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Article History

Received: 20 December 2022

Accepted: 2 March 2023

First Online: 17 March 2023

Declarations

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: The authors have no relevant financial or non-financial interests to disclose. No entity or person stands to benefit financially from the conduct of this research.

: This study was granted Ethical Approval by the Mona Campus Research Ethics Committee. Approval number AN10, 20/21. Rats were housed in sterilized plastic cages with absorbent bedding material. The metal covers used facilitated the supply of food and water ad libitum. The food, water and bedding were checked daily, and the bedding changed every 48 h. The rats were be placed in clean holding cages and the feeding cages were cleaned every 48 h. The vital statistics like temperature and weight chart were maintained. The rats were kept in normal room temperature environment which was well aerated with normal lighting present. At the end of the study, the rats were euthanised. The rats that were selected for the study weighed between 200 and 300 g. As such they were anaesthetized before euthanasia using 30 mg/kg of sodium phenobarbital. The sodium phenobarbital was administered intraperitoneally after which they were administered 120 mg/kg of phenobarbital for euthanasia. Rats were then retained in 10% formaldehyde for further studies. The ethical issue of concern for this study was to reduce the risk of unnecessary animal suffering. The research in no way abused the animals. No unnecessary procedure was undertaken. Only procedures necessary to accomplish the aim of the study were undertaken. All procedures were optimised to cause minimal pain to the animals. Other options which could have sufficiently replace the need for animal study were considered and explored such as the possibility and effectiveness of using software to simulate the probable metabolic interactions. The three R’s, Reduction, Replacement and Refinement were always considered during the study. Reduction in that during the study we re-evaluated the method based on the progress of the study to determine if less animals than originally indicated could be used to complete the study. Replacement in that during the study we explored the possibility of replacing the use of animals with non-sentient alternatives available to complete the study. Refinement in that we constantly explored ways to reduce any discomfort to the animals during the study.