1. Anees C *, and Majumdar A C Reproductive Physiology and ETT Laboratory Division of Physiology and Climatology Indian Veterinary Research Institute, Bareilly, Uttar Pradesh, India, 243122 Corresponding Author: draneescherkunnath@gmail.com

2. Population Explosion Global warming

3. Global consumption of meat- 76% Increase Global consumption of dairy- 65% increase (Rob Bailey Energy, Environment and Resources - Dec 2014) Shall we multiply the number of Animals ??? 2050: Projections (2005-07 baseline)

5. Methane CO2 Nitrous oxide

8. Hostile Climate for Production (Hot and Humid – especially in Kerala, Gujarat, Orissa and West Bengal) Few well adapted animals ( Both high producing as well as disease resistant ) Propagation of these well adapted animals by Totipotent Stem cell cloning / Culture - Concept paper (Majumdar 2005)

9. Oocyte and embryo like structures including blastocyst like structure developed from pluripotent stem cell (Majumdar et al 2002) Mouse embryonic stem cells formed oocytes, embryo and blastocyst like structure (Hubner et al 2003, Science)

10. 1.To develop embryo like structures from Totipotent stem cells (Goat Model) 2.To study the further development of embryo like structure if any, in recipient

12. In vitro fertilization (As per standard procedure in the laboratory Anees 2007) Isolation of totipotent stem cells (Standard protocol, Swain et al 2002) Culturing of these cells to develop embryo like structures

13. ELS has the potential to become an animal Embryo like structure Blastomeres In vitro Embryos The Concept of Superior Quality Animal multiplication Superior Quality Animal

14.  Selection of good quality ELS from culture  Surgical Transfer of these ‘embryos’ to synchronous recipient  Evaluation of development

16. Immature oocytes 8-16 cell embryos Mature oocytes

18. Embryo Like Structures (ELS)

19. Blastocyst like structures

20. Cells hatching out of ELS

22. Hubner et al 2003

23. Relative expression of development related genes in Embryo like structures (24 nos) in comparison to 24 number of IVF derived 8 cell stage embryos and 3 number of IVF derived 8 cell stage embryos.

24. Characterized using marker gene NKx 2.5 ANEES cardiomyocites KSC.avi

25. Two out of the three recipients returned to heat 21days post estrus One recipient returned to next cycle 28 days post estrus Further investigations are on

26.  Embryo Like Structures were developed from totipotent stem cell culture  They had similar but lower gene expression compared to IVF Embryos  Further investigation is required to improve the cell size and to produce live animal from these cells  Generation of ELS and its development can address the environmental challenges