Research

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14 pages, 2641 KiB

Open AccessArticle

Impact of Refinements to Handling and Restraint Methods in Mice

by Jennifer R. Davies, Dandri A. Purawijaya, Julia M. Bartlett and Emma S. J. Robinson

Animals 2022, 12(17), 2173; https://doi.org/10.3390/ani12172173 - 24 Aug 2022

Cited by 6 | Viewed by 2916 | Correction

There is increasing evidence that, compared to non-aversive handling methods (i.e., tunnel and cupping), tail handling has a negative impact on mouse welfare. Despite this evidence, there are still research organisations that continue to use tail handling. Here, we investigated handling for routine [...] Read more.

There is increasing evidence that, compared to non-aversive handling methods (i.e., tunnel and cupping), tail handling has a negative impact on mouse welfare. Despite this evidence, there are still research organisations that continue to use tail handling. Here, we investigated handling for routine husbandry by three different methods: tail, cupping and tube in a relevant real-world scenario involving mice bred off-site. After transfer to the destination unit, mice were assessed for overt behaviours associated with anxiety and fear. Mice that experienced tail handling were less easy to handle, were more responsive to the box opening, and scored lower in a hand approach test. One barrier to non-tail handling methods is the current practice of restraining mice by the tail for procedures. We therefore next assessed whether a modified method for restraint that takes the animal from cupping to restraint without the use of the tail was associated with better welfare. This refined restraint method reduced overt signs of distress although we did not find any differences in corticosterone levels or anxiety-related behaviours. These findings suggest that avoiding tail handling throughout the animal’s laboratory experience, including during restraint, benefits their welfare. Full article

(This article belongs to the Special Issue Improving Research Animal Welfare and Quality of Science)

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19 pages, 16717 KiB

Open AccessArticle

A Refined Method for Studying Foraging Behaviour and Body Mass in Group-Housed European Starlings

by Melissa Bateson and Ryan Nolan

Animals 2022, 12(9), 1159; https://doi.org/10.3390/ani12091159 - 29 Apr 2022

Cited by 3 | Viewed by 1770

Abstract

Laboratory experiments on passerine birds have been important for testing hypotheses regarding the effects of environmental variables on the adaptive regulation of body mass. However, previous work in this area has suffered from poor ecological validity and animal welfare due to the requirement [...] Read more.

Laboratory experiments on passerine birds have been important for testing hypotheses regarding the effects of environmental variables on the adaptive regulation of body mass. However, previous work in this area has suffered from poor ecological validity and animal welfare due to the requirement to house birds individually in small cages to facilitate behavioural measurement and frequent catching for weighing. Here, we describe the social foraging system, a novel technology that permits continuous collection of individual-level data on operant foraging behaviour and body mass from group-housed European starlings (Sturnus vulgaris). We report on the rapid acquisition of operant key pecking, followed by foraging and body mass data from two groups of six birds maintained on a fixed-ratio operant schedule under closed economy for 11 consecutive days. Birds gained 6.0 ± 1.2 g (mean ± sd) between dawn and dusk each day and lost an equal amount overnight. Individual daily mass gain trajectories were non-linear, with the rate of gain decelerating between dawn and dusk. Within-bird variation in daily foraging effort (key pecks) positively predicted within-bird variation in dusk mass. However, between-bird variation in mean foraging effort was uncorrelated with between-bird variation in mean mass, potentially indicative of individual differences in daily energy requirements. We conclude that the social foraging system delivers refined data collection and offers potential for improving our understanding of mass regulation in starlings and other species. Full article

(This article belongs to the Special Issue Improving Research Animal Welfare and Quality of Science)

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19 pages, 2339 KiB

Open AccessArticle

Welfare Assessment, End-Point Refinement and the Effects of Non-Aversive Handling in C57BL/6 Mice with Lewis Lung Cancer

by Amy L. Miller and Johnny V. Roughan

Animals 2022, 12(1), 23; https://doi.org/10.3390/ani12010023 - 23 Dec 2021

Cited by 3 | Viewed by 3105

Abstract

Cancer-bearing mice are at risk of developing anxiety, pain, or malaise. These conditions may not only harm welfare but could also undermine data quality and translational validity in studies to develop therapeutic interventions. We aimed to establish whether, or at what point mice [...] Read more.

Cancer-bearing mice are at risk of developing anxiety, pain, or malaise. These conditions may not only harm welfare but could also undermine data quality and translational validity in studies to develop therapeutic interventions. We aimed to establish whether, or at what point mice developing lung cancer show these symptoms, what measures can best detect their onset, and if data quality and animal welfare can be enhanced by using non-aversive handling (NAH). Welfare was monitored using various daily methods. At the beginning and end of the study, we also scored behaviour for general welfare evaluation, recorded nociceptive thresholds, and applied the mouse grimace scale (MGS). Cancer caused a decline in daily welfare parameters (body weight, and food and water consumption) beginning at around 4 days prior to euthanasia. As cancer progressed, rearing and walking declined to a greater extent in cancer-bearing versus control mice, while grooming, inactive periods, and MGS scores increased. A decline in nest building capability and food consumption provided a particularly effective means of detecting deteriorating welfare. These changes suggested a welfare problem arose as cancer developed, so similar studies would benefit from refinement, with mice being removed from the study at least 4 days earlier. However, the problem of highly varied tumour growth made it difficult to determine this time-point accurately. There were no detectable beneficial effects of NAH on either data quality or in terms of enhanced welfare. Full article

(This article belongs to the Special Issue Improving Research Animal Welfare and Quality of Science)

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18 pages, 793 KiB

Open AccessArticle

Temperament Predicts the Quality of Social Interactions in Captive Female Rhesus Macaques (Macaca mulatta)

by Ori Pomerantz and John P. Capitanio

Animals 2021, 11(8), 2452; https://doi.org/10.3390/ani11082452 - 20 Aug 2021

Cited by 4 | Viewed by 2763

Abstract

Previous reports suggest that female macaques with greater similarity in emotionality and nervous temperament, as evaluated in a well-established BioBehavioral Assessment (BBA) at the California National Primate Research Center, were more likely to form successful pairs. We tested whether the same measures can [...] Read more.

Previous reports suggest that female macaques with greater similarity in emotionality and nervous temperament, as evaluated in a well-established BioBehavioral Assessment (BBA) at the California National Primate Research Center, were more likely to form successful pairs. We tested whether the same measures can also predict the quality of social interactions among 20 female rhesus macaque pairs. We correlated the pairs’ emotionality and nervous temperament scores obtained in infancy and the levels of behaviors recorded systematically during the pairing process years later. Supporting previous findings, partners with similar emotionality scores were more affiliative, and pairs with similar nervous temperament expressed less dominance/submissive behavior. Exploratorily, we found that pairs that were better at processing social information (part of BBA) were also more anxious. Such animals should be prioritized to be introduced in rooms that house calmer, less aggressive animals and provide opportunities for hiding to alleviate their anxiety. Indeed, positive social experiences not only promote animal welfare, but also reduce stress related confounds and unexplained data variability. Therefore, by incorporating the animals’ temperament into the pair configuration process we increase the likelihood of forming high-quality pairs, both in terms of welfare and the research of which they are a part. Full article

(This article belongs to the Special Issue Improving Research Animal Welfare and Quality of Science)

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12 pages, 774 KiB

Open AccessArticle

Improved Training and Semen Collection Outcomes Using the Closed Box Chair for Macaques

by Lisa A. Houser, Cathy Ramsey, Fernanda M. de Carvalho, Breanna Kolwitz, Chelsey Naito, Kristine Coleman and Carol B. Hanna

Animals 2021, 11(8), 2384; https://doi.org/10.3390/ani11082384 - 12 Aug 2021

Cited by 5 | Viewed by 3080

Abstract

Collaborative semen collection in monkeys is a valuable tool in research, animal collection management, and conservation efforts. To obtain samples, monkeys are often restrained in open restraint chairs (ORC) with the “pole and collar” technique. While commonly used, this restraint is not tolerated [...] Read more.

Collaborative semen collection in monkeys is a valuable tool in research, animal collection management, and conservation efforts. To obtain samples, monkeys are often restrained in open restraint chairs (ORC) with the “pole and collar” technique. While commonly used, this restraint is not tolerated by all individuals; some become anxious or aggressive towards the poles and people. In an effort to refine this procedure and improve welfare of the monkeys, we examined the use of a “closed box chair” (CBC), a clear, plexiglass box in which the monkey is trained to sit for sperm collection. The CBC does not require pole and collar, and although legs are secured, the arms and neck are not restrained. The use of CBCs has increased in recent years; however, there are few studies demonstrating its effects on scientific outcomes. We used positive reinforcement techniques to train 34 adult male rhesus macaques (Macaca mulatta) to provide semen samples using either the ORC or the CBC. While all CBC monkeys (n = 14) were reliably trained for this procedure, only 75% of ORC (n = 20) males completed the training (p = 0.04). It took significantly less time to train animals in the CBC than the ORC (201.0 vs. 412.4 min; p <0.001). In a controlled subset, males restrained with ORC (n = 7) produced a significantly lower ejaculatory volume than those collected by CBC (n = 10) (297.6 µL vs. 522.1 µL respectively; p = 0.04) and had a lower concentration of sperm (186.0 × 10⁶/mL vs. 367.5 × 10⁶/mL respectively; p = 0.017), although there were no differences with respect to sperm motility (p = 0.15). Our data suggest the closed box chair technique reduces stress on the animals while enhancing semen quality, supporting the use of the CBC as an important refinement. Full article

(This article belongs to the Special Issue Improving Research Animal Welfare and Quality of Science)

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Review

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15 pages, 324 KiB

Open AccessReview

Using Mice to Model Human Disease: Understanding the Roles of Baseline Housing-Induced and Experimentally Imposed Stresses in Animal Welfare and Experimental Reproducibility

by Bonnie L. Hylander, Elizabeth A. Repasky and Sandra Sexton

Animals 2022, 12(3), 371; https://doi.org/10.3390/ani12030371 - 03 Feb 2022

Cited by 4 | Viewed by 3698

Abstract

Mice are the most common animal used to study disease, but there are real concerns about the reproducibility of many of these experiments. This review discusses how several different sources of chronic stress can directly impact experimental outcomes. Mandated housing conditions induce an [...] Read more.

Mice are the most common animal used to study disease, but there are real concerns about the reproducibility of many of these experiments. This review discusses how several different sources of chronic stress can directly impact experimental outcomes. Mandated housing conditions induce an underappreciated level of chronic stress but are not usually considered or reported as part of the experimental design. Since chronic stress plays a critical role in the development and progression of many somatic diseases including cancer, obesity, and auto-immune diseases, this baseline stress can directly affect outcomes of such experiments. To study the role of stress in both physical and psychiatric diseases, there has been a proliferation of protocols for imposing chronic stress on mice. For somatic diseases, biomarkers can be used to compare the models with the disease in patients, but to evaluate the validity of psychiatric models, behavioral tests are carried out to assess changes in behavior and these tests may themselves cause an underappreciated degree of additional stress. Therefore, it is important for animal welfare to reduce baseline stress and select the most humane protocols for inducing and assessing chronic stress to obtain the most reliable outcomes. Full article

(This article belongs to the Special Issue Improving Research Animal Welfare and Quality of Science)

16 pages, 25838 KiB

Open AccessReview

Refining Procedures within Regulatory Toxicology Studies: Improving Animal Welfare and Data

by Helen Prior, Hollie Blunt, Lee Crossman, Aidan McGuire, Ruth Stow and Fiona Sewell

Animals 2021, 11(11), 3057; https://doi.org/10.3390/ani11113057 - 26 Oct 2021

Cited by 3 | Viewed by 2774

Abstract

During the development of potential new medicines or agrochemicals, an assessment of the safety profile to humans and environmental species is conducted using a range of different in silico and in vitro techniques in conjunction with metabolism and toxicity studies using animals. The [...] Read more.

During the development of potential new medicines or agrochemicals, an assessment of the safety profile to humans and environmental species is conducted using a range of different in silico and in vitro techniques in conjunction with metabolism and toxicity studies using animals. The required studies are outlined within international regulatory guidelines which acknowledge and support the application of the 3Rs to reduce the number of animals used or to refine the procedures performed when these studies are deemed to be necessary. The continued development of new technologies and adoption of best-practice approaches to laboratory animal housing and study procedures has generated a series of refinements that can be incorporated into animal studies throughout the package. These refinements benefit the welfare of fish, mice, rats, rabbits, dogs, minipigs, and non-human primates (NHPs) whilst maintaining or improving data quality within general toxicology, metabolism, and other studies and can also bring efficiencies to processes that benefit study costs and timings. Examples are shared which cover the following topics: social housing of dogs and NHPs, surgical refinements in the rat bile duct cannulation model for collection of data for metabolism studies, whether fasting is really required prior to clinical pathology sampling, and the use of microsampling for toxicokinetics. Full article

(This article belongs to the Special Issue Improving Research Animal Welfare and Quality of Science)

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21 pages, 1103 KiB

Open AccessReview

Knowledge of Biobehavioral Organization Can Facilitate Better Science: A Review of the BioBehavioral Assessment Program at the California National Primate Research Center

by John P. Capitanio

Animals 2021, 11(8), 2445; https://doi.org/10.3390/ani11082445 - 20 Aug 2021

Cited by 11 | Viewed by 2848

Abstract

Animals vary on intrinsic characteristics such as temperament and stress responsiveness, and this information can be useful to experimentalists for identifying more homogeneous subsets of animals that show consistency in risk for a particular research outcome. Such information can also be useful for [...] Read more.

Animals vary on intrinsic characteristics such as temperament and stress responsiveness, and this information can be useful to experimentalists for identifying more homogeneous subsets of animals that show consistency in risk for a particular research outcome. Such information can also be useful for balancing experimental groups, ensuring animals within an experiment have similar characteristics. In this review, we describe the BioBehavioral Assessment Program at the California National Primate Research Center, which, since its inception in 2001, has been providing quantitative information on intrinsic characteristics to scientists for subject selection and balancing, and to colony management staff for management purposes. We describe the program and review studies relating to asthma, autism, behavioral inhibition, etc., where the BBA Program was used to select animals. We also review our work, showing that factors such as rearing, ketamine exposure, and prenatal experience can affect biobehavioral organization in ways that some investigators might want to control for in their studies. Attention to intrinsic characteristics of subject populations is consistent with the growing interest in precision medicine and can lead to a reduction in animal numbers, savings in time and money for investigators, and reduced distress for the animals. Full article

(This article belongs to the Special Issue Improving Research Animal Welfare and Quality of Science)

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