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 If refinement is not actively sought, outdated and unnecessarily invasive techniques may not be replaced by better methods as they become available, and thus outdated information is passed down to the next generation, causing perpetuation of old-fashioned methods.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.

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.

Case report of a possible familial predisposition to metabolic bone disease in juvenile rhesus macaques.

Deficiencies of dietary calcium and/or vitamin D will cause hypocalcaemia, leading to metabolic bone disease. The disease commonly affects young rapidly growing animals and this is a report of the condition in a colony of rhesus macaques (Macaca mulatta). A clinical problem of metabolic bone disease was seen in 1993, when it was treated and resolved satisfactorily. However it recurred in 1999 following changes in management and husbandry of the colony, at which time the clinical manifestations were more serious. The animals had bowed tibia, fibula, radius and ulna and enlarged epiphyses, were reluctant to climb and jump, had a 'hopping' gait and poor growth. The syndrome had a multifactorial aetiology involving a combination of staff and management changes, a borderline nutritional deficit, a lack of daylight for production of vitamin D, and a possible familial predisposition.

Interpretation of serological test results for simian herpes B virus.

In 1992 an annual Simian herpes B virus (BV) screening programme for an experimental group of macaque monkeys (Macaca mulatta and Macaca fascicularis) was initiated with the aim of establishing a specific pathogen free (SPF) colony. In June 1999 one animal was found to be unexpectedly BV positive (non-negative). The investigation of this result highlights some of the issues and difficulties that may be encountered in such a programme.

Toward developing a genome-wide microsatellite marker set for linkage analysis in the rhesus macaque (Macaca mulatta): identification of 76 polymorphic markers.

Linkage analysis can be problematic in humans because of the lack of large, multigenerational pedigrees and the difficulties in obtaining phenotypic data on all family members. In contrast, large, captive colonies of rhesus macaque are a potentially valuable resource for linkage studies because detailed phenotypic and genealogical data are kept, inbreeding is avoided, and DNA samples can usually be obtained. Microsatellite marker sets for genome-wide screening are available in a number of species, but not for the rhesus macaque. We tested primers to 400 human microsatellite markers from a genome-wide mapping set using DNA from nine unrelated female rhesus macaques. We found that 76 (19%) of the primers amplified a polymorphic product using the standard protocols for human DNA. The average heterozygosity of the markers in humans was 0.80, compared to 0.65 in the rhesus macaques. This study provides preliminary data, which could be used toward the development of a linkage mapping set in this species. There would be a need, however, to confirm the Mendelian inheritance of the markers.

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.

Anaemia in housed newborn lambs.

The mean (+/- sd) packed cell volume (PCV) of the lambs in a flock mated by 'out-of-season' breeding methods and housed throughout the last six weeks of pregnancy and the whole of the suckling period declined to 23.3 +/- 3.34 per cent when the lambs were a month old, but recovered spontaneously to reach values within the normal range (30 to 35 per cent) before they were weaned at two months. Clinical signs of anaemia were observed in only a few lambs at three weeks old but the PCV values of all the lambs were significantly lower at this time than at weaning. There was no relationship between the PCV values during the two to four weeks after birth and the growth rate of the lambs. The administration of 200 mg iron, as iron dextran, by intramuscular injection to the newborn lambs prevented the decrease in PCV, but had no measurable effect on the health or growth rate of the lambs throughout the suckling period.