bluuebunny/crossref_metadata_2025 huggingface dataset:
An In Vitro Phototoxicity Assay Battery (Photohaemolysis and 3T3 NRU PT test) to Assess Phototoxic Potential of Fragrances
Authors: Nam Chunja , An Susun , Lee Eunyoung , Moon Seongjoon , Kang Jongkoo , Chang Ihseop
Published: April 2019

The purpose of this study was to compare the in vivo and in vitro phototoxicity potentials of 13 fragrances. We used the 3T3 neutral red uptake phototoxicity (3T3 NRU PT) test and the photohaemolysis test as in vitro phototoxicity assays. In the 3T3 NRU PT test, all of the fragrances were non-phototoxic. Six fragrances were phototoxic in the photohaemolysis test. Three of the six photohaemolytic fragrances were phototoxic in the guinea-pig photoirritation test. These phototoxic fragrances did not cause cellular phototoxicity, but showed a photohaemolytic reaction. The photohaemolysis test was more sensitive than the 3T3 NRU PT test for screening for the phototoxicity of fragrances. The accuracy of this in vitro phototoxicity test battery was 82%. It is thought that the major phototoxic mechanism of fragrances is cell membrane damage. We suggest that a battery composed of the 3T3 NRU PT test and the photohaemolysis test is a simple and effective model for the in vitro phototoxicity assay of fragrances.
Summary of the Workshop on Training for the Implementation of Alternative Test Methods
Authors: Halder Marlies , Sailstad Denise
Published: April 2019

Over the last two decades, emphasis has been on new method validation, acceptance criteria and international harmonisation, and in recent years, several scientifically validated alternative test methods have been accepted. The objectives of this workshop were to review the various training programmes related to reduction, refinement and replacement (the Three Rs) and validated alternative methods, the obstacles to such training and ways to overcome them. In general, training in the Three Rs should be a continuous exercise. Whenever the regulators accept an alternative test method, specific training should be offered in order to overcome the reluctance to change and to expedite the implementation of the new test method. It is essential that funds be made available for this purpose, e.g. via regulatory agencies and international organisations.
Report of the Workshop on the Validation of QSARs and Other Computational Prediction Models
Authors: Worth Andrew P. , Cronin Mark T.D.
Published: April 2019

This workshop addressed current issues with regard to establishing a framework for the validation of quantitative structure–activity relationships (QSARs) and other computational prediction models. QSARs and related models attempt to associate physicochemical and structural properties of compounds to their biological activity. As such, they may permit the prediction of biological activity from chemical structure alone. As yet, no formal validation criteria have been agreed internationally for QSARs and related techniques. However, some general and preliminary criteria were discussed and agreed upon at a European Chemical Industry Council/International Council of Chemical Associations (CEFIC/ICCA) workshop on the regulatory acceptance of QSARs, held in Setubal, Portugal, in March 2002. These criteria were presented at a Fourth World Congress workshop, along with a proposal for the practical validation of computer models, such as QSARs.
The ICLAS/CCAC International Symposium on Regulatory Testing and Animal Welfare
Authors: Griffin Gilly , Stokes William S. , Pakes Steven P. , Gauthier Clément
Published: April 2019

The first International Symposium on Regulatory Testing and Animal Welfare (ISRTAW), held 21–23 June 2001, in Québec City, Canada, brought together 160 experts from 22 countries from North and South America, Europe and Asia. The experts included representatives from national research and regulatory agencies, universities, and industry involved in chemicals, pesticides and drug safety testing. Representatives from European, Canadian and US animal welfare groups also participated in the discussions. The Symposium was organised by the International Council for Laboratory Animal Science (ICLAS) and the Canadian Council on Animal Care (CCAC), with the support and assistance of many sponsors and advisors. ICLAS is a worldwide organisation whose purpose is to foster the international harmonisation of animal care and use practices. CCAC is the national agency responsible for overseeing the ethical use of animals in Canadian science. Both organisations are committed to fostering an environment in which global efforts to harmonise testing procedures using animals in a more-humane manner can be realised.
Full Thickness EpiDerm™: A Dermal–Epidermal Skin Model to Study Epithelial–Mesenchymal Interactions
Authors: Kubilus Joseph , Hayden Patrick J. , Ayehunie Seyoum , Lamore Sarah D.K. , Servattalab Cynthia , Bellavance Kristen L. , Sheasgreen John E. , Klausner Mitchell
Published: April 2019

Unlike previous dermal–epidermal models, Full Thickness EpiDerm™ is cultured in an easily manipulated cell culture insert, and the tissue extends from wall to wall. In terms of ease of use, these characteristics greatly facilitate the testing of potential allergens or irritants in that direct topical application is possible. Topical exposure to the common surfactant, 1% Triton X-100, resulted in MTT tissue viability dose–response curves that fell within the normal range of the keratinocyte-only tissue, EpiDerm. Currently, in order to produce a standardised, reproducible organotypic tissue, all lots of EpiDerm are compared to a reference database of effective time-50 (ET-50) values, i.e. the time of exposure after which viability is reduced to 50% following exposure to 100μl of Triton X-100. The database average (184 tissue lots) is 6.74 ± 0.99 hours (± 1 SD); initial lots of the full thickness tissue, tested in an identical manner, averaged 7.79 ± 1.24 hours (n = 11). Histological cross-sections of the full thickness tissue showed an epidermal layer that is very similar to EpiDerm and native epidermis on a fibroblast-containing collagen matrix dermis-like layer. Irradiation of the tissue with ultraviolet light induced an increase in collagenase (MMP-1) release. Based on these initial results, investigation of the tissue response to stimuli specifically affecting the dermis or epidermal/dermal “cross-talk” will probably prove informative.
Brueghel's Two Monkeys
Authors: Newkirk Ingrid E.
Published: April 2019

Progress in finding alternatives to animal tests continues to be painfully slow, and pain is certainly the operative word for animals involved in the tests. US government agencies are not simply being slow, they are not moving forward at all. For example, the US Environmental Protection Agency has been directed by Congress to spend a small amount of its budget on non-animal test method development, yet it has not done so, and the in vivo percutaneous absorption test continues to be used, despite the existence of an approved non-animal method. A movement is building that may compel industry to stop hiding behind the regulators and take action. There are excellent reasons, to stop fighting such advances. For ethical, practical, business and other reasons, it is time to enthusiastically embrace an effective approach to finding, advocating and adopting non-animal test methods.
The Use of Reconstructed Human Epidermis EPISKIN™ in the Assessment of Local Tolerance of Cosmetics and Chemicals
Authors: Roguet Roland
Published: April 2019

Models of reconstructed human skin provide useful tools for both basic research and evaluation of the safety and efficacy of dermato-cosmetological products. They can provide alternative models to replace human and animal experiments. However, their use requires confidence in their reproducibility and similarity of the results obtained as compared to in vivo experiments. Their morphology, architecture and properties must be as close as possible to human skin. The present paper reviews the results obtained on the EPISKIN™ model through characterisation of the model, reproducibility studies and multicentre studies or (pre)validations.
ECVAM's Activities in Promoting the Three Rs in the Quality Control of Biologicals
Authors: Halder Marlies , Balls Michael , Hendriksen Coenraad , Cussler Klaus
Published: April 2019

This paper summarises key activities initiated and the progress achieved between April 1993 and November 2002, in promoting the Three Rs in one of ECVAM's priority areas — the production and quality control of biologicals. These have included organising nine key workshops, financially supporting and/or participating in a number of prevalidation and/or validation studies, financial contributions and sponsorship to relevant international workshops, symposia and conferences, and financial support to the compilation of manuals, expert reports and training in test methods.
Rat Hepatocyte Suspensions as a Suitable In Vitro Model for Studying the Biotransformation of Histone Deacetylase Inhibitors
Authors: Elaut Greetje , Török Gabriëlla , Papeleu Peggy , Vanhaecke Tamara , Laus Georges , Tourwé Dirk , Rogiers Vera
Published: April 2019

This paper focuses on the use of liver-derived in vitro systems for biotransformation studies during early drug development, as exemplified by the two molecules recently studied in our laboratory: Trichostatin A (TSA) and its structural analogue 5-(4-dimethylaminobenzoyl)aminovaleric acid hydroxamide (4-Me2N-BAVAH). Phase I biotransformation of TSA, a histone deacetylase inhibitor with promising antifibrotic and antitumoural properties, was investigated in liver microsomal (rat and human) and in hepatocyte (rat) suspensions. Within 40 minutes, 50μM of TSA was completely metabolised by 2 × 106 hepatocytes/ml. Reduction of the hydroxamic acid function to its corresponding amide and N-demethylation were the two major phase I biotransformation pathways, while hydrolysis products of TSA were minor metabolites. Lower concentrations of TSA (5μM and 25μM) were N-demethylated faster. Liver microsomes, however, metabolised TSA incompletely with the formation of two major metabolites, N-mono- and N-didemethylated TSA. Unlike TSA, 4-Me2N-BAVAH (50μM) could still be detected after 3 hours of incubation with 2 × 106 rat hepatocytes/ml suspension. Hydrolysis and reduction of the hydroxamic acid function to its corresponding acid and amide, respectively, were shown to be the major phase I biotransformation pathways. Lower concentrations of 4-Me2N-BAVAH were hydrolysed more readily. 4-Me2N-BAVAH and its metabolites were less subjected to N-demethylation than TSA.
Methods for Evaluation of Laboratory Animal Well-being
Authors: Baumans Vera
Published: April 2019

Well-being is a relative concept, referring to the state of an animal in relation to its ability to cope with its environment. This ability to cope is what we usually try to measure when evaluating the animal's well-being. Good welfare is, in general, considered to be related to a broad behavioural repertoire, which requires a considerable knowledge of the animal's species-specific behaviour and their basic biology. Ideally, well-being should be measured in a positive way, such as measuring pleasure by anticipatory behaviour. However, parameters have more often been designed for detecting failures to cope, leading to stress and/or discomfort. Parameters used in the assessment of discomfort are behavioural parameters, such as stereotypies, reduction in grooming, changes in activity; physiological parameters, such as body weight, abnormal posture, respiratory signs, heart rate, hormone levels; and post-mortem signs, as retrospective parameters, such as stomach ulcers, adrenal cortex size, fatty deposits. The usefulness of these parameters is discussed.
Refinement of Gerbil Housing and Husbandry in the Laboratory
Authors: Waiblinger Eva , König Barbara
Published: April 2019

Mongolian gerbils, Meriones unguiculatus, are widely used as model species in parasitology, and in research on epilepsy and ischaemia. When kept under standard laboratory housing conditions, gerbils develop two distinctive behavioural abnormalities: stereotypic digging and bar chewing. Our work centred on assessing the mechanisms leading to these stereotypies. We aimed at finding practical housing and husbandry solutions that would prevent the development of such stereotypies in the laboratory by addressing the behavioural needs of gerbils. Faecal cortisol levels were used as a non-invasive method to compare stress reactions in gerbils housed under different conditions. Stereotypic digging was caused by the lack of a burrow in the laboratory cage, while stereotypic bar chewing was caused by premature separation of juveniles from their family before the birth of younger siblings. An integrated artificial burrow system is presented that prevents the development of stereotypic digging in gerbils, and adaptations in husbandry routines are suggested that significantly reduce the incidence of bar chewing. We conclude that simple refinement of housing is effective in improving gerbil welfare in the laboratory.
Use and Pitfalls of Allometry: A Valuable Tool in Comparisons and Extrapolations between Species and in Ethical Considerations concerning the use of One Species to Model Another
Authors: Kirkwood James K.
Published: April 2019

Where research on one species is justified on the grounds that it will provide benefits to another, the strength of the ethical case depends critically on whether findings can be extrapolated meaningfully. Valid extrapolation depends on the species being sufficiently similar in respects critical to the research and on knowledge of the bases and effects of salient differences. Many biological parameters vary with body weight ( W) between species. When species of small size are used to model larger ones, the influence of size on the rates of physiological, immunological and other processes must be taken into account. Between species, the rates of physiological processes tend to increase with W0.75, and the durations of physiological events tend to increase with W0.25. Providing potential pitfalls are understood, allometric scaling enables valid comparisons and extrapolations between species. Knowledge of these principles is crucial also in making predictions about many aspects of animals’ biology and has application also in making sound ethical judgments about the justifiability of extrapolations between species concerning the wide range of processes linked to rates of metabolism.
Welfare Benefits to Farm Animals from Their Use in Research
Authors: Colditz Ian G.
Published: April 2019

Domestication is the purposeful selection and modification of livestock to suit the human-made production environments. While the application of modern breeding theory has accelerated genetic change, traditional practices also modify the genetic makeup of livestock. Threats to the welfare of farm animals can arise from husbandry practices, attributes of the production environment and failure to match the genotype to the production environment. The selection of sheep for resistance to gastrointestinal nematodes, the yard weaning of calves and the selection of cattle with docile temperament (e.g. slow flight time) are described as methods for improving welfare outcomes for grazing sheep and feedlot cattle, respectively. Contemporary production breeds exist as global populations. The move to the life-long individual identification of animals improves the power of breeding and management programmes that satisfy the criteria of scientific experimentation in their manner of implementation. In contrast to the use of farm animals, prejudice exists against comparable uses of more-recent domesticates (e.g. laboratory rodents). The growing global demand for livestock products suggests that Russell & Burch's Three Rs no longer provide a valid framework for deliberation on ethics of domestic animal use. A welfare outcome metric, such as that adopted by Grandin, is proposed as more robust than input quantification (reduce, replace) for appraisal of the ethics of animal use.
Optimising Resources by Reduction: The FRAME Reduction Committee
Authors: Vaughan Sylvia
Published: April 2019

The Fund for the Replacement of Animals in Medical Experiments (FRAME) Reduction Committee, formed in 1998, comprises experienced professionals in the fields of statistics, experimental design, animal welfare and alternatives. The committee undertakes projects designed to contribute to reduction by addressing the implementation of recommendations made at an European Centre for the Validation of Alternative Methods (ECVAM) workshop on reducing the use of laboratory animals. The use of improved experimental design and statistical analysis techniques are key means of achieving reduction. Literature surveys have shown that there is scope for improvement in these areas. Projects include organising workshops that explain how these techniques can be used to achieve reduction, the creation of guidelines for journal editors and the compilation of a directory of training material on experimental design and statistical analysis. The first FRAME Reduction Committee international symposium will be held in October 2002. 1
Refinement and Reduction through the Control of Variation
Authors: Festing Michael F.W.
Published: April 2019

The key to doing animal experiments efficiently, while using the minimum number of animals without loss of scientific information, lies in good control of random variation, and recognition and control of “fixed effect” variation, such as the sex or strain of the animals. However, many scientists erroneously assume that the use of outbred, genetically heterogeneous animals is justified, because in some way, they more closely model humans. Unfortunately, all this does is to increase the phenotypic variation, which results in less-powerful experiments. If the aim is to model variation in human responses, this can be done by using a small number of animals from several isogenic strains, without increasing the total number of animals. Reducing inter-individual variation, whether caused by genetic or non-genetic causes, will nearly always result in improved experiments. Fixed-effect variation, such as the sex of the animals, can be taken into account, either by restricting the conclusions to the sex actually used, or by assuming that the other sex would respond in the same way, or by including both sexes in the study, by using a factorial design, without increasing the total number of animals.
Physiological and Behavioural Assessment of Pain in Ruminants: Principles and Caveats
Authors: Mellor David J. , Stafford Kevin J.
Published: April 2019

Pain elicits a range of physiological and behavioural responses. These are commonly used to assess the impact of pain-inducing stimuli on animals, to determine whether or not significant pain is experienced and to devise strategies for alleviating pain. This paper outlines a range of principles and caveats to guide the evaluation of physiological and behavioural responses to painful stimuli, so that they can be better used to minimise pain in the experimental context. Although this advice is based on studies of farm animals responding to painful husbandry practices, it is more generally applicable.
The Welfare of Non-human Primates in Research in the EU
Authors: Morton David B.
Published: April 2019

This summarises a draft report of the Non-human Primate Working Group, prepared for the European Union's Scientific Committee on Animal Health and Welfare. It was formally adopted by the Scientific Committee at their meeting in December 2002, and the full text is available online at http://europa.eu.int/comm/food/fs/sc/scah/out83_en.pdf .
The Moral Standing of Non-human Primates: Why They Merit Special Consideration
Authors: Sauer Ursula G.
Published: April 2019

Scientific experiments with non-human primates are viewed very controversially. Those who use non-human primates for scientific purposes contend that the results will be of great benefit to humans. They say that the distress to the animals is minimal and that, therefore, their experiments are ethically acceptable. On the other hand, there are those who oppose non-human primate experiments altogether, due to the close relationship between these animals and humans. In a literature study on non-human primate experiments in fundamental brain research, the outcome of research projects performed over the period of a decade was evaluated retrospectively. Objective criteria were applied both for determining the cost (internationally published distress scores, taking into account distress due to procurement, transportation and housing) and the benefit (evaluating publications that arose from the experiments, citation scores and so forth). Taking the example of electrophysiological recordings in the brain of the awake macaque, this balancing ended up very different from that proposed by the scientists, and did not support the conclusion that the experiments had been ethically acceptable.
Animal Use in Biotechnology: Issues and Options — A New Zealand Perspective
Authors: Bayvel A.C. David
Published: April 2019

Although New Zealand has a relatively small national biotechnology research effort, there are a number of specific areas that have attracted attention in recent years. This has resulted from the interests and expertise of individual researchers, the priorities of both public and private sector research institutions, the importance of the livestock industries to the New Zealand economy, the high health status of New Zealand's cattle and sheep populations and, particularly, the absence of scrapie and bovine spongi-form encephalopathy. A number of research activities are reviewed, with particular reference to the associated welfare and ethical issues. This includes reference to relevant legislation covering animal welfare and new organisms. Particular attention is given to public and political interest in the newer research areas. The challenges posed to researchers and regulators in relation to both public consultation and effective communication of the risks and benefits of the work proposed or being undertaken are addressed. The role of independent expert advisory committees (the National Animal Ethics Advisory Committee and the National Animal Welfare Advisory Committee) and the need for lay members of animal ethics committees to be fully conversant with the welfare implications and ethical considerations of the technology being used are also discussed.
Ethics and Transgenesis: Toward a Policy Framework Incorporating Intrinsic Objections and Societal Perceptions
Authors: Gott Miranda , Monamy Vaughan
Published: April 2019

Concerns about the use of transgenic animals can be broadly separated into extrinsic factors, such as risks to human and non-human health, environment, economy and society, and intrinsic factors, such as spiritual, cultural, moral or otherwise personal beliefs about humans, animals and Nature. We examine the bases for making ethical decisions about the use of transgenic animals and the intrinsic beliefs that underpin common objections to their use. We explore the need for a policy framework based on a utilitarian approach to ethical decision-making, which balances costs with benefits, but which also acknowledges the intrinsic beliefs that lie at the core of much opposition to genetic modification of animals. In particular circumstances, intrinsic objections may lead to a society refusing to condone transgenesis regardless of the weight of benefit promised from proposed uses. In sensitive subject areas such as regulating the use of transgenic animals, public policy is hard-pressed to keep up with the rapid pace of technological change. We advocate the use of dynamic processes that are capable of reflecting shifting public attitudes.
Report of the Workshop on Euthanasia Guidelines and Practices
Authors: Carbone Larry , Baumans Vera , Morton David B.
Published: April 2019

Determining ethical standards for laboratory animal euthanasia requires an assessment of the relative amounts of pain and distress caused by different methods. Animal behaviour data are an important indicator of pain and distress, but their interpretation can be controversial; moreover, behaviour is more easily assessed with some euthanasia methods than with others. While every euthanasia method requires careful study, CO2 inhalation has come under close scrutiny both because it is so widely used for rodent euthanasia, and because it has, until recently, long been considered relatively non-aversive.
Comprehensive Assessment of Harms Caused by Experimental, Teaching and Testing Procedures on Live Animals
Authors: Mellor David J.
Published: April 2019

The comprehensiveness of assessments of experimental, teaching and testing harms, and the actions taken to minimise them, are measures of the scientist's acceptance of ethical responsibility for all features of each procedure that affect animals adversely. A system, refined from its original 1994 form, has been devised to assist in this process. Five “domains of potential animal welfare compromise” are identified. Domain 1 is Water deprivation, food deprivation, malnutrition; Domain 2 is Environmental challenge; Domain 3 is Disease, injury, functional impairment; Domain 4 is Behavioural, interactive restriction; and Domain 5 is Anxiety, fear, pain, distress, thirst, hunger and boredom. A proposal would be examined systematically in all these domains, and the degree of compromise in each rated on a five-step, non-numerical scale — O, A, B, C, X. Anxiety, fear, pain, distress, thirst, hunger and boredom arising from compromise in Domains 1–4, would be cumulated into Domain 5. The overall rating would commonly be that given to Domain 5, but if this were low or unknown, it would be given to the highest rating in the other domains. Each experimental group in a study would be rated so that compromise overall would neither be underestimated nor overestimated. Both the researcher and the animal ethics committee would rate each group in these terms.
Practical Difficulties in Balancing Harms and Benefits in the Modern Use of Laboratory Animals: Biotechnology
Authors: de Cock Buning Tjard
Published: April 2019

Every ethical review process presupposes a set of ethical criteria, proper procedures and a consistent legal framework. The Netherlands became unique in 1996, by the decision of the Parliament to start a national ethical review procedure regarding the ethical aspects of genetic modification of animals ( Animal and Biotechnology Decree). Due to the policy of public disclosure of all relevant documents and providing a legal trajectory for public objections, extensive ethical arguments pro and con are well documented. This paper lists and analyses the misconceptions when one tries to apply the “classical” ethical framework of harms and benefits to the new designs of animal experiments in the context of biotechnology. For instance, what is the object? The foster mothers, the manipulated egg cell, the malformed embryo, the pups dying before weaning or the adult mice in the experiments? The public considers the whole methodology ethically relevant. Most researchers tend to check out only “consciously” experienced harm, which implies only autonomously living mice and not a cell or an embryo. And, what is the domain of benefits? An exciting construct to be incorporated in the genome, the healthy birth of a surviving knock out, or a successful experiment with adult genetically modified mice? In the same way, the question for relevant alternatives in the “classic” sense of the Three Rs seems to shift completely to a discussion about goals in the far future. Is this modification absolutely necessary, or are the goals also reached by research strategies without genetic modification?
Biology Teachers’ Attitudes to Dissection and Alternatives
Authors: King Lesley A. , Ross Cheryl L. , Stephens Martin L. , Rowan Andrew N.
Published: April 2019

A survey of 5000 American middle and high school level biology teachers was completed to assess attitudes and classroom practice relating to dissection and alternative teaching methods. A preliminary sample of 494 respondents revealed that 79% of teachers used dissection to teach biology. While 72% believed that dissection was an important part of the curriculum, 17% disagreed; 69% considered dissection to be an essential hands-on activity. While 31% believed that alternatives were as good as dissection for teaching anatomy and physiology, 55% disagreed. The primary reason given for continuing dissection, rather than exclusively using alternatives, was the hands-on aspect of dissection (69%). While the majority (66%) of biology teachers favoured student choice between dissection and other learning methods, 20% disagreed. Although the effectiveness of alternative methods has been documented, and ethical arguments against dissection have been advanced, the mainstream introduction of humane alternatives to dissection requires a transformation of the beliefs, experience and practice of biology teachers.
Uses of Animals and Alternatives in Pre-college Education in the United States: Need for Leadership on Educational Resources and Guidelines
Authors: Hart Lynette A. , Wood Mary W. , Massey Ana , Smith Martin
Published: April 2019

Throughout pre-college education in the United States, animals, animal specimens and animals as a topic are used as teaching resources. Residential or visiting pets play a role in humane education or cross-curricular instruction. Teachers acquire and utilise non-living specimens gathered from various sources. Field trips often are oriented around animals. Elementary school animal use is largely observational; in intermediate grades, animal dissection may be featured in general science instruction. At the higher grade levels, animals may be used in science fair projects in an experimental sense. Particularly appealing to teachers is that throughout these uses, animals motivate students. The uses of animals in classrooms reflect the teachers’ interests. In the United States, the use of animals in pre-college instruction is not regulated, leading sometimes to inappropriate use. At the University of California, Davis, a pilot programme is assessing curricula, resources and a model prototype to provide administrative guidance on the use of animals in pre-college education. Appropriate animal use would be enhanced by: providing information resources with efficient links to obtaining educational materials; and establishing guidelines for animal use in pre-college education to ensure the animals’ welfare. A web-based tool provides access to resources ( www.vetmed.ucdavis.edu/Animal_Alternatives/main.htm ).
Our Experiences with Development of Digitised Video Streams and Their Use in Animal-free Medical Education
Authors: Červinka Miroslav , Červinková Zuzana , Novák Jan , Spicák Jan , Rudolf Emil , Peychl Jan
Published: April 2019

Alternatives and their teaching are an essential part of the curricula at the Faculty of Medicine. Dynamic screen-based video recordings are the most important type of alternative models employed for teaching purposes. Currently, the majority of teaching materials for this purpose are based on PowerPoint presentations, which are very popular because of their high versatility and visual impact. Furthermore, current developments in the field of image capturing devices and software enable the use of digitised video streams, tailored precisely to the specific situation. Here, we demonstrate that with reasonable financial resources, it is possible to prepare video sequences and to introduce them into the PowerPoint presentation, thereby shaping the teaching process according to individual students’ needs and specificities.
The Psychological Effects on Students of Using Animals in Ways that They See as Ethically, Morally or Religiously Wrong
Authors: Capaldo Theodora
Published: April 2019

Studies indicate that more students are opposing the use of invasive procedures on animals. When students are forced to use animals in ways that they view as harmful, painful, stressful or lethal to the animals, several reactions may occur. Such students may suffer psychological trauma as the result of seeing themselves or others engaged in behaviour that they find ethically objectionable. Their cognitive abilities may become impaired, resulting in less learning. They may withdraw and lose interest in science when not given the option to conscientiously object. Students — even those who believe they are willing participants — can become desensitised and may develop a utilitarian view of animals, thereby diminishing their capacity for compassion and ethical decision-making. Qualified, compassionate people — especially women — often decide to end their career in science rather than compromise their values. This loss contributes to the gender gap in science and to individuals feeling disappointed and derailed in their career aspirations. The psychological consequences to individuals combined with social, environmental, educational and scientific consequences present a compelling argument for the 100% replacement of the harmful use of animals in education.
Incorporating Animal Alternatives in a Training Programme in Laboratory Animal Care and Use
Authors: Walshaw Sally
Published: April 2019

Through the years, scientists and technicians, within their laboratories, trained individuals new to animal research. In the mid-1980s, new and revised laws/guidelines for animal research, including training, were instituted. The process of training scientists, technicians and students is recognised as essential for animal welfare and good science. Under the current and future revisions of the mandates, training must include specific topics. Training must include details of various regulations and guidelines, proper handling and care for animals, proper pre-procedural and post-procedural care of animals, aseptic surgical methods and procedures, proper use of anaesthetics, analgesics and tranquilisers, methods whereby deficiencies in animal care and treatment are reported, and a search for alternatives to the use of animals. Training individuals in clinical techniques (injection, blood collection, anaesthesia, etc.) is critical for humane treatment of animals, safety for the trainee and reliable animal data for the scientific project. Written materials about the clinical techniques should be provided at the training session, and training records must be kept.
“Statistics” is not a Sausage Machine: A Statistician's Viewpoint and Some Comments on Experimental Design
Authors: Das Rose E. Gaines
Published: April 2019

Any experiment involving the use of animals which is not well-planned, meticulously carried out, and scrupulously analysed, is unethical. Planning, or good experimental design, followed by analysis appropriate for the design, will help to ensure the optimal use of animals. Thus, collaboration between biologist and statistician, especially at the planning and analysis stages, is one of the best ways of achieving an ethical and successful experiment. However, genuine communication is necessary for any collaboration, and this requires time and patience, on the part of both biologist and statistician. Although the three fundamental principles of experimental design, replication, randomisation and local control, are straightforward in theory, there is substantial scope for misunderstanding and misinterpretation in practice. Each experiment presents unique and interacting biological and statistical problems, and both the right design and the correct analysis should be decided on a case-by-case basis.
A Statistical Evaluation of the Fixed Dose Procedure
Authors: Stallard Nigel , Whitehead Anne
Published: April 2019

The Fixed Dose Procedure (FDP) was first proposed in 1984 by the British Toxicology Society, as an alternative to the conventional LD50 test (OECD Test Guideline 401), for determining acute oral toxicity. The FDP used fewer animals and caused less suffering than the LD50 test, and provided information on acute toxicity which allowed substances to be classified according to the European Union hazard classification system. In 1992, the FDP was introduced as OECD Test Guideline 420. In 1999, as part of an initiative to phase out Test Guideline 401, a review of the FDP was undertaken. The aim of the review was to provide further reductions and refinements, and classification according to the criteria of the Globally Harmonised Hazard Classification and Labelling Scheme. The revised FDP was adopted by the OECD in 2001. This article concerns the development and revision of the FDP. It illustrates how statistical modelling and simulation can be used to increase the efficiency of a test procedure and reduce the number of animals needed for an in vivo validation of the procedure.
The Role of the Statistician in the Scottish Agricultural and Biological Research Institutes
Authors: McKendrick Iain J.
Published: April 2019

Several of the Scottish Agricultural and Biological Research Institutes carry out research on domestic animal health and welfare. Statistical services are provided by Biomathematics & Statistics Scotland, a sister research organisation. At one of these institutes, a statistician has been an integral member of the animal experiments and ethics committee for over 10 years, and each animal experiment is examined by the committee statistician as part of the review process. This paper will describe this review process, and then discuss those areas in which statistical advice has had most impact in the reduction of animal numbers. It is suggested that most benefit does not come from simple sample-size calculations, but rather from the application of the principles of good experimental design and close collaboration between the scientist and the statistician in the design and analysis of experiments. The final conclusion is that scientists welcome constructive, long-term statistical input, although budgetary issues can prove to be a barrier.
Improving Design and Analysis of Research: Lessons from Clinical Research
Authors: Altman Douglas G.
Published: April 2019

There is a clear need to optimise the use of animals in research and to ensure that the studies that are done make a worthwhile contribution to scientific knowledge. Research that has an inappropriate study design, or is improperly analysed or interpreted, may mislead and is not defensible. Published reviews of the quality of design and analysis in clinical studies are used to see what lessons can be drawn in respect of animal research. The need for clear and full reporting of research is discussed.
Reduction of Numbers of Animals Used in the Quality Control of Biologicals
Authors: Hendriksen Coenraad F.M.
Published: April 2019

Laboratory animals are used for the quality control of vaccines. In particular, the potency testing of batches of inactivated vaccine requires large numbers of animals. The possibilities for reduction have been evaluated, and the results are summarised in this paper. Several approaches were studied, including the retrospective analysis of test data, with the objectives of determining the minimum number of animals required per vaccine dilution group, and evaluating the feasibility of a single-dose potency test. Other studies focused on the development of serology-based models and the use of genetically uniform animals. Based on the outcome of these studies, a substantial reduction in the number of animals used for the potency testing of toxoid vaccines has been achieved or will be achieved in the near future. As reduction alternatives can generally be explored in a relatively simpler and less time-consuming way than replacement alternatives, more emphasis should be placed on reduction strategies than at present.
The Choice of Animal Model and Reduction
Authors: Festing Michael F.W.
Published: April 2019

Careful choice of the animal model is essential, if research is to be conducted efficiently, by using the minimum number of animals in order to provide the maximum amount of information. Inbred strains of rodents provide an excellent way of controlling and investigating genetic variation in characters of interest and in response to experimental treatments. Outbred stocks, in which genetic and non-genetic factors are inextricably mixed, are much less suitable, because random and uncontrolled genetic variation tends to obscure any treatment responses. In some cases, the use of inbred strains has led to major advances in scientific understanding. The specific example given here is in the understanding of host–parasite relationships but, more generally, inbred strains have been of critical importance in research which has resulted in the award of at least 17 Nobel prizes. And yet, despite the extensive literature on the properties and scientific value of inbred strains, many scientists continue to use outbred stocks in the mistaken belief that the use of such animals will, in some mysterious way, make their research more applicable to humans. There is really no evidence that this is so, and there is much evidence that the use of inbred strains has been highly successful in many disciplines.
Training for Reduction in Laboratory Animal Use
Authors: Nevalainen Timo
Published: April 2019

Reduction, refinement and replacement of laboratory animals wherever possible, are the guiding principles of the Federation of European Laboratory Animal Science Associations (FELASA) education guidelines. Of these, reduction is probably the least understood. Reduction and refinement are dependent upon each other. In the reduction “axis”, there is a window of appropriate numbers of animals; too few and the experiment will lack power — too many and animals will be wasted. Reduction must be understood as the appropriate number of animals required for each experiment. The refinement “axis” is more straightforward. Better welfare is always desirable. Any factor can interfere with a study in two ways. If it changes the mean, this may not be serious, because it should do so in all groups. If it causes a change in variation, then this is far more troublesome, because this is bound to alter the appropriate number of animals for an experiment. Scientists are definitely concerned about the variation of the characters that they are working with. It is obvious that changes in variation may be study-specific, which makes the formulation of overall guidelines difficult. Indeed, instead of trying to assess the impact of housing and procedures on every possible character, it could be more productive to look at the effects of welfare indicators on variation, with the understanding that low variation here is likely to be reflected by low variation in most other characters, while aiming to achieve the most uniform welfare of the animals in the study.
Assessment of the Eye Irritating Properties of Chemicals by Applying Alternatives to the Draize Rabbit Eye Test: The Use of QSARs and In Vitro Tests for the Classification of Eye Irritation
Authors: Gerner Ingrid , Liebsch Manfred , Spielmann Horst
Published: April 2019

Huggins has reported on the current situation relating to the development of alternatives to the Draize eye irritation test with rabbits, and an ECVAM Working Group have reviewed the efforts needed in order to replace this animal test within the next 10 years by using the results of non-animal assessment methods. Our report reviews regulatory experience gained over the last 20 years with the EU chemicals notification procedure with respect to the assessment of eye lesions observed in Draize tests. The nature of eye lesions and their importance for classification and labelling of possible hazards to human eyes are evaluated and discussed, with a view to promoting the development of specific in vitro assays which are able to discriminate between eye damage, moderate eye irritation, and minor irritation effects which are completely reversible within a few days. Structural alerts for the prediction of eye irritation/corrosion hazards to be classified and labelled according to international classification criteria, are presented, which should be validated in accordance with internationally agreed (OECD) principles for (Q)SAR system validation. Physicochemical limit values for prediction of the absence of any eye irritation potential relevant for human health can make available a definition of the applicability domains of alternative methods developed for the replacement of the Draize eye irritation test.
Alopecia Scoring: The Quantitative Assessment of Hair Loss in Captive Macaques
Authors: Honess Paul E. , Gimpel Jessica L. , Wolfensohn Sarah E. , Mason Georgia J.
Published: April 2019

Many captive animals show forms of pelage loss that are absent in wild or free-living con-specifics, which result from grooming or plucking behaviours directed at themselves or at other individuals. For instance, dorsal hair loss in primates such as rhesus macaques ( Macaca mulatta) in research facilities, results from excessive hair-pulling or over-grooming by cage-mates. This behaviour appears to be associated with stress, and is controllable to some extent with environmental enrichment. Quantifying alopecia in primates (as in many species) is therefore potentially useful for welfare assessment. A simple system for scoring alopecia was developed and its reliability was tested. Study 1 showed high interobserver reliability between two independent scorers in assessing the state of monkeys’ coats from photographs. Study 2 showed that there were no significant differences between the scores derived from photographs and from direct observations. Thus, where hair loss due to hair pulling exists in captive primates, this scoring system provides an easy, rapid, and validated quantitative method, for use in assessing the success of attempts to reduce it via improved husbandry. In the future, such scoring systems might also prove useful for quantifying barbering in laboratory rodents.
Animal Carcinogenicity Studies: 3. Alternatives to the Bioassay
Authors: Knight Andrew , Bailey Jarrod , Balcombe Jonathan
Published: April 2019

Conventional animal carcinogenicity tests take around three years to design, conduct and interpret. Consequently, only a tiny fraction of the thousands of industrial chemicals currently in use have been tested for carcinogenicity. Despite the costs of hundreds of millions of dollars and millions of skilled personnel hours, as well as millions of animal lives, several investigations have revealed that animal carcinogenicity data lack human specificity (i.e. the ability to identify human non-carcinogens), which severely limits the human predictivity of the bioassay. This is due to the scientific inadequacies of many carcinogenicity bioassays, and numerous serious biological obstacles, which render profoundly difficult any attempts to accurately extrapolate animal data in order to predict carcinogenic hazards to humans. Proposed modifications to the conventional bioassays have included the elimination of mice as a second species, and the use of genetically-altered or neonatal mice, decreased study durations, initiation–promotion models, the greater incorporation of toxicokinetic and toxicodynamic assessments, structure-activity relationship (computerised) systems, in vitro assays, cDNA microarrays for detecting changes in gene expression, limited human clinical trials, and epidemiological research. The potential advantages of non-animal assays when compared to bioassays include the superior human specificity of the results, substantially reduced time-frames, and greatly reduced demands on financial, personnel and animal resources. Inexplicably, however, the regulatory agencies have been frustratingly slow to adopt alternative protocols. In order to decrease the enormous cost of cancer to society, a substantial redirection of resources away from excessively slow and resource-intensive rodent bioassays, into the further development and implementation of non-animal assays, is both strongly justified and urgently required.