United Kingdom Anisakis Reference Laboratory: Code of Practice/Guidance Document

United Kingdom Anisakis Reference Laboratory: Code of Practice/Guidance Document

(DOC32 - Revision 1)

Centre for Environment, Fisheries and Aquaculture Science,

Weymouth Laboratory, Weymouth, Dorset, UK, DT4 8UB

Tel [phone], Fax : [phone].

1.Version Control

Submitted to:	Steve Feist
Date submitted	18/3/2015
Project manager	Steve Feist
Document compiled by:	Tom Hill
Quality control by:	Steve Feist
Approved by and date:	Steve Feist 18/3/2015
Version:	1.3

Author	Date	Comment	Version
Tom Hill	6/3/2015	DRAFT	1.1
Steve Feist	18/3/2015	Minor changes to document	1.2
Tom Hill	19/3/2015	Saved to contracts drive	1.3
Tom Hill	25/6/2015	Version control added and uploaded to QPulse as ‘xDoc3- Revision 1’	xDoc3 -Revision 1
Tom Hill	2/7/2015	Error with naming convention. Changed to DOC32.	DOC32 – Revision 1

2.Preface

This guidance document provides a brief code of practice for the monitoring of zoonotic parasites from the family Anisakidae within fish produce destined for human consumption. It provides information relative to: the National Reference Laboratory of the United Kingdom; the European Reference Laboratory; the Official Control Laboratories which are associated with the laboratory network overseen by the European Reference Laboratory; industry; and consumers.

The majority of the legislative information contained within is referenced from European Union ‘Regulation (EC) No 882/2004’ and ‘Regulation (EC) No 853/2004’ and is provided for context and information purposes. For in depth understanding of the legislation, it is highly recommended that stakeholders consult the source documents.


Table of Contents

1.Version Control 3

2.Preface 4

3.Introduction 6

4.Administration 7

3.1Introduction 7

3.2The European Union Reference Laboratory (EURL) 7

3.2.1Contact 7

3.2.2The Roles of the EURL 8

3.3The UK National Reference Laboratory 9

3.3.1Contact 9

3.3.2The roles of the Anisakis NRL 9

5.Industry and Consumer 10

5.1Industry Requirements 10

4.1.1 Regulation (EC) No 853/2004 [extract]. 10

5.2Analysis Requirements within Industry 11

4.2.1 Time of Sampling 11

4.2.2 Sampling Method 11

4.2.3 Preparation of Samples 11

4.2.4 Additional Advice 12

5.3Information for Consumers 12

5.4Anisakiasis: Further Information, 12

6.Official Control Laboratories 14

6.1OCL Requirements 14

6.2Method for Detection of Anisakidae larvae in fish fillets using ultraviolet transillumination 14

5.2.1 Scope 14

5.2.2 Equipment 14

5.2.3 Method 15

Results 16

6.3Summary of detection methods for Anisakidae larvae in fish fillets 17

5.3.1 Scope 17

Receipt of samples 17

5.3.2 Digestion 17

5.3.3 Compression System 18

5.3.4 Candling 18

6.4Further Advice for OCLs 19

7.References 20

8.Annex 22

List of European (parasite) NRLs (ISS, 2015) 22

3.Introduction

Anisakiasis is the term used to describe infections of humans with larval stages of ascaridoid nematodes within the family Anisakidae and occasionally within the family Raphidascarididae. The life cycle of ascaridoid nematodes involves the production of eggs by adult females in mammalian hosts which are shed into the water. Following development in the egg, including at least one moult, second stage larvae hatch into the water column which are eaten by an invertebrate hosts (normally a euphausid crustacean). Transmission to other hosts, such as fish and cephalopods is possible via ingestion of infected euphausids.

Infections in humans may occur when raw, undercooked or ill-prepared fishery products are consumed in which viable nematode larvae occur. Whilst the vast majority of infections reported in humans have been associated with Anisakis simplex and Pseudoterranova decipiens, there have been a number of instances were other species of nematodes have been implicated.
There is an obligation on the part of The Food Standards Agency as the designated Competent Authority for the purposes of ‘Regulation (EC) 882/2004’ on Official Feed and Food Controls in the UK to ensure that any risk to public health is reduced, prevented or eliminated. This obligation includes the designation of European Reference Laboratories for the analysis or examination of feed and food, including Anisakis spp. in fish products.

4.Administration

1. Introduction

There is a requirement under Regulation (EC) No 882/2004 to establish a network of laboratories within the European Union for the purpose of the monitoring, control, and eradication of animal diseases which are known to have human health impacts. Where applicable, controls should be carried out using appropriate techniques developed for that purpose, including routine industry practice and sampling and the testing of samples¹.

The network of laboratories designated by the European Commission will include a European Union Reference Laboratory and National Reference Laboratories. Such laboratories will work in collaboration with testing laboratories to ensure validity of testing procedures and appropriate training.

2. The European Union Reference Laboratory (EURL)

1. Contact

European Reference Laboratory for Parasite,

Gastroenteric and Tissue Parasitic Diseases Unit,

Department of Infectious, Parasitic and Immunomediated Disease,

Instituto Superiore di Sanità,

Viale Regina Elena 299,

00161 – Roma (Italy),

EURLP Director: Edoardo Pozio

Phone: [phone],

Email: [e-mail]

2. The Roles of the EURL²

Regulation (EC) No 882/2004 defines tasks, duties and requirements for EURLs. The Commission can establish new EURLs or change designation of existing ones.
EU Reference Laboratories (EURLs) must aim to ensure high-quality, uniform testing in the EU and support Commission activities on risk management and risk assessment in the area of laboratory analysis.
Provide National Reference Laboratories (NRLs) with analytical methods and diagnostic technics, and coordinate their application;
Train staff from National Reference Laboratories;
Provide the Commission with scientific and technical expertise in relation to laboratory analysis (e.g. assist actively in the diagnosis of animal disease outbreaks);
Collaborate with the competent laboratories in non-EU countries.

3. The UK National Reference Laboratory

1. Contact

UK National Reference Laboratory for Anisakis (NRL) ³,

Centre for Environment, Fisheries, and Aquaculture Science,

Barrack Road,

The Nothe,

Weymouth,

Dorset,

DT4 8UB

United Kingdom.

NRL Director: Dr Stephen W. Feist.

Phone: [phone]

Email: [e-mail]

2. The roles of the Anisakis NRL

Provided scientific and technical assistance to the Official [Feed and Food] Control Laboratories and the Competent Authority, as appropriate;
Co-ordinate the activities of Official Control Laboratories (OCLs) responsible for the analysis of samples in accordance with Article 11 of Regulation (EC) No 882/2004;
Organise comparative tests between the OCLs and ensure appropriate follow-up of such comparative testing;
Disseminate information and documentation, including the maintenance of Standard Operating Procedures and guidance documents;
Provide up to date information to the FSA;
Represent the UK at EURL meetings and working groups, as appropriate

5.Industry and Consumer

5.1Industry Requirements

Regulation (EC) No 882/2004 states that “… official controls are carried out regularly, on a risk basis…”. This statement is particularly relevant for Anisakis as there is currently no requirement or program of work for the routine monitoring or reporting of Anisakidae larvae within commercial fish production or harvest. The industry has a good record of using best practice alongside their own internal quality control procedures in order to ensure that parasites do not enter the food chain and they continue to maintain compliance with relevant EU legislation (Regulation (EC) No 853/2004⁴) regarding the hygienic production of foodstuffs. This legislation contains a specific reference to the processing of fish which may be hosts to parasites, including Anisakis spp (see below).

4.1.1 Regulation (EC) No 853/2004 [extract].

‘Laying Down Specific Hygiene Rules for on the Hygiene of Foodstuffs’

Chapter iii: Requirements for Establishments, Including Vessels, Handling Fishery Products.

Section D: Requirements Concerning Parasites.
1. The following fishery products must be frozen at a temperature of not more than -20 °C in all parts of the product for not less than 24 hours; this treatment must be applied to the raw product or the finished product:

(a) fishery products to be consumed raw or almost raw;

(b) fishery products from the following species, if they are to undergo a cold smoking process in which the internal temperature of the fishery product is not more than 60 °C:

(i) herring;

(ii) mackerel;

(iii) sprat;

(iv) (wild) Atlantic and Pacific salmon; and

(c) marinated and/or salted fishery products, if the processing is insufficient to destroy nematode larvae.
2. Food business operators need not carry out the treatment required under paragraph 1 if:

(a) epidemiological data are available indicating that the fishing grounds of origin do not present a health hazard with regard to the presence of parasites; and

(b) the competent authority so authorises.
3. A document from the manufacturer, stating the type of process they have undergone, must accompany fishery products referred to in paragraph 1 when placed on the market, except when supplied to the final consumer.

5.2Analysis Requirements within Industry

Should there either a legislative or industry lead drive for formal monitoring and identification of parasites, samples of individual larvae/worms and/or whole fish can be sent to any of the laboratories which have been deemed proficient by the guidelines laid out by competent authority.

4.2.1 Time of Sampling

Sampling for the purpose of continual monitoring should be undertaken, where practical, on as random a basis as possible with respect to environmental conditions. If a sample is reactionary then it should be dealt with as quickly as possible.

4.2.2 Sampling Method

Fish sampling should be undertaken using the method normally used for the commercial sampling of the host species in question, as this can influence potential contamination. If sampling individual worms, care should be taken to not damage the delicate structures of the organism, as this may make gross taxonomic identification problematic.

4.2.3 Preparation of Samples

Whole fish or fillet samples should be as clean as possible with the skin left on. Samples should be individually bagged, labelled as appropriate, and placed into a temperature stable box for transport. Providing a courier can be arranged to deliver the sample(s) to a laboratory within 24-48 hours, a normal cool box containing freezer packs or bags of ice is sufficient. Individual worms should be stored in 100% ethanol or other high concentration alcohol. The specific requirements/procedures of each testing laboratory may differ, so it is suggested that such details are confirmed prior to sampling and shipping.

4.2.4 Additional Advice

Additional advice on sampling can be obtained from either the EURL or the CRL. Please see contact details in section 2 or the Annex of this document.

5.3Information for Consumers⁵

There is considerable legislation setting out the legal requirement for the industrial production of fishery products for human consumption. However, there is no requirement for private or domestic consumers to comply with this legislation i.e. wild salmon caught from UK rivers. As such the FSA advise consumers to ensure that such products are safe to eat. Consumers should:

Visually inspect the fish or fillet and remove parasites. Those fish which are found to be obviously or heavily contaminated should not be consumed.
If wild salmon is to be eaten raw or almost raw it should be frozen for at least 24 hours, at a temperature of at least -20°C. This will ensure that any non-visible parasites or undetectable larvae are destroyed. This advice also applies to produce which will undergo cold smoking or salting.
If freezing as described above is not possible, it is highly advisable to cook the flesh thoroughly. 70°C for two minutes will render any contamination safe.
As there is no infallible method of detecting and removing larvae, this advice is particularly relevant for pregnant women, elderly people, and the immunocompromised where ingestion of live parasites from fish could pose a serious health risk.

5.4Anisakiasis: Further Information^{6, 7}

Clinical infections of Anisakidae larvae within Europe are proportionally rare. In most cases [live] ingested larvae die and are digested within the gastrointestinal tract. However, in some cases the parasite migrates into the intestinal wall where it elicits severe inflammatory reactions, and clinical symptoms. The location of the infection may be gastric, intestinal or ectopic, and presents as: peptic ulcer disease; acute abdomen; a bowel obstruction; or as abdominal pain, either minor or intense, with or without vomiting. Due to the diversity of possible symptoms, the disease is often misdiagnosed. Humans are actually an accidental host to such parasites, and, as such, the organism cannot continue its lifecycle following ingestion by a human. There is no risk of person-to-person infection.

The occurrence of clinical Anisakiasis is generally prevented within European Countries as a result of the control measures currently in place. However, ingestion of Anisakis can also cause anaphylaxis and/or urticaria in certain patients, regardless of the parasite establishing, or even surviving, within the gastrointestinal tract. The current control measures, other than direct removal, do not reduce the risk of allergic reaction, as the specific allergens are thermostable. The epidemiology of allergic reactions to Anisakis is not well established, and is most likely underreported.
Consumer, industry, and medical awareness is key to the prevention of serious allergic reaction to this parasite.

6.Official Control Laboratories

4. 
   
   

6.1OCL Requirements

Participation in relevant proficiency testing is an essential part of a laboratory's quality assurance system. OCLs undertaking the examination of samples of fish for the presence of Anisakis nematodes are encouraged to take part in an appropriate external quality assessment (EQA) scheme as well as any proficiency testing that their respective NRL may provide.
The Anisakis EQA scheme is based on the use of artificial samples. The NRL distributes proficiency-test material in the form of either fish muscle containing anisakid nematodes. Distribution is undertaken annually if required. Laboratories that are part of the NRL network will be contacted prior to each distribution and will be expected to participate.
At the time of writing this document the screening of fish produce for Anisakis is not a legislative requirement. As such the OCLs within the network are not specifically required to undertake proficiency testing.

6.2Method for Detection of Anisakidae larvae in fish fillets using ultraviolet transillumination⁸

5.2.1 Scope

This SOP describes a non-GLP technique for the screening of fish fillets for Anisakidae larvae using a compression and UV transillumination method. After isolation, parasites could either be submitted for molecular or traditional morphological analysis.

5.2.2 Equipment

Fillet Knife;

Strong Clear Plastic Bag(s);

Compression System;

UV Transilluminator (Fisher Scientific ref: 12843938);

Bench top UV Darkroom w/ UV down light (C-65 modular darkroom, Fisher Scientific ref: 11788201) (Figure 1).


Figure - Bench top UV Darkroom mounted on UV transilluminator
PPE and Clinical Waste Bag;

Disinfectant i.e. Virkon® or Ethanol;

Freezer, -20°C if possible;

Additional Sample Containers (if required).

5.2.3 Method

Receipt of samples

Samples may be received as whole fish or as pre-prepared fillets or tissue sections. Samples should be immediately assessed for their condition and assigned a specific reference number using a suitable Database. If immediate screening is not possible, samples can be stored in a fridge (approx. 4°C) for a maximum of 48 hours.

Sample Preparation

If fish samples have been received whole carefully fillet using a suitable knife ensuring that muscle tissue is removed from the carcass as cleanly as possible. Fillet both sides of the fish. Place each fillet into a clear plastic bag ensuring there is sufficient space in the bag for compression. Using a suitable system, squeeze both fillets until they are approximately 1-2mm thick. Freeze the fillets (at -20°C if possible). The skin can be removed prior to compression or after freezing as required

Sample Screening

Place the individual compressed and frozen fillets into a bench top UV transilluminator and examine using UV light (λ 366nm). CAUTION – UV light is damaging to eyes, please ensure suitable precautions/PPE. Anisakidae larvae will fluoresce as white spots or individual worms (figure 2). Larvae can then be removed for other diagnostic methods if required.

Figure - Anisakis encapsulated in fish fillet, under illumination with an ultraviolet light source. Individual parasites appear as bright/fluorescent inclusions within the flesh (image from Wootten and Bron, 2008)

Results

Results should be officially recorded immediately after screening is complete. Images can be taken using a standard digital camera held against the eyepiece of the Bench top darkroom. Results should be reported to the customer/stakeholder within 48 hours.
Disposal and Decontamination

Waste tissue should be placed into a clinical waste bag and disposed of using an authorized route. All equipment should be sterilised using a suitable disinfectant.

6.3Summary of detection methods for Anisakidae larvae in fish fillets⁹

5.3.1 Scope

This SOP provides a brief summary of other methods for the detection of Anisakis larvae in fish muscle tissue as detailed by the ‘European Union Reference Laboratory for Parasites (EURL)’ for use during proficiency testing (PT) and routine screening. In some instances these methods have been modified using those described by Karl and Leinemann (1993). A study by the same paper (Karl and Leinemann, 1993) showed that compression and examination under ultraviolet (λ 366nm) was at least as effective as the digestion method described below. For a detailed description of this method as used by Cefas, please see section 5.2.1. After isolation where possible, parasites could either be submitted for molecular or traditional morphological analysis.

Receipt of samples

Samples may be received as whole fish or as pre-prepared fillets or tissue sections. Samples should be immediately assessed for their condition and assigned a specific reference number using a suitable Database. If immediate screening is not possible, samples can be stored in a fridge (approx. 4°C) for a maximum of 48 hours.

5.3.2 Digestion

Equipment

Hydrochloric acid;

Distilled water;

Pepsin;

3L beaker;

Blender;

Knife or Scissors;

Heated magnetic stirring plate;

Sieve.


Method

If fish samples have been received whole carefully fillet using a suitable knife ensuring that muscle tissue is removed from the carcass as cleanly as possible. Fillet both sides of the fish. Add 16ml ±0.5ml of hydrochloric acid (HCl) to 2.0L of pre-heated (46-48°C) distilled water in a 3L beaker. Maintain heating and stir using a magnetic stirrer. Add 10 ±0.2g of pepsin to the HCl solution. Blitz the fillet sample in a blender for 1-2 seconds – ensure that liquidation does not occur. Immerse the flesh in the HCl/pepsin solution. Ensure that all flesh is removed from the blender by rinsing with a small amount of HCl/pepsin solution. Cover the beaker with foil, reduce temperature to 44-46°C and ensure continued stirring does not result in splashing. Once the muscle tissue has completely dissolved the solution should be poured through a sieve – the filtrate must be disposed of through an approved route. Anisakidae should be retrieved from the sieve for further processing as required.

5.3.3 Compression System

Equipment

Fillet knife;

Compressorium of suitable analogue;

Stereoscope or low powered microscope (5-10X).

Method

If fish samples have been received whole carefully fillet using a suitable knife ensuring that muscle tissue is removed from the carcass as cleanly as possible. Fillet both sides of the fish. If using a traditional compressorium the fillets will need to be sliced thinly prior to mounting within the compressorium. Fillets can be examined whole by placing between two strong sheets of plexiglass (strengthened glass can be used but is not recommended) ensuring there is sufficient space in the compression. Using a suitable manual or hydraulic system, squeeze both fillets until they are approximately 1-2mm thick. Secure using clamps. Using a microscope with an up-light capability scan each fillet or compressorium prep for larvae.

5.3.4 Candling

Equipment

Fillet knife;

Light box.
Method

If fish samples have been received whole carefully fillet using a suitable knife ensuring that muscle tissue is removed from the carcass as cleanly as possible. Fillet both sides of the fish. Fillets should be placed between two strong sheets of plexiglass (strengthened glass can be used but is not recommended) ensuring there is sufficient space for compression. Using a suitable manual or hydraulic system, squeeze both fillets until they are approximately 3-4mm thick. Secure using clamps.

Alternatively, fillets could be examined without compression (however low penetration of visible light makes this problematic); or sliced thinly. Place each fillet (or section) on the light box using approx. 1500 lux.. Worms will appear as dark shadows in the flesh, these can be removed for subsequent analysis as required.
Results

Results should be officially recorded immediately after screening is complete. Images can be taken using a standard digital camera held against the eyepiece of the Bench top darkroom. Results should be reported to the customer/stakeholder within 48 hours.

Disposal and Decontamination

Waste tissue should be placed into a clinical waste bag and disposed of using an authorized route. All equipment should be sterilised using a suitable disinfectant.

6.4Further Advice for OCLs

Further information is available for OCLs from both the EURLP¹⁰ and the UK NRL¹¹. Please see contact details in section 2 or the Annex of this document.

7.References

Crown Copyright (2007). Wild salmon and the Anisakis parasite: Guidance. Accessed online 4/3/2015 from: http://tna.europarchive.org/20130129064346/http://www.food.gov.uk/business-industry/guidancenotes/hygguid/guidsalmonanisakis

European Commission. (2015a). Food Safety, Legislation, References Laboratories. Accessed online 4/3/2015: http://ec.europa.eu/food/safety/official_controls/legislation/ref-labs/index_en.htm
European Union Reference Laboratory for Parasites (2014), ‘Form 3: PT on DETECTION OF ANISAKIDAE L3 LARVAE IN FISH FILLETS – Procedure’. Department of Infectious, Parasitic and Immunomediated Diseases, Unit of Gastroenteric and Tissue Parasitic Diseases . Istituto Superiore di Sanità
Hill, T. (2014b) Method for Detection of Anisakidae larvae in fish fillets using ultraviolet transillumination. United Kingdom Anisakis NRL SOP NRL002. Centre for the Environment, Fisheries and Aquaculture Science.
Hill, T. (2014b) Summary of detection methods for Anisakidae larvae in fish fillets. United Kingdom Anisakis NRL SOP NRL001. Centre for the Environment, Fisheries and Aquaculture Science.
Instituto Superiore di Sanità (ISS). (2015). List of NRLs: EURLP website. Accessed 4/3/2015 from: http://www.iss.it/crlp/index.php?lang=2&id=51&tipo=11
Karl, H.; Leinemann, M. (1993). A fast and quantitative detection method for nematodes in fish fillets and fishery products. Arch. Lebensmittelhyg. 44(5): 124-125 In: Archiv für Lebensmittelhygiene.: Hannover. ISSN 0003-925X
Regulation (EC) No 882/2004 of the European Parliament and of the Council of April 2004.
Wootten, R. and Bron, J. (2008) 'Worms in Fish’ Aquaculture News 34. Published by the INSTITUTE OF AQUACULTURE, University of Stirling, Stirling, FK9 4LA. Scotland UK. ISSN 1357-1117
Audicana, M.T. & Kennedy, M.W., 2008. Anisakis simplex: from obscure infectious worm to inducer of immune hypersensitivity. Clinical microbiology reviews, 21(2), pp.360–79, table of contents. Available at: http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=2292572&tool=pmcentrez&rendertype=abstract [Accessed February 15, 2015].
European Union (EU). (1998). Opinion of the Scientific Committee on Veterinary Measures relating to Public Health - Allergic reactions to ingested Anisakis Simplex antigens and evaluation of the possible risk to human health - 27 April 1998 http://ec.europa.eu/food/fs/sc/scv/out05_en.html

8.Annex

List of European (parasite) NRLs (ISS, 2015)

AUSTRIA
The Austrian Agency for Health and Food Safety, Institute of Veterinary Medicine - Technikerstraße 70, A-6020 Innsbruck
Contacts: Karl Schöpf; Walter Glawischnig; tel. [phone]; fax [phone].
Web site AGES

BELGIUM AND LUXEMBOURG

Prince Leopold Institute of Tropical Medicine (ITG) - Nationalestraat 155, B 2000 Antwerpen
Contacts: Pierre Dorny, Marleen Claes (Trichinellosis, Cysticercosis, Echnococcosis); tel. [phone]; fax: [phone].
Web site: ITG

BULGARIA
National Diagnostic and Research Veterinary Institute, Parasitic Zoonoses Laboratory - 15A Pencho Slaveikov blvd, 1606 Sofia

Contacts: Nikolay Lalkovski Tsvetanov; Tanya Kostova; tel. [phone]; fax [phone].

CYPRUS
State Veterinary Laboratory, Veterinary Services - 1417 Athalassa Nicosia

Contacts: Costantinos Economides, Andreas Phylactou ; tel. [phone]; fax [phone].

CROATIA
Faculty of Veterinary Medicine University of Zagreb - Heinzelova 55 Zagreb 10000 Zagreb

Contact: Albert Marinculic; tel. [phone]; fax [phone].
Croatian Veterinary Institute, Veterinary Department Vinkovci - J. Kozarca 24, 32100 Vinkovci
Contacts: Davor Balić; tel. [phone]; fax [phone].

CZECH REPUBLIC

State Veterinary Institute, Department of Pathological Anatomy - Jakoubka ze Stribra 1, 779 00 Olomouc
Contact: Jiri Harna; tel. [phone]; Mobile:[phone]; fax. [phone].
Web site: SVUO

DENMARK
National Veterinary Institute, Technical University of Denmark, Section for Immunology and Parasitology" Bülowsvej 27, DK-1790 Copenaghen V

Contact: Heidi L. Enemark; tel. [phone].
Web site: DFVF

ESTONIA

Estonian Veterinary and Food Laboratory - Kreutzwaldi 30, 51006 Tartu
Contact: Age Kärssin; tel. [phone]; fax [phone].
Web site: VAFL

FINLAND
Finnish Food Safety Authority Evira, Oulu Research Unit - PO. Box 517 (Samatie 15) 90101 Oulu

Contact: Varpu Hirvela-Koski; tel. [phone]; fax: [phone].
Web site: EVIRA

FRANCE
French agency for food, environmental and occupational health safety ANSES - Animal Health Laboratory" 23 avenue du Général de Gaulle 94706 Maisons-Alfort cedex

Contact: Isabelle Vallée (Trichinellosis and Anisakiasis); tel. [phone]; fax: [phone].
ANSES, Nancy laboratory for rabies and wildlife - Technopôle Agricole et Vétérinaire - BP 40009, 54220 Malzéville
Contact:Franck Boue (Echinococcosis); tel. [phone]; fax: [phone].
Web site: ANSES

GERMANY
Federal Institute for Risk Assessment - Diedersdorfer Weg 1, D - 12277, Berlin

Contact: Karsten Noeckler (Trichinellosis); tel. [phone]; fax: [phone].
Web site: BfR.
Friedrich-Loeffler Institute, Federal Research Institute for Animal Health - Boddenblick 5a, 17493 Greifswald, Insel Riems
Contact: Franz J. Conraths (Echinococcosis); tel. [phone]; fax: + 49 383 51 72/19, [phone].
Web site: FLI
Max Rubner-Institut, Federal Research Institute of Nutrition and Food - Haid und Neu Str. 9, D-76131 Karlsruhe
Contact: Karl Horst (Anisakiasis ); tel. [phone]; fax: [phone].
Web site: BfEL

GREECE
Center of Athens Veterinary Institutions, Department of Parasitology" 25, Neapoleos St, 15310 Ag. Paraskevi, Athens

Contact: Sofia Boutsini (Toxoplasmosis and Trichinellosis); tel. [phone]; fax: [phone].

HUNGARY
Central Veterinary Institute" Tábornok u. 2, H-1149, Budapest

Contact: Tamas Sréter; tel. [phone]; fax: [phone].
Web site: OAI

ICELAND
Institute for Experimental Pathology, University of Iceland - v/Keldnaveg, IS-112 Reykjavík

Contact: Vala Friðriksdóttir; tel. [phone] direct 5855150; fax [phone].
Web site: The Institute For Experimental Pathology, University of Iceland, KELDUR

IRELAND
Veterinary Laboratory, Department of Agriculture & Food Laboratories - Young's Cross, Celbridge, County Kildare

Contact: John Egan; tel. [phone]; Fax: [phone].
Central Meat Control Laboratory, Veterinary Laboratory, Department of Agriculture & Food Laboratories - Backweston, Celbridge, Co. Kildare
Contact: William Byrne.
Web site: Department of Agriculture & Food

ITALY
Section of Gastroenteric and Tissue Parasitic Diseases, Istituto Superiore di Sanità - Viale Regina Elena 299, 00161 Rome

Contact: Edoardo Pozio (Trichinellosis); tel. [phone]; fax + 39 0649903561 .
Web site: ISS
Sicily Experimental Zootechnic Institute
Via Gino Marinuzzi 3, 90129 Palermo - Contact: Vincenzo Ferrantelli, Angela Alongi (Anisakiasis); tel. [phone]; fax [phone].
Via Passo Gravina, 195 - 95125 Catania - Contact: Maria Fausta Marino (Toxoplasmosis); tel. [phone]; fax [phone].
Web site: IZSSI
Sardinia Experimental Zootechnic Institute - Via Duca degli Abruzzi 8, 07100 Sassari
Contact: Giovanna Masala (Echinococcosis); tel. [phone]; fax [phone].
Web site: IZS-SARDEGNA

LATVIA

Laboratory of Food and Environmental Investigations (LFEI), Parasitology Division, National Diagnostic Centre - Lejupes street 3, LV-1076 Riga.
Contact: Muza Kirjusina (Trichinellosis, Echinococcosis, Anisakiasis); tel. [phone]; fax [phone].
Web site:PVD NDC

LITHUANIA

National Food and Veterinary Risk Assessment Institute - J. Kairiukscio 10, LT- 08409, Vilnius
Contact: Ieva Grazenaite; tel [phone]; fax: [phone].
Web site: NMVRVI

MALTA
Food Health and Diagnostics Laboratory, Veterinary Regulation, Fisheries Conservation and Control Division - Civil Abattoir Square, Albert Town, Marsa

Contact:Susan Chircop; tel. [phone].
Veterinary Laboratory, Department of Food Health & Diagnostics, Veterinary Affairs and Fisheries Division - Albertown, Marsa
Contact: Albert Gambin (Trichinellosis); tel. [phone]; fax: [phone].
Web site: Food and Veterinary Regulation Division
Centre for Infections Health Protection Agency - 61, Colindale Avenue, London NW9 5HT, UK
Contact: E. John Threlfall (Echinococcosis, Anisakiasis); tel. [phone]; fax: [phone].
Web site: HPA

NETHERLANDS

Microbiological Laboratory for Health Protection, National Institute of Public Health and the Environment (RIVM) Antonie van Leeuwenhoeklaan (P.O. Box 1) 9, 3720 Ba, Bilthoven
Contacts: Joke W. B. van der Giessen; Frits Franssen (Trichinellosis); tel. [phone]; fax: [phone].
Web site: RIVM

POLAND
National Veterinary Research Institute" Partyzantow 57, 24-100, Pulawy

Contacts: Department of Parasitology: Irena Ziomko ; Jacek Osek; Jacek Karamon (echinococcosis); Jacek Sroka (toxoplasmosis); tel. [phone]; fax: [phone].
Department of Food Hygiene: Miroslaw Rozycki (trichinellosis).
Web site: PIWET

PORTUGAL
National Laboratory of Veterinary Research - Estrada de Benfica 701, 1649-016, Lisboa

Contact: Jacinto Gomes .
Web site: LNIV
Instituto Nacional de Saúde Dr. Ricardo Jorge, Department of Infectious Diseases - Av. Padre Cruz, 1649 - 016 Lisboa
Contact: Maria João Gargate; tel. [phone]; fax: [phone].
Web site: INSA

ROMANIA
Institute for Diagnosis and Animal Health - Staicovici str. 63, sect. 5, RO- 76202 Bucarest.

Contacts: Nicolae Stefan;tel. [phone].
Department of Aquatic Animals and Useful Insects Health: Mihaela Costea; tel. [phone]; fax [phone].
Institute of Hygiene and Public Veterinary Health - Str. Campul Mosilor no.5, district 2, 021201 Bucharest.
Contacts: Niculai Poparlan, Andrei Nicolau (Trichinellosis); tel. [phone]; fax [phone].
Web site: IISPV

SLOVAK REPUBLIC

State Veterinary and Food Institute Bratislava - 15, Botanická 842 52 Bratislava
Contact: Daniela Valentova (Trichinellosis, Echinococcosis and Anisakiasis); tel. [phone].
Web site: SVUBA

SLOVENIA
University of Ljubljana, Veterinary Faculty, Laboratory of Parasitology" 60, Gerbičeva, 1000 Ljubljana

Contact: Janez Posedi (Trichinellosis, Echinococcosis and Anisakiasis); tel. [phone]; fax: [phone].
Web site: VF

SPAIN
Central Laboratory of Animal Health - Camino del Jau s/n, 18320 Santa Fe, Granada

Contacts: Fulgencio Garrido Abellán, Agustina Perales Flores; tel. [phone]; fax: [phone].
Food National Centre. Spanish Agency for Consumer Affairs, Food Safety and Nutrition (AECOSAN) - Carretera a Pozuelo Km 5.1. 28220 Majadahonda.
Contact: Carmen Blanco Vidal (Trichinellosis, Echinococcosis and Anisakiasis); tel. [phone]; fax [phone].
Web site: AECOSAN

SWEDEN
National Veterinary Institute, SVA, Dept. of Virology, Immunobiology and Parasitology, Section for Parasitology - Ullsv 2, 751 89 Uppsala

Contacts:Eva Osterman, Anna Lundén; tel. [phone]; fax [phone].
Web site: SVA

UNITED KINGDOM

Regional Laboratory, Animal Health and Veterinary Laboratories Agency(AHVLA) - Rougham Hill, Bury St Edmunds, Suffolk, IP44 2RX.
Contacts: Paul Todd, Paul Harris; tel. [phone]; Fax: [phone].
Web site: AHVLA
OIE Collaborating Centre: Information on Aquatic Animal Diseases, Cefas Weymouth Laboratory - Barrack Road, The Nothe, Weymouth, Dorset, DT4 8UB.
Contacts: Stephen W. Feist; tel. + 44 (0)1305 206662; Fax: + 44 (0)1305 206601.
Web site: CEFAS.

1^ Regulation (EC) No 882/2004

2^ European Commission (2015a).

3^ http://www.cefas.defra.gov.uk/anisakis.aspx

4^ Regulation (EC) 953/2004

5^ This section is adapted from an article released by the Fish and Shellfish Hygiene Branch of the FSA. Crown Copyright (2007). http://tna.europarchive.org/20130129064346/http://www.food.gov.uk/business-industry/guidancenotes/hygguid/guidsalmonanisakis

6^ Audicana, M.T. & Kennedy, M.W., 2008. Anisakis simplex: from obscure infectious worm to inducer of immune hypersensitivity. Clinical microbiology reviews, 21(2), pp.360–79, table of contents. Available at: http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=2292572&tool=pmcentrez&rendertype=abstract [Accessed February 15, 2015].

7^ Parts of this section have been adapted from an article released by the European Commission. (EU, 1998). http://ec.europa.eu/food/fs/sc/scv/out05_en.html

8^ Ref: NRL001 (Hill, T. 2014a)

9^ Ref: NRL001 (Hill, T. 2014b)

10^ http://www.iss.it/

11^ http://www.cefas.defra.gov.uk/anisakis.aspx

UK Anisakis Reference Laboratory: Code of Practice/Guidance Document – DOC32 Revision 1

Printed versions of this document are uncontrolled unless signed by the Quality Department