Background Leptospirosis is a bacterial infection caused by spirochetes of the genusLeptospira. It affects both humans and animals, posing a significant public health threat worldwide. In Sri Lanka, leptospirosis causes annual outbreaks, particularly during the monsoon seasons. The disease is often difficult to diagnose and can have a high mortality rate. Bats are potential reservoirs forLeptospirain other countries. This study investigated the presence and diversity ofLeptospirain bats inhabiting the Wavulgalge, a natural cave in Sri Lanka. Methodology/Principal Findings Using PCR and Sanger sequencing techniques,Leptospira borgpeterseniiandLeptospira kirschneriwere identified inMiniopterus fuliginosusandRousettus leschenaultiibats. Out of 148 bat urine samples, 16 tested positive via RT-qPCR, withM. fuliginosussamples harboringL. borgpeterseniiand oneR. leschenaultiisample harboringL. kirschnerirevealed by Sanger sequencing method. Conclusions/Significance These findings enhance the understanding of the zoonotic role of Sri Lankan bats inLeptospiratransmission.