Reichlin TS, Vogt L and Würbel H (2016). Oxford University Press. Where available, cite the Research Resource Identifier (RRID) for reagents and tools used [7,8]. Evaluation of excess significance bias in animal studies of neurological diseases. Distinguishing between exploratory and confirmatory preclinical research will improve translation. Examples or references about Soc 1 Report Example and Applying the Arrive Guidelines to An In Vivo Database that we get come from reputable online resources. Hypothesis-testing studies also have a pre-defined primary outcome measure, which is used to assess the evidence in support of the specific research question (see item 6 – Outcome measures). Power Analysis Software for Educational Researchers. Thoughtful consideration is warranted, as additional unnecessary animal studies are wasteful and unethical. Provide details of housing and husbandry conditions, including any environmental enrichment. The Assessment of Science: The Relative Merits of Post-Publication Review, the Impact Factor, and the Number of Citations. Data were analysed as an REML using the MIXED procedure of SAS. 0000178764 00000 n Therefore, we used intraperitoneal injection of 18F-FDG for our experiments evaluating the influence of animal handling on 18F-FDG biodistribution.” [1], “Since Xenopus oocytes have a higher potential for homologous recombination than fertilized embryos…we next tested whether the host transfer method could be used for efficient HDR-mediated knock-in. Bero L, Anglemyer A, Vesterinen H and Krauth D (2016). cage location, day or week of procedure). Request pocket-sized ARRIVE guidelines P ocket-sized versions of the ARRIVE guidelines 2.0 (English only) allow quick reference to the guidelines and dissemination in a handy, easy-to-use format. If none exist, this should be stated. This also includes the potential exploitation of the scientific advance being reported for the institution, the authors, or the research funders. Data should follow the FAIR guiding principles [6], that is data are findable, accessible (i.e. R Foundation for Statistical Computing,. d. Francisella tularensis: No Evidence for Transovarial Transmission in the Tularemia Tick Vectors Dermacentor reticulatus and Ixodes ricinus. Available at: Bustamante R, Daza MA, Canfrán S, García P, Suárez M, Trobo I and Gómez de Segura IA (2018). Significant lymphoproliferation could be seen only in the murine pre-T cells. However, abstracts often either do not contain enough information for this purpose [2], or contain information that is inconsistent with the results in the rest of the manuscript [3,4]. age, sex, strain, species), the intervention (e.g. Fourth Edition. Akkers RC, van Heeringen SJ, Jacobi UG, Janssen-Megens EM, Francoijs KJ, Stunnenberg HG and Veenstra GJ (2009). “Blood pressure, heart rate, oxygen saturation and amount of blood extracted were recorded every 5 minutes. Effects of increasing supplemental dietary Zn concentration on growth performance and carcass characteristics in finishing steers fed ractopamine hydrochloride. The effects of routine cage-changing on cardiovascular and behavioral parameters in male Sprague-Dawley rats. This includes ensuring that they have the relevant approval for their study from an appropriate ethics committee and/or regulatory body before the work starts. Pain and Laboratory Animals: Publication Practices for Better Data Reproducibility and Better Animal Welfare. Emery M, Nanchen N, Preitner F, Ibberson M and Roduit R (2016). initial tumour volume)Â, Animal characteristics (e.g. sex, age bracket, weight bracket)Â, Location – exposure to light, ventilation and disturbances may vary in cages located at different height or on different racks, which may affect important physiological processes. Animal Welfare Standards and International Collaborations. Specific details to report include, but are not limited to, structural enrichment (e.g. The species, strain, substrain, sex, weight, and age of animals are critical factors that can influence most experimental results [1-5]. Allocation concealment refers to concealing the treatment to be allocated to each individual animal from those assigning the animals to groups, until the time of assignment. Mamrot J, Legaie R, Ellery SJ, Wilson T, Seemann T, Powell DR, Gardner DK, Walker DW, Temple-Smith P, Papenfuss AT and Dickinson H (2017). In a randomised controlled trial, manuscripts reporting in vivo animal research submitted to PLOS ONE (March–June 2015) were … development of motor impairments that could affect behavioural measurements). 0000008426 00000 n Minimisation is an alternative strategy to allocate animals/samples to treatment group to balance variables that might influence the result of an experiment. The alternative hypothesis (H1) postulates that an effect exists.Â, Outcome measure: Any variable recorded during a study to assess the effects of a treatment or experimental intervention. (Access Date: 10 january 2020). Together with randomisation, allocation concealment helps minimise selection bias, which can introduce systematic differences between treatment groups. Nuisance variables increase variability in the sample, which reduces statistical power. RO���rˡxp��K�. An experiment with a directional (one-sided) H1 can be powered and analysed with a one-sided test with the goal of maximising the sensitivity to detect this directional effect. We set out to analyze the fragile-skin phenotype in rab-6.2(ok2254) animals genetically…We observed several ruptured animals on our rab-6.2(ok2254) culture plates during normal maintenance, a phenotype very rarely observed in wild-type cultures…We hypothesized that RAB-6.2 is required for skin integrity.” [3]. Inferential statistics are used to make generalisations about the population from which the samples are drawn. 0000006658 00000 n Provide further relevant information on the provenance of animals, health/immune status, genetic modification status, genotype, and any previous procedures. ), and technical considerations (cage, time of day, batch, experimenter, etc. It is important to interpret the results of the study in the context of the study objectives (see item 13 – Objectives). two-tailed student’s t-test with a 0.05 significance threshold)Â, the effect size of interest and a justification explaining why an effect size of that magnitude is relevantÂ, the estimate of variability used (e.g. This relationship has to be managed in the statistical analysis by including cage/litter/animal effects in the model or by aggregating the outcome measure to the cage/litter/animal level. Thus, describing the reality of the experiment and the hierarchy of the data, along with the measures taken in the design and the analysis to account for this hierarchy, is crucial to assessing whether the statistical methods used are appropriate. The ARRIVE (Animal Research: Reporting of In Vivo Experiments) guidelines were developed to improve standards of reporting the results of animal experiments. Kallnik M, Elvert R, Ehrhardt N, Kissling D, Mahabir E, Welzl G, Faus-Kessler T, de Angelis MH, Wurst W, Schmidt J and Holter SM (2007). body weights must be within a certain range for a particular procedure) or related to other study parameters (e.g. Husbandry of zebrafish, Danio rerio, and the cortisol stress response. Ruxton G and Colegrave N (2017). Also indicate the total number of animals used. To make the variation more uniform, we used a logarithmic transform of each fish's cortisol per unit weight as the dependent variable in our analyses. Similarly, human clinical trials initiated based on insufficient or misleading animal research evidence increase research waste and negatively influence the risk-benefit balance for research participants [1,2]. The examples given are from a wide range of research using a range of animal species. Sundberg JP and Schofield PN (2010). We tested for the effect of the following husbandry and environmental risk factors: cage mate relationships (i.e. Blinding is a strategy used to minimise these subjective biases. For example: a. R: A language and environment for statistical computing. Federer LM, Lu Y-L, Joubert DJ, Welsh J and Brandys B (2015). Leaving out of the abstract information such as the species of animal used or the drugs being tested, limits the value of preclinical systematic reviews as relevant studies cannot be identified and included. A guide to defining and implementing protocols for the welfare assessment of laboratory animals: eleventh report of the BVAAWF/FRAME/RSPCA/UFAW Joint Working Group on Refinement. For complex study designs, a visual representation is more easily interpreted than a text description, so a timeline diagram or flow chart is recommended. Any signs of illness were logged as “adverse events” in the experiment, the mouse was immediately killed, and an autopsy was performed to establish the cause of illness.” [1], “Although procedures were based on those reported in the literature, dogs under Protocol 1 displayed high levels of stress and many experienced vomiting. Enrichment given to pigs housed in isolation had no effect on plasma cortisol, but greatly reduced it in non-isolated pigs. British Ecological Society (2017). Report the health status of all animals used in the study, and any previous procedures the animals have undergone. Categorical data can be expressed as counts, frequencies, or proportions. Polo J, Rodríguez C, Ródenas J, Russell LE, Campbell JM, Crenshaw JD, Torrallardona D and Pujols J (2015). Commentary: Mouse Genetic Nomenclature:Standardization of Strain, Gene, and Protein Symbols. Beliefs or activism (e.g. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.” [3], Bias: The over- or under-estimation of the true effect of an intervention. Bias is caused by inadequacies in the design, conduct, or analysis of an experiment, resulting in the introduction of error.Â, Descriptive and inferential statistics: Descriptive statistics are used to summarise the data. The environment determines the health and wellbeing of the animals and every aspect of it can potentially affect their behavioural and physiological responses, thereby affecting research outcomes [1]. Consequently, low powered studies contribute to the poor internal validity of research and risk wasting animals used in inconclusive research [6]. A manifesto for reproducible science. Focusing on the Essential 10 in priority better allows journal staff, editors and reviewers to verify that the items have been adequately reported in manuscripts. Pigs were catheterised and placed into the PigTurns® 48 h post recovery. Bate ST and Clark RA (2014). Conour LA, Murray KA and Brown MJ (2006). These practices, however transparent, still need to be accompanied by a disclosure on the reasoning for data cleaning, and whether methods were defined before any data were collected. Gouveia K and Hurst JL (2017). The one-chamber social interaction test requires manual scoring and was analyzed by an individual with no knowledge of the questions.” [3]. Welfare or safety reasons may prevent blinding of animal care staff but in most cases, blinding is possible. If no power calculation was used to determine the sample size, state this explicitly and provide the reasoning that was used to decide on the sample size per group. Regardless of whether a power calculation was used or not, when explaining how the sample size was determined take into consideration any anticipated loss of animals or data, for example due to exclusion criteria established upfront or expected attrition (see item 3 – Inclusion and exclusion criteria). Secondary outcomes constitute Rotarod performance, stroke volume (quantified on MR imaging or brain sections, respectively), diffusion tensor imaging (DTI) connectome mapping, and histological analyses to measure neuronal and microglial densities, and phagocytic activity…The study is designed with 80% power to detect a relative 25% difference in pellet-reaching performance in the Staircase test.” [1]Â, “The primary endpoint of this study was defined as left ventricular ejection fraction (EF) at the end of follow-up, measured by magnetic resonance imaging (MRI). Independent variables can be scientifically interesting, or nuisance variables. Evidence for preclinical research also indicates that funding sources may influence the interpretation of study outcomes [4,5]. It is especially important to use randomisation in situations where it is not possible to blind all or parts of the experiment but even with randomisation, researcher bias can pervert the allocation. Report all analyses performed in full. Relevant information to describe the statistical methods include:Â. larkson JM, Dwyer DM, Flecknell PA, Leach MC and Rowe C (2018). Provide the name of the ethical review committee or equivalent that has approved the use of animals in this study and any relevant licence or protocol numbers (if applicable). Effects of MVA85A vaccine on tuberculosis challenge in animals: systematic review. There are several types of studies where a priori sample size calculations are not appropriate. Comment on whether, and how, the findings of this study are likely to generalise to other species or experimental conditions, including any relevance to human biology (where appropriate). For each analysis, report the exact value of n in each experimental group. Ideally, investigators should be unaware of the treatment(s) animals have received or will be receiving, from the start of the experiment until the data have been analysed. To address this problem we are hosting a SequenceServer32 BLAST-search website (http://spinymouse.erc.monash.edu/sequenceserver/http://spinymouse.erc.monash.edu/sequenceserver/). Explanation and Elaboration document for the ARRIVE guidelines. On arrival from El-Nile Company, animals were assigned a group designation and weighed. Neurodegeneration in an animal model of Parkinson's disease is exacerbated by a high-fat diet. A new behavioural apparatus to reduce animal numbers in multiple types of spontaneous object recognition paradigms in rats. Cohorts of at least seven homozygote mice of each sex per pipeline were generated…The random allocation of mice to experimental group (wildtype versus knockout) was driven by Mendelian Inheritance.” [3]Â. Membership of a relevant advocacy or lobbying organisation. The combination of no enrichment and isolation maximally impacted tear staining and eosinophil numbers. The animals were excluded if insertion of the thread resulted in perforation of the vessel wall (determined by the presence of sub-arachnoid blood at the time of sacrifice), if the silicon tip of the thread became dislodged during withdrawal, or if the animal died prematurely, preventing the collection of behavioral and histological data.” [1]. The Researchers’ View of Scientific Rigor—Survey on the Conduct and Reporting of In Vivo Research. Tank colour affects stress in aquatic species [5,6]. In the 10 years since publication, the ARRIVE guidelines have been endorsed by more than a thousand journals from across the life sciences. Stage 1 Registered Report: Effect of deficient phagocytosis on neuronal survival and neurological outcome after temporary middle cerebral artery occlusion (tMCAo) [version 3; referees: 2 approved]. 0000233164 00000 n Estrous cycle staging before mating led to increased efficiency in the production of pseudopregnant recipients without negatively affecting embryo transfer in mice. (2016). 2010. Friends, family, relationships, and other close personal connections to people who may potentially benefit financially or in other ways from the research. the nuisance variables taken into account in each statistical test (e.g. as blocking factors or covariates), Â, what statistical analyses were performed and references for the methods used Â, the software package and version used, including computer code if availableÂ. ; see “Considerations for the randomisation strategy” below), the method used to generate the randomisation sequence (e.g. Whole Animal Experiments Should Be More Like Human Randomized Controlled Trials. Discuss the range of circumstances in which the effect is observed, and factors which may moderate that effect. Ruxton GD and Neuhäuser M (2010). Rosenthal R and Fode KL (1963). Effect of diet and animal care/housing protocols on body weight, survival, tumor incidences, and nephropathy severity of F344 rats in chronic studies. 0000080208 00000 n The protein concentration was determined with the Pierce BCA assay kit (catalogue number 23225; Thermo Fisher Scientific, Rockford, IL, USA). This led us to significantly alter procedures in order to optimize the protocol for the purposes of our own fasting and postprandial metabolic studies.” [2]. A guide to reproducible code in ecology and evolution. Learn collocations of Arrive with free vocabulary lessons. Relationship between Sponsorship and Failure Rate of Dental Implants: A Systematic Approach. Box plots show the 25%, 50% and 75% percentiles; whiskers show 10% and 90% percentile. batch of reagent or experimenter), or as a covariate (e.g. Reporting standards for preclinical studies of stroke therapy. For each outcome measure, indicate numbers clearly within the text or on figures, and provide absolute numbers (e.g. Treatment allocation in controlled trials: why randomise? 0000014292 00000 n One entire pen (ZnAA90) was removed as an outlier from both Pre-RAC and RAC periods for poor performance caused by illness unrelated to treatment...Outliers were determined using Cook’s D statistic and removed if Cook’s D > 0.5. Comment on the study limitations including potential sources of bias, limitations of the animal model, and imprecision associated with the results. Provide a clear statement explaining how the study conforms to appropriate regulations and guidelines. For hypothesis-testing experiments, where inferential statistics are used to estimate the size of the effect and to determine the weight of evidence against the null hypothesis, the sample size needs to be justified to ensure experiments are of an optimal size to test the research question [1,2] (see item 13 – Objectives). Sample sizes that are too small (i.e. Pradillo JM, Denes A, Greenhalgh AD, Boutin H, Drake C, McColl BW, Barton E, Proctor SD, Russell JC, Rothwell NJ and Allan SM (2012). Specify how the outcome measure(s) assessed are relevant to the objectives of the study.Â, “The following parameters were assessed: threshold pressure (TP; intravesical pressure immediately before micturition); post-void pressure (PVP; intravesical pressure immediately after micturition); peak pressure (PP; highest intravesical pressure during micturition); capacity (CP; volume of saline needed to induce the first micturition); compliance (CO; CP to TP ratio); frequency of voiding contractions (VC) and frequency of non-voiding contractions (NVCs).” [1]Â. Delayed kindling epileptogenesis and increased neurogenesis in adult rats housed in an enriched environment. Summary/descriptive statistics provide a quick and simple description of the data, they communicate quantitative results easily and facilitate visual presentation. If health status is unknown or was not tested, explicitly state this. intraperitoneal administration vs. oral gavage), or animals that are used as their own control in a longitudinal study. Limitations are unavoidable in scientific research, and describing them is essential to share experience, guide best practice, and aid the design of future experiments [2]. If the study did not have humane endpoints, state this. Guidance for the description of animal research in scientific publications. Hannan AJ (2014). The use of humane endpoints can help minimise harm while allowing the scientific objectives to be achieved [1]. For example, the number of animals needed for antibody or tissue production is determined by the amount required and the production ability of an individual animal. ABSTRACT 2 Provide an accurate summary of the background, research objectives, including details of the species or strain of animal used, key methods, principal findings and conclusions of the study. Gaspani L, Bianchi M, Limiroli E, Panerai AE and Sacerdote P (2002). different drugs of the same class), and the outcome measured (e.g. Include any accreditation of laboratories if appropriate (e.g. Michel MC, Murphy TJ and Motulsky HJ (2020). The maximum caging density was five mice from the same litter and sex starting from weaning. Study design can influence the statistical power of an experiment and the power calculation used needs to be appropriate for the design implemented. Statistical programs to help perform a priori sample size calculations exist for a variety of experimental designs and statistical analyses, both freeware (web based applets and functions in R) and commercial software [7-9]. Choosing the appropriate calculator or algorithm to use depends on the type of outcome measures and independent variables, and the number of groups. Consultation with a statistician is recommended, especially when the experimental design is complex or unusual.

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