Mithun, a semidomesticated and autochthonous bovine species of northeastern India, is reared under a free-range forest system.The 20 th livestock census shows an increase of 29.5% (3-3.9 lakhs) mithun over the previous census (20 th Livestock census, DAHD, Government of India, 2019). However, in recent times, denuding forest cover, commercialization of cropping systems, cross-breeding and sacrifice of potential elite mithun bulls have significantly decreased its habitat and the elite mithun population. The diminished population of superior breedable mithun bulls has increased the inbreeding within the population resulting in various reproductive problems including infertility. Therefore, elite bull selection, semen collection and artificial breeding programme need to be taken to overcome the inbreeding depression and to conserve and propagate this precious mountain bovine species.To have germplasm conservation, there is a need for an effective semen collection method from free-range mithun bulls. The rectal massage (Bhattacharyya et al., 2005) and artificial vagina (AV) (Bhattacharyya et al., 2009) methods have been used in collecting ejaculates from the farm mithun. Semen collected from the rectal massage method is poor in quality and quantity, resulted in