Brief Summary of Minutes of Annual Meeting

Brief Summary of Minutes of Annual Meeting: The Chair, Kim Vonnahme called the meeting to order at 8:00 am on Monday June 17, 2013. Kim asked for attendees to introduce themselves. Tom Geary also mentioned students from Fort Keogh who were attending. Tom Geary discussed the meeting activities. A $35 per person fee for the lunch and dinner was announced and that on Tuesday morning (6-18-13), a tour of Ft. Keogh was planned. Kim mentioned the WSASAS meeting that started on 6-19-13 and requested that each researcher provide succinct station reports to ensure sufficient time for the tour and collaboration discussions on 6-18-13. Kim asked for nominations for the 2014 meeting, which included Reno, Fargo, Fairbanks, Las Vegas, and Texas. It was approved to vote on the 2014 meeting location after station reports were completed. Kim announced that Ryan Ashley would be Chair next year. Kristin Govoni was appointed as the new Member at Large. Milan commented that we need our impact statements for the W2112 on how our research and collaborations have impacted the people we serve not necessarily our own research. Milan asked us to provide potentially 6 impact statements reflecting how the W2112 group has supported and enhanced our clientele. Milan commented he would meet with Kim and Ryan to discuss the report. Kim called Adele Turzillo for her report from Washington USDA headquarters. Adele commented she is in a new position (National Program Leader for Animal Production Systems, was appointed Director, Division of Animal Systems, effective April 8, 2013) and that she had emailed her documents to the group, which she then briefly summarized. Dr. Sonny Ramaswamy is the current Director of NIFA (appointed May 7, 2012). Dr. Deborah Sheely is the Assistant Director of the Institute of Food Production and Sustainability. NIFA had a ~18% cut to its programs as listed in Adeles Cornerstone report. HATCH funds should be restored back to 2012 levels. AFRI funds (flagship grant program) should get increase for 2014. Adele commented on The National Association for the Advancement of Animal Science that was formed. On May 17th an amendment was added to farm bill that establishes a new initiative (i.e., farm animal initiative agreement) to address critical needs and research for food security, relationships between human and animals. Adele commented several AFRI panels have met and basically waiting on final fund numbers to come in from budget office before they announce grants funded. Adele added that for Competitive programs, the maximum indirect cost rate (IDC) increased from 22-33% of total costs beginning FY 2012. Adele mentioned other funding opportunities. The Dual Purpose Dual Benefit in Biomedicine and Agriculture Using Agriculturally Important Domestic Species  NIH and NIFA have extended this joint program for an additional 3 years as PAR-13-204. The Ecology and Evolution of Infectious Disease - Funding opportunity partnering NIFA, NSF, NIH, and the U.K. The remainder of the afternoon was dedicated to station reports for Wyoming, Idaho, Texas, Montana, UConn, Illinois, Missouri, Ohio, Minnesota, Nebraska, New Mexico, North Dakota and Mississippi. Voted on 2014 meeting location and dates. The majority approved the 2014 W2112 meeting to be held June 10th and 11th in Fairbanks, Alaska. Kim Vonnahme adjourned meeting at 5:00 pm. Tuesday, 6-18-13 Kim Vonnahme called the meeting to order at 7:30 am. Tom Geary provided a brief PowerPoint presentation on the history of Fort Keogh. A tour of a portion of the barns and facilities of Ft. Keogh followed. Collaborative projects for 2013-2014 were discussed. Kim commented she had sent an email to the group with previous collaborations. We agreed to modify and send changes to Kim and Ryan to include in the report. Milan commented we have 90 days to submit the report and we need to focus on providing ~6 impact statements that we could include. It was discussed among the group to put together a list and send to Ryan describing how we are achieving the milestones listed in the project. Specifically, presentations to various producers are needed so we can document how we have and continue to serve our clientele. Kim and Ryan will combine from all the W2112 members and include in the report. The rest of the morning focused on developing our Impact Statements to include in the report as well as some of the outputs from the W2112 project. Specific outputs discussed were: 1) Reviewed current status of estrous synchronization handbook by end of 2013. Will put on websites (e.g., Ag Experiment Station Websites). 2) Efforts underway to identify speakers to talk at next ruminant BIF. 3) Maybe do a review (JAS) about fetal programming based on work from the W2112 group. Include a history of W2112. This will probably be discussed next year. 4) Kim will get in touch with Teglas about the milestone concerning the vaccine (this was one of the milestones listed). Allen Bridges suggested putting together the estrous synch manual and distribute through BIF. Tom Geary said he will contact BIF and Allen offered to help. The group discussed the possibility of building of a series of webinars and doing this through ASAS. Kim and Andrea said they would contact Megan at ASAS regarding the webinars, which might fit with ASAS. Symposia suggestions for ASAS were also discussed. A fetal programming symposium was suggested as many in the W2112 have research related. Attendees were encouraged to think of other areas that W2112 could address to use as symposia suggestions. These will probably be included in our next submission for the W2112 project submitted January of 2016. It was suggested to appoint a re-write committee next year in Fairbanks. Potential Impacts were discussed. Kim asked for small groups to get together and draft impact statements. A number of drafts were considered and many in the group provided hard copies and/or emailed to Kim and Ryan for inclusion in the report. We narrowed the statements down to the following with the understanding that edits will be made before inclusion into the final report: 1) Demonstrated that with management and genetic selection, Bos indicus females are capable of calving at 2 years of age versus 3 years of age. This will save ~$500 per heifer for the producer and increases life time economic efficiency 6-10%. 2) Prenatal stress (stress to the pregnant female) reduces profitability of the calves due to increased temperamental dispositions and reduced response to vaccination. The value of these calves is reduced by at least 10% from the value of unstressed calves. 3) Collaborative efforts allowed rapid treatment of spermatozoa with magnetic nano-particles to remove defective spermatozoa and allow equal and high pregnancy rates with 50% reduction of the sperm dose used with artificial insemination in cattle. This allows production of twice the number of semen doses per ejaculate for dissemination of superior genetics. This effort also impacts human assisted reproductive therapies, wherein sperm quality biomarkers can be used for accurate diagnosis of male infertility and clinician decision making. 4) Multistate, institutional, and investigator approach towards the reduction in average date of conception rate in the breeding season by 10-15% in beef cattle through the development of improved estrous synchronization approaches that has increased the reproductive efficiency and production profitability of beef cattle production systems. 5) Enhanced the understanding of how phenotypical, physiology, and endocrine characteristics, including antral follicle count, growth rate and efficiency, follicular growth patterns, stress response, and estrous expression impacts on reproductive competence. 6) Determined influence of endocrine environment during follicular development on oocyte competence to translate to improved methods of estrous synchronization, multiple ovulation protocols, and in vitro embryo production. Collaborative projects for 2013-2014 were discussed. Collaboration Between Stations. Project members meet annually to present results of completed studies, review the progress of on-going work, discuss industry issues and plan and coordinate the initiation of new studies for the upcoming year(s). We have created a network of collaborating scientists focused on improving the fertility of domestic ruminants in the West. The founding members of the W-2112 Regional Research Project established a tradition of cooperation that the current members strive to continue. Indeed, in this era of instant and direct communication, cooperation and collaboration has never been greater or more significant. The extent of collaboration between stations is extensive and may best be described by providing examples of collaborative projects. Collaborative projects in brief include: Determining hormones patterns in ewe lambs with differing residual feed intakes (RFIs; NM, MT); Effects of nutrient restriction on metabolic hormone patterns (NM, CO); Thyroid involvement in BVD infected cows (NM, CO); Metabolic hormone profiles in cows consuming endophyte infected fescue (NM, AR); Follicular fluid concentrations of IGF (NM, OH), Meta-analysis of birth type on ewe reproductive performance (WY, MT, NM); ISG concentrations on maternal blood and IFNt in conceptuses (CO, OH); ISG15 knock out mice to understand the role of ISG15 in the bovine uterus (CO, WA); PGEs and CL function using intraluteal implants (CO, HI, TX); Influence of progesterone on sexual behavior (AK, WY, MN, MT, NE); Influence of gestational nutrition on pancreatic function (AZ, CO); Influence of gestational nutrition on insulin (AZ, WY); Extended oocyte/embryo culture (MI, FL); Oocyte quality and follicle diameter (MI, MT); Influence of follicle age on fertility (OH, MT); Stress hormones and abortion in cattle and sheep (NV, TX); Inhibin concentrations and temperament (TX, MN); Development in intact males (WY, MT, NM); SNPs in sheep (WY, UT); Role of VEGF in spermatogonial stem cell development and differentiation (WA, NE); Evaluation of IFN in maternal blood and embryo culture media in relation to early pregnancy establishment using a sensitive radioimmunoassay (CSU & VT); Uterine capacity for pregnancy in cattle (WSU & UFL with Peter Hansen [not a W2112 member]); Role of sperm abnormalities in pregnancy success and embryonic mortality in cattle (MT, MO); Examination of sperm quality in prenatally stressed compared with nonstressed control Brahman bulls (TX, MO); Effects of endogenous steroid levels during follicular development and ovulatory follicle size on oocyte quality/competence and uterine environment (OH, MO, MN, UID, & MSU); Optimizing approaches for estrous synchronization and timed-AI in beef cattle (OSU, ISU - Patrick Gunn [non-W2112], UW - Scott Lake [non-W2112]). Androstenedione (A4) concentrations and clearance in livers of high A4 and low A4 cows (MS, NE); VEGF isoforms in bull testis (NE, OSU); CXCL12/CXCR4 signaling and VEGF involvement (NE, NM); Muscle, adipose, and bone studies using obese models (NE, UConn); RNA extraction techniques from adipose tissue (NM, UConn); Poor maternal nutrition affects postnatal growth and development of lambs (UConn, ND); Evaluation of enzymes responsible for progesterone metabolism in ewes treated with hCG (NM, MS); Other examples of cooperation include co-advising of graduate students between stations and willingness of member scientists to freely share critical reagents and methodologies with other members of the regional project. Such reagents include antisera, vaccines, tissue and serum samples, RNA and DNA tissue libraries, and novel cell lines and cDNA probes. Kim adjourned the 2013 W2112 meeting at 11:00 am. Producer directed output  1. Alexander B. and Gary E. Moss. 2012. BREEDING PERFORMANCE OF RAMS: 25% of rams sire nearly 40% of the lambs. UW College of Agriculture and Natural Resources. Reflections. p 20-21. 2. Alexander B. Producer oriented seminar. Effect of Sex of Co-Twin and Breed on Ewe Flock Productivity. National America Sheep Industry Meetings, Nashville, TN. January 21, 2010. 3. Alexander B. Producer oriented seminar. Raising Sheep. UW Extension, Park County. January 27, 2011. 4. Alexander, B. University of Wyoming Study Reveals Robust Sheep Industry. University of Wyoming Extension News http://www.wyomingextension.org/news/2012/05/23/university-of-wyoming-study-reveals-robust-wyoming-sheep-industry/ and Wyoming Public Radio audio report. 5. Bridges, A. Producer oriented seminar. Applied Reproductive Strategies in Beef Cattle Symposium (Sioux Falls, SD) - December 2012 -  Changes in body condition on oocyte quality and embryo survival - ~ 200 producers/professionals. 6. Bridges, A. Producer oriented seminar. Driftless Region Beef Conference (Dubuque, IA) - February 2013 - Replacement female strategies - ~100 producers. 7. Bridges, A. Producer oriented seminar. Form-A-Feed Spring Breeding Webinar - May 2013 - Preparing for the breeding season - ~30 producers. 8. Bridges, A. Producer oriented seminar. Minnesota Beef Showcase - March 2012 - It takes two to tango: Importance of bull management - ~35 producers. 9. Bridges, A. Producer oriented seminar. Minnesota Beef Showcase - March 2012 - Options for estrous synchronization - ~35 producers. 10. Bridges, A. Producer oriented seminar. Minnesota Cow/Calf Days (10 locations) - February 2013 - Incorporating reproductive technologies into the cow herd - ~450 producers/professionals. 11. Bridges, A. Producer oriented seminar. Minnesota Nutrition Conference - Septermber 2012 - Influence of pre- and post-AI nutrition on pregnancy success in beef cattle - ~40 producers/professionals. 12. Bridges, A. Producer oriented seminar. Northern States Beef Conference (Watertown, SD) - January 2013 - Economic and managerial considerations when deciding to develop or purchase replacement females - ~125 producers/beef industry professionals. 13. Bridges, A. Producer oriented seminar. Northwest MN Cattlemens Association Meeting - October 2012 - Preparing you beef operation for winter - ~45 producers. 14. Bridges, A. Producer oriented seminar. University of Minnesota Cattle Harvest Days - January 2012 - Genetic selection and use of genomics to improve carcass quality - ~50 producers/beef industry professionals. 15. Bridges, A. Producer oriented seminar. University of Minnesota Cow/Calf Days (5 locations) - February 2012 - Replacement heifer development ~250 producers. 16. Bridges, A. Producer oriented seminar. XVII Course on New Approaches to Production and Reproduction in Cattle. March 14-15, 2013, Uberlandia, Brazil. - March 2013 - Uterine function and embryo survival - ~1000 producers/professionals. 17. Bridges, A. Producer oriented seminar. XVII Course on New Approaches to Production and Reproduction in Cattle. March 14-15, 2013, Uberlandia, Brazil. - March 2013 - Progesterone before AI and after AI on pregnancy success - ~1000 producers/professionals. 18. Burdick Sanchez, N.C., B.L. Bradbury, J.A. Carroll, R.C. Vann, T.H. Welsh, JR and R.D. Randel. 2012. Metabolic differences in temperamental Brahman cattle can affect productivity. Proc Beef Improvement Federation (BIF) 2012 pp 94-146. 19. Burton, Devin J., Paul A. Ludden, Robert H. Stobart, Brenda M. Alexander. Fifty years of the Wyoming Ram Test: How have we changed? The Shepard. 57 (2):6, April 2012. 20. Cardoso, FC. Producer oriented seminar. 2013. Managing the Transition Cow. Illinois Dairy Summit. January 22-24, Bloomington, Centralia, and Freeport, IL. Role: conference organizer and presenter. Audience: dairy producers, dairy industry representatives, and consultants. 314 attendees. 21. Cardoso, FC. Producer oriented seminar. 2013. The 3-R Transition Period: Recovery, Reproduction, and Results. Four State Dairy Nutrition and Management Conference. June 12-13, 2013, Dubuque, IA. Role: conference organizer and presenter. Audience: dairy producers, dairy industry representatives, and consultants. 529 attendees. 22. Cruppe L. H. and M. L. Day. 2011. Producer oriented seminar. Estrous synchronization protocols to optimize follicular dynamics and hormonal concentration in Brazil and the USA. The XV Course on New Approaches to Production and Reproduction in Cattle. Uberlandia, Brazil, March 17-18, 2011. Presentation to approximately 700 producers, veterinarians, students and allied industry. 23. Cruppe L. H. and M. L. Day. 2011. Producer oriented seminar. Maximizing timed AI pregnancy rates with the 5 day CO-Synch + CIDR protocol in beef cattle. The 9th IRAC International Symposium on Animal Reproduction. Cordoba, Argentina, September 9-11, 2011. Presentation to approximately 1000 veterinarians, students and allied industry. 24. Cupp, AS, JR Wood, RA McFee, R Slattery, KA Beavers, W Pohlmeier, K Sargent NX Lu, J Smith, J Kerl, Vanessa Brauer, A Summers, Stetson Weber, RA Cushman. Granulosa Cell Gene Expression is altered in Follicles from Cows with Differing Reproductive Longevity. Nebraska Beef Report 2011. 25. Cupp, AS, Producer oriented seminar. 2012- Nebraska Ag Builders- Research conducted at the Agriculture Research and Development Center in the cow/calf unit Physiology herdPotential effects of too much androgen on female fertility. 26. Gardiner, Lane, Benjamin Rashford, John Hewlett, Brenda Alexander. 2012. The State of the Wyoming Sheep Industry. University of Wyoming Extension Bulletin. B-1229. 27. Geary, T. Producer oriented seminar. Genex Customer Meeting in Lewistown, MT (1/ 2012), Reproduction and synchronization in beef cattle (approximately 70 producers). 28. Geary, T. Producer oriented seminar. Genex Large Herd Summit in Billings, MT (9/2012), Bull fertility measures: Impacts on fertilization and pregnancy success (approximately 50 producers from across US, each of which breeds > 2500 cows). 29. Geary, T. Producer oriented seminar. MT Extension group in Big Timber, MT (1/2012), Fertility and pregnancy establishment in beef cattle (approximately 40 producers). 30. Geary, T. Producer oriented seminar. MT Extension group in Glendive, MT (1/2013), Bull fertility measures and their impacts on fertilization and pregnancy success (approximately 40 producers). 31. Geary, T. Producer oriented seminar. MT Nutrition Conference in Bozeman, MT (4/2012), Fertility in beef cattle (approximately 150 producers & industry representatives). 32. Geary, T. Producer oriented seminar. MT Producer group in Miles City, MT (4/2013), Fertilization and pregnancy maintenance: What is new? (approximately 30 producers). 33. Geary, T. Producer oriented seminar. MT Producer group in Miles City, MT (9/2012), Fertilization and pregnancy maintenance: What is new? (approximately 30 producers & industry representatives). 34. Geary, T. Producer oriented seminar. Pfizer Scientific Exchange in Miles City, MT (5/2012), Fertilization and pregnancy maintenance in the beef cow: Ovulatory follicle size & estradiol (approximately 15 industry veterinarians). 35. Geary, T. Producer oriented seminar. Pfizer Scientific Exchange in Miles City, MT (5/2012), Bull fertility measures and their impacts on fertilization and pregnancy success (approximately 15 industry veterinarians). 36. L. H. Cruppe and M. L. Day. 2012. Producer oriented seminar. Optimizing timed AI pregnancy rate with the 5-d CO-Synch + CIDR Program. The 2012 Joint Convention of CETA/ACTE and AETA (Canadian and American Embryo Transfer Associations). Presentation to approximately 300 members of these societies. 37. Lu NX, J Smith V Brauer, A Summers, W Pohlmeier, KA Beavers, R McFee, K Sargent J Kerl, RA Cushman, AS Cupp, JR Wood. Ooctye mRNA and Follicle Androgen Levels associated with Fertility. Nebraska Beef Report 2012, Pg 28-29. 38. M. L. Day. 2012. Producer oriented seminar. Optimizing timed AI pregnancy rate with the 5-d CO-Synch + CIDR Programme. The 2012 Bayer New Zealand Reproduction Road Show. Presented at 8 locations in New Zealand to a total of approximately 300 dairy and beef cattle veterinarians. 39. Maquivar M, and M. L. Day. 2011. Producer oriented seminar. Strategies for nutritional and hormonal induction of puberty in beef heifers and the impact on fertility. The 9th International Symposium on Animal Reproduction. Cordoba, Argentina, September 9-11, 2011. Presentation to approximately 1000 veterinarians, students and allied industry. 40. Maquivar M. and M. L. Day. 2011. Producer oriented seminar. Nutritional and hormonal induction of puberty in heifers and the impact on fertility. The XV Course on New Approaches to Production and Reproduction in Cattle: 43-54. Uberlandia, Brazil, March 17-18, 2011. Presentation to approximately 700 producers, veterinarians, students and allied industry. 41. McRee, RM, RA Artac, WE Pohlmeier, JG Kerl, VM Brauer, RA Cushman, AS Cupp. Vascular Endothelial Growth Factor A (VEGFA) in Ovulatory Follicles. Nebraska Beef Report 2012, pg. 26-27. 42. Randel, R.D., R.C. Vann and T.H. Welsh, Jr. 2012. Selection tools for temperament. Proc. Beef Improvement Federation (BIF), 2012 pp 38-75. 43. Summers AF, RA Cushman, SP Weber, KV Moline, JW Bergman, ML Spangler, AS Cupp. Nutritional Regime and Antral Follicle Count Impact Reproductive Characteristics in Heifers. Nebraska Beef Report 2012, pg. 24-25. 44. Summers, AF, RA Cushman, KV Moline, JW Bergman, AS Cupp. Heifers with low antral follicle counts have low birth weights and produce progeny with low birth weights. Nebraska Beef Report 2013. Pg 11-12. 45. Sutovsky P. Producer oriented seminar. 4th Annual Symposium of Réseau Qébeçois en Reproduction, Montreal, November 15 &16, 2011. 46. Sutovsky P. Producer oriented seminar. 8th Biannual Meeting of the Association for Applied Animal Andrology (AAAA), July 28 & 29, 2012, Vancouver, BC, CANADA. 47. Sutovsky P. Producer oriented seminar. Asian Productivity Organization (APO) Workshop on Nanotechnology Applications for Boosting Agricultural Productivity, Yilan, Taiwan, R.O.C., September 3-7, 2012 (Invited speaker & facilitator/resource person) 48. Sutovsky P. Producer oriented seminar. From Researcher to User - Workshop on Knowledge Transfer, Réseau Qébeçois en Reproduction, Montreal, November 14, 2011. 49. Sutovsky P. Producer oriented seminar. International Symposium on Animal Reproduction and Genetics, Lima, Peru, August 18, 2011. 50. Sutovsky P. Producer oriented seminar. Kansas City Animal Health Investment Forum, August 29-30, 2011. 51. Sutovsky P. Producer oriented seminar. Reproductive Strategies in Beef Cattle, Joplin, MO, August 31-September 1, 2011.

Accomplishments: Objective 1. GnRH agonist treatment results in the following: 1. Increases CL size and circulating P4 concentrations 2. Increases number of antral follicles in the first follicular wave 3. Cease follicular waves after the first wave in first lactation cows, but multiparous cows continue to have greater number of large follicles than control animals 4. Greater numbers of follicles on the ovary with the CL indicating a potential intraovarian communication 5. No change in interestrous length if the animal is not pregnant 6. Increased pregnancy rates in cows receiving agonist for 7 days as compared to 12 days of treatment or controls 7. No difference in E2 or P4 concentrations in milk 8. No residual Deslorelin measured within the milk Impacts of nutrient restriction followed by realimentation on uterine blood flow and placental vascular reactivity in beef cows. 1. The uteroplacenta appears to respond differently to nutrient restriction in pregnant beef cows compared to sheep. 2. The down-regulation of hepatic cytochrome P450 activity during pregnancy may contribute to the increase in peripheral estradiol concentrations in beef cows. 3. Chronic uterine infusion of melatonin increases fetal aorta blood flow and umbilical artery blood flow, while melatonin receptor antagonist (luzindole) infusion decreases fetal aorta blood flow in sheep.. Influence of post-insemination nutrition on embryonic development in beef heifers and influence of estrus at fixed-time AI on accessory sperm numbers and embryonic development. 1. Immediate alterations in early embryonic development are observed in heifers that fail to receive adequate nutritional inputs following insemination and these alterations are likely due to insufficient oviduct and uterine support of the developing embryo. 2. Estrus expression prior to timed-AI resulted in day 6 embryos of greater quality and developmental stage compared to embryos recovered from cows that failed to exhibit estrous prior to timed-AI. In addition, the number of accessory sperm tended to be greater; potentially indicating improved sperm transport in cows that exhibited estrus. 3. Nutritional and hormonal interventions have shown promise to advance sexual maturation in bulls. Technology to decrease the age at which viable sperm is produced by bulls intended for AI in the genomics era of sire selection is necessary to capture the full advantages of genomics to accelerate genetic improvement. Determining if a dose dependent response to the etiologic agent that causes Epizootic Bovine Abortion (aoEBA) is occurring in animals that have been exposed to the pathogen prior to pregnancy and determine if bacterial load prior to pregnancy can have an effect on maintenance of pregnancy. 1. Investigation into the role of immunologic mediators during pregnancy following infection with the abortigenic bacteria that causes Epizootic Bovine Abortion may ultimately lead to a better understanding of the role of cytokines in the maintenance and termination of pregnancy in cattle infected with a bacterial pathogen. 2. Research into the development of a safe vaccine, one that is non-pathogenic to the host and can be used in pregnant animals, will be aided by the results of this research. Understanding which immunologic mediators play a role in a protective immune response against this pathogen will help tailor research into the development of a possible recombinant vaccine that could serve this purpose. 1. IFNT protects early pregnancy in ruminants through paracrine action on the endometrium and also through endocrine action on the CL and peripheral blood cells through enhancing luteal resistance to PGF.2. A highly specific radioimmunoassay has been developed for IFNT, which has different levels of detection in uterine flushing compared to serum. 3. The endocrine mechanism of action of IFNT might entail up regulation of ISGs as well as cell survival genes to assist with providing resistance of the CL to PGF, but also in longer-term resilience in context of continued production of progesterone.4. IFNT may also induce luteal resistance to PGF through induction of ISGs that prevent downregulation of genes encoding LHCGR and PTX3. Stabilization of LHCGR would support steroidogenesis and continued production of P4, whereas stabilization of PTX3 may prevent cell death and mediate inflammatory responses in a humoral rather than cell-mediated mechanism.5. The direct assessment of gene function within the placenta has not been feasible in non-rodent species, especially in ruminants. A good case in point is that the placental lactogen has investigated for nearly 50 years, yet a direct function or relative importance has never been established. 6. The quantification of changes in the estrogen-kisspeptin-GnRH signaling pathway in OVX-E ewes at different times of the year may lead to insights into factors causing the switch from negative to positive. A Bovine Model for Polycystic Ovary Syndrome. 1. A sub-population of cows within the UNL physiology research herd have androstenedione (A4) excess within follicular fluid. These cows appear to leave the herd at a 2:1 ratio compared to cows that have low A4 due to not having a calf. Granulosa cells activate different signal transduction pathways dependent on follicle health status and ability to convert androstenedione to estrogen resulting in different steroidogenic profiles for beef cattle follicles. 1. We are investigating cows that have been treated with different progesterone or progestin as well as those with different concentrations of androgen in follicular fluid to understand how signal transduction pathways in granulosa cells may be altered. Prostaglandin F2a Activates Stress Response Signaling and Induces Expression of Activating Transcription Factor 3 (ATF3) in Bovine Large Luteal Cells. 1. Activating transcription factor 3 (ATF3) is a stress-adaptive gene that regulates proliferation or apoptosis under stress conditions. It appears to be upregulated during CL lysis initiated by PGF2alpha. This gene appears to suppress transcription of genes however, it does not appear to affect steroidogenic enzymes but does suppress progesterone production. Loss of Vascular Endothelial Growth Factor A (VEGFA) isoforms in Granulosa cells using pDMRT-1 cre reduces female fertility by arresting follicular development and reducing number of follicles ovulated and affecting litter size. 1. Novel mouse lines where VEGFA isoforms have been knocked out in granulosa or Sertoli cells have been generated to delineate the functions and necessity for VEGFA isoforms in ovarian function. The actions of these VEGFA isoforms appear to have dramatic impacts on female reproduction. Loss of all VEGFA isoforms in Granulosa and some germ cells causes reduced fertility with smaller litter sizes and more days from mating to parturition. There are also less antral follicles developing with increases in mRNA abundance for FOXO3a, reduced estrogen concentrations in plasma, and smaller ovarian weight with a tendency for less CL. These all suggest that loss of all VEGFA isoforms causes subfertility by causing altered follicular development resulting in less follicles ovulating and smaller litters. Poor maternal nutrition affects postnatal growth and development of lambs. 1. Poor maternal nutrition due to over-feeding during gestation leads to increased size of lambs at birth, increased heart size and increased insulin which may predispose these offspring to metabolic disorders later in life.2. Poor maternal nutrition due to under- and over-feeding during gestation impairs muscle development during postnatal growth. The reduced muscle growth may be due to altered gene expression within the muscle and satellite cell number and/or function.3. Poor maternal nutrition during gestation alters the expression of key genes involved in the ability of mesenchymal stem cells to differentiate into adipocytes and osteoblasts. This could explain the differences in bone and adipose tissue development between offspring from control versus under- or over-fed mothers during gestation.4. Poor maternal nutrition during gestation alters circulating concentrations of GH, IGF-I, insulin and leptin, which may contribute to, altered growth observed in offspring from these mothers. Expression profiling and potential functions of protein kinase C delta during early bovine embryo development 1. Recent work implicates PKC-delta as a mediator of embryonic development, interferon-tau expression and early placental development in cattle. Development of a Luciferase-Based Reporter Assay for Quantifying Interferon Activity 1. An ISRE-reporter assay was developed as a replacement for the virus-dependent cytopathic assays that have been used to examine interferon-tau in biological samples. This assay contains a similar sensitivity, a broader concentration dose-response range, and is completed in a shorter time than the traditional cytopathic assay. Correlations Between PAG Concentrations, Pregnancy Loss and Milk Production in High Producing Holstein Cows 1. An initial assessment of a large database determined that placental and luteal insufficiencies are detected in dairy cattle prior to pregnancy failure. Continued analysis of this dataset will explore the impact of peri- and post-partum illnesses on pregnancy outcomes in dairy cattle. Impact of the number of estrous cycles before the start of breeding on reproductive performance of beef heifers and effects on subsequent re-breeding success. Attainment of puberty before start of breeding was associated with increased heifer pregnancy rate, with little or no benefit from having more than one estrous cycle prior to start of breeding. In contrast, second season pregnancy rate was positively associated with number of estrous cycles expressed prior to the start of a heifer's first breeding season. Circulating bovine pregnancy associated glycoproteins (bPAGs) are associated with late embryonic/fetal survival but not ovulatory follicle size in suckled beef cows No relationship exists between serum bPAGs and ovulatory follicle size or embryo stage/quality at ET; however, cows that lost an embryo after d 28 had lower concentrations of bPAGs on d 28 compared to cows that maintained pregnancy. Activation of the CXCL12/CXCR4 chemokine axis may drive vascularization of the ovine placenta The CXCL12/CXCR4 signaling pathway is activated during implantation and placentation in sheep and is likely playing a role in the communication between trophoblast cells and the maternal endometrium, specifically driving vascularization. Effects of maternal obesity on membrane progesterone receptors, CXCL12, and CXCR4 expression during early pregnancy in sheep Membrane progesterone receptors are up regulated in caruncle tissue from obese compared to control ewes. PGE1 and PGE2 functions in rats and sheep An essential fatty acid deficiency increased PSP and decreased the deciduomata response in PSP rats. Both PGE1 and PGE2 increased in pregnant ewes support PGE1 and PGE2 as antiluteolysins. PGE1 or PGE2 prevents progesterone-induced premature luteolysis by preventing a loss of luteal unoccupied and occupied LH receptors. Maternal obesity and overnutrition is associated with reduced systemic progesterone during the estrous cycles of adult female offspring and a marked increase in insulin resistance at midpregnancy Mature ewes born to obese/overfed mothers (F1-MO ewes) exhibit marked greater insulin resistance with elevated blood concentrations of glucose and insulin throughout pregnancy when compared to daughters born to control fed mothers even when both groups were only fed to requirements throughout pregnancy. These data suggest that the F2 generation born to these F1-MO ewes may exhibit the same metabolic alterations (hyperphagia, increased adiposity, and increased insulin resistance) as their mothers, resulting in a transgenerational programming effect. Dexamethasone (DM) administration in F0 ovine pregnancy eliminates the postnatal (PN) plasma leptin (L) peak in F2 offspring (OFF) and results in increased appetite and weight gain These data confirm that injection of a synthetic glucocorticoid to pregnant ewes in late gestation eliminates the normally observed postnatal leptin surge in their offspring leading to increased appetite and obesity in later life. This confirms our hypothesis that the elevated levels of cortisol observed in blood of fetuses gestated by obese/overfed ewes may be the cause for the elimination of the postnatal leptin surge in these offspring, leading to macrophagia, obesity and insulin resistance in adulthood. Maternal obesity and over-nutrition alters the fetal sheep heart transcriptome We have previously demonstrated that maternal obesity and overnutrition increases fetal left ventricular heart weight and reduces cardiac work load capacity. There is strong evidence that the satiety hormone leptin, produced predominantly by adipocytes has effects on the maturation of the hypothalamus circuits controlling food intake. We have recently confirmed the presence of a leptin surge from day 6 to 9 postnatal age in the sheep, and that diet-induced maternal obesity eliminated this postnatal peak in their daughters. More importantly, when these offspring reached adulthood, and were subjected to ad libitum feeding, they exhibited increased appetites, adiposity and insulin resistance. Obesity in pregnancy: implications for the next generation To evaluate transgenerational impacts of maternal obesity, F1 daughters of obese/overnourished (MO) and control-fed (C) ewes were fed only to requirements during pregnancy. From mid to late gestation, F1 MO mothers exhibited markedly greater baseline glucose and insulin than F1 C mothers, and were more insulin resistant. As observed for their mothers, F2 lambs born to F1 MO ewes had a markedly greater fat % at birth than F1 C mothers, and failed to exhibit the postnatal leptin spike as their mothers had. These data suggest a predisposition of F2 MO offspring for increased appetite, adiposity and metabolic disease in adult life. Late gestational low dose dexamethasone treatment programs arterial hypertension and vascular dysfunction in adult female sheep offspring Administration of exogenous glucocorticoids to pregnant ewes on days 103/104 of gestation programs arterial hypertension in adult female offspring. Dietary intervention in early pregnancy reduces the negative impacts of obesity /overnutrition on fetal growth and organ development, and improves glucose-insulin dynamics in late gestation Reduction in maternal feed intake only to requirements at day 28 of gestation in obese/overfed ewes through late gestation has been shown to normalize fetal growth and glucose:insulin dynamics both at mid- and late gestation. Dietary reduction from early gestation in obese/overnourished ewes reduced adiposity and serum lipids and increased liver glycogen in late gestation fetuse Late gestation fetuses of obese/overfed ewes exhibit increased adiposity, and elevated serum concentrations of triglycerides and cholesterol, as well as an increased PEPCK expression and reduced glycogen in their livers when compared to similar age fetuses from ewes fed only to requirements. Reduction in maternal feed intake only to requirements from day 28 through the rest of gestation in obese/overfed ewes returned all of these values to those of controls by late gestation. Neural Activity in the olfactory pathway of female-, male-oriented, and sexually inactive rams exposed to urine from estrous and ovariectomized ewes Female-oriented rams exhibit more neural activity along the olfactory pathway when exposed to sexually evocative stimuli than do sexually inactive rams. Exposure to sexually evocative and neutral odors elicited differential activity only in cortical amygdala in female-oriented rams. Activity in the cortical amygdala may alert rams and may account for the different behavioral response when sexually active rams are exposed to either estrous or non-estrous females. The lower neural activity in sexually inactive rams suggests inactive rams do not cue to sexual stimuli as sexually active rams do. The presence of appetite hormones in breast milk may be important in infant appetite regulation. Additionally, changes in concentration of GLP-1 and Ghrelin during a single feeding may signal satiety in the nursing infant. Fetal adrenal demedullation at 0.7 gestation alters fetal growth and metabolism at 0.9 gestation in an ovine model of placental insufficiency and intrauterine growth restriction. Norepinephrine is required for normal ß-cell development and function. Chronic excess of NE as found in IUGR fetuses potentiates the actions of hypoglycemia and hypoxemia to suppress insulin secretion and fetal growth. Intrinsic deficiencies in myoblast proliferation result in less differentiated myoblasts and smaller myofibers in IUGR fetal sheep IUGR myoblasts possessed intrinsic functional deficiencies that impaired their capacity to proliferate and facilitate muscle growth. In vitro differentiation of myoblasts resulted in an alter fiber phenotype profile between control and IUGR fetuses. A heterobivalent ligand composes of GLP-1 and adrenergic antagonist, yohimbine, exhibits high specificity for ²-cells in vivo. Multivalent ligands can be expanded and used to develop other ²-cell specific targeting agents or reproductive specific targeting agents. Genetic parameters of three methods of temperament evaluation of beef calves and Evaluation of temperament scoring methods for beef cattle Temperament measured using exit velocity or pen score is heritable (0.27 to 0.49). This is adequate for selection to improve temperament of cattle. If only one method is to be used pen score has the greatest heritability (0.49). Calves can be effectively pen scored as single animals or in groups up to 5 animals. The correlation between single animal and group pen scores was 0.79. Effects of prenatal transportation on Brahman calves Prenatal transportation stress applied on days 60, 80, 100, 120 and 140 of gestation resulted in: a. More temperamental calves on days 14 and 28 after birth. b. Only minor differences in complete blood cell counts due to prenatal stress but significantly (<0.01) greater numbers of basophils and a tendency (P=0.10) for greater numbers of neutrophils in females compared with males. c. Response to vaccination in white blood cell counts was reduced in prenatally stressed heifers but not in bulls. d. Prenatal stress and temperament alter metabolic response to an endotoxin challenge in Brahman bull calves. Circulating glucose was greater in prenatally stressed calves and in temperamental calves. Following endotoxin challenge insulin was lower in prenatally stressed calves. e. Prenatal stress influenced pre and post-endotoxin challenge acute phase response in Brahman bull calves. Rectal temperatures were greater pre and post-endotoxin challenge in prenatally stressed Brahman bull calves. Cows that received controlled energy (CE) diets during the last 3 wk prepartum had shorter days to pregnancy (DTP) than cows that consumed high energy (HE) diets in this time period, which may be attributable in part to increased NELI in the first 4 wk postpartum for cows that received CE diets in the close-up (CU) period. In addition, lower BCS loss in the first 6 wk and slightly greater glucose concentrations at wk 3 may have contributed to improved reproductive performance. Energy- limited cows had lower liver TG concentrations at wk 2, which led to fewer DTP. Objective 2: Effects of progesterone concentration and FSH administration on follicle number and oocyte competence in beef cows and effects of decreased progesterone concentrations during follicular development on oocyte yield and quality in beef heifers. 1. Administration of FSH increased the number of follicles aspirated, oocytes collected, proportion of grade 1 to 3 oocytes, and number of grade 1 to 3 oocytes per cow. Administration of FSH, however, did not affect the ability of the oocyte to cleave or develop into a blastocyst or affect the characteristics of subsequently developed blastocysts. Reducing progesterone concentrations during follicular development increased the number of follicles present at aspiration but did not impact quality of oocytes recovered or the ability of the oocytes recovered to cleave and develop into a blastocyst. 2. In beef heifers, reducing progesterone concentrations within an ultrasound-guided ovum pick-up protocol that included FSH administration resulted in more oocytes collected and increased the number of grade 1 to 3 oocytes recovered per female. Nutritional management of Nelore heifers to support precocious pregnancy at 12 to 15 months of age in Brazil 1. It was demonstrated that a majority of Bos indicus (Nelore) heifers will conceive at one year of age and rebreed effectively as primiparous cows with sufficient nutritional management and hormonal intervention. Impacts of nutrient restriction followed by realimentation on uterine blood flow and placental vascular reactivity in beef cows. 1. The uteroplacenta appears to respond differently to nutrient restriction in pregnant beef cows compared to sheep. Nutrient restriction does NOT alter uterine blood flow in early to mid pregnant beef cows. Upon realimentation, it appears that the ipsilateral horn receives more blood flow compared to cows that were not previously restricted. 2. The down-regulation of hepatic cytochrome P450 activity during pregnancy may contribute to the increase in peripheral estradiol concentrations in beef cows. Hepatic glucuronosyltransferase is down-regulated while serum concentrations of progesterone are increased after extended maternal nutrient restriction. 3. Chronic uterine infusion of melatonin increases fetal aorta blood flow and umbilical artery blood flow, while melatonin receptor antagonist (luzindole) infusion decreases fetal aorta blood flow in sheep. Influence of post-insemination nutrition on embryonic development in beef heifers and influence of estrus at fixed-time AI on accessory sperm numbers and embryonic development. 1. Immediate alterations in early embryonic development are observed in heifers that fail to receive adequate nutritional inputs following insemination and these alterations are likely due to insufficient oviduct and uterine support of the developing embryo. 2. Estrus expression prior to timed-AI resulted in day 6 embryos of greater quality and developmental stage compared to embryos recovered from cows that failed to exhibit estrous prior to timed-AI. In addition, the number of accessory sperm tended to be greater; potentially indicating improved sperm transport in cows that exhibited estrus. The effect of a post-weaning high energy diet on the testicular characteristics and semen production in mature Holstein bulls and Effect of Exogenous FSH on Endogenous FSH Secretion and Testicular Development in Prepubertal Bulls. 1. Nutritional and hormonal interventions have shown promise to advance sexual maturation in bulls. Technology to decrease the age at which viable sperm is produced by bulls intended for AI in the genomics era of sire selection is necessary to capture the full advantages of genomics to accelerate genetic improvement. Linking mutations in genes that control male fertility to normal and aberrant sperm phenotypes in bulls used in artificial insemination service. Variations in the genome to sperm protein phenotype can be linked to objectively assess phenotypic expression of polymorphic fertility-associated genes. Many elite bulls with outstanding meat or milk production traits have low AI fertility. Based on sperm quality biomarker discovery a method for magnetic depletion of defective spermatozoa from bull semen, prior to extender addition and preparation of AI doses/straws has been developed. Commercially available nanoparticles coated with antibodies against ubiquitin, found exclusively on the surface of defective bull sperm is being employed. A second type of particle coated with lectin PNA that binds to acrosomal glycans exposed by premature sperm capacitation or acrosome reaction/damage is also in progress. Alone, or in combination, these nanoparticles were mixed with freshly collected semen for 15 min during the initial semen cooling, then removed by a strong magnet prior to semen extension and cryopreservation. This technique is simple and time efficient, has no requirement for equipment other than a magnet bar and uses commercially available components adding only cents to the cost of producing an AI dose. Evaluating the cytokine response during pregnancy following experimental challenge. 1. Investigation into the role of immunologic mediators during pregnancy following infection with the abortigenic bacteria that causes Epizootic Bovine Abortion may ultimately lead to a better understanding of the role of cytokines in the maintenance and termination of pregnancy in cattle infected with a bacterial pathogen. The leukocytes and inflammatory mediators involved in termination of pregnancy are important considerations in the development of therapies aimed at maintaining healthy pregnancies in cattle during pathogen infection. 2. Research into the development of a safe vaccine, one that is non-pathogenic to the host and can be used in pregnant animals, will be aided by the results of this research. Understanding which immunologic mediators play a role in a protective immune response against this pathogen will help tailor research into the development of a possible recombinant vaccine that could serve this purpose. Interactions of the embryo, uterus and corpus luteum for sustenance of embryos. 1. IFNT protects early pregnancy in ruminants through paracrine action on the endometrium and also through endocrine action on the CL and peripheral blood cells through enhancing luteal resistance to PGF. 2. A highly specific radioimmunoassay has been developed for IFNT, which has different levels of detection in uterine flushing compared to serum. IFNT leaves the uterus and has endocrine action on extrauterine tissues such as the CL and peripheral blood cells. 3. The endocrine mechanism of action of IFNT might entail up regulation of ISGs as well as cell survival genes to assist with providing resistance of the CL to PGF, but also in longer-term resilience in context of continued production of progesterone. 4. IFNT may also induce luteal resistance to PGF through induction of ISGs that prevent downregulation of genes encoding LHCGR and PTX3. Stabilization of LHCGR would support steroidogenesis and continued production of P4, whereas stabilization of PTX3 may prevent cell death and mediate inflammatory responses in a humoral rather than cell-mediated mechanism. 5. The direct assessment of gene function within the placenta has not been feasible in non-rodent species, especially in ruminants. A good case in point is that the placental lactogen has investigated for nearly 50 years, yet a direct function or relative importance has never been established. This is true for a plethora of genes expressed by the placenta. We know have the tools in hand to create transgenic placenta in sheep, that should allow the direct assessment of the impact of placental lactogen deficiency on sheep fetal growth near term (135 dGA). This technology can also be applied to other genes expressed within the ruminant placenta and/or fetus. A Bovine Model for Polycystic Ovary Syndrome. 1. A sub-population of cows within the UNL physiology research herd have androstenedione (A4) excess within follicular fluid. These cows appear to leave the herd at a 2:1 ratio compared to cows that have low A4 due to not having a calf. We are further characterizing these cows to determine if there is altered gonadotropin secretion, theca and granulosa cell gene expression or abnormal negative feedback of steroid hormones. We are also trying to determine if administration of CIDRs or progestins affect their profile of steroids in follicular fluid and how oocyte development may also be affected. Granulosa cells activate different signal transduction pathways dependent on follicle health status and ability to convert androstenedione to estrogen resulting in different steroidogenic profiles for beef cattle follicles. 1. We are investigating cows that have been treated with different progesterone or progestin as well as those with different concentrations of androgen in follicular fluid to understand how signal transduction pathways in granulosa cells may be altered. Prostaglandin F2a Activates Stress Response Signaling and Induces Expression of Activating Transcription Factor 3 (ATF3) in Bovine Large Luteal Cells. 1. Activating transcription factor 3 (ATF3) is a stress-adaptive gene that regulates proliferation or apoptosis under stress conditions. It appears to be upregulated during CL lysis initiated by PGF2alpha. This gene appears to suppress transcription of genes however, it does not appear to affect steroidogenic enzymes but does suppress progesterone production thus, it may work through increased cholesterol metabolism or transport. Understanding the basic mechanisms involved in CL lysis may allow for alternative methods to be developed to regulate the bovine estrous cycle. Loss of Vascular Endothelial Growth Factor A (VEGFA) isoforms in Granulosa cells using pDMRT-1 cre reduces female fertility by arresting follicular development and reducing number of follicles ovulated and affecting litter size. 1. Novel mouse lines where VEGFA isoforms have been knocked out in granulosa or Sertoli cells have been generated to delineate the functions and necessity for VEGFA isoforms in ovarian function. The actions of these VEGFA isoforms appear to have dramatic impacts on female reproduction. Loss of all VEGFA isoforms in Granulosa and some germ cells causes reduced fertility with smaller litter sizes and more days from mating to parturition. There are also less antral follicles developing with increases in mRNA abundance for FOXO3a, reduced estrogen concentrations in plasma, and smaller ovarian weight with a tendency for less CL. These all suggest that loss of all VEGFA isoforms causes subfertility by causing altered follicular development resulting in less follicles ovulating and smaller litters. Poor maternal nutrition affects postnatal growth and development of lambs. 1. Poor maternal nutrition due to over-feeding during gestation leads to increased size of lambs at birth, increased heart size and increased insulin which may predispose these offspring to metabolic disorders later in life. 2. Poor maternal nutrition due to under- and over-feeding during gestation impairs muscle development during postnatal growth. The reduced muscle growth may be due to altered gene expression within the muscle and satellite cell number and/or function. 3. Poor maternal nutrition during gestation alters the expression of key genes involved in the ability of mesenchymal stem cells to differentiate into adipocytes and osteoblasts. This could explain the differences in bone and adipose tissue development between offspring from control versus under- or over-fed mothers during gestation. 4. Poor maternal nutrition during gestation alters circulating concentrations of GH, IGF-I, insulin and leptin, which may contribute to, altered growth observed in offspring from these mothers. Characterization of the Pramel1 gene during spermatogenesis. The PRAME/PRAMEY gene family is essential for spermatogenesis and male fertility. Characterization of the bovine and ovine Y chromosome sequence, organization, and gene content provides not only important comparative mapping information for studying Y chromosome biology and evolution, but also male-specific genetic markers that will allow us to identify high and low fertility bulls in a sire selection program. Regulation and regulatory role of WNT signaling in potentiating FSH action during bovine dominant follicle selection. Investigation of the role of the Wnt signaling in regulation of follicular development may ultimately lead to development of new technologies to manipulate the follicular selection process in cattle and potentially enhance efficiency of superovulation. Expression profiling and potential functions of protein kinase C delta during early bovine embryo development 1. Recent work implicates PKC-delta as a mediator of embryonic development, interferon-tau expression and early placental development in cattle. Correlations Between PAG Concentrations, Pregnancy Loss and Milk Production in High Producing Holstein Cows 1. An initial assessment of a large database determined that placental and luteal insufficiencies are detected in dairy cattle prior to pregnancy failure. Continued analysis of this dataset will explore the impact of peri- and post-partum illnesses on pregnancy outcomes in dairy cattle. Impact of the number of estrous cycles before the start of breeding on reproductive performance of beef heifers and effects on subsequent re-breeding success. Attainment of puberty before start of breeding was associated with increased heifer pregnancy rate, with little or no benefit from having more than one estrous cycle prior to start of breeding. In contrast, second season pregnancy rate was positively associated with number of estrous cycles expressed prior to the start of a heifer's first breeding season. Circulating bovine pregnancy associated glycoproteins (bPAGs) are associated with late embryonic/fetal survival but not ovulatory follicle size in suckled beef cows No relationship exists between serum bPAGs and ovulatory follicle size or embryo stage/quality at ET; however, cows that lost an embryo after d 28 had lower concentrations of bPAGs on d 28 compared to cows that maintained pregnancy. Activation of the CXCL12/CXCR4 chemokine axis may drive vascularization of the ovine placenta The CXCL12/CXCR4 signaling pathway is activated during implantation and placentation in sheep and is likely playing a role in the communication between trophoblast cells and the maternal endometrium, specifically driving vascularization Membrane progesterone receptors are up regulated in caruncle tissue from obese compared to control ewes PGE1 and PGE2 functions in rats and sheep An essential fatty acid deficiency increased PSP and decreased the deciduomata response in PSP rats. Both PGE1 and PGE2 increased in pregnant ewes support PGE1 and PGE2 as antiluteolysins. PGE1 or PGE2 prevents progesterone-induced premature luteolysis by preventing a loss of luteal unoccupied and occupied LH receptors. Maternal obesity and overnutrition is associated with reduced systemic progesterone during the estrous cycles of adult female offspring and a marked increase in insulin resistance at midpregnancy. Mature ewes born to obese/overfed mothers (F1-MO ewes) exhibit marked greater insulin resistance with elevated blood concentrations of glucose and insulin throughout pregnancy when compared to daughters born to control fed mothers even when both groups were only fed to requirements throughout pregnancy. F2 generation born to these F1-MO ewes may exhibit the same metabolic alterations (hyperphagia, increased adiposity, and increased insulin resistance) as their mothers, resulting in a transgenerational programming effect. Dexamethasone (DM) administration in F0 ovine pregnancy eliminates the postnatal (PN) plasma leptin (L) peak in F2 offspring (OFF) and results in increased appetite and weight gain. Maternal obesity and over-nutrition alters the fetal sheep heart transcriptome We have previously demonstrated that maternal obesity and overnutrition increases fetal left ventricular heart weight and reduces cardiac work load capacity. Transcriptome profiling revealed differential expression of hundreds of genes some of which are known to be relevant to sodium, potassium and calcium transport suggesting that maternal obesity may influence sheep cardiac development and function. Role of Early Postnatal Leptin Surge in Setting Appetitic Centers There is strong evidence that the satiety hormone leptin, produced predominantly by adipocytes has effects on the maturation of the hypothalamus circuits controlling food intake. When these offspring reached adulthood, and were subjected to ad libitum feeding, they exhibited increased appetites, adiposity and insulin resistance. Obesity in pregnancy: implications for the next generation Late gestational low dose dexamethasone treatment programs arterial hypertension and vascular dysfunction in adult female sheep offspring Administration of exogenous glucocorticoids to pregnant ewes on days 103/104 of gestation programs arterial hypertension in adult female offspring. Late gestation fetuses of obese/overfed ewes exhibit increased adiposity, and elevated serum concentrations of triglycerides and cholesterol, as well as an increased PEPCK expression and reduced glycogen in their livers when compared to similar age fetuses from ewes fed only to requirements. Reduction in maternal feed intake only to requirements from day 28 through the rest of gestation in obese/overfed ewes returned all of these values to those of controls by late gestation, suggesting a beneficial effect of diet reduction. Female-oriented rams exhibit more neural activity along the olfactory pathway when exposed to sexually evocative stimuli than do sexually inactive rams. Exposure to sexually evocative and neutral odors elicited differential activity only in cortical amygdala in female-oriented rams. Activity in the cortical amygdala may alert rams and may account for the different behavioral response when sexually active rams are exposed to either estrous or non-estrous females. The lower neural activity in sexually inactive rams suggests inactive rams do not cue to sexual stimuli as sexually active rams do. The differential neural activity may account for the lack of behavior in the presence of estrous ewes. The presence of appetite hormones in breast milk may be important in infant appetite regulation. Additionally, changes in concentration of GLP-1 and Ghrelin during a single feeding may signal satiety in the nursing infant. Fetal adrenal demedullation at 0.7 gestation alters fetal growth and metabolism at 0.9 gestation in an ovine model of placental insufficiency and intrauterine growth restriction. Norepinephrine is required for normal ß-cell development and function. Chronic excess of NE as found in IUGR fetuses potentiates the actions of hypoglycemia and hypoxemia to suppress insulin secretion and fetal growth. A heterobivalent ligand composes of GLP-1 and adrenergic antagonist, yohimbine, exhibits high specificity for ²-cells in vivo. Multivalent ligands can be expanded and used to develop other ²-cell specific targeting agents or reproductive specific targeting agents. Temperament measured using exit velocity or pen score is heritable (0.27 to 0.49). This is adequate for selection to improve temperament of cattle. If only one method is to be used pen score has the greatest heritability (0.49). Calves can be effectively pen scored as single animals or in groups up to 5 animals. The correlation between single animal and group pen scores was 0.79. Prenatal transportation stress applied on days 60, 80, 100, 120 and 140 of gestation resulted in: a. More temperamental calves on days 14 and 28 after birth and through weaning. b. Only minor differences in complete blood cell counts due to prenatal stress but significantly (<0.01) greater numbers of basophils and a tendency (P=0.10) for greater numbers of neutrophils in females compared with males. c. Response to vaccination in white blood cell counts was reduced in prenatally stressed heifers but not in bulls. d. Prenatal stress and temperament alter metabolic response to an endotoxin challenge in Brahman bull calves. Circulating glucose was greater in prenatally stressed calves and in temperamental calves. Following endotoxin challenge insulin was lower in prenatally stressed calves. Cows that received controlled energy (CE) diets during the last 3 wk prepartum had shorter days to pregnancy (DTP) than cows that consumed high energy (HE) diets in this time period, which may be attributable in part to increased NELI in the first 4 wk postpartum for cows that received CE diets in the close-up (CU) period. In addition, lower BCS loss in the first 6 wk and slightly greater glucose concentrations at wk 3 may have contributed to improved reproductive performance. Energy- limited cows had lower liver TG concentrations at wk 2, which led to fewer DTP. A strategy of CE prepartum may have a favorable impact on both health and reproductive performance. Research evaluating the impact of CE prepartum on more specific reproduction variables, such as progesterone concentrations, ovarian function, time to first ovulation, and embryonic death is needed. Objective 2: Development and application of methodologies to improve animal health, well-being, and reproductive efficiency of domestic ruminants in order to allow sustainable operation of production systems in the western United States. Administration of FSH increased the number of follicles aspirated, oocytes collected, proportion of grade 1 to 3 oocytes, and number of grade 1 to 3 oocytes per cow. Administration of FSH, however, did not affect the ability of the oocyte to cleave or develop into a blastocyst or affect the characteristics of subsequently developed blastocysts. Reducing progesterone concentrations during follicular development increased the number of follicles present at aspiration but did not impact quality of oocytes recovered or the ability of the oocytes recovered to cleave and develop into a blastocyst. Of interest, blastocysts generated from oocytes collected in cows with low progesterone concentrations were advanced in development had greater number of blastomeres following in vitro embryo production. In beef heifers, reducing progesterone concentrations within an ultrasound-guided ovum pick-up protocol that included FSH administration resulted in more oocytes collected and increased the number of grade 1 to 3 oocytes recovered per female. It was demonstrated that a majority of Bos indicus (Nelore) heifers will conceive at one year of age and rebreed effectively as primiparous cows with sufficient nutritional management and hormonal intervention.