Helminth infection is highly prevalent in indigenous chickens reared in semi-scavenging/ scavenging systems in Bangladesh. Here, we estimated the prevalence of gizzard worm infection in indigenous chickens, the detection of the worm-induced pathologies, the development of ex vivo cultural protocol, and anthelmintic efficacy. We randomly collected and examined 390 chickens and isolated worms from the gizzard and proventriculus. The isolated worms were identified as Cheilospirura hamulosa Diesing, 1861. The overall prevalence of C. hamulosa was 33.1% (129 out of 390). Prevalence of the worm was almost similar in both sexes but significantly (p <0.05) higher in adult chickens (44.3%) and in the summer season (47.1%). In heavy infections, C. hamulosa destroyed the muscular layer of the gizzard. The presence of brown necrotic tissues and curd-like caseous materials was detected in the affected gizzards. In severe cases, the horny lining of the gizzard was inflamed, necrotized and marked by multiple holes and brick-red colored spots. Liquefied, fetid materials oozed out from the muscular layer in extensive cases. Histopathological examination showed marked infiltrations of eosinophils. In serum-supplemented M199 and DMEM, adult C. hamulosa survived well and reproduced. Levamisole (LEV) and ivermectin (IVM) efficiently killed the worm. However, albendazole (ABZ), mebendazole (MBZ) and piperazine (PPZ) did not kill the worms. Our results suggest that C. hamulosa is highly prevalent in semi-scavenging chickens in Bangladesh. LEV and IVM can be used to treat and control the infection in chickens.