In vitro toxicology

From Wikipedia, the free encyclopedia

https://en.wikipedia.org/wiki/In_vitro_toxicology 

In vitro toxicity testing is the scientific analysis of the toxic effects of chemical substances on cultured bacteria or mammalian cells. In vitro (literally 'in glass') testing methods are employed primarily to identify potentially hazardous chemicals and/or to confirm the lack of certain toxic properties in the early stages of the development of potentially useful new substances such as therapeutic drugs, agricultural chemicals and food additives.

In vitro assays for xenobiotic toxicity are recently carefully considered by key government agencies (e.g., EPA; NIEHS/NTP; FDA), to better assess human risks. There are substantial activities in using in vitro systems to advance mechanistic understanding of toxicant activities, and the use of human cells and tissue to define human-specific toxic effects.

Improvement over animal testing

Most toxicologists believe that in vitro toxicity testing methods can be more useful, more time and cost-effective than toxicology studies in living animals (which are termed in vivo or "in life" methods). However, the extrapolation from in vitro to in vivo requires some careful consideration and is an active research area.

Due to regulatory constraints and ethical considerations, the quest for alternatives to animal testing has gained a new momentum. In many cases the in vitro tests are better than animal tests because they can be used to develop safer products.

The United States Environmental Protection Agency studied 1,065 chemical and drug substances in their ToxCast program (part of the CompTox Chemicals Dashboard) using in silica modelling and a human pluripotent stem cell-based assay to predict in vivo developmental intoxicants based on changes in cellular metabolism following chemical exposure. Major findings from the analysis of this ToxCast_STM dataset published in 2020 include: (1) 19% of 1065 chemicals yielded a prediction of developmental toxicity, (2) assay performance reached 79%–82% accuracy with high specificity (> 84%) but modest sensitivity (< 67%) when compared with in vivo animal models of human prenatal developmental toxicity, (3) sensitivity improved as more stringent weights of evidence requirements were applied to the animal studies, and (4) statistical analysis of the most potent chemical hits on specific biochemical targets in ToxCast revealed positive and negative associations with the STM response, providing insights into the mechanistic underpinnings of the targeted endpoint and its biological domain.

A 96-well microtiter plate being used for ELISA.

Examples of cell viability and other cytotoxicity assays used for in-vitro toxicology

Many methods of analysis exist for assaying test substances for cytotoxicity and other cellular responses.

Hemolysis assay

The hemolysis assay examines the propensity of chemicals, drugs or medication, or any blood-contacting medical device or material to lyse red blood cells (erythrocytes). The lysis is easily detected due to the release of hemoglobin.

MTT

MTT assay is used often in determining cell viability and has been validated for use by international organisations. MTT assay involves two steps of introducing the assay to the chemicals and then a solubilisation step.

MTS

The colorimetric MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2Htetrazolium) in vitro assay is an updated version of the validated MTT method, MTS assay has the advantage of being soluble. Hence, no solubilisation step is required.

ATP

ATP assay has the main advantage of providing results quickly (within 15 minutes) and only requires fewer sample cells. The assay performs lysis on the cells and the following chemical reaction between the assay and ATP content of cells produces luminescence. The amount of luminescence is then measured by a photometer and can be translated into number cells alive since

• ATP assay assumes alive cells still have ATP inside them, and
• Luminescence level recorded is proportional to the ATP content in the sample cells.

Neutral Red

Another cell viability endpoint can be Neutral Red (NR) uptake. Neutral Red, a weak cationic dye penetrates cellular membranes by non-diffusion and accumulates intercellularly in lysosomes. Viable cells take up the NR dye, damaged or dead cells do not.

Enzyme-linked immunosorbent assay (ELISA)

ELISA kits can be used to examine up and down regulation of proinflammatory mediators such as cytokines (IL-1, TNF alpha, PGE2)....

Measurement of these types of cellular responses can be windows into the interaction of the test article on the test models (monolayer cell cultures, 3D tissue models, tissue explants).

Types of in vitro studies

Broadly speaking, there are two different types of in vitro studies depending on the type system developed to perform the experiment. The two types of systems generally used are : a) Static well plate system and b) the multi-compartmental perfused systems.

Static well plate system

The static well plate or layer systems are the most traditional and simplest form of assays widely used for in vitro study. These assays are quite beneficial as they are quite simple and provide a very accessible testing environment for monitoring chemicals in the culture medium as well as in the cell. However the disadvantage of using these simple static well plate assays is that, they cannot represent the cellular interactions and physiologic fluid flow conditions taking place inside the body.

Multi-compartmental perfused systems

New testing platforms are now developed to solve problems related to cellular interactions. These new platforms are much more complex based on multi-compartmental perfused systems. The main objective of these systems is to reproduce in vivo mechanisms more reliably by providing cell culture environment close to the in vivo situation. Each compartment in the system represent a specific organ of the living organism and thus each compartment has a specific characteristics and criteria. Each compartment in these systems are connected by tubes and pumps through which the fluid flows thus mimicking the blood flow in the in vivo situation. The draw back behind the use of these perfused systems is that, the adverse effects ( influence of both the biological and non-biological components of the system on the fate of the chemical under study) are more compared to the static systems. In order to reduce the effect of non-biological components of the system, all the compartments are made of glass and the connecting tubes are made up of teflon. A number of kinetic models have been proposed to take care of these non-specific bindings taking place in these in vitro systems.

To improve the biological difficulties arising from the use of different culture in vitro conditions, the traditional models used in flasks or micro-well plates has to be modified. With parallel development in micro-technologies and tissue engineering, these problems are solved using new pertinent tools called "micro-fluidic biochips."

Animal products in pharmaceuticals

From Wikipedia, the free encyclopedia

https://en.wikipedia.org/wiki/Animal_products_in_pharmaceuticals

Animal products in pharmaceuticals play a role as both active and inactive ingredients, the latter including binders, carriers, stabilizers, fillers, and colorants. Animals and their products may also be used in pharmaceutical production without being included in the product itself.

The religious, cultural, and ethical concerns of patients and the disclosure of animal ingredients in pharmaceuticals are a growing area of concern for some people. These would include people who abide by veganism ("vegans"), the practice of abstaining from the use of animal products. Vegan medicines are medications and dietary supplements that do not have any ingredients of animal origin. The vegan status can be determined either through self-proclamation of the company or certification from a third-party organization, such as the Vegan Society or PETA.

Desire for ingredient information

There is public interest in knowing whether medications and supplements contain animal-sourced ingredients. In a study of 100 people, 84% reported not knowing that several medications contained ingredients derived from animal sources. Nearly 63% of the people wanted their physicians, and 35% of the people wanted other healthcare providers (pharmacists, nurses), to notify them when using such medications. Alternatives exist for many animal-derived ingredients, and healthcare providers are increasingly incorporating awareness around animal-free drugs in their medical practice.

A 2013 study in the BMC Medical Ethics contacted branches of six of the world's largest religions. Of the six religions contacted, respondents from three did not accept or approve of the use of animal products in pharmaceuticals. The authors concluded that:

...religious codes conflict with some treatment regimens. It is crucial to obtain informed consent from patients for the use of drugs and implants with animal or human derived content. However, information on the origin of ingredients in drugs is not always available to health practitioners.

Similarly, a 2014 BMJ analysis on the topic discussed the lack of information about ingredients available to doctors. According to the article, "Most medications prescribed in primary care contain animal derived products" and "Disclosure of animal content and excipients would help patients make an informed personal choice"

Active ingredients in drugs and dietary supplements

Biomedicine

• Insulin from cattle and pigs has been used since the 1920s, and was the predominant form of insulin used for decades. The first synthetic human insulin was created using bacteria in 1978. In the United States, the manufacture of beef insulin was discontinued in 1998, and the manufacture of pork insulin was discontinued in 2006.
• Premarin, a hormone replacement therapy, is a conjugated estrogen. It was first available in the form of a preparation manufactured from the urine of pregnant mares - hence "Premarin" from "PREgnant MARe's urINe". It is now also made as a fully synthetic product.

Dietary supplements

• Glucosamine, used in dietary supplements marketed for osteoarthritis, is extracted from chitin from shellfish. Non-animal sourced glucosamine is also available.
• Cartilage as a dietary supplement is by definition animal-sourced. Shark cartilage is marketed explicitly or implicitly as a treatment or preventive for various illnesses, including cancer. There is no consensus that shark cartilage is useful in treating or preventing cancer or other diseases.

Traditional Chinese Medicine

Traditional Chinese Medicine (TCM) utilizes approximately 1,000 plant species and 36 animal species. Animal ingredients in TCM include animal parts such as tiger bones, rhino horns, deer antlers, and snake bile. The use of animal parts in TCM have been definitively linked to the extinction of wildlife. One example of this link is the pangolin trade, which has led the pangolin to be called the world's "most trafficked mammal." In 2020, pangolin scales were removed from the Chinese list of ingredients approved for use in Traditional Chinese Medicine.

Homeopathic medicine

Blatta Orientalis

Homeopathic medicine is made of plants, minerals, or animal parts. Oscillococcinum, a remedy purported to reduce cold and flu like symptoms, is made of duck heart and liver. There is also use of insects in homeopathic medicine, such as Blatta orientalis, a type of cockroach which has been studied by homeopaths for anti-asthmatic effects.

Inactive ingredients

• Gelatin is derived from animal skin, bone, and tissue most often from pigs or beef. There is no practical way of determining if the gelatin used in pharmaceuticals is derived from beef or pork. It is used primarily for gel capsules and as stabilizers for vaccines. Non-animal derived alternatives to gelatin include pectin as a gelling agent or cellulose for creating capsules.
• Lactose is derived from cow's milk and is a frequently used filler or binder in tablets and capsules.
• Magnesium stearate is the most commonly used emulsifier, binder, thickener, or lubricant. It can be derived from animal- or plant-sourced stearic acid, although it is most commonly sourced from cottonseed oil or palm oil.
• Sodium tallowate is a common soap ingredient derived from tallow—the fat of animals such as cattle and sheep. A popular alternative to this ingredient is sodium palmate, which is derived from palm oil. Soap is a pharmaceutical according to the United States Food and Drug Administration.
• Shellac is a resin excreted by female insects of the species Kerria lacca. It is used as a glazing agent on pills.
• Carmine, derived from crushed cochineal beetles, is a red or purple substance commonly used in pharmaceutical products. Evidence shows that it can be allergenic. Carmine is an allergen according to the US Food and Drug Administration (FDA). The FDA requires this ingredient to be declared in food and cosmetics, but not pharmaceuticals.

Animal use during product development or production

Main articles: Animal testing and Alternatives to animal testing

A separate issue is the use of testing on animals as a means of initial testing during drug development, or actual production. Guiding principles for more ethical use of animals in testing are the Three Rs first described by Russell and Burch in 1959. These principles are now followed in many testing establishments worldwide.

1. Replacement refers to the preferred use of non-animal methods over animal methods whenever it is possible to achieve the same scientific aim.
2. Reduction refers to methods that enable researchers to obtain comparable levels of information from fewer animals, or to obtain more information from the same number of animals.
3. Refinement refers to methods that alleviate or minimize potential pain, suffering, or distress, and enhance animal welfare for the animals used.

Cow blood is used in vaccine manufacture. Microorganisms for vaccine manufacture are grown under controlled conditions in liquid solutions ("media") which provide the nutrients necessary for growth. These can include cow plasma. Chicken eggs are used in the production process of some vaccines. For influenza vaccination there are non-egg alternatives.

Alternatives to animal testing

From Wikipedia, the free encyclopedia

https://en.wikipedia.org/wiki/Alternatives_to_animal_testing 

 

Animal testing
Main articles
Animal testing Alternatives to animal testing Animal testing regulations History of animal testing History of model organisms IACUC Laboratory animal sources Pain and suffering in laboratory animals Testing cosmetics on animals Toxicology testing Vivisection
Testing on
Invertebrates Frogs Primates Rabbits Rodents
Issues
Medical research Animal rights, Animal welfare Animals (Scientific Procedures) Great ape research ban International primate trade
Cases
Brown Dog affair Cambridge University primates Pit of despair Silver Spring monkeys UCR 1985 laboratory raid Unnecessary Fuss
Companies
Jackson Laboratory Charles River Laboratories Covance, Harlan Huntingdon Life Sciences UK lab animal suppliers Nafovanny, Shamrock Farm
Groups/campaigns
AALAS, AAAS, ALF Americans for Medical Progress Boyd Group, Cruelty Free International Dr. Hadwen Trust Foundation for Biomedical Research FRAME National Anti-Vivisection Society New England Anti-Vivisection Society PETA, Physicians Committee for Responsible Medicine Primate Freedom Project Pro-Test SPEAK campaign, SHAC Speaking of Research Understanding Animal Research
Writers/activists
Tipu Aziz Michael Balls Neal D. Barnard Colin Blakemore Simon Festing Gill Langley Ingrid Newkirk Bernard Rollin Jerry Vlasak Syed Ziaur Rahman
Categories
Animal testing Animal rights Animal welfare

Alternatives to animal testing are the development and implementation of test methods that avoid the use of live animals.

There is widespread agreement that a reduction in the number of animals used and the refinement of testing to reduce suffering should be important goals for the industries involved. Two major alternatives to in vivo animal testing are in vitro cell culture techniques and in silico computer simulation. However, some claim they are not true alternatives because simulations use data from prior animal experiments and cell cultures often require animal derived products, such as serum or cells. Others say that they cannot replace animals completely as they are unlikely to ever provide enough information about the complex interactions of living systems. Other alternatives include the use of humans for skin irritancy tests and donated human blood for pyrogenicity studies. Another alternative is so-called microdosing, in which the basic behaviour of drugs is assessed using human volunteers receiving doses well below those expected to produce whole-body effects. While microdosing produces important information about pharmacokinetics and pharmacodynamics it does not reveal information about toxicity or toxicology. Furthermore, it was noted by the Fund for the Replacement of Animals in Medical Experiments that despite the use of microdosing, "animal studies will still be required".

Guiding principles for more ethical use of animals in testing are the Three Rs (3Rs) first described by Russell and Burch in 1959. These principles are now followed in many testing establishments worldwide.

1. Replacement refers to the preferred use of non-animal methods over animal methods whenever it is possible to achieve the same scientific aim.
2. Reduction refers to methods that enable researchers to obtain comparable levels of information from fewer animals, or to obtain more information from the same number of animals.
3. Refinement refers to methods that alleviate or minimize potential pain, suffering, or distress, and enhance animal welfare for the animals used.

Cell culture and tissue engineering

Cell culture in a special tissue culture dish

Cell culture can be an alternative to animal use in some cases. For example, cultured cells have been developed to create monoclonal antibodies; prior to this, production required animals to undergo a procedure likely to cause pain and distress. However, even though cell or tissue culture methods may reduce the number of experiments performed on intact animals, the maintenance of cells in culture normally requires the use of animal-derived serum. Although exact figures are difficult to obtain, some have estimated that one million foetal cows are sacrificed each year to obtain the world's supply of foetal bovine serum, used to grow cultured cells. The testing of cosmetic products directly onto an animal can be minimized or eliminated by the use of in vitro cell growth and development. This can be generalized as the growth of cells outside of the body and tested on without causing harm or pain on the test subject. Much of the time this method of cosmetic testing is less time-consuming and less expensive than alternative choices.

Skin corrosion and skin irritation

Skin irritation and skin corrosion refer to localized toxic effects resulting from a topical exposure of the skin to a substance. Human skin equivalent tests can be used to replace animal-based corrosive and irritative studies. EpiDerm from Mattek and EpiSkin and SkinEthic RHE model are derived from human skin cells which have been cultured to produce a model of human skin. These methods are currently accepted replacements in Canada and the European Union (EU). In August 2010, the Organisation for Economic Co-operation and Development (OECD) published the Test Guideline 439 which describes the new procedure for in vitro hazard identification of irritant chemicals.

Another synthetic replacement uses a protein membrane to simulate a skin barrier and is approved as a partial replacement by the US Department of Transportation and European Union.

Skin absorption

Several tissue culture methods that measure the rate of chemical absorption by the skin have been approved by the OECD.

Phototoxicity

Phototoxicity is a rash, swelling, or inflammation, like a severe sunburn, caused by exposure to light following exposure to a chemical. The 3T3 Neutral Red Uptake (NRU) Phototoxicity Test, approved by the OECD, detects the viability of 3T3 cells after exposure to a chemical in the presence or absence of light. The 3T3 cell line was developed in 1962 and is derived from mouse embryonic fibroblast cells.

Fungal model for mammalian drug metabolism

Fungi like Cunninghamella elegans can be used as a microbial model of mammalian drug metabolism thereby reducing the need for laboratory animals.

Prokaryotes are often used as an alternative to animal testing. Prokaryotes include bacteria such as Escherichia coli (E. coli) or Bacillus subtilis. These bacteria are the ideal model for genetic and molecular studies. Fungi is also used as an alternative for animal testing. Certain fungi can be used for genetic studies or circadian rhythms studies. This may include Neurospora crassa, otherwise known as a type of red mould. Invertebrates are another ideal candidate for testing. One of the most common invertebrates tested on include Drosophila melanogaster, the fruit fly. Fruit flies are used to find human diseases.

Organoids (3D cell cultures)

Russell and Burch writing six decades ago could not have anticipated some of the technologies that have emerged today. One of these technologies, 3D cell cultures, also known as organoids or mini-organs, have replaced animal models for some types of research. In recent years, scientists have produced organoids that can be used to model disease and test new drugs. Organoids grow in vitro on scaffolds (biological or synthetic hydrogels such as Matrigel) or in a culture medium. Organoids are derived from three kinds of human or animal stem cells—embryonic pluripotent stem cells (ESCs), adult somatic stem cells (ASCs), and induced pluripotent stem cells (iPSCs). These organoids are grown in vitro and mimic the structure and function of different organs such as the brain, liver, lung, kidney, and intestine. Organoids have been developed to study infectious disease. Scientists at Johns Hopkins University have developed mini-brain organoids to model how COVID-19 can affect the brain. Researchers have used brain organoids to model how the Zika virus disrupts foetal brain development. Tumoroids—3D cell cultures derived from cells biopsied from human patients—can be used in studying the genomics and drug resistance of tumours in different organs. Organoids are also used in modelling genetic diseases such as cystic fibrosis, neurodegenerative diseases such as Alzheimer's and Parkinson's, infectious diseases such as MERS-CoV and norovirus, and parasitic infections such as Toxoplasma gondii. Human- and animal-cell-derived organoids are also used extensively in pharmacological and toxicological research.

Human-based

Skin irritation

A skinpatch test has been designed and is used in Canada to measure development of rashes, inflammation, swelling or abnormal tissue growth on human volunteers. Unlike corrosives, substances defined as irritants cause only reversible skin damage.

Another approach has been the development of test methods that use cultured human cells. Human epidermal keratinocytes have been cultured to mimic the human epidermis, and are used to measure skin irritation and dermal corrosion. This method has been accepted by the EU and is intended to replace the Draize rabbit skin irritation test.

Pyrogenicity

Pyrogens are most often pharmaceutical products or intravenous drugs that may cause inflammation or fever when they interact with immune system cells. This interaction can be quickly and accurately tested in vitro.

Modular immune in vitro construct

The modular immune in vitro construct (MIMIC) uses human cells to create a model of the human immune system on which the efficacy of new vaccines and other compounds may be tested, replacing some steps of the vaccine development process that would otherwise be performed on animals. This process is faster and more flexible than previous methods but critics worry that it may be too simple to be useful on a large scale.

Medical imaging

Further information: Medical imaging and Microdosing

Medical imaging is able to demonstrate to researchers both how drugs are metabolised by use of microdosing, and the detailed condition of organ tissue.

Computer simulation

Crash test dummies have been used to replace live animals in impact testing.

 

See also: In silico

Examples of computer simulations available include models of asthma, though potential new medicines identified using these techniques are currently still required to be verified in animal and human tests before licensing.

Computer operated mannequins, also known as crash test dummies, complete with internal sensors and video, have replaced live animal trauma testing for automobile crash testing. The first of these was "Sierra Sam" built in 1949 by Alderson Research Labs (ARL) Sierra Engineering. These dummies continue to be refined. Prior to this, live pigs were used as test subjects for crash testing.

Computer models have been constructed to model human metabolism, to study plaque build-up and cardiovascular risk, and to evaluate toxicity of drugs, tasks for which animals are also used. In 2007, US researchers using the world's fastest computer at the time, BlueGene L, modelled half a mouse brain for just 10 seconds. However, due to limitations in computing power, the simulation could only be run at one-tenth the speed of an actual mouse brain. Although this was an advance in science, its representative power as a model was limited and the researchers were quoted as saying that "although the simulation shared some similarities with a mouse's mental make-up in terms of nerves and connections it lacked the structures seen in real mice brains."

In pharmacology and toxicology, physiologically based pharmacokinetic models can be used for in vitro to in vivo extrapolation and to predict the time-dependent distribution of chemicals in the organism, while quantitative structure-activity relationship (QSAR) models can be used to predict the physicochemical and hazard properties of chemicals.

Microfluidic chips

Further information: Microfluidics and Lab-on-a-chip

Microfluidic chips, which are just 2 cm (0.79 in) wide, can be engraved into a series of small chambers, each containing a sample of tissue from a different part of the body. A substitute of blood flows through micro-channels where the compartments of chips linked. When injected, the test drug circulates around the device, mimicking what goes in the body on a micro scale. Sensors in the chip transfer information for computer analysis.

Another name for this chip is the microfluidic chip is cell-bio chips. With the capacity to "perform perfusion culture" and reproduce "physiological conditions such three-dimensional architectures, circulatory flowrate and zonation and multi cellular co-cultures", the biochips have set themselves apart from basic cell cultures analysed in a Petri dish. The effectiveness of these systems is constantly being increased with various new materials that can be used to make it. An ideal material would be gas permeable but still be able to absorb molecules that would be expected to be found in various drugs.

The choice of the material for chips is still challenging. One of the major materials that can be possibly used in chips is known as polydimethylsiloxane (PDMS). However, due to lack of facilities for mass production and drug clearance issue, the use of PDMS is still being speculated, even though it has great properties as microfluidic chip. Also, the biological process involved in proliferation and metabolism might be modified when compared to larger scales, because the materials have micro-structured scales comparable in size to cells.

Future alternatives

Organs on a chip

Main article: Organ-on-a-chip

The Wyss Institute for Biologically Inspired Engineering (US) intends to develop in-vitro organs for drug screening and thereby eliminate the use of animals for this type of testing. One model is the "lung-on-a-chip". This combines microfabrication techniques with modern tissue engineering and mimics the complicated mechanical and biochemical behaviours of a human lung.

Human toxome

Toxicity testing typically involves studying adverse health outcomes in animals subjected to high doses of toxicants with subsequent extrapolation to expected human responses at lower doses. The system relies on the use of a 40+year-old patchwork of animal tests that are expensive (costing more than $3B per year), time-consuming, low-throughput and often provide results of limited predictive value for human health effects. The low-throughput of current toxicity testing approaches (which are largely the same for industrial chemicals, pesticides and drugs) has led to a backlog of more than 80,000 chemicals to which humans are potentially exposed whose potential toxicity remains largely unknown. In 2007, the National Research Council (NRC) released the report "Toxicity Testing in the 21st Century: A Vision and a Strategy", that charted a long-range strategic plan for transforming toxicity testing. The major components of the plan include the use of predictive, high-throughput cell-based assays (of human origin) to evaluate perturbations in key toxicity pathways, and to conduct targeted testing against those pathways. This approach will greatly accelerate our ability to test the vast "storehouses" of chemical compounds using a rational, risk-based approach to chemical prioritization, and provide test results that are hopefully far more predictive of human toxicity than current methods. Although a number of toxicity pathways have already been identified, most are only partially known and no common annotation exists. Mapping the entirety of these pathways (i.e. the Human Toxome) will be a large-scale effort, perhaps on the order of the Human Genome Project.

Research initiatives

SEURAT-1

SEURAT-1 is a long-term strategic target for "Safety Evaluation Ultimately Replacing Animal Testing". It is called "SEURAT-1" to indicate that more steps have to be taken before the final goal will be reached. SEURAT-1 will develop knowledge and technology building blocks required for the development of solutions for the replacement of current repeated dose systemic toxicity testing in vivo used for the assessment of human safety. SEURAT-1 is composed of six research projects, which started on January 1, 2011 and will run for five years. These projects will closely cooperate with a common goal and combine the research efforts of over 70 European universities, public research institutes and companies. The collaboration between these six research projects, the dissemination of results, the cooperation with other international research teams, and the continuous updating on research priorities will be facilitated by the coordination and support action project "COACH".

SEURAT-1 was developed through the Framework Programme 7 (FP7) research initiative and was created through a call for proposals by the European Commission (EC) that was published in June 2009. The Cosmetics Europe industry offered to match the EC's funds to make a total of EUR 50 million available to try to fill current gaps in scientific knowledge and accelerate the development of non-animal test methods.

Euroecotox

Laboratory animals are not restricted to rats, mice, dogs, and rabbits, but also include fish, frogs and birds. Research into alternatives to replace these species is often neglected, although fish are the third most widely used laboratory animal used for scientific purposes in the EU. This is also the field where until now only two alternative tests exist worldwide: One guideline, OECD TG 236, and one guidance (OECD series on testing and assessment 126) are so far available.

Euroecotox is a European network for alternative testing strategies in ecotoxicology. It was funded by the Seventh Framework Programme (FP7) of the European Commission Environment Programme. The main objectives of the Euroecotox network are: To contribute to the advancement of alternative methods of ecotoxicity testing in Europe. To promote the validation and regulatory acceptance of new alternative ecotoxicity methods. To facilitate the networking of research groups working in the field of alternative ecotoxicology. To provide a gathering point for all stakeholders involved in the development, validation, regulatory acceptance and final use of alternative ecotoxicity testing strategies. To act as the one voice for alternative ecotoxicity testing in Europe.

AXLR8

AXLR8 is a coordination action funded by the European Commission Directorate General for Research & Innovation under the 7 Framework Programme 7 (FP7) Health Theme. The European Commission is currently funding a number of research consortia to develop new 3Rs (replacement, reduction and refinement) test methods and strategies as potential alternatives to the use of animals in safety testing. Monitoring of these 3Rs activities at pan-European, national, and international levels is vital to facilitate swift progress. AXLR8 aims to fulfil this growing need by providing a focal point for dialogue and collaboration. Humane Society International is part of the consortium.

Regulation

European Union

EU Directive 2010/63/EU

Main article: EU Directive 2010/63/EU

On January 1, 2013, EU Directive 2010/63/EU "on the protection of animals used for scientific purposes" entered into force for the EU member states (MS), repealing Directive 86/609/EEC. Because it is a directive, it allows member states certain flexibility in transposition of national rules. The status of the implementation of the new directive in the EU is described by the EC General Environment Directorate.

Article 1.3: The new EU directive applies to the following animals: (a) live non-human vertebrate animals, including: (i) independently feeding larval forms; and (ii) foetal forms of mammals from the last third of their normal development; (b) live cephalopods.

Article 4: The directive refers directly to the 3Rs: "Principle of replacement, reduction and refinement".

Article 47-2: Member states shall assist the commission in identifying and nominating suitable specialised and qualified laboratories to carry out such validation studies.

In July 2013, the commission announced the creation of NETVAL (European Union Network of Laboratories for the Validation of Alternative Methods). EU-NETVAL's primary role is to provide support for EURL ECVAM validation projects, including aspects of training and dissemination, and the identification of methods that have a potential to reduce, refine or replace animals used for scientific purposes. Currently there are thirteen test facilities in nine member states: Germany (3), the Netherlands (2), Spain (2), Belgium (1), Czech Republic (1), Finland (1), France (1), Italy (1) and Sweden (1).

Other regulations

The Cosmetics Directive provides the regulatory framework for the phasing out of animal testing for cosmetics purposes. It establishes prohibitions against (a) testing finished cosmetic products and cosmetic ingredients on animals (testing ban), and (b) marketing in the EU finished cosmetic products and ingredients included in cosmetic products which were tested on animals for cosmetics purposes (marketing ban). The same provisions are contained in Cosmetics Regulation EU 1223/2009, which replaces the Cosmetics Directive as of July 11, 2013.

In 2007, EU legislation on the Registration, Evaluation, Authorisation and Restriction of Chemicals (REACH EC 1907/2006) came into force, relating to chemicals and their safe use. The aim of REACH is to improve the protection of human health and the environment through the better and earlier identification of the intrinsic properties of chemical substances. It promotes the use of alternative methods for animal testing, but does not oblige the test performer to do so; "Article 25.1 - In order to avoid animal testing, testing on vertebrate animals for the purposes of this Regulation shall be undertaken only as a last resort. It is also necessary to take measures limiting duplication of other tests."

In parallel to the adoption of REACH, the EC published standardised and accepted methods for testing hazardous properties of chemicals. These were written into the "Test Methods Regulation". All the alternative test methods among the in vivo studies are included in PART B; "The European Union is committed to promoting the development and validation of alternative techniques which can provide the same level of information as current animal tests, but which use fewer animals, cause less suffering or avoid the use of animals completely. Such methods, as they become available, must be considered wherever possible for hazard characterisation and consequent classification and labeling for intrinsic hazards and chemical safety assessment."

EU philosophy on food additives, food enzymes, and food flavourings and ingredients intended for human consumption is that none should be put on the market unless they are included on a published Community list of authorised substances, in accordance with the conditions laid down in relevant food law. This approach is intended to bring food producers into compliance with the provisions of Regulation (EC) 1334/2008 that pertain to the safety of food flavourings. As part of the approval process, the EC will require full disclosure of study data, safety issues, and toxicological findings for all such additives.

Within the EU animal welfare law (2010/63/EU), the principles of the 3Rs are invoked whenever toxicological test methods are necessary.

Organizations and programs

Scientific congresses

The European Society for Alternatives to Animal Testing (EUSAAT) organises an annual conference in Linz (Austria) for

1. Dissemination and validation of alternative methods to animal testing
2. Promotion of research in the field of the 3Rs
3. Reduction of the use of animals for tests in the field of education and continuing education
4. Reduction of suffering and stress of laboratory animals by better breeding, keeping, test planning and other accompanying measures
5. Experts' guidance and referees' opinion for public and private organizations, companies, universities
6. Suitable information for the public and the media

The World Congress on Alternatives and Animal Use in the Life Sciences takes place every three years. The next conference (10th) will be held in September 2017 in Seattle.

The 1st Latino-Americano Congress on Alternative to Animal Testing took place in 2012. Colama (I Congresso Latino-Americano De Metodos Alternativos Ao Uso De Animais No Ensino, Pesquisa E Industria).

The Johns Hopkins University Center for Alternatives to Animal Testing (CAAT) co-organizes an annual symposium on the 3Rs with the USDA's Animal Welfare Information Center (AWIC) and NIH's Office of Laboratory Animal Welfare. Previously known as the Social Housing Symposium, the symposium has occurred annually (except for 2015) since 2013 with past symposia archived on video on the AWIC website. Videos of the most recent symposium, "7th Annual 3Rs Symposium: Practical Solutions and Success Stories", held in June 2020, may also be found on the AWIC website.

Industry and corporate initiatives

• Cosmetics Europe: Represents the interests of more than 4000 companies in the cosmetic, toiletry and perfumery industry since 1962.
• Unilever: "We do not test finished products on animals unless demanded by the regulatory authorities in the few countries where this is the law. In such cases, we try to convince the local authorities to change the law. Where some testing of ingredients is required by law or currently unavoidable, we aim to minimise the number of animals used."
• BASF: "Systematic screening investigations provide information about important toxicological properties of substances at an early stage of development. ... We replace animal experiments whenever an alternative method is available that complies with an OECD Test Guideline and is recognized by the authorities."

Animal welfare and animal rights organizations

Further information: Animal rights and List of animal welfare groups

• Eurogroup for Animals: "An estimated 12.1 million animals – including dogs, rabbits and even our closest genetic relatives, primates – are used in laboratory research throughout Europe every year. Eurogroup focusses on ensuring their protection and works with legislators, experts and industry with the aim of ultimately replacing all animal experiments with viable alternatives. We continue to actively promote the replacement, reduction and refinement of animal tests and do all we can to improve the lives of those animals currently used for research."
• Vier Pfoten (Four Paws) (Austria)
• Antidote (France) "When it is about assessing drug safety, humans are not 70kg rats! It is about time to move on from the actual paradigm on assessing drug safety. The first step would be to eliminate all the regulatory requirements for animal testing and replace these tests by 21st century methods."
• Deutscher Tierschutzbund (Germany)
• Lega Anti Vivisezione (Italy)
• The ALEXANDRA Association (Monaco): "... aims at stimulating research and development (R&D) in the area of alternative methods to animal experimentation by providing political, technical and educational support to researchers and entrepreneurs worldwide. In particular alternative methods based on 'Open Source' concepts i.e. non-patent protected core technologies for human tissue reconstruction and cell culture technologies will be actively promoted."
• British Union for the Abolition of Vivisection (BUAV): "For over 100 years the BUAV has been campaigning peacefully to create a world where nobody wants or believes we need to experiment on animals."
• New England Anti-Vivisection Society (NEAVS) (United States): "Recognition of the inadequacy of animal toxicity testing has resulted in the development of better techniques ... NEAVS and its programs will help hasten the inevitable and necessary transition away from animal-based experimentation, testing, and teaching, toward science and science education governed by progressive scientific thought and compassionate ethics."
• Humane Society International (HSI) in the US and UK: "Today, scientific and government authorities worldwide are acknowledging the deficiencies of "animal models" and calling for a new approach to safety testing and health research using state-of-the-art techniques. Advances in biology, genetics, computer science and robotics have given scientists new tools to help identify the root causes of human toxicity and disease."
• People for the Ethical Treatment of Animals (PETA) in the US and UK: "We teamed up with CeeTox, Inc. to fund work on a new humane skin test that could replace painful tests on mice and guinea pigs."

Public campaigns and awards

• Petition to the European Parliament for the abolition of vivisection as a European Citizen Initiative. The threshold of 1 million signatures was reached for the deadline (1 November 2013). The European Commission is currently checking the authenticity of each signature.
• "Go cruelty free" The launch of the global "Go Cruelty-Free" campaign occurred in 2012 and every year since the launch there have been more and more contributions from around the globe to help put a stop to using animals for lab testing. The most recent contribution is from Australia in 2019, they banned the use of newly derived animal test data for cosmetics.
• HSI's report "Advancing Safety Science and Health Research with Innovative, Non-Animal Tools."
• The Lush Prize: "The Lush Prize is a major initiative which will use resources to bring forward the day when safety testing takes place without the use of animals. The Lush Prize will focus pressure on toxicity testing for consumer products and ingredients in a way which complements the many projects already addressing the use of animals in medical testing."
• EPAA (European Partnership for Alternative Approaches to Animal Testing) will grant a €3000 prize to a laboratory technician involved in implementing and raising awareness of Replacement, Reduction and Refinement of animal testing.
• The Alternatives Research and Development Foundation (ARDF) provides grants to advance the use of non-animal methods in the fields of biomedical testing, research, and education.
• The international NC3Rs 3Rs Prize is awarded to highlight an outstanding original contribution to scientific and technological advances in the 3Rs in medical, biological or veterinary sciences published within the last three years.
• The American Fund for Alternatives to Animal Research (AFAAR) funds a wide and encompassing range of research involving the use, development, or validation of alternatives.

Education and training

• IIVS: The Institute for In Vitro Sciences, Inc. is a non-profit research and testing laboratory dedicated to the advancement of in vitro (non-animal) methods worldwide. Founded in 1997, IIVS has worked with industry and government agencies to implement in vitro testing strategies that limit animal use while supplying key information for product safety and efficacy decisions.
• NORINA is a database containing details of products which may be used as alternatives or supplements to the use of animals in education and training. NORINA's search engine is linked to those of two other databases: TextBase, which provides information on textbooks and other written material of relevance to laboratory animal science and alternatives, and 3R Guide which gives details of guidelines, information centres, databases, journals and email lists within the field of replacement, reduction and refinement of animal experimentation. The three databases are hosted by Norecopa.
• InterNICHE is the International Network for Humane Education. It has been developed to meet the needs of teachers and trainers, students, ethics committees, alternatives producers and campaigners internationally.
• "Tierschutz macht Schule" - the Association for Animal Welfare Education - was founded in the course of the implementation of Austria's nationwide animal welfare law. The animal welfare education association aims to improve the living conditions of pets, farm animals, laboratory animals and wild animals through providing knowledge about their needs and behaviour to children, youth and the public. The association offers a teaching magazine about research animals and animal tests suitable for secondary schools and college, which can be ordered on their website. It aims at explaining alternatives to animal testing in a youth appropriate language and can be used in lessons straight away.
• XCellR8's mission is to support, develop and implement the use of scientifically advanced and ethically sound alternatives to animal testing. They are an exclusively in vitro company, with commitment to promoting non-animal testing strategies at the core of all of their activities.
• The Animal Welfare Information Center at the National Agricultural Library (USDA) holds a workshop several times a year called "Meeting the Information Requirements of the Animal Welfare Act". In the workshop, researchers and other stakeholders learn how to perform literature searches for animal use alternatives as well as the history and evolution of the Animal Welfare Act of 1966 and its amendments, showing how the legislation regulates animal welfare.
• EPISKIN Academy is an initiative of the industry to propose training and courses to facilitate the deployment of validated alternative methods to animal testing in toxicology and to prepare next generations of scientists and toxicologists to use these methods. Created in 2012, EPISKIN Academy proposes a modular program ranging from demonstration of these methods to full theoretical and practical laboratory training leading to certification. Based on long-term partnerships with institutional partners in different countries, this program in education allows hands-on training to the methods but also to the scientific and regulatory knowledge important for their implementation and acceptance.

Institutes and national or international organizations

Institutes and organizations that research or fund alternatives to animal testing include:

Asia - Oceania

• Medical Advances Without Animals Trust (Australia)
• Alternatives to Animal Experimentation Laboratory, Department of Pharmacology, Jawaharlal Nehru Medical College, Aligarh Muslim University, Aligarh (India).
• Mahatma Gandhi-Doerenkamp Center for Alternatives to Use of Animals in Life Science, Bharathidasan university, Trichy, Tamil Nadu, India Education
• Japanese Center for the Validation of Alternative Methods (JACVAM), since 2005
• The Korean Center for the Validation of Alternative Methods (KOCVAM), since 2009

South America

• BraCVAM as the Brazilian Center for the Validation of Alternatives Methods. It was established in 2011.

North America

• Canadian Council on Animal Care
• Canadian Centre for Alternatives to Animal Methods, founded in 2017 at the University of Windsor.
• Health Canada, which does not have a formal validation centre, but coordinates health related test method validation and acceptance issues
• American Fund for Alternatives to Animal Research (AFAAR): "Over the years, AFAAR has funded alternatives worldwide, including more than 200 human tissue culture tests to replace toxicity and other testing on animals ... Today, AFAAR funds a wide and encompassing range of research involving the use, development, or validation of alternatives."
• Interagency Coordinating Committee on the Validation of Alternative Methods (ICCVAM) (United States), since 1994;
• Johns Hopkins University Center for Alternatives to Animal Testing
• Physicians Committee for Responsible Medicine (United States)

Europe

• The European Commission's role in promoting the development, validation and uptake of alternative approaches to animal testing started in 1991, with the launch of ECVAM (European Centre for the Validation of Alternative Methods), hosted by the Joint Research Centre. As from 2011, ECVAM became known as the EU Reference Laboratory for Alternatives to Animal Testing (EURL ECVAM). EURL ECVAM hosts an online database of toxicological, non-animal alternative test methods DB-ALM.
• Under the Framework Programmes 6 and 7, the EC funded a significant number of large integrated research projects aiming to develop alternatives to animal testing for about EUR 330 million based on the Review of REACH from February 2013 (the European Chemical Program).
• The European Partnership for Alternative Approaches to Animal Testing (EPAA) as a liaison between the EC and industries.
• The European Consensus Platform for Alternatives (ECOPA) coordinates efforts amongst EU member states.
• Zentrum fuer Ersatz (Austria)
• Finnish Center for Alternative Methods (FICAM), since 2008
• FRANCOPA is the French platform dedicated to development, validation, and dissemination of alternative methods in animal testing. It was created on November 16, 2007.
• The Italian Centro 3R was created in 2017. It is an interuniversity center dedicated to the promotion of the 3Rs in teaching and research.
• Zentralstelle zur Erfassung und Bewertung von Ersatz- und Ergänzungsmethoden (ZEBET) (Germany), since 1989
• Norecopa is the Norwegian consensus platform for the replacement, reduction and refinement of animal experiments. It was founded on October 10, 2007.
• Romanian Center for Alternative Test Methods (ROCAM) promotes the application of alternative methods in industry and their acceptance by regulators in Romania and also the development of new methods and approaches. ROCAM was established in June 2015 with the main goal to support and promote the 3Rs principles in Romania and regionally.
• Dr Hadwen Trust (United Kingdom)
• Fund for the Replacement of Animals in Medical Experiments (United Kingdom)
• National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs) (United Kingdom), since 2004

International

• International Cooperation on Alternative Test Methods (ICATM): On April 27, 2009, the United States, Canada, Japan and EU signed a memorandum of cooperation that could reduce the number of animals required for consumer product safety testing worldwide. The agreement will yield globally coordinated scientific recommendations on alternative toxicity testing methods that should speed their adoption in each of these countries, thus reducing the number of animals needed for product safety testing.

International Cooperation on Alternative Test Methods (ICATM)

 
Legend :
ICH: The International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use
OECD: The Organisation for Economic Co-operation and Development has a Test Guideline program that deals with chemicals.
ICCR: The International Cooperation on Cosmetics Regulation (ICCR) is an international group of cosmetic regulatory authorities from the United States (FDA), Japan (Ministry of Health, Labour, and Welfare), the EU (EC, DG Enterprise), and Canada (Health Canada). This multilateral framework maintains the highest level of global consumer protection, while minimizing barriers to international trade.

• The OECD (Organisation for Economic Co-operation and Development) is a forum for discussion where governments express their points of view, share their experiences, and search for common ground, as opposed to a supranational organization. OECD is a forum where alternative test methods also undergo validation and are therereafter accepted for regulatory purposes in more than 35 member countries worldwide. NGOs are represented at the technical level at the OECD, under the ICOPA International Council on Animal Protection in OECD program. The testing of chemicals is labor-intensive and expensive. Often the same chemical is tested in several countries simultaneously, which means that redundant animal tests are performed. To relieve some of this burden, the OECD Council adopted a decision in 1981, stating that data generated in a member country, in accordance with OECD Test Guidelines and Principles of Good Laboratory Practice (GLP), shall be accepted in other member countries for assessment purposes and other uses relating to the protection of human health and the environment. This principle is referred to using the acronym MAD, for the Mutual Acceptance of Data.

Animal testing on non-human primates

From Wikipedia, the free encyclopedia

https://en.wikipedia.org/wiki/Animal_testing_on_non-human_primates

 

Two primates in a laboratory cage

 

Animal testing
Main articles
Animal testing Alternatives to animal testing Animal testing regulations History of animal testing History of model organisms IACUC Laboratory animal sources Pain and suffering in laboratory animals Testing cosmetics on animals Toxicology testing Vivisection
Testing on
Invertebrates Frogs Primates Rabbits Rodents
Issues
Medical research Animal rights, Animal welfare Animals (Scientific Procedures) Great ape research ban International primate trade
Cases
Brown Dog affair Cambridge University primates Pit of despair Silver Spring monkeys UCR 1985 laboratory raid Unnecessary Fuss
Companies
Jackson Laboratory Charles River Laboratories Covance, Harlan Huntingdon Life Sciences UK lab animal suppliers Nafovanny, Shamrock Farm
Groups/campaigns
AALAS, AAAS, ALF Americans for Medical Progress Boyd Group, Cruelty Free International Dr. Hadwen Trust Foundation for Biomedical Research FRAME National Anti-Vivisection Society New England Anti-Vivisection Society PETA, Physicians Committee for Responsible Medicine Primate Freedom Project Pro-Test SPEAK campaign, SHAC Speaking of Research Understanding Animal Research
Writers/activists
Tipu Aziz Michael Balls Neal D. Barnard Colin Blakemore Simon Festing Gill Langley Ingrid Newkirk Bernard Rollin Jerry Vlasak Syed Ziaur Rahman
Categories
Animal testing Animal rights Animal welfare

Experiments involving non-human primates (NHPs) include toxicity testing for medical and non-medical substances; studies of infectious disease, such as HIV and hepatitis; neurological studies; behavior and cognition; reproduction; genetics; and xenotransplantation. Around 65,000 NHPs are used every year in the United States, and around 7,000 across the European Union. Most are purpose-bred, while some are caught in the wild.

Their use is controversial. According to the Nuffield Council on Bioethics, NHPs are used because their brains share structural and functional features with human brains, but "while this similarity has scientific advantages, it poses some difficult ethical problems, because of an increased likelihood that primates experience pain and suffering in ways that are similar to humans." Some of the most publicized attacks on animal research facilities by animal rights groups have occurred because of primate research. Some primate researchers have abandoned their studies because of threats or attacks.

In December 2006, an inquiry chaired by Sir David Weatherall, emeritus professor of medicine at Oxford University, concluded that there is a "strong scientific and moral case" for using primates in some research. The British Union for the Abolition of Vivisection argues that the Weatherall report failed to address "the welfare needs and moral case for subjecting these sensitive, intelligent creatures to a lifetime of suffering in UK labs".

Legal status

Further information: Great ape research ban

Human beings are recognized as persons and protected in law by the United Nations Universal Declaration of Human Rights and by all governments to varying degrees. Non-human primates are not classified as persons in most jurisdictions, which largely means their individual interests have no formal recognition or protection. The status of non-human primates has generated much debate, particularly through the Great Ape Project (GAP), which argues that great apes (gorillas, orangutans, chimpanzees, bonobos) should be given limited legal status and the protection of three basic interests: the right to live, the protection of individual liberty, and the prohibition of torture.

In 1997, the United Kingdom announced a policy of no longer granting licenses for research involving great apes, the first ever measure to ban primate use in research. Announcing the UK’s ban, the British Home Secretary said: "[T]his is a matter of morality. The cognitive and behavioural characteristics and qualities of these animals mean it is unethical to treat them as expendable for research." Britain continues to use other primates in laboratories, such as macaques and marmosets. In 2006 the permanency of the UK ban was questioned by Colin Blakemore, head of the Medical Research Council. Blakemore, while stressing he saw no "immediate need" to lift the ban, argued "that under certain circumstances, such as the emergence of a lethal pandemic virus that only affected the great apes, including man, then experiments on chimps, orang-utans and even gorillas may become necessary." The British Union for the Abolition of Vivisection described Blakemore's stance as "backward-looking." In 1999, New Zealand was the first country to ban experimentation on great apes by law.

On June 25, 2008, Spain became the first country to announce that it will extend rights to the great apes in accordance with GAP's proposals. An all-party parliamentary group advised the government to write legislation giving chimpanzees, gorillas and orangutans the right to life, to liberty, and the right not to be used in experiments. The New York Times reported that the legislation will make it illegal to kill apes, except in self-defense. Torture, which will include medical experiments, will be not allowed, as will arbitrary imprisonment, such as for circuses or films.

An increasing number of other governments are enacting bans. As of 2006, Austria, New Zealand (restrictions on great apes only and not a complete ban), the Netherlands, Sweden, and the United Kingdom had introduced either de jure or de facto bans. The ban in Sweden does not extend to non-invasive behavioral studies, and graduate work on great ape cognition in Sweden continues to be carried out on zoo gorillas, and supplemented by studies of chimpanzees held in the U.S. Sweden's legislation also bans invasive experiments on gibbons.

In December 2005, Austria outlawed experiments on any apes, unless it is conducted in the interests of the individual animal. In 2002, Belgium announced that it was working toward a ban on all primate use, and in the UK, 103 MPs signed an Early Day Motion calling for an end to primate experiments, arguing that they cause suffering and are unreliable. No licenses for research on great apes have been issued in the UK since 1998. The Boyd Group, a British group comprising animal researchers, philosophers, primatologists, and animal advocates, has recommended a global prohibition on the use of great apes.

The use of non-human primates in the EU is regulated under the Directive 2010/63/EU. The directive took effect on January 1, 2013. The directive permits the use of non-human primates if no other alternative methods are available. Testing on non-human primates is permitted for basic and applied research, quality and safety testing of drugs, food and other products and research aimed on the preservation of the species. The use of great apes is generally not permitted, unless it is believed that the actions are essential to preserve the species or in relation to an unexpected outbreak of a life-threatening or debilitating clinical condition in human beings. The directive stresses the use of the 3R principle (replacement, refinement, reduction) and animal welfare when conducting animal testing on non-human primates.

A 2013 amendment to the German Animal Welfare Act, with special regulations for monkeys, resulted in a near total ban on the use of great apes as laboratory animals. The last time great apes were used in laboratory experiments in Germany was 1991.

Species and numbers used

Covance primate-testing lab, Vienna, Virginia, 2004–05

Most of the NHPs used are one of three species of macaques, accounting for 79% of all primates used in research in the UK, and 63% of all federally funded research grants for projects using primates in the U.S. Lesser numbers of marmosets, tamarins, spider monkeys, owl monkeys, vervet monkeys, squirrel monkeys, and baboons are used in the UK and the U.S. Great apes have not been used in the UK since a government policy ban in 1998. In the U.S., research laboratories employ the use of 1,133 chimpanzees as of October 2006.

Data reported by region

Country	Total	Reporting Year	Procedures/Animals
Austria	0	2017	Procedures
Belgium	40	2016	Procedures
Bulgaria	0	2017	Procedures
Canada	7556	2016	Animals
Croatia	0	2016	Procedures
Cyprus	0	2016	Procedures
Czech Republic	36	2017	Animals
Denmark	0	2016	Procedures
Estonia	0	2016	Procedures
Finland	0	2016	Procedures
France	3508	2016	Procedures
Germany	2418	2016	Procedures
Greece	3	2016	Procedures
Hungary	0	2016	Procedures
Ireland	0	2016	Procedures
Israel	35	2017	Animals
Italy	511	2016	Procedures
Latvia	0	2016	Procedures
Lithuania	0	2015	Procedures
Luxembourg	0	2017	Animals
Malta	0	2016	Procedures
Netherlands	120	2016	Procedures
New Zealand	0	2015	Animals
Poland	0	2016	Procedures
Portugal	0	2014	Procedures
Romania	0	2015	Procedures
Slovakia	0	2017	Procedures
Slovenia	0	2016	Procedures
Spain	228	2016	Procedures
South Korea	2403	2017	Animals
Sweden	38	2016	Procedures
Switzerland	181	2017	Animals
United Kingdom	2960	2017	Procedures
United States	71188	2016	Animals

Most primates are purpose-bred, while some are caught in the wild. In 2011 in the EU, 0.05% of animals used in animal testing procedures were non-human primates.

In 1996, the British Animal Procedures Committee recommended new measures for dealing with NHPs. The use of wild-caught primates was banned, except where "exceptional and specific justification can be established"; specific justification must be made for the use of Old World primates (but not for the use of New World primates); approval for the acquisition of primates from overseas is conditional upon their breeding or supply center being acceptable to the Home Office; and each batch of primates acquired from overseas must be separately authorized.

Prevalence

There are indications that NHP use is on the rise in some countries, in part because biomedical research funds in the U.S. have more than doubled since the 1990s. In 2000, the NIH published a report recommending that the Regional Primate Research Center System be renamed the National Primate Research Center System and calling for an increase in the number of NHPs available to researchers, and stated that "nonhuman primates are crucial for certain types of biomedical and behavioral research." This assertion has been challenged. In the U.S., the Oregon and California National Primate Research Centers and New Iberia Research Center have expanded their facilities.

In 2000 the National Institutes of Health (NIH) invited applications for the establishment of new breeding specific pathogen free colonies; and a new breeding colony projected to house 3,000 NHPs has been set up in Florida. The NIH's National Center for Research Resources claimed a need to increase the number of breeding colonies in its 2004–2008 strategic plan, as well as to set up a database, using information provided through a network of National Primate Research Centers, to allow researchers to locate NHPs with particular characteristics. China is also increasing its NHP use, and is regarded as attractive to Western companies because of the low cost of research, the relatively lax regulations and the increase in animal-rights activism in the West.

In 2013, British Home Office figures show that the number of primates used in the UK was at 2,440, down 32% from 3,604 NHPs in 1993. Over the same time period, the number of procedures involving NHPs fell 29% from 4,994 from to 3,569 procedures.

Sources

Further information: Laboratory animal sources, Nafovanny, Shamrock Farm, and International primate trade

The American Society of Primatologists writes that most NHPs in laboratories in the United States are bred domestically. Between 12,000–15,000 are imported each year, specifically rhesus macaque monkeys, cynomolgus (crab-eating) macaque monkeys, squirrel monkeys, owl monkeys, and baboons. Monkeys are imported from China, Mauritius, the Philippines, and Peru.

China exported over 12,000 macaques for research in 2001 (4,500 to the U.S.), all from self-sustaining purpose-bred colonies. The second largest source is Mauritius, from which 3,440 purpose-bred cynomolgus macaques were exported to the U.S. in 2001.

In Europe, an estimated 70% of research primates are imported, and the rest are purpose-bred in Europe. Around 74% of these imports come from China, with most of the rest coming from Mauritius.

Use

General

NHPs are used in research into HIV, neurology, behavior, cognition, reproduction, Parkinson's disease, stroke, malaria, respiratory viruses, infectious disease, genetics, xenotransplantation, drug abuse, and also in vaccine and drug testing. According to The Humane Society of the United States, chimpanzees are most often used in hepatitis research, and monkeys in SIV research. Animals used in hepatitis and SIV studies are often caged alone.

Eighty-two percent of primate procedures in the UK in 2006 were in applied studies, which the Home Office defines as research conducted for the purpose of developing or testing commercial products. Toxicology testing is the largest use, which includes legislatively required testing of drugs. The second largest category of research using primates is "protection of man, animals, or environment", accounting for 8.9% of all procedures in 2006. The third largest category is "fundamental biological research", accounting for 4.9% of all UK primate procedures in 2006. This includes neuroscientific study of the visual system, cognition, and diseases such as Parkinson's, involving techniques such as inserting electrodes to record from or stimulate the brain, and temporary or permanent inactivation of areas of tissue.

Primates are the species most likely to be re-used in experiments. The Research Defence Society writes that re-use is allowed if the animals have been used in mild procedures with no lasting side-effects. This is contradicted by Dr. Gill Langley of the British Union for the Abolition of Vivisection, who gives as an example of re-use the licence granted to Cambridge University to conduct brain experiments on marmosets. The protocol sheet stated that the animals would receive "multiple interventions as part of the whole lesion/graft repair procedure." Under the protocol, a marmoset could be given acute brain lesions under general anaesthetic, followed by tissue implantation under a second general anaesthetic, followed again central cannula implantation under a third. The re-use is allowable when required to meet scientific goals, such as this case in which some procedures are required as preparatory for others.

Methods of restraint

A primate trained to place his head and hands in holes in the front of his cage. The holes are placed in such a way as to allow the primate to reach for food while presenting his head for the experiment.

One of the disadvantages of using NHPs is that they can be difficult to handle, and various methods of physical restraint have to be used. Viktor Reinhardt of the Wisconsin Regional Primate Research Center writes that scientists may be unaware of the way in which their research animals are handled, and therefore fail to take into account the effect the handling may have had on the animals' health, and thereby on any data collected. Reinhardt writes that primatologists have long recognized that restraint methods may introduce an "uncontrolled methodological variable", by producing resistance and fear in the animal. "Numerous reports have been published demonstrating that non-human primates can readily be trained to cooperate rather than resist during common handling procedures such as capture, venipuncture, injection and veterinary examination. Cooperative animals fail to show behavioural and physiological signs of distress."

Reinhardt lists common restraint methods as: squeeze-back cages, manual restraint, restraint boards, restraint chairs, restraint chutes, tethering, and nets. Alternatives include:

• chemical restraint; for example, ketamine, a sedative, may be given to the animal before a restraint procedure, reducing stress-hormone production;
• psychological support, in which an animal under restraint has visual and auditory contact with the animal's cage-mate. Blood pressure and heart rate responses to restraint have been measurably reduced using psychological support.
• training animals to cooperate with restraint. Such methods have been used and resulted in unmeasurable stress hormone responses to venipuncture, and no notable distress to being captured in a transport box.

Chimpanzees in the United States

Chimpanzee Enos, the third primate to orbit Earth, before the 1961 flight of Mercury-Atlas 5

As of 2013, the U.S. and Gabon were the only countries that still allowed chimpanzees to be used for medical experiments. The U.S. is the world's largest user of chimpanzees for biomedical research, with approximately 1,200 individual subjects in U.S. labs as of middle 2011, dropping to less than 700 as of 2016. Japan also still keeps a dozen chimpanzees in a research project for chimpanzee cognition (see Ai (chimpanzee)).

Chimpanzees routinely live 30 years in captivity, and can reach 60 years of age.

Most of the labs either conduct or make the chimpanzees available for invasive research, defined as "inoculation with an infectious agent, surgery or biopsy conducted for the sake of research and not for the sake of the chimpanzee, and/or drug testing." Two federally funded laboratories have used chimps: Yerkes National Primate Research Center at Emory University in Atlanta, Georgia, and the Southwest National Primate Research Center in San Antonio, Texas. By 2008, five hundred chimps had been retired from laboratory use in the U.S. and live in sanctuaries in the U.S. or Canada.

Their importation from the wild was banned in 1973. From then until 1996, chimpanzees in U.S. facilities were bred domestically. Some others were transferred from the entertainment industry to animal testing facilities as recently as 1983, although it is not known if any animals that were transferred from the entertainment industry are still in testing centers. Animal sanctuaries were not an option until the first North American sanctuary that would accept chimpanzees opened in 1976.

In 1986, to prepare for research on AIDS, the U.S. bred them aggressively, with 315 breeding chimpanzees used to produce 400 offspring. By 1996, it was clear that SIV/HIV-2/SHIV in macaque monkeys was a preferred scientific AIDS model to the chimpanzees, which meant there was a surplus. A five-year moratorium on breeding was therefore imposed by the U.S. National Institutes of Health (NIH) that year, and it has been extended annually since 2001. As of October 2006, the chimpanzee population in US laboratories had declined to 1133 from a peak of 1500 in 1996.

Chimpanzees tend to be used repeatedly over decades, rather than used and killed as with most laboratory animals. Some individual chimpanzees currently in U.S. laboratories have been used in experiments for over 40 years. The oldest known chimpanzee in a U.S. lab is Wenka, who was born in a laboratory in Florida on May 21, 1954. She was removed from her mother on the day of birth to be used in a vision experiment that lasted 17 months, then sold as a pet to a family in North Carolina. She was returned to the Yerkes National Primate Research Center in 1957 when she became too big to handle. Since then, she has given birth six times, and has been used in research into alcohol use, oral contraceptives, aging, and cognitive studies.

With the publication of the chimpanzee genome, there are reportedly plans to increase the use of chimpanzees in labs, with scientists arguing that the federal moratorium on breeding chimpanzees for research should be lifted. Other researchers argue that chimpanzees are unique animals and should either not be used in research, or should be treated differently. Pascal Gagneux, an evolutionary biologist and primate expert at the University of California, San Diego, argues that, given chimpanzees' sense of self, tool use, and genetic similarity to human beings, studies using chimpanzees should follow the ethical guidelines that are used for human subjects unable to give consent. Stuart Zola, director of the Yerkes National Primate Research Laboratory, disagrees. He told National Geographic: "I don't think we should make a distinction between our obligation to treat humanely any species, whether it's a rat or a monkey or a chimpanzee. No matter how much we may wish it, chimps are not human."

In January 2011 the Institute of Medicine was asked by the NIH to examine whether the government should keep supporting biomedical research on chimpanzees. The NIH called for the study after protests by the Humane Society of the United States, primatologist Jane Goodall and others, when it announced plans to move 186 semi-retired chimpanzees back into active research. On December 15, 2011, the Institute of Medicine committee concluded in their "Chimpanzees in Biomedical and Behavioral Research: Assessing the Necessity" report that, "while the chimpanzee has been a valuable animal model in past research, most current use of chimpanzees for biomedical research is unnecessary," as scientific research indicated a decreasing need for the use of chimpanzees due to the emergence of non-chimpanzee models. Later that day Francis Collins, a head of the NIH, said the agency would stop issuing new awards for research involving chimpanzees until the recommendations developed by the IOM are implemented.

On 21 September 2012, the NIH announced that 110 chimpanzees owned by the government were to be retired. The NIH owned about 500 chimpanzees for research, and this move signified the first step to wind down its investment in chimpanzee research, according to Collins. Housed at the New Iberia Research Center in Louisiana, 10 of the retired chimpanzees were to go to the chimpanzee sanctuary Chimp Haven while the rest were to go to Texas Biomedical Research Institute in San Antonio. However, concerns over the chimpanzees' status in the Texas Biomedical Research Institute as ‘research ineligible’ rather than ‘retired’ prompted the NIH to reconsider the plan. On 17 October 2012, it was announced that as many chimpanzees as possible will be relocated to Chimp Haven by August 2013, and that eventually all 110 will move there.

In 2013 the NIH agreed with the IOM's recommendations that experimentation on chimpanzees was unnecessary and rarely helped in advancing human health for infectious diseases and that the NIH would phase out most of its government-funded experiments on chimpanzees. On 22 January 2013, an NIH task force released a report calling for the government to retire most of the chimpanzees under U.S. government support. The panel concluded that the animals provide little benefit in biomedical discoveries except in a few disease cases which can be supported by a small population of 50 primates for future research. It suggested that other approaches, such as genetically altered mice, should be developed and refined instead. On 13 November 2013, Congress and the Senate passed ‘The Chimpanzee Health Improvement, Maintenance and Protection Act’, which approved funding to expand the capacity of Chimp Haven and other chimpanzee sanctuaries, allowing for the transfer of almost all of the apes owned by the federal government to live in a more natural and group environment. The transfer was expected to take up to five years, at which point all but 50 chimpanzees were to have been successfully ‘retired’.

On 11 June 2013, the US Fish and Wildlife Service (USFWS) proposed to list captive chimpanzees as endangered, matching its existing classification for wild chimpanzees. Until the USFWS proposal, chimpanzees were the only species with a split listing that did not also classify captive members of the species as endangered. Before the proposal gained final approval, it was unclear what effect it would have on laboratory research.

Two years later, on June 16, 2015, the USFWS announced that it has designated both captive and wild chimpanzees as endangered. In November 2015 the NIH announced it would no longer support biomedical research on chimpanzees and release its remaining 50 chimpanzees to sanctuaries. The agency would also develop a plan for phasing out NIH support for the remaining chimps that are supported by, but not owned by, the NIH.

In January 2014, Merck & Co. announced that the company will not use chimpanzees for research, joining over 20 pharmaceutical companies and contract laboratories that have made the commitment. As the trend continues, it is estimated the remaining non-government owned 1,000 chimpanzees will be retired to sanctuaries around 2020.

Notable studies

Polio

In the 1940s, Jonas Salk used rhesus monkey cross-contamination studies to isolate the three forms of the polio virus that crippled hundreds of thousands of people yearly across the world at the time. Salk's team created a vaccine against the strains of polio in cell cultures of green monkey kidney cells. The vaccine was made publicly available in 1955, and reduced the incidence of polio 15-fold in the USA over the following five years.

Albert Sabin made a superior "live" vaccine by passing the polio virus through animal hosts, including monkeys. The vaccine was produced for mass consumption in 1963 and is still in use today. It had virtually eradicated polio in the United States by 1965.

Split-brain experiments

In the 1950s, Roger Sperry developed split-brain preparations in non-human primates that emphasized the importance of information transfer that occurred in these neocortical connections. For example, learning on simple tasks, if restricted in sensory input and motor output to one hemisphere of a split-brain animal, would not transfer to the other hemisphere. The right brain has no idea what the left brain is up to, if these specific connections are cut. Those experiments were followed by tests on human beings with epilepsy who had undergone split-brain surgery, which established that the neocortical connections between hemispheres are the principal route for cognition to transfer from one side of the brain to another. These experiments also formed the modern basis for lateralization of function in the human brain.

Vision experiments

In the 1960s, David Hubel and Torsten Wiesel demonstrated the macrocolumnar organization of visual areas in cats and monkeys, and provided physiological evidence for the critical period for the development of disparity sensitivity in vision (i.e., the main cue for depth perception). They were awarded a Nobel Prize for their work.

Deep-brain stimulation

In 1983, designer drug users took MPTP, which created a Parkinsonian syndrome. Later that same year, researchers reproduced the effect in non-human primates. Over the next seven years, the brain areas that were over- and under-active in Parkinson's were mapped out in normal and MPTP-treated macaque monkeys using metabolic labelling and microelectrode studies. In 1990, deep brain lesions were shown to treat Parkinsonian symptoms in macaque monkeys treated with MPTP, and these were followed by pallidotomy operations in humans with similar efficacy. By 1993, it was shown that deep brain stimulation could effect the same treatment without causing a permanent lesion of the same magnitude. Deep brain stimulation has largely replaced pallidotomy for treatment of Parkinson's patients that require neurosurgical intervention. Current estimates are that 20,000 Parkinson's patients have received this treatment.

AIDS

The non-human primate models of AIDS, using HIV-2, SHIV, and SIV in macaques, have been used as a complement to ongoing research efforts against the virus. The drug tenofovir has had its efficacy and toxicology evaluated in macaques, and found longterm-highdose treatments had adverse effects not found using short term-high dose treatment followed by long term-low dose treatment. This finding in macaques was translated into human dosing regimens. Prophylactic treatment with anti-virals has been evaluated in macaques, because introduction of the virus can only be controlled in an animal model. The finding that prophylaxis can be effective at blocking infection has altered the treatment for occupational exposures, such as needle exposures. Such exposures are now followed rapidly with anti-HIV drugs, and this practice has resulted in measurable transient virus infection similar to the NHP model. Similarly, the mother-to-fetus transmission, and its fetal prophylaxis with antivirals such as tenofovir and AZT, has been evaluated in controlled testing in macaques not possible in humans, and this knowledge has guided antiviral treatment in pregnant mothers with HIV. "The comparison and correlation of results obtained in monkey and human studies is leading to a growing validation and recognition of the relevance of the animal model. Although each animal model has its limitations, carefully designed drug studies in nonhuman primates can continue to advance our scientific knowledge and guide future clinical trials."

Treatment of anxiety and depression

The reason for studying primates is due to the similar complexity of the cerebral processes in the human brain which controls emotional responses and can be beneficial for testing new pharmacological treatments. An experiment published in the Neuroscience & Biobehavioral Reviews describes habituation of the black-tufted marmoset in a figure eight maze model. They were presented with a taxidermized wild-cat, rattlesnake, a hawk as well as a stuffed toy bear on one side of the maze. Two cameras and a two way mirror was used to observe the difference between the monkeys natural behaviors versus the behaviors expressed by the diazepam induced monkeys in thirteen different locations inside the maze. Scientist Barros and his colleagues created this model to allow the monkeys to roam a less confined environment and slightly eliminate outside factors that may induce stress.

Allegations

Many of the best-known allegations of abuse made by animal protection or animal rights groups against animal-testing facilities involve non-human primates.

University of Wisconsin–Madison

Main article: Pit of despair

The so-called "pit of despair" was used in experiments conducted on rhesus macaque monkeys during the 1970s by American comparative psychologist Harry Harlow at the University of Wisconsin–Madison. The aim of the research was to produce clinical depression. The vertical chamber was a stainless-steel bin with slippery sides that sloped to a rounded bottom. A 3/8 in. wire mesh floor 1 in. above the bottom of the chamber allowed waste material to drop out of holes. The chamber had a food box and a water-bottle holder, and was covered with a pyramid top so that the monkeys were unable to escape. Harlow placed baby monkeys in the chamber alone for up to six weeks. Within a few days, they stopped moving about and remained huddled in a corner. The monkeys generally exhibited marked social impairment and peer hostility when removed from the chamber; most did not recover.

University of California, Riverside

Main article: University of California Riverside 1985 laboratory raid

On April 21, 1985, activists of the Animal Liberation Front (ALF) broke into the UC Riverside laboratories and removed hundreds of animals. According to Vicky Miller of PETA, who reported the raid to newswire services, UC-Riverside "has been using animals in experiments on sight deprivation and isolation for the last two years and has recently received a grant, paid for with our tax dollars, to continue torturing and killing animals." According to UCR officials, the ALF claims of animal mistreatment were "absolutely false," and the raid would result in long-term damage to some of the research projects, including those aimed at developing devices and treatment for blindness. UCR officials also reported the raid also included smashing equipment and resulted in several hundred thousand dollars of damage.

Covance

Main article: Covance

In Germany in 2004, journalist Friedrich Mülln took undercover footage of staff in Covance in Münster, Europe's largest primate-testing center. Staff were filmed handling monkeys roughly, screaming at them, and making them dance to blaring music. The monkeys were shown isolated in small wire cages with little or no natural light, no environmental enrichment, and subjected to high noise levels from staff shouting and playing the radio. Primatologist Jane Goodall described their living conditions as "horrendous."

A veterinary toxicologist employed as a study director at Covance in Vienna, Virginia, from 2002 to 2004, told city officials in Chandler, Arizona, that Covance was dissecting monkeys while the animals were still alive and able to feel pain. The employee approached the city with her concerns when she learned that Covance planned to build a new laboratory in Chandler.

She alleged that three monkeys in the Vienna laboratory had pushed themselves up on their elbows and had gasped for breath after their eyes had been removed, and while their intestines were being removed during necropsies (autopsy). When she expressed concern at the next study directors' meeting, she says she was told that it was just a reflex. She told city officials that she believed such movements were not reflexes but suggested "botched euthanasia performed by inadequately trained personnel." She alleged that she was ridiculed and subjected to thinly veiled threats when she contacted her supervisors about the issue.

University of Cambridge

BUAV alleges that monkeys were left unattended for up to 15 hours after having parts of their brains removed to induce strokes.

 

Main article: Primate experiments at Cambridge University

In the UK, after an undercover investigation in 1998, the British Union for the Abolition of Vivisection (BUAV), a lobby group, reported that researchers in Cambridge University's primate-testing labs were sawing the tops off marmosets' heads, inducing strokes, then leaving them overnight without veterinarian care, because staff worked only nine to five. The experiments used marmosets that were first trained to perform certain behavioral and cognitive tasks, then re-tested after brain damage to determine how the damage had affected their skills. The monkeys were deprived of food and water to encourage them to perform the tasks, with water being withheld for 22 out of every 24 hours.

The Research Defence Society defended Cambridge's research. The RDS wrote that the monkeys were fully anaesthetised, and appropriate pain killers were given after the surgery. "On recovery from the anaesthesia, the monkeys were kept in an incubator, offered food and water and monitored at regular intervals until the early evening. They were then allowed to sleep in the incubators until the next morning. No monkeys died unattended during the night after stroke surgery." A court rejected BUAV's application for a judicial review. BUAV appealed.

Columbia University

Main article: Primate experiments at Columbia University

In 2003, CNN reported that a post-doctoral veterinarian at Columbia University complained to the university's Institutional Animal Care and Use Committee about experiments being conducted on baboons by E. Sander Connolly, an assistant professor of neurosurgery. The experiment involved a left transorbital craniectomy to expose the left internal carotid artery to occlude the blood supply to the brain. A clamp was placed on this blood vessel until the stroke was induced, after which Connolly would test a potential neuroprotective drug which if effective, would be used to treat humans suffering from stroke.

Connolly developed this methodology to make more consistent stroke infarcts in primates, which would improve the detection of differences in stroke treatment groups, and "provide important information not obtainable in rodent models." The baboons were kept alive after the surgery for observation for three to ten days in a state of "profound disability" which would have been "terrifying," according to neurologist Robert Hoffman. Connolly's published animal model states that animals were kept alive for three days, and that animals that were successfully self-caring were kept alive for 10 days. People for the Ethical Treatment of Animals has expressed strong opposition to this experiment and has written multiple letters to the NIH and other federal agencies to halt further mistreatment of baboons and other animals at Columbia.

An investigation by the U.S. Department of Agriculture found "no indication that the experiments...violated federal guidelines." The Dean of Research at Columbia's School of Medicine said that Connolly had stopped the experiments because of threats from animal rights activists, but still believed his work was humane and potentially valuable.

Attacks on researchers

In 2006, activists forced a primate researcher at UCLA to shut down the experiments in his lab. His name, phone number, and address were posted on the website of the UCLA Primate Freedom Project, along with a description of his research, which stated that he had "received a grant to kill 30 macaque monkeys for vision experiments. Each monkey is first paralyzed, then used for a single session that lasts up to 120 hours, and finally killed." Demonstrations were held outside his home. A Molotov cocktail was placed on the porch of what was believed to be the home of another UCLA primate researcher. Instead, it was accidentally left on the porch of an elderly woman unrelated to the university. The Animal Liberation Front claimed responsibility for the attack.

As a result of the campaign, the researcher sent an email to the Primate Freedom Project stating "you win", and "please don't bother my family anymore." In another incident at UCLA in June 2007, the Animal Liberation Brigade placed a bomb under the car of a UCLA children's ophthalmologist, who performs experiments on cats and rhesus monkeys; the bomb had a faulty fuse and did not detonate. UCLA is now refusing Freedom of Information Act requests for animal medical records.

The house of UCLA researcher Edythe London was intentionally flooded on October 20, 2007, in an attack claimed by the Animal Liberation Front. London conducts research on addiction using non-human primates, and no claims were made by the ALF of any violation of any rules or regulations regarding the use of animals in research. London responded by writing an op-ed column in the LA Times titled "Why I use laboratory animals."

In 2009, a UCLA neurobiologist known for using animals to research drug addiction and other psychiatric disorders had his car burned for the second time.

China

In infectious disease research, China invests more than the U.S. does in conducting research on non-human primates. "Select agents and toxins" refers to a list of over 60 substances that pose the greatest risk to public health, and China uses non-human primates to test treatment of these select agents and toxins more than the U.S. does.