DEVELOPMENT OF LAPAROSCOPIC OVUM PICK-UP (LOPU)
We first published the development of the LOPU technique for oocyte collection in sheep in 1994 (Baldassarre et al.; Anim. Reprod. Sci. (35), 145-150). Since then we have conducted the procedure in thousands of sheep and goats with an average of 10 and 14 oocytes/donor, respectively. We have optimized the technique over the last 20 years and it has become the standard of excellence for in vitro embryo production, production of zygotes for microinjection (transgenic generation) and the production of recipient cytoplasts for nuclear transfer (cloning). In addition, the technique has allowed us to produce offspring from prepubertal animals (1-3 months of age) in sheep, goats, cattle and buffalo. Outstanding milestones: First IVF lamb born in Argentina (CIRPC, 1995), First IVF goat kids born in Canada (Nexia, 1999), First IVF lamb in Brazil (In Vitro Brazil, 2006), first IVF buffalo in Canada (2018). More recently we have expanded the use of LOPU to Marmosets and Swine, specifically for the generation of Animal Models for Biomedical applications.
PREPUBERTAL REPRODUCTION IN SHEEP, GOATS, CATTLE, and growing...
Through the application of LOPU technology for the collection of oocytes followed by in vitro embryo production and embryo transfer into adult recipients, we have produced large numbers of offspring from pre-pubertal sheep and goats (1-3 months of age). Some of these animals were first of their kind in Argentina and Canada. This development is important because the progeny is born before the mother reaches sexual maturity and can be bred for the first time. In addition to accelerating genetic progress by means of shortening the generation interval, pre-pubertal females are fantastic oocyte donors with average productivities of 40 oocytes/donor. In combination with the availability of genetic markers that allow predicting individual productivity from birth, this technology is expected to play an important role in the future of livestock improvement. We have also successfully produced offspring from pre-pubertal female calves using the same technology developed for sheep and goats. Outstanding milestones: We produced the first calves from embryos produced from prepubertal calves in Latin America (CIRPC, 1995), and the world's first goat progeny from oocytes collected from live prepubertal goats (Nexia 1999). More recently we incorporated whitetail deer and buffalos to the list of species propagated at prepubertal ages using LOPU.
PRODUCTION OF CLONED SHEEP & GOATS
In Canada, we produced >250 cloned goats (mostly transgenic Saanens) of which >80% lived to become adults. In Argentina we produced additional cloned goats (Boers) as well as cloned sheep of three different breeds (Dorper, Santa Inés and Texel). Outstanding milestones: The above-referred efforts included the first cloned goats in Canada (Nexia, 1999), the first cloned goats in Latin America (Germinal Biotech, 2010), and the first live cloned sheep in Brazil (Fortaleza, 2014).
TRANSGENIC GOAT PRODUCTION
From 1998 to 2011, I was in charge of the development of all animal-related aspects of what was possibly the world's biggest herd of transgenic goats (>800 transgenic goats at peak). This included the use of ART for founder generation and propagation of transgenic goats (MOET, IVEP, JIVEP, SCNT, AI), as well as all elements of Farm Operations under GMP-like practices (Animal Husbandry, Animal Health, Quality Assurance, etc.). All goats had incorporated a DNA cassette into their genome instructing the expression of a recombinant protein of pharmaceutical interest in the milk. In the most successful program, goats expressed human butyrylcholinesterase (rBChE) in the milk, a potent antidote against organophosphate poisoning (e.g. nerve agents). Outstanding milestone: In 2006, the US-Department of Defense awarded us a contract for $213 million dollars for the development of 50,000 doses of rBChE.