Sarah Wolfensohn

Evidence from rhesus macaques suggests that male coloration plays a role in female primate mate choice

Male animals of many species use conspicuous coloration to attract mates. Among mammals, primates possess the most brilliant secondary sexual coloration. However, whether colour plays a part in primate female mate choice remains unknown. Adult male rhesus macaques undergo a hormonally regulated increased reddening of facial and anogenital skin during their mating season. We experimentally investigated whether red male facial coloration is preferred by simultaneously presenting female rhesus macaques (n = 6) with computer-manipulated pale and red versions of 24 different male faces. The duration and direction of gaze were measured to discern visual preferences. Females exhibited preferences for the red versions of male faces. It is proposed that male coloration might provide a cue to male quality.

Refinements in husbandry, care and common procedures for non-human primates Ninth report of the BVAAWF/FRAME/RSPCA/UFAW Joint Working Group on Refinement

Preface Whenever animals are used in research, minimizing pain and distress and promoting good welfare should be as important an objective as achieving the experimental results. This is important for humanitarian reasons, for good science, for economic reasons and in order to satisfy the broad legal principles in international legislation. It is possible to refine both husbandry and procedures to minimize suffering and improve welfare in a number of ways, and this can be greatly facilitated by ensuring that up-to-date information is readily available. The need to provide such information led the British Veterinary Association Animal Welfare Foundation (BVAAWF), the Fund for the Replacement of Animals in Medical Experiments (FRAME), the Royal Society for the Prevention of Cruelty to Animals (RSPCA) and the Universities Federation for Animal Welfare (UFAW) to establish a Joint Working Group on Refinement (JWGR) in the UK. The chair is Professor David Morton and the secretariat is provided by the RSPCA. This report is the ninth in the JWGR series. The RSPCA is opposed to the use of animals in experiments that cause pain, suffering, distress or lasting harm and together with FRAME has particular concerns about the continued use of non-human primates. The replacement of primate experiments is a primary goal for the RSPCA and FRAME. However, both organizations share with others in the Working Group, the common aim of replacing primate experiments wherever possible, reducing suffering and improving welfare while primate use continues. The reports of the refinement workshops are intended to help achieve these aims. This report produced by the British Veterinary Association Animal Welfare Foundation (BVAAWF)/Fund for the Replacement of Animals in Medical Experiments (FRAME)/Royal Society for the Prevention of Cruelty to Animals (RSPCA)/Universities Federation for Animal Welfare (UFAW) Joint Working Group on Refinement (JWGR) sets out practical guidance on refining the husbandry and care of non-human primates (hereinafter primates) and on minimizing the adverse effects of some common procedures. It provides a valuable resource to help understand the physical, social and behavioural characteristics and needs of individual primates, and is intended to develop and complement the existing literature and legislative guidelines. Topics covered include refinements in housing, husbandry and common procedures such as restraint, identification and sampling, with comprehensive advice on issues such as primate communication, assessing and facilitating primate wellbeing, establishing and maintaining social groups, environmental and nutritional enrichment and animal passports. The most commonly used species are the key focus of this resource, but its information and recommendations are generally applicable to other species, provided that relevant individual species characteristics are taken into account.

A study of behavioural responses of non-human primates to air transport and re-housing:

More long-tailed macaques (Macaca fascicularis) than any other primate are imported into the UK for research, and journey times may be of up to 58 h. Whilst a number of studies have examined the stress associated with transport, these have typically involved laboratory rodents and livestock, and little is known of its effect on non-human primates. This paper reports the results of a study of behavioural changes in a group of long-tailed macaques transported by air from standard breeding conditions and then re-housed in standard laboratory primate conditions. The animals were studied prior to their departure, immediately after their arrival, and 3 weeks after that. Data were collected on individual time budgets using focal animal sampling and on hierarchy using a feeding trial. The data were analysed for changes in behavioural repertoires and for social perturbation that would be reflected in hierarchical changes. Changes in behaviour occurred which reflected heightened levels of stress in the study group. It was also clear that although there was some adjustment of behaviour, after an initial change on arrival at the new establishment, there was no return to levels observed at the breeding facility within the first month. This study demonstrates that, as a whole, the process of international air transport and re-housing in laboratory conditions may result in the compromising of the welfare of the study animals.

Guidelines for the veterinary care of laboratory animals: report of the FELASA/ECLAM/ESLAV Joint Working Group on Veterinary Care

Summary Veterinary professionals working in partnership with other competent persons are essential for a successful animal care and use programme. A veterinarians primary responsibilities are defined by their own professional regulatory bodies, but in this area of work there are further opportunities for contribution, which will assist in safeguarding the health and welfare of animals used in research. These guidelines are aimed not only at veterinarians to explain their duties, and outline the opportunities to improve the health and welfare of animals under their care, but also at employers and regulators to help them meet their responsibilities. They describe the desirability for postgraduate education towards specialization in laboratory animal medicine and detail the many competencies necessary to fulfil the role of the laboratory animal veterinarian. They detail the need for veterinary expertise to promote good health and good welfare of animals used in biomedical research during husbandry as well as when under experimental procedures. Regulatory and ethical aspects are covered as are the involvement of the veterinarian in education and training of others working in the animal care and use programme. Managerial aspects, including occupational health and safety, are also areas where the veterinarians input can assist in the successful implementation of the programme.

Refinement : promoting the three Rs in practice

Refinement of scientific procedures carried out on protected animals is an iterative process, which begins with a critical evaluation of practice. The process continues with objective assessment of the impact of the procedures, identification of areas for improvement, selection and implementation of an improvement strategy and evaluation of the results to determine whether there has been the desired effect, completing the refinement loop and resulting in the perpetuation of good practice. Refinements may be science-driven (those which facilitate getting high-quality results) or welfare-driven or may encompass both groups, but whatever the driver, refinements almost always result in benefits to both welfare and science. Refinements can be implemented in all aspects of animal use: improved methodology in invasive techniques, housing and husbandry, and even statistical analyses can all benefit animal welfare and scientific quality. If refinement is not actively sought, outdated and unnecessarily invasive techniques may not be replaced by better methods as they become available, and thus outdated information is passed down to the next generation, causing perpetuation of old-fashioned methods. This leads to a spiral of ignorance, leading ultimately to poor practice, poor animal welfare and poor-quality scientific data. Refinement is a legal and ethical requirement, yet refinements may not always be implemented. There are numerous obstacles to the implementation of refinement, which may be real or perceived. Either way, in order to take refinement forward, it is important to coordinate the approach to refinement, validate the science behind refinement, ensure there is adequate education and training in new techniques, improve liaison between users and make sure there is feedback on suitability of refinements for use. Overall, refinement requires a coordinated ongoing process of critical appraisal of practice and active scrutiny of resources for likely improvements. In the busy world of biomedical research, this process needs help. In order to develop these themes further, a workshop was held at the LASA Winter Meeting 2006, UK, to assist in identifying potential obstacles to refinement, and then to explore and develop strategies for overcoming these obstacles in key areas. A range of strategies appropriate to different circumstances was identified, which should facilitate the implementation of refinements.

Refinement of welfare through development of a quantitative system for assessment of lifetime experience

© 2015 Universities Federation for Animal Welfare.This paper proposes a system that uses intrinsic study data to provide a clear visualisation of the stresses involved during the animals life history that can be applied to all types of studies, even those not requiring invasive techniques. Thus, it provides an opportunity for researchers to identify and refine key events which impact on the welfare of an animal, and to explain clearly the totality of any necessary harms when justifying the research. Assessment of animal welfare depends on measurement of a number of parameters which will vary according to species, the animals environment and the scientific procedure, all of which are inter-related. Currently, there are few tools to assess the effects of lifetime events on welfare or even, in some cases, to recognise that they have an impact on the level of suffering. A matrix to assess the combined effects of environment, experimental and contingent events on welfare has been applied, retrospectively, to programmes of work involving macaques (Macaca mulatta and M. fascicularis). Lifetime records, available for animals from their birth in the breeding colony through to experimental use in vaccine efficacy evaluation studies, were analysed as a robust validation test for the assessment matrix and refinement of the way in which information on these events is captured. A meaningful assessment method is required prospectively for project licence applications and retrospectively for licence review or decisions on re-use. The analysis will provide information that would support the application of refinements that would optimally enhance the lives of experimental animals.

Report on primate supply for biomedical scientific work in the UK

A Working Party of the UK group of European Primate Resources Network (EUPREN) considered primate supply for scientific work in the UK. Through a questionnaire, which achieved a very good response, it obtained details of primate use, sources and breeding in the UK and it put forward options to ensure that animal welfare is the best possible whilst ensuring continued supply. The questionnaire showed that contract research laboratories and pharmaceutical companies use about 80% of the 4233 primates used annually at the moment, with the rest accounted for by academic establishments and public sector laboratories. Fifty-four per cent are cynomolgus macaques (Macaca fascicularis), of which nearly 90% are captive-bred outside the European Union (EU), the remainder being bred in the UK. Nearly 90% of cynomolgus macaques are used by only five institutions. Thirty-seven per cent of primates used are marmosets (Callithrix jacchus jacchus), all of which are bred in the UK. Most of the rest are rhesus macaques (Macaca mulatta), about half of which are captive-bred outside the EU, the other half being bred in the UK. Overall primate use has increased from about 3000 per year in 1990 and users predict that requirements for all species except baboons (Papio sp.) will be maintained or increase. Marmoset breeding in the UK is already closely matched to use, and it could be increased reasonably easily if necessary. Some of the existing breeding centres of macaques in the UK would be prepared to consider expanding to supply others, although investment and imported breeding stock would be needed and it is likely that a large investment would be needed to breed a significant fraction of the macaque use in the UK. A further problem is that the users of only about 10% of the cynomolgus macaques said that they could replace this species by rhesus macaques, which are easier to breed in the UK. The questionnaire showed that much of the use of macaques would be transferred to other countries equally remote from the natural source countries of the animals, if constraints on primate use became more severe in the UK. Users felt that it is unlikely that much of the work could be transferred to the natural source countries themselves. A review of the literature revealed a paucity of information on the effects of transport on primate welfare. The importance of obtaining this information before making decisions about alternative means of supply is stressed. Current schemes for the accreditation of primate breeders were reviewed. A list of options is presented for discussion. Users vary so much in their requirements that it is unlikely that one means of supply will be applicable to all. Animal welfare will benefit and supply will be more certain if cooperation between those concerned (preferably through the UK group of EUPREN) is maintained.

BRIEF REVIEW OF SCIENTIFIC STUDIES OF THE WELFARE IMPLICATIONS OF TRANSPORTING PRIMATES

The transportation of primates has become an important welfare issue and the outcome of the debate over its cost to the animal will have effects on the future of medical research using these species. There is a paucity of scientific studies on transport relating to primates and the need for gathering of further scientific evidence is highlighted.

Chloral Hydrate Is Not Acceptable for Anesthesia or Euthanasia of Small Animals

To the Editor:—A recent paper reported experiments, using laboratory rats, on the effect of intraamygdala infusion of a -aminobutyric acid type A antagonist on propofol-induced amnesia for inhibitory avoidance training, as well as on expression of activity-regulated cytoskeleton-associated protein in the hippocampus. This work may elucidate the neural mechanisms of the amnestic effects of propofol, as well as the neurobiological mechanisms of general anesthesia and memory more generally. The experiments reported in this paper required stereotaxic neurosurgery to implant cannulae aimed at the basolateral amygdala, and rats were subsequently euthanized for determination of activity-related cytoskeleton-associated protein levels or histologic verification of cannula placement. The authors used chloral hydrate in both procedures; for surgical anesthesia in the first and euthanasia in the second. Chloral hydrate is not a suitable drug in either case. Chloral hydrate is regarded by many to produce hypnosis and not anesthesia. It does not provide analgesia and causes marked respiratory depression at doses required for surgical anesthesia. Apart from its inadequate anesthetic properties, 20% chloral hydrate is extremely irritating and therefore unsuitable for intraperitoneal use. It is associated with ileus in rats, as well as peritonitis and gastric ulcers. Its use by intraperitoneal injection for survival surgery is not recommended. Thus, it is not the most refined choice of agent for the surgical procedure in which cannulae are chronically implanted to make drug infusions into the amygdala. The authors also used a higher dose of chloral hydrate for euthanasia. However, chloral hydrate is not an acceptable agent for euthanasia according to the guidelines of the American Veterinary Medical Association†; its use for this purpose has been proscribed for some time. There are no scientific justifications for using chloral hydrate for these experiments, as many other agents would be more suitable for both surgical anesthesia and euthanasia without interfering with the experimental endpoints. Indeed the chloral hydrate-induced hypoxemia which must occur during euthanasia as respiration becomes depressed†, may compromise the experimental aims in terms of measuring protein and messenger ribonucleic acid levels of an activityrelated protein. The noxious stimulus of an intraperitoneal irritant is not only inhumane, but if it leads to peritonitis the rats will be abnormal at the time of testing. It seems that chloral hydrate has traditionally been used to provide anesthesia where the avoidance of agents with known receptor interactions is desirable. But it is likely that chloral hydrate has unknown receptor interactions. Therefore choosing a different agent whose receptor interactions are better characterized could be beneficial, not only in terms of animal welfare but also in terms of data interpretation. The publication of this paper in ANESTHESIOLOGY concerns us, because the standard of laboratory animal anesthesia used in this research is not acceptable.

Developing a Collaborative Agenda for Humanities and Social Scientific Research on Laboratory Animal Science and Welfare

Improving laboratory animal science and welfare requires both new scientific research and insights from research in the humanities and social sciences. Whilst scientific research provides evidence to replace, reduce and refine procedures involving laboratory animals (the ‘3Rs’), work in the humanities and social sciences can help understand the social, economic and cultural processes that enhance or impede humane ways of knowing and working with laboratory animals. However, communication across these disciplinary perspectives is currently limited, and they design research programmes, generate results, engage users, and seek to influence policy in different ways. To facilitate dialogue and future research at this interface, we convened an interdisciplinary group of 45 life scientists, social scientists, humanities scholars, non-governmental organisations and policy-makers to generate a collaborative research agenda. This drew on methods employed by other agenda-setting exercises in science policy, using a collaborative and deliberative approach for the identification of research priorities. Participants were recruited from across the community, invited to submit research questions and vote on their priorities. They then met at an interactive workshop in the UK, discussed all 136 questions submitted, and collectively defined the 30 most important issues for the group. The output is a collaborative future agenda for research in the humanities and social sciences on laboratory animal science and welfare. The questions indicate a demand for new research in the humanities and social sciences to inform emerging discussions and priorities on the governance and practice of laboratory animal research, including on issues around: international harmonisation, openness and public engagement, ‘cultures of care’, harm-benefit analysis and the future of the 3Rs. The process outlined below underlines the value of interdisciplinary exchange for improving communication across different research cultures and identifies ways of enhancing the effectiveness of future research at the interface between the humanities, social sciences, science and science policy.