Birth of India's first cloned cattle and analysis of its health and reproduction status: A case study.

Somatic cell nuclear transfer (SCNT) allows the multiplication of elite livestock and conservation of endangered species. However, restrictions on cow slaughter limit the access to oocytes for SCNT applications in Indian cattle breeds. To overcome this limitation, we utilized transvaginal ovum pick-up (OPU) method to collect oocytes, which were then used for the production of cloned embryos via the handmade cloning (HMC) technique. A total of 98 Sahiwal oocytes were collected, leading to the successful reconstruction of 24 SCNT Gir embryos. Out of these, five developed into blastocysts, which were transferred into five recipient cows. Two pregnancies were confirmed, but one was lost due to hydro-allantois condition. The other pregnancy continued to term, and a healthy Gir female calf weighing 32 kg was born. Microsatellite DNA analysis confirmed the genetic identity of the cloned calf to its donor. Postnatally, the calf was monitored for serum cytokine parameters, telomere length, and reproductive potentials. Cytokine profiling revealed variations between the cloned calf and naturally conceived counterparts; however, the born cloned calf did not exhibit any pathological conditions and has high telomere length compared to age-matched counterparts, and surviving well. Furthermore, to assess cloned cow utility in reproductive biotechnologies, we produced blastocyst stage embryos (35%) through OPU-IVF method and established one pregnancy from five transfers (20%). In conclusion, this study reports the first successful SCNT of Indian cattle breed and demonstrates the feasibility of the cloned cow for the production of OPU-IVF embryos.