We studied the water quality of the riparian firefly sanctuary of Sungai Rembau, or Rembau River, in Negeri Sembilan, Malaysia, from January 2018 to November 2018 to determine the possible influence of the physico-chemical characteristics of the water on the firefly populations living within the sanctuary. We set up a total of five water quality sampling stations and 10 firefly sampling stations along the river. Dissolved oxygen (DO), temperature, pH and electrical conductivity (EC) were measured in situ, while chemical oxygen demand (COD), total suspended solids (TSS), biochemical oxygen demand (BOD) and ammonia-nitrogen (NH3-N) were analysed in the laboratory. Firefly samples were collected using a sweep net at both day and night for 1 min. Sungai Rembau was categorized as Class II on the Malaysian water quality index (WQI), which indicates slight pollution. Except for EC and DO, the water quality parameter values were not significantly different (p > 0.05) between the sampling stations. A total of 529 firefly individuals consisting of Pteroptyx tener (n = 525, 99.24%), P. malaccae (n = 3, 0.57%) and P. asymmetria (n = 1, 0.19%) were collected. There was significant correlation between firefly abundance and BOD (r = - 0.198, p