Leptospirosis is a neglected tropical disease that affects mostly rural people in Thailand. This study aims to identify the factors explaining leptospirosis epidemiology, to describe the health policy implemented for leptospirosis prevention and control and its implementation in the rural province of Mahasarakham (Thailand), and to analyse the representations, practices, and attitudes of patients, people, health volunteers and health officers toward the disease and its prevention.The analysis of health policies showed that Thailand has progressively developed a national prevention and control policy framework and transferred its implementation to local administrative levels. However, leptospirosis prevention and control still need enhanced collaboration between public health, livestock, and agriculture departments with an important local community involvement under the One Health approach.The analysis of the spatiotemporal distribution of human leptospirosis cases showed that the factors associated with leptospirosis infection are complex and multifactorial, including human population density, livestock density, rainfall, flood cover and physical geography, i.e. average slope. The results stressed the importance of livestock that may contribute to leptospirosis transmission to humans. Targeting areas prone at risk, i.e. with high livestock or in flooded areas, should be complemented by improving communication to people at risk, i.e. farmers.The cross sectional study conducted to assess knowledge, attitudes, and practices showed that patients, neighbors, village health volunteers and community leaders did not have a clear comprehension of the disease and its transmission to really improve the prevention of infections. People need to have access to a complex and up-to-date scientific knowledge in order to improve their awareness about leptospirosis.Finally, taking results of above issues allowed “Guidelines for developing cooperation between departments for prevention and control leptospirosis of Mahasarakham Province, Thailand” to be produced.