The present study was carried out to study the effect of different doses of hemp seed alone or in combination with dill seed against antibiotic growth promoter on performance, serum biochemicals and gut health of broiler chickens over a period of 42 days. Total 192 broiler chicks were grouped randomly into six treatments and fed with basal diet (BD) along with different levels of seeds, viz., T1 (BD), T2 (BD + 0.2% HS), T3 (BD + 0.2% HS + 0.3 DS), T4 (BD + 0.3% HS) and T5 (BD + 0.3% HS + 0.3 DS) and T6 (BD + 0.025% Bacitracin Methylene Disalicylate-BMD). The performance traits like feed intake, body weight gain and feed conversion ratio (FCR) and carcass traits like cut-up parts, giblet and abdominal fat yield remained unaffected due to dietary treatments for overall trial period; however, the average feed intake in early phase (0-3 weeks) reduced significantly (p < 0.05) in treatment birds than both controls (T1 & T6). Serum protein concentration remained unchanged, whereas significant (p < 0.05) reduction in serum lipids like triglyceride, LDL and total cholesterol concentration was noticed due to dietary inclusion of seeds. Serum enzymes like AST and ALT concentrations depleted significantly (p < 0.05) treated groups, except at higher seed doses (T5); however, alkaline phosphatase levels were unaffected. Coliform count in caecum and jejunum reduced linearly (p < 0.01) due to seed inclusion, whereas dose-dependent proliferation of lactobacilli was evident (p < 0.01) in caecum and jejunum of treated birds. No effect was observed on the villus height and crypt depth of the jejunal mucosa. To conclude, dietary supplementation of hemp and dill seed could not affect the growth performance and carcass traits; however, it positively altered the serum lipid profile of the birds and improved gut health as well, thereby enhanced overall performance of broiler chickens.