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European cloning company Cryozootech announced Sept. 18 the birth of the clone of Calvaro 5, Swiss rider Villi Melliger's top-ranked jumper who died in 2003.
It took five years to clone this horse successfully. After a few failed attempts for a full-term pregnancy, a healthy colt was born in early August of this year.

Calvaro 5 won silver medals at the 1996 Atlanta and 2000 Sydney Olympic Games in show jumping, was ranked No. 1 in the world in 1999, and 2nd in 1996 and 1998. Calvaro left the jumping arena in 2002 and was retired to Melliger's stud farm in Neuendorf, Switzerland. Calvaro's health rapidly declined and he died in September 2003.

Cryozootech founder, Eric Palmer, approached Melliger with the idea of cloning Calvaro in 2002, but because the process was so new Melliger declined Palmer's offer. After the first horse clone was born in 2003, Melliger allowed Cryozootech to take a biopsy, just a few days shy of Calvaro's death.

The first embryos of Calvaro were obtained in 2004, and a first pregnancy was obtained in 2005, however the foal was born premature and weak in 2006 and died of acute septicemia (infection of the blood). A new attempt occurred in 2007, with the resulting foal born healthy this summer. In 2004 Cryozootech started selling shares of the stallion clone for co-ownership, and, to date, has sold 25% of the shares to investors and still owns the remaining shares of the foal, called Calvaro-Cryozootech-Stallion.

Cryozootech has already produced clones of other renowned horses, such as Pieraz, Quidam de Revel, Poetin, Chellano, and, most recently, Gem Twist. Palmer founded the private company in 2001 in order to build an equine cloning research programg. Cryozootech aims to produce its own stallions, which are clones of famous castrated champions. Financial support results from the sale of cloning as a service to owners of stallions, mares, or geldings.​
 

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A clone of champion show jumper Gem Twist is the newest addition to the stable of horses cloned by Cryozootech, a French company that banks equine genetic material. The company announced the clone's birth Sept. 15.

Gem Twist, a grey Thoroughbred gelding, was named "Horse of the Year" three times, won two silver medals at the Seoul Olympic Games, and was named "World's Best Horse" at the 1990 World Equestrian Games. His riders included Greg Best, Leslie Howard, and Laura Chapot. He was retired at New York's Madison Square Garden on Nov. 1, 1997.

As the original Gem Twist was gelded at an early age, his genetic material was never passed on. However, as a genetic replica, the clone will allow the champion's exact genes to enter the breeding pool.
Cryozootech Founder Eric Palmer proposed the idea of cloning Gem Twist to the horse's owners, Frank and Mary Chapot, in 2006. They immediately approved the project.

Gem Twist was euthanized Nov. 18, 2006, at age 27. He died at the site of his birth, the Chapot family farm in New Jersey.​
 

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American Horse Cloning Project Successful

America's first cloned horse turned six weeks old today, according to Texas A&M University (TAMU) researchers who partnered on the successful cloning venture with French scientist Dr. Eric Palmer of Cryozootech. The colt, named "Paris Texas," was produced from skin cells of a European performance stallion, and the active colt is healthy and steadily growing at TAMU.
Katrin Hinrichs, DVM, PhD, Dipl. ACT, professor in the College of Veterinary Medicine at TAMU, led the cloning team on the project. She described Paris Texas as follows: "He's bay with a big white blaze, beautiful eyes, and four white stockings. He's real forward, a real nice foal. Everything is completely normal about him.

"Dr. Palmer took a small piece of skin from the donor animal and grew the cells up in culture and froze them, and he shipped them to us," said Hinrichs. "We did the cloning procedure here, cultured the embryo, and transferred it to one of our recipient mares, who foaled here in the hospital."

The owner of the donor horse from which skin cells were used to produce the clone wishes to remain anonymous. Cryozootech has cells from a variety of high-performance horses in Europe. Since researchers at TAMU were already doing the cloning research, a natural partnership formed between the French company and TAMU to complete this project. Paris Texas is the fourth equine clone to be born in North America, but he is the first horse foal (the other three were mules) and the first to be cloned from adult cells in North America. The mule clones were produced from cells from a fetus.

The TAMU team does all of its cloning work in vitro, or in the laboratory. Rather than getting oocytes (eggs) from mares right before they ovulate (the procedure used to produce the mule clones), TAMU researchers harvest oocytes from mares at other points in the cycle and mature them in an incubator. The scientists then perform nuclear transfers, in which they remove the nucleus from an egg cell (containing the cell's genetic material) and place a donor cell into the enucleated egg. The eggs are then activated, or stimulated to start dividing to form an embryo. Resulting cloned embryos are cultured in the laboratory for seven days, and once they are deemed ready, the scientists transfer them nonsurgically into the recipient mare's uterus just as they would in normal embryo transfer procedures.

About 400 oocytes were cultured during the TAMU project, and the cloning process produced six embryos. Only one pregnancy resulted, and this was carried to term. The grade mare that carried the clone (Greta) has been a part of the reproduction herd at TAMU for about four years.

The colt will be used as breeding stock in Europe. "We're looking for a way to save valuable genetics," said Hinrichs. But cloned foals are unlikely to be competitors. "There are so many variables in the environment that a cloned foal has-Paris Texas was in a petri dish in an incubator for the first seven days of his life," she added. "He was small at birth--about 60 pounds. Sometimes smaller foals don't achieve the adult proportions that they would have. But this horse, genetics-wise, should produce exactly the same sperm and should be able to sire foals with the same genetic makeup as the donor animal."

TAMU has several pregnancies from cloned embryos that are currently gestating and due for arrival in 2006. All were produced using donor cells from American horses. Hinrichs said, "We're so excited about what we're learning about the horse oocyte. We're looking at different methods for treating the donor cells before the nuclear transfer procedure and activating the egg afterward, looking at more ways to build on our results of the study that produced Paris Texas."​
 

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