Brief Summary of Minutes of Annual Meeting

Chair Andrea Cupp called the meeting to order at 8 a.m. on Monday June 4, 2007. Participants were introduced and the minutes from the 2006 meeting were approved without discussion. Tod Hansen welcomed everyone to the Animal Reproduction and Biotechnology Lab (ARBL) and the Equine Reproduction Lab (ERL) at CSU. Tod provided historical comments and a description of ARBL and the ERL, the CSU Foothills Campus and a description of research and training program. The group thanked Tod, Terry Nett and the rest of the CSU group for local arrangements. Registration fee for the meeting was $50 per attendee. Colin Kaltenbach reported that we are completing year 1 of the W1112 multistate project and the termination date is September 30, 2011 and encouraged participants to submit their reports in a timely manner. All participants need to keep the email list up-to-date by making an email address changes on the NIMS system. The group asked Colin for an update on efforts to transition USDA Hatch or Experiment Station funds from the current system into a competitive funding system. Deb Hamernik will cover this in her comments. Colin reported that there is no effort or plan to stop multi-state projects or committees because these have a high priority. Deb Hamernik welcomed Clay Lents from the University of Georgia to the group and noted that Matt Cannon from Oregon State University asked to join the project. Everyone was reminded when submitting a Hatch project to make sure assurance statements are valid before submission into the CRIS system. The biosafety and IACUC approval dates must be valid or the submission will be returned to the Experiment Station. Deb described some of the legislative activity that may affect extramural research funding through the Farm Bill. There is a proposal to merge ARS and CSREES administration offices in Washington DC to eliminate administrative duplication. The CREATE-21 and NIFA programs have been proposed in an attempt to double the funding for agricultural research. In the USDA Secretarys proposed 2007 Farm Bill, new funding is proposed for specialty crops, bioenergy/bioproducts, and organic agriculture (primarily crops); however, no new funding for animal production was proposed. The budget for fiscal year 2007 was unusual because Congress did not pass the budget until after 10/01/2006. Therefore the same amount of money was appropriated in 2007 as 2006. Agricultural earmarks were eliminated from the USDA FY2007 appropriation. In FY2007, only the Department of Homeland Security or the Department of Defense received earmarks. The NRI received a small increase in budget in 2007 and expects a similar small increase in 2008. All NRI grants require electronic submission. Applications to NRI must adhere to the published guidelines (budget caps, pages limits, etc.) or they will be returned without review. All NRI submissions are electronic using the SF424 R&R application forms available on grants.gov. Deb also updated personnel changes within USDA CSREES. Deb was questioned about the size of the awards for NRI grants and she answered that the agency is targeting approximately $100,000 (direct costs) per year. Last, Deb updated the group about the bovine and porcine genome sequencing projects. The cattle genome project is starting the annotation phase of the project and is interested in having scientists join the project to link function with gene prediction. The pig genome project will start annotation next year. Business Andrea Cupp explained the rotation and responsibilities of the member at large and the chair of the group prior to asking for nominations for the member at large. Mike Day was elected member at large by a unanimous vote. Old business Tod Hansen explained attempts to coordinate the W1112 meeting with the BIF meeting in Fort Collins. The purpose of having the meetings at the same location would provide members of W1112 to present a program on fetal development as an outreach activity. Support from local extension personnel was positive, however, the idea did not get support from BIF organizers so the attempt was dropped. The group expressed appreciation to Tod and Russ Anthony for the effort. Although the symposium with BIF did not work out, several members of the group said they enjoyed holding the meeting at a station. Dennis Halford presented an update on his work with the Food and Drug Administration to provide sheep tissues as part of the process for CIDR approval in sheep. The tissues have been submitted to the FDA for analysis. Announcements and news from individual stations were given. Individual state announcements occurred with members talking about events, job openings, etc that had happened since our last meeting. New business Laramie, Wyoming was selected as the site for the 2008 W1112 meeting. The meeting will follow the Western Section of ASAS meeting on June 24-26. The W1112 meeting will start in the afternoon on June 26 and finish on June 27. Gary Moss agreed to serve as the local host for the meeting. A comment was made that in the future the group try to have the W1112 meeting in late May to avoid conflicts with meetings that occur later in the summer. Late May 2008 was not an option due to conflict with the 2008 SSR meeting. The group continued a discussion from previous years about ways to increase the exposure of the multi-state project to extension personnel or cattle producers. It was suggested that one or two members of the W1112 group participate in the Beef Symposium during the Western Section ASAS meeting in 2008 in Laramie. This would need to be coordinated with the WSASAS Program Committee. Dennis Halford volunteered to discuss this at the 2007 WSASAS meeting and request to be on the agenda for the 2008 meeting. Milan Shipka volunteered to lead a subcommittee to work with WSASAS to develop a symposium or incorporate W1112 members into a symposium at the WSASAS meeting every 3rd year. Several other ways to improve outreach were mentioned including having a symposium and inviting extension specialists to highlight management research, interaction with the Beef Reproduction Task Force and to have a symposium at the WSASAS meeting every third year to highlight research. Colin read the outreach section of the W1112 project, which is available on NIMS, to the group and emphasized that each member of the group already does the type of outreach described in the project. He suggested that each member include these activities in their report. Dean Hawkins suggested a W1112 poster be made that could be sent to members and used at meetings. A new poster could be made each year and the each new poster would highlight a different objective of the project. The group agreed that a poster should be made and Derek McLean and Dean Hawkins will work on the poster. Members were asked to submit material for the poster. Similar to this idea, Milan suggested that when members do a presentation as an outreach activity, they share the powerpoint file with the other members of the group. The group discussed the use of the W1112 website as a way to share information and powerpoint presentations. It was suggested that in the future a My Space or Sharepoint type of format be used for the website so each station can submit materials including research and teaching resources. Jim Berardinelli updated the group on the website and said the website would need to be set up so that each member can sign in before submitting material to keep the site secure. Jim will work with the information technology person at MSU to update the website. The remainder of the meeting was dedicated to presentations of individual reports, and identification of collaborative projects for the ensuing year. The meeting adjourned at 11:00 am on June 6, 2007. Collaborative Efforts: Collaborators: Nebraska, Wyoming Project: Analysis of testes from rams treated with RU486 from 4-8 weeks. Project: Evaluate behavior, size, and VEGF expression in MGA prepubertal bulls treated with RU486 at 4-8 weeks. Project: Evaluate VEGF isoforms in tissues from fat ewes and malnourished ewes. Collaborators: Montana State, Fort Keogh (Miles City, USDA) Project: Evaluate breeding performance of primiparous suckled beef cows exposed to bulls using a CID7 estrous synchronization protocol and TAI. Collaborators: Arizona, New Mexico Project: Utilization of sheep for research projects Collaborators: Montana, New Mexico Project: Cytology studies of main olfactory epithelium, neuro-sensory epithlia, and mucal odorant binding proteins in pre-ovulatory beef heifers Collaborators: Colorado, Minnesota Project: Fetal tissue collection for steroid transfer Collaborators: Washington, Minnesota Project: Inhibin assay of bovine grafts in mice Collaborators: Wyoming, Minnesota Project: Fertility testing of inhibin immunized ram lambs Collaborators: Colorado, Hawaii Project: Utilizing mRNA for LH receptors in the ovine corpus luteum and effects of PGE, PGE2, and endocannibinoid receptor agonists for CB1 and CB2 receptors in vivo in ewes on luteal function. Collaborators: Hawaii, Texas Project: Effects of endocannibinoid receptor agonist for CB1 and CB2 receptors on bovine luteal function in vitro. Collaborators: Alaska, Colorado Project: Anti-GnRH in reindeer bulls Collaborators: Alaska, Montana Project: Mid-cycle bull exposure and cortisol levels Collaborators: Georgia, Texas, Wyoming Project: Study the influence of feed efficiency on beef cow reproduction Collaborators: Arizona, Nebraska Project: Effect of IGR on testis development Collaborators: Arizona, Michigan Project: Impact of IUGR on ovaries Collaborators: Kansas, Nebraska Project: Evaluate FSH receptors in testes and ovaries during development Project: Quantitation of FSHR3 and FSHR1 at different stages of embryonic development in male and female rats. Collaborators: Nebraska, Washington Project: Effects of inhibitory VEGF isoforms, VEGF, and GDNF on spermatogenesis Collaborators: Michigan, Kansas Project: Evaluate CART signaling in stable cell lines over expressing FSH Receptors 1 and 3. Project: Sharing cell lines. Look at FSHR1 and FSHR3 expression and signaling at different stages of follicular development Collaborators: Michigan, Washington Project: Search for CART and JY-1 mRNA expressions in bull testis tissue. Collaborators: Kansas, New Mexico Project: Effects of energy and protein on expression of FSH-R variants at different stages of follicular development. Collaborators: Washington, Kansas Project: Differences in bovine and ovine FSHR-3 and FSHR-1 during development Collaborators: Wyoming, Arizona Project: Islet isolation and function in vitro Collaborators: Colorado, Wyoming Project: Determining fetal response to maternal infection with BVDV. Collaborators: Colorado, Texas Project: Determining early pregnancy in deer Collaborators: Colorado, The Pennsylvania State University Project: Optimizing pregnancy detection in cattle. Collaborators: Colorado, The Ohio State University Project: impact of reproductive steroid hormone concentrations on uterine mechanisms regulating embryonic survival in cattle Collaborators: WSU, USDA (Miles City). Project. Determine differences in pituitary gene expression in intact, LHRH immunized and castrated bulls.

Objective 1. Investigate molecular, cellular and endocrine mechanisms that limit or control reproductive efficiency in domestic animals. Estrous cycle Physiology/ CL function 1. Both mRNA for nitric oxide and polyamine synthetic enzymes and corresponding product or enzyme activity fluctuate during the ovine luteal phase and in response to PGF2±. 2. Nitric oxide is not luteolytic when infused chronically intrauterine in ewes, but instead may be luteotropic or antiluteolytic via alteration of the PGE:PGF2± ratio. 3. Endothelin-1 (ET-1) is not luteolytic in ewes, but instead may be luteotropic or antiluteolytic via alteration of the PGE: PGF2± ratio in inferior vena cava plasma. Hypothalamic-Anterior Pituitary Gland Physiology 1. The non-genomic effect of estradiol (E2)-BSA, delayed the beginning and the length of the massive release of LH, probably by targeting only the pituitary gland. However it was not able to obliterate the robust effects of E2 induced by acting at both, the hypothalamus and the pituitary gland. 2. Estradiol-BSA stimulated an increase in the number of GnRH Receptors (GnRHR) in cultured ovine pituitary cells comparable to E2 and that stimulation of GnRHR was mediated by a membrane action. 3. The ovine membrane progesterone receptor (mPR) specifically binds progestins as only progesterone, 20±-hydroxyprogesterone and 17±-hydroxyprogesterone significantly displaced binding of 3H-Progesterone to membrane fractions from CHO cells expressing ovine mPR. 4. Tandem linkage of the genes encoding the a, FSHb, and LHb subunits is leading to the construction and expression of an ovine single chain triple-domain chimeric gonadotropin. The ability to construct dually active single-chain analogs could permit more effective control of the half-life and duration of gonadotropin actions. This should aid in the design of analogs that could be used to promote superovulation and/or out-of-season breeding in sheep. Ovarian Physiology 1. The CART peptide is a potent inhibitor of LH stimulated androstenedione production by thecal tissue collected during the early stages of a follicular wave in cattle. These results further support a stage specific local role for CART in regulation of follicular steroidogenesis. 2. The inhibitory effects of CART on FSH induced estradiol production are potentially mediated via regulation of ERK1/2 and Akt activation and (or) activity. CART treatment stimulates expression/activation of phosphatases that in turn terminate FSH-induced Erk1/2 signaling. CART-induced acceleration of dephosphorylation of pErk1/2 and termination of the FSH-induced Erk signaling pathway is mediated, at least in part, by increased tyrosine phosphatase and serine/threonine phosphatase activity that is transcription and translation dependent. 3. Alternative Expression of FSH Receptor Transcripts: We have previously determined that alternate transcripts of the FSH receptor are produced differently at important stages of follicular development in the cow. To better understand the precise role of each form of the FSH receptor on granulosa cell differentiation and development, stable granulosa cell lines have been obtained with produce either FSHR-1 or FSHR-3, and no other forms of the FSHR. Using these cell lines, a series of experiments is planned to characterize patterns of expression of both mRNA and protein for each of these forms of the FSHR, as well as to validate techniques to enhance our ability to detect each form of the FSHR from bovine tissues or primary cultures of granulosa cells. 4. Luteolysis in the ewe appears to involve two stages. Prostaglandin F2a (PGF2a)) decreases progesterone production and an influx of calcium initiates apoptosis. Oxytocin induces an increase in intracellular calcium levels in small luteal cells that may lead to apoptotic cell death, that normal luteal concentrations of progesterone prevent the oxytocin and perhaps the PGF2a -induced increase in the number of small and large luteal cells which respond to these hormones with increased intracellular levels of calcium, and that intraluteal secretion of PGF2a was required for structural luteolysis although steroidogenesis was decreased in the corpus luteum. 5. VEGFxxxb antibody may have greater effects at stimulating follicle progression than treatment with either angiogenic isoform VEGF164 or VEGF120. Higher doses of VEGFxxxb antibody are needed in the ovary in E13 organ cultures to have effects on vascular density when compared to the testis. This would make sense since there are greater amounts of VEGF164b in the ovary. 6. VEGFR-TKI signal transduction inhibitor (inhibits FLT1 and KDR receptors) inhibited both vascular density and follicle progression while the KDR specific inhibitor, V1, inhibited only follicle progression. 7. VEGF188 is greater during late embryonic ovarian development (E16), while the VEGF120 and 164 angiogenic isoforms were greater after birth- P0 to P3 at the time that primordial follicles are forming and progressing to later stages of development. Expression of VEGFxxxb isoforms are in oogonial clusters, and later during ovarian development in all stages of follicles in pre-granulosa, granulosa and theca cells and in vasculature. 8. We have determined there is sex-specific expression of VEGF164b with 4 fold higher levels in the ovary during sex-differentiation than in the testis through real-time PCR for VEGF164b. We believe increased expression in the ovary inhibits endothelial cell migration and allows for vascular to develop from the ovary. While in the testis lower levels of VEGF164b allows for endothelial cell migration and development of vasculature from the adjacent mesonephros. 9. Ovulation of smaller ovulatory follicles results in lower pregnancy rates in beef cows and heifers. A portion of this reduction in fertility appears to be due to reduced subsequent progesterone synthesis. Oocyte-embryo Physiology. 1. JY-1, a novel bovine oocyte/embryo specific protein can regulate function of ovarian somatic/granulosa cells and is required for early embryogenesis. 2. Follistatin treatment of early bovine embryos accelerates time to first cleavage and enhances blastocyst development and cell allocation in favor of trophectoderm cells. Testicular Physiology 1. Immunoneutralization of inhibin holds potential as an approach to increase sperm production. Results show that mean daily sperm production per gram testis was increased 26%. Results also indicated that active immunization against inhibin altered testes weight. Total sperm production was more than doubled in those ram lambs that had heavier testis. Evidence did not support the possibility that LH-stimulated testosterone secretion was decreased following immunoneutralization of inhibin. 2. The adjuvant Emulsigen with the addition of bacterial DNA as an immunostimulant may promote a greater immune response than Freunds adjuvant in bulls. The enhanced immune response may be advantages in the immunoneurtralization of reproductive hormones. Immunization of bulls against a multi-antigenic peptide (MAP) did not increase the immune response over that of a peptide-carrier protein conjugate. 3. Treatment of E13 rat testes with VEGF164 and VEGF120 increased vascular density by 57% (P < 0.05) and perturbed seminiferous cords. 4. We determined that an antibody to the VEGFxxxb isoforms increases vascular density by approximately 100% in the testis and also perturbs cord formation thus; inhibiting VEGFb isoforms may be more affective than adding angiogenic isoforms. 5. VEGFxxxb isoforms are expressed in sub populations of germ cells during early testis development and later are expressed in Sertoli, and interstitial cells. 6. We have injected VEGFxxxb antibody into KDR/LacZ male mice that are adult and prepubertal and are evaluating effects on spermatogenesis. Preliminary data suggest that VEGFxxxb antibody treatment (which prevents actions of VEGFxxxb isoforms) destroys spermatogenesis in certain stage seminiferous tubules. 7. Determined protein expression of VEGF protein with a pan-VEGF antibody (both angiogenic and anti- or inhibitory isoforms) during testis development. All three VEGF isoforms- 120, 164 and 188 are present during cord formation. 8. VEGF has a functional role in promoting germ cell survival and differentiation in bovine testis. Uterine-Placental-Fetal-Nutritional Interactions during Pregnancy. 1. Investigation of pancreatic endocrine formation in sheep determined that mature beta cell replication was occurring concurrently with endocrine differentiation in the fetal pancreas. The highest rates of beta cell proliferation were found during the first half of gestation and then declined to a stable rate in the final trimester. Our data in sheep show that beta cell differentiation and proliferation occur simultaneously during pancreas development. 2. Fetal adaptations of glucose utilization (GUR), insulin action, and glucose production (GPR) in IUGR fetal sheep were investigated. The results indicate that insulin responsive fetal tissues (liver and skeletal muscle) adapt to the hypoglycemic-hypoinsulinemic IUGR environment by developing mechanisms that promote glucose utilization, particularly for glucose storage, including increased insulin action, glucose production, shunting of glucose utilization to glycogen production, and maintenance of glucose transporter concentrations. 3. Sheep fetuses with hyperthermia-induced PI-IUGR are hypoxic, hypoglycemic, hypoinsulinemic and fetal oxygen content is negatively associated with norepinephrine (NE) concentrations. Catecholamines increase with fetal hypoxia and inhibit pancreatic beta-cell insulin release in PI-IUGR fetus. 4. Uterine venous catheters, maternal femoral artery catheters, and uterine artery flow probes were used to investigate early placental development in fetal growth restricted sheep pregnancies. Animals were assigned as either thermoneutral (TN) controls or to hyperthermic (HT) conditions. The data collected so far indicate that while there may be some reduction in uterine blood flow during the first 15 days on treatment, this is countered by elevated maternal arterial pO2 such that oxygen uptake by the uterus is increased. 5. Vascular endotheial growth factor (VEGF) expression is upregulated in the placenta and may not be mediated through the Hif-1 pathway. A 5x5 microarray experiment identified other pathways that may result in induction of angiogenic growth factor expression. 6. Alterations in maternal endocrine status may have influenced maternal transport of nutrients to the developing fetus, resulting in the observed fetal growth restriction. Further, mammary gland wt may have been impacted by maternal endocrine patterns during pregnancy. 7. It appears that nutritional restriction any time during mid- to late pregnancy impacts placentome number, and that cellularity of the placentome can be altered by Se or mid-gestational diet regardless of nutritional levels during late pregnancy. 8. It appears that placentome types do not differ in vascularity or caruncular or cotyledonary angiogenic factor expression. 9. Fetal intrauterine growth restriction resulting from undernutrition during the first half of gestation in ewes and cows have significant negative health effects on their offspring after birth, including insulin resistance, hyperphagia, obesity, decreased skeletal muscle mass, hypertension and infertility . These health, fertility and body compositional effects occurring in offspring of nutrient restricted mothers are not alleviated even if females are realimented during the second half of gestation, and offspring are born at a normal birth weight. 10. In the ewe, the conversion of placentomes from A to D type increases their ability to delivery nutrients to the fetus, thereby protecting the fetus from intrauterine growth restriction. This conversion normally occurs during late gestation in response to the exponential growth of the fetus, but some animals have the ability to convert their placentomes during early gestation in the face of a maternal stressor such as undernutrition. This ability for early placentomal conversion appears to be related to pre-gestational ewe selection, and thus may be epigenetic in nature. 11. Maternal obesity from conception through mid-gestation results in a marked increase in the size of the fetal pancreas and the numbers of insulin secreting ²-cells that it contains per unit area. Because the ²-cell population is derived from pluripotent cells which are partially differentiated and can under go only a limited number of cell divisions, a premature acceleration of ²-cell mitoses could have detrimental impacts on pancreatic function in later life. Nutritional and metabolic effects in non-pregnant animals. 1. In reindeer, changes in body weight over the entire season were not different between groups bred early and late. Plasma will be analyzed for leptin, IGF-1, GH, progesterone, and estrogens. 2. Genetic markers in reindeer are being identified to associate DNA polymorphisms with milk yield, milk composition and calf growth rate. Milk samples were collected from reindeer cows and analyzed for % protein, % fat and % lactose. All calves were weighed at birth and weekly thereafter until weaning and average daily gain was calculated. 3. Estradiol (E2) clearance is decreased during nutritional restriction in sheep. This suggests that dietary alterations in SHBG expression and/or enterohepatic recirculation are also involved in the decreased E2 clearance during fasting or nutrient restriction. 4. LH pulse frequency was suppressed in fasted relative to fed wethers even when fed and fasted animals had uniform circulating E2 concentrations. This result supports the postulate that energy restriction enhances the negative feedback potency of E2. 5. Changes in estrogen clearance associated with nutritional status markedly affect the dynamic pattern of LH secretion in sheep LH is affected by GnRH produced in the pituitary. 6. Feed restriction during post-weaning development of replacement heifers improved efficiency of feed utilization. However, this improved efficiency was accompanied by a 5% reduction in pregnancy rate. Harvested feed requirements per pregnant heifer were reduced by 22% during the 140-d development period, representing a major potential for cost reduction. Behavior 1. Acclimatization to facilities and protocols for 48 hr is necessary for intensive blood sampling in primiparous, postpartum, suckled beef cows. 2. Bull exposure did not alter mean concentrations of cortisol in cows; however, bull exposure changed temporal patterns of cortisol concentrations by decreasing the frequency, increasing the duration of pulses and synchronizing the pulsatile rhythm of cortisol in anestrous, suckled beef cows. 3. Bulls did not appear to influence systemic glucose and thyroid hormones in primiparous postpartum suckled beef cows. However, it appears that adipose metabolism in postpartum beef cows may be affected by the presence of bulls. 4. Exit velocity can be measured reliably as early as 21 to 24 days after birth. Calf exit velocity is associated with serum cortisol concentrations but not with temperament of the cow. 5. Progesterone appears to facilitate sexual behavior in rams 6. Expression of the progesterone receptor in hypothalami of rams may be important for sexual differentiation of behavior. Objective 2. Develop and improve assisted reproductive technologies to enhance sustainability of production systems for domestic ruminants. Estrus, estrous synchronization and AI 1. Switchback breeding trial of variability of gestation length in reindeer showed that gestation length was negatively correlated with day of breeding for both years indicating earlier bred cows have longer gestation length. In addition, gestation length did not differ for cows carrying female calves compared to cows carrying male calves, calf birth weight did not differ between males and females and there was no significant correlation between dam age and gestation length. 2. The relationship of dams body weight, milk components, and milk energy density to reindeer calf growth rate in early and late calving reindeer showed that calf ADG did not differ between groups. 3. Recently, an intravaginal progestogen product, Eazi-Breed CIDR, was approved for estrus synchronization of beef heifers. Comparison of CIDR to MGA in a shorter-term time-breeding program demonstrated that a greater percentage of heifers were pregnant in the CIDR treatment (55%) than in the MGA treatment (33%). 4. Ultrasound analysis can be used to identify reliably (phenotype) cattle with consistently high follicle numbers and low circulating FSH concentration or low follicle numbers and high circulating FSH concentrations during the follicular waves of estrous cycles and such animals differ in their response to superovulation. 5 Progesterone is rapidly metabolized by ovine liver but is stable in skeletal muscle of sheep during long term freezer storage. These data should allow FDA to finalize a decision on whether to approve CIDR use in sheep. 6. Application of intravaginal progesterone during the early postpartum period in spring lambing ewes failed to induce return to estrus. 7. Use of an intravaginal progesterone insert in prepubertal ewe lambs failed to induce puberty at 6.5 mo of age. 8. The use of CIDR as a progestin source in a 14-d progestin, PGF2a, and timed AI and GnRH estrous synchronization protocol is as effective as the use of MGA to synchronize estrus and generate AI pregnancies in beef heifers. 9. Using a CIDR for 7 d with PG given upon of removal of CIDR yields a superior estrous synchronization response compared to that of using a CIDR for 14 d followed 17-d later by PG. However, both CIDR protocols yield similar and acceptable AI pregnancy rates when combined with TAI and GnRH in primiparous beef cows. 10. MGA was a useful tool for shortening length of the lambing season, and although PG600 enhanced ovulation rate, it had no beneficial effects on ewe productivity. Puberty 1. MGA when fed prior to puberty can affect testis size and alter testosterone production in yearling beef bulls. This increase in size is not an alteration in testis composition but a result of a larger testis. Effects on spermatogenesis need to be conducted; however, this may be a viable way to increase testis size and optimize sperm production in male seedstock. 2. Initial investigation on linseed meal (LSM) supplementation during early lactation does not appear to impact onset of puberty in heifer calves. However, it appears that LSM supplementation may enhance wt gain without influencing BCS of heifer calves. Pregnancy detection and maintenance 1. Maternal viral infection during pregnancy programs fetal development and induces a differential type I interferon response. 2. Pregnancy loss in Isg15 null mice is a maternal phenotype that is manifest during implantation 3. Deletion of Isg15 gene results in differential gene expression and pregnancy loss between 7.5 and 12.5 dpc 4. Periattachment factor (PF), a nuclear protein, was first identified in the bovine conceptus at markedly greater concentrations at d 17.5 than d 15.5 of gestation. The robust increase in PF mRNA during a critical time point in early trophoblast expansion, along with its identification in other expanding/regenerating epithelium, may suggest an important role in conceptus development. 5. Regulation of Receptor transporting protein-4 RTP4 during early pregnancy may contribute to the complex signaling cascade associated with pregnancy recognition in this species. 6. Ovine(o) Mx2 expression, like oMx1, is regulated by IFN tau and is expressed in distinct cells of the uterus compared to oMX1. The functional significance of the different cellular localization between oMx1 and oMx2 is not clear, but may suggest that these related proteins possess distinct cellular functions. 7. The oMX1 promoter and enhancer regions were cloned and characterized. Identifying positive and negative regulatory regions in MX1 promoter may suggest a complex regulation of MX1 during early pregnancy in ruminants. Immunocastration 1. Luteinizing hormone-releasing hormone immunization alters pituitary hormone synthesis and storage in bulls and steers. Synthesis and storage of LH and FSH, as measured by pituitary LH and FSH content and expression of the LH ²-subunit and common ±-subunit genes, was suppressed by LHRH immunization. 2. A Bacterial oligodeoxynucleotide immunostimulant is superior to Freunds Adjuvant for LHRH sterilization vaccine in heifers. Treatment with 3.4 mg ova-LHRH in CpG ODN 2006/w-o per immunization is recommended for LHRH immunization in heifers. 3. A longevity study in male rats shows that immunization with LHRH-ovalbumin fusion protein did not shorten the lifespan of the rats, and in fact may have increased the lifespan by several months. This gives some comfort to the vaccine not being detrimental to other systems other than the reproductive system in the animals body. 4. The use of a LHRH vaccine in white tailed deer achieves 80% contraception after 4 years. Simliarly a single shot porcine zona pellucida (PZP) vaccine in white-tailed deer achieved 80% contraception after 5 years. Disease 1. Evidence for steroid hormone regulation of components of the uterine antiviral defense could lead to the development of more effective strategies to block or treat sexually transmitted viral infections. 2. Pronounced antiviral response in maternal and fetal ncpBVDV infection is accompanied by up regulation of ISG15. 3. Development of a single or multiplexing diagnostic for calves that are persistently infected with BVDV is probable. Animal Growth 1. Calm calves have heavier weaning weights than excitable calves. Calves with medium or high serum protein 24 hours after birth have heavier weaning weights than calves with low serum protein. Calves nursing calm or normal temperaments cows had higher serum protein than calves nursing excitable cows. 2. Hauling stress has negative effects on body composition traits, specifically % intramuscular fat and rib fat. 3. Exit velocity measured at feedlot entry failed to be indicative of feedlot performance of steers. 4. We continue to work with the estrogen and phytoestrogen (SDG from flax) model. Previous data indicate that SDG may be anti-estrogenic with increased lengths of feeding. We have found that lactational exposure to SDG in beef cows does not impact puberty, but may influence growth in heifer calves. Immune Function 1. Excitable calves had lower serum IgA concentrations 24 hours after birth compared with calm calves. Serum IgG and IgM concentrations were negatively correlated with serum cortisol concentrations. These results suggest that temperament can influence passive immunity in the newborn calf. 2. Serum protein at 24 hours of age is a better predictor of weaning weight than plasma protein or serum IgG, IgA or IgM. 3. Temperament and thus increased body temperature of calves can negatively affect the response of calves to vaccination.

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