Bluetongue virus (BTV) remains as an economically major concern in the world. Seroprevalence and potential risk factors of BTV were assessed in a cross-sectional study at both the herd and animal levels in Iran. A total of 73 Epidemiologic Units (E.Unit), defined as a herd, flock or village including animals with equal chance of exposure to infectious agents, were randomly selected. Serum samples from all animals (n = 34,575) within the E.Units were collected and tested for BTV sero-group antibodies by using commercially competitive ELISA test. Using cluster analysis, 90.41 % (95 %, CI: 80.85 %–95.47 %) of the E.Units and 56.13 % (95 % CI: 55.61 %–56.66 %) of the tested animals were detected seropositive against BTV. A seroprevalence rate of 57.59 % (95 % CI: 48.01 %–66.63 %), 65.65 % (95 % CI: 59.10 %–73.74 %) and 27.63 % (95 % CI: 14.40 %–46.43 %) was estimated for sheep, goats and cattle, respectively.At E.Unit (herd) level, density was identified as a great risk factor for the infection (r2 = 0.891; P = 0.000), and particularly density of cattle significantly correlated with BTV infection within the E.Units (r2 = 0.247; P = 0.019). Using multilevel logistic regression, adjusted odds ratios (ORs) were estimated at individual level. A significantly less risk of BTV infection was evaluated for cattle than for sheep (OR = 0.42, 95 % CI: 0.38−0.47, P < 0.001), while no significant difference was observed between sheep and goat (OR = 1.03, 95 % CI: 0.97–1.10, P = 0.345). Animals over 2 years and between 6 months and 2 years expressed 2.22 (OR = 2.22, 95 % CI: 1.96−2.52, P < 0.001) and 2.18 (OR = 2.18, 95 % CI: 1.92−2.49, P < 0.001) times higher chance for the infection than animals under 6 months. Males were at significantly less risk of the infection than females (OR=0.68, 95 % CI: 0.63−0.74, P < 0.001). Animals kept in industrial farming systems displayed 0.46 (OR=0.46, 95 % CI: 032−0.66, P < 0.001) times less chance than animals kept in traditional farming system for BTV, while animals lived in semi-industrial farming system were found to be at 2.97 (OR=2.97, 95 % CI: 2.41−3.66, P < 0.001) times higher chance for BTV than animals lived in traditional farming system. Furthermore, seropositive animals exhibited a high amount of antibodies against BTV (s) suggesting that viral exposure may have frequently occurred during their lifetimes. This large - scale study yielded information on epidemiology of BTV in Iran that is prerequisite for further research, and also for evaluation of any cost-benefit control measure to be established in an enzootic zone of the virus.