Background: Human toxocariasis is a parasitic zoonosis caused by a parasite in the genus Toxocara and is transmitted mainly by the accidental ingestion of embryonated Toxocara canis (dog roundworm) or T. cati (cat roundworm) eggs. Several studies reported that children were the main population at risk for T. canis infection. Currently, no reports on the seroprevalence of T. canis infection in Thailand are available, and its status remains unknown among children who live in rural areas of southern Thailand. Objective: This study aimed to investigate the seroprevalence of T. canis infection and its associated risk factors among primary schoolchildren in rural areas of Nakhon Si Thammarat Province, Thailand. Methods: A total of 177 schoolchildren between 6 and 13 years of age were recruited between June and July 2019. Serum anti-T. canis IgG antibodies were detected with a commercial ELISA kit. A questionnaire administered by direct interviews was used to collect demographic and behavioral risk factor data. Results: The overall seroprevalence of T. canis infection was 58.2% (103 of 177). The univariate analysis revealed that schoolchildren who did not practice handwashing before a meal (crude odds ratio (COR) = 3.67, 95% CI 1.93-6.95, P < 0.001), did not practice hand washing after animal contact (COR = 2.89, 95% CI 1.53-5.47, P = 0.001), and drank untreated water (COR = 1.87, 95% CI 1.00-3.48, P = 0.049) had an increased risk of acquiring T. canis infection. However, after adjusting for confounders, only a lack of handwashing before a meal remained a significant risk factor (adjusted odds ratio (AOR) = 2.20, 95% CI 1.11-4.34, P = 0.023). Age, sex, owning a dog, and eating fresh vegetables were not significantly associated with T. canis infection in the current study. Conclusions: This is the first serological investigation of T. canis infection among schoolchildren in Thailand. The high rate of Toxocara seropositivity reflected high levels of exposure to T. canis among schoolchildren in rural areas of southern Thailand. The results also provide baseline data regarding modifiable risk behaviors for effective T. canis infection prevention strategies in southern Thailand, especially strengthening hand washing practices among schoolchildren.