For each experimental group, including controls, describe what was done, how it was done, and what was used.
Explanation
Essential information to describe in the manuscript includes the procedures used to develop the model (e.g., induction of the pathology), the procedures used to measure the outcomes, and pre- and postexperimental procedures, including animal handling, welfare monitoring, and euthanasia. Animal handling can be a source of stress, and the specific method used (e.g., mice picked up by tail or in cupped hands) can affect research outcomes1–3. Details about animal care and monitoring intrinsic to the procedure are discussed in further detail in Item 16. Animal care and monitoring. Provide enough detail to enable others to replicate the methods and highlight any quality assurance and quality control used4,5. A schematic of the experimental procedures with a timeline can give a clear overview of how the study was conducted. Information relevant to distinct types of interventions and resources are described in Table 1.
Table 1: Examples of information to include when reporting specific types of experimental procedures and resources. AVMA, American Veterinary Medical Association; RRID, Research Resource Identifier.
Procedures
Resources
Pharmacological procedures (intervention and control)
Drug formulation
Dose
Volume
Concentration
Site and route of administration
Frequency of administration
Vehicle or carrier solution formulation and volume
Any evidence that the pharmacological agent used reaches the target tissue
Anaesthetic used (including dose and other information listed in pharmacological procedures section above)
Pre- and postanalgesia regimen
Presurgery procedures (e.g., fasting)
Aseptic techniques
Monitoring (e.g., assessment of surgical anaesthetic plane)
Whether the procedure is terminal or not
Postsurgery procedures
Duration of the procedure and duration of anaesthesia
Physical variables measured
Reagents (e.g., antibodies, chemicals)
Manufacturer
Supplier
Catalogue number
Lot number (if applicable)
Purity of the drug (if applicable)
RRID
Pathogen infection (intervention and control)
Infectious agent
Dose load
Vehicle or carrier solution formulation and volume
Site and route of infection
Timing or frequency of infection
Equipment and software
Manufacturer
Supplier
Model/version number
Calibration procedures (if applicable)
RRID
Euthanasia
Method of euthanasia, including the humane standards the method complies with, such as the AVMA8
Pharmacological agent, if used (including dose and information listed in pharmacological procedures section above)
Any measures taken to reduce pain and distress before or during euthanasia
Timing of euthanasia
Tissues collected post-euthanasia and timing of collection
When available, cite the Research Resource Identifier (RRID) for reagents and tools used6,7. RRIDs are unique and stable, allowing unambiguous identification of reagents or tools used in a study, aiding other researchers to replicate the methods.
Detailed step-by-step procedures can also be saved and shared online, for example, using Protocols.io9, which assigns a digital object identifier (DOI) to the protocol and allows cross-referencing between protocols and publications.
Examples
Example of how to report this item. This figure is an alternative version of the figure published in reference10
‘For the diet-induced obesity (DIO) model, eight-week-old male mice had ad libitum access to drinking water and were kept on standard chow (SFD, 10.9 kJ/g) or on western high-fat diet (HFD; 22 kJ/g; kcal from 42% fat, 43% from carbohydrates and 15% from protein; E15721-34, Ssniff, Soest, Germany) for 15 weeks (https://dx.doi.org/10.17504/protocols.io.kbacsie)’11.
‘The frozen kidney tissues were lysed. The protein concentration was determined with the Pierce BCA assay kit (catalogue number 23225; Thermo Fisher Scientific, Rockford, IL, USA). A total of 100–150 μg total proteins were resolved on a 6–12% SDS-PAGE gel. The proteins were then transferred to a nitrocellulose membrane, blocked with 5% skimmed milk for 1 h at room temperature and incubated overnight at 4°C with primary antibodies against the following proteins: proliferating cell nuclear antigen (PCNA; Cat# 2586, RRID: AB_2160343), phospho-AMPK (Cat# 2531, RRID: AB_330330), phospho-mTOR (Cat# 2971, RRID: AB_330970)…. The β-actin (Cat# A5441, RRID: AB_476744) antibody was obtained from Sigma. The blots were subsequently probed with HRP-conjugated anti-mouse (Cat# A0216) or anti-rabbit IgG (Cat# A0208; Beyotime Biotechnology, Beijing, China) at 1:1000. Immunoreactive bands were visualized by enhanced chemiluminescence, and densitometry was performed using ImageJ software (RRID: SCR_003070, Bio-Rad Laboratories)’12.
Training
The UK EQUATOR Centre runs training on how to write using reporting guidelines.
Discuss this item
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References
1.
Gouveia K, Hurst JL. Optimising reliability of mouse performance in behavioural testing: The major role of non-aversive handling. Scientific Reports. 2017;7(1). doi:10.1038/srep44999
2.
Clarkson JM, Dwyer DM, Flecknell PA, Leach MC, Rowe C. Handling method alters the hedonic value of reward in laboratory mice. Scientific Reports. 2018;8(1). doi:10.1038/s41598-018-20716-3
3.
Hurst JL, West RS. Taming anxiety in laboratory mice. Nature Methods. 2010;7(10):825-826. doi:10.1038/nmeth.1500
4.
Hewitt JA, Brown LL, Murphy SJ, Grieder F, Silberberg SD. Accelerating biomedical discoveries through rigor and transparency. ILAR Journal. 2017;58(1):115-128. doi:10.1093/ilar/ilx011
5.
Almeida JL, Cole KD, Plant AL. Standards for cell line authentication and beyond. PLOS Biology. 2016;14(6):e1002476. doi:10.1371/journal.pbio.1002476
6.
Bandrowski AE, Martone ME. RRIDs: A simple step toward improving reproducibility through rigor and transparency of experimental methods. Neuron. 2016;90(3):434-436. doi:10.1016/j.neuron.2016.04.030
7.
Bandrowski A, Brush M, Grethe JS, et al. The resource identification initiative: A cultural shift in publishing. Journal of Comparative Neurology. 2015;524(1):8-22. doi:10.1002/cne.23913
8.
Leary SL, underwood w, anthony r, cartner s, corey d, grandin t, et al. AVMA guidelines for the euthanasia of animals: 2013 edition. AVMA; 2013.
9.
Teytelman L. 2016;35.
10.
Reynolds P. Published online 2018.
11.
Bauters D, Bedossa P, Lijnen HR, Hemmeryckx B. Functional role of ADAMTS5 in adiposity and metabolic health. Souza-Mello V, ed. PLOS ONE. 2018;13(1):e0190595. doi:10.1371/journal.pone.0190595
12.
Lian X, Wu X, Li Z, et al. The combination of metformin and 2‐deoxyglucose significantly inhibits cyst formation in miniature pigs with polycystic kidney disease. British Journal of Pharmacology. 2019;176(5):711-724. doi:10.1111/bph.14558
Citation
For attribution, please cite this work as:
Sert
NP du, Hurst V, Ahluwalia A, et al. The ARRIVE reporting guideline for
writing animal research articles. The EQUATOR Network guideline
dissemination platform. doi:10.1234/equator/1010101
Reporting Guidelines are recommendations to help describe your work clearly
Your research will be used by people from different disciplines and backgrounds for decades to come. Reporting guidelines list the information you should describe so that everyone can understand, replicate, and synthesise your work.
Reporting guidelines do not prescribe how research should be designed or conducted. Rather, they help authors transparently describe what they did, why they did it, and what they found.
Reporting guidelines make writing research easier, and transparent research leads to better patient outcomes.
Easier writing
Following guidance makes writing easier and quicker.
You work will be read by different people, for different reasons, around the world, and for decades to come. Reporting guidelines help you consider all of your potential audiences. For example, your research may be read by researchers from different fields, by clinicians, patients, evidence synthesisers, peer reviewers, or editors. Your readers will need information to understand, to replicate, apply, appraise, synthesise, and use your work.
Cohort studies
A cohort study is an observational study in which a group of people with a particular exposure (e.g. a putative risk factor or protective factor) and a group of people without this exposure are followed over time. The outcomes of the people in the exposed group are compared to the outcomes of the people in the unexposed group to see if the exposure is associated with particular outcomes (e.g. getting cancer or length of life).
A case-control study is a research method used in healthcare to investigate potential risk factors for a specific disease. It involves comparing individuals who have been diagnosed with the disease (cases) to those who have not (controls). By analysing the differences between the two groups, researchers can identify factors that may contribute to the development of the disease.
An example would be when researchers conducted a case-control study examining whether exposure to diesel exhaust particles increases the risk of respiratory disease in underground miners. Cases included miners diagnosed with respiratory disease, while controls were miners without respiratory disease. Participants' past occupational exposures to diesel exhaust particles were evaluated to compare exposure rates between cases and controls.
A cross-sectional study (also sometimes called a "cross-sectional survey") serves as an observational tool, where researchers capture data from a cohort of participants at a singular point. This approach provides a 'snapshot'— a brief glimpse into the characteristics or outcomes prevalent within a designated population at that precise point in time. The primary aim here is not to track changes or developments over an extended period but to assess and quantify the current situation regarding specific variables or conditions. Such a methodology is instrumental in identifying patterns or correlations among various factors within the population, providing a basis for further, more detailed investigation.
A systematic review is a comprehensive approach designed to identify, evaluate, and synthesise all available evidence relevant to a specific research question. In essence, it collects all possible studies related to a given topic and design, and reviews and analyses their results.
The process involves a highly sensitive search strategy to ensure that as much pertinent information as possible is gathered. Once collected, this evidence is often critically appraised to assess its quality and relevance, ensuring that conclusions drawn are based on robust data. Systematic reviews often involve defining inclusion and exclusion criteria, which help to focus the analysis on the most relevant studies, ultimately synthesising the findings into a coherent narrative or statistical synthesis. Some systematic reviews will include a meta-analysis.
A randomised controlled trial (RCT) is a trial in which participants are randomly assigned to one of two or more groups: the experimental group or groups receive the intervention or interventions being tested; the comparison group (control group) receive usual care or no treatment or a placebo. The groups are then followed up to see if there are any differences between the results. This helps in assessing the effectiveness of the intervention.
Research that aims to gather and analyse non-numerical (descriptive) data in order to gain an understanding of individuals' social reality, including understanding their attitudes, beliefs, and motivation. This type of research typically involves in-depth interviews, focus groups, or field observations in order to collect data that is rich in detail and context. Qualitative research is often used to explore complex phenomena or to gain insight into people's experiences and perspectives on a particular topic. It is particularly useful when researchers want to understand the meaning that people attach to their experiences or when they want to uncover the underlying reasons for people's behavior. Qualitative methods include ethnography, grounded theory, discourse analysis, and interpretative phenomenological analysis.
Diagnostic accuracy studies focus on estimating the ability of the test(s) to correctly identify subjects with a predefined target condition, or the condition of interest (sensitivity) as well as to clearly identify those without the condition (specificity).
Prediction Models
Prediction model research is used to test the accurarcy of a model or test in estimating an outcome value or risk. Most models estimate the probability of the presence of a particular health condition (diagnostic) or whether a particular outcome will occur in the future (prognostic). Prediction models are used to support clinical decision making, such as whether to refer patients for further testing, monitor disease deterioration or treatment effects, or initiate treatment or lifestyle changes. Examples of well known prediction models include EuroSCORE II for cardiac surgery, the Gail model for breast cancer, the Framingham risk score for cardiovascular disease, IMPACT for traumatic brain injury, and FRAX for osteoporotic and hip fractures.
Quality improvement research is about finding out how to improve and make changes in the most effective way. It is about systematically and rigourously exploring "what works" to improve quality in healthcare and the best ways to measure and disseminate this to ensure positive change. Most quality improvement effectiveness research is conducted in hospital settings, is focused on multiple quality improvement interventions, and uses process measures as outcomes. There is a great deal of variation in the research designs used to examine quality improvement effectiveness.
A meta-analysis is a statistical technique that amalgamates data from multiple studies to yield a single estimate of the effect size. This approach enhances precision and offers a more comprehensive understanding by integrating quantitative findings. Central to a meta-analysis is the evaluation of heterogeneity, which examines variations in study outcomes to ensure that differences in populations, interventions, or methodologies do not skew results. Techniques such as meta-regression or subgroup analysis are frequently employed to explore how various factors might influence the outcomes. This method is particularly effective when aiming to quantify the effect size, odds ratio, or risk ratio, providing a clearer numerical estimate that can significantly inform clinical or policy decisions.
How Meta-analyses and Systematic Reviews Work Together
Systematic reviews and meta-analyses function together, each complementing the other to provide a more robust understanding of research evidence. A systematic review meticulously gathers and evaluates all pertinent studies, establishing a solid foundation of qualitative and quantitative data. Within this framework, if the collected data exhibit sufficient homogeneity, a meta-analysis can be performed. This statistical synthesis allows for the integration of quantitative results from individual studies, producing a unified estimate of effect size. Techniques such as meta-regression or subgroup analysis may further refine these findings, elucidating how different variables impact the overall outcome. By combining these methodologies, researchers can achieve both a comprehensive narrative synthesis and a precise quantitative measure, enhancing the reliability and applicability of their conclusions. This integrated approach ensures that the findings are not only well-rounded but also statistically robust, providing greater confidence in the evidence base.
Why Don't All Systematic Reviews Use a Meta-Analysis?
Systematic reviews do not always have meta-analyses, due to variations in the data. For a meta-analysis to be viable, the data from different studies must be sufficiently similar, or homogeneous, in terms of design, population, and interventions. When the data shows significant heterogeneity, meaning there are considerable differences among the studies, combining them could lead to skewed or misleading conclusions. Furthermore, the quality of the included studies is critical; if the studies are of low methodological quality, merging their results could obscure true effects rather than explain them.
Protocol
A plan or set of steps that defines how something will be done. Before carrying out a research study, for example, the research protocol sets out what question is to be answered and how information will be collected and analysed.
When ARRIVE refers to animal research it is referring to in vivo animal research. This is the use of non-human animals, sometimes known as model organisms, in experiments that seek to control the variables that affect the behavior or biological system under study. This approach can be contrasted with field studies in which animals are observed in their natural environments or habitats. Animal research varies on a continuum from pure research, focusing on developing fundamental knowledge of an organism, to applied research, which may focus on answering some questions of great practical importance, such as finding a cure for a disease. Source"
The ARRIVE guidelines apply to all areas of bioscience research involving living animals. That includes mammalian species as well as model organisms such as Drosophila or Caenorhabditis elegans. Each item is equally relevant to manuscripts centred around a single animal study and broader-scope manuscripts describing in vivo observations along with other types of experiments. The exact type of detail to report, however, might vary between species and experimental setup; this is acknowledged in the guidance provided for each item. Source
Bias
The over- or underestimation 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.\n\nSource
Descriptive and inferential statistics
Descriptive statistics are used to summarise the data. They generally include a measure of central tendency (e.g., mean or median) and a measure of spread (e.g., standard deviation or range). Inferential statistics are used to make generalisations about the population from which the samples are drawn.
Hypothesis tests such as ANOVA, Mann-Whitney, or t tests are examples of inferential statistics.\n\nSource
Effect size
Quantitative measure of differences between groups, or strength of relationships between variables.\n\nSource
Experimental unit
Biological entity subjected to an intervention independently of all other units, such that it is possible to assign any two experimental units to different treatment groups. Sometimes known as unit of randomisation.\n\nSource
External validity
Extent to which the results of a given study enable application or generalisation to other studies, study conditions, animal strains/species, or humans.\n\nSource
False negative
Statistically nonsignificant result obtained when the alternative hypothesis (H~1~) is true. In statistics, it is known as the type II error.\n\nSource
False positive
Statistically significant result obtained when the null hypothesis (H~0~) is true. In statistics, it is known as the type I error.\n\nSource
Independent variable
Variable that either the researcher manipulates (treatment, condition, time) or is a property of the sample (sex) or a technical feature (batch, cage, sample collection) that can potentially affect the outcome measure. Independent variables can be scientifically interesting, or nuisance variables. Also known as predictor variable.\n\nSource
Internal validity
Extent to which the results of a given study can be attributed to the effects of the experimental intervention, rather than some other, unknown factor(s) (e.g., inadequacies in the design, conduct, or analysis of the study introducing bias).\n\nSource
Nuisance variable
Variables that are not of primary interest but should be considered in the experimental design or the analysis because they may affect the outcome measure and add variability. They become confounders if, in addition, they are correlated with an independent variable of interest, as this introduces bias. Nuisance variables should be considered in the design of the experiment (to prevent them from becoming confounders) and in the analysis (to account for the variability and sometimes to reduce bias). For example, nuisance variables can be used as blocking factors or covariates.\n\nSource
Null and alternative hypotheses
The null hypothesis (H~0~) is that there is no effect, such as a difference between groups or an association between variables. The alternative hypothesis (H~1~) postulates that an effect exists.\n\nSource
Outcome measure
Any variable recorded during a study to assess the effects of a treatment or experimental intervention. Also known as dependent variable, response variable.\n\nSource
Power
For a predefined, biologically meaningful effect size, the probability that the statistical test will detect the effect if it exists (i.e., the null hypothesis is rejected correctly).\n\nSource
Sample size
Number of experimental units per group, also referred to as n.\n\nSource
