Celebrating Women in Science: Pioneering Contributions to Animal Behaviour and Welfare.

In the scientific domain, women have historically demonstrated their dedication, intellect, and innovative input in relation to animal behaviour and welfare [...].

Toward Global Harmonization of Training and Certification of Specialists in Laboratory Animal Veterinary Medicine.

Laboratory animal medicine (LAM) is a corner stone of animal-based research and has been a veterinary specialty for over 60 y. Today 5 Colleges of LAM (American, European, Japanese, Korean, and Indian) that certify specialists (Diplomates) in LAM are members of the International Association of Colleges of LAM (IACLAM). Goals of IACLAM are to support the development of new Colleges of LAM, to harmonize expectations for the knowledge and skills of newly certified LAM Diplomate, and to harmonize the standards (best practices) for training and examination of candidates among the member Colleges. IACLAM recently conducted an in-depth review and comparison of oversight, training, credentialing, and examination standards in the 5 Colleges as part of an initiative to create a framework for harmonization and consistency for these activities across the 5 Colleges. The process has led to an agreement on recommendations for knowledge and skill requirements for a newly certified Diplomate, as described by each College in a detailed role delineation document (RDD). The RDD is based on task analyses of the work responsibilities of laboratory animal veterinary Diplomates. This agreement is an important step toward the goal of global harmonization of LAM Diplomate training. Further efforts are planned for areas such as training, research, publication, and examination. This paper describes the role and content of the RDD and lists the differences and similarities among the RDDs of 5 Colleges of LAM.

Culture of Care Enhancement in Egypt: The Impact of Laboratory Animal Science Training on Participants' Attitudes.

Cairo University was the first academic institution in Egypt to establish an Institutional Animal Care and Use Committee (IACUC), as mandated by the World Organisation for Animal Health (OIE). Animal-based research should be performed in accordance with international regulations to monitor the humane care and use of the laboratory animals. Until 2018, the formal training of researchers in the appropriate and correct methods of animal handling during sampling and administration, as well as their husbandry demands, was an uncommon practice in Egypt. In 2018, the Egyptian Association for Animal Research Advancement (EAARA) organised the first international course in laboratory animal science (LAS), in collaboration with Utrecht University (The Netherlands) and the Faculty of Science, Cairo University, to raise researchers' awareness and increase their knowledge of the principles that govern the humane use and care of laboratory animals. A total of 26 researchers from a number of fields (veterinary medicine, dentistry, science, medicine, pharmacy and agriculture) enrolled in the course. In the responses to the post-course questionnaire, 24 (92.3%) participants stated that the principles of animal welfare (Three Rs) were well explained. In addition, 18 (69%) participants found that the course improved their skills in animal sampling and handling. Of the 26 participants, 22 (84.6%) became aware of their responsibility towards their experimental animals and agreed that the different methods of euthanasia were well explained. In conclusion, the general assessment of the course revealed a positive outcome regarding the culture of animal care; the course was repeated a year later, and several participants were enlisted as trainers in this second course.

Welfare Impact of Carbon Dioxide Euthanasia on Laboratory Mice and Rats: A Systematic Review.

Background: There has been increased concern about the suitability of CO2 as a method for euthanasia of laboratory mice and rats, including the potential discomfort, pain or distress that animals may experience prior to loss of consciousness; time to loss of consciousness; best methods for use of CO2; and the availability of better alternatives. These discussions have been useful in providing new information, but have resulted in significant confusion regarding the acceptability of CO2 for rodent euthanasia. In some cases, researchers and veterinarians have become uncertain as to which techniques to recommend or use for euthanasia of laboratory mice and rats. Methods: The International Association of Colleges of Laboratory Animal Medicine (IACLAM) convened a taskforce to examine the evidence for adverse welfare indicators in laboratory rats and mice undergoing CO2 euthanasia using a SYRCLE-registered systematic review protocol. Of 3,772 papers identified through a database search (PubMed, Web of Science, CAB Direct, Agricola, and grey literature) from 1900 to 2017, 37 studies were identified for detailed review (some including more than one species or age group), including 15 in adult mice, 21 in adult rats, and 5 in neonates of both species. Experiments or reports were excluded if they only assessed parameters other than those directly affecting animal welfare during CO2 induction and/or euthanasia. Results: Study design and outcome measures were highly variable and there was an unclear to high risk of bias in many of the published studies. Changes in the outcome measures evaluated were inconsistent or poorly differentiated. It is likely that repeated exposures to carbon dioxide inhalation are aversive to adult rats and mice, based on avoidance behavior studies; however, this effect is largely indistinguishable from aversion induced by repeated exposures to other inhalant anesthetic gasses. Conclusion: There is insufficient evidence to permit an unbiased assessment of the effect of CO2 inhalation during euthanasia on welfare indicators in laboratory mice and rats. Additional well-designed, unbiased, and adequately powered studies are needed to accurately assess the welfare of laboratory mice and rats undergoing euthanasia via CO2 gas.

Reference values for selected hematological, biochemical and physiological parameters of Sprague-Dawley rats at the Animal House, Faculty of Medicine, University of Colombo, Sri Lanka.

BACKGROUND: Lack of available reference values in a research setting under local conditions can be a drawback for beginners, as the accuracy of data from control samples cannot be checked at the beginning of a research project. This affects comparisons with data from test samples. To avoid these complications in their research projects, beginners tend to have a greater number of animals in the control group compared to test groups in order to have control group measurements within 2 SDs of the mean. METHODS: As non-availability of reference values was a long-felt need, the described project was conducted in order to establish a reference database for selected haematological, biochemical and physiological parameters using apparently healthy Sprague-Dawley rats bred in the Animal House of Faculty of Medicine, University of Colombo (UCFM). RESULTS: Differences in mean values of packed cell volume (PCV), mean corpuscular volume (MCV) and mean corpuscular hemoglobin concentration (MCHC), serum creatinine and blood glucose levels between the two genders were statistically significant. Lipid profile measurements did not differ significantly between genders, but mean and median values of triglycerides (TG) between male and female rats showed a difference of more than 10 mg/dL. The liver enzymes alkaline phosphatase (AP) and aspartate aminotransferase (AST) were also statistically significantly different between sexes. Despite wide variation in mean alanine aminotransferase (ALT) between sexes, the difference was not statistically significant. CONCLUSION: The findings of this project should support to a certain extent the "Reduction" aspect of the 3Rs concept of Russell and Burch by reducing the number of Sprague-Dawley rats used in future research projects at UCFM.

PhenoWorld: addressing animal welfare in a new paradigm to house and assess rat behaviour.

The use of animals is essential in biomedical research. The laboratory environment where the animals are housed has a major impact on them throughout their lives and influences the outcome of animal experiments. Therefore, there has been an increased effort in the refinement of laboratory housing conditions which is explicitly reflected in international regulations and recommendations. Since housing conditions affect behaviour and brain function as well as well-being, the validation of an animal model or paradigm to study the brain and central nervous system disorders is not complete without an evaluation of its implication on animal welfare. Here we discuss several aspects of animal welfare, comparing groups of six rats living in the PhenoWorld (PhW), a recently developed and validated paradigm for studying rodent behaviour, with standard-housed animals (in cages of six rats or pair-housed). In this study we present new data on home-cage behaviour showing that PhW animals have a clearer circadian pattern of sleep and social interaction. We conclude that, by promoting good basic health and functioning, together with the performance of natural behaviours, and maintaining animals' control over some of their environment but still keeping some physical and social challenges, the PhW stimulates positive affective states and higher motivation in rats, which might contribute to an increased welfare for animals living in the PhW.

The effects of different types of individually ventilated caging systems on growing male mice.

Ventilation rate and turnover rate of dry air vary among different types of ventilation systems used with individually ventilated cages (IVCs) and can affect the well-being of rodents housed in these cages. The authors compared the effects of two types of IVC systems, forced-air IVCs and motor-free IVCs, on 4-week-old C57Bl/6J male mice. The mice were acclimatized to the cages for 8 d and then monitored for 87 d. Their body weights, food and water consumption and preferred resting areas were recorded. Mice that were housed in motor-free IVCs had a significantly greater increase in body weight than those housed in forced-air IVCs, despite having similar food consumption. Mice in forced-air IVCs had greater water consumption than mice in motor-free IVCs. In addition, mice in forced-air IVCs were more frequently located in the front halves of their cages, whereas mice in motor-free IVCs were located with similar frequency in the front and back halves of their cages, perhaps because of the higher ventilation rate or the location of the air inlets and outlets in the rear of the cage. These results suggest that body weight, food and water consumption and intracage location of growing male mice are influenced by the type of ventilation system used in the cages in which the mice are housed.

Animal models for arthritis: innovative tools for prevention and treatment.

The development of novel treatments for rheumatoid arthritis (RA) requires the interplay between clinical observations and studies in animal models. Given the complex molecular pathogenesis and highly heterogeneous clinical picture of RA, there is an urgent need to dissect its multifactorial nature and to propose new strategies for preventive, early and curative treatments. Research on animal models has generated new knowledge on RA pathophysiology and aetiology and has provided highly successful paradigms for innovative drug development. Recent focus has shifted towards the discovery of novel biomarkers, with emphasis on presymptomatic and emerging stages of human RA, and towards addressing the pathophysiological mechanisms and subsequent efficacy of interventions that underlie different disease variants. Shifts in the current paradigms underlying RA pathogenesis have also led to increased demand for new (including humanised) animal models. There is therefore an urgent need to integrate the knowledge on human and animal models with the ultimate goal of creating a comprehensive 'pathogenesis map' that will guide alignment of existing and new animal models to the subset of disease they mimic. This requires full and standardised characterisation of all models at the genotypic, phenotypic and biomarker level, exploiting recent technological developments in 'omics' profiling and computational biology as well as state of the art bioimaging. Efficient integration and dissemination of information and resources as well as outreach to the public will be necessary to manage the plethora of data accumulated and to increase community awareness and support for innovative animal model research in rheumatology.

There's a rat in my room! now what? Mice show no chronic physiological response to the presence of rats.

In general, guidelines on housing and care of animals in the laboratory state that rats and mice should not be housed in the same room. Mice may perceive rats as predators. Although one theory says this can cause stress, there is little scientific evidence to support this theory. In the wild, rats and mice usually do not share the same microhabitat, but this appears to be true for most small rodent species. Furthermore, reports of predatory behavior of rats toward mice mainly originate from experimental settings using rats with high inbred levels of aggression. This experiment measured heart rate (HR), body temperature (BT), activity (AC), and urinary corticosterone in female C57BL/6 mice before, during, and after introducing Wistar rats into their room. The study found no chronic effects of rat introduction on any parameters. The study concluded that housing rats and mice in the same room is at least less disturbing than cage cleaning, which caused a temporary increase of HR, BT, and AC. Current results do not support legislation based on compromised welfare.

Impact of 'living apart together' on postoperative recovery of mice compared with social and individual housing.

Social housing is the optimal way of housing female laboratory mice. However, individual housing may be required in experimental designs, for example after surgery. We therefore investigated whether housing two female mice in a cage, separated by a grid partition ('living apart together', LAT), counters the adverse effects of individual housing on postoperative recovery. Ten individually housed (IND) mice, nine socially housed (SOC) mice and nine mice, housed LAT, were surgically implanted with a telemetry transmitter. From one week prior to surgery until three weeks thereafter, several physiological and behavioural parameters were measured in the mice subjected to surgery. The telemetry transmitter measured heart rate (HR), body temperature and activity continuously. Body weight, food and water intake were scored regularly, as were wound healing, ease of handling, nest building and behaviour. Results indicated that SOC mice appear to be less affected by abdominal surgery than IND mice, as indicated by HR and behaviour. LAT, however, did not appear to be beneficiary to the mice. Increased HR levels and differences in behaviour as compared with both SOC and IND animals indicate that LAT may even be the most stressful of the three housing conditions. We therefore conclude that mice benefit most from social housing after surgery. If, however, social housing is not possible, individual housing appears to be a better option than separating mice by a grid partition.

The influence of the location of a nest box in an individually ventilated cage on the preference of mice to use it.

The improvement of housing conditions for mice by using environmental enrichment materials is of high concern for the scientific community. Plastic, autoclavable nest boxes are commercially available and ready to use for specific cases such as in individually ventilated cages, metabolic cages, or during toxicological studies. The aim of this study was to see if the location of the nest box within the cage could influence the mice to prefer and use it. Located on the cage floor or hung from the cage lid, a nest box (MPLEX, Otto Environmental, Milwaukee, Wisconsin), enriched the cages. The study concluded that the location of the nest boxes in the individually ventilated cage plays a significant role in the mice preferring to use it or to avoid it. It is also important to use environmental enrichment items that provide animals with the possibility of expressing their preferences and manipulating them in a way to cope better with their environmental conditions.

Neurotrophin levels and behaviour in BALB/c mice: impact of intermittent exposure to individual housing and wheel running.

This study assessed the effects of intermittent individual housing on behaviour and brain neurotrophins, and whether physical exercise could influence alternate individual-housing-induced effects. Five-week-old BALB/c mice were either housed in enhanced social (E) or standard social (S) housing conditions for 2 weeks. Thereafter they were divided into six groups and for 6 weeks remained in the following experimental conditions: Control groups remained in their respective housing conditions (E-control, S-control); enhanced individual (E-individual) and standard individual (S-individual) groups were exposed every other day to individual cages without running-wheels; enhanced running-wheel (E-wheel) and standard running-wheel (S-wheel) groups were put on alternate days in individual running-wheel cages. Animals were assessed for activity in an automated individual cage system (LABORAS) and brain neurotrophins analysed. Intermittent individual housing increased behavioural activity and reduced nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) levels in frontal cortex; while it increased BDNF level in the amygdala and BDNF protein and mRNA in hippocampus. Besides normalizing motor activity and regulating BDNF and NGF levels in hippocampus, amygdala and cerebellum, physical exercise did not attenuate reduction of cortical NGF and BDNF induced by intermittent individual housing. This study demonstrates that alternate individual housing has significant impact on behaviour and brain neurotrophin levels in mice, which can be partially altered by voluntary physical exercise. Our results also suggest that some changes in neurotrophin levels induced by intermittent individual housing are not similar to those caused by continuous individual housing.

Intermittent individual housing increases survival of newly proliferated cells.

In this study, we analyzed how intermittent individual housing with or without a running wheel influenced corticosterone levels and survival of newly proliferated cells in the dentate gyrus of the hippocampus. Female Balb/c mice, in standard or enhanced housing, were divided into groups that were individually housed with or without running wheels on every second day. Intermittent individual housing without, but not with, running wheels increased survival of proliferated cells in the dentate gyrus as compared with continuous group housing in standard or enhanced conditions. Thus, changes in housing conditions on every second day can, under certain circumstances, have an impact on the survival of newly proliferated cells in the dentate gyrus.

Environmental enrichment for laboratory rodents and rabbits: requirements of rodents, rabbits, and research.

Environmental conditions such as housing and husbandry have a major impact on the laboratory animal throughout its life and will thereby influence the outcome of animal experiments. However, housing systems for laboratory animals have often been designed on the basis of economic and ergonomic aspects. One possible way to improve the living conditions of laboratory animals is to provide opportunities for the animals to perform a species-specific behavioral repertoire. Environmental enrichment should be regarded both as an essential component of the overall animal care program and equally important as nutrition and veterinary care. The key component of an enrichment program is the animal staff, whose members must be motivated and educated. It is critically important to evaluate environmental enrichment in terms of the benefit to the animal by assessing the use of and preference for a certain enrichment, the effect on behavior and the performance of species-typical behavior, and the effect on physiological parameters. At the same time, it is necessary to evaluate the impact on scientific outcome, how the enrichment influences the scientific study, and whether and how the statistical power is affected. The result will depend on the parameter measured, the type of enrichment used, and the animal strain. In this article, goals of enrichment are defined and discussed. Animal behaviors and needs are described, along with the translation of those needs into environmental enrichment programs. Specific types of environmental enrichment are outlined with examples from the literature, and an evaluation of environmental enrichment is provided.

Assessment of the use of two commercially available environmental enrichments by laboratory mice by preference testing.

In the field of biomedical research, the demand for standardization of environmental enrichment for laboratory animals is growing. For laboratory mice, a wide variety of environmental enrichment items are commercially available. Most of these comply with the demands for standardization, hygiene and ergonomics. Whether these items also comply with their actual purpose, to enhance the well-being of the mice, is often not assessed scientifically. In this study, we tested the preference of mice for two commercially available nest boxes differing in shape and material: the Shepherd Shack/DesRes (SS/DR) and the Tecniplast Mouse House (TMH), in a simple preference test. To indicate strength of preference, both nest boxes were also tested against a highly preferred nesting material. Preference for the most preferred nest box was investigated further. Our results indicated a strong preference by mice for the SS/DR, but not for the TMH. Furthermore, nesting material was almost always combined with the SS/DR, but not with the TMH. More elaborate testing of the SS/DR in an automated preference test system confirmed that mice spent significantly more time in a cage in which an SS/DR is provided. Differences between both nest boxes are discussed with regard to their attractiveness to mice. It is also argued that enrichment should primarily be developed in concordance with the animals' needs prior to the marketing of enrichment tools.

Urinary corticosterone levels in mice in response to intraperitoneal injections with saline.

The concept of refinement is an important issue in the field of laboratory animal science. Refinement-based research aims to improve animal welfare, to increase the reliability of experimental outcome, and to diminish variation. In search of refinement of experimental techniques, this study investigated whether urinary corticosterone can be used as a noninvasive measure of acute stress in mice.

Behavioral assessment of intermittent wheel running and individual housing in mice in the laboratory.

Physical cage enrichment--exercise devices for rodents in the laboratory--often includes running wheels. This study compared responses of mice in enriched physical and social conditions and in standard social conditions to wheel running, individual housing, and open-field test. The study divided into 6 groups, 48 female BALB/c mice group housed in enriched and standard conditions. On alternate days, the study exposed 2 groups to individual running wheel cages. It intermittently separated from their cage mates and housed individually 2 groups with no running wheels; 2 control groups remained in enriched or standard condition cages. There were no significant differences between enriched and standard group housed mice in alternate days' wheel running. Over time, enriched, group housed mice ran less. Both groups responded similarly to individual housing. In open-field test, mice exposed to individual housing without running wheel moved more and faster than wheel running and home cage control mice. They have lower body weights than group housed and wheel running mice. Intermittent withdrawal of individual housing affects the animals more than other commodities. Wheel running normalizes some effects of intermittent separation from the enriched, social home cage.

Environmental enrichment: room for reduction?

Environmental enrichment strategies are usually regarded as refinement. However, when the welfare of animals is enhanced through successful enrichment programmes, a reduction in the number of animals needed can be expected, because fewer animals might be lost during the course of experiments. Several examples of studies where enrichment can lead to reduction will be presented. They include the beneficial effects of nesting material for laboratory mice, the effects of husbandry procedures on controlling aggressive behaviour in male laboratory mice, and the effects of enrichment on variation in the results of experiments.