Fertility in artificially inseminated dairy cattle has declined as milk production has increased, and is one of the most costly challenges faced by today's dairy farmer. This fertility decline causes economic losses of <$300 million annually, losses due in part to damage incurred by sperm during the handling, cooling, freezing and thawing necessary for artificial insemination. For it is the quality of the gamete that is the most important factor in defining fertility (Aitken et al. 2012) and freezing damages the DNA, acrosome and membranes of sperm cells reducing sperm quality and therefore fertility. Membrane Protective Technologies Inc's GameteGuard technology provides protection to sperm cells during cooling, freezing and thawing. In fact, GameteGuard reduces DNA, membrane and acrosome damage by an average of 15-30% in frozen thawed/bovine sperm. More importantly, Phase I breeding trial of 423 lactating cows using two bulls resulted in a 25% improvement in pregnancy rate (P>0.05).The improvements imparted by GameteGuard will have a very real economic impact at two levels of the artificial insemination industry: first, genetics centers (i.e. bull studs or locations where sperm doses are processed) and second, dairy farms. At bull studs ~10% of all processed sperm doses are discarded due to poor motility, DNA, membrane and/or acrosome quality resulting in an estimated annual loss of $3 mm in potential profit per large bull stud per year. At the dairy farm, a 5 percentage point increase in pregnancy rate will add $100 gain per dairy cow per insemination (Overton 2005).A larger breeding trial, the subject of this proposal, is required to statistically confirm an improvement in pregnancy per AI using GameteGuard treated sperm. It is anticipated the addition of GameteGuard to sperm prior to cooling, freezing and thawing will result in commercially relevant increase in pregnancy rates over conventionally frozen sperm due to improved post-thaw sperm health. This breeding trial will lay the groundwork for commercialization in Phase III. The second part of this project is development of a commercial scale quality control program enabling the company to successfully transition to commercial production. The use of GameteGuard at the bull stud will provide a cost effective option to start improving fertility. The integration of this technology at the stud is estimated to cost $0.5-1.00/dose (straw), a cost easily covered by slight increase in straw price which dairy farmers have indicated would be accepted. Interviews revealed they are willing to pay an extra $2-5/straw for increased pregnancy. The respective benefits imparted by the use of GameteGuard will impart a competitive advantage to the U.S. Cattle industry (dairy and beef). Moreover, improving fertility will help enable food production for a burgeoning world population and meets USDA's REE-Action plan objective 1.Membrane Protective Technologies Inc. is poised to commercialize this technology in Phase III of this SBIR proposal. The Phase I results demonstrating efficacy of GameteGuard on bovine sperm, while a sound business on its own, is serving as a model platform upon which applications are being built for other agriculturally important applications including sex-sorted sperm, equine or porcine artificial insemination. This technology is part of an ongoing effort by the USDA's Research, Education, and Economics Action Plan to sustainably intensify agricultural production. By improving fertility using organic extracts such as GameteGuard, dairy farmers will be able to improve their production economics.