National Reference Laboratory

Disease Transmission
Fragments of animal origin in animal feed indicate that animal material has been used. This significantly increases the risk of transmitting existing diseases such as Bovine Spongiform Encephalopathy (BSE or mad cow disease) via prions.

BSE spreads primarily through the recycling of animal remains in animal feed. Feed bans (Regulation (EC) No. 999/2001: the feed ban) minimize the risk of contamination, and monitoring programs (Regulation (EC) No. 152/2009) enforce this ban.

Animal Proteins
WFSR is the only official animal protein control laboratory in the Netherlands. We monitor animal feed for the presence of fragments of animal origin for the Netherlands Food and Consumer Product Safety Authority (NVWA).

Official Methods
Visual techniques, primarily microscopic examination, and DNA-based detection are the primary methods for detecting the presence of animal material. These methods can be supported by protein detection.

WFSR is part of the EU Network for Animal Proteins (EURL-AP) and also participates in the proficiency tests organized by the EURL-AP.

GMOs are plants, animals, or microorganisms (and derivatives) whose DNA has been modified through biotechnology. A crop may, for example, be made resistant to a herbicide or insect pest. The added DNA comes from other organisms or is self-generated.

The GMO Register lists EU-approved GMOs. The Netherlands enforces EU regulations for GMOs based on, among other things:

• Regulation (EC) No. 1829/2003 on food and feed

• Regulation (EC) No. 1830/2003 Traceability and Labeling

The developer also provides an event-specific detection method and reference material to the EU Reference Laboratory for GM Food and Feed (EURL-GMFF) for enforcement purposes. The European Network of GMO Laboratories (ENGL), of which WFSR is a member, evaluates these methods. WFSR also develops its own detection methods.

New Genomic Techniques (NGTs) are also used to create GMOs, for example, targeted mutagenesis using CRISPR/Cas. This modifies or removes individual DNA building blocks. WFSR monitors developments regarding the detectability of these methods.

Marine biotoxins are produced by algae and can accumulate in edible shellfish that filter the algae from the water, such as mussels and oysters. Consumption of contaminated shellfish can lead to serious poisoning symptoms such as memory loss, vomiting, and paralysis. To ensure food safety, European and Dutch guidelines have been established, and toxin concentrations in shellfish from official production areas are regularly determined. In the Netherlands, legislation applies to the following toxins: domoic acid, saxitoxins, okadaic acid and dinophysistoxins, azaspiracids, yessotoxins, and tetrodotoxin (TTX).

Dutch legislation applies to TTX (which causes, among other things, paralysis symptoms) (Commodities Act Policy Rule on TTX in live bivalve molluscs 2022); the other toxins are regulated at the European level (Regulation (EC) No. 853/2004 - Annex III, Section VII, Chapter V).

Chemical methods and cell testing
WFSR is an expert in analyzing marine biotoxins in shellfish. We have analytical methods for regulated toxins (Regulation (EU) 2019/627, Regulation (EC) No. 2021/1709). We can also determine a wide range of non-regulated toxins using various chemical methods and cell tests.

The NRL Milk and Dairy Products ensures the quality of official analyses of dairy products in the Netherlands by examining:

Total aerobic plate count
Most bacteria in raw milk do not pose a health risk to consumers, but a high bacteria count is a sign that hygiene regulations are not being followed for that batch of milk.

Somatic cell count
Somatic cells (mammal cells) can enter the milk when cows suffer from mastitis. A high cell count is not good for the quality of the milk, but also indicates animal welfare.

Milk Enzyme Alkaline Phosphatase
A specific heat treatment, which complies with the requirements for pasteurization, ensures that this milk enzyme is completely inactivated. This allows us to determine whether a producer has pasteurized sufficiently. It also allows us to determine, for example, whether farmhouse cheese is indeed made from unpasteurized milk, as required for such cheese.

Regulation (EC) No 853/2004 - Annex III, Section IX sets maximum levels for these three areas.

NRLs ensure that these tests are carried out in an equivalent manner in all EU countries.

Expertise
WFSR has specific expertise in the analysis of heavy metals and toxic elements, as well as nitrogen compounds, such as melamine, cyanuric acid, nitrate, nitrite, and N-nitrosamines, in food and feed. WFSR also has knowledge of and experience with metals and nitrogen compounds-related legislation, method development and validation, quality control, and the assurance of government-appointed laboratories.

Independent quality assurance
As part of independent quality assurance and the validation and comparison of test methods, WFSR participates in collaborative studies organized by the EURL (European Reference Laboratory) and third parties, as well as in interlaboratory studies. The EURL for metals and nitrogen compounds is the Technical University of Denmark (DTU) in Copenhagen.

Reference Substances and Reagents
For questions about reference substances and reagents that can be used for analyses within the scope of the NRL for metals and nitrogen compounds, please contact the NRL via the contact form on this website.

WFSR conducts official inspections to ensure compliance with food and feed legislation, animal health and welfare, plant health, and plant protection products.

Natural Toxins
Mycotoxins (produced by molds) and plant toxins occur naturally in food and feed and can harm human and animal health. Although their presence cannot be completely prevented, EU legislation sets maximum levels to protect consumers and animals. The monitoring covers both regulated and emerging mycotoxins, including aflatoxins (AFL), ochratoxin A (OTA), deoxynivalenol (DON), fumonisins (FB), zearalenone (ZON), patulin (PAT), T-2/HT-2 toxins, citrinin (CIT), alternaria toxins, and ergot alkaloids (EA). Plant toxins include pyrrolizidine alkaloids (PA), tropane alkaloids (TA), glycoalkaloids (GA), quinolizidine alkaloids (QA), opium alkaloids (OA), cannabinoids (THC), and cyanogenic glycosides (CNG).

Analysis and Support
WFSR specializes in analyzing these toxins, developing and validating methods, providing quality assurance for official laboratories, identifying emerging risks, and providing scientific and technical support to competent authorities.

Persistent organic contaminants (POPs) are poorly biodegradable and therefore persist in the environment for extended periods. They can accumulate in food and pose risks to humans due to their toxicity. Reliable research is therefore essential. WFSR experts play a leading role in the PFAS and CP Core Working Groups of the European Union Reference Laboratory for POPs. Within these networks, EU laboratories exchange knowledge to harmonize analytical methods and establish performance criteria in European legislation.

WFSR investigates, among other things:

• Dioxins and polychlorinated biphenyls (PCDD/Fs and PCBs)

• Per- and polyfluoroalkyl substances (PFAS, such as PFOS and PFOA)

• Chlorinated paraffins (CPs/PCAs)

• Brominated flame retardants (BFRs/BCons)

• Other halogenated POPs

The list of POPs in use is constantly evolving. Because toxicity and environmental effects are often unknown, WFSR is developing new analytical methods together with European partners to provide the government with reliable information in a timely manner.

WFSR performs this task within the framework of official monitoring and enforcement in accordance with Regulation (EU) No. 2023/915. The scope of this NRL task includes the following process contaminants:

• Acrylamide

• Furans (including furan and 2- and 3-methylfuran)

• 2- and 3-MCPD esters and glycidyl esters, as well as free 2- and 3-MCPD

• Polycyclic aromatic hydrocarbons

• Mineral oils

WFSR, as the NRL, is responsible for To maintain its expertise in the above-mentioned areas in accordance with Regulation (EC) No. 333/2007. WFSR maintains regular contact with the European Reference Laboratory (EURL) for this purpose, including by attending workshops and participating in collaborative studies. This ensures the required quality and reliability of the NRL's analytical methods. The EURL for process contaminants is the Technical University of Denmark (DTU). WFSR also has an advisory role to the government.

In the Netherlands and Europe, the use of growth hormones in livestock is prohibited. Furthermore, food products may not contain residues of unregistered animal treatment products. For approved products such as antibiotics and painkillers, EU standards for maximum amounts in foodstuffs apply.

The EU enforces strict controls: food products such as meat, milk, and eggs are tested for these substances. Because the use of prohibited growth-stimulating substances can often be detected on the farm, urine, feces, hair, serum, wipe samples, etc., are also tested. WFSR is the reference laboratory for substances in Groups A and B (see Annex I of CDR (EU) 2022/1644). WFSR is an expert in analyzing residues of these substances, using both instrumental methods and biosensors and on-site tests. WFSR collaborates closely with EU reference laboratories, coordinates ring tests to improve analytical methods, and provides scientific and technical support to the competent authority.

WFSR has extensive knowledge and experience with the analysis of pesticides in (animal) feed, related legislation, method development, validation, quality control, and the assurance of Official Laboratories (OLs): laboratories that analyze samples within the framework of official EU and national control and monitoring programs.

As the NRL, we advise the Dutch government on technical matters related to the NRL domain and serve as the link between the four European Reference Laboratories (EURLs) [1] for pesticides and the OLs. We participate annually in workshops organized by the EURLs and in collaborative studies organized by them. This keeps our knowledge up-to-date and guarantees the quality of our analyses. It also ensures that the results from different laboratories in the EU are comparable.

Feed additives are substances, microorganisms, or preparations that are intentionally added to animal feed or water, for example, to improve feed properties.

There are five categories:

• technological additives, such as preservatives

• sensory additives, such as aromas and flavors

• nutritional additives, such as vitamins

• zootechnical additives, such as digestive enzymes

• coccidiostats and histomonostats, for the prevention of coccidiosis and histomoniasis

Authorization of Feed Additives
Feed additives may only be used if they have been granted European authorization. A dossier must be submitted with each application. The rules for registration are set out in Regulation (EC) No. 1907/2008. 1831/2003.

WFSR supports the EURL (European Reference Laboratory) in assessing analytical methods as a rapporteur for dossiers and by providing comments on assessments prepared by other NRLs.

Assessing exemption applications
In addition, WFSR assesses applications for exemptions to (temporarily) use unauthorized additives in experiments for scientific purposes.

Furthermore, WFSR advises the Dutch government on quality and analytical aspects of additives with a view to safety, effectiveness, and verifiability.

Determination of water, protein, and their ratio
The NRL's task with determining the water content in poultry meat includes not only determining the water content but also determining the protein content and calculating the water-protein ratio. This ratio is the basis on which certain batches of chicken meat are approved or rejected to protect consumers and ethical producers from poultry meat with excessive water additions. The methods and requirements are laid down in Regulation (EC) No. 543/2008.

The NRL ensures that the methods are correctly implemented by the official laboratories.

Trend in water content in poultry meat
In addition, the long-term trend in the water-protein ratio in various products is monitored, both for Dutch samples and for non-EU samples imported through Dutch ports. Research is also being conducted to determine whether the applicable regulations are sufficient to detect unfair water addition to meat.

WFSR is the national center of expertise in this domain and conducts both analyses and scientific research in virology.

Norovirus and Hepatitis A Virus
The NRL collaborates with other European NRLs and the EURL (European Reference Laboratory) in Sweden. The main goal of this network is to harmonize methods for the official control of food for viral contamination. Norovirus, hepatitis A virus, and hepatitis E virus are the most important viruses that pose a risk to public health via food. ISO 15216 is used to detect and quantify norovirus and hepatitis A virus RNA in food. The NRL network also continuously works on methods not described in ISO 15216.

Food improvement agents are used to improve various properties of food products. They can be divided into three main categories: Additives, Flavorings, and Enzymes. Additives help extend shelf life and maintain quality during production, packaging, and storage. Flavorings preserve, modify, or improve the aroma or taste of foods. Enzymes have a specific biochemical effect and are used for technological purposes. Food improvers are permitted at the European level, and legal limits have been established to ensure safe food consumption.

WFSR is the National Reference Laboratory (NRL) for food improvement agents. We advise the Dutch government on technical matters related to the NRL domain and act as a link between the European Reference Laboratory and official laboratories. Participation in workshops and proficiency tests organized by the EURL keeps our knowledge up-to-date and ensures the quality of our analyses and the compar