They were kept in a250ml beaker containing 150ml fish water  along with drug and observed for lethality for 24hrs. Based on the mortality, dose was increased or decreased by the factor log0.5 or 3.2.
Adult fish were obtained from the local suppliers and kept in large tanks for acclimatization. On the day of experiment, fish were taken out and they were divided into 9 treatment groups: 6 fish in each group. They were anesthetized using 0.1% 2-phenoxyethanol . Their fin was cut up to 50% by using sterilized straight razor blade.
They were again recovered using fish water. Pre and post amputated images were collected and length was measured using scale. 250ml capacity beakers were taken. Upto 150ml filled with fish water which is suspended with the drug, fish were kept in them, one fish per beaker with continuous aeration. Test drug, standard drug (Imatinib) and control were given to each group respectively. Dose for test and standard were determined from the toxicity value . On every alternate day, water was refreshed with the dose up to 15