BACKGROUND: Reported monthly scrub typhus (ST) cases in Thailand has an increase in the number of cases during 2009-2014. Humidity is a crucial climatic factor for the survival of chiggers, which is the disease vectors. The present study was to determine the role of humidity in ST occurrence in Thailand and its delayed effect. METHODS: We obtained the climate data from the Department of Meteorology, the disease data from Ministry of Public Health. Negative binomial regression combined with a distributed lag non-linear model (NB-DLNM) was employed to determine the non-linear effects of different types of humidity on the disease. This model controlled overdispersion and confounder, including seasonality, minimum temperature, and cumulative total rainwater. RESULTS: The occurrence of the disease in the 6-year period showed the number of cases gradually increased summer season (Mid-February - Mid-May) and then reached a plateau during the rainy season (Mid-May - Mid-October) and then steep fall after the cold season (Mid-October - Mid-February). The high level (at 70%) of minimum relative humidity (RHmin) was associated with a 33% (RR 1.33, 95% CI 1.13-1.57) significant increase in the number of the disease; a high level (at 14 g/m3) of minimum absolute humidity (AHmin) was associated with a 30% (RR 1.30, 95% CI 1.14-1.48); a high level (at 1.4 g/kg) of minimum specific humidity (SHmin) was associated with a 28% (RR 1.28, 95% CI 1.04-1.57). The significant effects of these types of humidity occurred within the past month. CONCLUSION: Humidity played a significant role in enhancing ST cases in Thailand, particularly at a high level and usually occurred within the past month. NB-DLNM had good controlled for the overdispersion and provided the precise estimated relative risk of non-linear associations. Results from this study contributed the evidence to support the Ministry of Public Health on warning system which might be useful for public health intervention and preparation in Thailand.