Estimating mouse and rat use in American laboratories by extrapolation from Animal Welfare Act-regulated species.

Alone among Western nations, the United States has a two-tier system for welfare protections for vertebrate animals in research. Because its Animal Welfare Act (AWA) excludes laboratory rats and mice (RM), government veterinarians do not inspect RM laboratories and RM numbers are only partially reported to government agencies1. Without transparent statistics, it is impossible to track efforts to reduce or replace these sentient animals' use or to project government resources needed if AWA coverage were expanded to include them. I obtained annual RM usage data from 16 large American institutions and compared RM numbers to institutions' legally-required reports of their AWA-covered mammals. RM comprised approximately 99.3% of mammals at these representative institutions. Extrapolating from 780,070 AWA-covered mammals in 2017-18, I estimate that 111.5 million rats and mice were used per year in this period. If the same proportion of RM undergo painful procedures as are publicly reported for AWA-covered animals, then some 44.5 million mice and rats underwent potentially painful experiments. These data inform the questions of whether the AWA needs an update to cover RM, or whether the NIH should increase transparency of funded animal research. These figures can benchmark progress in reducing animal numbers in general and more specifically, in painful experiments. This estimate is higher than any others available, reflecting the challenges of obtaining statistics without consistent and transparent institutional reports.

Quality of interventional animal experiments in Chinese journals: compliance with ARRIVE guidelines.

BACKGROUND: In view of the inadequacy and incompleteness of currently-reported animal experiments and their overall poor quality, we retrospectively evaluated the reporting quality of animal experiments published in Chinese journals adhering to the Animal Research: Reporting of In Vivo Experiments (ARRIVE) guidelines. RESULTS: The databases CNKI, WanFang, VIP, and CBM were searched from inception until July 2018. Two appropriately-trained reviewers screened and extracted articles independently. The ARRIVE guidelines were used to assess the quality of the published reports of animal experiments. The compliance rate of every item was analyzed relative to their date of publication. A total of 4342 studies were included, of which 73.0% had been cited ≤5 times. Only 29.0% (1261/4342) were published in journals listed in the Chinese Science Citation Database. The results indicate that the compliance rate of approximately half of the sub-items (51.3%, 20/39) was less than 50%, of which 65.0% (13/20) was even less than 10%. CONCLUSIONS: The reporting quality of animal experiments in Chinese journals is not at a high level. Following publication of the ARRIVE guidelines in 2010, the compliance rate of the majority of its requirements has improved to some extent. However, less attention has been paid to the ethics and welfare of experimental animals, and a number of specific items in the Methods, Results, and Discussion sections continue to not be reported in sufficient detail. Therefore, it is necessary to popularize the ARRIVE guidelines, advocate researchers to adhere to them in the future, and in particular promote the use of the guidelines in specialized journals in order that the design, implementation, and reporting of animal experiments is promoted, to ultimately improve their quality.

Quality and validity of large animal experiments in stroke: A systematic review.

An important factor for successful translational stroke research is study quality. Low-quality studies are at risk of biased results and effect overestimation, as has been intensely discussed for small animal stroke research. However, little is known about the methodological rigor and quality in large animal stroke models, which are becoming more frequently used in the field. Based on research in two databases, this systematic review surveys and analyses the methodological quality in large animal stroke research. Quality analysis was based on the Stroke Therapy Academic Industry Roundtable and the Animals in Research: Reporting In Vivo Experiments guidelines. Our analysis revealed that large animal models are utilized with similar shortcomings as small animal models. Moreover, translational benefits of large animal models may be limited due to lacking implementation of important quality criteria such as randomization, allocation concealment, and blinded assessment of outcome. On the other hand, an increase of study quality over time and a positive correlation between study quality and journal impact factor were identified. Based on the obtained findings, we derive recommendations for optimal study planning, conducting, and data analysis/reporting when using large animal stroke models to fully benefit from the translational advantages offered by these models.

A semi-structured questionnaire survey of laboratory animal rehoming practice across 41 UK animal research facilities.

If a laboratory animal survives an experiment without lasting compromised welfare, its future must be negotiated. Rehoming may be a consideration. This paper reports on research findings that provide an indication of the uptake of animal rehoming by UK facilities and the associated moral, ethical, practical and regulatory considerations that inform decisions to rehome or not. This research addresses a widely acknowledged gap in the literature to understand both the numbers, and types of animals rehomed from UK research facilities, as well as the main motivations for engaging in the practice, and the barriers for those facilities not currently rehoming. From the ~160 UK research facilities in the UK, 41 facilities completed the questionnaire, giving a response rate of approximately 25%. Results suggest rehoming occurs routinely, yet the numbers are small; just 2322 animals are known to have been rehomed between 2015-2017. At least 1 in 10 facilities are rehoming. There exists a clear preference for the rehoming of some species (mainly cats, dogs and horses) over others (rodents, agricultural animals and primates). Indeed, although 94.15% of species kept in laboratories are rodents, they make up under a fifth (19.14%) of all animals known to be rehomed between 2015-2017. The primary motivation for rehoming is to boost staff morale and promote a positive ethical profile for the facility. Barriers include concern for the animal's welfare following rehoming, high scientific demand for animals that leaves few to be rehomed, and, finally, certain animals (mainly those genetically modified) are simply unsuited to rehoming. The findings of this research will support facilities choosing to rehome, as well as those that are not currently engaging in the practice. By promoting the practice, the benefits to rehoming in terms of improving laboratory animal's quality of life, helping facility staff to overcome the moral stress of killing, and addressing public concern regarding the fate of laboratory animals, can be attained. It is only once an understanding of rehoming from the perspective of UK research facilities has been ascertained, that appropriate policy and support can be provided.

Are the Lives of Animals Well-spent in Laboratory Science Research? A Study of Orthopaedic Animal Studies in Turkey.

BACKGROUND: As in all fields of medicine, animal studies are widely performed in orthopaedics and have increased in number over time. However, it is not clear to what extent these studies provide a basis for future research or advancements in clinical science. Concerns about the reliability and translational ability of animal studies have been reported, and major orthopaedic journals and organizations are encouraging the reduction of unnecessary experiments on animals. QUESTION/PURPOSES: (1) What proportion of animal studies conducted for orthopaedic research in Turkey were never published? And of those that were published, how long did it take to publish? (2) What proportion of those studies were published in journals with an Impact Factor of 2 or more? (3) What proportion of those published papers were never cited or cited only once? (4) What was the contribution to science of an animal euthanized for orthopaedic research in Turkey? METHODS: We reviewed all oral and poster presentations at the Turkish National Congress of Orthopaedics and Traumatology from 2009 to 2017 (retrieved from the archives of Acta Orthopaedica et Traumatologica Turcica), as well as all postgraduate theses in orthopaedics from 1991 to 2017 (retrieved from the archives of the National Thesis Center of the Council of Higher Education) to identify all orthopaedic studies that involved animals. We searched the keywords "animal studies," "experimental studies," and "orthopaedics" in these archives. We defined animal research as orthopaedic studies based on animal models. From this search and using that definition, 252 studies were identified. Of those, 4% (9) were excluded as they were thesis studies with no abstract in the archives. Thus, a total of 243 animal studies performed in Turkey were included for analysis in this retrospective study. The abstracts of these studies were examined to determine the study model (such as bone fracture models, tendon healing models, cartilage models) and number of euthanized animals. Between 1991 and 2017, 9412 vertebrate animals were euthanized for these studies. We searched PubMed, Google Scholar, ResearchGate, and ORCID to determine whether these papers were subsequently published, in which journal, and how long after the initial presentation publication occurred. The Web of Science 2019 database was used to determine the Impact Factor of the journals, the total citation count of each study, and the mean annual citation for each study (citations per year). For purposes of this analysis, we divided journals into those with an Impact Factor of 2 or more, 4 or more, and those with an Impact Factor below 2. The mean annual citation per euthanized animal (citations per animal per year) was calculated to determine the contribution of a euthanized animal to science. RESULTS: A total of 42% (101 of 243) of the animal studies in Turkey were never published. For all published studies, the mean time to publication was 2.2 ± 2.6 years (95% CI 1.7 to 2.6). The proportion of studies published in orthopaedic journals with an Impact Factor of 2 or more was 14% (34 of 243). Among the 142 published papers, 38% (54) were either never cited or were cited only once, and the mean citations per year was 1.1 ± 1.7 (95% CI 0.7 to 1.3). The mean citations per animal/year among the 142 published studies was 0.03 ± 0.04 (95% CI 0.02 to 0.04). CONCLUSION: In the 243 theses and national congress presentations, 9412 animals were euthanized. Based on the low percentage of papers using animals that were euthanized and the very low proportion of studies published in higher-Impact Factor journals or garnering more than a single citation, in aggregate, little seems to have been gained from the loss of animal life. Future studies should try to replicate or refute our results in other countries. CLINICAL RELEVANCE: Orthopaedic researchers should try to reduce their use of unnecessary animal studies, for example, by reporting on the use of the "3Rs" (replacement, reduction, and refinement) in the development of an animal study design, as well as through following universal guidelines so that a study might have a clinical impact. Researchers should not conduct an animal study until they are convinced that the expected results are quite likely to deliver substantial benefit to people or to advance science in a meaningful way; although this seems intuitive, our results suggest that this may not be taking place. Ethics committees in Turkey should consider more detailed questioning before approving animal studies. If our results are replicated elsewhere, then a broader look at how these approvals are conducted should be performed.

Factors Related to Animal Neglect in Brazilian Laboratories.

Our objectives were to identify the prevalence of negligence of laboratory animals in Brazil, to determine the primary factors associated with its occurrence and to suggest prevention strategies. A questionnaire was made available online between October 2015 and March 2016. A total of 116 respondents with experience in the use of laboratory animals and/or the use of alternative methods answered the questionnaire. Most respondents were women (77 respondents, 66.4%), a significant proportion had a degree in Veterinary Medicine (31 respondents, 27.2%), and a majority used animals in their work (88 respondents, 75.9%). Of the 88 animal users, 23 supplied information on the numbers and species of animals they used. When asked whether they knew that Brazilian law forbade animal experimentation when alternative methods exist, seven (9.1%) respondents mentioned Act 9605/1998. Most, but not all, respondents (96 respondents, 82.8%) submitted their projects to an Animal Use and Ethics Committee (AUEC), and many (65 respondents, 56%) reported their belief that animal neglect occurred at their institution. Negligence was found to be associated with: institutions where the numbers of animals used were not recorded (p = 0.008); institutions where respondents were unaware of the relevant legislation, that is, Act 9605/1998 (p = 0.042); or where there was evidence that not all project proposals were submitted to the AUEC or evidence of no submissions at all (p = 0.022). Negligence of animals was found to be highly prevalent. Prevention strategies might involve increased transparency to the general public, the empowerment of individuals that work with animals to report any concerns, optimised inspection of facilities where animal work is carried out and significant improvements to the role of AUECs.

Publication and Citation Analysis of Medical Doctors' Residency Master's Theses Involving Animal Experiments on Rats in Turkey.

The number of non-human animals used in research has increased in line with advances in medical technology, although it has previously been shown that these experiments demonstrate poor human utility. This study aimed to determine the effectiveness of animal studies on rats that were performed as part of medical doctors' residency master's theses prepared in Turkey between January 2006 and December 2015. The number of thesis-derived published papers from each year, as well as the subsequent citation rate of these papers, was determined. Results from 34% of the 656 analysed studies (226/656) were published as papers in PubMed-indexed journals. These 226 studies got 1803 subsequent citations in total. Citation counts were statistically significantly different in 2009 and 2010, as compared to 2011, 2013, 2014 and 2015. Previous studies showed that the usual main objective for carrying out animal studies in Turkey was the preparation of a thesis or the furthering of an academic career (i.e. personal self-interest). In the current study, the publication rate and the number of subsequent citations of these thesis-derived papers were both low, and thus, the contribution of these animal studies to scientific progress is doubtful. It is recommended that institutional research ethics committees should be much more highly selective in approving the use of animals for the purposes of student thesis preparation.

Attitudes towards animal study registries and their characteristics: An online survey of three cohorts of animal researchers.

OBJECTIVES: Prospective registration of animal studies has been suggested as a new measure to increase value and reduce waste in biomedical research. We sought to further explore and quantify animal researchers' attitudes and preferences regarding animal study registries (ASRs). DESIGN: Cross-sectional online survey. SETTING AND PARTICIPANTS: We conducted a survey with three different samples representing animal researchers: i) corresponding authors from journals with high Eigenfactor, ii) a random Pubmed sample and iii) members of the CAMARADES network. MAIN OUTCOME MEASURES: Perceived level of importance of different aspects of publication bias, the effect of ASRs on different aspects of research as well as the importance of different research types for being registered. RESULTS: The survey yielded responses from 413 animal researchers (response rate 7%). The respondents indicated, that some aspects of ASRs can increase administrative burden but could be outweighed by other aspects decreasing this burden. Animal researchers found it more important to register studies that involved animal species with higher levels of cognitive capabilities. The time frame for making registry entries publicly available revealed a strong heterogeneity among respondents, with the largest proportion voting for "access only after consent by the principal investigator" and the second largest proportion voting for "access immediately after registration". CONCLUSIONS: The fact that the more senior and experienced animal researchers participating in this survey clearly indicated the practical importance of publication bias and the importance of ASRs underscores the problem awareness across animal researchers and the willingness to actively engage in study registration if effective safeguards for the potential weaknesses of ASRs are put into place. To overcome the first-mover dilemma international consensus statements on how to deal with prospective registration of animal studies might be necessary for all relevant stakeholder groups including animal researchers, academic institutions, private companies, funders, regulatory agencies, and journals.

Publication rates in animal research. Extent and characteristics of published and non-published animal studies followed up at two German university medical centres.

Non-publication and publication bias in animal research is a core topic in current debates on the "reproducibility crisis" and "failure rates in clinical research". To date, however, we lack reliable evidence on the extent of non-publication in animal research. We collected a random and stratified sample (n = 210) from all archived animal study protocols of two major German UMCs (university medical centres) and tracked their results publication. The overall publication rate was 67%. Excluding doctoral theses as results publications, the publication rate decreased to 58%. We did not find substantial differences in publication rates with regard to i) the year of animal study approval, ii) the two UMCs, iii) the animal type (rodents vs. non-rodents), iv) the scope of research (basic vs. preclinical), or v) the discipline of the applicant. Via the most reliable assessment strategy currently available, our study confirms that the non-publication of results from animal studies conducted at UMCs is relatively common. The non-publication of 33% of all animal studies is problematic for the following reasons: A) the primary legitimation of animal research, which is the intended knowledge gain for the wider scientific community, B) the waste of public resources, C) the unnecessary repetition of animal studies, and D) incomplete and potentially biased preclinical evidence for decision making on launching early human trials. Results dissemination should become a professional standard for animal research. Academic institutions and research funders should develop effective policies in this regard.

Experimental Animal Use in Turkey: A Comparison with Other Countries.

The legal structure that governs animal use in Turkey is in line with that of the European Union (EU). In 2004, legislation on the use of animals for experimental and other scientific purposes was established in Turkey for the first time. The present study aimed to compare the data on experimental animal use in Turkey (during the period 2008-2017) with similar reports from selected countries (the United States, Australia, Canada and the EU). In Turkey, a total of 2,104,828 animals were used for experimental and other scientific purposes during 2008-2017. Of the animals used, 758,887 were fish (36%), 433,417 rats (21%), 302,512 birds other than quail (14%) and 285,531 mice (13%). According to a breakdown by purpose for use, in Turkey during 2009-2017, out of a total number of 1,955,307 animals used, 56% were for fundamental biological studies, with a high proportion used for research on animal disease. Compared with the other countries, fewer animals were used in Turkey although the national trend seems to indicate that the number is fluctuating. Further studies are required to uncover the reasons behind this reduced animal use in Turkey, as compared to other countries.

Reporting and analysis of repeated measurements in preclinical animals experiments.

A common feature of preclinical animal experiments is repeated measurement of the outcome, e.g., body weight measured in mice pups weekly for 20 weeks. Separate time point analysis or repeated measures analysis approaches can be used to analyze such data. Each approach requires assumptions about the underlying data and violations of these assumptions have implications for estimation of precision, and type I and type II error rates. Given the ethical responsibilities to maximize valid results obtained from animals used in research, our objective was to evaluate approaches to reporting repeated measures design used by investigators and to assess how assumptions about variation in the outcome over time impact type I and II error rates and precision of estimates. We assessed the reporting of repeated measures designs of 58 studies in preclinical animal experiments. We used simulation modelling to evaluate three approaches to statistical analysis of repeated measurement data. In particular, we assessed the impact of (a) repeated measure analysis assuming that the outcome had non-constant variation at all time points (heterogeneous variance) (b) repeated measure analysis assuming constant variation in the outcome (homogeneous variance), (c) separate ANOVA at individual time point in repeated measures designs. The evaluation of the three model fitting was based on comparing the p-values distributions, the type I and type II error rates and by implication, the shrinkage or inflation of standard error estimates from 1000 simulated dataset. Of 58 studies with repeated measures design, three provided a rationale for repeated measurement and 23 studies reported using a repeated-measures analysis approach. Of the 35 studies that did not use repeated-measures analysis, fourteen studies used only two time points to calculate weight change which potentially means collected data was not fully utilized. Other studies reported only select time points (n = 12) raising the issue of selective reporting. Simulation studies showed that an incorrect assumption about the variance structure resulted in modified error rates and precision estimates. The reporting of the validity of assumptions for repeated measurement data is very poor. The homogeneous variation assumption, which is often invalid for body weight measurements, should be confirmed prior to conducting the repeated-measures analysis using homogeneous covariance structure and adjusting the analysis using corrections or model specifications if this is not met.

Conscientious Objection to Animal Testing: A Preliminary Survey Among Italian Medical and Veterinary Students.

The use of animals for educational and research purposes is common in both veterinary and human medicine degree courses, and one that involves important ethical considerations. The aim of this study was to assess the extent of differences between the knowledge and attitudes of veterinary students and medical students on animal bioethics, on alternative strategies and on their right to conscientiously object to animal experimentation. To this end, a questionnaire was completed by 733 students (384 human medicine students (HMS) and 349 veterinary medicine students (VMS)). VMS were more aware than HMS (72.2% and 59.6%, respectively) of the existence of an Italian law on the right to conscientiously object to animal experimentation. However, very few of them had exercised this right. Many VMS (43.3%) felt that animal bioethics courses should be mandatory (only 17.4% of HMS felt the same way). More VMS than HMS (81.7% and 59.1%, respectively) expressed an interest in attending a course on alternatives to animal experimentation. The data suggest the need for appropriate educational interventions, in order to allow students to make choices based on ethical principles. Fostering close collaborations between departments of human medicine and veterinary medicine, for example, through shared study modules, could promote the development of ethical competence as a basic skill of students of both veterinary and human medicine courses.

Experiments done in Black-6 mice: what does it mean?

Low replicability of animal experiments is perceived as a major hurdle in the field of biomedicine. Attempts to enhance the replicability and to reduce the variability in basic research has led to the recommendation to use isogenic mice. The C57BL/6 strain has evolved as a gold standard strain for this purpose. However, C57BL/6 mice are maintained as substrains by multiple vendors. Evidence exists that the subtle differences between these mouse lines have not been systematically investigated and are often ignored. In the present study, we characterized the female mice of two closely related substrains (C57BL/6J and C57BL/6N) from three vendors in Europe (Charles River Laboratories, Envigo, Janvier Labs) in a battery of behavioral tests. Our data show and confirm substantial behavioral differences between the C57BL/6J and C57BL/6N mice. Importantly, the substrain differences were largely affected by the origin of the animals, as a significant effect of vendor or interaction between the substrain and vendor occurred in all tests. This work highlights the importance of adhering to precise international nomenclature in all publications reporting animal experiments. Moreover, the generalization of research findings from a single mouse substrain can be seriously limited due to genetic drift and environmental variables occurring at different vendors. However, heterogenization of samples, by including animals of different substrains, can enhance generalizability. These issues need to be seriously addressed to improve reproducibility, replicability, and the translational potential of the mouse models.

An Estimate of the Number of Animals Used for Scientific Purposes Worldwide in 2015.

Few attempts have been made to estimate the global use of animals in experiments, since our own estimated figure of 115.2 million animals for the year 2005. Here, we provide an update for the year 2015. Data from 37 countries that publish national statistics were standardised against the definitions of 'animals' and 'procedures' used in the European Union (EU) Directive 2010/63/EU. We also applied a prediction model, based on publication rates, to estimate animal use in a further 142 countries. This yielded an overall estimate of global animal use in scientific procedures of 79.9 million animals, a 36.9% increase on the equivalent estimated figure for 2005, of 58.3 million animals. We further extrapolated this estimate to obtain a more comprehensive final global figure for the number of animals used for scientific purposes in 2015, of 192.1 million. This figure included animals killed for their tissues, normal and genetically modified (GM) animals without a harmful genetic mutation that are used to maintain GM strains and animals bred for laboratory use but not used. Since the 2005 study, there has been no evident increase in the number of countries publishing data on the numbers of animals used in experiments. Without regular, accurate statistics, the impact of efforts to replace, reduce and refine animal experiments cannot be effectively monitored.

Guidelines on statistics for researchers using laboratory animals: the essentials.

There is growing concern that the omnipresence of flawed statistics and the deficient reproducibility that arises therefrom results in an unethical waste of animals in research. The present review aims at providing guidelines in biostatistics for researchers, based on observed frequent mistakes, misuses and misconceptions as well as on the specificities of animal experimentation. Twelve recommendations are formulated that cover sampling, sample size optimisation, choice of statistical tests, understanding p-values and reporting results. The objective is to expose important statistical issues that one should consider for the correct design, execution and reporting of experiments.