For the definitive test, the concentration used range from 3.125% to 5.625% v/v of leachate. Similarly,
for each concentration namely 5.625%, 5.00%, 4.375%, 3.750% and 3.125%, 10 fishes were introduced.
A control was set-up without the addition of leachate. The mortality and change in the fish behavior were
recorded for every 12 hours. The LC50 of A. testudineus was calculated using EPA Probit software.