178 Naturaltoxins: risk, regulations, analysis and ... · EU feed limits exist for aflatoxins EU...

Hans van Egmond, 7 December 2010

Natural toxins:Risks, regulations, analysis

and international co-operation

Natural ToxinsNatural Toxins

mycotoxinsmycotoxins

bacterial toxinsbacterial toxins




phycotoxinsphycotoxins



bacterial toxinsbacterial toxins plant toxinsplant toxins




plant toxinsplant toxins


animal toxinsanimal toxins





� Introduction� Mycotoxins - a continuous issue

� Phycotoxins - interesting developments� Plant toxins - becoming of growing concern

� International cooperation� Summary

Outline of presentation

Mycotoxins:� Naturally occurring chemical compounds produced

by fungi, with adverse effects to man and animals

� Various toxic effects described in man and animals

Toxic effects of mycotoxins

hepatotoxic

immunotoxic

nephrotoxic

neurotoxic

oestrogenic

teratogenic

OTA PAT FUM ZEATCT

carcinogenic

AFL

Mycotoxins:� Naturally occurring chemical compounds produced

by fungi, with adverse effects to man and animals

� Various toxic effects described in man and animals

� Metabolites of mycotoxins also named “mycotoxins”

� Important producing fungi: Aspergillus, Penicillium and Fusarium; hundreds of mycotoxins known

317 people in Kenyan villages got ill; 127 died, including many children

Human aflatoxicosis in Kenya, 2004

Photo: courtesy Henry Njapau and Masja Straetemans

� Village-grown maize suspectedto play a causal role

� FDA emergency team confirmed hypothesis of human aflatoxicosis

� New outbreaks occurred in 2005, 2006 and now again in 2010

Human aflatoxicosis in Kenya, 2004

Photo: courtesy Henry Njapau and Masja Straetemans

Rapid Alert System for Food and Feed

� Quick information-exchange in the EU on risks to human health

� Allows MS to identify potential problems and take measures

� In 2009: 665 mycotoxin issues

RASFF 2009: Border rejection notifications

Border Rejection notifications 2009Rest46%

Veterinary medicine products

4%

Pathogenic micro-

organisms7% Food

additives4%

Compositions3%

Mycotoxins36%

EFSA and JECFA: risk assessments

� JECFA: Joint FAO/WHO Expert Committee on Food Additives

� Risk assessments of variousmycotoxins performed

� Published for AFL, OTA, ZEA,PAT, FUM, DON, T-2/HT-2

� EFSA: risk assessments done for feed, ongoing for food (e.g. various trichothecenes,ergot alkaloids, Alternaria toxins)

� Worldwide inquiries:1981, 1987, 1995 & 2003, resulting in various publications

� Inquiry 2003 published as FAO FNP 81 (2004)

� Chinese, French and Spanish translations available

Inquiries on mycotoxin regulations

Mycotoxin regulatory situation in Europe

� 39 nations with known regulations � EU food limits exist for AFLAs,

OTA, PAT, ZEA, DON and FUM� EU food limits discussed for

T-2/HT-2 toxins and ergot alkaloids� The near future: Alternaria toxins

and several other mycotoxins� EU feed limits exist for aflatoxins� EU feed guidance limits established

for OTA and Fusarium mycotoxins

Analytical methodology for mycotoxins

� US based, international involvement

� Development and validation of methods of analysis and improvement of AQA

� Approx. 45 mycotoxin methods in “OMA”

� European equivalent of ISO� Performance criteria approach, usually

based on interlaboratory studies� 12 mycotoxin methods standardized� EU interlaboratory studied methods CEN

AOAC and CEN mycotoxin methods

� TLC: veteran technique, powerful and cheap� HPLC: frequently used and extensively

validated, often in combination with IA cleanup

� ELISA: especially useful for rapid screening

Newer developments in methodology

� LC-MS and GC-MS� Biosensors� Capillary Zone Electrophoresis� Infrared techniques� Acoustic methods� Molecular Imprinted Polymers� Fluorescence Polarisation Immunoassay� Dip Stick Immunoassay� Transcriptomics

“Red tides” - explosive growth of algae

Dinoflagellates, small toxin-producers

Dinophysis acuminata

Pyrodinium bahamense

Groups of marine biotoxins

� “Paralytic shellfish poisoning” toxins (PSP)� “Diarrhoeic shellfish poisoning” toxins (DSP)

� Azaspiracids (AZA)� “Amnesic shellfish poisoning” toxins (ASP)� Emerging: palytoxins,

brevetoxins, cyclic imines

� Ciguatera toxins

The EU situation on phycotoxins

� Several toxin groups regulated(OA, AZA, YTX, PTX, STX, DA)

� Rodent assays will soon be banned

� EFSA suggests more stringent limits for various regulated toxins; emerging toxins discussed

� Interesting chemical alternatives to MBA explored

� Networks exist (CEN, CRL/NRL), and someprojects: BioCop, MoniQA, CONffIDENCE

Consequences of climate change

� Expected increase of algal blooms � More, more frequent and more types of marine

biotoxins in fishery products (“emerging toxins”)� Example: ciguatera toxins now found in

Madeira and the Canary islands (Europe)� Governments to take adequate measures for

risk management� Governments to be encouraged to carry out

integral monitoring (algae and shellfish) to protect the consumer

Research needs

� Improve systems to predict harmful algalblooms for risk management

� Prediction of algal blooms: knowledgerequired about modelling and ecology

� Shift research from local to global scale tobetter understand the impact of climate change

� The increased chance of occurrence ofmarine biotoxins requires more knowledgeof toxicology and analytical possibilities

Plant toxins: growing concern

� Recurring incidents, sometimes very serious

� Regulations for food and feed scarce� Increased interest in the EU, e.g. for alkaloids� EFSA opinions: focus on validation of analytical

methods, development of RMs and surveys � Data to be produced on carry-over � Limited project activities exist (CONffIDENCE)

Outbreak of liver disease in Hirat district

Outbreak of liver disease in Hirat district

� More than 270 people affected

� Approx. 50 people died

� WHO alarmed, and RIVMcontracted to investigatethe cause of the disease

� Hypothesis: PA poisoning

� Samples sent to RIVM

� PA poisoning confirmed

Pilot carry-over study ragwort

Examples of European collaboration

� Many networks and activities ongoing

� Rapid Alert System for Food and Feed

� EFSA: scientific opinions (risk assessments)

� CEN: the European equivalent of ISO

� EU-RLs for mycotoxins and marine biotoxins

� European research and networking projects

NRL networks on mycotoxins and marine biotoxins

27 NRLs

1 EU-RL

>> RFLs

EC

Tasks of the EU-RL

27 NRLs

1 EU-RL

>> RFLs

EC

� Provide NRLs with methods

� Coordinate comparative testing

� Organise training courses

� Technical assistance to the Commission

� Collaborate with labs in third countries

Examples of European collaboration

� Many networks and activities ongoing

� Rapid Alert System for Food and Feed

� EFSA: scientific opinions (risk assessments)

� CEN: the European equivalent of ISO

� EU-RLs for mycotoxins and marine biotoxins

� European research and networking projects

� IP Food Quality & Safety, 2005-2010 (just ended)

� Biotech-based methods explored and exploited� Includes WPs on mycotoxins and phycotoxins

� Involves various analytical techniques(such as sensors, transcriptomics)

� Continuation in national projects & interaction with MoniQA is foreseen (validation studies)

MoniQAMoniQA

� NoE Food Quality & Safety, 2007-2012

� Focus on validation of and performance criteriafor methods to monitor food hazards

� Includes activities on different groups of analytes

� Emphasis on rapid and high throughput methods� Active WG on mycotoxins/phycotoxins

� Mycotoxin workshops and interlaboratory studies form part of the activities

� LCP Food, Agriculture and Fisheries, and Biotechnology, 2008-2012

� Simple, fast, multi-analyte, multi-class detection � Includes WP Biotoxins

- alkaloids (pyrrolizidine, tropane, ergot)- Fusarium mycotoxins (TCT, ZEA, FUM)- Various types of phycotoxins (PSP, DSP, ASP)

� Training workshops and educating modules� Interaction with MoniQA (validation studies)

� LCP Food, Agriculture and Fisheries, and Biotechnology, 2009-2013

� “Reduction of mycotoxins in food and feed chains”� Focus on AFL, OTA, TCT, ZEA en FUM � Rapid methods (strips, LFD, multimycotoxin tests)� Analytical WP: service to other work packages � Organisation of (training) workshops and

conferences (Europe, Asia, Africa, Latin America)

Summary and conclusions

� Natural toxins: a worldwide concern� Mycotoxins: many regulations, analytical

situation advanced, strong networks� Phycotoxins: regulations need revision,

analytical progress, networks growing � Plant toxins: regulations scarce, analytically

least advanced, networks to be formed� Climate change will affect natural toxins� Several international activities ongoing

to cope with the problems of natural toxins

Thank you for listening !

Natural toxins:Risks, regulations, analysis

and international co-operation

Consequences of climate change

� Expected changes in range of latitudes, where certain fungi are able to compete

� Example: F. graminearum growth: NIV increase� Drought, flooding and other consequences of

climate change may result in more mycotoxinsand changed toxin profiles

� Example: aflatoxins found in Italy since 2003,and in other parts of Central Europe

� Response of insects and plant diseases to climate change poorly understood, but increases expected

Research needs

� Better understanding of precise impact of climate change on insects and other pests

� Investigate consequences on soil characteristics: very relevant for fungal infestation

� Stimulate research from local to global scale tobetter understand the impact of climate change

� EMTOX project: one of the objectives: Quantify relationship between climate change and toxins

� Monitoring of mycotoxin occurrence, to observe possible changes in toxin profiles and levels

Various translations of FNP 81 available

� Succesful carry-over experiment� Various PA found in milk, jacoline major

compound (transfer: 6-8 % of ingested amount)� Risk assessment by RIVM: allowed daily weed

intake of cow: 0.66 g/day� EU - workshop on PA held, February 2010� Second carry-over study ongoing� EFSA: opinions requested for various alkaloids

and possibly other plant toxins in food

Early conclusions and follow-up

178 Naturaltoxins: risk, regulations, analysis and ... · EU feed limits exist for aflatoxins EU...

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