Brief Summary of Minutes of Annual Meeting

8:00 am Chair Tracy Davis called the meeting to order. Per the renewal of our project, the official number of our meeting is W2112. Secretary is Kimberly Vonnahme, and Member at Large is Ryan Ashley. Members introduced themselves. Special welcome to our new members: Allen Bridges, Kristin Govoni, Nathan Long, Bret Hess (administrator) A big congratulations to Dr. Ron Randel for being the 2012 recipient of the L.E. Casida Award at the National ASAS meeting. 8:15 am There was a phone conference call with Mark Mirando. Briefly, Dr. Ramaswamy was appointed as the Director of NIFA on May 7, 2012. The US budget appropriations for AFRI funding, Higher Education Grants and Smith-Lever grant funding was discussed. Mandatory and endowment funding are subject to reauthorization within the Farm Bill. For competitive programs, Foundational programs will have FY 2012-2013 combined and RFA is expected to be released August 15, 2012. The reported decline in the last funding cycles success rate was due to the decrease in the budget as well as the increase in total allowable budget (ie. $350K to $500K in budget ceiling for the grants). There will be an increase in the allowable indirect costs from 22% to 30%. The USDA-NIH Dual purpose grant proposals have been reviewed and funding decisions should be made in the near future. September 20, 2012 is the new application deadline. Mark will forward the email address for members to encourage that the NIH-USDA Dual Purpose continues if that is our desire. There was a stakeholder workshop in Baltimore (March 2011) as well as a ARFI Stakeholder Listening Session in DC on Feb 22, 2012. Questions from the floor: Foundational grants: Will the cap still be $500K? Answer: Not sure what will be in the RFA. With a $500K cap, direct costs will be $40,000 less with the 30% IDC. This increase in IDC basically results in one less grant being funded. Ecology and infectious diseases: Is this a new program? Answer: NIFA just started to participate in this program. 8:30 am Mike Teglas moved to approve the minutes; Brenda Alexander seconded the motion. Motion passed unanimously. Brenda Alexander nominated Mike Teglas for a new member at large. Members voted unanimously in favor, and Mike is our new member at large. 8:35 am New Business: Each station gave updates on what is new at their home institutions/departments. NMSU: There is the potential for a new reproduction position in the future. Milt Thomas has accepted a position at Colorado State University. Texas A&M: Possible new reproduction position may be open in the future. Tom Spencer has accepted a position at Washington State University. Utah State: College of Ag has built a new facility that houses animal, veterinary and dairy sciences, it is proposed that a new USDA-ARS facility will be built nearby. A new vet med collaboration between Utah State and Washington State has been established with students starting their studies at Utah State and finish at WSU. USDA Miles City: Mike McNeil has retired and there may be an advertisement to refill his position. University of MN: Jon Wheaton is retiring. Allen Bridges is our new member from UM. There is the possibility that a new reproduction position will be advertised in the future. There is a possibility of 2 100% extension positions in the future (reproduction at Grand Rapids and feedlot at UM). University of Wyoming: Governor has announced that there will be an 8% budget cut for the University of Wyoming. Min Du and Meijun Zhu from the Department of Animal Science have left to accept positions at Washington State University. There is hope that these positions will be filled in the future. Bill Murdoch has announced that he will retire next summer. Bob Stobbart will also be retiring. University of Alaska: The School of Natural Resources has been placed on financial probation, and the Dean and Director is retiring. Extension is in good shape. Ag economics position will be coming open. University of NV-Reno: Extension budget was cut in half. Currently, there is an animal nutritionist position (emphasis in range) open in the Department. There are plans to advertise a nutritional-epidemiologist sometime in the near future. UCONN: Department Head of Animal Sciences, Dan Fletcher, has retired and Steve Zinn is the interim department head. Three new positions have been hired-- Functional genomics, equine position, and new lecturer (100% teaching). There is a new president at the university who has the intention of hiring 250 new faculty positions over the next 4 years. It is hoped that Animal Science will secure some of those positions. University of AZ: The Department Head has accepted the position as the Ag Experiment Station Director. There is some discussion to combine the Animal Sciences Department with Veterinary and Microbiological Sciences. North Dakota State University: Dr. Gerald Stokka has been hired in a livestock stewardship position. A new Beef Cattle Research Center is now open. Dave Buchanan has accepted the position as the Associate Dean of Academic Affairs within the College of Ag. University of Idaho: There are 2 Beef extension positions and a veterinarian position open. Town of Moscow will be rezoned which may run through the dairy facility. There is a potential for new facilities. Colorado State: There is a new department head, Kevin Pond, at Animal Sciences; Milt Thomas, chair in animal genetics and member of ARBL: ARBLcrosses over to Animal Sciences. The position for the Alexander endowed chair will open. The equine reproduction lab burned. Redesigning the equine building is underway. University of Nebraska: There is an expectation that the student enrollment needs to get to 30,000. In order to support that many students, there will need to be an increase in resources for classes, namely lab classes. UNL is now in the process of searching for extension dean. Archie Clutter is the new Research Dean. New vice chancellor is Ronnie Green. Animal Science just had a departmental review and it is recommended that a stem cell/epidemiologist position and a stress physiology position be added. 9:15 Comments from Bret Hess, Administrative Advisor Midterm review (this is in response in the review of the renewal of the project) 1) Be sure to demonstrate collaboration within the group. 2) Accomplishments, make sure we follow our listed objectives; remember to meet our milestones. This includes completing the estrous synchronization handbook. This should be completed by the midterm review. 3) Annual report: distill the highlights. Report our outcomes (i.e. a change in knowledge, skills and abilities) and impacts (i.e. how this impacts society, fill in knowledge gaps). 4) Reminder that the increase in indirect costs (IDC) are now 30%, which means that the modified indirect rate is now 42.86% to direct costs. Experiment station directors: Currently Bret Hess is the only animal scientist. Soon, there will be 2 additional (AK and AZ). We voted to have Milan Shipka serve as the W2112 administrative advisor. We have 60 days to get the report done and submitted. 9:30: nominations for the 2013 meeting: Fargo Bozemanwith the WSASAS Anchorage Overton, Texas 9:40 Station Reports (NMSU, UW, CSU, USDA-Miles City, U of MN, Texas, U of AK, Utah State, UNR, see below) 5:00 pm Adjourn for the day Friday, June 1, 2012 7:00 am Breakfast in Valley View 1734 8:00 am Tracy Davis called the meeting to order. Bret Hess will distribute a request for further reports from members that did not submit a report. These will be due on June 15th. Continued station reports (UNL, UCONN, Univ of AZ, NDSU, Univ of Idaho; see below) 11:00 am Discussion of collaborative plans for 2012-2113 Nebraska & Washington: Dept of Health & Human Services grant 2009-2011. Joint publication has been submitted and submission of NIH RO1 grant June 15 titled Role of VEGFA Isoforms on Spermatogonial Stem Cell Renewal. Idaho & CSU: Evaluate enhanced luteal function on conceptus length and IFNT production in lactating dairy cows. Idaho & Colorado: Use of next generation of sequencing using isolated gonadotropes. Idaho & Colorado: Adenoviral delivery of dominant negative dynamin into the ovine pituitary and the effects of estradiol release on LH release. Colorado & Idaho: RIA for IFNt and Colorado providing reagents for RIA to Idaho. Wyoming, Colorado, Hawaii, & Texas: Continue to analyze the data from the PGE project and write manuscripts. New Mexico & Arizona: Develop an assay to evaluate progesterone receptors in mitochondria. Colorado & Wyoming Project: Develop a more efficient and cost effective method to synchronize estrus in cattle. Arizona & New Mexico: Share data about stress response and impact on placental factors and implantation. Arizona & Florida: Share data from microarray of bovine blastocysts and trophoblast cell line. Arizona & Wyoming: Insulin secretion in isolated pancreatic islet cells. Arizona & Colorado: Evaluate vascularization of pancreatic islets. New Mexico & Florida: Share bovine trophoblast cell line and compare genomic data obtained from bovine placenta and endometrium. Montana (Miles City), Michigan, & Missouri: Investigate expression of cathepsin, inhibin, and follistatin expression in bovine follicles related to the role of follicle size at the time of ovulation on establishment and maintenance of pregnancy. Montana (Miles City), South Dakota, Missouri, & Washington: Investigate the role of estradiol on pregnancy establishment and uterine expression in ovariectomized cows and intact heifers. Florida & Nebraska: Examine VEGF in bovine endometrium and placenta. Florida & Washington: Effect of inhibiting FGF in reproductive tract during early pregnancy in cattle. New Mexico & Montana: Metabolic hormone patterns in ewe lambs with differing RFIs. New Mexico & Colorado: Nutrient restriction vs. metabolic hormone patterns. New Mexico & Colorado: Thyroid involvement in BVD infected cows. New Mexico & Arkansas: Metabolic hormone profiles in cows consuming endophyte infected fescue. New Mexico & Ohio: Follicular fluid IGF. Wyoming & Montana: Meta-analysis of birth type on ewe reproductive performance. Colorado & Ohio: ISG mRNAs in maternal blood and IFNt in conceptus. Colorado & Washington: Use of mouse ISG15 knockout model to understand the role of ISG15 in bovine uterus. Michigan & Florida: Extended oocyte/embryo culture. Ohio & Pennsylvania: Role of MX and IFNt in cattle. Arkansas & New Mexico: Pasture systems and reproduction. UDSA-Miles City, Minnesota & Idaho: Effects of desoerlin on reproductive efficiency in beef cattle CSU & NMSU: Maternal T3 and T4 in cows carrying BVDV infected or control fetuses. CSU and Texas A&M: Final validation of a IFNT RIA NMSU & NDSU: Endocrine profiles during pregnancy and lactation in ewes Texas & USDA-Miles City: Examine longitudinal data for heifer and feed efficiency on long term reproductive and productive characteristics. Utah & CSU: Sharing and analyzing microarray data from CL from pregnant and nonpregnnat sheep Utah and UW: DUD and high performing ram data analyses USDA-Miles City & Washington: identification of fertility markers in beef cattle USDA-Miles City, Ohio, Idaho, & MN: Role of progesterone in determining oocyte competence and use of GnRH agonists in manipulating progesterone to improve fertility in beef and dairy cows. Ohio, USDA-Miles City, Idaho, & MN: Estrous synchronization projects and re-writing of Estrous Synchronization Handbook in cattle NMSU & USDA-Miles City: Progesterone receptor in the uterus NE & MN: Link between obesity-oocyte quality UCONN & NE: Collaboration on an obesity model and bone development UCONN & NDSU: somatotropic axis and fetal development WY & AK: Look at cortisol in reindeer NDSU & USDA-Miles City: Doppler blood flow in cattle It was moved and seconded that the 2013 meeting of W2112 be at Miles City (USDA-ARS). Majority voted to have meeting at Miles City.

Objective 1: Examine molecular, metabolic, genomic, endocrine, and immunologic mechanisms influencing testicular and ovarian function, reproductive behavior, conception rate, embryo and fetal development, attainment of puberty, and effects of climate/season on reproductive patterns of domestic ruminants. 1. During early gestation in ewes, CXCL12, CXCR4, and select angiogenic factors and their receptors are elevated in fetal extraembryonic membranes and maternal caruncles, underscoring the possible involvement of this chemokine system in placentation. 2. Numerous tissue-specific alterations in ovine MPR expression occur during early pregnancy, which correlates with rising P4 levels and when fetal-maternal placental interactions are established. 3. IFNT protects early pregnancy in ruminants through paracrine action on the endometrium and also through endocrine action on the CL and peripheral blood cells in context of enhancing luteal resistance to PGF and the maternal ability to combat viral infections, respectively. 4. ISG15, Bcl-XL, Akt, and XIAP mRNAs are up regulated within 24 h in response to roIFNT treatment of isolated small, large and mixed luteal cells, and this induction is independent of the actions of PGE2, OXT and PGF. These results are similar to the results obtained in previous in vivo studies using osmotic infusion of roIFNT into the uterine vein of ewes. Endocrine 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. 5. Knockdown of SLC7A1 attenuated conceptus growth and development, whereas knockdown of NOS3 and ODC1 only partially suppresses conceptus development. This is likely due to compensatory effects of NOS1/NOS2, other cationic amino acid transporters and production of putrescine via the arginine decarboxylase and agmatinase biochemical pathway to support conceptus development. 6. TLR7 and TLR8 knock down conceptuses were developmentally retarded. Knock down of TLR7 and TLR8 also impairs release of IFNT in uterine flushings (P<0.01). 7. 5. Transmission electron microscopy revealed degenerative and necrotic changes in murine anti-mesometrial decidual cells and trophoblast cells at 7.5 dpc; however the decidualization responses and embryos appeared normal at this time. ISG15 is expressed in multiple species such as bovine, ovine, human and mouse that have divergent forms of implantation. ISG15 functions in vascular development; and immunomodulation. The high incidence of embryo mortality in Isg15-/- mice suggests that ISG15 plays an important role in uterus:conceptus communication during implantation, which may also extend to function in ruminants. 8. Persistent infection with BVDV impairs tissue level but not cell level bone formation. Abnormal trabecular modeling is secondary to reduced numbers of osteoclasts. The factors responsible for the periodicity of the transverse bands noted in PI femur are unknown but may be related to osteoclast generation times. 9. As a nuclear protein, we initially hypothesized that PRR15 might act as a co-regulator of transcription. However, our pull-down data coupled with recent localization studies identifying PRR15 in nuclear bodies suggest its mechanism of action is at the post-transcriptional level. Using shRNA infection of trophoblast cells we determined that diminished PRR15 expression impacted a number of pathways, consequently impacting trophoblast proliferation and apoptosis. Our recent results, as reported here, help explain the lethal phenotype we observed in sheep conceptuses infected with an shRNA that targeted PRR15 expression in the elongating sheep conceptus. 10. The pluripotency factor Lin28 is expressed in the early sheep conceptus, and the relative expression pattern of Lin28A and Lin28B differ, suggesting that these two RNA binding proteins may have distinct functions in the developing placenta. Initial attempts to diminish the mRNA concentration of Lin28B in the sheep conceptus was not successful, but has led us to the design and testing of new targeting vectors which will enhance our ability to alter gene expression in the ruminant placenta. 11. We continue to examine the impact of maternal undernutrition during twin pregnancies on fetal development and postnatal growth. Our recent studies indicate that maternal Ghrelin is altered by undernutrition, as we reported for fetal Ghrelin concentrations last year. The elevated maternal Ghrelin was not diminished following realimentation at mid-gestation, and carried out into the postnatal period, suggesting long-term impacts on maternal metabolism and appetite. Since multi-fetation is the norm in production flocks, examining the impact of nutritional imbalances during pregnancy will provide better insight to the impact maternal/fetal programming in a production setting. 12. Temperamental calves have greater stress hormone concentrations at or near weaning than calmer herdmates. Temperament did not affect concentrations of immunoglobulins but sex of calf does with steers having higher concentrations than intact bulls or heifers. Gender has an influence on circulating concentrations of stress hormones with heifers being greater than bulls. 13. Temperament at maturity in cows can affect birth weights of their male calves. Temperamental cows producing bull calves had lighter birth weight calves than did calmer herdmates. 14. Selection of replacement females for residual feed intake or residual average daily gain does not affect birth weights of their calves. 15. Longer duration transport (24 hours) increased shrink in body weight more than shorter (5.5 hours) transport but feeding behavior and performance were not different. 16. Calves mobilize intramuscular fat during transportation and the quantity of intramuscular fat was diminished for up to 28 days after transportation. 17. A statistical framework was developed to use gene expression data to identify Gene Ontology biological process terms with significantly different activity profiles across multiple embryonic developmental stages. While motivated by an RNA-Seq experiment, the statistical framework can be applied to any gene expression platform. 18. The most important factors affecting fertility to GnRH induced timed AI were elevated serum estradiol concentration at the time of AI, elevated serum concentration of progesterone on d 7, elevated serum concentration of progesterone on d -2, and ovulatory follicle size. Fertilization failure in cows receiving GnRH induced ovulation of single follicles was 10%. Young cows have greater fertilization success, but are less likely to maintain a pregnancy than older cows. Evaluation of serum estradiol concentration in a single blood sample collected at TAI is more valuable as an indicator of a cows fertility than measure of serum concentration of progesterone in two blood samples collected before synchronization. Age of the ovulatory follicle (between 8 to 13 d old) did not affect pregnancy rates. 19. Cows that received GnRH induced timed AI without an increase in serum estradiol concentration develop embryos that are less capable of establishing a pregnancy (preventing luteolysis around d 17 based on IFN stimulated gene expression) and incapable of maintaining pregnancy to d 30 of gestation. With synchronization programs that use estrous detection and AI for a period before timed AI of remaining heifers my result in improved pregnancy rates among timed AI cows by adding an estradiol treatment at timed AI. 20. Body condition score of superstimulated embryo donor cows impacted various aspects of embryos recovered. Embryos recovered from cows with a body condition score of 6 tended to be advanced in stage of embryo development and total number of grade 1 and 2 embryos collected per cow also tended to be increased. In addition, embryo donors with a body condition score of 6 yielded increased numbers of transferable embryos per cow than did embryo donors with a body condition score of 4. 21. Although increasing body condition score of the embryo recipient increased the diameter of the ovulatory follicle induced to ovulate prior to embryo transfer and resulted in a larger CL that produced greater concentrations of progesterone, such follicular and endocrine differences were not reflected in different pregnancy rates between embryo recipients in either a body condition score 4 or 6. Moreover, body condition score of the embryo donor did not impact probability of pregnancy establishment. 22. Feeding DG at 1.2% of BW per day to first-parity heifers during the third trimester of pregnancy appeared to elicit some degree of developmental programming to female offspring. Heifer calves from pregnant females receiving high amounts of distillers grains were heavier at weaning, had greater skeletal growth and were heavier at onset of puberty. In addition, AI pregnancy rates were increased, but limited numbers of heifers in this study limits conclusions that can be made from this categorical variable. 23. Understanding these mechanisms and ultimately their potential impact on blastocyst survival, fetal development and calf growth are important to breeding management of reindeer, especially if techniques like artificial insemination are to be applied. 24. The impact of the timing of breeding on gestation length, pregnany rate, calf sex, calf birth weight, and calf survival all have important implications on the successful management of muskoxen in captivity. 25. The protective effect against subsequent infections in cattle following an abortion due to EBA appears to last up to 2 years following an abortion, then abruptly stops without continued exposure to the pathogen. 26. Infection with the agent of EBA stimulates a mild hematologic response characterized by leukocytopenia, lymphocytopenia and monocytosis early (week 1) and late (week 14) in infection but leukocyte cell numbers do not rise or fall beyond what are considered normal parameters for pregnant cattle. 27. The bacterial pathogen that causes EBA cannot be detected by molecular methods in the circulating blood of infected heifers during the entire course of infection. 28. We have made significant strides in understanding the specific implications of maternal obesity on offspring health, growth efficiency, glucose-insulin dynamics and reproductive function. Results from these studies shed light on the specific physiological mechanisms involved, as well as lead us to management practices to optimize offspring health and production efficiency. These studies provide strong evidence that the optimization of maternal nutrition and the associated uterine environment will contribute to the birth of healthy, growth-efficient offspring, which is vital to livestock production efficiency. 29. The expression of male reproductive behavior is essential for the continuation of most species, but is of particular importance in the sheep industry where the use of artificial insemination is limited. A robust sexual response to the presence of an ovulating female is the first step to species survival. Among males, expression of sexual behavior is a continuum that includes males that lack sexual interest and those that show exuberant sexual interest in other males. We have reported evidence that in multi-sire flocks rams siring higher than expected numbers of lambs made up 23  29% of the ram population but sired from 39.4 to 70% of the lamb crop. Low performing rams (23  43% of the ram population) sired from 3.2 to 16.2% of the lamb crop. We are using cellular techniques which will help identify neural processes which differ among male oriented and female oriented rams which express either high or low levels of reproductive behavior. 30. Female oriented rams appear to process sexually evocative olfactory stimuli differently than rams lacking sexual interest which may help explain the absence of sexual behavior in rams categorized as asexual. 31. The differential expression of dopamine synthesizing enzymes in sexually active and non-active rams may reflect the dichotomous role of dopamine in the expression of sexual behavior. 32. Genome-wide association studies (GWAS) may benefit sheep producers in the calculation of molecular breeding values (MBV). Initial analyses of many production traits revealed significant SNPs on OAR4 with average daily gain, staple length, wool grade, and fleece weight. Refinement of these associations is now in process. 33. Increased glucocorticoids during gestation reduced HPA axis activity, but increased baseline activity in female offspring and this effect is carried over into a subsequent generation. 34. Maternal obesity increased expression of adipogenic factors (PPAR ³ and AP2) and 11²SDH1 (an enzyme that controls local availability of cortisol) in perirenal, and subcutaneous adipose tissue. There were depot specific effects for LPL, a marker of adipose maturity. Angiogenic factors were decreased in obese vs control fetuses in perirenal and increased in subcutaneous adipose. 35. Maternal obesity-induced alterations in maternal and fetal glucose-insulin dynamics and fetal adiposity can be alleviated by early maternal dietary intervention. 36. Differential expression of genes relevant to sodium, potassium and calcium transport may underlie functional changes in fetal hearts caused by maternal obesity. 37. Maternal obesity leads to increased appetite and adiposity that is associated with increased adipocyte diameters and increased rate of fatty acid synthesis in overnourished adult male offspring. 38. Increased adiposity of adult male offspring from obese mothers is associated with decreased leptin receptor expression by GH secreting pituitary cells in association with increased expression of pituitary PPAR-³, leading to decreased GH secretion and reduced expression of liver IGF-1. 39. Although prolonged decreases in maternal uteroplacental blood flow may contribute to maternal caused fetal growth restriction at midgestation in nutrient-restricted ewes, large conductance Ca+ dependent K+ channels (BKCa) expression in the uterine artery during the first half of pregnancy was similar in nutrient restricted and control ewes. Further studies of maternal nutrient restriction and uteroplacental blood flow are warranted. 40. Novel mouse lines where either VEGFA isoforms or the co-receptor to VEGFA angiogenic isoforms (Nrp-1) have been knocked out in granulosa or Sertoli cells have been generated to delineate the functions and necessity for appropriate balance of VEGFA isoforms in ovarian and testis development. The actions of these VEGFA isoforms appear to have dramatic impact on male and female reproduction if they are not in appropriate balance. 41. Loss of all VEGFA isoforms in Sertoli and some germ cells causes reduced fertility with smaller litter sizes and more days from mating to parturition and a greater amount of time from 1st parturition to 2nd parturition in heterozygotes. 42. Loss of VEGFA isoforms in granulosa cells causes reduced fertility with decreased litter size in heterozygote females and longer days from mating to 1st parturition and 1st parturition to 2nd even in heterozygote females. 43. Sertoli cells and deactivation of VEGFA angiogenic isoforms appears to have little effect on organ weights; however morphology of the testis is altered and genes critical for spermatogonial stem cell niche maintenance are dramatically reduced suggesting that too much anti-angiogenic isoform actions are detrimental to reproductive lifespan and male fertility. 44. Injection of VEGFA isoforms prior to spermatogonial stem cell niche establishment in a transient manner affects genes that are critical to apoptosis and spermatogonial stem cell homeostasis in the rat. Suggesting that altered VEGFA isoforms during development of the spermatogonial stem cell niche can influence early spermatogenesis. 45. Estrogen active follicles tend to have a greater ratio of VEGFA164 to VEGFA164b and a greater efficiency of conversion of A4to E2; while estrogen inactive follicle have greater mRNA abundance of VEGFA164b and are less efficient at converting A4 to E2. Furthermore, there is a positive correlation between the antiangiogenic VEGFA isoform and BCL2 and AMH which play a role in regulating follicle development. Thus, the balance of VEGFA anti and proangiogenic isoforms may influence follicular development and atresia. 46. There appears to be a relationship between AFC and BW through the first breeding season and progeny calf BW. The low birth BW in heifers with low AFC and in their progeny continues to support a possible link between genes that influence growth and development and establishment of the ovarian reserve. Thus, cows with greater AFCs may also produce offspring with greater birth weights and growth traits. Selection for AFCs as well as other traits reproductive traits early may aid producers in determining females to remain in the breeding herd. 47. Male fertility is critical to many animal production systems. We have demonstrated that VEGFA isoforms are important in early testis development to establish the spermatogonial stem cell niche alterations in the balance of these isoforms do affect male fertility. Thus, manipulation of these VEGFA isoforms may be economically important to increase reproductive efficiency in males for producers. 48. Female fertility is equally important in animal production systems. In appropriate balance of VEGFA isoforms also appears to impair female fertility through altered follicle development and potentially numbers of follicles available on the ovary to ovulate. Thus, manipulation of VEGFA isoforms or being able to understand how we may use them as markers may dramatically impact producers and their ability to either increase reproductive efficiency or select for more fertile females. 49. The VEGFA164b isoform in cattle appears to be a marker for follicle status, and health. Furthermore, it is correlated with genes that regulate follicle development such as AMH and BCL2. 50. Poor maternal nutrition due to under-feeding during the last half of gestation negatively impacts fetal growth and development as demonstrated by reduced body weight and reduced circulating IGF-I and IGFBP-3. 51. Poor maternal nutrition due to under- and over-feeding may negatively impact metabolic status of offspring as early as birth, but altering leptin production and serum lipid concentrations. 52. We continue to determine methods or therapeutics to alleviate the negative impacts of IUGR in livestock. a. Melatonin work: It is determined that melatonin supplementation can increase umbilical blood flow in nutrient restricted ewes. Melatonin treatment partially improved uteroplacental amino acid flux in nutrient restricted ewes to the level of CON-ADQ ewes, which may be mediated via increased placental blood perfusion and/or placental amino acid transporter density. Maternal melatonin supplementation and nutrient restriction interacted to effect fetal cardiomyocyte enlargement, whereby melatonin supplementation during maternal nutrient restriction may reverse left ventricular cardiomyocyte enlargement. However, this effect was absent in the right ventricle, which may potentially alter offspring cardiovascular function and performance. b. Realimentation in beef cattle: It appears that nutrient restricted impacts vascular reactivity at the level of the placentomes (i.e. caruncle and cotyledonary arteries). Current work on uterine blood flow measurements are under analyses, however, umbilical blood flow to day 85 of gestation is not impacted by maternal nutrient restriction from day 30 to 85 in beef cows. Protein supplementation: Supplementing protein during the last third of gestation in ewes appears to have positive impacts on feedlot performance of wether lambs, while having limited impacts on carcass quality. 53. Chronic administration of GnRH agonist beginning day 3 when the CL is developing through day 15 of the estrous cycle enhances size of the CL which is correlated with circulating progesterone concentrations. Furthermore, co-dominant follicles form during the first follicular wave and most ovulated and formed accessory CL. Finally, timing of the removal of the GnRH agonist does not alter size of the ovulatory follicle or length of the estrous cycle. 54. Outward symptoms of bacterial meningitis in sheep infected with either S. aureus or A. pyogenes are correlated with major hormonal shifts, especially associated with the time of estrus when progesterone concentrations decline, prostaglandin F2alpha and estradiol concentrations increase. 55. The results of studies conducted during the past year demonstrate that estradiol and selected phytoestrogens dramatically affect basal and GnRH-induced gonadotropin secretion from ovine pituitary cells in culture. Collectively these data suggest that phyto- and xeno-estrogens may influence the reproductive potential of domestic animals. 56. The results of studies conducted during the past year demonstrate that the PRAME/PRAMEY gene family is essential for spermatogenesis and male fertility. The gene family amplified in Eutheria and was transposed to the bovid Y chromosome during evolution, indicating that this gene family is even more important in male reproduction in bovid lineage. Characterization of the 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. Elimination of sires with genetic defects associated with reduced fertility at an earlier age will reduce overall maintenance costs for poor quality yearling and mature sires prior to breeding. 57. Investigation of the role of the CART peptide 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. 58. Investigation of the role of oocyte-derived follistatin in early embryogenesis support the utility of follistatin treatment as a potential tool to increase efficiency of in vitro embryo production and embryo quality and provide a viable platform to aid studies of the contribution of poor oocyte quality to infertility in cattle in a production setting. 59. EP4 receptors may be involved in preventing luteal regression to permit establishment of pregnancy. SIRT-1 (located in the nucleus) may be involved in regulating PGE synthesis by ovine caruncles. 60. The development of a hypothalamic structural atlas will support future studies by enabling others to identify specific centers in sections of interest, thereby reducing the total amount of antibodies and reagents needed to examine the immunocytochemical expression of specific neuropeptides of these centers. The ability to identify key structures such as the arcute nucleus based on histo-structure shows the utility of this atlas. The long-term goal of this research is to determine the physiological pathway whereby pheromones of ruminant males induce changes in the hypothalamic kisspeptin-GnRH system that accelerated the onset of the breeding season in sheep and other seasonally breeding mammals and resumption of ovulatory activity in postpartum, suckling cows. 61. During early pregnancy in the mouse, the corticosterone biosynthetic enzyme HSD11B1 is up-regulated in response to the implanting embryo. Implications are that the embryo establishes a glucocorticoid barrier around the embryo that may serve to: 1) modulate maternal immune function; 2) regulate nutrient availability; and 3) regulate trophoblast growth. Conditional deletion of the glucocorticoid receptor, Nr3c1, results in a subfertility phenotype that may derive from abnormal infiltration of immune cells. 62. Regeneration of the endometrium likely involves distinct mechanisms that include: 1) activation of a unique population of stem cells; and 2) mesenchymal-to-epithelial transition. 63. Tissues of the female reproductive tract abundantly express the E2A encoded product TCF3/E47. E2A is regulated by steroid (uterus) and gonadotropin (ovary) hormones in vivo. 64. Pgrmc1 plays an important role in female reproduction. Conditional deletion of this gene results in a subfertility phenotype stemming from disrupted folliculogenesis and formation of cystic structures in both the ovary and uterus that likely occur in response to aberrant cell cycle progression. 65. PGRMC1 is important for germ cell proliferation and completion of meiosis in males. The loss of PGRMC1 in proliferating germ cells results in a repetitive disruption of spermatogenesis due to altered cell cycling and accelerated germ cell differentiation. Pgrmc1 location on the X chromosome suggests it is a strong candidate for inherited male factor infertility. 66. Spermatogonial stem cells formed during neonatal life rapidly proliferate during the first weeks of life. The adult population of spermatogonial stem cells forms after puberty. 67. Testicular dysgenesis in alpaca and possibly other camelids is potentially associated with alteration of the formation of functional Leydig cells during puberty. High variance in the age of puberty in alpaca is associated with variation in testosterone production. 68. Prostaglandins are important regulators of conceptus elongation and mediators of endometrial responses to progesterone and IFNT in the ovine uterus. 69. Prostaglandins and IFNT from the conceptus coordinately regulate endometrial functions important for growth and development of the conceptus during the peri-implantation period of pregnancy. 70. IFNT and prostaglandins from the conceptus regulate endometrial HSD11B1 expression and activity that regenerates bioactive cortisol in the ovine uterus during early pregnancy to influence endometrial functions and conceptus elongation. 71. Feeding a high energy diet can advance some aspects of maturation in Holstein bulls. Technology to advance puberty and the age at which semen can be collected from bulls for use in AI has the potential to accelerate genetic improvement and take full advantage of genomic testing in AI sires. 72. In vitro methods to identify bull fertility would enhance reproductive efficiency and streamline methods for selection and identification of bull to use in AI. The use of heterospermic IVF was not an accurate method to predict competitiveness of different bulls in vivo when heterospermic insemination was performed. This finding suggests that alternatives to this approach to determine fertility of bulls for AI is necessary. 73. The lack of effect of follicle age on conception in beef cows and heifers provides crucial information to aid in development and refinement of estrous cycle control technologies. 74. Induction of puberty in Nelore heifers and reinitiation of estrous cycles in postpartum Nelore cows is essential to advance reproductive efficiency in this crucial breed to global beef production. Effective methods to achieve this end, and implement timed AI have the potential to substantially enhance the efficiency of beef production. 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. 1. Administering 5 or 10 mg of dexamethasone to ewes before breeding had no adverse effect on pregnancy rates and the 5-mg dosage may have actually been beneficial. 2. Use of a CIDR in combination with 500 IU PMSG effectively induced out-of-season cyclicity at 30 d postpartum in lactating anestrous Rambouillet ewes and approximately 37% of anestrous ewes bred to fertile rams lambed again in during the subsequent fall. 3. GnRH vaccination in elephants shows potential for contraception and management of uterine pathology in older elephants. 4. At least in elk, maternal antibodies to GnRH that are transferred to the offspring via colostrum do not have an impact on subsequent reproductive development of the offspring. Therefore, immunization against GnRH can be carried out irrespective of the physiological state of the female without concern for the normal development of her offspring. 5. Immunization against GnRH using GonaCon results in infertility up to three years following a single injection. In addition, immunized females show extended proreceptive behavior when exposed to males. 6. 12. Use of a conjugate prepared with a GnRH analog covalently linked to a recombinant pokeweed antiviral protein showed efficacy in reducing GnRH-induced LH release in mice and testis weight in mice. Such a compound may be effective for chemically sterilizing both males and females of a variety of species including domestic animals. 7. Feeding a yeast based supplement improves the acute phase response to an endotoxin challenge and increases availability of glucose which is required for the animal to respond to an endotoxin challenge. It is possible to feed a supplement before cattle are subjected to a stressor and improve their ability to cope with the stressor. 8. Spermatozoa characteristics of the ejaculate used to breed beef cattle may account for a significant percentage of pregnancy failures. 9. Uterine capacity for pregnancy was the primary limitation of fertility in a group of heifers identified with differential levels of fertility. Genomic wide analyses of difference between these heifers have revealed 7 markers associated with fertility in these heifers. 10. The 5-day CO-Synch + CIDR, PGF 6-day CIDR, and 14-day CIDR-PG protocols were all equally effective methods to synchronize ovulation in virgin beef heifers. Pubertal status at ovulation synchronization had the greatest impact on pregnancy success. 11. The aoEBA can be raised in vivo in SCID mice and infected cells produced in this manner can recreate the disease with clinical signs identical to those from infections using bacteria obtained from EBA aborted fetuses. 12. Infected cells from a murine source can be used as a whole bacteria vaccine with protective effects if injected into cattle prior to breeding. A dose dependent association appears to be correlated with early fetal loss in cattle previously sensitized to the aoEBA. 13. Increasing DDGS in the diet of rams did not have a negative impact on feedlot performance or carcass characteristics; however, in this preliminary study, sperm production may have been negatively affected, necessitating the need for additional research on the impact of distillers grains on ram development. 14. Development of technology to decrease injections and improve compliance for ovulation synchronization in dairy cattle has the potential to improve animal well being and increase reproductive efficiency. Intrauterine administration of GnRH is one potential method to achieve this end.