All serum samples collected from villagers and domestic animals were tested for Leishmania antibody by DAT (Biomedical Research, The Netherlands) at the Department of Parasitology, Phramongkutklao College of Medicine, following the manufacturer’s instructions. DNA extraction DAT-positive blood was performed following the manufacturer’s instructions (Qiagen, Germany) and confirmed for Leishmania infection using specific PCR conditions as previously described.