Nuno H. Franco

Animal Experiments in Biomedical Research: A Historical Perspective

Simple Summary This article reviews the use of non-human animals in biomedical research from a historical viewpoint, providing an insight into the most relevant social and moral issues on this topic across time, as well as to how the current paradigm for ethically and publically acceptable use of animals in biomedicine has been achieved. Abstract The use of non-human animals in biomedical research has given important contributions to the medical progress achieved in our day, but it has also been a cause of heated public, scientific and philosophical discussion for hundreds of years. This review, with a mainly European outlook, addresses the history of animal use in biomedical research, some of its main protagonists and antagonists, and its effect on society from Antiquity to the present day, while providing a historical context with which to understand how we have arrived at the current paradigm regarding the ethical treatment of animals in research.

Scientists and the 3Rs: attitudes to animal use in biomedical research and the effect of mandatory training in laboratory animal science

The 3Rs principle of replacement, reduction, and refinement has increasingly been endorsed by legislators and regulatory bodies as the best approach to tackle the ethical dilemma presented by animal experimentation in which the potential benefits for humans stand against the costs borne by the animals. Even when animal use is tightly regulated and supervised, the individual researcher’s responsibility is still decisive in the implementation of the 3Rs. Training in laboratory animal science (LAS) aims to raise researchers’ awareness and increase their knowledge, but its effect on scientists’ attitudes and practice has not so far been systematically assessed. Participants (n = 206) in eight LAS courses (following the Federation of European Laboratory Animal Science Associations category C recommendations) in Portugal were surveyed in a self-administered questionnaire during the course. Questions were related mainly to the 3Rs and their application, attitudes to animal use and the ethical review of animal experiments. One year later, all the respondents were asked to answer a similar questionnaire (57% response rate) with added self-evaluation questions on the impact of training. Our results suggest that the course is effective in promoting awareness and increasing knowledge of the 3Rs, particularly with regard to refinement. However, participation in the course did not change perceptions on the current and future needs for animal use in research.

How “Humane” Is Your Endpoint?—Refining the Science-Driven Approach for Termination of Animal Studies of Chronic Infection

potential human benefits, but if accepting the assumption that human benefits can offset animal suffering, it still needs to be argued that the same benefits could not be achieved with less negative effects on animal welfare. Reducing the animal welfare problems associated with research (‘‘refinement’’ [1]) is therefore crucial in order to render animal-based research less of an ethical problem and to assure public trust in research. Studies that are designed to measure time of death or survival percentages present a particularly challenging situation in which at least some of the animals are made to die from the disease. These studies are frequent in experimental research on severe infections. The scientific community, industry, and regulatory authorities have responded to the ethical concerns over studies in which animals die from severe disease by developing new policies and guidelines for the implementation of humane endpoints as a key refinement measure (e.g., [2–4]). The most widely used definition considers a humane endpoint to be the earliest indicator in an animal experiment of severe pain, severe distress, suffering, or impending death [5], underlining that ideally such indicators should be identified before the onset of the most severe effects. Euthanizing animals, rather than awaiting their ‘‘spontaneous’’ death, is important to avoid unnecessary suffering in studies in which data on survival is thought to be required for scientific or legal reasons. However, several questions remain open regarding how humane endpoints are to be applied to address real animal welfare problems. We used TB experiments in mice as a case study to highlight the potential to establish biomarkers of disease progress that can replace survival time as a measure of disease severity.

Animal Welfare in Studies on Murine Tuberculosis: Assessing Progress over a 12-Year Period and the Need for Further Improvement

There is growing concern over the welfare of animals used in research, in particular when these animals develop pathology. The present study aims to identify the main sources of animal distress and to assess the possible implementation of refinement measures in experimental infection research, using mouse models of tuberculosis (TB) as a case study. This choice is based on the historical relevance of mouse studies in understanding the disease and the present and long-standing impact of TB on a global scale. Literature published between 1997 and 2009 was analysed, focusing on the welfare impact on the animals used and the implementation of refinement measures to reduce this impact. In this 12-year period, we observed a rise in reports of ethical approval of experiments. The proportion of studies classified into the most severe category did however not change significantly over the studied period. Information on important research parameters, such as method for euthanasia or sex of the animals, were absent in a substantial number of papers. Overall, this study shows that progress has been made in the application of humane endpoints in TB research, but that a considerable potential for improvement remains.

Children’s attitudes towards animals: evidence from the RODENTIA project

The instructional use of animals is a popular strategy to engage students with science, enhance their motivation, and promote values such as respect, tolerance, and empathy for all living beings. Although these beneficial outcomes are widely acknowledged, research has not provided reliable indicators of their efficiency. Therefore, it is essential to broaden the studies focused on the use of animals in education. In this regard, it becomes necessary to understand students’ attitudes towards animals. This paper presents data on the attitudes of primary school children towards the humane treatment of animals. It follows the implementation of a longitudinal project based on the concept of ‘classroom pet’, aimed at fostering the development of scientific reasoning competencies and positive attitudes towards animals. To assess the project’s efficacy, a methodology combining quantitative and qualitative assessment approaches was outlined. The study involved 43 students, aged 8–10, from two fourth-grade classes of the same school. Findings concerning how children’s attitudes towards different animals and animal uses are modulated as the result of an animal-based educational intervention and naturally throughout their maturation are discussed. This study provides relevant information for the development and evaluation of humane educational programmes.

Welfare and Quantity of Life

1. ABSTRACT The Animal welfare science is mostly focused on evaluating and improving the quality of life of animals that actually exist. This leaves out a range of ethically relevant issues regarding the quantity of life – in terms of number of animals living and the longevity of each animal. In many cases quantity and quality are related, and often there is a tension between the two. In this chapter, we develop a discussion around four practical cases presenting quality-quantity dilemmas: a) the issue of dairy cow longevity, b) the early slaughter of male dairy calves, c) the killing of newly-hatched male layer chicks and d) the conflict between reduction and refinement in animal research. The practical, economic and animal welfare aspects characterizing each case are presented together with relevant stakeholders’ perspective. We discuss the cases in light of the most relevant currents of thought in animal ethics, highlighting the main values at stake and which possible solutions may be sought according to each perspective.

A Critical Look at Biomedical Journals’ Policies on Animal Research by Use of a Novel Tool: The EXEMPLAR Scale

Simple Summary Biomedical journals have the responsibility to promote humane research. To gauge and evaluate journal policies on animal research, the EXEMPLAR—For “Excellence in Mandatory Policies on Animal Research”—scale is presented and applied to evaluate a sample of 170 biomedical journals, providing an overview of the current landscape of editorial policies on the ethical treatment of animals. Abstract Animal research is not only regulated by legislation but also by self-regulatory mechanisms within the scientific community, which include biomedical journals’ policies on animal use. For editorial policies to meaningfully impact attitudes and practice, they must not only be put into effect by editors and reviewers, but also be set to high standards. We present a novel tool to classify journals’ policies on animal use—the EXEMPLAR scale—as well as an analysis by this scale of 170 journals publishing studies on animal models of three human diseases: Amyotrophic Lateral Sclerosis, Type-1 Diabetes and Tuberculosis. Results show a much greater focus of editorial policies on regulatory compliance than on other domains, suggesting a transfer of journals’ responsibilities to scientists, institutions and regulators. Scores were not found to vary with journals’ impact factor, country of origin or antiquity, but were, however, significantly higher for open access journals, which may be a result of their greater exposure and consequent higher public scrutiny.

Killing animals as a necessary evil? The case of animal research

This chapter addresses the question of killing animals in research, primarily from a moral perspective, but also taking into account some of the practical and scientific considerations with moral consequences in this context. We start by exploring in which situations animals are killed in research and whether these are always inevitable, analysing re-use and re-homing of animals as potential alternatives. We then discuss for whom – and under what circumstances -killing matters, considering situations where there may be a conflict between the wish to avoid killing and that to avoid suffering, and further take humananimal interactions into account. We argue that, although there are relevant practical, scientific and ethical arguments favouring the euthanasia of animals in most research contexts, there is a potential for rehabilitating more animals than is currently the practice.

Eye, body or tail? Thermography as a measure of stress in mice

Infrared thermography has been suggested as a non-invasive, objective tool to evaluate animal welfare. In this study, we investigated: 1) how body temperature, measured through thermal imaging, is affected by different mild stressors frequently experienced by laboratory mice; 2) which methodology to use for assessing temperature variations with infrared thermography; 3) whether the chosen stressors cause anxiety in mice. Eighty C57BL/6 male mice were included in the study. The mice were allocated to either a control group or one of three groups being subjected to a mild stressor once daily for 4 days: 1) anaesthesia with isoflurane for 10 min; 2) handling by scruffing; 3) intraperitoneal injection of 0.2 ml 0.9% saline. On all four intervention days, thermal images were obtained in all groups and all animals were assessed for fur status and body weight. On day five, all animals were tested in the elevated-plus-maze for 5 min. From the thermal images, the maximum eye temperature, the maximum tail base temperature and the average body temperature were obtained. Ten minutes of anaesthesia with isoflurane led to a decrease in maximum eye temperature, average body temperature and maximum tail base temperature. The animals recovered from this drop in temperature within 10 min. No drop in temperature was seen after scruffing or intraperitoneal injection of saline. Based on the number of missing values, intra-rater and inter-rater agreement, the average body temperature was found most ideal for measuring body temperature variations in mice. Finally, the elevated plus maze did not reveal any differences in anxiety between the groups and the body weight did not decrease at any time point during the study.

Researchers’ attitudes to the 3Rs—An upturned hierarchy?

Animal use in biomedical research is generally justified by its potential benefits to the health of humans, or other animals, or the environment. However, ethical acceptability also requires scientists to limit harm to animals in their research. Training in laboratory animal science (LAS) helps scientists to do this by promoting best practice and the 3Rs. This study evaluated scientists’ awareness and application of the 3Rs, and their approach to other ethical issues in animal research. It was based on an online survey of participants in LAS courses held in eight venues in four European countries: Portugal (Porto, Braga), Germany (Munich, Heidelberg), Switzerland (Basel, Lausanne, Zurich), and Denmark (Copenhagen). The survey questions were designed to assess general attitudes to animal use in biomedical research, Replacement alternatives, Reduction and Refinement conflicts, and harm-benefit analysis. The survey was conducted twice: immediately before the course (‘BC’, N = 310) and as a follow-up six months after the course (‘AC’, N = 127). While courses do appear to raise awareness of the 3Rs, they had no measurable effect on the existing low level of belief that animal experimentation can be fully replaced by non-animal methods. Most researchers acknowledged ethical issues with their work and reported that they discussed these with their peers. The level of an animal’s welfare, and especially the prevention of pain, was regarded as the most pressing ethical issue, and as more important than the number of animals used or the use of animals as such. Refinement was considered more feasible than Replacement, as well as more urgent, and was also favoured over Reduction. Respondents in the survey reversed the ‘hierarchy’ of the 3Rs proposed by their architects, Russell and Burch, prioritizing Refinement over Reduction, and Reduction over Replacement. This ordering may conflict with the expectations of the public and regulators.