Ex situ breeding has been identified as a key action for conserving the Endangered Owston’s civet (Chrotogale owstoni). Historically, however, the species has had poor reproductive rates under managed care. In males, it is necessary to understand testicular function to identify factors that affect reproductive success. Here, we conducted long-term faecal testosterone (Tt) monitoring, ultrasonography of the reproductive tract, and sperm assessments in five Owston’s civets held in zoos in the United Kingdom to gain insight into male fertility. Male Tt metabolite concentrations were higher in the winter and spring months, with significant cyclical increases in Tt from December to January, corresponding to the start of the in situ breeding season. No significant reproductive pathologies were identified among males, with the exception of mild irregular margins and hyperechoic foci in the prostate of one male. Semen was collected through urethral catheterization and electroejaculation from three out of four males, with repeat samples collected from one male. Urethral catheterization yielded small but highly concentrated samples, and is a simple, cost-effective, and field-friendly method of sperm collection in the species. We highlight the utility of reproductive monitoring to gain insight into male fertility.