Introduction: Leptospirosis is a widespread zoonotic disease, which has a medical and veterinary importance, and also a commonly re-emerging infectious disease. The main causes of transmission are known; however, the respective prominence of each of the components and the respective environmental risk factors are obscure. Methodology: Present study was conducted in the different locations (urban/rural/tribal) of the union territory of Dadra and Nagar Haveli (UT of Dadra and Nagar Haveli). Periodical sample collection approach was used to collect the samples from May 2016 to April 2017, to determine the persistence of leptospiral contamination of the environmental sources. Results: The PCR detection and isolation of Leptospira revealed that pathogenic leptospires were present in water, soil and animal urine. The highest positivity was reported from the water of household drainage in urban areas and in the water from rice field of rural areas. Conclusions: The data obtained from the present study may help and guiding in developing preventive measures for leptospirosis in the UT of Dadra and Nagar Haveli.