Naturally captured Lanchester's freshwater prawns (Macrobrachium lanchesteri) and farm-bred Rana regulosa tadpoles were assessed for their capability of being the first or second intermediate hosts of Gnathostoma spinigerum. Seventy specimens from each animal group were randomized into a control group and investigated for larvae of G. spinigerum by pressing them between two pieces of glass and examination under stereomicroscope. No Gnathostoma larvae were found in the entire control group. Another 120 specimens of each animal were used in two treatment groups; 60 being exposed to the first-stage larvae, G. spinigerum, and 60 exposed to cyclops containing the third-stage larvae for 7 days. No larvae of G. spinigerum were found in the prawns of both treatment groups that resembled tadpoles exposed to the first-stage larvae. In contrast, 18.3% of tadpoles, which were exposed to cyclops containing third-stage larvae, were infected. Lanchester's freshwater prawns cannot serve as intermediate host of G. spinigerum, while R. regulosa can serve as the second intermediate host.