16. Animal care and monitoring
A safe and effective analgesic plan is critical to relieve pain, suffering and distress. Untreated pain can affect the animals’ biology and add variability to the experiment; however specific pain management procedures can also introduce variability, affecting experimental data [1,2]. Under-reporting of welfare management procedures contributes to the perpetuation of non-compliant methodologies and insufficient or inappropriate use of analgesia  or other welfare measures. A thorough description of the procedures used to alleviate pain, suffering and distress provides practical information for researchers to replicate the method.
Clearly describe pain management strategies, including:
- specific analgesic
- administration method (e.g. formulation, route, dose, concentration, volume, frequency, timing, and equipment used)
- rationale for the choice (e.g. animal model, disease/pathology, procedure, mechanism of action, pharmacokinetics, personnel safety)
- protocol modifications to reduce pain, suffering and distress (e.g. changes to the anaesthetic protocol, increased frequency of monitoring, procedural modifications, habituation, etc.)
If analgesics or other welfare measures, reasonably expected for the procedure performed, are not performed for experimental reasons, report the scientific justification .
- Jirkof P (2017). Side effects of pain and analgesia in animal experimentation. Lab Anim (NY). doi: 10.1038/laban.1216
- Carbone L and Austin J (2016). Pain and Laboratory Animals: Publication Practices for Better Data Reproducibility and Better Animal Welfare. PLoS One. doi: 10.1371/journal.pone.0155001
- Gaspani L, Bianchi M, Limiroli E, Panerai AE and Sacerdote P (2002). The analgesic drug tramadol prevents the effect of surgery on natural killer cell activity and metastatic colonization in rats. Journal of neuroimmunology. doi: 10.1016/s0165-5728(02)00165-0
“If piglets developed diarrhea, they were placed on an electrolyte solution and provided supplemental water, and if the diarrhea did not resolve within 48 h, piglets received a single dose of ceftiofur (5.0 mg ceftiofur equivalent/kg of body weight i.m [Excede, Zoetis, Florham Park, NJ]). If fluid loss continued after treatment, piglets then received a single dose of sulfamethoxazole and trimethoprim oral suspension (50 mg/8 mg per mL, Hi-Tech Pharmacal, Amityville, NY) for 3 consecutive days.” 
“One hour before surgery, we administered analgesia to the mice by offering them nut paste (Nutella; Ferrero, Pino Torinese, Italy) containing 1 mg per kg body weight buprenorphine (Temgesic; Schering-Plough Europe, Brussels, Belgium) for voluntary ingestion, as described previously. The mice had been habituated to pure nut paste for 2 d prior to surgery.” 
“If a GCPS score equal or greater than 6 (out of 24) was assigned postoperatively, additional analgesia was provided with methadone 0.1 mg kg−1 IM (or IV if required)…and pain reassessed 30 minutes later. The number of methadone doses was recorded.” 
- Getty CM and Dilger RN (2015). Moderate Perinatal Choline Deficiency Elicits Altered Physiology and Metabolomic Profiles in the Piglet. PLoS One. doi: 10.1371/journal.pone.0133500
- Teilmann AC, Falkenberg MK, Hau J and Abelson KS (2014). Comparison of silicone and polyurethane catheters for the catheterization of small vessels in mice. Lab Anim (NY). doi: 10.1038/laban.570
- Bustamante R, Daza MA, Canfrán S, García P, Suárez M, Trobo I and Gómez de Segura IA (2018). Comparison of the postoperative analgesic effects of cimicoxib, buprenorphine and their combination in healthy dogs undergoing ovariohysterectomy. Veterinary Anaesthesia and Analgesia. doi: 10.1016/j.vaa.2018.01.003
Reporting adverse events allows other researchers to plan appropriate welfare assessments and minimise the risk of these events occurring in their own studies. If the experiment is testing the efficacy of a treatment, the occurrence of adverse events may alter the balance between treatment benefit and risk .
Report any adverse events that had a negative impact on the welfare of the animals in the study (e.g. cardiovascular and respiratory depression, CNS disturbance, hypothermia, reduction of food intake). Indicate whether they were expected or unexpected. If adverse events were not observed, or not recorded during the study, explicitly state this.
- Muhlhausler BS, Bloomfield FH and Gillman MW (2013). Whole Animal Experiments Should Be More Like Human Randomized Controlled Trials. PLoS Biol. doi: 10.1371/journal.pbio.1001481
“Murine lymph node tumors arose in 11 of 12 mice that received N2-transduced human cells. The neo gene could be detected in murine cells as well as in human cells. Significant lymphoproliferation could be seen only in the murine pre-T cells. It took 5 months for murine leukemia to arise; the affected mice displayed symptoms of extreme sickness rapidly, with 5 of the 12 mice becoming moribund on exactly the same day (Figure…), and 6 others becoming moribund within a 1-month period…Of the 12 mice that had received N2-transduced human cells, 11 had to be killed because they developed visibly enlarged lymph nodes and spleen, hunching, and decrease in body weight, as shown in Figure…The 12th mouse was observed carefully for 14 months; it did not show any signs of leukemia or other adverse events, and had no abnormal tissues when it was autopsied…The mice were observed at least once daily for signs of illness, which were defined as any one or more of the following: weight loss, hunching, lethargy, rapid breathing, skin discoloration or irregularities, bloating, hemi-paresis, visibly enlarged lymph nodes, and visible solid tumors under the skin. 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.” 
“Although procedures were based on those reported in the literature, dogs under Protocol 1 displayed high levels of stress and many experienced vomiting. This led us to significantly alter procedures in order to optimize the protocol for the purposes of our own fasting and postprandial metabolic studies.” 
- Bauer G, Dao MA, Case SS, Meyerrose T, Wirthlin L, Zhou P, Wang X, Herrbrich P, Arevalo J, Csik S, Skelton DC, Walker J, Pepper K, Kohn DB and Nolta JA (2008). In vivo biosafety model to assess the risk of adverse events from retroviral and lentiviral vectors. Mol Ther. doi: 10.1038/mt.2008.93
- Bellanger S, Benrezzak O, Battista MC, Naimi F, Labbe SM, Frisch F, Normand-Lauziere F, Gallo-Payet N, Carpentier AC and Baillargeon JP (2015). Experimental dog model for assessment of fasting and postprandial fatty acid metabolism: pitfalls and feasibility. Lab Anim. doi: 10.1177/0023677214566021
Humane endpoints are predetermined morphological, physiological and/or behavioural signs that define the circumstances under which an animal will be removed from an experimental study. The use of humane endpoints can help minimise harm while allowing the scientific objectives to be achieved . Report the humane endpoints that were established for the specific study, species and strain. Include clear criteria of the clinical signs monitored , and clinical signs that led to euthanasia or other defined actions. Include details such as general welfare indicators (e.g. weight loss, reduced food intake, abnormal posture) and procedure-specific welfare indicators (e.g. tumour size in cancer studies , sensory motor deficits in stroke studies ).
Report the timing and frequency of monitoring, taking into consideration the normal circadian rhythm of the animal and timing of scientific procedures, as well as any increase in the frequency of monitoring (e.g. post-surgery recovery, critical times during disease studies, or following the observation of an adverse event). Publishing score sheets of the clinical signs that were monitored  can help guide other researchers to develop clinically relevant welfare assessments, particularly for studies reporting novel procedures.
This information should be reported even if no animal reached any of the humane endpoints. If no humane endpoints were established for the study, explicitly state this.
- Hendriksen C, Morton D and Cussler K (2011). Use of humane endpoints to minimise suffering. From: The COST manual of animal care and use. CRC Press, Florida, USA. https://www.cost.eu/publications/the-cost-manual-of-laboratory-animal-care-and-use-refinement-reduction-and-research/
- Hawkins P, Morton DB, Burman O, Dennison N, Honess P, Jennings M, Lane S, Middleton V, Roughan JV, Wells S and Westwood K (2011). 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. Laboratory Animals. doi: 10.1258/la.2010.010031
- Workman P, Aboagye EO, Balkwill F, Balmain A, Bruder G, Chaplin DJ, Double JA, Everitt J, Farningham DAH, Glennie MJ, Kelland LR, Robinson V, Stratford IJ, Tozer GM, Watson S, Wedge SR, Eccles SA and An ad hoc committee of the National Cancer Research I (2010). Guidelines for the welfare and use of animals in cancer research. British Journal Of Cancer. doi: 10.1038/sj.bjc.6605642
- Percie du Sert N, Alfieri A, Allan SM, Carswell HV, Deuchar GA, Farr TD, Flecknell P, Gallagher L, Gibson CL, Haley MJ, Macleod MR, McColl BW, McCabe C, Morancho A, Moon LD, O'Neill MJ, Perez de Puig I, Planas A, Ragan CI, Rosell A, Roy LA, Ryder KO, Simats A, Sena ES, Sutherland BA, Tricklebank MD, Trueman RC, Whitfield L, Wong R and Macrae IM (2017). The IMPROVE Guidelines (Ischaemia Models: Procedural Refinements Of in Vivo Experiments). J Cereb Blood Flow Metab. doi: 10.1177/0271678X17709185
- Morton DB (2000). A systematic approach for establishing humane endpoints. ILAR Journal. doi: 10.1093/ilar.41.2.80
“Both the research team and the veterinary staff monitored animals twice daily. Health was monitored by weight (twice weekly), food and water intake, and general assessment of animal activity, panting, and fur condition…The maximum size the tumors allowed to grow in the mice before euthanasia was 2000 mm3.” 
- Muscella A, Vetrugno C, Cossa LG, Antonaci G, De Nuccio F, De Pascali SA, Fanizzi FP and Marsigliante S (2016). In Vitro and In Vivo Antitumor Activity of [Pt(O,O'-acac)(gamma-acac)(DMS)] in Malignant Pleural Mesothelioma. PLoS One. doi: 10.1371/journal.pone.0165154