• Worldwide limits for aflatoxins in food and feed• Worldwide limits for other mycotoxins in food• BioCop work package mycotoxins• Summary

30Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

r

Patulin in fruits and fruit juices

1

1

1

44

1

5 µg/kg

25 µg/kg

30 µg/kg

50 µg/kg

100 µg/kg

number of countries

31Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

r

Ochratoxin A in cereals and cereal products

1

29

1

1

1

1

3

3 µg/kg

5 µg/kg

10 µg/kg

15 µg/kg

20 µg/kg

30 µg/kg

50 µg/kg

number of countries

32Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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DON in wheat(flour) and other cereals

1

4

19

9

1

2

1

300 µg/kg

700 µg/kg

750 µg/kg

1000 µg/kg

1100 µg/kg

1200 µg/kg

2000 µg/kg

number of countries

33Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

r

Zearalenone in maize and other cereals

1

1

2

5

8

50 µg/kg

60 µg/kg

100 µg/kg

200 µg/kg

1000 µg/kg

number of countries

34Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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Fumonisins in maize

4

1

1

1000 µg/kg

2000 µg/kg

3000 µg/kg

number of countries

35Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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Techniques used in mycotoxin regulatory analysis

TLC

minicolumn

LC

GC

GC/MS ELISA

fluorometry

frequency of techniques

36Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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Outline of presentation

• Introduction• Factors influencing mycotoxin regulations• International inquiries on mycotoxin regulations• “Tour-du-monde“ 2003 & focus on the EU• Limits for aflatoxins in food and feed• Limits for other mycotoxins in food• BioCop work package mycotoxins• Summary

37Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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An Integrated Project proposal submitted to the EC on the subject of chemical contaminant monitoring

38Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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Broad Spectrum Contaminant Analysis

In their wise & far sighted approach to the subject the European Commission published in the FP6 Food Quality Work Programme:

“New methods to prevent & monitor occurrence of multiple contaminants such as pesticides, toxins, drugs and endocrine disrupters in foods.

This should be achieved through use of advanced sample preparation techniques & emerging biotechnological screening approaches based on development of novel biomarkers.”

39Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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Question…….

How can such a radical set of requirements be met?

The birth of the consortium (32 partners)

The BioCop principles:The fundamental scientific objective is to utilise emerging lifescience technologies to create a powerful new approach to detect & control chemical contaminants in foods.

40Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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BioCop: the science…..

Veterinary drugs, heavy metals, mycotoxins, endocrine disrupters,

shellfish toxins, pesticides

Transcriptomics, proteomics, biosensors, molecular engineering

41Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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The BioCop matrixWP1 WP2 WP3 WP4

Binding moleculesTranscriptomics Proteomics Biosensors

WP5 Shellfish toxins

WP10 Mycotoxins

WP6 Pesticides

WP8 Therapeutic drugs

WP7 Endocrine disrupters

WP9 Heavy metals

42Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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Work package mycotoxins

• 8 partners involved • Links with CEN and AOAC International• Focus on trichothecenes (DON, NIV, T-2, HT-2)• Techniques explored:

- new analyte isolation techniques (e.g. pressurized liquid extraction, slurrying techniques, microwave hydrolysis)

- transcriptomics (identification of chemical residues through genomic fingerprints)

- electrochemical biosensors (low cost portable devices)

43Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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Phase 1: Research, e.g. Development of binders, surface chemistry, chip production, equipment design, test materials

Phase 2: Research, e.g. Development of assays, technology coupling, sample preparation

Phase 3: Comparison and validationCompare new assays with state of the art & fully validate

Phase 4: Demonstration Technology transfer events, e.g. workshops

YEAR

1

2

3

4

5

Decision Points Decision Points

Decision Points Decision Points

Decision Points Decision Points

BioCop phases

44Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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Current status of the BioCop proposal

Submitted February 2004

First phase of evaluation: Passed

Second phase evaluation: Passed

Contract negotiations ongoing since June 2004

Completion of contracts: November 2004

Five year project from January 2005, 32 partners from 16 countries, EC contribution approx. 10M €

45Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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Outline of presentation

• Introduction• Factors influencing mycotoxin regulations• International inquiries on mycotoxin regulations• “Tour-du-monde“ 2003 & focus on the EU• Limits for aflatoxins in food and feed• Limits for other mycotoxins in food• BioCop work package mycotoxins• Summary

46Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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Summary

• Various factors involved in establishing mycotoxin regulations • Mycotoxin regulations exist in 100 countries and for 13 toxins• Regulations in 2003: more diverse and detailed than in 1995• Harmonisation takes place, yet regulatory limits remain

substantially different across countries• Details of 2002/2003 inquiry: FAO FNP in 2004• BioCop: new technologies (transcriptomics, biosensors)

explored for determination of trichothecenes

Thank you for your attention!

Worldwide and European regulationsfor mycotoxins & analytical methods

expected from Biocop

Hans van Egmond & Marco Jonker, Brussels, 22 October 2004

RENational Institutefor Public Health

and the EnvironmentResearch for man and environment

48Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

r

49Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

r

Africa: mycotoxins regulated in food

0 2 4 6 8 10 12

AFT

AFB1

AFM1

AFG1

OTA

PAT

ZENm

ycot

oxin

s

number of countries

50Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

r

Africa: mycotoxins regulated in feed

0 2 4 6 8 10 12

AFT

AFB1

AFB1/G1

OTA

ZEN

myc

otox

ins

number of countries

51Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

r

Asia/Oceania: mycotoxins regulated in food

0 5 10 15 20

AFT

AFB1

AFM1

OTA

PAT

DON

T-2

ZEN

FUMB1/2

FUMB1

AGA

PHOm

yco

toxi

ns

number of countries

52Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

r

Asia/Oceania: mycotoxins regulated in feed

0 5 10 15 20

AFT

AFB1

OTA

DON

T-2

DAS

ZEN

myc

oto

xin

s

number of countries

53Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

r

Latin America: mycotoxins regulated in food

0 5 10 15 20

AFT

AFB1

AFB1/G1

AFM1

OTA

PAT

DON

ZEN

FUMB1m

yco

toxi

ns

number of countries

54Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

r

Latin America: mycotoxins regulated in feed

0 5 10 15 20

AFT

AFB1

DON

ZEN

ERG

myc

oto

xin

s

number of countries

55Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

r

North America: mycotoxins regulated in food

0 2 4 6 8 10

AFT

AFM1

PAT

DON

FUMB1/2/3

myc

oto

xin

s

number of countries

56Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

r

North America: mycotoxins regulated in feed

0 2 4 6 8 10

AFT

DAS

DON

ERG

FUMB1/2/3

HT-2

OTA

T-2

ZEN

myc

oto

xin

s

number of countries

57Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

r

Information in draft table of FAO document

status ref. status ref

FOODGroundnuts, nuts, dried fruit

Aflatoxin B1 2 [EU 1] Min. Health official [EU 2] official [EU 2]

DAIRYMilk Aflatoxin M1 0.05 [EU 1] Min. Health official [EU 2] official [EU 2]

FEEDgrain (products) incorporated in feed for pigs

DON 5000 [NL 1] Commodity Board Animal

Feedstuffs

Sampling method

Analytical method

The Netherlands <NL> 2002:

Legal basis

Responsible authority

Commodity Mycotoxin Limit (ng/g)

58Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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AFRICA

EUROPE

ASIA

&

OCEANIA

NORTH AMERICA

LATIN AMERICA

59Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

r

60Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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Number of mycotoxin regulations per country

Dairy

0,3

0,7

0,60,5

1,2

AFRICA

ASIA / OCEANIA

LATIN AMERICANORTH AMERICA

EUROPE

61Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

r

Total aflatoxins in feed for dairy cattle

2

4

7

3

5

zero/0.01 µg/kg

10 µg/kg

20 µg/kg

30 µg/kg

50 µg/kg

number of countries

62Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

r

Ranges of limits for total aflatoxins in food

0 5 10 15 20 25 30 35 40 45 50

µg/kg

2002

North America 1995

2002

Latin America 1995

2002

Europe 1995

2002

Asia/Oceania 1995

2002

Africa 1995

63Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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Typical limits for total aflatoxins in food

0 5 10 15 20 25 30 35 40 45 50

µg/kg

2002

North America 1995

2002

Latin America 1995

2002

Europe 1995

2002

Asia/Oceania 1995

2002

Africa 1995

64Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

r

65Mycotoxin regulations & BioCop / Van Egmond & Jonker, Brussels 2004

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The BioCop matrixWP1 WP2 WP3 WP4

Binding moleculesTranscriptomics Proteomics Biosensors

WP5 Shellfish toxins

WP10 Mycotoxins

WP6 Pesticides

WP8 Therapeutic drugs

WP7 Endocrine disrupters

WP9 Heavy metals

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