Special Issue "The Welfare of Laboratory Animals"

A special issue of Animals (ISSN 2076-2615). This special issue belongs to the section "Animal Welfare".

Deadline for manuscript submissions: 31 October 2021.

Special Issue Editors

Prof. Dr. Lars Lewejohann
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Guest Editor
Freie Universität Berlin, Institute of Animal Welfare, Animal Behavior and Laboratory Animal Science, Berlin, Germany
Interests: animal welfare; refinement; animal behavior; laboratory animals; behavioral neuroscience; science of intelligence
Special Issues and Collections in MDPI journals
Prof. Dr. Christa Thöne-Reineke
E-Mail Website
Guest Editor
Freie Universität Berlin, Institute of Animal Welfare, Animal Behavior and Laboratory Animal Science, Berlin, Germany
Interests: animal welfare; animal behavior; laboratory animals and education
Special Issues and Collections in MDPI journals
Dr. Paulin Jirkof
E-Mail Website
Guest Editor
Department of Animals welfare, Zurich University, CH-8057 Zurich, Switzerland
Interests: animal welfare; 3Rs; animal behavior; pain assessment; severity assessment; analgesia; laboratory animals
Special Issues and Collections in MDPI journals
Prof. Dr. Helene Richter
E-Mail Website
Guest Editor
Department of Behavioural Biology, University of Münster, 48149 Münster, Germany
Interests: animal welfare; laboratory animals; 3R-concept with a main focus on the refinement, reproducibility and generalizability of animal experiments; individuality

Special Issue Information

Dear Colleagues,

The welfare of animals has become a growing societal concern. Using animals for experimental purposes is under particular scrutiny with regard to balancing the interests of scientific progress versus potential harm caused to the animals. Note that animal welfare concerns are not restricted to the experiment itself but span over the lifetime of the experimental animals from birth to death. Thus, better welfare of laboratory animals can be achieved by improving education in laboratory animal science, in the experimental design itself, but also by reliably assessing the welfare state of laboratory animals and improving the breeding, living, and/or handling conditions of the test animals before, during, and after the experiment. While mice and rats are currently the most frequently used species of laboratory animals, zebrafish as a model species are rising rapidly in numbers. Consequently, species-specific concepts have to be taken into account. The guiding principle in laboratory animal science is the 3R concept (replacement, reduction, refinement). Therefore, in addition to refinement strategies, replacement approaches and strategies for the reduction of animal numbers shall also be considered. This holds especially true with regard to surplus animals that are related to breeding and maintenance of the widely used model species or to unnecessary replication studies that are conducted because of a lack of knowledge transfer and transparency (e.g., publication bias).

This Special Issue is interested in both reviews and research papers on all aspects of laboratory animal welfare. We invite reports on the special requirements of model species as well as on more comprehensive general findings. We welcome strategies and concepts for the assessment of animal welfare and the severity of procedures involving animals, as well as case studies implementing practical improvement strategies. While there will naturally be a focus on refinement approaches, reduction and replacement strategies are also deemed suitable for this Special Issue. 

Prof. Dr. Lars Lewejohann
Prof. Dr. Christa Thöne-Reineke
Dr. Paulin Jirkof
Prof. Dr. Helene Richter
Guest Editors

Manuscript Submission Information

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 papers will be 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.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Animals is an international peer-reviewed open access monthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 1800 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • laboratory animals
  • mouse
  • rat
  • zebrafish
  • welfare assessment
  • 3R concept
  • refinement
  • housing conditions
  • communication
  • ethics
  • education

Published Papers (10 papers)

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Research

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Article
All the Pups We Cannot See: Cannibalism Masks Perinatal Death in Laboratory Mouse Breeding but Infanticide Is Rare
Animals 2021, 11(8), 2327; https://doi.org/10.3390/ani11082327 - 06 Aug 2021
Viewed by 562
Abstract
Perinatal mortality is a major issue in laboratory mouse breeding. We compared a counting method using daily checks (DAILY_CHECK) with a method combining daily checks with detailed video analyses to detect cannibalisms (VIDEO_TRACK) for estimating the number of C57BL/6 pups that were born, [...] Read more.
Perinatal mortality is a major issue in laboratory mouse breeding. We compared a counting method using daily checks (DAILY_CHECK) with a method combining daily checks with detailed video analyses to detect cannibalisms (VIDEO_TRACK) for estimating the number of C57BL/6 pups that were born, that died and that were weaned in 193 litters from trios with (TRIO-OVERLAP) or without (TRIO-NO_OVERLAP) the presence of another litter. Linear mixed models were used at litter level. To understand whether cannibalism was associated with active killing (infanticide), we analysed VIDEO_TRACK recordings of 109 litters from TRIO-OVERLAP, TRIO-NO_OVERLAP or SOLO (single dams). We used Kaplan-Meier method and logistic regression at pup level. For DAILY_CHECK, the mean litter size was 35% smaller than for VIDEO_TRACK (p < 0.0001) and the number of dead pups was twice lower (p < 0.0001). The risk of pup loss was higher for TRIO-OVERLAP than TRIO-NO_OVERLAP (p < 0.0001). A high number of pup losses occurred between birth and the first cage check. Analyses of VIDEO_TRACK data indicated that pups were clearly dead at the start of most of the cannibalism events and infanticide was rare. As most pups die and disappear before the first cage check, many breeding facilities are likely to be unaware of their real rates of mouse pup mortality. Full article
(This article belongs to the Special Issue The Welfare of Laboratory Animals)
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Communication
Large Farm Animals Used for Research Purposes: A Survey on Purchase, Housing and Hygiene Management
Animals 2021, 11(8), 2158; https://doi.org/10.3390/ani11082158 - 21 Jul 2021
Viewed by 778
Abstract
Background: Farm animals (FAs) are frequently used in biomedical research. Recommendations for the purchase, housing and health monitoring of these animals (sheep, goats, cattle and pigs) are still missing, and many institutes have developed their own strategies and protocols to face the challenges [...] Read more.
Background: Farm animals (FAs) are frequently used in biomedical research. Recommendations for the purchase, housing and health monitoring of these animals (sheep, goats, cattle and pigs) are still missing, and many institutes have developed their own strategies and protocols to face the challenges associated with the use of farm animals. This may influence the comparability of research results and increase data variances, thus increasing animal use that contradicts the obligation to apply the 3Rs principle of reduction, refinement and replacement required in Directive 2010/63 EU and the German animal protection law. Methods: A survey was conducted to define the current state of the art in research institutes working with pigs, and large and small ruminants. Results: The results of the survey clearly show that there are no uniform procedures regarding the purchase, housing and hygiene management of farm animals contrary to small laboratory animals. The facilities make purpose-bound decisions according to their own needs and individual work instructions and implement their own useful protocols to improve and maintain the health of the animals. Conclusion: This survey was the first step to filling the gaps and identifying the status quo and practical applied measures regarding the purchase and hygiene monitoring of FAs in order to improve animal welfare and scientific validity. Full article
(This article belongs to the Special Issue The Welfare of Laboratory Animals)
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Article
The Interactive Stress Assessment in Basic Animal Science Training
Animals 2021, 11(7), 2145; https://doi.org/10.3390/ani11072145 - 20 Jul 2021
Viewed by 804
Abstract
In order to assess the extent to which the legally prescribed training for the acquisition of animal experimentation expertise provides scientific personnel with the necessary competence and expertise to carry out a correct harm-benefit analysis in the context of animal experimentation applications, we [...] Read more.
In order to assess the extent to which the legally prescribed training for the acquisition of animal experimentation expertise provides scientific personnel with the necessary competence and expertise to carry out a correct harm-benefit analysis in the context of animal experimentation applications, we conducted an interactive stress assessment concerning the basic animal experimentation expertise course. First, before the practical part of the course and then, after the practical part, the participants assessed images and video material of healthy and stressed animals. The results were assessed comparatively and showed a significant increase in performance in all categories (p-value < 0.001). In addition, the results were comparatively assessed against those of scientists already experienced in animal experiments and experienced animal caretakers in research and clinics. In all groups, the vast majority of participants were able to recognise stress in laboratory animals. A significant proportion of the participants were also able to rate the level of stress correctly according to three degrees of severity: mild, moderate and severe. Nevertheless, a small number of participants were unable to distinguish between healthy and stressed animals and thus, the stress in the individual groups was assigned very differently from the different degrees of severity. The results of this study illustrate, on the one hand, the high significance that training must have in order to acquire the expertise, and, on the other hand, how strongly the assessment of stress is influenced by subjectivity. Full article
(This article belongs to the Special Issue The Welfare of Laboratory Animals)
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Article
Alternatives in Education—Rat and Mouse Simulators Evaluated from Course Trainers’ and Supervisors’ Perspective
Animals 2021, 11(7), 1848; https://doi.org/10.3390/ani11071848 - 22 Jun 2021
Viewed by 449
Abstract
Simulators allow the inexperienced to practice their skills prior to exercise on live animals. Therefore, they bear great potential in overcoming the dilemma between the present demand for high-quality practical training involving live animals whilst implementing the 3R principle according to the Directive [...] Read more.
Simulators allow the inexperienced to practice their skills prior to exercise on live animals. Therefore, they bear great potential in overcoming the dilemma between the present demand for high-quality practical training involving live animals whilst implementing the 3R principle according to the Directive 2010/63/EU. Currently, one mouse and six rat simulators are commercially available. As data on their impact are lacking, this project aimed at providing an overview of the awareness, implementation, and methodical and practical satisfaction provided by 35 course trainers and supervisors of laboratory animal training courses for mice and rats regarding the simulators available. Although simulators facilitate training of relevant techniques and relatively high awareness of them seemed to be present, their implementation is currently very low, possibly due to lack of meeting the respondents’ demands. Thus, this study revealed the overall approval of simulator training and general demand for user-optimized, realistic, and financially affordable simulators and, hence, indicates a strong impulse for new developments strengthening the 3Rs as a benefit to all animals used in research. Full article
(This article belongs to the Special Issue The Welfare of Laboratory Animals)
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Article
Additional Assessment of Fecal Corticosterone Metabolites Improves Visual Rating in the Evaluation of Stress Responses of Laboratory Rats
Animals 2021, 11(3), 710; https://doi.org/10.3390/ani11030710 - 05 Mar 2021
Viewed by 523
Abstract
Since animal experiments cannot be completely avoided, the pain, suffering, and distress of laboratory animals must be minimized. To this end, a major prerequisite is reliable assessment of pain and distress. Usually, evaluation of animal welfare is done by visual inspection and score [...] Read more.
Since animal experiments cannot be completely avoided, the pain, suffering, and distress of laboratory animals must be minimized. To this end, a major prerequisite is reliable assessment of pain and distress. Usually, evaluation of animal welfare is done by visual inspection and score sheets. However, relatively little is known about whether standardized, but subjective, score sheets are able to reliably reflect the status of the animals. The current study aimed to compare visual assessment scores and changes in body weight with concentrations of fecal corticosterone metabolites (FCMs) in a neuroscientific experimental setup. Additionally, effects of refinement procedures were investigated. Eight male adult Sprague-Dawley rats underwent several experimental interventions, including electroencephalograph electrode implantation and subsequent recording, positron emission tomography (PET), and sleep deprivation (SD) by motorized activity wheels. Additional 16 rats were either used as controls without any treatment or to evaluate refinement strategies. Stress responses were determined on a daily basis by means of measuring FCMs, body weight, and evaluation of the animals’ welfare by standardized score sheets. Surgery provoked a significant elevation of FCM levels for up to five days. Increases in FCMs due to PET procedures or SD in activity wheels were also highly significant, while visual assessment scores did not indicate elevated stress levels and body weights remained constant. Visual assessment scores correlate with neither changes in body weight nor increases in FCM levels. Habituation procedures to activity wheels used for SD had no impact on corticosterone release. Our results revealed that actual score sheets for visual assessment of animal welfare did not mirror physiological stress responses assessed by FCM measurements. Moreover, small changes in body weight did not correlate with FCM concentration either. In conclusion, as visual assessment is a method allowing immediate interventions on suffering animals to alleviate burden, timely stress assessment in experimental rodents via score sheets should be ideally complemented by validated objective measures (e.g., fecal FCM measured by well-established assays for reliable detection of FCMs). This will complete a comprehensive appraisal of the animals’ welfare status in a retrospective manner and refine stressor procedures in the long run. Full article
(This article belongs to the Special Issue The Welfare of Laboratory Animals)
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Article
Isoflurane and Carbon Dioxide Elicit Similar Behavioral Responses in Rats
Animals 2020, 10(8), 1431; https://doi.org/10.3390/ani10081431 - 16 Aug 2020
Viewed by 1271
Abstract
Euthanasia in rodents is an ongoing topic of debate due to concerns regarding the aversive nature of gases with anesthetic properties such as carbon dioxide (CO2) and isoflurane. The aim of this study was to expand upon previously published work evaluating [...] Read more.
Euthanasia in rodents is an ongoing topic of debate due to concerns regarding the aversive nature of gases with anesthetic properties such as carbon dioxide (CO2) and isoflurane. The aim of this study was to expand upon previously published work evaluating the aversiveness of CO2 by introducing an isoflurane treatment group in parallel. Aversion was tested using a forced exposure setup and an aversion-avoidance setup. In the first part of the study, 12 naïve female Sprague–Dawley rats were exposed during four consecutive days, once to each of four treatments: isoflurane, fox urine, oxygen, and CO2. In the second part of the study, 24 naïve female Sprague–Dawley rats and 12 rats from the first experiment were exposed to CO2, isoflurane, or both gases. In the forced exposure study, there were no significant differences between CO2 and isoflurane treatments except in line crosses. Overall, rats were more active in the isoflurane and CO2 treatments compared to the control groups, suggesting that isoflurane and CO2 are similarly aversive. In the aversion-avoidance study, rats previously exposed to isoflurane left the dark chamber significantly earlier compared to naïve rats during exposure to isoflurane. We also show that learned aversion to isoflurane is sustained for at least 15 days after initial exposure. Given this result, we suggest that CO2 is superior to isoflurane when euthanizing rodents with prior exposure to isoflurane. Overall, these results confirm previous studies which suggest that care should be taken when considering the serial use of isoflurane as an anesthetic. Full article
(This article belongs to the Special Issue The Welfare of Laboratory Animals)
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Review

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Review
Developing Recommendations for Cumulative Endpoints and Lifetime Use for Research Animals
Animals 2021, 11(7), 2031; https://doi.org/10.3390/ani11072031 - 07 Jul 2021
Viewed by 1037
Abstract
Research animals are important for scientific advancement, and therefore, their long-term welfare needs to be monitored to not only minimize suffering, but to provide positive affective states and experiences. Currently, there is limited guidance in countries around the world on cumulative and experimental [...] Read more.
Research animals are important for scientific advancement, and therefore, their long-term welfare needs to be monitored to not only minimize suffering, but to provide positive affective states and experiences. Currently, there is limited guidance in countries around the world on cumulative and experimental endpoints. This paper aims to explore current opinions and institutional strategies regarding cumulative use and endpoints through a scoping survey and review of current regulations and welfare assessment tools, and ultimately to provide recommendations for assessment of cumulative and lifetime use of research animals. The survey found that only 36% of respondents indicated that their institution had cumulative use endpoint policies in place, but these policies may be informal and/or vary by species. Most respondents supported more specific guidelines but expressed concerns about formal policies that may limit their ability to make case-by-case decisions. The wide diversity in how research animals are used makes it difficult for specific policies to be implemented. Endpoint decisions should be made in an objective manner using standardized welfare assessment tools. Future research should focus on robust, efficient welfare assessment tools that can be used to support planning and recommendations for cumulative endpoints and lifetime use of research and teaching animals. Full article
(This article belongs to the Special Issue The Welfare of Laboratory Animals)
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Review
Enrichment for Laboratory Zebrafish—A Review of the Evidence and the Challenges
Animals 2021, 11(3), 698; https://doi.org/10.3390/ani11030698 - 05 Mar 2021
Cited by 2 | Viewed by 1986
Abstract
Good practice for the housing and care of laboratory zebrafish Danio rerio is an increasingly discussed topic, with focus on appropriate water quality parameters, stocking densities, feeding regimes, anaesthesia and analgesia practices, methods of humane killing, and more. One area of current attention [...] Read more.
Good practice for the housing and care of laboratory zebrafish Danio rerio is an increasingly discussed topic, with focus on appropriate water quality parameters, stocking densities, feeding regimes, anaesthesia and analgesia practices, methods of humane killing, and more. One area of current attention is around the provision of environmental enrichment. Enrichment is accepted as an essential requirement for meeting the behavioural needs and improving the welfare of many laboratory animal species, but in general, provision for zebrafish is minimal. Some of those involved in the care and use of zebrafish suggest there is a ‘lack of evidence’ that enrichment has welfare benefits for this species, or cite a belief that zebrafish do not ‘need’ enrichment. Concerns are also sometimes raised around the practical challenges of providing enrichments, or that they may impact on the science being undertaken. However, there is a growing body of evidence suggesting that various forms of enrichment are preferred by zebrafish over a barren tank, and that enriched conditions can improve welfare by reducing stress and anxiety. This review explores the effects that enrichment can have on zebrafish behaviour, physiology and welfare, and considers the challenges to facilities of providing more enrichment for the zebrafish they house. Full article
(This article belongs to the Special Issue The Welfare of Laboratory Animals)
Review
Methods Used and Application of the Mouse Grimace Scale in Biomedical Research 10 Years on: A Scoping Review
Animals 2021, 11(3), 673; https://doi.org/10.3390/ani11030673 - 03 Mar 2021
Cited by 2 | Viewed by 769
Abstract
The Mouse Grimace Scale (MGS) was developed 10 years ago as a method for assessing pain through the characterisation of changes in five facial features or action units. The strength of the technique is that it is proposed to be a measure of [...] Read more.
The Mouse Grimace Scale (MGS) was developed 10 years ago as a method for assessing pain through the characterisation of changes in five facial features or action units. The strength of the technique is that it is proposed to be a measure of spontaneous or non-evoked pain. The time is opportune to map all of the research into the MGS, with a particular focus on the methods used and the technique’s utility across a range of mouse models. A comprehensive scoping review of the academic literature was performed. A total of 48 articles met our inclusion criteria and were included in this review. The MGS has been employed mainly in the evaluation of acute pain, particularly in the pain and neuroscience research fields. There has, however, been use of the technique in a wide range of fields, and based on limited study it does appear to have utility for pain assessment across a spectrum of animal models. Use of the method allows the detection of pain of a longer duration, up to a month post initial insult. There has been less use of the technique using real-time methods and this is an area in need of further research. Full article
(This article belongs to the Special Issue The Welfare of Laboratory Animals)
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Other

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Commentary
Breeding and Maintenance of Immunodeficient Mouse Lines under SPF Conditions—A Call for Individualized Severity Analyses and Approval Procedures
Animals 2021, 11(6), 1789; https://doi.org/10.3390/ani11061789 - 15 Jun 2021
Viewed by 1102
Abstract
In the EU, the breeding of genetically modified laboratory animals is, by definition, an animal experiment if the offspring may experience pain, suffering, or harm. In order to determine the actual burden of genetically modified mice, established methods are available. However, the breeding [...] Read more.
In the EU, the breeding of genetically modified laboratory animals is, by definition, an animal experiment if the offspring may experience pain, suffering, or harm. In order to determine the actual burden of genetically modified mice, established methods are available. However, the breeding of immunodeficient mice is considered an experiment requiring a permit, even if no pain, suffering or harm is observed under scientifically required defined hygienic housing conditions, as determined by established methods of severity assessment. This seems contradictory and leads to uncertainty among scientists. With this commentary, we would like to shed light on this topic from different perspectives and propose a solution in terms of individualized severity assessment and approval procedures. Full article
(This article belongs to the Special Issue The Welfare of Laboratory Animals)
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