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Volume 8, Issue 4 (Suppl) J Vet Sci Technol, an open access journal
ISSN: 2157-7579 Veterinary 2017
September 04-05, 2017
September 04-05, 2017 | Paris, France
7th International Veterinary Congress
Production of pashmina (cashmere) goat through handmade cloning technique using continuous
culture system
Riaz Shah, Abrar Malik, Maajid Bhat, Firdous Khan, Syed Hilal, Mujeeb Fazili, Mudasir Andrabi, Nadeem Shabir and Nazir Ganai
SKUAST Kashmir, India
Statement of the Problem: Pashmina (cashmere) goat is among the few species of livestock which can survive in their inhabitant
harsh cold arid climate. This goat produces world’s finest fibre. Due to high risk of genetic loss via inbreeding, reproductive
technologies need to be implemented. Cloning is the fastest way to multiply the limited superior germplasm. Handmade
cloning (HMC) is an alternative method of cloning which eliminates the use of costly sophisticated micromanipulator tools
demanding greater degree of skill, in comparison to SCNT. The present study was designed to optimize in vitro continuous
culture system for development of zona free handmade cloned pashmina goat blastocysts and they’re in-vivo development after
laparoscope aided intra uterine transfer into synchronized recipients.
Methodology &Theoretical Orientation: Skin derived fibroblasts at 5th to 9th passage was used as nucleus donor cells for HMC
experiments. Cumulus–oocyte complexes (COCs) were in vitro matured and stripped of their cumulus investment and zona
pellucida. Protrusion cone-guided bisection was performed for enucleation. Electro-fusion was carried out to generate triplets
(two demicytoplasts and a donor cell). The reconstructed zygotes were then activated and cultured in different experimental
groups wherein we compared different culture media and culture systems. The blastocysts were transferred into synchronized
recipient goats by laparoscope aided transfer technique. Pregnancies were diagnosed through USG after 45 days of transfer.
Identification and confirmation of the clone born was performed via microsatellite marker analysis.
Findings: The cleavage and blastocyst rates were determined at day 7 of embryo culture. G1. G2 medium gave the best cleavage
percentage (86.84±2.26) while as RVCL (commercial medium from CookTM, Australia) gave the best blastocyst percentage
(15.01±4.58). WID (well in drop) culture systemwas found to be most efficient with highest cleavage and blastocyst percentages
i.e., 84.34±4.15 and 21.65±1.69 respectively. On day 45 post embryo transfer into 19 receipients, 3 pregnancies were detected
out of which only one carried to term.
Conclusion & Significance: Using cost effective HMC technique, we successfully report the live birth of first handmade cloned
cashmere goat. The birth weight of the cloned kid was 2.4 kg, like female kids from naturally bred Pashmina goats during the
same period. No significant differences in growth rate between cloned goat and naturally bred goats (1.2 kg/month) were
observed till it reached sexual maturity. Also, estrous cycle of the cloned goat was observed to be normal and bred normally.
In our study, the embryos were cultured in a serum free media which could explain the normal birth weight of the cloned kid.
Microsatellite analysis confirmed that the cloned kid was genetically identical to the fibroblast cell donor dam. In conclusion,
this study elucidated the production of hand-made cloned blastocysts using a continuous culture system and birth of healthy
cloned kid from Pashmina goat
Biography
Riaz Shah has completed his Graduation in Veterinary Sciences and Master’s/PhD degree in Animal Biotechnology with specialization in Livestock Reproductive
Biotechnology and Post-doctoral experience from AgResearch Ltd., Ruakura Research Centre, Hamilton, Newzealand in area of goat cloning. His current areas
of research are the Application of Advanced Reproductive Technologies like IVF, SCNT and Stem Cell Production for augmenting production of Livestock. The
recent successes in cloning of livestock (Buffalo and Pashmina goat) first time in India through Handmade Cloning technique during his research endeavors at
National Dairy Research Institute, Haryana and SKUAST-K, Kashmir, India, has opened opportunities to achieve excellence in the application of such techniques
for transgenic animal production and stem cell research. He is currently working as a Professor and Head, Division of Animal Biotechnology at Faculty of Veterinary
Sciences in SK Agricultural University, Srinagar, J&K, India.
Riaz Shah et al., J Vet Sci Technol 2017, 8:4(Suppl)
DOI: 10.4172/2157-7579-C1-024