Abstract Gut microbiomes are crucial in determining wildlife’s health, especially in the captive population. The alternation of environment can lead to microbiome changes that could affect the host’s health. Therefore, the information regarding gut microbiomes is important in conducting effective husbandry management for captive animals especially in zoos. Thus, we conducted the gut microbiome study on captive leopard cat (Prionailurus bengalensis) using Illumina Miniseq platform. The 16S rRNA gene sequencing was performed on two samples from both male and female leopard cats. The sequencing analysis successfully unveiled the diversity of gut microbiomes within this species, identifying members from 12 phyla, 74 families, and 127 genera of bacteria. Proteobacteria and Firmicutes emerged as the dominant phyla, constituting 49% in females and 34.7% in males, respectively. Intriguingly, no significant differences in alpha diverisity indices were observed between the sexes of P. bengalensis. These findings contribute substantially to our comprehension of the gut microbiome diversity in leopard cats in Malaysia. Generally, the findings of this study provide the fundamental data for the captive management and health assessment of P. bengalensis, acting as a valuable resource for future conservation efforts. By establishing a baseline understanding of the gut microbiomes in captivity, this research supports the broader goal of preserving the biological integrity of leopard cat populations especially in Malaysia.