To investigate the epidemiology of tobacco use and nicotine dependence in a sample of truck drivers in Brazil. Between 2015 and 2016, a cross-sectional study was conducted on 624 truck drivers who operate on the BR-050 highway in Brazil. Participants were interviewed about sociodemographic data, occupational characteristics, mental health, behavioral data, and tobacco use. Then, the Fagerstrom test for nicotine dependence (FTND) was used to verify nicotine dependence in smoking truck drivers. Logistic regression and linear regression were also used to verify factors associated with tobacco use in the previous 30 days and nicotine dependence scores, respectively. The prevalence of tobacco use among truck drivers was 21.1% (n = 132;95%CI: 18.1-24.5). Of the total number of smokers who responded to the FTND (n = 118; 89.4%), most had high/very high nicotinic dependence (68.6%; 95%CI: 59.8-76.3). Tobacco use was associated with absence of religion (adjusted odds ratio [AOR]: 2.60; 95%CI: 1.35-5.01), employment relationship of the contract (AOR = 1.98; 95%CI: 1.26-3.13); > 12 hours daily working time (AOR = 1.80; 95%CI: 1.09-2.98) and alcohol use in the previous 30 days (AOR = 2.92; 95%CI: 1.86-4.57). Irregular physical activity was associated with higher scores of nicotine dependence (β = 1.87; 95%CI: 0.55-3.19). The results showed a high prevalence of tobacco use and high/very high nicotine dependence among the truck drivers.