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A special issue of Animals (ISSN 2076-2615). This special issue belongs to the section "Veterinary Clinical Studies".

Deadline for manuscript submissions: closed (31 July 2020) | Viewed by 72132

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Dr. Gabrielle C. Musk

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Animal Care Services, the University of Western Australia, Perth, Australia
Interests: analgesia; pain; production animals; cattle; sheep; pigs; poultry
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

The value of publications focused specifically on refinement of animal models in biomedical research is immense. Relying on authors to report their science and include adequate details on methodology, adverse events and outcomes for animals used in biomedical research is unlikely to uphold the 3Rs (refine, reduce, replace) of animal research. Despite the aims of the ARRIVE (Animals in Research: Reporting in vivo Experiments) guidelines published in 2010 and revised in 2019, there is still room for improvement in the level of detail included in publications ^{1, 2}. As examples, details on husbandry, anaesthesia, analgesia, post-procedural monitoring, morbidity and mortality are often scant and therefore make efforts at refinement of models more challenging.

Refinement refers to optimizing the welfare of research animals by avoiding or minimising pain, fear and distress and by maintaining animals in conditions that promote their health and well-being ³. Key contributors to efforts at refinement include anaesthesia, analgesia and post-operative care of animals. Ultimately, refinements should achieve improvements in animal welfare and quality of science.

We invite submissions describing efforts at refinement to animal models to increase the evidence base and to identify and explore opportunities for improving animal welfare and the quality of science.

Kilkenny C, Browne WJ, Cuthill IC, et al. Improving Bioscience Research Reporting: The ARRIVE Guidelines for Reporting Animal Research. PLOS Biology 2010; 8: e1000412. DOI: 10.1371/journal.pbio.1000412.
Percie du Sert N, Hurst V, Ahluwalia A, et al. The ARRIVE guidelines 2019: updated guidelines for reporting animal research. bioRxiv 2019: 703181. DOI: 10.1101/703181.
Prescott MJ and Lidster K. Improving quality of science through better animal welfare: the NC3Rs strategy. Lab Anim (NY) 2017; 46: 152-156. 2017/03/23. DOI: 10.1038/laban.1217.

Dr. Gabrielle Musk
Guest Editor

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Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

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Keywords

3Rs
animal model
refinement
animal welfare

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Published Papers (13 papers)

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Editorial

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3 pages, 159 KiB

Open AccessEditorial

Refinements to Animal Models for Biomedical Research

by Gabrielle C. Musk

Animals 2020, 10(12), 2425; https://doi.org/10.3390/ani10122425 - 18 Dec 2020

Cited by 4 | Viewed by 2205

Abstract

This collection includes the manuscripts published in the Special Issue of Animals: Refinements to Animal Models for Biomedical Research [...] Full article

(This article belongs to the Special Issue Refinements to Animal Models for Biomedical Research)

Research

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12 pages, 2146 KiB

Open AccessArticle

Fentanyl Plasma Concentrations after Application of a Transdermal Patch in Three Different Locations to Refine Postoperative Pain Management in Rabbits

by Valentina Mirschberger, Christian von Deimling, Anja Heider, Claudia Spadavecchia, Helene Rohrbach and Stephan Zeiter

Animals 2020, 10(10), 1778; https://doi.org/10.3390/ani10101778 - 1 Oct 2020

Cited by 9 | Viewed by 3744

Abstract

Transdermal patches allow a noninvasive and “stress free” analgesia in rabbits. As fentanyl uptake is dependent on exogenous and endogenous factors of the area where the patch is applied, this study investigated three different locations (neck, inner and outer surfaces of the ear) for fentanyl patch application to provide adequate and reliable fentanyl plasma concentrations above those previously shown to be analgesic. Fentanyl plasma concentration was measured at different time points (3, 6, 9, 12, 18, 24, 36, 48, 72, 96, 120 h) and rabbits were assessed for their general conditions and treatment-related side effects. Practicability of the proposed methods was evaluated. Following patch application on the neck, fentanyl plasma concentrations equal to or above the analgesic value were measured in all rabbits between 6 and 72 h. Comparable concentrations were reached between 9 and 48 h in all animals for the outer ear surface. However, for the inner ear surface, analgesic concentrations were not reached, even if practicability was considered the best for this location. Preparation of the neck skin was judged as the most cumbersome due to the clipping of the dense fur and patch removal resulted in erythema. In summary, the application of the fentanyl patch on the neck and outer ear surface allowed the reach of reliable plasma concentrations above the analgesic threshold in rabbits. When applied on the neck, fentanyl patches provided the longest duration of analgesic plasma concentrations, whereas patch application and removal were easier on the outer ear surface. Full article

(This article belongs to the Special Issue Refinements to Animal Models for Biomedical Research)

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14 pages, 4332 KiB

Open AccessFeature PaperArticle

Transdermal Fentanyl Uptake at Two Different Patch Locations in Swiss White Alpine Sheep

by Tim Buchholz, Maria Hildebrand, Anja Heider, Valentina Stenger, Daniel Arens, Claudia Spadavecchia and Stephan Zeiter

Animals 2020, 10(9), 1675; https://doi.org/10.3390/ani10091675 - 17 Sep 2020

Cited by 8 | Viewed by 8471

Abstract

When using animals in biomedical research, investigators have the responsibility to ensure adequate analgesia. Currently, transdermal fentanyl patches (TFP) are often used to provide postoperative analgesia in large laboratory animals. The aim of this study was to compare the fentanyl uptake resulting from TFP applied at two different locations, namely the foreleg and the thorax, in healthy adult sheep. Twelve sheep received a TFP with an intended dosage of 2 ug/kg/h. Blood samples were taken at different time points over a period of five days and the fentanyl plasma levels were measured. The TFP applied on the foreleg allowed a faster fentanyl uptake with higher peaks and a longer time within or above the target concentration of 0.6–1.5 ng/mL, shown to be analgesic in humans, when compared to the one on the thorax. Assuming that the effective plasma concentration described for humans is providing analgesia in sheep as well, the present findings suggest that it should be sufficient to apply the TFP 3–6 h before the painful insult and that its effect should last at least 48 h. Furthermore, when TFP are used to provide postoperative analgesia in sheep, they should be placed on the foreleg rather than on the thorax. Full article

(This article belongs to the Special Issue Refinements to Animal Models for Biomedical Research)

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20 pages, 3798 KiB

Open AccessArticle

Remote Controlled Nociceptive Threshold Testing Systems in Large Animals

by Polly Taylor

Animals 2020, 10(9), 1556; https://doi.org/10.3390/ani10091556 - 2 Sep 2020

Cited by 9 | Viewed by 3259

Abstract

Nociceptive threshold (NT) testing is widely used for the study of pain and its alleviation. The end point is a normal behavioural response, which may be affected by restraint or unfamiliar surroundings, leading to erroneous data. Remotely controlled thermal and mechanical NT testing systems were developed to allow free movement during testing and were evaluated in cats, dogs, sheep, horses and camels. Thermal threshold (TT) testing incorporated a heater and temperature sensor held against the animal’s shaved skin. Mechanical threshold (MT) testing incorporated a pneumatic actuator attached to a limb containing a 1–2 mm radiused pin pushed against the skin. Both stimuli were driven from battery powered control units attached on the animal’s back, controlled remotely via infra-red radiation from a handheld component. Threshold reading was held automatically and displayed digitally on the unit. The system was failsafe with a safety cut-out at a preset temperature or force as appropriate. The animals accepted the equipment and behaved normally in their home environment, enabling recording of reproducible TT (38.5–49.8 °C) and MT (2.7–10.1 N); precise values depended on the species, the individual and the stimulus characteristics. Remote controlled NT threshold testing appears to be a viable refinement for pain research. Full article

(This article belongs to the Special Issue Refinements to Animal Models for Biomedical Research)

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5 pages, 487 KiB

Open AccessCommunication

Survey on Sheep Usage in Biomedical Research

by Corina Mihaela Berset, Urban Lanker and Stephan Zeiter

Animals 2020, 10(9), 1528; https://doi.org/10.3390/ani10091528 - 30 Aug 2020

Cited by 8 | Viewed by 2587

Abstract

Currently, there is a lack of detailed information about sheep used for biomedical research. Therefore, a European survey was conducted among sheep users gathering information on the current situation, with emphasis on animal selection criteria and issues encountered in practice. The ultimate goal was to identify needs for improvement, which will subsequently lead to a refinement and reduction of the total number of animals used for experimental studies. From the 84 respondents, 77.4% were veterinarians, 71.4% were employed at academic institutions and 63.1% had worked with sheep as research animals for more than 5 years. The majority of the respondents were using females (79.8%) with no clear age preference, mainly for surgical procedures and testing medical devices. The main criteria for choosing a sheep supplier were the animals’ health status, their availability, the trust and experience in the sheep provider and the animals’ uniformity. Approximately 60% of the respondents had encountered problems in their sheep not related to the experimental protocol and almost half of them did not have a health monitoring program for their animals. In conclusion, there is definitely a need for refinement in selecting sheep used in biomedical research, with their health status as possible starting point. Full article

(This article belongs to the Special Issue Refinements to Animal Models for Biomedical Research)

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14 pages, 1287 KiB

Open AccessArticle

Humane Euthanasia of Guinea Pigs (Cavia porcellus) with a Penetrating Spring-Loaded Captive Bolt

by Shari Cohen, Melody Kwok and Joel Huang

Animals 2020, 10(8), 1356; https://doi.org/10.3390/ani10081356 - 5 Aug 2020

Cited by 4 | Viewed by 9170

Abstract

Guinea pigs (Cavia porcellus) have been used in research since the 19th century to collect data due to their physiological similarities to humans. Today, animals perform a vital role in experiments and concerns for laboratory animal welfare are enshrined in the 3R framework of reduction, refinement and replacement. This case study explores a refinement in humane euthanasia of guinea pigs via the use of an irreversible penetrating spring-loaded captive bolt (CB). Penetrating spring-loaded CB stunning for euthanasia (CBE) was performed on 12 guinea pigs with the parameters for humane slaughter of production animals in order to assess the suitability of this method of euthanasia in contrast to blunt force trauma (BFT). All 12 of the guinea pigs were rendered immediately unconscious with excellent experimental tissue quality collection, high repeatability of results and operator (n = 8) preference over BFT. Overall, CBE in guinea pigs appears to be a feasible refinement for animal welfare, human preference and improved tissue quality for experimental collection in settings where uncontaminated tissues are required. Full article

(This article belongs to the Special Issue Refinements to Animal Models for Biomedical Research)

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14 pages, 573 KiB

Open AccessArticle

Differential Transcription of Selected Cytokine and Neuroactive Ligand-receptor Genes in Peripheral Leukocytes from Calves in Response to Cautery Disbudding

by Kavitha Kongara, Venkata Sayoji Rao Dukkipati, Hui Min Tai, Axel Heiser, Alan Murray, James Webster and Craig Brian Johnson

Animals 2020, 10(7), 1187; https://doi.org/10.3390/ani10071187 - 14 Jul 2020

Cited by 6 | Viewed by 3182

Abstract

Calf disbudding is a painful husbandry practice on dairy and beef cattle farms. An objective measurement of pain is useful to reliably evaluate the pain intensity and anti-nociceptive (analgesic) efficacy of therapeutic agents. The aim of this study was to investigate the changes in peripheral leucocyte inflammatory cytokine gene expression in calves after disbudding, and to assess whether the changes in cytokine gene expression could be an indicator of the efficacy of analgesic drugs. In a randomised controlled study, 16 calves (aged 31 to 41 days and weighing 58 to 73 kg), undergoing routine disbudding, were randomly allocated into two groups (n = 8 in each group). Calves in the control group received no analgesic, while those in the treatment group received 0.5 mg kg⁻¹ meloxicam subcutaneously prior to disbudding. Disbudding was performed using an electric debudder. Blood (10 mL) was sampled from the jugular vein just before and 4 and 24 h post-disbudding, RNA was extracted from leukocytes, and the transcription of 12 genes of interest was assessed using nCounter gene expression assay. The results showed significantly higher transcription (compared to baseline values) of the studied genes (except CRH, IFNγ, and IL10) in the control group calves at either 4 or 24 h post-disbudding. The administration of meloxicam one hour before disbudding significantly attenuated the upregulation of IL6, PGHS2, TAC1, NOS1, and CRH gene transcription post-disbudding, while it did not suppress the elevated transcription of acute and pro-inflammatory cytokines such as IL1β, IFNγ, IL8, and TNFα genes. In conclusion, nCounter gene expression assay seems to be a promising tool to study the expression of cytokine genes and thus could be used for the pre-clinical evaluation of novel analgesics. Full article

(This article belongs to the Special Issue Refinements to Animal Models for Biomedical Research)

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14 pages, 826 KiB

Open AccessArticle

The Pharmacokinetics of Medetomidine Administered Subcutaneously during Isoflurane Anaesthesia in Sprague-Dawley Rats

by Leila T. Kint, Bhedita J. Seewoo, Timothy H. Hyndman, Michael W. Clarke, Scott H. Edwards, Jennifer Rodger, Kirk W. Feindel and Gabrielle C. Musk

Animals 2020, 10(6), 1050; https://doi.org/10.3390/ani10061050 - 18 Jun 2020

Cited by 14 | Viewed by 4687

Abstract

Anaesthetic protocols involving the combined use of a sedative agent, medetomidine, and an anaesthetic agent, isoflurane, are increasingly being used in functional magnetic resonance imaging (fMRI) studies of the rodent brain. Despite the popularity of this combination, a standardised protocol for the combined use of medetomidine and isoflurane has not been established, resulting in inconsistencies in the reported use of these drugs. This study investigated the pharmacokinetic detail required to standardise the use of medetomidine and isoflurane in rat brain fMRI studies. Using mass spectrometry, serum concentrations of medetomidine were determined in Sprague-Dawley rats during medetomidine and isoflurane anaesthesia. The serum concentration of medetomidine for administration with 0.5% (vapouriser setting) isoflurane was found to be 14.4 ng/mL (±3.0 ng/mL). The data suggests that a steady state serum concentration of medetomidine when administered with 0.5% (vapouriser setting) isoflurane can be achieved with an initial subcutaneous (SC) dose of 0.12 mg/kg of medetomidine followed by a 0.08 mg/kg/h SC infusion of medetomidine. Consideration of these results for future studies will facilitate standardisation of medetomidine and isoflurane anaesthetic protocols during fMRI data acquisition. Full article

(This article belongs to the Special Issue Refinements to Animal Models for Biomedical Research)

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Review

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17 pages, 344 KiB

Open AccessReview

The Utility of Grimace Scales for Practical Pain Assessment in Laboratory Animals

by Daniel Mota-Rojas, Adriana Olmos-Hernández, Antonio Verduzco-Mendoza, Elein Hernández, Julio Martínez-Burnes and Alexandra L. Whittaker

Animals 2020, 10(10), 1838; https://doi.org/10.3390/ani10101838 - 9 Oct 2020

Cited by 41 | Viewed by 8071

Abstract

Animals’ facial expressions are widely used as a readout for emotion. Scientific interest in the facial expressions of laboratory animals has centered primarily on negative experiences, such as pain, experienced as a result of scientific research procedures. Recent attempts to standardize evaluation of facial expressions associated with pain in laboratory animals has culminated in the development of “grimace scales”. The prevention or relief of pain in laboratory animals is a fundamental requirement for in vivo research to satisfy community expectations. However, to date it appears that the grimace scales have not seen widespread implementation as clinical pain assessment techniques in biomedical research. In this review, we discuss some of the barriers to implementation of the scales in clinical laboratory animal medicine, progress made in automation of collection, and suggest avenues for future research. Full article

(This article belongs to the Special Issue Refinements to Animal Models for Biomedical Research)

20 pages, 294 KiB

Open AccessReview

Grimace Scores: Tools to Support the Identification of Pain in Mammals Used in Research

by Shari Cohen and Thierry Beths

Animals 2020, 10(10), 1726; https://doi.org/10.3390/ani10101726 - 23 Sep 2020

Cited by 27 | Viewed by 7081

Abstract

The 3Rs, Replacement, Reduction and Refinement, is a framework to ensure the ethical and justified use of animals in research. The implementation of refinements is required to alleviate and minimise the pain and suffering of animals in research. Public acceptability of animal use in research is contingent on satisfying ethical and legal obligations to provide pain relief along with humane endpoints. To fulfil this obligation, staff, researchers, veterinarians, and technicians must rapidly, accurately, efficiently and consistently identify, assess and act on signs of pain. This ability is paramount to uphold animal welfare, prevent undue suffering and mitigate possible negative impacts on research. Identification of pain may be based on indicators such as physiological, behavioural, or physical ones. Each has been used to develop different pain scoring systems with potential benefits and limitations in identifying and assessing pain. Grimace scores are a promising adjunctive behavioural technique in some mammalian species to identify and assess pain in research animals. The use of this method can be beneficial to animal welfare and research outcomes by identifying animals that may require alleviation of pain or humane intervention. This paper highlights the benefits, caveats, and potential applications of grimace scales. Full article

(This article belongs to the Special Issue Refinements to Animal Models for Biomedical Research)

37 pages, 464 KiB

Open AccessReview

Optimal Methods of Documenting Analgesic Efficacy in Neonatal Piglets Undergoing Castration

by Meredith Sheil and Adam Polkinghorne

Animals 2020, 10(9), 1450; https://doi.org/10.3390/ani10091450 - 19 Aug 2020

Cited by 19 | Viewed by 3312

Abstract

Analgesic products for piglet castration are critically needed. This requires extensive animal experimentation such as to meet regulatory-required proof of efficacy. At present, there are no validated methods of assessing pain in neonatal piglets. This poses challenges for investigators to optimize trial design and to meet ethical obligations to minimize the number of animals needed. Pain in neonatal piglets may be subtle, transient, and/or variably expressed and, in the absence of validated methods, investigators must rely on using a range of biochemical, physiological and behavioural variables, many of which appear to have very low (or unknown) sensitivity or specificity for documenting pain, or pain-relieving effects. A previous systematic review of this subject was hampered by the high degree of variability in the literature base both in terms of methods used to assess pain and pain mitigation, as well as in outcomes reported. In this setting we provide a narrative review to assist in determining the optimal methods currently available to detect piglet pain during castration and methods to mitigate castration-induced pain. In overview, the optimal outcome variables identified are nociceptive motor and vocal response scores during castration and quantitative sensory-threshold response testing and pain-associated behaviour scores following castration. Full article

(This article belongs to the Special Issue Refinements to Animal Models for Biomedical Research)

32 pages, 1585 KiB

Open AccessFeature PaperReview

The Impact of Common Recovery Blood Sampling Methods, in Mice (Mus Musculus), on Well-Being and Sample Quality: A Systematic Review

by Alexandra L Whittaker and Timothy H Barker

Animals 2020, 10(6), 989; https://doi.org/10.3390/ani10060989 - 5 Jun 2020

Cited by 15 | Viewed by 9069

Abstract

Blood sampling is often performed in laboratory mice. Sampling techniques have the potential to cause pain, distress and impact on lifetime cumulative experience. In spite of institutions commonly providing guidance to researchers on these methods, and the existence of published guidelines, no systematic evaluation of the evidence on this topic exists. A systematic search of Medline, Scopus, and Web of Science was performed, identifying 27 studies on the impact of recovery blood sample techniques on mouse welfare and sample quality. Studies were appraised for quality using the SYstematic Review Centre for Laboratory animal Experimentation (SYRCLE) risk of bias tool. In spite of an acceptable number of studies being located, few studies examined the same pairwise comparisons. Additionally, there was considerable heterogeneity in study design and outcomes, with many studies being at a high risk of bias. Consequently, results were synthesised using the Synthesis Without Meta-analysis (SWiM) reporting guidelines. Grading of Recommendations, Assessment, Development and Evaluation (GRADE) was utilised for assessment of certainty in the evidence. Due to the heterogeneity and GRADE findings, it was concluded that there was not enough high-quality evidence to make any recommendations on the optimal method of blood sampling. Future high-quality studies, with standardised outcome measures and large sample sizes, are required. Full article

(This article belongs to the Special Issue Refinements to Animal Models for Biomedical Research)

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Other

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22 pages, 924 KiB

Open AccessCommentary

An Anglocentric History of Anaesthetics and Analgesics in the Refinement of Animal Experiments

by R. Eddie Clutton

Animals 2020, 10(10), 1933; https://doi.org/10.3390/ani10101933 - 21 Oct 2020

Cited by 8 | Viewed by 5283

Abstract

Previous histories of animal experimentation, e.g., Franco (2013) have focused on ethics, the law and the personalities involved, but not on the involvement of anaesthetics or analgesics. Given that these were major subjects of (UK) Parliamentary debates on vivisection in the mid-19th century and viewed as “indisputable refinements in animal experimentation” (Russell and Burch 1959), it seemed that an analysis of their role was overdue. This commentary has, in interweaving the history of animal experimentation in the UK with the evolution of anaesthesia, attempted to: (1) clarify the evidence for Russell and Burch’s view; and (2) evaluate anaesthesia’s ongoing contribution to experimental refinement. The history that emerges reveals that the withholding or misuse of anaesthetics and, or analgesics from laboratory animals in the UK has had a profound effect on scientists and indirectly on the attitudes of the British public in general, becoming a major driver for the establishment of the anti-vivisection movement and subsequently, the Cruelty to Animals Act (1876)—the world’s first legislation for the regulation of animal experimentation. In 1902, the mismanaged anaesthetic of a dog in the Department of Physiology, University College London resulted in numerous events of public disorder initiated by medical students against the police and a political coalition of anti-vivisectionists, trade unionists, socialists, Marxists, liberals and suffragettes. The importance of anaesthesia in animal experiments was sustained over the following 150 years as small mammalian species gradually replaced dogs and cats as the principle subjects for vivisection. In discussing experimental refinement in their 1959 report, “The Principles of Humane Experimental Technique” Russell and Burch described anaesthetics as “… the greatest single advance in humane technique, (which) has at the same time been virtually indispensable for the advance of experimental biology”. Since then, the role of anaesthetics and in particular analgesics has become an unavoidable consideration whenever animal experiments are planned and conducted. This has been accompanied by a proliferation of training and educational programmes in laboratory animal anaesthesia. Full article

(This article belongs to the Special Issue Refinements to Animal Models for Biomedical Research)

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