Evaluation of ex vivo culture condition for both spermic Fasciola gigantica (sFg) and aspermic (parthenogenic) Fasciola (asF). (A) Survival of the sFg in different commercially available media. Adult sFg (2 parasites in 2 mL) were incubated in different media (M199, DMEM or RPMI 1640) containing 200 U/mL penicillin, and 200 mg/mL streptomycin in a 12-well flat bottom cell culture plate. Parasites were scored in a blinded manner, as mentioned in the Materials and Methods section. (B) Survival of asF in the commercially available media. Adult asF (2 parasites in 2 mL) were incubated following the same procedures as mentioned above and scored. (C) Adult sFg (2 parasites in 2 mL) were incubated in DMEM supplemented with bovine serum (BS), bovine bile (BB) at different concentrations (5–20%) along with 200 U/mL penicillin, and 200 mg/mL streptomycin in a 12-well flat bottom cell culture plate in the same condition, and were scored. (D) Adult asF (2 parasites in 2 mL) were incubated in DMEM supplemented with BS, BB at different concentrations following the identical procedures as mentioned above and scored. Each experiment was conducted in triplicate and data are presented as mean ± SE. *p < 0.05; **p < 0.01.