Case-based teaching of experimental design - contributions for meaningful learning.

This article argues the need for education and training of researchers carrying out animal studies on the fundamentals of experimental design (ED), as a key means of improving the reliability and reproducibility of preclinical results. The current landscape in ED education in Europe is presented, and we make the case for dedicated tutor-guided teaching of ED. With less than a day dedicated to it in many courses effective techniques for communicating key issues are needed. We have developed two approaches that transfer to experimental design teaching the case-study, problem-solving techniques known to be effective in other fields. They use realistic research scenarios to provoke discussion and engage learning. In one the scenario is for group discussion or informal or formal assessment with subsequent tutor-led discussion of key points. For this each scenario needs a clear statement of the purpose of the research study, simplified text outlining the comparisons and procedures, and a statement of the outcome measure. In the other approach, the scenario is used with freely-available software with a good graphical output to explore the sizing of experiments and the use of both sexes. Trainee feedback and informal assessment show that these approaches can make for interesting and memorable sessions and offer a useful contribution to improvement in experimental design teaching so that it produces meaningful learning that can translate into better practice.

Methodological standards, quality of reporting and regulatory compliance in animal research on amyotrophic lateral sclerosis: a systematic review.

OBJECTIVES: The amyotrophic lateral sclerosis (ALS) research community was one of the first to adopt methodology guidelines to improve preclinical research reproducibility. We here present the results of a systematic review to investigate how the standards in this field changed over the 10-year period during which the guidelines were first published (2007) and updated (2010). METHODS: We searched for papers reporting ALS research on SOD1 (superoxide dismutase 1) mice published between 2005 and 2015 on the ISI Web of Science database, resulting in a sample of 569 papers to review, after triage. Two scores-one for methodological quality, one for regulatory compliance-were built from weighted sums of separate sets of items, and subjected to multivariable regression analysis, to assess how these related to publication year, type of study, country of origin and journal. RESULTS: Reporting standards improved over time. Of papers published after the first ALS guidelines were made public, fewer than 9% referred specifically to these. Of key research parameters, only three (genetic background, number of transgenes and group size) were reported in >50% of the papers. Information on housing conditions, randomisation and blinding was absent in over two-thirds of the papers. Group size was among the best reported parameters, but the majority reported using fewer than the recommended sample size and only two studies clearly justified group size. CONCLUSIONS: General methodological standards improved gradually over a period of 8-10 years, but remained generally comparable with related fields with no specific guidelines, except with regard to severity. Only 11% of ALS studies were classified in the highest severity level (animals allowed to reach death or moribund stages), substantially below the proportion in studies of comparable neurodegenerative diseases such as Huntington's. The existence of field-specific guidelines, although a welcome indication of concern, seems insufficient to ensure adherence to high methodological standards. Other mechanisms may be required to improve methodological and welfare standards.

Eye, body or tail? Thermography as a measure of stress in mice.

Infrared thermography has been suggested as a non-invasive, objective tool to evaluate animal welfare. In this study, we investigated: 1) how body temperature, measured through thermal imaging, is affected by different mild stressors frequently experienced by laboratory mice; 2) which methodology to use for assessing temperature variations with infrared thermography; 3) whether the chosen stressors cause anxiety in mice. Eighty C57BL/6 male mice were included in the study. The mice were allocated to either a control group or one of three groups being subjected to a mild stressor once daily for 4 days: 1) anaesthesia with isoflurane for 10 min; 2) handling by scruffing; 3) intraperitoneal injection of 0.2 ml 0.9% saline. On all four intervention days, thermal images were obtained in all groups and all animals were assessed for fur status and body weight. On day five, all animals were tested in the elevated-plus-maze for 5 min. From the thermal images, the maximum eye temperature, the maximum tail base temperature and the average body temperature were obtained. Ten minutes of anaesthesia with isoflurane led to a decrease in maximum eye temperature, average body temperature and maximum tail base temperature. The animals recovered from this drop in temperature within 10 min. No drop in temperature was seen after scruffing or intraperitoneal injection of saline. Based on the number of missing values, intra-rater and inter-rater agreement, the average body temperature was found most ideal for measuring body temperature variations in mice. Finally, the elevated plus maze did not reveal any differences in anxiety between the groups and the body weight did not decrease at any time point during the study.

"How sick must your mouse be? " - An analysis of the use of animal models in Huntington's disease research.

Refinement measures to improve animal welfare can ease the ethical dilemma between human benefit and animal harm in research with animal models of neurodegenerative diseases. To evaluate the potential for refinement, as well as its implementation in research, we analysed papers on murine models of Huntington's disease (HD) published between 1999 and 2009 (n = 233). Each study was classified according to a four-level severity scale, in terms of the disease stage that animals were allowed to reach, the execution of invasive procedures, and the implementation of refinement. Reports of ethical approval, and regulatory compliance in general, followed a clear rising trend over the years reviewed (p <0.001). However, the proportion of high-severity studies did not change over that period. Also, of the studies for which approval was reported (n = 120), 36% were attributed the highest severity level. The observed discrepancy between the rising motivation to affirm regulatory compliance, and the unaltered proportion of studies allowing animals to reach severely distressful stages, raises both ethical and methodological issues, which we discuss in this paper.