M. Ritskes-Hoitinga

The Gold Standard Publication Checklist (GSPC) for improved design, reporting and scientific quality of animal studies GSPC versus ARRIVE guidelines

Hundreds of experiments in which animals are used to answer biomedical research questions are performed and published every month. Although well-designed and performed animal experiments are a necessary condition for successful translational research, many papers involving animal experimentation are still incomplete in their reporting.1–4 Clearly, there is an urgent need to improve the reporting of animal experiments in order to increase the scientific quality of animal studies, animal welfare and ultimately patient safety.5–7 Currently, the paradoxical situation exists that the high standards set for clinical trials are not applied in animal experiments, even though these animal studies are performed with the aim to improve human health care. Against this background, the ARRIVE guidelines were published in PLoS Biology in July 2010.8 We strongly support this initiative and believe it will make an important contribution to improving the reporting of animal studies. n nIn our view, guidelines are not only necessary for increasing the quality of reporting of completed animal studies, but are also essential for optimal design and execution of new animal experiments, and thus improved scientific quality. With these goals in mind, we developed the Gold Standard Publication Checklist (GSPC): Hooijmans et al. A gold standard publication checklist to improve the quality of animal studies, to fully integrate the three Rs, and to make systematic reviews more feasible. Altern Lab Anim 2010;38(2):167–82. This checklist was presented and discussed at the World Congress on alternatives and animal use in the life sciences in Rome in 2009 and published in ATLA in May 2010, a few months before the ARRIVE guidelines appeared. n nGiven their partly similar aims, the GSPC has some overlap with the ARRIVE guidelines. However, the GSPC describes certain items in more detail, for instance the housing conditions (humidity, ventilation, lighting, noise, caging), nutrition (type of diet, diet content, method of feeding) and water. These detailed descriptions in the checklist help scientists to include all the specific items necessary for planning, designing and performing animal experiments in the most optimal way, and to improve repeatability of and control variation within experiments, through which the quality of research improves and the number of animals needed in an experiment diminishes. In addition, the GSPC paper highlights the importance of reporting husbandry conditions and basic principles of the design of animal experiments by providing an overview of the literature on how and when interference with experimental results may occur. Last but not least, the GSPC is presented as a checklist, and therefore well-ordered and easy to use when designing and executing animal experiments. n nThe use of guidelines for designing, executing and reporting of animal experiments (like the ARRIVE guidelines or the GSPC) will also make systematic reviews (SRs) and meta-analyses of publications on animal studies more feasible.9,10 SRs can be defined as a literature review focused on a single question that tries to identify, appraise, select and synthesize all available high-quality research evidence relevant to that question. These SRs lead to better interpretation of the already existing scientific results from animal experiments, through which a better translation to the clinic and more guarantees for patient safety become reality. Furthermore, unnecessary duplication of animal experiments, and thereby unnecessary animal use and time loss, will be prevented. SRs are already standard practice in clinical studies and it is about time that they will become standard practice in the field of animal studies as well.3,11 n nTo conclude, in order to make SRs feasible and to improve not only the reporting but also the planning, design and execution of animal studies, we strongly recommend all scientists involved in animal experimentation and editors of journals publishing animal studies to make use of the GSPC and/or the ARRIVE guidelines.

An initiative to facilitate well-informed decision-making in laboratory animal research: report of the First International Symposium on Systematic Reviews in Laboratory Animal Science:

Most of us are overloaded with information. Even when we have time to read some of the information that arrives on our computers daily, it is difficult to identify which information will be most useful and to recall the most up-to-date findings when we need them. This is where systematic reviews come in. A systematic review focuses on a specific research question and tries to identify, appraise, select and synthesize all high-quality research evidence relevant to the question. These reviews are standard practice in clinical studies, but are not yet widely conducted in the field of laboratory animal science. Given that many studies using laboratory animals are aimed at improving human health (and health care), it seems logical that research using animals is reviewed in a similar way and adheres to similar high-quality standards. Apparently, this is not the case yet. In order to highlight the importance of systematic reviews for the field of laboratory animal science, we launched the First International Symposium on Systematic Reviews in Laboratory Animal Science in Nijmegen (The Netherlands).

The influence of food restriction versus ad libitum feeding of chow and purified diets on variation in body weight, growth and physiology of female Wistar rats:

Ad libitum (AL) supply of standard chow is the feeding method most often used for rodents in animal experiments. However, AL feeding is known to result in a shorter lifespan and decreased health as compared with restricted feeding. Restricted feeding and thus limiting calorie intake prevents many health problems, increases lifespan and can also increase group uniformity. All this leads to a reduced number of animals needed. So-called standard chows are known to be prone to variation in composition. Synthetic foods have a more standard composition, contributing to group uniformity which, like diet reduction, may decrease the number of animals necessary to obtain statistical significance. In this study, we compared the effects of AL versus restricted feeding (25% reduction in food intake) on standard chow versus synthetic food of three different suppliers on body weight (BW), growth, several blood parameters and organ weights in growing female Wistar rats over a period of 61 days. Diet restriction led to a decreased growth and significantly reduced variation in BW and growth as compared with AL feeding. AL feeding on synthetic diets caused a significantly higher BW gain than on chow diets. Due to experimental design, this same effect occurred on food restriction. Blood parameters and organ weights were affected neither by diet type nor by amount. Incidentally, variations were significantly reduced on food restriction versus AL, and on synthetic diets versus chow diets. This study demonstrates that food restriction versus AL feeding leads to a significantly reduced variation in BW and growth, thereby indicating the potential for reduction when applying this feeding schedule.

Renal epithelial proliferation and its clinical expression in Brown Norway (BN) rats

Renal epithelial proliferation has previously been found to be a common condition in a colony of Lewis × Brown Norway (BN) F2 hybrid rats. The aim of this study was to investigate the prevalence and clinical consequences of this condition in pure inbred BN and Lewis rats. Renal epithelial proliferation was found in 29 of 49 BN rats (59%) examined and in four of 50 Lewis rats (8%) examined. Serum creatinine and serum corticosterone was not influenced by the condition. Haematuria was more common in BN rats with (74%) than without renal papillary proliferation (35%, P < 0.05), but it may not be used to diagnose renal epithelial proliferation, as we found rats having renal epithelial proliferation without showing haematuria and rats showing haematuria without having renal epithelial proliferation. Haematuria was also common in Lewis rats (16-56% dependent of age and gender), in which renal epithelial proliferation were found in only 8%. Fluctuating asymmetry, which was used as a measure of developmental instability, was found to be increased in rats with renal epithelial proliferation (P < 0.05). Haematuria was also found to be related to the degree of fluctuating asymmetry (P < 0.01). Although the prevalence of renal epithelial proliferation is clearly higher in BN rats than in Lewis rats (P < 0.01), and although in previous reports the condition was found in F2 BN × Lewis hybrids and not in F1 BN × Lewis hybrids it cannot clearly be defined as having been caused by a single Mendelian gene, as we found it in both inbred strains. Futhermore, we found that morphologically the proliferations could be placed on the papillary as well as the medullary wall of the renal pelvis, while previously it has only been described on the papillary wall.

Ibogaine and addiction in the animal model, a systematic review and meta-analysis

Ibogaine is a naturally occurring substance which has been increasingly used in the lay-scene to reduce craving and relapse in patients with substance use disorders (SUDs). Although human clinical trials on the safety and efficacy of ibogaine are lacking, animal studies do support the efficacy of ibogaine. In this systematic review and meta-analysis (MA), we summarise these animal findings, addressing three questions: (1) does ibogaine reduce addictive behaviour in animal models of SUDs?; (2) what are the toxic effects of ibogaine on motor functioning, cerebellum and heart rhythm?; (3) what are neuropharmacological working mechanisms of ibogaine treatment in animal models of SUDs? MA of 27 studies showed that ibogaine reduced drug self-administration, particularly during the first 24u2009h after administration. Ibogaine had no effect on drug-induced conditioned place preference. Ibogaine administration resulted in motor impairment in the first 24u2009h after supplementation, and cerebral cell loss even weeks after administration. Data on ibogaines effect on cardiac rhythm, as well as on its neuropharmacological working mechanisms are limited. Our results warrant further studies into the clinical efficacy of ibogaine in SUD patients in reducing craving and substance use, but close monitoring of the patients is recommended because of the possible toxic effects. In addition, more work is needed to unravel the neuropharmacological working mechanisms of ibogaine and to investigate its effects on heart rhythm.

Outcomes of a Dutch workshop on improvements for the 3Rs in daily practice

This article describes the outcome of a workshop that was held to generate new ideas to improve the use of the 3R principles in science. The participants of the workshop represented Dutch researchers, Animal Welfare Officers, and members of Animal Ethics Committees from various affiliations, including academia, industry, contract research organizations, and knowledge centers in the Netherlands. The workshop resulted in six diverse consensus statements, which are presented and discussed in this article. The results show that there is no single, straightforward solution to improving the implementation of 3R methods in animal-based research and that further improvement in 3Rs practice requires a more multi- and interdisciplinary awareness and approach.

Improving the conduct, reporting, and appraisal of animal research

All stakeholders must act decisively to fix endemic problems

An interview study of phenotypic characterization of genetically-modified mice

An interview study was carried out with the aim of clarifying the reasons for the limited use of phenotypic characterization of genetically-modified mice (GMM) and identifying issues hindering its implementation. A total of 15 users of GMM participated in semi-structured face-to-face interviews, which were audio-taped and transcribed. The results were extracted using content analysis by theme. The investigation confirmed that few animals were systematically phenotyped and an observational approach was found to be widespread. The primary interest of the interviewees was phenotyping for impaired animal welfare. The concept of phenotyping was widely understood and perceived as a scientific advantage. The comprehensiveness of the protocols and the resources required for phenotyping were seen as problematic. All participants addressed this issue, be it regarding lack of time, money or expertise. Also, among the negative statements were worries about the capability of the available protocols to produce the information needed by the individual scientist. Phenotyping was predicted to become much more widespread in the future and its success was expected to depend on the development of reliable, fast and inexpensive methods. The study identified different aims of phenotyping and the suitability of the published protocols for these purposes was discussed. The contradiction between the limited use of characterization and its advantages was also discussed and proposals for the improvement of future phenotyping strategies are formulated.