7 Nov 2019, 4:47 am
Laboratory Animals
Laboratory Animals, Ahead of Print.
Evidence-based severity assessment is essential as a basis for ethical evaluation in animal experimentation to ensure animal welfare, legal compliance and scientific quality. To fulfil these tasks scientists, animal care and veterinary personnel need assessment tools that provide species-relevant measurements of the animals' physical and affective state. In a three-centre study inter-laboratory robustness of body weight monitoring, mouse grimace scale (MGS) and burrowing test were evaluated. The parameters were assessed in naïve and tramadol treated female C57BL/6J mice. During tramadol treatment a body weight loss followed by an increase, when treatment was terminated, was observed in all laboratories. Tramadol treatment did not affect the MGS or burrowing performance. Results were qualitatively comparable between the laboratories, but quantitatively significantly different (inter-laboratory analysis). Burrowing behaviour seems to be highly sensitive to inter-laboratory differences in testing protocol. All locations obtained comparable information regarding the qualitative effect of tramadol treatment in C57BL/6J mice, however, datasets differed as a result of differences in test and housing conditions.In conclusion, our study confirms that results of behavioural testing can be affected by many factors and may differ between laboratories. Nevertheless, the evaluated parameters appeared relatively robust even when conditions were not harmonized extensively and present useful tools for severity assessment. However, analgesia-related side effects on parameters have to be considered carefully.
31 Oct 2019, 6:04 pm
Laboratory Animals
Laboratory Animals, Ahead of Print.
Voluntary wheel running (VWR) behaviour is a sensitive indicator of disturbed wellbeing and used for the assessment of individual experimental severity levels in laboratory mice. However, monitoring individual VWR performance usually requires single housing, which itself might have a negative effect on wellbeing. In consideration of the 3Rs principle, VWR behaviour was evaluated under group-housing conditions. To test the applicability for severity assessment, this readout was evaluated in a dextran sodium sulphate (DSS) induced colitis model. For continuous monitoring, an automated system with integrated radio-frequency identification technology was used, enabling detection of individual VWR. After a 14-day adaptation period mice demonstrated a stable running performance. Analysis during DSS treatment in combination with repeated facial vein phlebotomy and faecal sampling procedure resulted in significantly reduced VWR behaviour during the course of colitis and increased VWR during disease recovery. Mice submitted to phlebotomy and faecal sampling but no DSS treatment showed less reduced VWR but a longer-lasting recovery. Application of a cluster model discriminating individual severity levels based on VWR and body weight data revealed the highest severity level in most of the DSS-treated mice on day 7, but a considerable number of control mice also showed elevated severity levels due to sampling procedures alone. In summary, VWR sensitively indicated the course of DSS colitis severity and the impact of sample collection. Therefore, monitoring of VWR is a suitable method for the detection of disturbed wellbeing due to DSS colitis and sampling procedure in group-housed female laboratory mice.
30 Oct 2019, 10:09 pm
Laboratory Animals
Laboratory Animals, Ahead of Print.
In many animal experiments scientists and local authorities define a body-weight reduction of 20% or more as severe suffering and thereby as a potential parameter for humane endpoint decisions. In this study, we evaluated distinct animal experiments in multiple research facilities, and assessed whether 20% body-weight reduction is a valid humane endpoint criterion in rodents. In most experiments (restraint stress, distinct models for epilepsy, pancreatic resection, liver resection, caloric restrictive feeding and a mouse model for Dravet syndrome) the animals lost less than 20% of their original body weight. In a glioma model, a fast deterioration in body weight of less than 20% was observed as a reliable predictor for clinical deterioration. In contrast, after induction of chronic diabetes or acute colitis some animals lost more than 20% of their body weight without exhibiting major signs of distress. In these two animal models an exclusive application of the 20% weight loss criterion for euthanasia might therefore result in an unnecessary loss of animals. However, we also confirmed that this criterion can be a valid parameter for defining the humane endpoint in other animal models, especially when it is combined with additional criteria for evaluating distress. In conclusion, our findings strongly suggest that experiment and model specific considerations are necessary for the rational integration of the parameter ‘weight loss’ in severity assessment schemes and humane endpoint criteria. A flexible implementation tailored to the experiment or intervention by scientists and authorities is therefore highly recommended.
28 Oct 2019, 6:01 pm
Laboratory Animals
Laboratory Animals, Ahead of Print.
The Mouse Grimace Scale (MGS) is an established method for estimating pain in mice during animal studies. Recently, an improved and standardized MGS set-up and an algorithm for automated and blinded output of images for MGS evaluation were introduced. The present study evaluated the application of this standardized set-up and the robustness of the associated algorithm at four facilities in different locations and as part of varied experimental projects. Experiments using the MGS performed at four facilities (F1–F4) were included in the study; 200 pictures per facility (100 pictures each rated as positive and negative by the algorithm) were evaluated by three raters for image quality and reliability of the algorithm. In three of the four facilities, sufficient image quality and consistency were demonstrated. Intraclass correlation coefficient, calculated to demonstrate the correlation among raters at the three facilities (F1–F3), showed excellent correlation. The specificity and sensitivity of the results obtained by different raters and the algorithm were analysed using Fisher's exact test (p < 0.05). The analysis indicated a sensitivity of 77% and a specificity of 64%. The results of our study showed that the algorithm demonstrated robust performance at facilities in different locations in accordance with the strict application of our MGS setup.
28 Oct 2019, 6:01 pm
Laboratory Animals
Laboratory Animals, Ahead of Print.
According to European Union directive 2010/63/EU a severity classification of experimental procedures performed on laboratory animals is mandatory. This includes a prospective evaluation of all interventions performed within the experiment, as well as an assessment of the actual burden of each animal during the experiment. In this regard, the evaluation and scoring of defined criteria regarding the health state of animals could help to early identify deteriorations in animal health and facilitate the application of humane endpoints. This article discusses the applicability of an adapted score sheet in BALB/cAnNRj mice receiving either cisplatin, doxorubicin or busulfan, three chemotherapeutic agents with different toxicological profiles and longitudinal non-invasive molecular imaging. The health state was investigated by score sheets documenting general state, body weight, spontaneous behaviour and treatment specific parameters (e.g. anaemia, neurotoxicity, persistent diarrhoea). Although blood and serum analyses clearly indicated various organ damage, most scoring parameters except for body weight did not report on the deceasing animal health state. Thus, there is need for more sensitive observational parameters to judge the animal's health state and welfare.
25 Oct 2019, 6:31 pm
Laboratory Animals
Laboratory Animals, Ahead of Print.
Severity assessment for experiments conducted with laboratory animals is still based mainly on subjective evaluations; evidence-based methods are scarce. Objective measures, amongst which determination of the concentrations of stress hormones, can be used to aid severity assessment. Short-term increases in glucocorticoid concentrations generally reflect healthy responses to stressors, but prolonged increases may indicate impaired welfare. As mice are the most commonly used laboratory animal species, we performed a systematic mapping review of corticosterone measurements in Mus musculus, to provide a full overview of specimen types (e.g. blood, urine, hair, saliva, and milk) and analysis techniques. In this publication, we share our protocol and search strategy, and our rationale for performing this systematic analysis to advance severity assessment. So far, we have screened 13,520 references, and included 5337 on primary studies with measurements of endogenous corticosterone in M. musculus. Data extraction is currently in progress. When finished, this mapping review will be a valuable resource for scientists interested in corticosterone measurements to aid severity assessment. We plan to present the data in a publication and a searchable database, which will allow for even easier retrieval of the relevant literature. These resources will aid implementation of objective measures into severity assessment.
24 Oct 2019, 10:33 pm
Laboratory Animals
Laboratory Animals, Ahead of Print.
The Mouse Grimace Scale (MGS) has been widely used for the noninvasive examination of distress/pain in mice. The aim of this study was to further improve its performance to generate repeatable, faster, blinded and reliable results for developing automated and standardized pictures for MGS scoring and simultaneous evaluation of up to four animals.Videos of seven C57BL/6N mice were generated in an experiment to assess pain and stress induced by repeated intraperitoneal injection of carbon tetrachloride (CCl4). MGS scores were taken 1 h before and after the injection. Videotaping was performed for 10 min in special observation boxes. For manual selection, pictures of each mouse were randomly chosen for quality analysis and scored according six quality selection criteria (0 = no, 1 = moderate, 2 = full accordance); the maximum possible score was 12.Overall, 609 pictures from six videos were evaluated for MGS scoring quality; evaluation was performed by using the picture selection tool or by manual scoring. With manual scoring, 288 pictures (48.3% of all randomly generated pictures) were deemed scorable using MGS (mean score = 22.15 ± SD 6.3). To evaluate the algorithm, ratings from different rater groups (beginner, medium-level trained, professional) were compared with the automated image generated. These differences were not significant (p = 0.1091).This study demonstrates an improved set-up and a picture selection tool that can generate repeatable, not-observer biased and standardized pictures for MGS scoring.
24 Oct 2019, 10:33 pm
Laboratory Animals
Laboratory Animals, Ahead of Print.
The pig is commonly used in renal transplantation studies since the porcine kidney resembles the human kidney. To meet the requirements of intense caretaking and examination without stress, a 2-week socialisation and training programme was developed. Conventional cross-breed pigs (n = 36) with high health status were trained for 15 min/day in a four-step training programme before kidney transplantation. The systematic training resulted in calm animals, which allowed for ultrasound examination, blood sampling and urine sampling without restraint. When a 2-methacryloyloxyethyl phosphorylcholine polymer-coated jugular catheter introduced via the auricular vein was used for post-operative blood sampling, clotting was avoided. To assess renal function, urinary output was observed and creatinine and cystatin C were measured; the latter was not found to be useful in recently transplanted pigs. The results presented contribute to the 3Rs (refine, reduce, replace).
20 Oct 2019, 10:07 pm
Laboratory Animals
Laboratory Animals, Ahead of Print.
Different online courses and training programs in Laboratory Animal Science (LAS) have emerged across Europe in recent years. E-learning appears to be a promising solution to achieve flexibility in training while meeting the quality criteria of demanding programs in short training periods. However, little is known about how students perceive e-learning in this context, and there is also a lack of specific and valid instruments to measure this perception. Within an exploratory study framework, the e-learning perception of 229 participants in 15 courses in Portugal using two different online training formats, flipped classroom and full online theoretical training, was assessed. For this purpose, the Questionnaire of E-learning Acceptance (QELA), a 32-item accordance Likert-type scale comprising five subscales was developed to explore the following: how participant perceive e-learning, satisfaction with organization and contents, perception of e-learning relevance for the time management, and its influence for practical training. In general, e-learning was well accepted and perceived to work well and be useful by the majority of courses participants, independently of the course level and e-learning format approach. These results indeed suggest that integration of e-learning is useful in LAS training. We also propose the QELA as a starting point for development and implementation of specific instruments to assess e-learning acceptance in LAS across a wider range of geographical and training contexts.
9 Oct 2019, 10:10 pm
Laboratory Animals
Laboratory Animals, Ahead of Print.
The objective was to determine the rate at which Chinese journals include Animal Research: Reporting of In Vivo Experiments (ARRIVE) Guidelines/Gold Standard Publication Checklist (GSPC) in their instructions for authors, and the awareness and recognition of editors. The survey was performed on Chinese journals. The most recent versions each journal's instructions for authors were downloaded, and the information related to the ARRIVE/GSPC was collected. A self-developed questionnaire was used to conduct the survey among the editors. Questionnaires were sent to 238 qualified journals and 198 of them returned them, achieving an 83.2% response rate. The results showed that none of the journals included the ARRIVE/GSPC in their instructions for authors, and the awareness rate was only 13.1% (26/198). The participants who were unaware of the ARRIVE/GSPC were less likely than those who were aware of them to believe it was necessary to include the ARRIVE/GSPC in the instructions for authors (23.3% vs. 61.5%), and less likely to request authors in their manuscript preparation (28.5% vs. 88.5%), editors in the editing and processing (28.5% vs. 84.6%) and reviewers in peer review stage (28.5% vs. 92.3%) to follow the ARRIVE/GSPC. Currently no Chinese journals include the ARRIVE/GSPC in their instructions for authors. The recognition rate of the ARRIVE/GSPC was low among the editors. So, we suggest that Chinese journals should promote inclusion of the ARRIVE/GSPC in journals' instructions for authors. It is also important to educate researchers and editors alike to increase their understanding of the ARRIVE/GSPC, so that the quality of reporting of animal study can be improved.