Liver fluke (Opisthorchis viverrini, O.v.) infection, along with its associated cholangiocarcinoma, is a major public health problem in Southeast Asia. Despite the vast amount of epidemiological research, human O.v. prevalence remains high and varies greatly across the region. This paper examines the landscape determinants that influence O.v. transmission in relation to the three hosts of its life cycle and identifies areas that require further research so as to advance the understanding of the spatial variation in disease risk. A critical agent functionally connects all sequential life cycle stages of O.v. is water. Seasonality and water quality appear to affect the habitats and population dynamics of the two intermediate hosts, Bithynia snails and cyprinid fish. Land use practice through the construction of irrigation ditches increases the connections between the hosts, thereby functionally facilitating the disease transmission. Multi-season sampling data of host infections and habitat characteristics are needed for integration with analyses of landscape connectivity and human behavior to allow better understanding of the interactions among the landscape determinants on the spatial-temporal dynamics of disease transmission.