NE1727: Influence of ovary, uterus, and embryo on pregnancy success in ruminants

(Multistate Research Project)

Status: Inactive/Terminating

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SAES-422 Reports

Annual/Termination Reports:

[07/16/2018] [08/19/2019] [08/13/2020] [11/02/2021] [08/05/2022]

Date of Annual Report: 07/16/2018

Report Information

Annual Meeting Dates: 05/13/2018 - 05/15/2018
Period the Report Covers: 10/01/2017 - 09/30/2018

Participants

Bridges, Phillip (pbrid2@email.uky.edu) Univ Kentucky
Butler, Ron (wrb2@cornell.edu) Cornell University
Cheong, Soon Hon (cheong@cornell.edu) Cornell University
Dailey, Robert (rdailey@wvu.edu) West Virginia Univ
Fissore, Rafael (rfissore@vasci.umass.edu) Univ Massachusetts Amherst
Fortune, Joanne (jf11@cornell.edu) Cornell Univ
Giordano, Julio (jog25@cornell.edu) Cornell Univ
Inskeep, Keith (einskeep@wvu.edu) West Virginia Univ
Keating, Aileen (akeating@iastate.edu) Iowa St Univ
Mathew, Daniel (Daniel.mathew@mail.wvu.edu) West Virginia Univ
Memili, Erdogan (em149@ads.msstate.edu) Mississippi St Univ
Pate, Joy (jlp36@psu.edu) Penn St Univ
Rhoads, Shelly (rhoadsm@vt.edu) Virginia Tech Univ
Townson, Dave (dave.townson@unh.edu) Univ New Hampshire
Tsang, Paul (paul.tsang@unh.edu) Univ New Hampshire
Wiltbank, Milo (wiltbank@wisc.edu) Univ Wisconsin
Wood, Jennifer (jwood5@unl.edu) Univ Nebraska
Yao, Jianbo (jianbo.yao@mail.wvu.edu) West Virginia Univ
Administrator: Thompson, Gary (gat10@psu.edu) Penn St Univ
CREEEs rep: Turzillo, Adele (aturzillo@nifa.usda.gov)

Brief Summary of Minutes

Minutes Attachment:

NE1727 Minutes 2018.pdf

Accomplishments

Identification of the impact that thermal stress has on ovarian signaling is resulting in a mechanistic map being developed, upon which strategies to ameliorate seasonal infertility can be based. We have identified that dairy cows have an amazing capacity to develop tolerance to chronic lipopolysaccharide exposure and that no effect of this exposure on the growing dominant follicle is observed. Molecular signaling protein alterations during the follicular and/or luteal phase from lipopolysaccharide or thermal stress treated gilts have been identified. Determined influence of a physiological stressor, obesity, on the ovarian capacity to respond to a toxic environmental stress. Identified molecular proteins within the oocyte that contribute to viability of the oocyte. We have cultured ovarian oocytes to investigate intra-oocyte mechanisms of heat stress induced infertility. Zinc depletion during preantral development impairs oocyte-somatic cell communication and oocyte growth and competence to complete meiosis. NR5A2 is a regulator of luteal progesterone production and may be translationally regulated by miRNA. Changes in the profile of lipids within the CL indicate potential roles for these small molecules as regulators of immune cells, components of intercellular signaling pathways and cell death. We have shown that bovine granulosa and theca express CCN1, along with αV, αIIb, α6, β1, β3 and β5 integrin subunits.  The suppressive effect of PSI on CCN1 expression in KGN cells was accompanied by a decrease in phosphorylated NF-κB activity, while the suppressive effect of Calphostin C and PSI on CCN1 expression in HGrC1 cells probably did not involve NF-κB. We have determined the effects of Ovsynch manipulation on intraovarian events and pregnancy in multiparous dairy cows. We have manipulated ERK signaling (via growth factors) in bovine granulosa cells (GCs) to gain insight about its role in immune-mediated apoptosis of GCs and selection of bovine follicles. Our results provide new information on the property of ZNFO, which will help further elucidation of the molecular mechanisms involved in ZNFO-dependent transcriptional regulation during maternal- embryonic transition. We determined that feeding rumen-protected methionine to high-producing, multiparous dairy cattle reduced pregnancy loss in multiparous dairy cows. We determined that high energy diets could reduce oocyte quality, leading to lack of fertilization. We have defined two other mechanisms that underlie reduced fertility in animals fed high energy diets, namely high progesterone catabolism and reduced embryo quality. An important practical finding was that loss of body condition score from 21 days before calving until 21 days after calving led to reduced fertility in lactating dairy cows. We have determined that early plane of nutrition impacts the progression of uterine gland development in heifers, and that this outcome may be mediated by changes in the expression of several local controllers of gland development. Our recent findings support the contention that maternal obesity modifies a subset of important mediators of embryonic and extraembryonic development in ovine and porcine conceptuses. We determined that maternal obesity also modifies uterine gene expression during early pregnancy in sheep. We determined that Bos indicus and Bos taurus cattle differ in their ability to produce viable offspring following nutrient restriction during early pregnancy. We have discovered that the cytokine, interleukin-6, improves development of the inner cell mass (ICM) in bovine embryos. We determined that early weaning of beef heifers can have long-term effects on physiology.  Some of these changes have implications for lifetime productivity and may be beneficial in some production systems. We investigated the effects of steroid content of follicle fluid on the cumulus-oocyte complex.  Cumulus cell expansion and gene expression differed with estradiol and progesterone concentrations, but in vitro embryo development to the blastocyst stage did not. We determined that a lack of Ca2+ oscillations is not the main reason of the poor success of ICSI in the bovine. This was demonstrated using both in in vitro matured mouse and bovine oocytes injected with bovine sperm. We found that bovine sperm heads are highly resistant to sperm head decondensation, which compromises the ability of in vitro maturated oocytes to reprogram the sperm nucleus. We identified treatments that abruptly change the metabolic status of bull sperm, enhancing the ability of those sperm upon ICSI to initiate Ca2+ oscillations. This also improved the ability of these sperm to undergo decondensation and support pre-implantation embryo development. We have developed the Short-Resynch protocol. This new reproductive management strategy for re-insemination of lactating dairy cows improves reproductive performance and has other benefits for dairy herd management. Compared to traditional resynchronization strategies widely used on dairy farms, Short-Resynch increased pregnancy rates, increased the fertility of subgroups of sub-fertile cows, while taking advantage of inseminations at detected estrus after a previous insemination. We demonstrated that the Short-Resynch program resulted in similar pregnancy rate than one of the most effective resynchronization of ovulation programs available for dairy farms (i.e., Resynch-25) thorough more inseminations at detected estrus, a short interval between inseminations, and increased P/AI for sub-fertile cows. We found that it is possible to collect up to five CL biopsies 48 h apart on the same CL of lactating dairy cows.  Five serial CL biopsies from the same CL in lactating dairy cows did not reduce circulating concentrations of P4, CL size, or affect blood flow. Serial CL biopsies did not cause CL regression during the period of biopsy collection. We determined that a reproductive management strategy designed to increase insemination of cows in estrus after NPD resulted in similar reproductive performance (time to pregnancy) than a typical resynchronization of ovulation protocol used in dairy farms. Proof of concept studies were performed using in vitro mouse follicles and COCs to understand how TNF and H2O2 signaling impact transcriptional and post-transcriptional mRNA synthesis and stability, respectively.  We have demonstrated that obesity induces ovarian inflammation which in turn increases the abundance of oocyte-specific transcription factors, oocyte-specific growth factors, and maternal effect genes. Current studies are discriminating between transcriptional and post-transcriptional regulation of these candidate mRNAs in primary follicles, secondary follicles, and cumulus-oocyte complexes by acute treatments with TNF and H2O2.  Pathway activation by TNF and H2O2 in follicles and COCs are also being elucidated.  We determined that increased abundance of specific polymorphic alleles in the IGF-I and TNFα genes in Holstein cows are favorably associated with increased conception rate to 1st AI and a shorter calving to conception interval during lactation ie. higher fertility. We found that sperm epigenome (chromatin dynamics) as well as functional genome (macromolecules) are associated with bull fertility, and identified key sperm and seminal plasma metabolites that are associated with bull fertility.

Publications

Peer-reviewed journal articles published in 2017 reporting research from this project Hale, B.J., Hager, C.J., Siebert, J.T., Selsby, J.T., Baumgard, L.H., Keating, A.F., Ross, J.W. 2017. Heat stress induces autophagy in pig ovaries during follicular development. Biology of Reproduction. 97(3):426-437. Kvidera, S.K., Horst, E.A., Sanz-Fernandez, M.V., Abuajamieh, M., Ganesan, S., Gorden, P.J., Green, H.B., Schoenberg, K.M., Trout, W.E., Keating, A.F., Baumgard, L.H. 2017. Characterizing effects of feed restriction and glucagon-like peptide 2 administration on biomarkers of inflammation and intestinal morphology. Journal of Dairy Science. 100(11):9402-9417. Ganesan, S., Dickson, M.J., Keating, A.F. Pesticides. In: Encyclopedia of Reproduction. In Press. Ross, J.W., Hale, B.J., Seibert, J.T., Romoser, M. R., Adur, M.K., Keating, A.F., Baumgard, L.H. 2017. Physiological mechanisms through which heat stress compromises reproduction in pigs. Molecular Reproduction and Development.  84(9): 934-945. McNeel AK, Vallet JL, Snelling WM, Wright EC, Larimore EL, Amundson OL, Miles JR, Chase Jr CC, Lents CA, Sonstegard TE, Schroeder SG, Wood JR, Cupp AS, Perry GA, and Cushman RA (2017) Beef heifers with diminished numbers of antral follicles have decreased uterine protein concentrations, Anim Reprod Sci 179:1-9 PMID: 28215453 Romereim SM, Summers AF, Pohlmeier WE, Zhang P, Hou X, Talbott HA, Cushman RA, Wood JR, Davis JS, and Cupp AS (2017) Gene expression profiling of bovine ovarian follicular and luteal cells provides insight into cellular identities and functions, Mol Cell Endocrinol 439:379-394 PMID: 27693538 Romereim SM, Summers AF, Pohlmeier WE, Zhang P, Hou X, Talbott HA, Cushman RA, Wood JR, Davis JS, and Cupp AS (2017) Transcriptomes of bovine ovarian follicular and luteal cells, Data in Brief 10:335-339 PMID: 28004024 Talbott H, Hou X, Qiu F, Guda C, Yu F, Cushman RA, Wood JR, Wang C, Cupp AS, and Davis JS (2017) Early transcriptome responses of the bovine mid-cycle corpus luteum to prostaglandin F2 alpha includes cytokine signaling, Mol Cell Endocrinology 452: 93-109, PMID: 28549990 Talbott H, Hou X, Qiu F, Zhang P, Guda C, Yu F, Cushman RA, Wood JR, Wang C, Cupp AS, and Davis JS (2017) Transcriptomic and bioinformatics analysis of the early time-course of the response to prostaglandin F2 alpha in the bovine corpus luteum, Data in Brief 14:695-706, PMCID: PMC5596332 Wood JR and Cupp AS (2017) Female Endocrinology –Aromatization. Encyclopedia of Reproduction, Second Edition (accepted, in press) Águila L, Felmer R, Arias ME, Navarrete F, Martin-Hidalgo D5, Lee HC, Visconti P, Fissore R. 2017. Defective sperm head decondensation undermines the success of ICSI in the bovine.  Reproduction. 154:207-218. Cerny, K.L., R.A.C. Ribeiro, Q. Li, J.C. Matthews and P.J. Bridges.  2017.  Effect of lipopolysachharide (LPS) on the expression of inflammatory mRNAs and microRNAs in the mouse oviduct.  Reproduction, Fertility and Development.  doi: 10.1071/RD17241. Li, Q., R. Hegge, P.J. Bridges and J. C. Matthews.  2017.  Pituitary genomic expression profiles of steers are altered by grazing of high vs. low endophyte-infected tall fescue forages.  PLoS One.  12(9): e0184612. Willis, E.L., P.J. Bridges and J.E. Fortune.  2017.  Progesterone receptor and prostaglandins mediate LH-induced changes in messenger RNAs for ADAMTS proteases in theca cells of bovine periovulatory follicles.  Molecular Reproduction and Development.  84:55-66. Miles, E.D., B.W. McBride, P.J. Bridges and J.C. Matthews.  2017.  Effect of 17β-estradiol administration on hepatic expression of glutamine synthetase, β-catenin, and GPR30 in aged beef cows.  Canadian Journal of Animal Science.  97:281-289. Hester J, Hanna-Rose W and Diaz FJ.  2017. Zinc depletion reduces fertility and disrupts oocyte development in C. elegans.  Comparative Biochemistry and Physiology 191, 203-209. Tian X, Anthony K and Diaz FJ.  2017. Transition metal chelator induces progesterone production in mouse cumulus-oocyte complexes and corpora lutea. Biological Trace Mineral Research. doi:10.1007/s12011-016-0841-x. Krumm, C. S., S. L. Giesy, L. S. Caixeta, W. R. Butler, H. Sauerwein, J. W. Kim, and Y. R. Boisclair. Effect of hormonal and energy-related factors on plasma adiponectin in transition dairy cows. J Dairy Sci. 100 (11):9418-9427, 2017. Cheong, S. H., O. G. Sa Filho, V. A. Absalón Medina, A. Schneider, W. R. Butler, and R. O. Gilbert. Uterine and systemic inflammation influences ovarian follicular function in postpartum dairy cows. PLoS One 12 (5):e0177356, 2017. Jaskiewicz NJ, Parisi S, Hermawan C, Townson DH 2017 O-GlcNAcylation enhances the tumorigenic properties of cervical cancer cells in vitro. Clinical Obstetrics, Gynecology and Reproductive Medicine 3(3):1-6 Mattos FCSZ, Canavessi AMO, Wiltbank MC, Bastos MR, Lemes AP, Mourao GB, Susin I, Coutinho LL, Sartori R (2017). Investigation of mechanisms involved in regulation of progesterone catabolism using an overfed versus underfed ewe-lamb model. J Anim Sci  95:5537-46. Barletta RV, Maturana M, Carvalho PD, Del Valle TA, Netto AS, Renno FP, Mingoti RD, Gandra JR, Mourao GB, Fricke PM, Sartori R, Madureira EH, Wiltbank MC (2017). Association of changes among body condition score during the transition period with NEFA and BHBA concentrations, milk production, fertility, and health of Holstein cows. Theriogenology 104:30-6. Hackbart KS, Bender RW, Carvalho PD, Vieira LM, Dresch AR, Guenther JN, Gencoglu H, Nascimento AB, Shaver RD, Wiltbank MC (2017). Effects of propylene glycol or elevated luteinizing hormone during follicle development on ovulation, fertilization, and early embryo development. Biol Reprod  97:550-63. Pereira MHC, Wiltbank MC, Guida TG, Lopes FR, Vasconcelos JLM (2017B). Comparison of 2 protocols to increase circulating progesterone concentration before timed artificial insemination in lactating dairy cows with or without elevated body temperature. J Dairy Sci  100:8455-70. Garcia-Guerra A, Motta JCL, Melo LF, Kirkpatrick BW, Wiltbank MC (2017A). Ovulation rate, antral follicle count, and circulating anti-Mullerian hormone in Trio allele carriers, a novel high fecundity bovine genotype. Theriogenology  101:81-90. Garcia-Guerra A, Kirkpatrick BW, Wiltbank MC (2017B). Follicular waves and hormonal profiles during the estrous cycle of carriers and non-carriers of the Trio allele, a major bovine gene for high ovulation and fecundity. Theriogenology  100:100-13. Prata AB, Pontes GCS, Monteiro PLJ, Drum JN, Wiltbank MC, Sartori R (2017). Equine chorionic gonadotropin increases fertility of grazing dairy cows that receive fixed-time artificial insemination in the early but not later postpartum period. Theriogenology  98:36-40. Mollo MR, Monteiro PLJ, Surjus RS, Martins AC, Ramos AF, Mourao GB, Carrijo LHD, Lopes G, Rumpf R, Wiltbank MC, Sartori R (2017). Embryo production in heifers with low or high dry matter intake submitted to superovulation. Theriogenology  92:30-5. Pereira MHC, Sanches CP, Guida TG, Wiltbank MC, Vasconcelos JLM (2017A). Comparison of fertility following use of one versus two intravaginal progesterone inserts in dairy cows without a CL during a synchronization protocol before timed AI or timed embryo transfer. Theriogenology  89:72-8. Baez GM, Trevisol E, Barletta RV, Cardoso BO, Ricci A, Guenther JN, Cummings NE, Wiltbank MC (2017). Proposal of a new model for CL regression or maintenance during pregnancy on the basis of timing of regression of contralateral, accessory CL in pregnant cows. Theriogenology  89:214-25. Sartori R, Spies C, Wiltbank MC (2017). Effects of dry matter and energy intake on quality of oocytes and embryos in ruminants. Reprod Fertil Dev  29:58-65. Ochoa JC, Penagaricano F, Baez GM, Melo LF, Motta JC, Guerra AG, Meidan R, Ferreira JCP, Sartori R, Wiltbank MC (2017). Mechanisms for rescue of CL during pregnancy: Gene expression in bovine CL following intrauterine pulses of Prostaglandins E1 and F2alpha. Biol Reprod DOI:10.1093/biolre/iox183. Toledo MZ, Baez GM, Garcia-Guerra A, Lobos NE, Guenther JN, Trevisol E, Luchini D, Shaver RD, Wiltbank MC (2017). Effect of feeding rumen-protected methionine on productive and reproductive performance of dairy cows. PLoS ONE  12:e0189117. Wijma, M. M. Pérez, M. Masello, D. G. García, M. L. Stangaferro, and J.O. Giordano. 2017. A resynchronization of ovulation program based on ovarian structures present at nonpregnancy diagnosis reduced time to pregnancy in lactating dairy cows. J. Dairy Sci. 101:1697-1707. Wijma, M.L. Stangaferro, M. Masello, G. E. Granados, and J.O. Giordano. 2017. Resynchronization of ovulation protocols for dairy cows including or not including gonadotropin-releasing hormone to induce a new follicular wave: Effects on re-insemination pattern, ovarian responses, and pregnancy outcomes. J. Dairy Sci. 100:7613-7625. S.H. Cheong, O.G. Sá Filho, V.A. Absalon-Medina, A. Schneider, W.R. Butler, R.O. Gilbert. Uterine and systemic inflammation influences ovarian follicular function in postpartum dairy cows. PLoS ONE. 2017 May 12(5):e0177356 doi:https://doi.org/10.1371/journal.pone.0177356. PMID: 28542500 Ealy, A.D. and Wooldridge, L.K. 2017. The evolution of interferon-tau. Reproduction 154:F1-F10. Hughes, C.K., Xie, M.M., McCoski, S.R. and Ealy, A.D. 2017. Activities for leptin in bovine trophoblast cells. Dom. Anim. Endocrinol. 58:84-89. Kelley, D.E., Galvao, K.N., Mortensen, C.J., Risco, C.A. and Ealy, A.D. 2017. Using Doppler ultrasonography on day 34 of pregnancy to predict pregnancy loss in lactating dairy cattle. J. Dairy Sci. 100:3266-3271. Lu, Y., Bradley, J.S., McCoski, S.R., Gonzalez, J.M., Ealy, A.D. and Johnson, S.E. 2017. Reduced skeletal muscle fiber size following caloric restriction is associated with calpain-mediated proteolysis and attenuation of IGF-1 signaling. Am. J. Regul. Integr. Comp. Physiol. 312:R806-R815.  Lugar, D.W., Rhoads, M.L., Clark-Deener, S.G., Callahan, S.R., Revercomb, A.K., Prusa, K.J. and Estienne, M.J. (2017). Immunological castration temporarily reduces testis size and function without long-term effects on libido and sperm quality in boars. Animal 11:643-649. MacGhee, M.E., Bradley, J.S., McCoski, S.R., Reeg, A.M., Ealy, A.D. and Johnson, S.E. 2017.  Plane of nutrition affects growth rate, organ size and skeletal muscle satellite cell activity in newborn calves. J. Anim. Physiol. Anim. Nutr. 101:475-483. Reinholt, B.M., Bradley, J.S., Jacobs, R.D., Ealy, A.D. and Johnson, S.E. 2017. Tissue organization alters gene expression in equine induced trophectoderm cells. Gen. Comp. Endocrinol. 247:174-182.  Talbot, N.C., Sparks, W.O., Phillips, C., Ealy, A.D., Powell, A.M., Caperna, T.J., Garrett, W.M., Donavan, D.M. and Blomberg, L.A. 2017. Bovine trophectoderm cells induced from bovine fibroblasts with induced pluripotent stem cell reprogramming factors. Mol. Reprod. Dev. 84:468-485. Wilson, M.L., McCoski, S.R., Geifer, A.J., Akers, R.M., Johnson, S.E. and Ealy, A.D. 2017. The influence of postnatal nutrition on reproductive tract and endometrial gland development in dairy calves. J. Dairy Sci. 100:3243-3256. Xie, M., McCoski S.R., Johnson, S.E., Rhoads, M.L. and Ealy, A.D. 2017. Combinatorial effects of epidermal growth factor, fibroblast growth factor 2 and insulin-like growth factor 1 on trophoblast cell proliferation and embryogenesis in cattle. Reprod. Fertil. Dev. 29:419-430. Zezeski, A.L., McCracken, V.L., Poole, R.K., Al Naib, A., Smith, J.K., McCann, M.A. and Rhoads ML. (2017) Metabolic and reproductive characteristics of replacement beef heifers subjected to an early weaning regimen involving high-concentrate feeding. Animal 11:820-825.   Conference papers and abstracts/presentations: Yost, E.E., Arzuaga, X., Carlson, L., Keating, A.F., Lehmann, G. 2017. Focusing and refining the evaluation of reproductive endpoints in a systematic review of PCBs.  International Symposium on Systematic Review and Meta-Analysis of Laboratory Animal Studies. Ganesan, S., Nteeba, J. and Keating, A.F.  2017. Impact of glyphosate on ovarian signaling pathways regulating folliculogenesis and steroidogenesis. Society for the Study of Reproduction annual meeting.  Hines, E.A., Romoser, M., Keating, A.F., Baumgard, L.H., Niemi, J., Williams, N.H., Haberl, B., Kerr, B., Touchette, K.T., Ross, J.W. 2017. Supplementation of arginine does not improve gilt reproductive performance under commercial conditions. Society for the Study of Reproduction annual meeting.  Hines, E.A., Romoser, M., Keating, A.F., Baumgard, L.H., Niemi, J., Haberl, B., Kerr, B., Touchette, K., Ross, J.W. 2017. Effect of maternal arginine supplementation on offspring performance of pigs in a commercial production environment.  Importance of nutrition and environment on birth weight, muscle growth, health and survival of the neonate workshop meeting. Baumgard, L.H., Kvidera, S.K., Horst, E.A., Dickson, M.J., Ydstie, J.A., Shouse, C.S., Mayorga, E.J., Al-Qaisi, M., Lei, S., Bidne, K.L., Seibert, J.T., Hall, B.J., Keating, A.F., Ross, J.W., Selsby, J.T., Rhoads, R.P. 2017. Consequences of leaky gut on the immune system, metabolism, physiology and animal performance. American Society of Dairy Science annual meeting. Dickson, M.J., Kvidera, S.K., Horst, E.A., Ydstie, J.A., Bidne, K.L., Wiley, C.E., Gunn, P.J., Keating, A.F., Baumgard, L.H. Chronic lipopolysaccharide infusion has no impact on dominant follicular size but affects 17β-estradiol in lactating dairy cows.  American Society of Dairy Science annual meeting. Kvidera, S.K., Horst, E.A., Sanz Fernandez, M.V., Abuajamieh, M., Ganesa, S., Gordon, P.J., Green, H.B., Schoenberg, K.M., Trout, W.E., Keating, A.F., Baumgard, L.H. 2017. Glucagon-like peptide 2 administration improves biomarkers of inflammation and intestinal morphology in feed restricted lactating Holstein cows.  American Society of Dairy Science annual meeting. Kvidera, S.K., Dickson, M.J., Horst, E.A., Ydstie, J.A., Shouse, C.S., Bidne, K.L., Mayorga, E.J., Al-Qaisi, M., Ramirez, H.A., Keating, A.F., Baumgard, L.H. 2017. Effects of continuous and increasing lipopolysaccharide infusion on basal metabolism in lactating cows.  American Society of Dairy Science annual meeting. Dickson, M.J., Kvidera, S.K., Horst, E.A., Ydstie, J.A., Shouse, C.S., Mayorga, E.J., Al-Qaisi, M., Bidne, K.L., Ramirez, H.A., Keating, A.F., Baumgard, L.H. 2017. Metabolic responses to a glucose tolerance test and epinephrine challenge post-continuous lipopolysaccharide infusion in lactating cows.  American Society of Dairy Science annual meeting. Dickson, M.J., Kvidera, S.K., Horst, E.A., Ydstie, J.A., Shouse, C.S., Mayorga, E.J., Al-Qaisi, M., Bidne, K.L., Ramirez, H.A., Keating, A.F., Baumgard, L.H. 2017. Chronic lipopolysaccharide infusion reduces productivity in lactating dairy cows.  American Society of Dairy Science annual meeting. McCain AR, Beede KA, Yates DT, and Wood JR (2017) Maternal and Paternal Obesity Differentially Affect Fetal Growth with Maternal Obesity Associated Growth Restriction Attributed to Decreased 11-Hydroxysteroid Dehydrogenase Expression Society for the Study of Reproduction, Washington, DC Timme KA, Xie F, Davis JS, and Wood JR (2017) Ovarian Inflammation and Oxidative Stress Associated with Diet Induced Obesity (DIO) Impacts RNA-Binding Protein Expression and Potentially mRNA Stability in the Murine Ovary and Oocyte.  Society for the Study of Reproduction, Washington, DC Romereim SM, Summers AF, Pohlmeier WE, McFee RM, Spuri-Gomes R, Kurz SG, Davis JS, Wood JR, and Cupp AS (2017) A High-Androgen Microenvironment Inhibits Granulosa Cell Proliferation and May Alter Cell Identity. Society for the Study of Reproduction, Washington, DC Abedal-Majed MA, Hart ML, Largen V, Magamage MPS, Kurz SG, Sargent KM, Bergman J, McFee RM, Cushman RA, Davis JS, Wood JR, and Cupp AS (2017) Ovarian Cortex from High A4 Cows Secrete Excess A4 and Exhibits Increased Oxidative Stress, Macrophage Markers and Arrested Follicle Development Which can be Partially Rescued by Angiogenic VEGFA Isoforms. Society for the Study of Reproduction, Washington, DC Nafziger S, Abedal-Majed MA, Tenley S, Summers A, Hart ML, Harsh G, Bergman J, Kurz SG, Wood JR, Cushman RA, and Cupp AS (2017) Endocrine Profiles during Attainment of Puberty may Predict Reproductive Longevity in Heifers. Society for the Study of Reproduction, Washington, DC Wood JR (2017) Maternal Obesity, the Gut Microbiota, and Oocyte mRNAs: Potential Impact on the Developing Embryo and Fetus. 4th World Congress on Reproductive Biology, Naha, Okinawa, Japan (invited speaker) McCain AR, Beede KA, Yates DT, Shankar K, and Wood JR (2017) Maternal Obesity Results in Fetal Growth Restriction Associated with Reduced Placental Efficiency and an Altered Placental Transcriptome. 14th Annual Gilbert Greenwald Symposium on Reproduction, Kansas City, KS Timme KA, Xie F, Davis JS, and Wood JR (2017) Ovarian Inflammation and Oxidative Stress Associated with Diet Induced Obesity (DIO) Impacts RNA-Binding Protein Expression and Potentially mRNA Stability in the Murine Ovary and Oocyte. 14th Annual Gilbert Greenwald Symposium on Reproduction, Kansas City, KS Nafziger S, Abedal-Majed MA, Tenley S, Summers AF, Hart ML, Harsh G, Bergman J, Kurz SG, Wood JR, Cushman RA, and Cupp AS (2017) Endocrine Profiles during Attainment of Puberty may Predict Reproductive Longevity in Heifers. 14th Annual Gilbert Greenwald Symposium on Reproduction, Kansas City, KS Abedal-Majed MA, Hart ML, Largen V, Magamage MPS, Kurz SG, Sargent KM, Bergman J, McFee RM, Cushman RA, Davis JS, Wood JR, and Cupp AS (2017) Ovarian Cortex from High A4 Cows Secrete Excess A4, and Exhibit Increased Oxidative Stress and Arrested Follicle Development which can be Partially Rescued by Angiogenic VEGFA Isoforms. 14th Annual Gilbert Greenwald Symposium on Reproduction, Kansas City, KS Wijma, M. Masello, M. L. Stangaferro, M. M. Pérez, and J. O. Giordano. 2017. A resynchronization of ovulation strategy based on the ovarian structures present at non-pregnancy diagnosis reduced time to pregnancy in lactating dairy cows. J. Dairy Sci. Volume 100, E-Supplement 2. E.M. Sitko, S.H. Cheong 2017 Longitudinal study of metritis risk in dairy cattle. Clinical Theriogenology Sep; 9(3):432. Ardestani G, Mehregan A, Carvacho I, He C, Fissore RA. Expression and Function of Cation Permeable Channels in Mouse GV oocytes. Gordon Research Conferences. July 2017- Fertilization and Activation of Development. New Hampshire. Department of Veterinary and Animal Sciences, University of Massachusetts, Amherst, MA 01003. Huang, P.J. Bridges and J.C. Matthews. 2017. Shifts in hepatic transcriptome profiles of growing versus finished beef steers. Journal of Animal Science 95 (Supplement 4) doi:10.2527/asasann.2017.326. Huang, Y. Jia, Q. Li, W.R. Burris, P.J. Bridges and J.C. Matthews. 2017. GTRAP3-18 protein negatively modulates canalicular glutamate transport and glutamine synthesis capacity in the liver of finishing versus growing beef steers.  Journal of Animal Science 95 (Supplement 4) doi:10.2527/asasann.2017.327. Li, P.J. Bridges and J.C. Matthews. 2017. Summer-long grazing of endophyte-infected tall fescue by growing beef steers inhibits expression of genes responsible for prolactin and ACTH synthesis. ASAS 2017 Southern Section Meeting. doi:10.2527/ssasas2017.058 Jia, Q. Li, G.E. Aiken, P.J. Bridges and J.C. Matthews. 2017. Effects of Selenium-form Phenotypes on Steers Grazing Endophyte-Infected Tall Fescue.  ASAS 2017 Southern Section Meeting. doi:10.2527/ssasas2017.057. Fetter, ME, Pate, JL, Harvatine, KJ, Moats, J and Ott, TL. 2017. Effects of feeding an extruded flaxseed supplement on fatty acids in milk and plasma and immune function in transition dairy cows. Annual Meeting of the American Dairy Science Association, Pittsburg, PA. Hughes CK and Pate JL. 2017. Differentially expressed proteins, transcripts, and miRNA in the corpus luteum during maternal recognition of pregnancy indicate matrix remodeling and miRNA regulation during luteal rescue. Proceedings of the 50th annual meeting of the Society for the Study of Reproduction, Washington, DC. Wetzel L, Inskeep EK and Pate JL. 2017. The importance of the luteal microenvironment for programming resident macrophages. Proceedings of the 50th annual meeting of the Society for the Study of Reproduction, Washington, DC. Pate JL and Hughes CK. 2017. Cell and networks the facilitate luteal survival for pregnancy success. Proceedings of the 4th World Congress on Reproductive Biology, Okinawa, Japan. Pate JL, Maalouf SA and Hughes CK. 2017. MicroRNA as regulators of luteal function. Proceedings of the 37th annual meeting of the American Society of Reproductive Immunology, Chicago. Kutchy NA, Menezes ESB, Wills RW, Tan W, Kaya A, Topper E, Didion BA, Moura AA, Perkins A, Memili E. (2017) Sperm Nuclear Proteins are Associated with Bull Fertility. Presented at the International Conference of Society for the Study of Reproduction. Washington, DC. July 13-16, 2017. Kutchy NA, Menezes E, Moura A, Kaya A, Perkins A, Memili E. (2017) Sperm Epigenomic and Genomic Determinants Regulating Male Fertility. Gordon Research Conference on Fertilization & Activation of Development: Uniting Eggs and Sperm, from Fundamentals to Applications. Holderness, NH July 16-21, 2017. Haynes, BP, Kutchy NA, Menezes ESB, Ugur MR, Tan W, Moura A, Kaya A, Memili E. (2017) Epigenetic Control of Male Fertility through Sperm Histone 4. Undergraduate Research Symposium, Mississippi State University, April 13, 2017. Ugur MR, Kutchy NA, Menezes SBE, Kaya A, Moura A, Perkins A, Memili E. (2017) Sperm- and Oocyte-born MicroRNA Transcripts Regulating Early Mammalian Embryogenesis and Fertility. Mississippi State University’s Graduate Student Association’s Symposium, March 25, 2017. Kutchy NA, Menezes ESB, Chiappetta A, Kaya A, Moura A, Perkins A, Memili E. (2017) Sperm Chromatin Dynamics Modulated by Acetylated and Methylated Sperm Histone 3 (H3K27ac and H3K27me3) are Associated with Male Fertility. Triangle Consortium for Reproductive Biology (TCRB) Conference, Research Triangle Park, NC, February 25, 2017. Koganti PP, Wang J and Yao J. Identification and Functional Prediction of Bovine Oocyte-Specific Long Non-Coding RNAs. 50th Annual Meeting of the Society for the Study of Reproduction. Washington D.C. July 13-16, 2017. Zhang M, Hand JM, Smith GW and Yao J. Determination of the consensus target sequences recognized by ZNFO, a novel oocyte-specific zinc finger transcription factor in cattle. 50th Annual Meeting of the Society for the Study of Reproduction. Washington D.C. July 13-16, 2017. Harl, A.W., E.L. Larimore, A. Al Naib&dagger;, L.K. Wooldridge, A.D. Ealy, G.A. Perry and M.L. Rhoads. 2017. Maturation of bovine cumulus-oocyte complexes with follicle fluid varying in estradiol content affects cumulus cell expansion without affecting subsequent embryo development in vitro. 2017 IETS Annual Meeting, Austin, TX. J.O. Giordano. Integrating automated detection of estrus in reproductive management programs for dairy cattle. Precision Dairy Management Conference. Lexington, KY. May 30th, 2017. J.O. Giordano. Update on reproductive physiology and reproductive management programs of lactating dairy cows. Technical services training program. Boehringer Ingelheim Vetmedica. Portland, OR. June 22nd, 2017. J.O. Giordano. Reproductive Physiology and Management of Dairy Cattle. Alta Genetics Dairy Manager School for Spanish speaking managers. Alta Genetics. Watertown, WI. July 17th and 18th, 2017. J.O. Giordano. Reproductive Physiology and Management of Dairy Cattle. Alta Genetics Dairy Manager School for Spanish speaking managers. Alta Genetics. Tulare, CA. May 23rd, 2017. J.O. Giordano. 1. Practical aspects of reproductive physiology. 2. Current reproductive management strategies for dairy cattle. Eastern New York Dairy Day. Cooperstown, NY. March 28th, 2017. J.O. Giordano. Reproductive Physiology and Management of Dairy Cattle. Alta Genetics Dairy Manager School. Alta Genetics. Gainesville, FL. March 9th, 2017. J.O. Giordano. Reproductive Physiology and Management of Dairy Cattle. Alta Genetics Dairy Manager School. Alta Genetics. Gainesville, FL. February 20th, 2017. J.O. Giordano. Ohio Dairy Health and Management Certificate Program: Dairy Cattle Reproduction with Emphasis on Transition Cow Management in Confinement Systems. 1. Automating Health Monitoring in Dairy Farms. 2. Automating Health Monitoring in Dairy Farms. College of Veterinary Medicine, The Ohio State University, Columbus, Ohio. February 2nd, 2017. J.O. Giordano. Automating Reproductive and Health Management in Dairy Farms. GEA Technologies technical services team training program. Puerto Vallarta, Mexico. January 18th, 2017.   Extension reports: Romereim SM, Tenley SC, Abedal-Majed MA, Bergman JW, Kurz SG, Davis JS, Wood JR, and Cupp AS (2017) Letrozole: A Steroid-Free Estrous Synchronization Method. Nebraska 2017 Beef Cattle Report, in press   Theses/Dissertations: Abedal-Majed, MA (2017) Effect of Post-Weaning Diet, Excess Androstenedione, and Vascular Endothelial Growth Factor A (VEGFA) Isoforms on Follicular Progression in Bovine Ovarian Cortical Cultures (Nebraska). Joseph Miseirvitch (2017). Regulation of CCN1 Expression by PGF and PKC Signaling in Human Ovarian Granulosa Cells (New Hampshire). McCauley Vailes (2017). Post-Transfer Outcomes in Cultured Bovine Embryos Supplemented with Epidermal Growth Factor, Fibroblast Growth Factor 2, and Insulin-Like Growth Factor 1. (Virginia). Benjamin R. Crites (2017).  Comparison of conception rates in beef cattle inseminated with either SEXEDULTRA™ sex-sorted semen or conventional semen in fixed-time artificial insemination (FTAI) protocols (Kentucky). Benjamin Hale (2017). Two components of maintaining developmental competence: MicroRNA-21 in the maturing oocyte and autophagy induction in the follicular stage ovary (Iowa). Katie L. Bidne (2017). Investigating the ovarian response to endotoxemia (Iowa). Mackenzie J. Dickson (2017). Impact of endotoxemia on ovarian signaling and function (Iowa).   Active collaboration within the group NY, PA, NH, VT and WV:  Samples collected to investigate associations between fertility outcomes and SNP in candidate genes from dairy cows. IA and UK: Samples shared to determine the effect of LPS treatment on ovarian inflammation. PA and WV: Samples shared for analysis of molecular regulators of luteal function. VT and NY: Samples assayed (NY) for analysis of progesterone concentrations. NY and WI:  Samples collected to investigate the effect of supplementation with rumen-protected methionine on reproductive traits of lactating dairy cows.   In addition to Hatch Multistate Funds, these studies were supported by: National Institute of Environmental Health Science (AK)      National Institutes of Health (FD) USDA-National Institute of Food and Agriculture (AK, PB, JP, EM, MR, AE, SHC, JG, MW) USDA-ARS Specific Cooperative Agreement (SCA) with the Forage and Animal Production Research Unit (FAPRU), Lexington, KY (PB) Virginia Agricultural Council Research Funding (AE, MR) Iowa Pork Producers Association (AK)                                                                                              National Pork Board (AK) National Institute of Food and Agriculture (AK) Iowa Pork Industry Center (AK) Iowa State University Bailey Career Development Award (AK) Lee Rumberger and Family Endowment (JP) Alta Genetics, Inc. (EM) Brasil Programa de Doutorado Integrado em Zootecnia – PDIZ/UFC and Conselho Nacional de Desenvolvimeno Científico e Tecnológico – CNPq (EM) New York Farm Viability Institute (JG) Adisseo (JG, MW) Zoetis (JG)

Impact Statements

Low pregnancy rates in beef and dairy cows result in increased costs to producers associated with rebreeding or culling animals, and recouping heifer development costs. Reduced fertility is also a major cause of reduced milk (dairy) and meat (beef) production which impacts the food supply. The focus of the NE1727 multistate project “Influence of ovary, uterus, and embryo on pregnancy success in ruminants”, is to understand how altered ovarian function, impaired oocyte quality, and disruptions of conceptus-uterine interactions contribute to infertility, and devise management strategies that will overcome these factors in order to optimize the chances that animals successfully achieve a pregnancy. To achieve this goal, the following three objectives will be performed: (1) Determine the impact of altered ovarian function on reproductive performance; (2) Identify alterations in embryo development and uterine and CL function associated with declining pregnancy establishment; and (3) Identify changes in genetics and reproductive management that lead to improved pregnancy rates. As investigators in NE1727, we will use our collective expertise to understand the mechanisms by which internal and external forces modify these important physiological processes and to use this information about core biological processes to rationally develop new and innovative tools to increase cyclicity and breeding efficiency, and reduce early embryonic failures, thereby improving reproductive efficiency of both dairy and beef cattle operations in the United States.

Date of Annual Report: 08/19/2019

Report Information

Annual Meeting Dates: 05/20/2019 - 05/21/2019
Period the Report Covers: 10/01/2018 - 09/30/2019

Participants

Phil Bridges
Ben Crites
Kasey Schalich
Sarah Carr
Bob Daly
Dan Mathew
Jianbo Yao
Milo Wiltbank
Julio Giordano
Alan Ealy
Joy Pate
Camilla
Erdogan Memili
Dan Poole
Jen Wood
Aileen Keating
Shelly Rhoads
Ron Butler
Paul Tsang
Michael Goulet
Gary Thompson
Cristina Soriano-Ubeda

Participants Attachment:

NE1727 Annual Report 2018.pdf

Brief Summary of Minutes

Minutes pasted with participant informatio.

Accomplishments

The following have been accomplished:   Identification of the impact that thermal stress has on ovarian signaling is resulting in a mechanistic map being developed, upon which strategies to ameliorate seasonal infertility can be based.   Determined influence of a physiological stressor, obesity, on the ovarian capacity to respond to a toxic environmental stress.   Identified molecular proteins within the oocyte that contribute to viability of the oocyte.   Characterization of ovarian connexin proteins 37 and 43 to understand communication between somatic cells and the female gamete.   We have cultured ovarian oocytes to investigate intra-oocyte mechanisms of heat stress induced infertility.   Identified ovarian proteomic effects of obesity.   Determined that the response of OVCAR8 cells to PGF2α and PMA are not as robust as the human granulosa tumor cell line, KGN. While protein kinase C may have a role in regulating CCN1 expression, the constitutively high expression of STAT3 may be inhibiting CCN1 in OVCAR8 cells.   Determined that LH and mediators of the protein kinase A pathway, forskolin and db-cAMP, generally had no effect on CCN1, VEGFA and FGF2 expression in day 4 and 8 luteal cells. Investigated the effects of Ovsynch manipulation on intraovarian events and pregnancy in multiparous dairy cows.   Examined the effects of location of the presumptive ovulatory follicle relative to the regressing CL (ipsilateral vs. contralateral).   Determined that TGFα stimulates proliferation of bGCs from small antral follicles via a MAPK-mediated mechanism, but may be incapable of promoting their persistence in follicles during the process of follicular selection/dominance.   Determined that the transcription factor, NR5A2, facilitates progesterone production in luteal cells, is inhibited by PGF2A, and is regulated by at least three microRNA, miR-27b-3p, miR-432-5p and miR-369-3p.   Completed metabolomic analyses that suggested a role for lipid mediators during luteal development and early pregnancy, as regulators of steroidogenesis, immune cells, intracellular signaling, and cell death. Two lipids that have been overlooked in the CL but may be important regulators of luteal function are 15-KETE and 5-KETE.   Discovered that conceptus signals alter the proportions and functions of immune cells in the peripheral blood and the fetal-maternal interface resulting in increased expression of functions associated with immune tolerance during early pregnancy in cattle.   Determined that zinc depletion impairs germ cell nest breakdown and follicle activation suggesting.   Identified potentially novel signaling pathways involved in oocyte maturaiton and cumulus expansion.   Determined that GHR and IGFI SNPs had no significant effect on fertility.   The SNP in TNFα had significant effects on fertility; BB cows being most disadvantaged.   Neither IGFI nor TNFα SNPs had a linear effect on days to pregnancy. However, combining the effects of SNPs in TNFα + IGFI, cows carrying more favorable alleles (AA for each gene) had fewer DIM to pregnancy than cows having BB genotype for both genes (AAAA=112±8, BBBB=126±10; Linear effect P=0.019).   Using a new dataset with 84 multiparous lactating dairy cows, genotyping has been conducted on SNP’s in another candidate gene: Coenzyme Q9 (COQ9).   Determined that cumulus cell expansion and gene expression of in vitro matured oocytes differ based upon estradiol and progesterone concentrations in maturation media.  Subsequent in vitro embryo development to the blastocyst stage was not affected by those treatments applied during the maturation phase.    Determined that the addition of progesterone to maturation media containing follicle fluid supports embryo development to the blastocyst stage, while the addition of estradiol (alone or in combination with progesterone) inhibits blastocyst development.   Determined that maternal obesity modifies a subset of important mediators of embryonic and extraembryonic development in ovine conceptuses. Maternal obesity also modifies uterine gene expression during early pregnancy in sheep.   Volatile fatty acids can be used to manipulate progesterone profiles in cattle.   Investigated Bos indicus and Bos taurus cattle and how they differ in their ability to produce viable offspring following nutrient restriction during early pregnancy.   Determined that the cytokine, interleukin-6, improves development of the inner cell mass (ICM) in bovine embryos by utilizing STAT3-dependent signals.   Supplementing in vitro-produced bovine embryos will selective uterine-derived bioactive factors can modify events of early pregnancy after embryo transfer.   Determined that epidural administration of GnRH to cattle causes ovulation, but does not shorten the time to ovulation or lessen the variability in time to ovulation. While previous studies indicated this may be a strategy to improve ovulatory response (occurrence and timing), the current studies conducted in beef cows and heifers did not prove this hypothesis.   We have characterized the physiology that produces the high ovulation rate in carriers of Trio, a bovine high fecundity allele that leads to overexpression of SMAD6 in granulosa cells. Multiple follicles are selected in Trio at a smaller size, but at a similar time after initiation of a follicular wave, and with similar changes in granulosa cell gene expression as single-ovulating, non-carrier sisters (increased LH receptor, aromatase, PAPPA).   Determined that trio carriers have similar primordial follicle numbers and circulating AMH as non-carriers but an elevation in circulating FSH near the time of deviation, likely a key contributor to the increased ovulation rate in Trio carriers.   Discovered that treatment with a GnRH antagonist (Acyline) suppresses LH pulses and blocks selection of a dominant follicle and follicle growth past 8.5 mm. Provision of hCG (LH activity) restores dominant follicle growth. Thus, growth of follicles prior to 8.5 mm is not dependent on LH pulses but selection of a dominant follicle and growth after 8.5 mm is dependent on LH pulses.   The follicle selection process can be categorized by the size of the subordinate follicle at the time of selection into four categories: Conventional, undersized, Co-dominance, or Switched dominance. In elevated circulating progesterone heifers primarily undergo undersized deviation, whereas conventional deviation occurs in low progesterone concentrations.   Determined that although circulating LH is decreased in high progesterone, provision of hCG does not increase conventional deviation but tended to decrease the subordinate follicle size at deviation. Thus, undersized deviation is not due to suppressed circulating LH.   We determined the patterns of prostaglandin F2α (PGF) pulses using circulating PGF metabolite (PGFM) during luteolysis, early pregnancy (Day 18-21), and the second month of pregnancy. Luteolysis is associated with distinct PGFM pulses that are suppressed during early pregnancy but are present during later pregnancy. Apparently mechanisms maintaining the CL are different in early and later pregnancy.   We have determined the differentially expressed (DE) transcriptome (RNASeq) during early pregnancy compared to cows at a similar stage of non-pregnancy that have not been exposed to prostaglandin F2α (PGF) pulses (60 upregulated and 27 downregulated DE genes). Most of the upregulated genes were related to interferon-stimulated pathways.   We have determined the DE transcriptome during early luteolysis (initial exposure to PGF pulses but no decrease in circulating P4; 173 DE genes) and later luteolysis (after exposure to PGF metabolite > 100 pg/mL; 4615 DE genes).   We have used the RNASeq data from all the different physiologic states to identify and compare stably transcribed genes for future use as house-keeping genes in the CL.   A cytobrush method was validated for sequential analysis of gene expression in the endometrium before, during, and after luteolysis in heifers with 2 or 3 follicular waves.   Discovered that endometrial mRNA for progesterone receptors and estradiol receptors 1 and 2 were greater on Day 8 and decreased thereafter similarly in heifers with two and three follicular waves. Oxytocin receptor mRNA increased earlier in two vs three-wave cycles (Day 14 vs 18) and the increase was associated with the onset of luteolysis. Thus, endometrial expression of mRNA for the steroid receptors did not explain the variability in timing of onset of luteolysis in heifers while the later onset of luteolysis in three-wave cycles was associated with later up-regulation of oxytocin receptor mRNA.   Using a physiologic model of the the proestrous period, it was found that the changes in endometrial area normally associated with estrus only occurred with both a decrease in circulating P4 and an increase in circulating estradiol.   Endometrial expression of the receptor for oxytocin increases on Day 18 in non-pregnant heifers, prior to luteolysis, and on Day 32 of pregnancy but is suppressed on Day 18 of pregnancy, probably by interferon-tau.   Determined that endometrial expression of interferon-stimulated genes (ISGs) were elevated in Day 18 pregnant heifers but were low on Day 18 in non-pregnant heifers or on Day 32 of pregnancy.  Objective 3: Identify changes in genetics and reproductive management that lead to improved pregnancy rates in ruminants.   Determined that production of an accessory CL using GnRH or hCG decreases pregnancy loss in recipients of in vitro produced (IVP) embryos.  In one study (n = 552), heifers were treated with 2,000 IU of hCG or were not treated after either AI or at time of transfer of an IVP embryo. Treatment with hCG caused ovulation and increased circulating P4 in both groups but only decreased pregnancy loss in IVP recipients and not in heifers that received AI.   In another study (n = 1,562), heifers were treated on Day 5 with 200 ug GnRH or were not treated and received an IVP embryo on Day 7 (+ 1 d). Treatment with GnRH caused ovulation (83.9%), increased circulating P4, and decreased pregnancy loss between Day 33 and 60 of pregnancy. Pregnancy loss between day 33 and 60 in recipient heifers with or without an accessory CL present on D33 in an IVP-ET program.   Determined that presence of an accessory CL at the D33 pregnancy diagnosis was associated with a dramatic reduction in pregnancy loss from D33 to D60 in recipients of Stage 7 (expanded blastocyst) embryos (11.6 vs 27.6%).  Thus, formation of an accessory CL using hCG or GnRH does not seem to alter pregnancy per embryo transfer at the Day 33 pregnancy diagnosis but decreases pregnancy loss between Day 33 and Day 60 in recipients of Stage 7 embryos.     Cows were classified as Control or High A4 based on detection of A4 in culture medium from ovarian cortex.  Furthermore, monthly blood and fecal samples were collected from beef heifers (n=112/year) beginning at weaning (Oct 2017, 2018) until breeding or ovariectomy (June 2018, 2019).  In addition, a sample was collected from each heifer just prior to weaning.   Blood samples from Control or High A4 cows were assayed for progesterone (P4) and heifers were classified into one of four puberty groups: typical (control), early, start-stop, or noncycling. DNA will be collected from fecal samples and metagenomic (shotgun sequencing) analyses performed on Control (n=18) and High A4 (n=18) cows.    We have collected DNA from 3 Control and 3 High A4 cows which were sequenced in 2018.  Additional samples (6 per experimental group) will be sequenced in 2019 and bioinformatics analyses initiated.  We are also collecting monthly fecal samples from heifers between weaning and breeding which are classified based on days of age a puberty attainment and continued cyclicity; typical (378 ± 2 days), early 317 ± 4 days), start-stop (265 ± 4 days but discontinued cyclicity), and noncycling (no progesterone > 1 ng/mL).   Two new studies were initiated this year to measure the effect of inflammation on theca cell steroidogenesis.   Cytokine arrays showed increased TNFα and IL-1β secreted from theca cells from High A4 compared to control cows. We are in the process of measuring the concentration of LPS in follicular fluid collected from High A4 and control cows.   We have initiated a study, to localize lipids including cholesterol to specific cell types in the ovary.  The technique, Mass-Spectrometry Imaging, can both localize and determine the relative amount of specific lipid species in granulosa and theca cells from High A4 and Control cows.   A population of cows with increased ovarian androstenedione (High A4; >40ng/mL) concentrations has been identified in the UNL physiology herd. Interestingly, circulating and follicular fluid pro-inflammatory cytokine composition is altered in High A4 cows suggesting chronic inflammation.   Estrous cycles of High A4 and Control cows were synchronized and stimulated for 3 days with FSH (210 IU) prior to ovariectomy. Theca cells were micro-dissected from follicles >7mm and immediately cultured in untreated medium for 120 minutes or LPS-containing medium (50ng/ml) for 15 to 120 minutes. Western blot analysis showed progressive LPS-dependent increases in MyD88 in theca cells from Control cows. However, MyD88 expression was progressively decreased in theca cells from High A4 cows. Cytokine antibody arrays showed significant increases in pro-inflammatory cytokine concentrations (TNFα, IL-1α, and IL-21) in untreated medium from High A4 compared to Control theca cells. When Control theca cells were treated with LPS, TNF, IL-1, and IL-21 were significantly increased in a time-dependent manner.    Determined that LPS treatment of High A4 theca cells resulted in significantly decreased levels of cytokines. These results suggest insensitivity of High A4 theca cells to LPS-mediated inflammatory response which may be due to chronic in vivo LPS exposure. Previous studies show that LPS increases ovarian steroidogenesis suggesting that chronic exposure of theca cells to LPS may be a mechanism for androgen excess in High A4 cows.   A population of cows that have excess androstenedione (A4; High A4) in follicular fluid of dominant follicles and secreted from ovarian cortex media (30-fold>controls) has been identified. High A4 cows have similar theca molecular phenotypes to women with polycystic ovary syndrome (PCOS) including: irregular estrous cycles, increased inflammation, and reduced fertility. Because PCOS is identified at puberty, the manner heifers attain puberty was investigated.   Heifers were classified using progesterone (P4) concentrations ≥ 1ng/ml to initiate puberty and continued cyclicity: 1) Typical- 378±2 day of age (DOA) (n=279); 2) Early- 317±4 DOA (n=143); 3) Start-Stop- P4 ≥1ng/ml at 265±4 but discontinued cyclicity (n=91); 4) Non-Cycling heifers- no occurrence of P4 ≥ 1ng/ml during sampling period (n=98). Start-Stop and Non-Cycling heifers also had excess A4 secretion from ovarian cortex cultures similar to High A4 cows.   Determined that weaning weight (p=0.017) was reduced in Start-Stop heifers and yearling weights (p=0.0074) was reduced in Start-Stop and Non-Cycling compared to Typical/Early heifers. Non-cycling heifers had the greatest Antral Follicle Counts (p<0.0001) with reduced uterine horn diameter (p=0.0053) compared to Typical (Control, p<0.0001) heifers.   Discovered that there was a lower proportion of Start-Stop and Non-Cycling heifers with a reproductive tract score of 5 compared with Typical/Early heifers, and less response to prostaglandin synchronization causing reduced calving in the first 21 days of the breeding season in the Non-cycling group.   Interestingly, heifers in the Start-Stop group that do not regain cyclicity have similar growth and reproductive traits as the Non-cycling heifers. Taken together, reduced growth and maturation observed in the Start-Stop and Non-Cycling heifers is initiated early in development and continues to adversely affect timing of reproductive maturity and longevity in these heifers.     Determined that there was a tendency (P ≤ 0.09) for increased 17-OH progesterone and 11-deoxycortisol and fibrosis in Non-Cycling heifers. Cytokines CCL5, decorin, and IL-10 were also greater (P ≤ 0.05) in Unstimulated Non-cycling heifers with a tendency (P ≤ 0.09) for increased IFNβ, IL-1β, IL-17, CCL4, and IL-36Ra.   Determined that FSH-stimulation in Non-cycling heifers tended to reduce fibrosis, decorin, IFNβ, IL-1β, and CCL5 (P ≤ 0.03) to the level of the Unstimulated Typical (Control) heifers. In contrast, TNFα was increased (P = 0.05) in FSH-stimulated Non-Cycling heifers compared to other groups. Taken together, these results indicate that FSH-stimulation may restore the ovarian microenvironment of Non-cycling (High A4) heifers and reduce inflammation which may be a cause of follicle arrest.   We conducted a GWAS using 150,000 SNP markers (Geneseek GenomicProfiler Bovine 150K) genotyped in 200 individuals. As a result, we have identified 70 genes within 25 kb of genetic variants significantly (p<0.01) associated with the High A4 phenotype. Additionally, whole-genome sequence of 15 individuals (8 High A4, 7 Control) resulted in the identification of 4,600 variants with predicted high impact (via SNPEff). A total of ~17 million variants were observed in the sample.   High impact variants were identified in seven of the 70 genes identified in the initial GWAS.  We have assayed expression of CARNS1, CCR6, GPR31, ALKBH6, CLIP3, SREBP1a and SREBP1c via ddPCR in ovarian tissue, ovarian cortex, theca, and granulosa cells, in High A4 compared to controls cows.   In High A4 ovarian cortex, SREBP1c and CLIP3 were significantly upregulated compared to controls. There was a tendency for CARNS1 and CCR6 to be downregulated in High A4 granulosa cells. Three of the genes have a role with inflammatory regulation and one has a role in β-alanine metabolism of the High A4 phenotype. Understanding genetic variants can provide a potential tool for determining High A4 cows and removal from the herd.

Publications

Peer-reviewed journal articles reporting research from this project   Abuajamieh, M, Kvidera, SK, Mayorga, EJ, Kaiser, A, Lei, S, Seibert, JT, Horst, EA, Sanz Fernandez, MV, Ross, JW, Selsby, JT, Keating, AF, Rhoads, RP, Baumgard, LH 2018. The effect of recovery from heat stress on circulating bioenergetics and inflammatory biomarkers. Journal of Animal Science. 96(11):4599-4610.   Horst, EA, Kvidera, SK, Mayorga, EJ, Shouse, CS, Al-Qaisi, M, Dickson, MJ, Ydstie, J, Ramirez Ramirez, HA, Keating, AF, Dickson, DJ, Griswold, KE, Baumgard, LH. 2018. Effect of chromium on bioenergetics and leukocyte dynamics following immunoactivation in lactating Holstein cows. Journal of Dairy Science. 101(6):5515-5530.   Hristov, AN,  Degaetano, AT,  Rotz, CA, Hoberg, E, Skinner, RH, Felix, T, Li, H, Patterson, PH, Roth, G, Hall, M, Ott, TL, Baumgard, LH, Staniar, W, Hulet, RM, Dell, CJ, Brito, AF, Hollinger, DY. 2018. Climate change effects on livestock in the Northeast US and strategies for adaptation. Climatic Change 146 (1-2); pgs 33-45 doi.org/10.1007/s10584-017-2023-z   Hughes CK, Maalouf SW Liu W-S, Pate JL. 2019. Molecular profiling demonstrates modulation of immune cell function and matrix remodeling during luteal rescue. Biology of Reproduction, 2019, 0(0), 1–16 doi:10.1093/biolre/ioz037 Research Article Advance Access Publication Date: 27 March 2019   Hughes CK, Pate JL. 2019. Luteolysis and the Corpus Luteum of Pregnancy. In: The Ovary 3rd Edition, PCK Leung and EY Adashi, eds., Academic Press, Elsevier, pp. 270-295.  ISBN 978-0-12-813209-8   Ott, TL. 2019. Symposium Review: Immunological detection of the ruminant conceptus. Journal of Dairy Science 104(4): pgs 3766-3777 doi.org/10.3168/jds.2018-15668    Pate JL, Hughes CK. 2018. Applications of large-scale molecular profiling techniques to the study of the corpus luteum. Anim. Reprod. 15:Suppl. 1, p. 791-804. DOI: 10.21451/1984-3143-AR2018-0038   Pate, JL. 2018. Luteolysis. In M. K. Skinner (Ed.), Encyclopedia of Reproduction. vol. 2, pp. 106–113. Academic Press: Elsevier. <a href="http://dx.doi.org/10.1016/B978-0-12-801238-3.64397-0">http://dx.doi.org/10.1016/B978-0-12-801238-3.64397-0</a>   Silveira, PAS, WR Butler, SE LaCount, TR Overton, CC Barros, and A Schneider. 2019. Polymorphisms in the anti-oxidant paraoxonase-1 (PON1) gene associated with fertility of postpartum dairy cows. Theriogenology 125:302-309.   Bedford A, Beckett L, Hardin K, Dias NW, Davis T, Mercadante VR, Ealy AD and White RR. 2018. Propionate affects insulin signaling and progesterone profiles in dairy heifers. Sci. Rep. 8:17629.   Bonometti S, Menarim BC, Reinholt BM, Ealy AD and Johnson SE. 2019. Growth factor modulation of equine trophoblast mitosis and prostaglandin gene expression. J. Anim. Sci. 97:865-873.    Ealy AD, Wooldridge LK and McCoski SR. 2019. Post-transfer consequences of in vitro-produced embryos in cattle. J. Anim. Sci. ePub 04/10/2019.    Fontes PL, Oosthuizen N, Ciriaco FM, Sanford CD, Canal LB, Pohler KG, Henry DD, Mercadante VR, Timlin CL, Ealy AD, Johnson SE, DiLorenzo N and Lamb GC. 2019. Impact of fetal vs. maternal contributions of Bos indicus and Bos taurus genetics on embryonic and fetal development. J. Anim. Sci. 97:1645-1655.   McCoski SR, Poole RK, Vailes MT and Ealy AD. 2018. Maternal obesity alters the expression of embryonic regulatory transcripts in the preimplantation ovine conceptus. Repro. Biol. 18:198-201.   McCoski SR, Vailes MT, Owens, CE Cockrum RR and Ealy AD. 2018. Exposure to maternal obesity alters gene expression in the preimplantation ovine conceptus. BMC Genomics 19:737.   Vailes MT, McCoski SR, Wooldridge LK, Reese ST, Pohler KG, Roper DA, Mercadante VR and Ealy AD. 2019. Post-transfer outcomes in cultured bovine embryos supplemented with epidermal growth factor, fibroblast growth factor 2, and insulin-like growth factor 1. Theriogenology 124:1-8.   Wooldridge LK and Ealy AD. 2019. Interleukin-6 increases inner cell mass numbers in bovine embryos. BMC Dev. Biol. 19:2.   Zhang Z, Zhao LD, Johnson SE, Rhoads ML, Jiang H and Rhoads RP. 2019. Oxytocin is involved in steroid hormone-stimulated bovine satellite cell proliferation and differentiation in vitro. Domest. Anim. Endocrinol. 66:1-13.   Carvalho PD, Santos VG, Giordano JO, Wiltbank MC, Fricke PM. 2018. Development of fertility programs to achieve high 21-day pregnancy rates in high-producing dairy cows. Theriogenology 2018; 114:165-172.  10.1016/j.theriogenology. 03.037   Garcia-Guerra A, Canavessi AMO, Monteiro PLJ, Jr., Mezera MA, Sartori R, Kirkpatrick BW, Wiltbank MC. 2018. Trio, a novel bovine high fecundity allele: III. Acquisition of dominance and ovulatory capacity at a smaller follicle size. Biol Reprod 98:350-365.  10.1093/biolre/iox157   Garcia-Guerra A, Kamalludin MH, Kirkpatrick BW, Wiltbank MC. 2018. Trio a novel bovine high-fecundity allele: II. Hormonal profile and follicular dynamics underlying the high ovulation rate. Biol Reprod 98:335-349.  10.1093/biolre/iox156   Garcia-Guerra A, Wiltbank MC, Battista SE, Kirkpatrick BW, Sartori R. 2018. Mechanisms regulating follicle selection in ruminants: lessons learned from multiple ovulation models. Animal Reproduction 15:660-679.    Kamalludin MH, Garcia-Guerra A, Wiltbank MC, Kirkpatrick BW. 2018. Proteomic analysis of follicular fluid in carriers and non-carriers of the Trio allele for high ovulation rate in cattle. Reprod Fertil Dev  10.1071/RD17252   Kamalludin MH, Garcia-Guerra A, Wiltbank MC, Kirkpatrick BW. 2018. Trio, a novel high fecundity allele: I. Transcriptome analysis of granulosa cells from carriers and noncarriers of a major gene for bovine ovulation rate. Biol Reprod 98:323-334.  10.1093/biolre/iox133   Melo LF, Monteiro PLJ, Jr., Nascimento AB, Drum JN, Spies C, Prata AB, Wiltbank MC, Sartori R. 2018. Follicular dynamics, circulating progesterone, and fertility in Holstein cows synchronized with reused intravaginal progesterone implants that were sanitized by autoclave or chemical disinfection. J Dairy Sci 101:3554-3567.  10.3168/jds.2017-13570   Melo LF, Monteiro PLJ, Jr., Oliveira LH, Guardieiro MM, Drum JN, Wiltbank MC, Sartori R. 2018. Circulating progesterone concentrations in nonlactating Holstein cows during reuse of intravaginal progesterone implants sanitized by autoclave or chemical disinfection. J Dairy Sci 101:3537-3544.  10.3168/jds.2017-13569   Ochoa JC, Penagaricano F, Baez GM, Melo LF, Motta JCL, Garcia-Guerra A, Meidan R, Pinheiro Ferreira JC, Sartori R, Wiltbank MC. 2018. Mechanisms for rescue of corpus luteum during pregnancy: gene expression in bovine corpus luteum following intrauterine pulses of prostaglandins E1 and F2alpha. Biol Reprod 98:465-479.  10.1093/biolre/iox183   Silva EP, Wiltbank MC, Machado AB, Gambin LS, Dias MM, Chaiben MFC, Bernardi ML, Borges JBS. 2018. Optimizing timed AI protocols for Angus beef heifers: Comparison of induction of synchronized ovulation with estradiol cypionate or GnRH. Theriogenology 121:7-12.  10.1016/j.theriogenology.2018.07.019   Vasconcelos JLM, Pereira MHC, Wiltbank MC, Guida TG, Lopes Jr. FR, Sanches Jr. CP, Barbosa LFSP, Costa Jr. WM, Munhoz AK. 2018. Evolution of fixed-time AI in Brazil. Animal Reproduction 15: 940-951.    Wiltbank MC, Mezera MA, Toledo MZ, Drum JN, Baez GM, Garcia-Guerra A, Sartori R. 2018. Physiological mechanisms involved in maintaining the corpus luteum during the first two months of pregnancy. Animal Reproduction 15:805-821.   Tenley SC, Spuri-Gomes R, Rosasco SL, Northrop EJ, Rich J, McNeel AK, Summers AF, Miles JR, Chase CC, Lents CA, Perry GA, Wood JR, Cupp AS, Cushman RA. 2018. Maternal Age Influences the Number of Primordial Follicles in the Ovaries of Yearling Angus Heifers, Animal Reproduction Science 200:105-112   McFee RM, Cupp AS, and Wood JR. 2018. Use of case-based or hands-on lab exercises with physiology lectures improves knowledge retention but veterinary medicine students prefer case-based activities, Adv Physiology Education, 42(2):182-191   Wood JR, and Cupp AS. 2018. Aromatization. In M. K. Skinner (Ed.), Encyclopedia of Reproduction. vol. 2, pp. 195–201. Academic Press: Elsevier. <a href="http://dx.doi.org/10.1016/B978-0-12-801238-3.64642-1">http://dx.doi.org/10.1016/B978-0-12-801238-3.64642-1</a>   Ganesan, S, Dickson, MJ, Keating, AF. 2018. Pesticides. In M. K. Skinner (Ed.), Encyclopedia of Reproduction. vol. 2, pp. 724–730. Academic Press: Elsevier. Clark, KL, Ganesan, S, Keating, AF. 2018. Impact of Toxicant Exposures on Ovarian Gap Junctions. Reproductive Toxicology. 81:140-146. Bidne, KL, Dickson, MJ, Ross, JW, Baumgard, LH, Keating, AF. 2018. Disruption of female reproductive function by endotoxins. Reproduction. 155(4):R169-R181.     Conference papers and abstracts/presentations Clark, KL, Talton, O, Ganesan, A, Schulz, LC, Keating AF. 2018. Lean maternal gestational diabetes mellitus impacts the ovarian proteome basally and in response to a stressor in later life. Society for the Study of Reproduction annual meeting, New Orleans.   Parrish, JJ, Keating, AF, Pate, JL, Pohler, KG, Wiltbank, MC, Ealy, AD. 2018. Changing paradigm in graduate education: Collaborative on-line approach in Reproduction. American Society of Animal Science annual meeting.   Piet S, Walker S, Tsang PCW.  2019. Expression and Regulation of Cysteine Rich 61-Connective Tissue Growth Factor-Nephroblastoma Overexpressed (CCN1) in Ovarian Adenocarcinoma (OVCAR8) Cells. 52nd Annual Meeting of the Society for the Study of Reproduction, San Jose, CA.   Wiltbank MC, Carvalho PD, Barletta RV, Fricke PM, Shaver RD. 2018. Relationships Among Changes in Body Condition Score and Reproductive Efficiency in Lactating Dairy Cows. 27th Tri-State Dairy Nutrition Conference 2018:121-138.    Goulet M, Donahue J, Tsang PCW. 2019. Regulation of the Angiogenic Inducer, Cysteine Rich 61-Connective Tissue Growth Factor-Nephroblastoma Overexpressed 1 (CCN1), in Bovine Luteal Cells. 52nd Annual Meeting of the Society for the Study of Reproduction, San Jose, CA.   Lundberg, Jaskiewicz NJ, Townson DH. 2019. Stimulatory effects of Transforming Growth Factor-alpha in bovine granulosa cells of small antral follicles. Society for the Study of Reproduction Annual Meeting   Arbeithuber B, Stoler N, Higgins B, Hester JM, Diaz F, Nekrutenko A, Makova KD. 2018. Rare mitochondrial DNA variant analysis in single oocytes using duplex sequencing. 31st annual meeting of The Biology of Genomes. Cold Springs Harbor Laboratory.  Cold Springs Harbor, NY. May 8-12.   Baddela VS, Hughes CK and Pate JL. 2018. Let7 miRNA are abundant in the bovine corpus luteum and regulate cellular metabolic functions Society for the Study of Reproduction, New Orleans, LA   Butler WR, Schneider A, Silveira PAS, Townson DH, Tsang PCW, Dailey RA, Ott TL and Pate JL. 2018. Effects of polymorphisms in GHR, IGF-I and TNFα genes on fertility in lactating dairy cows. Amer Dairy Sci Association   Butler, W.R., Schneider., A., Silveira. P.A.S., Townson, D.H., Tsang, P.C.W., Dailey, R.A., Diaz FJ. 2018. Sufficient Dietary Zinc During the Preconception Period is Essential for Oocyte Competence.  Endocrine Society Annual Meeting, Chicago, IL. March 17-20.   Harper, MT, Oh, J, Melgar, A, Nedelkov, K, Räisänen, S, Chen, X, Martins, CMMR, Young, M, Ott,TL, Kniffen, DM, Fabin, R, Hristov, AN. 2018. Production effects of extruded soybean meal in early lactation cow diets. Journal of Dairy Science, 102 (Suppl 2) Abst: 510.   Hester JM and Diaz FJ. 2018. Growing Oocytes Need Zinc: Zinc Deficiency in the Preantral Ovarian Follicle.  Experimental Biology Meeting, San Diego CA.  *Endocrinology and metabolism section research recognition award.   Hughes CK, Maalouf SW Liu W-S, Pate JL. 2019. Molecular profiling demonstrates modulation of immune cell function and matrix remodeling during luteal rescue. Biology of Reproduction, 2019, 0(0), 1–16 doi:10.1093/biolre/ioz037 Research Article Advance Access Publication Date: 27 March 2019   Hughes CK, Rogus A, Inskeep EK and Pate. 2018. Functional transitions in the corpus luteum are associated with changes in NR5A2 abundance, which regulates luteal progesterone production. Proc. 10th Int Ruminant Reprod Symp, Foz do Iguacu, Brazil   Hughes CK, Wetzel LM, Bosviel R, Newman J and Pate JL. 2018. Luteal metabolomics suggests a role for lipid mediators as regulators of cell death, cell migration, and immune function. Society for the Study of Reproduction, New Orleans, LA   Lupicka M and Pate JL. 2018. Luteal Extracellular Vesicles Carry MicroRNA and Modulate Cytokine Secretion from Immune Cells. Society for the Study of Reproduction, New Orleans, LA   Melgar, A, Harper, MT, Oh, J, Giallongo, F, Fetter, ME, Ott, TL, Duval, S, Hristov, AN. 2018. Effects of 3-nitrooxypropanol on rumen fermentation, lactational performance, and onset of ovarian activity in dairy cows. Journal of Dairy Science, 102 (Suppl 2) Abst: 209.   Ott, TL, Pate, JL. 2018. Effects of polymorphisms in GHR, IGFI and TNFA genes on fertility in lactating dairy cows. Journal of Dairy Science, 102 (Suppl 2) Abst. T207   Ott, TL., 2018. Immune recognition of the periattachment conceptus. Journal of Dairy Science, 102 (Suppl 2) Abst; 102.   Pate JL and Hughes CK. 2018. Applications of large-scale molecular profiling techniques to the study of the corpus luteum. Proc 10th Intl Ruminant Reproduction Symposium, Foz do Iguacu, Brazil Ealy AD. 2019. Early Diagnosis of Pregnancy Loss. American Dairy Science Association Annual Meeting, Cincinnati OH.   Wooldridge LK, Johnson SE and Ealy AD. 2019. Interleukin-6 increases inner cell mass and hypoblast cell numbers in bovine blastocysts. Society for the Study of Reproduction 52nd Annual Meeting, San Jose CA.   Wooldridge LK and Ealy AD. 2019. STAT3 is required for hypoblast lineage development in bovine blastocysts. 2019. Society for Developmental Biology 78th Annual Meeting, Boston MA.   Cushman RA, Tenley SC, Spuri-Gomes R, Rosasco SL, Northrop EJ, Rich JRR, McNeel AK, Summers AF, Miles JR, Chase C, Lents CA, Perry GA, Wood JR, and Cupp AS. 2018. Influence of Maternal Age on Daughter Ovarian Reserve and Reproductive Longevity in Beef Cows. 51st Annual Meeting of the Society for the Study of Reproduction, New Orleans, LA   Neilson MA, Spuri-Gomes R, Romereim SM, Summers AF, Abedal-Majed MA, Tenley SC, Kurz SG, Bergman J, Davis JS, Wood JR, and Cupp AS. 2018. Altered Blood Plasma and Follicular Fluid Lipid Profiles Suggest Possible Discrepancies in Cell Signaling and Metabolism in Cows with Androgen Excess. 51st Annual Meeting of the Society for the Study of Reproduction, New Orleans, LA   Snider AP, Romereim SM, Summers AF, Pohlmeier WE, McFee RM, Spuri-Gomes R, Kurz SG, Davis JS, Wood JR, and Cupp AS. 2018. Exposure to Excess Androgen in the Ovarian Microenvironment of High A4 Cows Results in Altered Function of Granulosa Cells which may Explain Changes in Cyclicity and Response to Male Exposure. 51st Annual Meeting of the Society for the Study of Reproduction, New Orleans, LA   Hart ML, Abedal-Majed MA, Spuri-Gomes R, Snider AP, Kurz SG, Bergman JW, McFee RM, Casey CA, Davis JS, Cushman RA, Wood JR, and Cupp AS. 2018. Cyclicity Phenotype and Ovarian Cortex Androgen Secretion in Androgen Excess Cows are Predictive of Plasma Steroid and Lipids, Liver Enzymes, and Follicular Fluid Cytokines. 51st Annual Meeting of the Society for the Study of Reproduction, New Orleans, LA   Bochantin KA, Summers AF, Pohlmeier WE, Sargent KM, Kurz SG, Romereim SM, Daudu O, McFee RM, Cushman RA, Davis JS, Cupp AS, Wood JR. 2018.  Microarray Analysis Predicts that Differentially Expressed Genes in Theca Cells from Cows with High Intrafollicular Androstenedione are Regulated by ESR1 and VEGFA Signaling and Increased mRNA Stability. 51st Annual Meeting of the Society for the Study of Reproduction, New Orleans, LA   Timme KR, Miller HB, and Wood JR. Signaling of Pro-Inflammatory Cytokine TNFa through NFkB-p65 Increases Gdf9 and Decreases Figla mRNAs in Oocytes of Primary Follicles. 2018. 51st Annual Meeting of the Society for the Study of Reproduction, New Orleans, LA   McCain AR, Rister AL, Wordekemper M, Dodds ED, and Wood JR. 2018. Maternal obesity due to diet or loss of satiety differentially impacts fetal growth and placental efficiency which may be due to differences in excess circulating lipids. 51st Annual Meeting of the Society for the Study of Reproduction, New Orleans, LA   Theses/Dissertations   Hester, JM. The impact of zinc deficiency during oogenesis, follicle assembly, and growth. PhD Thesis in Physiology, Pennsylvania State University. Dec 2018.   Audra Harl. 2018. Factors affecting the quality and function of the bovine periovulatory follicle. (PhD Dissertation).   Sarah McCoski. 2018. Manipulating embryonic development and endometrial function in ruminants. (PhD Dissertation).   Lundberg, Townson DH. 2019 Investigation of Transforming Growth Factor-α and its potential role in promoting ovarian follicular dominance. UVM MS Thesis   Active collaboration within the group   NY, PA, NH, VT and WV:  Samples collected to investigate associations between fertility outcomes and SNP in candidate genes from dairy cows.   IA and UK: Samples shared to determine the effect of LPS treatment on ovarian inflammation.   PA and WV: Samples shared for analysis of molecular regulators of luteal function.   VT and NY: Samples assayed (NY) for analysis of progesterone concentrations.     In addition to Hatch Multistate Funds, these studies were supported by:   Iowa Pork Producers Association                                                                                                  National Pork Board   ISU Nutritional Sciences Council Martin Fund       National Institute of Environmental Health Science - Agriculture and Food Research Initiative   National Institutes of Health   C. Lee Rumberger and Family Endowment   John L. Pratt Animal Nutrition Program   Virginia Agricultural Council Research Funding   University of Nebraska Foundation Funds

Impact Statements

Low pregnancy rates in beef and dairy cows result in increased costs to producers associated with rebreeding or culling animals, and recouping heifer development costs. Reduced fertility is also a major cause of reduced milk (dairy) and meat (beef) production which impacts the food supply. The focus of the NE1727 multistate project “Influence of ovary, uterus, and embryo on pregnancy success in ruminants”, is to understand how altered ovarian function, impaired oocyte quality, and disruptions of conceptus-uterine interactions contribute to infertility, and devise management strategies that will overcome these factors in order to optimize the chances that animals successfully achieve a pregnancy. To achieve this goal, the following three objectives will be performed: (1) Determine the impact of altered ovarian function on reproductive performance; (2) Identify alterations in embryo development and uterine and CL function associated with declining pregnancy establishment; and (3) Identify changes in genetics and reproductive management that lead to improved pregnancy rates. As investigators in NE1727, we will use our collective expertise to understand the mechanisms by which internal and external forces modify these important physiological processes and to use this information about core biological processes to rationally develop new and innovative tools to increase cyclicity and breeding efficiency, and reduce early embryonic failures, thereby improving reproductive efficiency of both dairy and beef cattle operations in the United States.

Date of Annual Report: 08/13/2020

Report Information

Annual Meeting Dates: 05/19/2020 - 05/20/2020
Period the Report Covers: 10/01/2019 - 09/30/2020

Participants

Michelle Rhoads
Phil Bridges
Dave Townson
Vimal Selvaraj
Joanne Fortune
Milo Wiltbank
Aileen Keating
Alan Ealy
Ben Crites
Cecily Bishop
Dan Mathew
Erdogan Memili
Alvaro Garcia Guerra
Gary Thompson
Gustavo Grillo
Heather Baldwin
Jessica Cristina
Jaelyn Current
Jaspreet Rishi
Jen Wood
Jianbo Yao
Joy Pate
Julio Giordano
Keith Inskeep
Mark Mirando
Mingxiang Zhang
Pablo Visconti
Paul Tsang
Pedro Monteiro
Rafael Fissore
Rafael Reis Domingues
Bob Dailey
Rodolfo Cardosa
Sarah Carr
Victor Gomez Leon
JP Andrade
Ron Butler
Yi Athena Ren
Troy Ott
Alicia Arneson
Jessica Motta
Estefenia Gonzalez Alvares
Dan Poole
Abby Maucieri
Adam Beard
Crystal Roach
Francisco Diaz
Sarah Piet

Brief Summary of Minutes

Minutes Attachment:

NE1727 Minutes. 2020. Virtual.docx

Accomplishments

The following have been accomplished: The bovine high fecundity allele, Trio, results in overexpression of SMAD6 and a 3-fold increase in ovulation rate. Once activated, Trio carrier follicles, have reduced progression through the primary stage, hence the larger oocyte and greater number. The effect of unilateral ovariectomy (ULO, D0) on antral follicle count (AFC), anti-Müllerian Hormone (AMH) and ovulation rate in carriers and non-carriers of the Trio allele (n = 9/group) was evaluated. Unique observations were: 1) ULO decreased AFC and AMH to ~50%, 2) AFC eventually returned to pre-ULO values by 2 (Trio carriers) to 4 (non-carriers) months, and 3) ovarian compensation, based on AMH, occurred in Trio carriers (~8 months) but not in non-carriers by conclusion of the study (D230). Induced embryo demise results in luteolysis, through ill-defined mechanisms, and is associated with timing of conceptus expulsion. The induction of embryonic demise may be a valuable model to investigate the mechanisms regulating luteolysis or CL maintenance during the second month of gestation. Found evidence of presence of a continuous mechanism for CL maintenance beyond the classical maternal recognition period in cattle.   Previous research has shown that induction of an accessory CL by administration of GnRH on d5 increases circulating progesterone and reduces pregnancy loss between d33 and d60 in heifers receiving IVP embryos.  Treatment with GnRH caused ovulation, formation of an accessory CL, however, there was no effect of timing of treatment on pregnancies per ET nor pregnancy loss. Found that the removal of the initial GnRH from a 5-day FTET protocol resulted in a slight but significant reduction in the utilization rate, however, there was no detrimental effect on fertility. As a result, the overall cost of the FTET program can be reduced by eliminating the need for the initial GnRH treatment without compromising fertility. Determined that pregnancies per artificial insemination (P/AI_ was similar (P=0.35) between heifers receiving a Single (43.6%; 65/149) or a Double (38.7%; 58/150) dose of GnRH at initiation of the protocol. Similarly, in cows neither estrous expression [57.1% (313/548) vs 56.0% (310/553)] nor P/AI [49.5% (271/548) vs 49.6% (274/553)] were different (P>0.60) between Single and Double GnRH treated, respectively. In heifers, increasing the dose of GnRH, resulted in a greater (P=0.04) ovulation rate in heifers in the Double dose group (40.9%; 36/88) than those in the Single dose group (26.1%; 23/88), while in cows ovulation to the initial GnRH was not different (P=0.35) between cows treated with a Single or Double dose of GnRH [66.5% (189/284) vs 68.4% (195/285)].  Finally, heifers that did ovulate after the initial GnRH had greater (P=0.04) P/AI (52.5%) than heifers that did not ovulate (40.2%), while P/AI did not differ between cows that did (54.9%) or did not (57.0%) ovulate to the initial GnRH.   Found that treatment with GnRH at the time of AI is a suitable strategy to improve P/AI in heifers that fail to exhibit estrous after an estradiol-based FTAI protocol.  Likewise, cows that did not exhibit estrous and received GnRH at the time of AI had greater (P<0.01) P/AI than those that did not exhibited estrous and remain untreated (43.9% vs 32.5%). Pregnancy loss was not different between any of the groups (P>0.10). Morphometric characteristics of the female reproductive tract are primarily determined during prenatal development and may have the potential when assessed to aid in the prediction of fertility in beef cattle.  Marked variation of morphometric characteristics of the reproductive tract among individuals was observed, however, total ovarian area was the only factor associated with pregnancy outcomes to FTAI.   Early pregnancy detection has the potential to improve reproductive efficiency of embryo transfer (ET) programs by identifying non-pregnant animals and allowing their prompt reutilization.  Sensitivity was lowest (89.8%) for uterine endometrial thickness (UET) and was different (P<0.01) from the other three tests. Specificity was greatest (P<0.01) for serum P4 (67%), lowest for UET (33.9%), and intermediate for CL volume (55.4%) and Color Doppler (58.9%). Serum P4, CL volume and CL Color Doppler showed excellent NPV (>97%) indicating that each of these values accurately predicted non-pregnant heifers. However, PPV was moderate (70 to 75%) indicating the potential for false positive results either due to pregnancy loss between d 21 and d 28 or, alternatively, inaccuracy of the method. Found that pregnancy diagnosis by color Doppler ultrasonography at d 22 after FTAI coupled with the use of a CIDR may be an effective strategy to identify non-pregnant heifers and attempt their prompt re-synchronization.  Color Doppler ultrasonography had less Specificity (59.1% compared with 90.2%) and PPV (75% compared with 89.3%) in heifers than cows. In conclusion, color Doppler ultrasonography settings that were evaluated did not affect the diagnostic performance for pregnancy diagnosis at d21. Ewes administered PG-600 near the end of a CIDR-synchronization protocol displayed higher progesterone levels than controls despite similar number of CL in their subsequent estrous cycle. This was accompanied by a downregulation of follicle stimulating hormone receptors in the glandular epithelium and high expression of MUC-1 in luminal epithelium in early diestrus (7 days post-treatment). This is postulated to explain the subfertility of ewes synchronized with this regimen of CIDR/PG-600. Culture systems to support development of isolated follicles from preantral to antral stages have many applications, ranging from basic ovarian biology to assisted reproductive techniques for women and valuable mammalian species. Unfortunately, a standard, “best practices” protocol does not exist for many species of interest. A 3D, matrix-free low attachment culture system was identified which supported growth of multilayer bovine follicles to small antral stage. Addition of epidermal growth factor (EGF) appears to support survival, antrum formation and growth of follicles to greater sizes than previously reported with matrix-based methods (~2 mm vs 0.5 mm). Addition of EGF+Kit Ligand improved growth of some follicles, while addition of EGF+LIF showed modest improvement. However, follicles have yet to reach sizes associated with oocytes capable of in vitro maturation (3-6 mm).    Bull fertility is the ability of the sperm to fertilize and activate the egg and support embryo development. Sperm freezability or cryopreservability if the post-thaw viability of sperm following freezing and thawing. Research shows that there are significant differences in fertility and sperm freezability among breeding bulls.  Outcomes of the research enhance fundamental science and technology on sperm production, function, and cryopreservation-discerning bull fertility and sperm freezability. Treatment of bull sperm with ionophore A23187 in standard TALP medium rapidly immobilized bull sperm. Important, when A23187 was washed away 10 minutes later, bull sperm rapidly regained general motility, and these sperm displayed parameters consistent with hyperactive motility such as decrease linearity, increased VCL (curvilinear velocity) and ALH (lateral head displacement). When ionophore A23187-treated bull sperm were functionally tested for in vitro fertilization rates, ionophore treated sperm performed better than control sperm, but not better than heparin-treated sperm. However, the combined treatment of ionophore A23187+ heparin, produced significantly better results than either of the treatments alone, especially when the outcomes were evaluated as total number of blastocysts and total number of hatched blastocysts. “Sperm Energy Recovery after starvation” (SER) treated bull sperm, just as mouse sperm treated by the same procedure, responded to the restoration of energy with better motility patterns, and increased rates of pre-implantation of embryo development. “Sperm Energy Recovery after starvation” (SER)-treated bull sperm also resulted in better fertilization rates following ICSI, greater percentages of sperm induced oscillations, the rates of blastocysts development changed dramatically. “Sperm Energy Recovery after starvation” (SER)-treated sperm seem to undergo differential re-localization of sperm PLCzeta, the sperm molecule responsible for the initiation of oscillations at fertilization in all mammals. Gilts were treated as follows to determine an additive impact of thermal stress on the ovarian response to chemical exposure: 1) Thermal Neutral ad libitum feed intake, 2) Thermal Neutral ad libitum feed intake plus Zearalenone, 3) Thermal Neutral feed intake paired to heat stressed gilts (group 5 and 6), 4) Thermal Neutral feed intake paired to heat stressed gilts plus Zearalenone, 5) Heat Stressed ad libitum feed intake, 6) Heat Stressed ad libitum feed intake plus Zearalenone. Tissues have been collected and molecular analysis is underway (Goal #1). The impact of thermal stress on the ovarian prolactin signaling pathway has been evaluated in gilts who were heat stressed during the follicular phase of their estrous cycle. This is potentially an important mode of heat stress-altered ovarian function since prolactin is elevated in heat stressed production animals (Goal #1). Using LC-MS/MS, we have completed proteomic analysis of ovaries from mice who were exposed to perfluorooctonic acid (PFOA), or Zearalenone (ZEA) or Glyphosate (GLY). We have collected phenotypic data from each of these separate experiments. In addition, both lean and obese mice were included in each study and the impact of altered metabolic status has been determined (Goal #1). Completed immunological staining to investigate the location and abundance of ovarian connexin 37 and 43 to understand communication between somatic cells and the female gamete.  This staining was performed in ovarian sections from lean or obese mice who were exposed to vehicle control or PFOA (Goal #2). Utilized DNA repair protein deficient mice as well as obese mice to investigate whether obesity alters the capacity of the ovary to discard damaged follicles (Goal #1). Fecal DNA, which largely represents bacterial DNA, was isolated and whole genome shotgun sequencing performed.  A total of 36 samples were sequenced as follows: Control/unstimulated (), High A4/unstimulated (), Control/FSH stimulated, High A4/FSH stimulated (). The analyses will determine representation of microbe families in each experimental group and similarities or differences in the presence of functional genes between experimental groups. Blood plasma was assayed to detect subclinical inflammation. A cytokine array panel was interrogated and the upper and lower quartile of cows based on their TNFα concentrations were identified.  TNFα concentration was also positively correlated to several other pro- and anti-inflammatory cytokines indicating a systemic inflammatory response. Follicular fluid and theca cells from Large (> 7mm) and small (< 5mm) follicles was collected at ovariectomy. Samples were classified based on control vs. high TNFa (circulating).  Androstenedione but not progesterone was decreased in follicular fluid in both large and small follicles. We performed preliminary BODIPY staining in theca sections. It increases as theca undergo luteinization Determined that TNFa was higher in cows that were acyclic prior to their first breeding (delayed puberty). This suggests that the subclinical inflammatory response may be chronic and this enhanced response to stress could be programmed prior to puberty. Using a mouse model, we have collected oocytes and cumulus granulosa cells from conventional and germ-free mice fed a control or high fat/high sugar diet. Analyses will be carried out in 2020. Expression of ZIM2 in bovine oocytes and early embryos was characterized. A potential ZIM2 binding element (ACGTTACCCT) was identified and the interaction between ZIM2 protein and KAP1 (co-repressor) was confirmed. Differential expression of bovine long noncoding RNAs in oocytes derived from small and large follicles was identified. Expression of agouti signaling protein (ASIP) gene in ovarian tissues and early embryos was characterized.  Chronic exposure to ergot alkaloids decreases the utero-ovarian blood vessels, thus reducing blood flow to the ovary. Chronic exposure to ergot alkaloids in addition to heat stress disrupts folliculogenesis, thus hindering the transition from recruited to preovulatory follicles. Exposure to ergot alkaloids during the second trimester of gestation does not hinder subsequent heifer growth and reproductive performance. No differences in primordial, recruited, selected or preovulatory follicles were observed. Chronic exposure to ergot alkaloids reduces corpus luteum area and progesterone production compared to control animals. Presence of a functional CL at the start of a FTAI protocol improves pregnancy success in postpartum beef cows, thus incorporating a presynchronization program prior to FTAI will increase pregnancy rates. The slick hair mutation, found in Senepol cattle, appears to aid in offsetting the physiological symptoms associated with fescue toxicosis and helps to improve reproductive performance. Determined the effects of calcium ionophore (0.1, 0.5 and 1 uM) and ethylene glycol tetraacetic acid (EGTA; 2 mM) on CCN1 expression in ovarian adenocarcinoma (OVCAR8) cells. Determined the effects of calcium ionophore (0.1, 0.5 and 1 uM) on CCN1 expression in OVCAR8 cells treated with signal transducer and activator of transcription 3 (STAT3) small interfering ribonucleic acid (siRNA). Determined the effects of calcium ionophore on cyclic adenosine monophosphate (cAMP) response element binding protein (CREB1) expression in OVCAR8 cells. Determined the effects of CREB1 siRNA on CCN1 expression in OVCAR8 cells. Performed a pilot experiment to determine the effects of insulin-like growth factor 1 (IGF1; 50, 100 ng/mL) and phorbol 12-myristate13-acetate (PMA; 1, 10 nM) on CCN1 and vascular endothelial growth factor (VEGFA) expression in granulosa cells obtained from large, dominant bovine follicles. O-GlcNAcylation is readily evident in bovine granulosa cells (GCs) of both small and large follicles. The GCs of small follicles exhibit greater expression of O-GlcNAcylation than large follicles.  Conversely, the follicular fluid of small follicles contains less glucose and greater lactate than large follicles. Exogenous inhibition of O-GlcNAcylation impairs proliferation of GC in vitro. Determined the most abundant miRNA in extracellular vesicles released from luteal cells, and changes in luteal-resident T cell proteins during luteolysis. The switch in the most abundant miRNA in the CL between days 4 and 6 are those that regulate luteal cell calcium homeostasis and susceptibility to apoptosis, clearly implicating miRNA in acquisition of luteolytic capacity. Profiled temporal, molecular changes in the CL during early pregnancy that provide clues about pathways that are important for continued luteal function.  Developed microfluidic device for high content screening of compounds that block cumulus expansoin, oocyte maturaiton or fertilization. Screened a FDA library of drugs (Prestwick) and found potentially novel pathways involved in cumulus expansion. Determined early pregnancy was accompanied by an increase monocytes in peripheral blood as well as mediators of immune tolerance in dairy heifers. Determined that supplemental omega-3 fatty acids fed to transition dairy cows on a commercial farm improved conception rates in primiparous heifers and milk yield in multiparous dairy cows. The SNP in TNFα had significant effects on fertility; Combining the effects of SNPs in TNFα + IGFI, cows carrying more favorable alleles (AA for each gene) had fewer DIM to pregnancy than cows having BB genotype for both (AAAA=112±8, BBBB=126±10; Linear effect P=0.019). SNPs in PON1 promoter at positions -221 and -392 were significantly associated with both plasma PON1 activity and a reduced calving to conception interval (P<0.05) with no significant effect on calving to first ovulation interval. These SNPs provide novel genetic markers for improved fertility in future studies in dairy cows. Preliminary results confirm significant effects on fertility in lactating dairy cows of a SNP in COQ9.  Identification and level of expression of a family of glycan binding proteins (galectins; LGALS1, 3, 4, 7, 7b, 8, 9, 13L and 15) considered essential for pregnancy establishment in mammals, was characterized for both cattle and sheep conceptuses during the time of early embryonic loss. Galectin-1 (LGALS1) promotes endometrial expression of genes considered essential for pregnancy establishment in cattle and other species including FOXP3, ISG15, CXCR4 and IL6. Bovine endometrial epithelial and fibroblast specific expression of interferon stimulated genes (ISGs), factors considered essential for pregnancy establishment in ruminants, was characterized during the time of maternal recognition of pregnancy and early embryonic loss in cattle. Explored ways to develop induced pluripotent stem cells from cattle and identify biomarkers in bovine serum that can be used in the diagnosis of pregnancy and pregnancy failures. Transvaginal oocyte aspiration of medium-sized follicles every third day for 30 days does not disrupt ovarian follicular development of early lactation dairy cattle.  Likewise, collection of uterine biopsies alongside every other transvaginal oocyte aspiration does not affect cyclicity. The cytokine, interleukin-6, serves as an embryokine in bovine embryos during embryo culture. Its primary actions are to 1) enable development of embryos cultured individually, 2) improve development of the inner cell mass (ICM) through a JAK-dependent mechanism, and 3) promote primitive endoderm development through a JAK-dependent mechanism.   Selective medium supplements can improve ICM cell numbers during in vitro bovine embryo culture. Modifications in early fetal development are observed after transfer of in vitro-produced bovine embryos with interleukin-6. These changes appear to be positive as fetus size more closely resembles fetuses from AI-generated pregnancies rather than fetuses from non-treated in vitro-produced embryos.  We are understanding the mechanisms involved in selection of a single dominant follicle in cattle. This will be key for the rational design of new reproductive management strategies and reproductive biotechnologies. We have discovered the mechanism that produces increased ovulation rate in carriers of the Trio allele.  This mechanism may also underlie double ovulation in dairy cattle and in other species.  This should allow us to manipulate the ovulation rate in cattle and perhaps other species.  We now have clearly demonstrated that the corpus luteum is maintained during early pregnancy by suppression of pulses of prostaglandin F2a by interferon-tau, based on our measurements of PGFM surges. We have also discovered that there is a second period of corpus luteum maintenance after Day 25 when interferon-tau secretion ends and PGF pulses are initiated. The resolution of the mechanisms that allow maintenance of the corpus luteum during this second period will be key for reducing pregnancy loss in cattle. We found that increasing circulating progesterone can reduce pregnancy loss in recipients of in-vitro produced embryos. This provides an easy method to reduce pregnancy loss by treatment with either GnRH or hCG.

Publications

Peer-reviewed journal articles reporting research from this project Abedal-Majed MA, Kurz SG, Springman SA, McNeel AK, Freetly HC, Largen V, Magamage M, Sargent KM, Wood JR, Cushman RA, Cupp AS (2019) Vascular endothelial growth factor A isoforms modulate follicle development independent of diet in peri-pubertal heifers through diverse signal transduction pathways. Biol Reprod 102: 680-692, PMID: 31723977  Akin, E.E., Johnson, A.K., Ross, J.W., Millman, S.T., Jass, C.D., Stinn, J.P., Stalder, K.J. 2019. Modified wean-to-finish mat as an alternative handling tool for moving grow-finish pig cadavers: A pilot study. Journal of Swine Health and Production 27:278-283. Aldrigde EM, Fearon JE, Haynes PB, Miller HM, Sanford KY, Scott RR, Anglin WW, Blolack LS, Burkes BL, Cohn-White OL, Franks BR, Giles HM, Greene AL, Hanby RD, Holliman AG, Kirby JM, Klein AU, Lehman CA, Lloyd GJ, Lore CT, McMurray TB, Moody ZV, Palmer BN, Pansano LV, Pickle RM, Schaeffer LM, Sidl JR, Smith JD, Stepp HF, Satrio FA, Kutchy NA, Dechert E, Rutherford C, Brown K, Purwantara B, Memili E. (2019) Solutions for grand challenges in goat and sheep production. BIOTROPIA. 26(1): 56-64. Basavaraja, R., S. T. Madusanka, J. N. Drum, K. Shrestha, S. Farberov, M. C. Wiltbank, R. Sartori, and R. Meidan. 2019. Interferon-Tau Exerts Direct Prosurvival and Antiapoptotic Actions in Luteinized Bovine Granulosa Cells. Sci Rep 9:14682. Bidne, K.L., Romoser, M.R., Ross, J.W., Baumgard, L.H., Keating, A.F. 2019. Heat stress during the luteal phase decreases luteal size but does not impact circulating progesterone in gilts. Journal of Animal Science. 97(10):4314-4322. Boettcher, A.N., Kiupel, M., Adur, M.K., Cocco, E., Santin, A.D., Bellone, S., Charley, S.E., Blanco-Fernandez, B., Risinger, J.I., Ross, J.W., Tuggle, C.K., Shapiro, E.M. 2019. Human Ovarian Cancer Tumor Formation in Severe Combined Immunodeficient (SCID) Pigs.  Front Oncol. Bonometti S, Menarim BC, Reinholt BM, Ealy AD and Johnson SE. Growth factor modulation of equine trophoblast mitosis and prostaglandin gene expression. J Anim Sci. 2019 Feb 1;97(2):865-873. doi: 10.1093/jas/sky473. Carlisle LD, Memili E, Linford BL, Slater KA, Nicodemus MC. (2019) Relationship Between Gait Mechanics and the Speed of the Trot in the Weimaraner Dog Breed. Topics in Companion An Med. 35:26-30. Clark, K.L., Talton, O.O., Ganesan, S., Schulz, L.C., Keating, A.F. 2019. Developmental Origins of Ovarian Disorder: Impact of Maternal Lean Gestational Diabetes on the Offspring Ovarian Proteome in Mice. Biology of Reproduction. 101(4):771-781. Dickson, M.J., Kvidera, S.K., Horst, E.A., Wiley, C.E., Mayorga, E.J., Ydstie, J., Perry, G.A., Baumgard, L.H., Keating, A.F. 2019. Impact of chronic and increasing lipopolysaccharide exposure on production and reproductive parameters in lactating Holstein dairy cows. Journal of Dairy Science. 102(4):3569-3583. Dolecheck KA, Garcia-Guerra A, Moraes LE. 2019 Quantifying the effects of mastitis on the reproductive performance of dairy cows: A meta-analysis. Journal of Dairy Science, 102 (9): 8454–8477. Ealy AD and Seekford ZK. Symposium Review: Predicting pregnancy loss in dairy cattle. J Dairy Sci. 2019; 102:11798-11804. doi: 10.3168/jds.2019-17176. Ealy AD, Wooldridge LK and McCoski SR. Board Invited Review: Post-transfer consequences of in vitro-produced embryos in cattle. J Anim Sci. 2019 May 30;97(6):2555-2568. doi: 10.1093/jas/skz116. El Debaky HA, Kutchy NA, Ul-Husna A, Intriastuti R, Akhter S, Purwantara B, Memili E. (2019) Potential of water buffalo in world agriculture: challenges and opportunities. Applied Animal Science. 35:255-268. Evans H, Jousan D, Memili E, Beckman L, Nicodemus H. (2019) Life skill development and financial impact associated with a youth livestock sales program. Journal of Youth Development 14(3). Figueroa-Morales N, Dominguez-Rubio L, Ott TL, Aranson IS. 2019. Mechanical shear controls bacterial penetration in mucus. Sci Rep. 4;9(1):9713. doi: 10.1038/s41598-019-46085-z. PubMed PMID: 31273252; PubMed Central PMCID: PMC6609767. Fontes PL, Oosthuizen N, Ciriaco FM, Sanford CD, Canal LB, Pohler KG, Henry DD, Mercadante VR, Timlin CL, Ealy AD, Johnson SE, DiLorenzo N and Lamb GC. Impact of fetal vs. maternal contributions of Bos indicus and Bos taurus genetics on embryonic and fetal development. J Anim Sci. 2019 Apr 3;97(4):1645-1655. doi: 10.1093/jas/skz044. Garcia-Guerra A, Sala RV, Carrenho-Sala L, Baez GM, Motta JCL, Fosado M, Moreno JF, Wiltbank MC. 2019 Postovulatory treatment with GnRH on day 5 reduces pregnancy loss in recipients receiving an in vitro produced expanded blastocyst. Theriogenology, 141: 202-210. Gomes FP, Diedrich JK, Saviola AJ, Memili E, Moura AA, Yates JR. (2020) EThcD and 213 nm UVPD for Top-Down Analysis of Bovine Seminal Plasma Proteoforms on Electrophoretic and Chromatographic Time Frames. Anal Chem. doi: 10.1021/acs.analchem.9b03856. Gomez-Leon, V. E., O. J. Ginther, E. R. Araujo, J. D. Guimaraes, and M. C. Wiltbank. 2019. Hormonal mechanisms regulating follicular wave dynamics I: Comparison of follicle growth profiles under different physiological conditions in heifers. Theriogenology 123:194-201. Gomez-Leon, V. E., O. J. Ginther, J. D. Guimaraes, and M. C. Wiltbank. 2020. Hormonal mechanisms regulating follicular wave dynamics II: Progesterone decreases diameter at follicle selection regardless of whether circulating FSH or LH are decreased or elevated. Theriogenology 143:148-156. Habeeb et al. Effect of different dosages of PG-600 on ovulation and pregnancy rates in ewes during the breeding season. Transl. Anim. Sci. 2019.3:429–432. Habeeb et al. Ovulation, pregnancy, and lambing rates during nonbreeding season with or without exogenous gonadotropin stimulation. Clinical Theriogenology 2020.12:23-28.  Habeeb et al. Endometrial estrogen receptor, follicle stimulating hormone receptor, and mucin-1 expression during early diestrus in ewes following PG-600 administration. In review, Reproduction in Domestic Animals, April 30, 2020. Harl A, Westfall-Rudd D and Rhoads ML. 2019. Increased utilization of tactile activities in reproductive physiology laboratory sections to improve cognitive learning. NACTA J. 2019 63(2):341-343.  Harper MT, Oh J, Melgar A, Nedelkov K, Räisänen S, Chen X, Martins CMMR, Young M, Ott TL, Kniffen DM, Fabin RA, Hristov AN. 2019. Production effects of feeding extruded soybean meal to early-lactation dairy cows. J Dairy Sci. 102(10):8999-9016. doi: 10.3168/jds.2019-16551. Epub 2019 Aug 14. PubMed PMID: 31421886. Hines, E.A., Romoser, M.R., Kiefer, Z.E., Keating, A.F., Baumgard, L.H., Niemi, J., Haberl, B., Williams, N.H., Kerr, B.J., Touchette, K.J., Ross, J.W. 2019. The impact of dietary supplementation of arginine during gestation in a commercial swine herd: II. Offspring performance. Journal of Animal Science. 97(9):3626-3635.  Hines, E.A., Romoser, M.R., Kiefer, Z.E., Keating, A.F., Baumgard, L.H., Niemi, J., Gabler, N.K., Patience, J.F., Haberl, B., Williams, N.H., Kerr, B.J., Touchette, K.J., Ross, J.W. 2019. The impact of dietary supplementation of arginine during gestation in a commercial swine herd: I. Gilt reproductive performance. Journal of Animal Science. 97(9):3617-3625. Horst, E.A., Kvidera, S.K., Dickson, M.J., McCarthy, C.S., Mayorga, E.J., Al-Qaisi, M., Ramirez, A.A., Keating, A.F., Baumgard, L.H. 2019. Effects of continuous and increasing lipopolysaccharide infusion on basal and stimulated metabolism in lactating Holstein cows. Journal of Dairy Science. 102(4):3584-3597. Hughes CHK, Inskeep EK, Pate JL. 2020. Temporal changes in the corpus luteum during early pregnancy reveal regulation of pathways that enhance steroidogenesis and suppress luteolytic mechanisms. Biology of Reproduction. <a href="https://doi.org/10.1093/biolre/ioaa047">https://doi.org/10.1093/biolre/ioaa047</a>. Hughes CK, Pate JL. 2019. Luteolysis and the Corpus Luteum of Pregnancy. In: The Ovary 3rd Edition, PCK Leung and EY Adashi, eds., Academic Press, Elsevier, pp. 270-295.  ISBN 978-0-12-813209-8 Hughes CK, Maalouf SW Liu W-S, Pate JL. 2019. Molecular profiling demonstrates modulation of immune cell function and matrix remodeling during luteal rescue. Biology of Reproduction, 2019, 100(6), 1581–1596 doi:10.1093/biolre/ioz037 Hughes CHK, Bosviel R, Newman JW, Pate JL. 2019. Luteal lipids regulate progesterone production and may modulate immune cell function during the estrous cycle and pregnancy. Front. Endocrinol. 10:662.  doi: 10.3389/fendo.2019.00662 Jaskiewicz NJ, Townson DH (2019) Hyper-O-GlcNAcylation promotes epithelial-mesenchymal transition in endometrial cancer cells. Oncotarget 10(30):2899-2910 Khanal, P., Sanglard, L.P., Mayberry, K., Sommer, J., Poore, M.H., Poole, D.H., Serão N.V.L. 2019 Identification of differentially expressed genes for fescue toxicosis in high and low tolerant Angus cows. Epigenetics & Genomics DOI: 10.21203/rs.2.9998/v1 Kshitiz, Afzal J, Maziarz JD, Hamidzadeh A, Liang C, Erkenbrack EM, Kim HN, Haeger JD, Pfarrer C, Hoang T, Ott T, Spencer T, Pavličev M, Antczak DF, Levchenko A, Wagner GP. Evolution of placental invasion and cancer metastasis are causally linked. Nat Ecol Evol. 2019 Dec;3(12):1743-1753. doi: 10.1038/s41559-019-1046-4. Epub 2019 Nov 25. PubMed PMID: 31768023. Kutchy NA, Dogan S, Wang X, Topper E, Kaya A, Memili E. (2020) Application of Proteomics to Identify Fertility Markers in Angus Bull Sperm. In press; HAYATI Journal of Biosciences.  Kutchy NA, Menezes ESB, Ugur MR, Ulhusna A, ElDebaky H, Evans HC, Beaty E, Santos F, Tan W, Wills RW, Topper E, Kaya A, Moura AA, Memili E. (2019) Sperm Cellular and Nuclear Dynamics Associated with Bull Fertility. Anim Reprod Sci. Dec;211:106203. doi: 10.1016/j.anireprosci.2019.106203. Epub 2019 Oct 18. PMID: 31785643 Li H, Garner TB, Diaz FJ and Wong PK. A Multiwell Microfluidic Device for Analyzing and Screening Non-Hormonal Contraceptive Agents. Small. (2019) Jul;15(28):e1901910.  Macedo, G. G., R. D. Mingoti, E. O. S. Batista, B. M. Monteiro, L. M. Vieira, R. V. Barletta, M. C. Wiltbank, G. P. Nogueira, F. P. Renno, J. R. Maio, and P. S. Baruselli. 2019. Profile of LH release in response to intramuscular treatment with kisspeptin in Bos indicus and Bos taurus prepubertal heifers. Theriogenology 125:64-70. Mayorga, E.J., Kvidera, S.K., Horst, E.A., Al-Qaisi, M., Dickson, M.J., Seibert, J.T., Lei, S., Keating, A.F., Ross, J.W., Rhoads, R.P., Rambo, Z.J., Wilson, M.E., Baumgard, L.H. 2019 Effects of zinc amino acid complex on biomarkers of gut integrity and metabolism during and following heat stress or feed restriction in pigs.  Journal of Animal Science 96:4173-4185. Mayorga, E.J., Kvidera, S.K., Seibert, J.T., Horst, E.A., Abuajamieh, M., Al-Qaisi, M., Lei, S., Ross, J.W., Johnson, C.D., Kremer, B., Ochoa, L., Rhoads, R.P., Baumgard, L.H.  2019. Effects of dietary chromium propionate on growth performance, metabolism, and immune biomarkers in heat-stressed finishing pigs. J Anim Sci. 97:1185-1197. Mayorga E.J., Renaudeau, D., Ramirez, B.C., Ross, J.W., Baumgard, L.H. 2019.  Heat stress adaptation in pigs. Animal Frontiers 9:54-61. Memili E, Moura A, Kaya A. (2020) Metabolomes of Sperm and Seminal Plasma Associated with Bull Fertility. Animal Reproduction Science. doi: 10.1016/j.anireprosci.2020.106355. Menezes E, Velho A, Santos F, Dinh T, Kaya A, Topper E, Moura A, Memili E. (2019) Uncovering Sperm metabolome to discover biomarkers for bull fertility. BMC Genomics. 20(1):714. doi:10.1186/s12864-019-6074-6. Menezes ESB, Badial PR, El-Debaky H, Ulhusna A, Ugur MR, Kaya A, Topper E, Bulla C, Grant KE, Bolden-Tiller O, Moura AA, Memili E. (2019) Sperm miR-15a, miR-29b, miR-34a are associated with male fertility. doi: 10.1111/and.13412. Mezera, M. A., C. S. Hamm, C. A. Gamarra, R. S. Gennari, A. B. Prata, R. Sartori, and M. C. Wiltbank. 2019. Profiles of prostaglandin F2alpha metabolite in dairy cattle during luteal regression and pregnancy: implications for corpus luteum maintenancedagger. Biol Reprod 101:76-90. Monn, R. E., Mackey, J.C., Mayberry, K. J., Poole, R. K., Dudley, H., Alley, M., Poole, D. H. 2019. A two injection of Prostaglandin F2α pre-synchronization treatment decreases conception rates of developing replacement heifers. Translational Animal Science Volume 3, Issue 1, 1 January 2019, Pages 456–463, https://doi.org/10.1093/tas/txy136 Navarrete FA, Aguila L, Martin-Hidalgo D, Tourzani DA, Luque GM, Ardestani G, Garcia-Vazquez FA, Levin LR, Buck J, Darszon A, Buffone MG, Mager J, Fissore RA, Salicioni AM, Gervasi MG, Visconti PE. Transient Sperm Starvation Improves the Outcome of Assisted Reproductive Technologies. Front Cell Dev Biol. 2019 Nov 5;7:262. doi: 10.3389/fcell.2019.00262. Negron-Perez VM, Fausnacht DW and Rhoads ML. Invited Review: Management strategies capable of improving the reproductive performance of heat-stressed dairy cattle. J Dairy Sci. 2019 Dec;102(12):10695-10710. doi: 10.3168/jds.2019-16718. Niles, A. M., H. P. Fricke, P. D. Carvalho, M. C. Wiltbank, L. L. Hernandez, and P. M. Fricke. 2019. Effect of treatment with human chorionic gonadotropin 7 days after artificial insemination or at the time of embryo transfer on reproductive outcomes in nulliparous Holstein heifers. J Dairy Sci 102:2593-2606. Ozbek M, Hitit M, Ergun E, Ergun L, Beyaz F, Erhan F, Yildirim N, Kandil B, Ozgenc O, Memili E. (2020). Expression Profile of Toll-Like Receptor 4 in Rat Testis and Epididymis throughout Postnatal Development. Andrologia. doi:10.1111/and.13518. Pereira, M. H. C., M. C. Wiltbank, T. G. Guida, F. R. Lopes, Jr., B. I. Cappellozza, and J. L. M. Vasconcelos. 2020. Evaluation of presynchronization and addition of GnRH at the beginning of an estradiol/progesterone protocol on circulating progesterone and fertility of lactating dairy cows. Theriogenology 147:124-134. Pillai VV, Kei TG, Reddy SE, Das M, Abratte C, Cheong SH and Selvaraj V. (2019). Induced pluripotent stem cell generation from bovine somatic cells indicates unmet needs for pluripotency sustenance. Animal Science Journal. 90(9):1149-1160. Plewes MR, Hou X, Zhang P, Wood JR, Cupp AS, Lu X, Wang C, and Davis JS (2019) Yes-associated protein (YAP) is required for proliferation and function of bovine granulosa cells. Biol Reprod 101: 1001-1017, PMID: PMC6877782  Poole D.H. 2019 Impacts of fescue toxicosis on the success of reproductive technologies. Proceedings in Applied Reproductive Strategies in Beef August 20-21, 2019 Poole RK, Womble CM, Poore MH, Poole DH, Pickworth CL. 2019 Effects of endophyte-infected tall fescue seed and protein supplementation on stocker steers: I. Growth performance and hemodynamic responses. J Anim Sci. 2019 Sep 3;97(9):3776-3785. doi: 10.1093/jas/skz249. PMID: 31359059 Poole RK, Brown AR, Poore MH, Pickworth CL, Poole DH. 2019 Effects of endophyte-infected tall fescue seed and protein supplementation on stocker steers: II. Adaptive and innate immune function. J Anim Sci. 2019 Oct 3;97(10):4160-4170. doi: 10.1093/jas/skz250. PMID: 31353402 Poole RK, Poole DH. 2019. Impact of Ergot Alkaloids on Female Reproduction in Domestic Livestock Species. Toxins (Basel). Jun 21;11(6):364. doi: 10.3390/toxins11060364. PMID: 31234268 Free PMC article. Invited Review. Poole RK, Devine TL, Mayberry KJ, Eisemann JH, Poore MH, Long NM, Poole DH. 2019. Impact of slick hair trait on physiological and reproductive performance in beef heifers consuming ergot alkaloids from endophyte-infected tall fescue. J Anim Sci. Apr 3;97(4):1456-1467. doi: 10.1093/jas/skz024. Ramirez, B.C., Xin, H., Halbur, P.G., Beermann, D.H., Hansen, S.L., Linhares, D.C.L., Peschel, J.M., Rademacher, C.J., Reecy, J.M., Ross, J.W., Shepherd, T.A., Koltes, J.E. 2019. At the intersection of industry, academia, and government: How do we facilitate productive precision livestock farming in practice? Animals (Basel) 9(9). Rhoads ML. Effects of periconceptional heat stress on primiparous and multiparous daughters of Holstein dairy cows. Theriogenology. 2020 Mar 11. pii: S0093-691X(20)30181-3. doi: 10.1016/j.theriogenology.2020.03.015. Rister AL, Bidne KL, Wood JR, and Dodds ED (2019) Microscale serum extraction method for the simultaneous analysis of corticosterone and lipids. Anal Methods, 11: 5746-5749, PMID: 32148562 Schalich KM, Herren A and Selvaraj V. (2020). Analysis of differential strategies to enhance detection of low-abundance protein in the bovine serum proteome. Animal Science Journal. Accepted, In press. Schultz, R., Serao, N.L., Ross, J.W. 2019.  Genetic improvement of livestock, from conventional breeding to biotechnological approaches. In: Animal Agriculture: Sustainability, Challenges and Innovations. Eds. Bazer, F.W., Wu, G.  In Press (Elsevier).  Seibert, J.T., Adur, M.K., Schultz, R.B., Thomas, P.Q., Kiefer, Z.E., Keating, A.F., Baumgard, L.H., Ross, J.W. 2019. Differentiating between the effects of heat stress and lipopolysaccharide on the porcine ovarian heat shock protein response. Journal of Animal Science. 97(12):4965-4973.  Silveira, P. A. S., W. R. Butler, T. C. da Silva, C. C. Barros, M. N. Correa, and A. Schneider. Association of polymorphisms in the IGF-I, GHR, and STAT5A genes with serum IGF-I concentration and reproductive performance of Holstein dairy cows. Anim Reprod.Sci. 211(https://doi.org/10.1016/j.anireprosci.2019.106206):106206, 2019. Snider AP and Wood JR (2019) Obesity induces ovarian inflammation and reduces oocyte quality. Reproduction 158: R79-R90, PMID: 30999278 Sun T and Diaz FJ. Ovulatory signals alter granulosa cell behavior through YAP1 signaling. Reproductive Biology and Endocrinology: (2019) 17 (1), 113. Timme KR and Wood JR (2019) Use of single molecule fluorescent in situ hybridization (SM-FISH) to quantify and localize mRNAs in murine oocytes. J. Vis. Exp. 146, PMID: 31081819 Tenley SC, Spuri-Gomes R, Rosasco SL, Northrop EJ, Rich J, McNeel AK, Summers AF, Miles JR, Chase CC, Lents CA, Perry GA, Wood JR, Cupp AS, Cushman RA (2019) Maternal age influences the number of primordial follicles in the ovaries of yearling angus heifers, Animal Reproduction Science 200:105-112 Ugur MR, Dinh T, Hitit M, Kaya A, Topper E, Didion B, Memili E. (2020) Amino Acids of Seminal Plasma Associated with Freezability of Bull Sperm. Frontiers in Cell and Developmental Biology 7(347): 1-14. Ugur MR, Saber Abdelrahman A, Evans HC, Gilmore AA, Hitit M, Arifiantini RI, Purwantara B, Kaya A, Memili E. (2019) Advances in cryopreservation of bull sperm. Front Vet Sci. doi: 10.3389/fvets.2019.00268. Ugur MR, Kutchy NA, Menezes EBS, Ulhusna A, Haynes BP, Uzun A, Kaya A, Topper E, Moura AA, Memili E. (2019) Retained Acetylated Histone Four in Bull Sperm Associated with Fertility. Frontiers in Veterinary Science 6(23): 1-10. Vailes MT, McCoski SR, Wooldridge LK, Reese ST, Pohler KG, Roper DA, Mercadante VR and Ealy AD. Post-transfer outcomes in cultured bovine embryos supplemented with epidermal growth factor, fibroblast growth factor 2, and insulin-like growth factor 1. Theriogenology. 2019 Jan 15;124:1-8. doi: 10.1016/j.theriogenology.2018.09.023. Velho A, Wang H, Koenig L, Grant KE, Menezes E, Kaya A, Moura A, Memili E. (2019) Expression dynamics of Integrin Subunit Beta 5 in bovine gametes and embryos imply functions in male fertility and early embryonic development. Andrologia. 00:e13305. DOI: 10.1111/and.13305. Wang L; Hand JM, Fu L, Smith GW and Yao J. 2019. DNA methylation and miRNA-1296 act in concert to mediate spatiotemporal expression of KPNA7 during bovine oocyte and early embryonic development. BMC Developmental Biology. 19 (1), 23. DOI: 10.1186/s12861-019-0204-x. Wang J, Koganti PP and Yao J. 2020. Systematic identification of long intergenic non-coding RNAs expressed in bovine oocytes. Reproductive Biology and Endocrinology. 18(1) 13. DOI: 10.1186/s12958-020-00573-4. Wooldridge LK and Ealy AD. Interleukin-6 increases inner cell mass numbers in bovine embryos. BMC Dev Biol. 2019 Feb 1;19(1):2. doi: 10.1186/s12861-019-0182-z. Wooldridge LK and Ealy AD. Zinc Supplementation during in vitro bovine embryo culture increases inner cell mass and total cell numbers in bovine blastocysts. J. Anim. Sci. 2019; 97(12):4946-4950. doi: 10.1093/jas/skz351. Wooldridge LK, Johnson SE and Ealy AD. Interleukin-6 requires JAK to stimulate inner cell mass expansion in bovine embryos. Reproduction. 2019 Oct;158(4):303-312. doi: 10.1530/REP-19-0286. Zhang Z, Zhao LD, Johnson SE, Rhoads ML, Jiang H and Rhoads RP. Oxytocin is involved in steroid hormone-stimulated bovine satellite cell proliferation and differentiation in vitro. Domest Anim Endocrinol. 2019 Jan;66:1-13. doi: 10.1016/j.domaniend.2018.07.003.   Theses/Dissertations Battista SEB, Garcia-Guerra A. Associations between morphometric characteristics of the reproductive tract and fertility in beef cattle. OSU MS thesis. Heather L. Baldwin. Investigation of bovine conceptus galectins and the effect of bovine conceptus secretory proteins on the endometrial epithelial transcriptome. (MS Thesis) Francesca Anastasia Gambonini, M.S. Effects of an Extruded Flaxseed Supplement on Transition Cow Milk Production and Reproductive Function and Effects of Early Pregnancy on Mediators of Immune Tolerance in Dairy Heifers. Master of Science Thesis. The Pennsylvania State University. Lindsay F. Grose. The effect of bovine galectin-1, a conceptus secretory protein, on the endometrial transcriptome. (MS Thesis) James Hester, PhD.  The impact of zinc deficiency during oogenesis, follicle assembly and growth. Camilla Hughes, Phd. Molecular Changes in the Corpus Luteum During Maternal Recognition of Pregnancy. PhD Dissertation. The Pennsylvania State University Shaina Jachter, B.S. 06/2020. Undergraduate Honor’s Thesis: “Effects of Kit-Ligand on Survival and Growth of Bovine Multilayer Follicles in Culture.” Honors Thesis Advisor C. Bishop. Lundberg, Townson DH (2019) Investigation of Transforming Growth Factor-α and its potential role in promoting ovarian follicular dominance. UVM MS Thesis Zackary K. Seekford (2020). Interleukin-6 Supplementation Improves Post-Transfer Embryonic and Early Fetal Development of In Vitro Produced Bovine Embryos (MS Thesis). Wilson Simmons, M.S. 06/2020. Master’s Thesis: “Epidermal Growth Factor and Leukemia Inhibitory Factor Supplementation in Matrix-free, Three-Dimensional Bovine Follicle Culture.” Major Professor C. Bishop. Lydia K. Wooldridge (2020). Supplementing Bovine Embryo Culture Media to Improve the Production and Quality of In Vitro Produced Bovine Embryos (PhD Dissertation).    Active collaboration within the group OSU and WI:  evaluated the effect of dose of GnRH at initiation of a 5-day synchronization protocol for FTET. NY, PA, NH, VT and WV:  Samples collected to investigate associations between fertility outcomes and SNP in candidate genes from dairy cows. PA and WV: Samples shared for analysis of molecular regulators of luteal function.    In addition to Hatch Multistate Funds, these studies were supported by: Lee Rumberger and Family Endowment to JLP CFAES Honors Will C. Hauk Endowment Grant – Undergraduate student - The Ohio State University Iowa Pork Producers Association            Iowa State University Bailey Career Development Award John L. Pratt Animal Nutrition Program NIH   National Institute of Child Health and Human Development National Institute of Environmental Health Science National Institute of Food and Agriculture National Pork Board North Carolina Cattlemens Association Assessment North Carolina Agricultural Foundation Inc. OARDC SEEDS Early career investigator – The Ohio State University OARDC SEEDS grant Competition – Graduate Student grant – The Ohio State University OSU Agricultural Research Foundation RTF funds from Dean to PI RTF funds from PIs Theriogenology Foundation Research Grant USDA-NIFA-AFRI Virginia Agricultural Council Research Funding

Impact Statements

Low pregnancy rates in beef and dairy cows result in increased costs to producers associated with increased rebreeding or culling of animals, and reduced likelihood of recouping heifer development costs. Reduced fertility is also a major cause of reduced milk (dairy) and meat (beef) production which impacts the food supply. The focus of the NE1727 multistate project “Influence of ovary, uterus, and embryo on pregnancy success in ruminants”, is to understand how altered ovarian function, impaired oocyte quality, and disruptions of conceptus-uterine interactions contribute to infertility, and devise management strategies that will overcome these factors in order to optimize the chances that animals successfully achieve a pregnancy. To achieve this goal, the following three objectives were performed: (1) Determine the impact of altered ovarian function on reproductive performance; (2) Identify alterations in embryo development and uterine and CL function associated with declining pregnancy establishment; and (3) Identify changes in genetics and reproductive management that lead to improved pregnancy rates. As investigators in NE1727, we have used our collective expertise to broaden the current understanding of the mechanisms by which internal and external forces modify these important physiological processes. Furthermore, this information about core biological processes was used to rationally develop new and innovative tools to increase cyclicity and breeding efficiency, and reduce early embryonic failures. The outcome of these accomplishments was improving reproductive efficiency of both dairy and beef cattle operations in the United States.

Date of Annual Report: 11/02/2021

Report Information

Annual Meeting Dates: 08/12/2021 - 08/13/2021
Period the Report Covers: 10/01/2020 - 09/30/2021

Participants

Michelle Rhoads, Phillip Bridges, David Townson, Joanne Fortune, Milo Wiltbank, Aileen Keating, Alan Ealy, Cecily Bishop, Daniel Mathew, Erdogan Memili, Jennifer Wood, Jianbo Yao, Joy Pate, Julio Giordano, Paul Tsang, Rafael Fissore, Bob Dailey
Rodolfo Cardoso, Ron Butler, Troy Ott, Dan Poole, Adam Beard, Soon Hon Cheong, Alvaro Garcia-Guerra, Athena Ren, Vimal Selvaraj.

Brief Summary of Minutes

Minutes Attachment:

2-NE1727 Minutes. 2021 Virtual Meeting_JG08122021.docx

Accomplishments

The following has been accomplished: Investigated the effect of supplementing cattle with a vitamin-mineral mix containing sodium selenite, an organic form, or an isomolar 1:1 mix of ISe and OSe (MIX), and demonstrated that the MIX form (versus OSe or ISe alone) increases early luteal phase systemic progesterone (P4), which advances embryonic development. Investigated the mechanism responsible for this MIX-induced increase in P4.  Overall, it appears that the MIX-induced increase in early luteal phase P4 is not directly mediated by an increase in the expression of key steroidogenic transcripts, but by an increase in cholesterol uptake, through at least the low-density lipoprotein receptor (LDLR). Widespread regions of the southeast United States have soils deficient in selenium (Se), necessitating Se supplementation to grazing cattle. Thus, investigated the effect of form of supplemental Se on the transcriptome of the bovine corpus luteum (CL) with a goal of elucidating form of Se-regulated luteal processes affecting fertility. Our results suggested that the MIX-induced increase in early luteal phase P4 is due to an increase in cholesterol availability and that the form of dietary Se affects immune function of the CL. Completed a study on impact of brief vitamin A and D supplementation during synchronization protocols in lactating dairy cows. While we did successfully elevate serum vitamin D, no differences were observed in pregnancy rates following timed artificial insemination. A parallel in vitro study of 25(OH) vitamin D in presence and absence of low level trans-retinoic acid (vitamin A, both active forms) supplementation in vitro found vitamin D alone at low concentrations improved embryo characteristics in culture. However, vitamin D and A combined improved fertilization rates in vitro. Compared follicle selection during endogenous gonadotropin pulses versus during ablation of pulses with Acyline (GnRH antagonist) and luteinizing hormone (LH) action replaced with nonpulsatile human chorionic gonadotropin (hCG) treatment in Trio carriers (n = 28) versus noncarriers (n = 32). Acyline +hCG decreased (P< 0.0001) the number of DFs from 2.7 ± 0.2 to 1.3 ± 0.2 in Trio carriers, with most heifers having only one DF. This demonstrates the necessity for LH in acquisition of dominance in Trio carriers (&sim;6.5 mm) and noncarriers (&sim;8.5 mm) and provides evidence for a role of GnRH-induced FSH/LH pulses in selection of multiple DFs in Trio carriers and possibly other physiologic situations with increased ovulation rate. Tested the hypotheses that luteolysis after induced pregnancy loss (IPL) will be temporally associated with a decrease in ipsilateral uterine artery vascular perfusion and an increase in PGFM pulse frequency and amplitude. Pregnant non-lactating beef cows were randomly assigned to undergo IPL at day 35 of gestation (IPL, n=10) or remain untreated (CON, n=6). Corpus luteum (CL) volume and embryo/fetal viability were evaluated by transrectal B-mode ultrasonography daily from gestation day 34 until luteolysis has been identified. Luteolysis was defined as the day before serum progesterone decreased to ≥50% less than the average for the four maximum progesterone concentrations. Uterine artery vascular perfusion will be evaluated using spectral-doppler ultrasonography. Blood samples were collected at bihourly intervals and quantification of plasma progesterone and PGFM is currently being performed. Determined the effect of (i) Pre-synchronization (Presynch) prior to a 6-d CO-Synch protocol and (ii) circulating P4 on ovulatory response, estrous expression and pregnancy per AI (P/AI) in beef heifers. Presynch heifers had larger follicle diameter at GnRH treatment, greater ovulatory response, greater estrus expression and expressed estrus earlier compared to controls. There was a treatment by CIDR interaction on estrous expression, whereas a lesser percentage of control heifers with new CIDR showed estrus compared to all other groups. Heifers treated with a used P4 device tended to have greater P/AI (52.1%; 61/117) than those with a new CIDR (40.5%; 47/116). Thus, Pre-synchronization prior to initiation of a 6-d CO-Synch increased follicle diameter, ovulatory response, and estrous expression, however, did not affect fertility. The earlier onset of estrus in Presynch heifers warrants further study on timing of AI. Evaluated the effect of increasing progesterone (P4) device insertion from 5 to 6 days during a TAI protocol on ovarian dynamics and fertility in suckled beef cows (n=867) at six locations. Cows in the 6-day group tended (P=0.09) to have a larger maximum follicle diameter (16.2±0.4mm) than cows in the 5-day group (15.0±0.6mm). In addition, CL volume on d7 was greater (P=0.01) in the 6-day (4302±495mm3) than 5-day (2406±455mm3) group. There were, however, no differences (P=0.15) in the interval between CIDR removal and ovulation. Cows assigned to the 6-day group had greater (P < 0.05) estrus expression [56.1% (215/385) vs 41.5% (161/388)] and pregnancy rate [54.6% (236/438) vs 47.4% (207/435)] than cows in the 5-day group. In conclusion, extension of the period of P4 device insertion in a prolonged proestrus protocol increases preovulatory follicle size, estrous expression, and fertility to TAI.   Evaluated the effect of FSH dose and circulating P4 on ovarian superstimulation responses, before ovum pick-up (OPU) on IVP in pregnant heifers. Heifers treated with 300 mg of p-FSH had a greater (P < 0.05) number of medium size follicles, total number of follicles, total COCs recovered, number of viable COCs, and number of blastocysts produced than heifers treated with 160 mg of p-FSH. In addition, heifers treated with 300 mg of p-FSH had greater cleavage rate (P = 0.03). Circulating P4 did not affect (P > 0.10) any of the outcomes evaluated nor was there an FSH dose by circulating P4 interaction (P > 0.05). In conclusion, increasing the dose of p-FSH prior to OPU from 160 to 300 mg results in improved embryo production in pregnant donors, regardless of progesterone concentrations at the time of OPU.   Determined the effect of length of the superstimulatory treatment period prior to OPU on ovarian response and IVP in Holstein heifers. Lengthening the FSH treatment period resulted in greater (P = 0.01) number of large follicles, however, the number of small, medium and total follicles was not different between groups. Number of total recovered oocytes, viable oocytes, cleaved oocytes, as well as recovery rate, percent viable oocytes and cleavage rate were not affected. Similarly, number of blastocysts produced per heifer and blastocyst rate were not different (P > 0.9) among groups. In conclusion, lengthening the period of FSH treatment by 1 d increases the number of large follicles at OPU, however, does not improve overall ovarian response, oocyte recovery nor embryo production.  Evaluated the effect of length of the FSH superstimulatory treatment on ovarian response and embryo production in sheep. Embryo yield were not affected by FSH administration in either 6 or 8 doses during the first follicular wave in sheep. Compared the performance of beef cattle submitted to resynchronization for 2nd service at 16 or 25 d after 1st service. Resynchronization at d25 in beef cattle resulted in improved reproductive performance compared to resynchronization at d16, however, it involves a greater interval between 1st and 2nd service, 34 vs 25 d, respectively. Gilts have been treated to determine an additive impact of thermal stress on the ovarian response to chemical exposure. Phenotypic and endocrine data has been generated. LC-MS/MS to identify alterations in the ovarian proteome have been completed, western blotting to investigate specific target proteins have been completed. Two manuscripts are in preparation. Investigated the impact of thermal stress on the ovarian prolactin signaling pathway in heat stressed gilts. Completed proteomic analysis of ovaries from mice exposed to perfluorooctonic acid (PFOA) or dimethylbenz[a]anthracene or Glyphosate (GLY). Collected phenotypic data from each of these separate experiments. In addition, both lean and obese mice were included in each study and the impact of altered metabolic status has been determined. Similar to OVCAR8 cells, CCN1 expression in two other cancer cell lines (SK-OV-3 and OVCAR3) that have constitutively active STAT3 was induced by serum. Further, CCN1 expression was increased by treatment with calcium ionophore and STAT3 siRNA. In OVCAR8 cells, the constitutive activation of STAT3 negatively affects CCN1 expression. However, when STAT3 is knocked down with a STAT3 siRNA, CCN1 expression is increased. Thus, could the constitutive (high) STAT3 environment play a role in the ability of PGF2α to regulate CCN1? When OVCAR 8 cells were treated with STAT3 siRNA, two concentrations of PGF2α (0.1 and 0.5μM) decreased CCN1 expression. More experiments are needed to determine the cellular mechanisms by which this decrease in CCN1 occurs. In steroidogenic bovine luteal cells, our previous findings suggest little to no involvement of the protein kinase A (PKA) pathway in the regulation of CCN1 expression. Evidence from studies in endothelial cells and a variety of cancer cells implicates RhoA as a regulator of CCN1 expression. Here, we showed that treatment with the Rho Activator II increased CCN1 expression in steroidogenic bovine luteal cells. Further, treatment with thrombin, an activator of RhoA, also increased CCN1 expression in these cells. Evaluated if maternal nutrition during the second and third trimester of gestation and dietary energy intake during the juvenile period interact to affect concentrations of neuropeptide Y (NPY) in the third ventricle cerebrospinal fluid (CSF) after fasting (basal) and in response to peripheral leptin administration in heifer offspring. Treatment with leptin had no effect (P > 0.10) on NPY concentration in any group. Therefore, neither the interaction of pre- and postnatal nutrition nor exogenous leptin affected third ventricle NPY tone in an experimental model. Identified 29 follicular fluid metabolites that had positive relationship with pre-ovulatory follicle diameter at the time of GnRH injection to induce ovulation (better understand metabolic impacts of induced ovulation in beef cows). Identified 1,387 and 2,765 mRNA transcripts with unique expression and 2,553 and 1,745 mRNA transcripts with upregulated expression in in vivo matured bovine oocytes or cumulus cells, respectively (improve understanding of oocyte and cumulus function during in vivo maturation). Identified progesterone stimulated bovine endometrial epithelial and stroma fibroblast cell specific mRNAs (better understand uterine support of early embryonic development). Tested for an advantage of progesterone stimulated bovine endometrial epithelial and stroma fibroblast cell conditioned media on in vitro produced (IVP) bovine embryo development. Identified 21 serum metabolites from blood collected in the pre-ovulatory time period, that differed among beef cows with thin, moderate, and obese body condition scores. This will help better understand the impact of body condition on fertility. Demonstrated that O-GlcNAcylation is readily evident in bovine granulosa cells (GCs) of both small and large follicles. Also, disruption of O-GlcNAcylation in granulosa cells impairs cell proliferation. Demonstrated that granulosa cells from small follicles have greater glycolytic capacity than GCs from large follicles; but disruption of the hexosamine biosynthesis pathway and O-GlcNAcylation has little to no effect on GC metabolic activity. Demonstrated that transvaginal oocyte aspiration of medium-sized follicles every third day for 30 days does not disrupt ovarian follicular development of early lactation dairy cattle.  Likewise, collection of uterine biopsies alongside every other transvaginal oocyte aspiration does not affect cyclicity. The role of interleukin-6 as an embryokine has been further established by observing that treating bovine embryos with this factor will alter the early development of the fetus so these pregnancies more closely resemble pregnancies from cows that were artificially inseminated.               Generated evidence that the primary actions of IL-6 are to 1) enable development of embryos cultured individually, 2) improve development of the inner cell mass (ICM) through a JAK-dependent mechanism, and 3) promote primitive endoderm development through a JAK-dependent mechanism. Identified profiles of gut microbial families that are associated with increased ovarian inflammation and reduced reproductive performance in beef cows. Determined the mechanistic effects of inflammation and oxidative stress on bovine somatic cell steroidogenesis, oocyte maturation and pre-implantation embryo development. Maternal factors are essential aspects of oocyte competence, which orchestrate early embryonic development. ZNFO is a Krüppel-associated box domain (KRAB) containing zinc finger transcription factor, which is exclusively expressed in bovine oocyte. Previous studies have demonstrated that ZNFO is essential for early embryonic development. We identified ZNFO as the core promoter of an oocyte-specific maternal effect gene. Long noncoding RNAs (lncRNAs) regulate various cellular processes including modulation of gene expression and epigenetic status. We identified differential expression of oocyte-specific lncRNAs in oocytes derived from small and large follicles. Factors present in the oocyte and surrounding follicular cells aid in the attainment of oocyte competence. Agouti-signaling protein (ASIP) is a known regulator of melanocyte function through binding to melanocortin receptors including MC1R and MC4R. We characterized expression of ASIP gene in ovarian tissues, and during oocyte maturation and early embryogenesis. We are understanding the mechanisms involved in selection of a single dominant follicle in cattle. This will be key for the rational design of new reproductive management strategies and reproductive biotechnologies. We have discovered the mechanism that produces increased ovulation rate in carriers of the Trio allele.  This mechanism may also underlie double ovulation in dairy cattle and in other species.  This should allow us to manipulate the ovulation rate in cattle and perhaps other species. We have now determined how to practically utilize Trio to reliably produce bilateral twins in beef cattle. This may be of great practical value to beef cattle producers in the future. We now have clearly demonstrated that the corpus luteum is maintained during early pregnancy by suppression of pulses of prostaglandin F2a by interferon-tau, based on our measurements of PGFM surges. We have also discovered that there is a second period of corpus luteum maintenance after Day 25 when interferon-tau secretion ends and PGF pulses are initiated. The resolution of the mechanisms that allow maintenance of the corpus luteum during this second period will be key for reducing pregnancy loss in cattle. We found that increasing circulating progesterone can reduce pregnancy loss in recipients of in-vitro produced embryos. This provides an easy method to reduce pregnancy loss by treatment with either GnRH or hCG. We demonstrated that an ear-attached automated estrus detection system (Smartbow; Zoetis Inc.) monitored physical activity and rumination time was effective at detecting cows in estrus and generated few false positive alerts when accounting for ovulation. Documented differences in behavioral, physiological, and performance parameters monitored by automated wearable and non-wearable sensor before insemination in lactating dairy cows that become pregnant or not to first service, but differences are parity-specific. Developed and evaluated models to predict the likelihood of pregnancy at first service in multiparous cows. These models could be approximated by using early lactation health records, body condition, hormone concentrations, and calving season data. We created and tested an intravaginal hormone releasing device prototype that can be programmed to automatically release PGF for successful induction of luteal regression in lactating dairy cows. We demonstrated that age at first calving affected the proportion of primiparous cows inseminated at detected estrus for first AI and first service P/AI, but did not affect the pregnancy and herd exit dynamics up to 200 DIM for cows managed with a program that favored insemination at detected estrus rather than TAI. We observed that age at first calving affected the reproductive performance of primiparous lactating Holstein cows managed with a program that favored TAI because cows with younger age at first calving had improved reproductive performance.  Demonstrated that the profitability of replacement dairy heifers during a 15-mo period after the beginning of the insemination period increases as age at first calving decreases. Observed that reproductive programs that used AI at detected estrus after automated estrus alerts for first service led to similar reproductive performance than an all-TAI program and using automated estrus alert data helps identify cows more likely to be inseminated at detected estrus and become pregnant to first service. Demonstrated that primiparous cows managed with a reproductive program that prioritized TAI or a program that prioritized AI at detected estrus had similar cash flow for up to 18-mo after calving in their first lactation. We studied pluripotency in cattle and simultaneously, simple ways to determine fertilization success and pregnancy in a breeding program. In this project, we explored ways to develop induced pluripotent stem cells from cattle and identify biomarkers in bovine serum that can be used in the diagnosis of pregnancy and pregnancy failures.  We identified an extensive profile of bioactive proteins secreted by the uterine endometrium at periods relevant to early embryo development. This contributed to our efforts to understand the uterine microenvironment and factors controlling successful early development leading to initiation of implantation.  Through careful examination of cells in circulation in cows pregnant with male and female calves we established that fetal microchimerism does not occur in cattle. This showed that fetal sex cannot be identified using a simple blood draw (as can be done in humans and rodents). Therefore, identification of fetal sex for use in management decisions through analysis of blood is not possible in cattle. Identified changes in YAP signaling during oocyte maturation. Identified potentially novel pathways important for maturation of the cumulus-oocyte complex. MicroRNA-1246 is abundant in luteal cell-derived extracellular vesicles, and can directly impact function of T Cells. Let-7 microRNA are highly abundant in the bovine corpus luteum. Let 7 microRNA can alter mitochondrial function in, and progesterone production by, luteal cells. Collaboration of dataset analyses between Penn State and University of Wisconsin allowed for the identification of common pathways that are likely regulated in the CL or pregnancy. Supplemental omega-3 fatty acids during the transition period improved reproductive performance in dairy heifers, but not cows and differentially affected immune and metabolic markers in dairy cows and heifers. Pregnancy in dairy cattle is associated with activation of molecules associated with immune tolerance in peripheral blood leukocytes.

Publications

Peer-reviewed journal articles reporting research from this project Andrade JPN, Gomez-Leon VE, Andrade FS, Carvalho BP, Lacouth KL, Garcia FZ, Jacob JCF,Sales JNS, Wiltbank MC, Mello MRB. Development of a novel 21-day reinsemination program, ReBreed21, in Bos indicus heifers. Theriogenology 2020; 155:125-131.  10.1016/j.theriogenology.2020.04.021 Arbeithuber B, Hester J, Cremona MA, Stoler N, Higgins B, Anthony K, Chiaromonte F, Diaz FJ, Makova KD. Highly accurate mitochondrial DNA sequencing reveals age-dependent accumulation of mutations in germline and somatic mouse tissues. PLOS Biology (2020):18 (e3000745). Bidne KL, Rister AL, McCain AR, Hitt BD, Dodds ED, and Wood JR. (2020) Maternal obesity alters placental lysophosphatidylcholine, lipid storage, and the expression of genes associated with lipid metabolism. Biol Reprod 104: 197-210, PMID: 33048132 Carr, S.N., Y. Jia, B. Crites, C. Hamilton, J.L. Edwards, J.C. Matthews and P.J. Bridges.  2020.  Form of supplemental selenium in vitamin-mineral premixes differentially affects early luteal and gestational concentrations of progesterone, and postpartum concentrations of prolactin in beef cows.  Animals.  10:1-11. Chaney H. L., L. F. Grose, G. Charpigny, S. K. Behura, I. M. Sheldon, J. G. Cronin, T. E. Spencer, P. Lonergan, D. J. Mathew. Conceptus-induced, interferon tau dependent gene expression in bovine endometrial epithelial and stromal cells. Biol Reprod. 2020. 104:669-683. doi:10.1093/biolre/ioaa226. Clark, K.L., Keating, A.F. 2020. Ataxia Telangiectasia Mutated coordinates the ovarian DNA repair and atresia-initiating response to phosphoramide mustard. Biology of Reproduction. 102(1):248-260. Clark, K.L., Roach, C.M., Keating, A.F. 2020. Obesity alters ovarian DNA damage response and apoptotic proteins. Reproduction. 160(5): 751-760.  Corredor, F.A., Pereira Sanglard, L., Leach, R.J., Ross, J.W., Keating, A.F., Serão, N.V.L. 2020. Genetic and genomic characterization of vulva size traits in Yorkshire and Landrace gilts. BMC Genetics. 21: 28.  Domingues RR, Ginther OJ, Gomez-Leon VE, Wiltbank MC. Up-regulation of endometrial oxytocin receptor is associated with the timing of luteolysis in heifers with two and three follicular waves. Biol Reprod 2020; 102:316-326.  10.1093/biolre/ioz165 Domingues RR, Ginther OJ, Toledo MZ, Wiltbank MC. Increased dietary energy alters follicle dynamics and wave patterns in heifers. Reproduction 2020; 160:943-953.  10.1530/Rep-20-0362 Drum JN, Wiltbank MC, Monteiro PLJ, Prata AB, Gennari RS, Gamarra CA, Canavessi AMO, Sartori R. Oxytocin-induced prostaglandin F2-alpha release is low in early bovine pregnancy but increases during the second month of pregnancy. Biol Reprod 2020; 102:412-423.  10.1093/biolre/ioz169 Ealy AD, Speckhart SL, Wooldridge LK. Cytokines that serve as embryokines in cattle. Animals 2021, 11(8), 2313; https://doi.org/10.3390/ani11082313 Fontes PLP, Oosthuizen N, Ciriaco FM, Sanford CD, Canal LB, Cooke RF, Pohler KG, Henry DD, Mercadante VRG, Ealy AD, Johnson SE, DiLorenzo N, Lamb GC. Effects of nutrient restriction on the metabolic profile of Bos indicus-influenced and Bos taurus suckled beef cows. Animal. 2021 Mar;15(3):100166. doi: 10.1016/j.animal.2020.100166. Ganesan, S., Keating, A.F. 2020. Ovarian mitochondrial and oxidative stress proteins are altered by glyphosate exposure in mice. Toxicology and Applied Pharmacology. 402:115116. Ganesan, S., McGuire, B.C., Keating, A.F. 2020. Absence of glyphosate-induced effects on ovarian folliculogenesis and steroidogenesis. Reproductive Toxicology. 96:156-164. Garcia-Guerra A, Sala RV, Carrenho-Sala L, Baez GM, Motta JCL, Fosado M, Moreno JF, Wiltbank MC. Postovulatory treatment with GnRH on day 5 reduces pregnancy loss in recipients receiving an in vitro produced expanded blastocyst. 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Immunological detection of pregnancy: Evidence for systemic immune modulation during early pregnancy in ruminants. 2020 Theriogenology 150, 498-503. Owens CE, Daniels KM, Ealy AD, Knowlton KF, Cockrum RR. Graduate student literature review: Potential mechanisms of interaction between bacteria and the reproductive tract of dairy cattle. J Dairy Sci. 2020 Nov;103(11):10951-10960. doi: 10.3168/jds.2019-18050. Owens CE, Huffard HG, Nin-Velez AI, Duncan J, Teets CL, Daniels KM, Ealy AD, James RE, Knowlton KF, Cockrum RR. Microbiomes of various maternal body systems are predictive of calf digestive bacterial ecology. Animals 2021, 11(8), 2210; https://doi.org/10.3390/ani11082210 Padmanabhan V, Cardoso RC. Neuroendocrine, Autocrine, and Paracrine Control of Follicle-Stimulating Hormone Secretion. Molecular and Cellular Endocrinology, 500: 1-9, 2020. Pate JL. 2020. 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(2020) Luteinizing hormone regulates the phosphorylation and localization of the mitochondrial effector dynamin like 1 (DRP1) and steroidogenesis in the bovine corpus luteum. FASEB J, 32:5299-5316, PMID: 32077149 Plewes MR, Krause C, Talbott HA, Przygrodzka E, Wood JR, Cupp AS, and Davis JS. (2020) Trafficking of cholesterol from lipid droplets to mitochondria in bovine luteal cells: acute control of progesterone synthesis. FASEB J, 34: 10731-10750, PMID 32077149 Prata AB, Madureira G, Robl AJ, Ribeiro HS, Sagae M, Elias MCV, Pimenta C, Barrios J, Hartmman D, Schneider AA, Sandoval GAF, Wiltbank MC, et al. Progesterone-based timed AI protocols for Bos indicus cattle III: Comparison of protocol lengths. Theriogenology 2020; 152:29-35.  10.1016/j.theriogenology.2020.04.020 Rhoads ML. Effects of periconceptional heat stress on primiparous and multiparous daughters of Holstein dairy cows. Theriogenology. 2020 Mar 11. pii: S0093-691X(20)30181-3. doi: 10.1016/j.theriogenology.2020.03.015. Romoser, M.R., Hale, B.J., Seibert, J.T., Gall, T., Rademacher, C.J., Stalder, K.J., Baumgard, L.H., Keating, A.F., Ross J.W. 2020. Methods for reproductive tract scoring as a tool for improving sow productivity. Translational Animal Science. 4 (1): 275–284.  Sala RV, Melo LF, Motta JCL, Leffers-Neto L, Carrenho-Sala L, Fosado M, Moreno JF, Baruselli PS, Wiltbank MC, Garcia-Guerra A. 2020 Optimization of a 5-day fixed-time embryo transfer (FTET) protocol in heifers I. Manipulation of circulating progesterone through reutilization of intravaginal progesterone devices during FTET. Theriogenology, 156: 171-180.** collaborative projects as part of NE1727 Sala RV, Melo LF, Motta JCL, Leffers-Neto L, Carrenho-Sala LC, Fosado M, Moreno JF, Baruselli PS, Wiltbank MC, Garcia-Guerra A. Optimization of a 5-day fixed-time embryo transfer (FTET) protocol in heifers I. Manipulation of circulating progesterone through reutilization of intravaginal progesterone devices during FTET. 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Jr, Masello M., Prata A.B., Granados G.E., Van Amburgh M.E., Luchini D., Shaver R.D., Wiltbank M.C., Giordano J.O. 2021. Effects of feeding rumen-protected methionine pre- and postpartum on reproductive outcomes of multiparous Holstein cows. J. Dairy Sci. 104:11210-11225. https://doi.org/10.3168/jds.2021-20190 **Collaborative projects as part of NE1727 regional research project. Talbott HA, Plewes MR, Krause C, Xiaoying H, Zhang P, Rizzo WB, Wood JR, Cupp AS, and Davis JS. (2020) Formation and characterization of lipid droplets of the bovine corpus luteum. Scientific Reports 10: 11287, PMID 32647143 Timlin CL, Lynn A, Wooldridge LK, Uh K, Ealy AD, White RR, Lee K, Mercadante VRG. Physical parameters of bovine activated oocytes and zygotes as predictors of development success. Zygote. 2021 Mar 19:1-7. doi: 10.1017/S0967199421000058. Toledo M. Z., M. L. Stangaferro, R. S. Gennari, R. V. Barletta, M. M. Perez, R. Wijma, E. M. Sitko, G. Granados, M. Masello, M. E. Van Amburgh, D. Luchini, J. O. Giordano, R. D. Shaver, and M. C. Wiltbank. 2021. Effects of feeding rumen-protected methionine pre- and postpartum in multiparous Holstein cows: Lactation performance and plasma amino acid concentrations. J. Dairy Sci. 104:7583-7603. https://doi.org/10.3168/jds.2020-19021**Collaborative projects as part of NE1727 regional research project. Vanselow J, Christenson LK and Pate JL. 2020. Editorial: Regulation of Dynamic Changes and Remodeling Events During the Formation, Rescue and Regression of the Corpus Luteum. Front. Endocrinol. 11:244. doi: 10.3389/fendo.2020.00244 Wooldridge LK, Ealy AD. Interleukin-6 promotes primitive endoderm development in bovine blastocysts. BMC Dev Biol. 2021 Jan 12;21(1):3. doi: 10.1186/s12861-020-00235-z. Zhang M, Chaney HL, Current JZ and Yao J. 2021. Identification of the core promoter of ZNFO, an oocyte-specific maternal effect gene in cattle. Gene.<a href="https://doi.org/10.1016/j.gene.2021.145717"> https://doi.org/10.1016/j.gene.2021.145717</a>.  Abstracts for scientific conferences Arneson JW, Stewart JW, Harrod MK, Newberne HM, Burgess MK, Jordan JA, White RR, Ealy AD, El-Kadi SW, Rhoads RP, Rhoads ML. Impact of heat stress and glycemic state on plasma gamma-aminobutyric acid (GABA) in lactating Holstein cows. 2021 J Dairy Sci 104: Suppl 1. B. Duran, R. V. Sala, P. L. J. Monteiro, C. Gamarra, M. Fosado, E. Walleser, A. Prata, R. Gennari, J. Moreno, M. Wiltbank, A. García-Guerra. 2020.“Treatment with gonadotrophin-releasing hormone on Day 7 or 21 does not reduce pregnancy loss in dairy heifers receiving in vitro-produced embryos”. Reproduction, Fertility and Development, 2019, Vol. 32 No. 2 Page 176-177. B. T. Menichetti, A. Garcia-Guerra, J. Lakritz, W. P. Weiss, J. S. Velez, D. Merchan, G. M. Schuenemann. 2020. “Effect of timing of prepartum vaccination relative to pen change of dairy cows on lying time and serum glucose, nonesterified fatty acids, and calcium at calving”. Journal of Dairy Science 2020, Vol 103, Suppl 1, Page 175.  B. T. Menichetti, J. M. Piñeiro, A. Garcia-Guerra, A. E. Relling, W. P. Weiss, G. M. Schuenemann. 2020. “Consistency of lying time is associated with reduced serum nonesterified fatty acids of prepartum dairy heifers and cows”. Journal of Dairy Science 2020, Vol 103, Suppl 1, Page 174.  B.R. Crites, S.N. Carr, C. Hamilton, J.C. Matthews, W.R. Burris and P.J. Bridges.  2020.  Form of dietary selenium affects cholesterol biosynthesis and immune response elements in the early luteal phase bovine corpus luteum.  Special Issue, Proc. of the 53rd annual meeting of the Soc. for the Study of Reproduction. Baldwin H, Zhang M, Current J and Yao J. Expression profile of ovarian agouti-signaling protein in cattle. 47th Annual Conference of the IETS Virtual Conference. January 18-21, 2021. (Abstract). Bidne KL, Rister AL, Dodds ED, and Wood JR (2020) Maternal western diet consumption alters placental lipid composition and apolipoprotein gene expression. 53rd Annual Meeting of the Society for the Study of Reproduction, Virtual; selected for pre-doctoral platform competition Bishop C.V., Simmons W.P., Jachter S.L., De Souza V.P., Xu J., Estill C. Epidermal Growth Factor (EGF) and EGF+ Kit Ligand or Leukemia Inhibitory Factor Enhances Survival and Growth of Bovine Multilayer Follicles in 3D Matrix-free Culture System. Society for the Study of Reproduction July 2020, Virtual Poster Presentation. Bochantin KA, Keane JA, Snider AP, Kurz SG, Bergman JW, McFee RM, Cupp AS, and Wood JR (2020) Systemic inflammation in beef cows is associated with delayed puberty attainment and decreased production in small follicles. 53rd Annual Meeting of the Society for the Study of Reproduction, Virtual Bries, A., Webb, J., Vogel, B., Carrillo, C., Keating, A., Schalinske, K. 2020. Letrozole-induced polycystic ovary syndrome attenuates cystathionine-b synthase gene expression in the ovaries of female Sprague Dawley rats. Current Developments in Nutrition. 4: Suppl 2; 1244. Butler, H. Butler, E. Etcheverry, J. F. Pagani, R. Alberio, A. García-Guerra. 2020. “Treatment with gonadotrophin releasing hormone at the time of AI in beef heifers that fail to express oestrus after an estradiol-based synchronization protocol improves pregnancies per AI”. Reproduction, Fertility and Development, 2019, Vol. 32 No. 2 Page 208-208. Current J, Baldwin H, Zhang M and Yao J. Differential gene expression of bovine long noncoding RNAs in single oocytes aspirated from small and large follicles. 47th Annual Conference of the IETS Virtual Conference. January 18-21, 2021. (Abstract). D. Pereira, D. Moreno, R. Sala, L. Carrenho-Sala, M. Fosado, J. Moreno, A. García-Guerra. 2020. “SOFaaci-HEPES or holding media can be used for embryo loading without changes in pregnancies per embryo transfer nor pregnancy loss in an in vitro-produced embryo transfer program”. Reproduction, Fertility and Development, 2019, Vol. 32 No. 2 Page 178-178. de Souza V.P., Jensen J., Estill C., Bishop C. V. Increasing vitamin D levels to improve fertilization rates in cattle. Abstract Submitted for Poster Presentation to the Western Section American Society of Animal Science 2021 Meeting, October 17-20, Fort Collins, CO. E. Rojas Canadas, S. E. Battista, J. Kieffer, S. Wellert, A. García-Guerra. 2020. “Increasing gonadotrophin-releasing hormone dose at initiation of a 5-day CO-Synch protocol increases ovulatory response but not fertility in yearling beef heifers”. Reproduction, Fertility and Development, 2019, Vol. 32 No. 2 Page 206-207. Ermisch AF, Bidne KL, and Wood JR (2020) Increased adiposity and circulating glucose promote pro-inflammatory and immune cell signaling in the cumulus oocyte complex of TLR4-hyporesponsive mice fed a high fat/high sugar diet. 53rd Annual Meeting of the Society for the Study of Reproduction, Virtual González Alvarez, M.E., McGuire, B.C., Keating, A.F. 2020. Effects of zearalenone exposure on the ovarian proteome in lean and obese mice. *Accepted to Gordon Research Conference on Environmental Endocrine Disrupting Chemicals. Meeting cancelled due to COVID19. González Alvarez, M.E., McGuire, B.C., Keating, A.F. 2020. Investigating additive impact of obesity on zearalenone-induced ovotoxicity. Society of Toxicology annual meeting. *Meeting cancelled due to COVID19. González Alvarez, M.E., McGuire, B.C., Keating, A.F. 2020. Ovarian effects of perfluorooctanoic acid exposure in lean and obese mice. Society for the Study of Reproduction annual meeting. Goulet M, Hutchings D, Elder D, Tsang PCW.  2021. Regulation of Cellular Communication Network Factor 1 by Ras Homolog Family Member A in Bovine Luteal Cells. 55th Annual Meeting of the Society for the Study of Reproduction, Saint Louis, Missouri (held Dec 15 – Dec 18 due to COVID-19; hybrid mode of presentation). Granados G. E., M.M Pérez, and J. O. Giordano*. 2020. Pattern of Behavioral, Physiological, and Performance Parameters before Insemination in Dairy Cows that Become Pregnant or Not to First Service.  J. Dairy Sci. Volume 103. E-Supplement 1. Gurung S, Koganti PP, Schalich KS, and Selvaraj V. Absence of evidence for physiological fetal microchimerism across the epitheliochorial bovine placental interface Hessock, E.A., M.F. Smith, T.W. Geary, K.G. Pohler, J.A. Green, S.K. Behura, A.K. Stokes, and S.E. Moorey. Transcriptome profiles of in vivo matured bovine cumulus-oocyte complexes reveal unique gene expression between the oocyte and cumulus cells. 54th Annual Meeting of the Society for the Study of Reproduction (SSR), St Louis, MO. Horn, E.J., C.C. Read, J.L. Edwards, F.N. Schrick, J.D. Rhinehart, R.R. Payton, S.R. Campagna, H.F. Castro, J.L. Klabnik, and S.E. Moorey. Pre-ovulatory follicular fluid and serum metabolome profiles in beef cows with thin, moderate, and obese body condition. 54th Annual Meeting of the Society for the Study of Reproduction (SSR), St Louis, MO. K. Nickles, A. E. Relling, A. Garcia-Guerra, F. L. Fluharty, A. J. Parker. 2020. “Cows housed in muddy environmental conditions during late gestation have lesser body weight and body condition score compared with cows housed on wood chips”. Journal of Animal Science, Volume 98, Issue Supplement 4, November 2020, Page 48. Keane JA, Nafziger SR, Bergman JW, Kurz SG, Snider AP, Bochantin KA, Wood JR, Cushman RA, Summers AF, and Cupp AS (2020) Environmental parameters may increase likelihood of beef heifers classified with earlier or later pubertal attainment. 53rd Annual Meeting of the Society for the Study of Reproduction, Virtual Lupicka, M., JL Pate. 2020. MiR-1246 is the most abundant miRNA in luteal extracellular vesicles and it regulates T cell transcripts associated with their activation. Society for the Study of Reproduction (virtual poster session). L. Carrenho-Sala, M. Fosado, R. Sala, E. Peralta, D. Pereira, D. Moreno, J. Moreno, A. García-Guerra. 2020. “Synchronization of follicle wave emergence prior to superstimulation with purified FSH for ovum pickup affects blastocyst rate in pregnant Holstein heifers”. Reproduction, Fertility and Development, 2019, Vol. 32 No. 2 Page 215-216. Masello M., M. M. Perez, G. E. Granados, M. L. Stangaferro, B. Ceglowski, M. J. Thomas, and J. O. Giordano*. 2020. Reduced age at first calving improved the economic performance of replacement dairy heifers.  J. Dairy Sci. Volume 103. E-Supplement 1. Masello M., Y. Ren, D. Erickson, and J. O. Giordano*. 2020. An electronically-controlled intravaginal hormone delivery device successfully induced luteal regression in dairy cattle.  J. Dairy Sci. Volume 103. E-Supplement 1. Mathew, D.J., H.L. Chaney, L.F. Grose, G. Charpigny, S.K. Behura, I.M. Sheldon, J.G. Cronin, T.E. Spencer, and P. Lonergan. Effect of galectin-1 on bovine endometrial epithelial and fibroblast cell transcriptomes in 3D cell culture. 54th Annual Meeting of the Society for the Study of Reproduction (SSR), St Louis, MO. Maucieri AM, Townson DH (2021) Evaluating the impact of the hexosamine biosynthesis pathway and O-GlcNAcylation on glucose metabolism in bovine granulosa cells. Society for the Study of Reproduction Annual Meeting. St. Louis, MO. McGuire, B.C., Ganesan, S., Keating, A.F. 2020. Impact of Glyphosate exposure on the neonatal ovarian proteome. *Accepted to Gordon Research Conference on Environmental Endocrine Disrupting Chemicals. Meeting cancelled due to COVID19. McGuire, B.C., González Alvarez, M.E., Keating, A.F. 2020. Determining impacts of glyphosate exposure on ovarian function in lean and obese mice. Society of Toxicology annual meeting. *Meeting cancelled due to COVID19. McGuire, B.C., González Alvarez, M.E., Keating, A.F. 2020. Exposure to glyphosate alters the ovarian proteome with differential effects in lean and obese mice. Society for the Study of Reproduction annual meeting. Mezera MA, Hughes CHK, Pate JL, Wiltbank MC. 2020. The impact of pregnancy on the transcriptome of the bovine CL: insights from combining two independent experiments.  Society for the Study of Reproduction (virtual poster session). Novbotova, G., Ganesan, S., McGuire, B.C., Keating, A.F. 2020. Maternal and ovarian impacts of chronic glyphosate exposure. Society for the Study of Reproduction annual meeting. Oliver, M.A., K.D. Peterson, S. Bhandari and D.J. Mathew. Effect of progesterone on bovine endometrial cell expression of select transporters and growth factors. American Dairy Science Association 2021 Annual meeting. Virtual. Oliver, M.A., K.D. Peterson, S. Bhandari, R.R. Payton, L.G. Schneider, J.L. Edwards, and D.J. Mathew. The effect of endometrial cell conditioned media on in vitro cultured bovine embryo development. 54th Annual Meeting of the Society for the Study of Reproduction (SSR), St Louis, MO. Peterson, K.D., M.A. Oliver, S. Bhandari, S.E. Moorey, and D.J. Mathew. 2021. The effect of small and large ovarian follicle fluid on oviduct cell gene expression in cattle. American Dairy Science Association 2021 Annual meeting. Virtual. R. Sala, L. Carrenho-Sala, V. Absalon-Medina, A. Lopez, M. Fosado, J. Moreno, M. Wiltbank, A. García-Guerra. 2020. “Optimization of a five-day fixed-time embryo transfer program in dairy heifers: Use of gonadotrophin-releasing hormone at initiation of the protocol”. Reproduction, Fertility and Development, 2019, Vol. 32 No. 2 Page 179-179. Read, C.C., J.L. Edwards, F.N. Schrick, J.D. Rhinehart, R.R. Payton, S.R. Campagna, H.F. Castro, J.L. Klabnik, and S.E. Moorey. Influence of pre-ovulatory follicle size on the follicular fluid metabolome in lactating beef cows. 54th Annual Meeting of the Society for the Study of Reproduction (SSR), St Louis, MO. Rishi, J.K., Roach, C.M., Baumgard, L.H., Ross, J.W., Keating, A.F. 2020. Impact of heat stress on ovarian chemical biotransformation enzyme abundance. Society of Toxicology annual meeting. *Meeting cancelled due to COVID19. Roach, C.M., Clark, K.L., Keating, A.F. 2020. Examining the effects of obesity on circulating prolactin and the ovarian proteome in hyperphagia-induced obese mice. Society for the Study of Reproduction annual meeting. Roach, C.M., Mayorga, E.J., Ross, J.W., Baumgard, L.H., Keating, A.F. 2020. Zearalenone alters physiological function in heat-stressed prepubertal gilts. *Accepted to Gordon Research Conference on Environmental Endocrine Disrupting Chemicals. Meeting cancelled due to COVID19. Roach, C.M., Mayorga, E.J., Ross, J.W., Baumgard, L.H., Keating, A.F. 2020. Heat stress alters reproductive phenotypic impacts of Zearalenone in pre-pubertal gilts. MidWest American Society of Animal Science meeting. S. R. Wellert, S. E. Battista, J. Kieffer, R. N. Lurch, A. García-Guerra. 2020. “Comparison of different Doppler ultrasound settings for pregnancy diagnosis based on corpus luteum perfusion at 21 days after AI in beef cattle”. Reproduction, Fertility and Development, 2019, Vol. 32 No. 2 Page 211-211. S.N. Carr, B.R. Crites, Y. Jia, C. Hamilton, J.C. Matthews and P.J. Bridges.  2020.  Form of supplemental selenium affects steroidogenesis by the early luteal phase corpus luteum in beef cattle.  Special Issue, Proc. of the 53rd annual meeting of the Soc. for the Study of Reproduction. Schalich KM, Koganti PP, Herren A, Castillo J, Reiff O, Cheong S, and Selvaraj V. (2020). Enhanced resolution and direct profiling of bovine uterine fluid secretions through data-independent acquisition (DIA) in mass-spectrometry based proteomics. Schilkowsky E. M., G. E. Granados, E M. Sitko, M. Masello, M. M. Perez, and J.O. Giordano*. 2020. Evaluation and characterization of estrus alerts from an ear-attached accelerometer-based automated estrus detection system.  J. Dairy Sci. Volume 103. E-Supplement 1. Sitko E. M., M.M. Pérez, G.E. Granados, M. Masello, and J.O. Giordano*. 2020. Age at first calving affected reproductive performance of primiparous cows managed with a program that favored timed artificial insemination. J. Dairy Sci. Volume 103. E-Supplement 1. Sitko E.M., M.M. Pérez, G.E. Granados, M. Masello, and J.O. Giordano*. 2020. Association between age at first calving and reproductive performance of primiparous cows managed with a program that favored insemination at detected estrus. J. Dairy Sci. Volume 103. E-Supplement 1. Snider AP, Gomes RS, Summers AF, Abedal-Majed MA, Tenley SC, Wood JR, Davis JS, and Cupp AS (2020) Lipids involved in pro- and anti-inflammatory responses are altered in follicular fluid and plasma of cows administered a low-dose-FSH protocol and may be used as markers of ovulation in beef cows. 53rd Annual Meeting of the Society for the Study of Reproduction, Virtual; selected for post-doctoral poster competition Stangaferro M. L., J. O. Giordano*, M. Z. Toledo, and M. C. Wiltbank. 2020. Prediction of pregnancy probability at first service in multiparous dairy cows based on early lactation data.  J. Dairy Sci. Volume 103. E-Supplement 1. **Collaborative projects as part of NE1727 regional research project. Stewart J, Newberne H, Arneson A, Harrod M, Negron-Perez V, Haines H, Jordan J, White R, Ealy A, El-Kadi S, Rhoads R and Rhoads M. Glucose infusion during heat stress restores normoglycemia but does not improve milk production. 2020. J. Dairy Sci. Vol. 103, Suppl. 1. 148. Stewart JW, Newberne HM, Harrod MK, Arneson AG, Negron-Perez VM, White RR, Ealy AD, El-Kadi SW, Rhoads RP, Rhoads ML. Heat stress and glycemic state alter milk production and composition in Holstein dairy cows. 2021 American Dairy Science Association Annual Meeting, 2021 J Dairy Sci 104: Suppl 1. Studer, J.M., Kiefer, Z.E., Keating, A.F., Baumgard, L.H., Olsen, K.M., Rambo, Z., Wilson, M.E., Rapp, C., Ross, J.W. 2020. Influence of dietary manganese supplementation on transcript and protein abundance in the corpus luteum of swine. Society for the Study of Reproduction annual meeting. Toledo M. Z., M. L. Stangaferro, R. S. Gennari, P. L. J. Monteiro Jr., M. M. Perez, M. Masello, M. E. Van Amburgh, D. Luchini, R. D. Shaver, J. O. Giordano, M. C. Wiltbank. 2020. Effect of feeding rumen-protected methionine pre- and postpartum on health, herd exit dynamics and time to pregnancy in multiparous dairy cows.  J. Dairy Sci. Volume 103. E-Supplement 1. **Collaborative projects as part of NE1727 regional research project. Torres-Gonzalez E, Arbeithuber B, Hester J, Cremona M, Stoler N, Higgins B, Anthony K, Chiaromonte F, Diaz F, and Makova K.  Duplex sequencing uncovers age-related increase in the frequency of de novo indels in mouse mitochondrial DNA. Society for Molecular Biology and Evolution Annual Meeting. Quebec City, CA, June 28th- July 2nd, 2020. Zhang M, Baldwin H, Current J and Yao J. Characterization of the promoter region of ZNFO, an oocyte-specific gene in cattle. 47th Annual Conference of the IETS Virtual Conference. January 18-21, 2021. (Abstract). Theses/Dissertations Alicia Arneson (2021) Evaluating the impact of heat stress and altered glycemic state on plasma ɣ-Aminobutyric Acid (GABA) in lactating Holstein cows (MS Thesis).  Mary-Kathryn Harrod-Byrd (2021) Serial measurements of circulating glucose and luteinizing hormone concentrations in lactating dairy cattle (MS Thesis). Constantino J. Preantral Follicle Numbers and Size in Heifers Carrying the Bovine High Fecundity Allele Trio. OSU Undergraduate Honors thesis. Cooper L. Circulating AMH, antral follicle count and ovulation rate after unilateral ovariectomy in cattle: influence of the bovine fecundity allele Trio. OSU Undergraduate Honors thesis. Cunningham, Devin (2021) Effects of an extruded flaxseed supplement on plasma haptoglobin, non-esterified fatty acids, beta-hydroxybutyrate, and complete blood count in transition dairy cows and heifers. (MS Thesis). Gurung S (2021). Bovine synepitheliochorial placentation is a barrier to fetal microchimerism. (MS Thesis) Michelle L. Kott (2021). Effects of mid-gestational L-citrulline supplementation to twin-bearing ewes on umbilical blood flow, placental development, and lamb production traits (MS Thesis).  Maucieri AM, Townson DH (2020) Characterization and manipulation of O-GlcNAcylation in granulosa cells of bovine ovarian antral follicles. UVM MS Thesis Mary Ali Oliver. The effect of endometrial cell conditioned media on in vitro cultured bovine embryo development. (MS Thesis; DJM MS student) Schalich KM (2021). Analysis of aspects of the uterine environment and of early mammary secretions critical for embryonic, fetal, and neonatal development in cattle. (PhD Thesis) Zackary K. Seekford (2020). Interleukin-6 Supplementation Improves Post-Transfer Embryonic and Early Fetal Development of In Vitro Produced Bovine Embryos (MS Thesis). Planned Master’s Thesis: “Epidermal Growth Factor and Leukemia Inhibitory Factor Supplementation in Matrix-free, Three-Dimensional Bovine Follicle Culture.” Wilson Simmons, M.S. 09/2021. Major Professor C. Bishop.  Sarah Singleton. Novel postpartum reproductive phenotypes in beef cattle. (Undergraduate Chancellor’s Honor’s Thesis; SEM undergraduate student) Master’s Thesis: “Increasing vitamin D levels to improve fertilization rates in cattle.” Vanessa Peixoto de Souza, M.S., D.V.M. 06/14/2021. Major Professors C. Estill and C. Bishop. Lydia K. Wooldridge (2020). Supplementing Bovine Embryo Culture Media to Improve the Production and Quality of In Vitro Produced Bovine Embryos (PhD Dissertation).  Active collaboration within the group OSU and WI: evaluated and currently working on projects to evaluate the effect of dose of GnRH at initiation of a 5-day synchronization protocol for FTET.  NY and WI: working collaboratively on projects to determine physiological differences among cows of different genetic merit for fertility. PA and WV: Samples shared for analysis of molecular regulators of luteal function.  In addition to Hatch Multistate Funds, these studies were supported by:  USDA NIFA/AFRI  Oregon State University Animal and Rangeland Sciences Department Funds  Oregon State University Agricultural Research Foundation  Oregon Beef Council  OARDC SEEDS Early career investigator – The Ohio State University  CFAES Honors Will C. Hauk Endowment Grant – Undergraduate student - The Ohio State University  National Institute of Environmental Health Science  Iowa State University Nutritional Sciences Council Martin Fund  National Pork Board  Texas A&M University - Triads for Transformation  American Simmental Association  University of Tennessee AgResearch Instrumentation Grants program  NIH-R21  Binational Agriculture Research and Development (BARD)  Office of Research and Economic Development, Univ Nebraska-Lincoln Bill and Melinda Gates Foundation NIH/NIGMS-sponsored Ruth L. Kirschstein National Research Service Award (NRSA) Institutional Research Training (T32)

Impact Statements

Low pregnancy rates in beef and dairy cows result in increased costs to producers associated with increased rebreeding or culling of animals, and reduced likelihood of recouping heifer development costs. Reduced fertility is also a major cause of reduced milk (dairy) and meat (beef) production which impacts the food supply. The focus of the NE1727 multistate project “Influence of ovary, uterus, and embryo on pregnancy success in ruminants”, is to understand how altered ovarian function, impaired oocyte quality, and disruptions of conceptus-uterine interactions contribute to infertility, and devise management strategies that will overcome these factors in order to optimize the chances that animals successfully achieve a pregnancy. To achieve this goal, the following three objectives were performed: (1) Determine the impact of altered ovarian function on reproductive performance; (2) Identify alterations in embryo development and uterine and CL function associated with declining pregnancy establishment; and (3) Identify changes in genetics and reproductive management that lead to improved pregnancy rates. As investigators in NE1727, we have used our collective expertise to broaden the current understanding of the mechanisms by which internal and external forces modify these important physiological processes. Furthermore, this information about core biological processes was used to rationally develop new and innovative tools to increase cyclicity and breeding efficiency, and reduce early embryonic failures. The outcome of these accomplishments was improving reproductive efficiency of both dairy and beef cattle operations in the United States.

Date of Annual Report: 08/05/2022

Report Information

Annual Meeting Dates: 05/17/2022 - 05/18/2022
Period the Report Covers: 10/01/2021 - 09/30/2022

Participants

Participants Attachment:

NE1727 Annual Report from Meeting. JG 08032022.docx

Brief Summary of Minutes

Minutes Attachment:

NE1727 Minutes. 2022 Virtual Meeting.docx

Accomplishments

Demonstrated that a targeted reproductive management program for first service that combined insemination at detected estrus after automated estrus alerts and timed AI led to similar reproductive performance than an all-TAI program. Also, demonstrated that cows with at least one automated estrus alert during the voluntary waiting period had improved reproductive performance. Thus, automated estrus detection systems can be used to identify cows with different expected level of reproductive success for targeted management. Demonstrated in an experiment with multiple farms that lactating dairy cows of superior genetic merit for fertility have better reproductive performance than cows of inferior genetic for fertility regardless of the method of submission to insemination.   Evaluated and demonstrated the reproductive and economic value of targeted reproductive management strategies that used genomically enhanced predictions for fertility to identify subgroups of cows for targeted management.   Demonstrate that cows of inferior genetic merit for fertility as determined by genomic daughter pregnancy rate are more likely to have atypical estrous cycles due to a multitude of physiological failures including premature luteal regression, extended luteal phases, anestrus, anovulation, and development of follicular cysts.   Determined the effect of form of Se on the transcriptome of the early luteal phase corpus luteum (CL) with the goal of elucidating form of Se-regulated processes affecting luteal steroidogenesis and function.   Determined effects of form of Se on the development of the bovine conceptus and the endometrium using targeted qPCR on day 17 of gestation, the time of maternal recognition of pregnancy (MRP).   Determined the effect of form of Se on the expression of mRNA encoding selenoproteins in the corpus luteum (CL), and whether the previously reported MIX-induced increase in P4 is the result of increased luteal expression of key steroidogenic transcripts. Overall, we observed that the form of Se provided to cows is reported to affect the expression of mRNA encoding several selenoproteins in the CL, and that the form of Se-induced effects on luteal production of P4 appears to be the result of changes in cholesterol uptake, rather than a direct effect on the expression of steroidogenic enzymes within the CL.   Generated evidence to support the hypothesis of a physiological mechanism, involving increased uterine blood flow, is responsible for maintenance of the CL during the second month of pregnancy in cattle. The 5-day CO-Synch is an extensively used TAI protocol; however, it requires multiple administrations of prostaglandin F2α (PGF). We conducted a study to evaluate the effect of increasing progesterone (P4) device insertion from 5 to 6 days during a TAI protocol on ovarian dynamics and fertility in beef cows. Cows in the 6-day group had a larger maximum preovulatory follicle diameter and a greater CL volume seven days after ovulation than cows in the 5-day group. Furthermore, cows assigned to the 6-day group had greater estrus expression and pregnancy rate than cows in the 5-day group. Demonstrated that increasing the length of P4 during a prolonged proestrus CO-Synch protocol by one day improved fertility to ET by reducing pregnancy loss. Demonstrated that pregnancies per AI (P/AI) on day 35 after TAI were greater for cows presynchronized with a progesterone device and prostaglandin compared to cows presynchronized with prostaglandin alone or cows that did not undergo presynchronization. Determined the effects of prostaglandin F2alpha (PGF2α) on CCN1 expression in ovarian adenocarcinoma 8 (OVCAR8) cells. Determined the effects of estradiol (E2) on CCN1 expression in ovarian tumor granulosa (KGN) cells. Determined the effects of a protein kinase A (PKA) inhibitor (PKI), and peptide 17, an inhibitor of yes-associated protein/transcriptional enhanced associate domain (YAP/TEAD) protein-protein interaction, on CCN1 expression in steroidogenic luteal cells obtained from the 4-day-old bovine corpus luteum. Determined the effects of the phytoestrogen, Daidzein, on CCN1 and VEGFA expression in steroidogenic luteal cells obtained from the 4-day-old bovine corpus luteum. Determined the effects of angiotensin II (Ang II) on CCN1 expression in steroidogenic luteal cells obtained from the 4-day-old bovine corpus luteum. Localized expression of ZIP9 to mural and cumulus granulosa cells in mouse and bovine Identified ZIP9 localization to the mitotic spindle in granulosa cells. Provided evidence that pregnancy in dairy cattle is associated with altered expression of molecules related to immune tolerance in peripheral blood leukocytes. Demonstrated that AMH levels were not different in females by pregnancy status either 1 week before, or on day of AI. Similarly, serum levels of inhibin A were not significantly different by pregnancy status when analyzed pre-breeding, however on the day of AI there was a weak tendency for females who did not become pregnant to have lower levels of inhibin A. Identified 29 follicular fluid metabolites with a positive relationship with pre-ovulatory follicle diameter and 22 metabolites that had a positive relationship with serum estradiol concentration at the time of GnRH injection to induce ovulation.   Determined that there is a positive relationship between serum estradiol concentration at the time of GnRH injection to induce ovulation and oocyte ATP level ~18 hours later.   Identified 1,387 and 2,765 mRNA transcripts with unique expression and 2,553 and 1,745 mRNA transcripts with upregulated expression in in vivo matured bovine oocytes or cumulus cells, respectively. This improves our understanding of oocyte and cumulus function during in vivo maturation.   Generated new knowledge highlighting that heat-induced increases in body temperature in lactating dairy cows differentially impacts the cumulus and granulosa cell transcriptome of the periovulatory follicle.   Characterized expression of galectin molecules, a family of lectins with essential functions during establishment of pregnancy in rodents, in bovine and ovine endometrium and conceptus tissues.   Tested and found that bovine galectin-1 has immunomodulatory effects within bovine endometrium related to maternal-conceptus immune tolerance.   Identified progesterone stimulated bovine endometrial epithelial and stroma fibroblast cell specific mRNAs (better understand uterine support of early embryonic development).   Tested and found an advantage of progesterone stimulated bovine endometrial epithelial and stroma fibroblast cell conditioned media on in vitro produced (IVP) bovine blastocyst development.   Identified 21 serum metabolites, from blood collected in the pre-ovulatory time period, that differed among beef cows with thin, moderate, and obese body condition scores. This helps understand the impact of body condition on fertility.   Showed that there is a positive relationship of rectal temperature at fixed timed artificial insemination on pregnancy outcomes in beef cattle during a regular breeding season where pregnancy outcomes are expected to exceed 50%.   Demonstrated that Chromium propionate supplementation during the early postpartum period (days 20-44 in milk) tends to increase the number of 6-9 mm ovarian follicles in multiparous Holstein cows. It also increases the mean diameter of follicles in this category.   Reported that early lactation dairy cattle fed a chromium propionate supplement have a greater ratio of plasma progesterone to corpus luteum volume.   Examined the role of cytokines in the initial development of the inner cell mass with blastocyst stage embryos. This served to establish the lowest effective concentration and determined the recombinant human IL6 concentration that is biologically active in bovine embryos.   Developed a CRISPR-Cas9 gene editing strategy that blocks expression of IL6 signal transducer, the predominant signaling component of the IL6 family receptor complex, reduces bovine blastocyst development.   Evaluated the effects of IL6 cytokine family members on bovine oocyte maturation.   Demonstrated that Chromium propionate supplementation during early lactation may protect against a resurgence in uterine inflammation as cows at 44 days in milk experienced no change in uterine polymorphonuclear leukocyte percent when consuming the supplement while control cows exhibited an increase in uterine polymorphonuclear leukocyte percent. Improved understanding of the mechanisms involved in selection of a single dominant follicle in cattle. This will be key for the rational design of new reproductive management strategies and reproductive biotechnologies. Discovered the mechanism that produces increased ovulation rate in carriers of the Trio allele.  This mechanism may also underlie double ovulation in dairy cattle and in other species. This should allow us to manipulate the ovulation rate in cattle and perhaps other species. We have now determined how to practically utilize Trio to reliably produce bilateral twins in beef cattle. This may be of great practical value to beef cattle producers in the future. Demonstrated that the corpus luteum is maintained during early pregnancy by suppression of pulses of prostaglandin F2a by interferon-tau, based on our measurements of PGFM surges. We have also discovered that there is a second period of corpus luteum maintenance after Day 25 when interferon-tau secretion ends and PGF pulses are initiated. The resolution of the mechanisms that allow maintenance of the corpus luteum during this second period will be key for reducing pregnancy loss in cattle. Demonstrated that increasing circulating progesterone can reduce pregnancy loss in lactating dairy cows and in recipients of in-vitro produced embryos. This provides an easy method to reduce pregnancy loss by treatment with either GnRH or hCG. Identified profiles of gut microbial families that are associated with increased ovarian inflammation and reduced reproductive performance in beef cows.   Determined the mechanistic effects of inflammation and oxidative stress on bovine somatic cell steroidogenesis, oocyte maturation and pre-implantation embryo development.

Publications

Crites, B.R., S.N. Carr, J.C. Matthews and P.J. Bridges. 2022. Form of dietary selenium affects mRNA encoding cholesterol biosynthesis and immune response elements in the early luteal phase bovine corpus luteum. Journal of Animal Science. In Press. Crites, B.R., S.N. Carr, L.H. Anderson, J.C. Matthews and P.J. Bridges. 2022. Form of dietary selenium affects mRNA encoding interferon-stimulated and progesterone-induced genes in the bovine endometrium and conceptus length at maternal recognition of pregnancy. Journal of Animal Science. In Press. Carr, S.N., B. Crites, J.L. Pate, C.H.K. Hughes, J.C. Matthews and P.J. Bridges. 2022. Form of supplemental selenium affects the expression of mRNA transcripts encoding selenoproteins, and proteins regulating cholesterol uptake, in the corpus luteum of grazing beef cows. Animals. 12:313.   Menichetti B.T., Garcia-Guerra A, Lakritz J., Weiss, W. P. Velez J. S., Bothe H., Merchan D., Schuenemann G. M. 2021. Effect of timing of prepartum vaccination relative to pen change with an acidogenic diet on lying time and metabolic profile in Holstein dairy cows. Journal of Dairy Science, 104 (10): 11059-11071.   Menichetti B.T., Garcia-Guerra A, Lakritz J., Weiss, W. P. Velez J. S., Bothe H., Merchan D., Schuenemann G. M. 2021. Effects of prepartum vaccination timing relative to pen change with an acidogenic diet on serum and colostrum immunoglobulins in Holstein dairy cows. Journal of Dairy Science, 104 (10): 11072-11081.   Duran B. J., F. L. V. Pinaffi, J. C. L. Motta, C. Hayden, A. E. Crist, C. Rykaczewski, S. Wellert, E. Rojas-Canadas, M. L. Mussard, A. García-Guerra. 2021. “Corpus luteum regression after induced pregnancy loss in cattle is preceded by a reduction in uterine artery vascular perfusion”. Abstract P136. Proceedings of the 54th Annual Meeting of the Society for the Study of Reproduction 2021, December 13-18, St Louis, MO.   García-Guerra A., L. Cooper, B. W. Kirkpatrick, M. C. Wiltbank. 2021. “Circulating AMH, antral follicle count, and ovulation rate after unilateral ovariectomy in cattle: influence of the bovine fecundity allele Trio”. Abstract P150. Proceedings of the 54th Annual Meeting of the Society for the Study of Reproduction 2021, December 13-18, St Louis, MO.   Crist A., J. C. L. Motta, C. Hayden, B. Duran, M. Mussard, A. Garcia-Guerra. 2021. “Ovarian dynamics and fertility in beef cows administered a 5-day or 6-day CO-Synch protocol for timed artificial insemination”. Journal of Animal Science, Volume 99, Issue Supplement 4, November 2021, Page XX.   Motta J. C. L., R. V. Sala, V. A. Absalón-Medina, V. C. Fricke, M. Dominguez, D. C. Pereira, C. Hayden, E. R. Canadas, B. J. Duran, J. F. Moreno, A. García-Guerra. 2021. “Influence of length of porcine follicle-stimulating hormone (p-FSH) treatment before ovum pickup on ovarian response and in vitro embryo production in Holstein heifers”. Reproduction, Fertility and Development, 2021, Vol. 33 No. 2, Page 170-171.   Canadas E. R., B. Duran, G. Machado, A. Nall, S. E. Battista, M. Mussard, P. S. Baruselli, A. García-Guerra. 2021. “Presynchronization and reutilization of progesterone devices during a 6-day CO-Synch protocol for fixed-time artificial insemination in beef heifers”. Reproduction, Fertility and Development, 2021, Vol. 33 No. 2, Page 165-166.   Brochado C., B. J. Duran, J. C. L. Motta, J. D. Kieffer, A. Pinczak, A. Menchaca, A. García-Guerra. 2021. “Simplification of the follicle-stimulating hormone protocol for superovulation of the first follicular wave in sheep”. Reproduction, Fertility and Development, 2021, Vol. 33 No. 2, Page 182.   Duran B. J., J. C. Lemos-Motta, E. Rojas-Canadas, C. Hayden, C. Rykaczewski, A. Nall, M. L. Mussard, A. García-Guerra. 2021. “Timing of luteolysis and conceptus expulsion after induced pregnancy loss at three different timepoints after maternal recognition of pregnancy in cattle”. Reproduction, Fertility and Development, 2021, Vol. 33 No. 2, Page 131.   Hayden C., R. V. Sala, D. C. Pereira, L. C. Carrenho-Sala, M. Fosado, D. Moreno, J. F. Moreno, A. García-Guerra. 2021. “Effect of follicle-stimulating hormone dose and circulating progesterone before ovum pickup and in vitro embryo production in pregnant Holstein heifers”. Reproduction, Fertility and Development, 2021, Vol. 33 No. 2, Page 143.   Goulet MR, Hutchings D, Donahue J, Elder D, Tsang PCW. 2022. Regulation of cellular communication network factor 1 by Ras homolog family member A in bovine steroidogenic luteal cells. J Anim Sci, in press Hughes CHK, Rogus A, Inskeep EK, Pate JL. 2021. NR5A2 and potential regulatory miRNAs in the bovine CL during early pregnancy. Reproduction <a href="https://doi.org/10.1530/REP-20-0009">https://doi.org/10.1530/REP-20-0009</a> Carr SN, Crites BR, Pate JL, Hughes CHK, Matthews JC, Bridges PJ. 2022. Form of Supplemental Selenium Affects the Expression of mRNA Transcripts Encoding Selenoproteins, and Proteins Regulating Cholesterol Uptake, in the Corpus Luteum of Grazing Beef Cows. Animals2022,12,313. https:// doi.org/10.3390/ani12030313 Hughes CHK, Mezera M, Wiltbank M, Pate JL. 2022. Insights from two independent transcriptomic studies of the bovine corpus luteum of pregnancy. J Anim Sci. in press Bishop C, Selvaraj V, Townson D, Pate J, Wiltbank M. 2022. History, insights and future perspectives on studies into luteal function in cattle. J Anim Sci in press Oh, J., Harper, M. T., Melgar, A., Raisanen, S., Chen, X., Nedelkov, K., Fetter, M., Ott, T., Wall, E. H., & Hristov, A. N. (2021). Dietary supplementation with rumen-protected capsicum during the transition period improves the metabolic status of dairy cows. Journal of Dairy Science, 104(11), 12 pages. DOI: https://doi.org/10.3168/jds.2020-19892 Fetter, M. E., Cunningham, D. M., Gambonini, F., Raisanen, S. E., Ott, T. L., & Hristov, A. N. (2021). Short communication: Postpartum plasma amino acid concentration in primi- and multiparous Holstein cows. Animal Feed Science and Technology, 281, 8 pages. DOI: <a href="https://doi.org/10.1016/j.anifeedsci.2021.115101">https://doi.org/10.1016/j.anifeedsci.2021.115101</a> Ott, TL. An interview with Dr. Fuller Warren Bazer. (2021). Biology of Reproduction 104(2), 248-251. Jachter SL, Simmons WP, Estill C, Xu J, Bishop CV. Matrix-free three-dimensional culture of bovine secondary follicles to antral stage: Impact of media formulation and epidermal growth factor (EGF). Theriogenology. 2022 Mar 15;181:89-94. doi: 10.1016/j.theriogenology.2022.01.013. Epub 2022 Jan 12. PMID: 35066368; PMCID: PMC8871473. de Souza V.P., Jensen J., Estill C., Bishop C. V. Increasing vitamin D levels to improve fertilization rates in cattle. Manuscript Accepted Journal of Animal Sciences, 05/5/2022.  Bishop, C.V. Selvaraj, V. Townson, D.H., Pate, J.L., Wiltbank, M.C. History, Insights, and Future Perspectives on Studies into Luteal Function in Cattle. Manuscript Accepted Journal of Animal Sciences, 04/12/2022. Ault-Seay T. B., R. R. Payton, S. E. Moorey, K. G. Pohler, F. N. Schrick, E. A. Shepherd, B. H. Voy, K. H. Lamour, D. J. Mathew, P. R. Myer and K. J. McLean. 2022. Bovine uterine endometrial response to lipopolysaccharide in the luteal and follicular phases between the contralateral and ipsilateral horns. Frontiers in Animal Science. In Review.   Ault-Seay T. B., K. J. Brandt, M. T. Henniger, R. R. Payton, D. J. Mathew, S. E. Moorey, F. N. Schrick, K. G. Pohler, T. P. L. Smith, J. D. Rhinehart, L. G. Schneider, K. J. McLean and P. R. Myer. 2022. Bacterial communities of the uterus and rumen during heifer development with protein supplementation. Frontiers in Animal Science. In Review.   Watson, E.M., K.A. Kurth, D.L. Metts, S.E. Moorey, B.F. Miller, and L.I. Muller. 2022. Evaluating the efficacy of large-scale, non-invasive fecal sampling for pregnancy detection in elk. Journal of Wildlife Diseases. In Review.   Walker B.N., J.L. Nix, C. Wilson, M.A. Marrella, S.L. Speckhart, L. Wooldridge, C. Yen, J.S. Bodmer, L.T. Kirkpatrick, S.E. Moorey, D.E. Gerrard, A. Ealy, and F.H. Biase. 2022. Tight gene co-expression between oocyte and surrounding cumulus is critical for oocyte developmental competence. Biology of Reproduction In Review.   Klabnik J., L.K. Christenson, S. Gunewardena, K.G. Pohler, L.A. Rispoli, R.R. Payton, S.E. Moorey, F.N. Schrick and J.L. Edwards. 2022. Heat-induced increases in body temperature in lactating dairy cows: impact on the cumulus and granulosa cell transcriptome of the periovulatory follicle. 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Edwards, F.N. Schrick, J.D. Rhinehart, R.R. Payton, S.R. Campagna, H.F. Castro, J.L. Klabnik, and S.E. Moorey. 2022. Pre-ovulatory serum estradiol concentration is positively associated with oocyte ATP and follicular fluid metabolite abundance in lactating beef cattle. Journal of Animal Science. Accepted, In Press.   Mathew D. J., K. D. Peterson, L. K. Senn, M. A. Oliver and A. D. Ealy. 2022. Review: Ruminant Conceptus-Maternal Interactions: Interferon Tau and Beyond. Journal of Animal Science. doi: 10.1093/jas/skac123. Accepted, In Press. Embargo until July, 2022.   Read, C.C., J.L. Edwards, F.N. Schrick, J.D. Rhinehart, R.R. Payton, S.R. Campagna, H.F. Castro, J.L. Klabnik, E.J. Horn, and S.E. Moorey. Correlation between pre-ovulatory follicle diameter and follicular fluid metabolome profiles in lactating beef cows. Metabolites. 2021. 11(9):623. doi: 10.3390/metabo11090623.   Read, C.C., S. Bhandari, and S.E. Moorey. 2021. Concurrent measurement of mitochondrial DNA copy number and ATP concentration in single bovine oocytes. Methods and Protocols. 4(4):88. doi: 10.3390/mps4040088.   Chaney H. L., L. F. Grose, J. M. LaBarbara, A. W. Sirk, A. M. Blancke, J. M. Sánchez, C. Passaro, P. Lonergan, and D. J. Mathew. 2021. Galectin-1 induces gene and protein expression related to maternal-conceptus immune tolerance in bovine endometrium. Biology of Reproduction. 106(3):487-502. doi: 10.1093/biolre/ioab215 Moorey, S.E., J.M. Monnig, M.F. Smith, M.S. Ortega, J.A. Green, K.G. Pohler, G.A. Bridges, S.K. Behura, T.W. Geary. Differential transcript profiles in cumulus-oocyte complexes originating from pre-ovulatory follicles of varied physiological maturity in beef cows. Genes. 2021. 12(6), 893. doi: 10.3390/genes12060893.   Bisinotto RS, Ribeiro ES, Greco LF, Taylor-Rodriguez D, Ealy AD, Ayres H, Lima FS, Martinez N, Thatcher WW, Santos JEP. Effects of progesterone concentrations and follicular wave during growth of the ovulatory follicle on conceptus and endometrial transcriptome in dairy cows. J Dairy Sci. 2022 105:S0022-0302(21)00928-0. doi: 10.3168/jds.2021-20193.   Byrd MKH, Arneson AG, Soffa DR, Stewart JW, Rhoads ML. Human continuous glucose monitors for measurement of glucose in dairy cows. J Dairy Sci Comm. 2022, 3(1): 78-83. https://doi.org/10.3168/jdsc.2021-0147.   Ealy AD, Speckhart SL, Wooldridge LK. Cytokines that serve as embryokines in cattle. Animals 2021, 11(8), 2313; <a href="https://doi.org/10.3390/ani11082313">https://doi.org/10.3390/ani11082313</a>   Fontes PLP, Oosthuizen N, Ciriaco FM, Sanford CD, Canal LB, Cooke RF, Pohler KG, Henry DD, Mercadante VRG, Ealy AD, Johnson SE, DiLorenzo N, Lamb GC. Effects of nutrient restriction on the metabolic profile of Bos indicus-influenced and Bos taurus suckled beef cows. Animal. 2021 Mar;15(3):100166. doi: 10.1016/j.animal.2020.100166.   Kott ML, Pancini S, Speckhart SL, Kimble LN, White RR, Stewart JL, Johnson SE, Ealy AD. Effects of mid-gestational L-citrulline supplementation to twin-bearing ewes on umbilical blood flow, placental development and lamb production traits. Transl Anim Sci. 2021 Jun 9;5(3):txab102. doi: 10.1093/tas/txab102. Owens CE, Huffard HG, Nin-Velez AI, Duncan J, Teets CL, Daniels KM, Ealy AD, James RE, Knowlton KF, Cockrum RR. Microbes of various maternal body systems are predictive of calf digestive bacterial ecology. Animals. 2021 Jul 26;11(8):2210. doi: 10.3390/ani11082210.   Seekford ZK, Wooldridge LK, Dias NW, Timlin CL, Sales AF, Speckhart SL, Pohler KG, Cockrum RR, Mercadante VGR, Ealy AD. Interleukin-6 supplementation improves post-transfer embryonic and fetal development of in vitro-produced bovine embryos. Theriogenology. 2021 Aug;170:15-22. doi: 10.1016/j.theriogenology.2021.04.004. Epub 2021 Apr 28.   Timlin CL, Lynn A, Wooldridge LK, Uh K, Ealy AD, White RR, Lee K, Mercadante VRG. Physical parameters of bovine activated oocytes and zygotes as predictors of development success. Zygote. 2021 Mar 19:1-7. doi: 10.1017/S0967199421000058.   Wooldridge LK, Ealy AD. Interleukin-6 promotes primitive endoderm development in bovine blastocysts. BMC Dev Biol. 2021 Jan 12;21(1):3. doi: 10.1186/s12861-020-00235-z.   Wooldridge LK, Ealy AD. Leukemia inhibitory factor stimulates primitive endoderm expansion in the bovine inner cell mass. Front Anim Sci 2021. 2:796489. doi: 10.3389/fanim.2021.796489   Przygrodzka E, Monaco CF, Plewes MR, Li G, Wood JR, Cupp AS, and Davis JS (2021) Protein kinase A and 5’ AMP-activated protein kinase signaling pathways exert opposite effects on induction of autophogy in luteal cells. Front Cell Dev Biol, 9:723563, PMID: 34820368    McFee RM, Romereim SM, Snider AP, Summers AF, Pohlmeier WE, Kurz SG, Cushman RA, Davis JS, Wood JR, and Cupp AS (2021). A high-androgen microenvironment inhibits granulosa cell proliferation and alters cell identity. Mol Cell Endocrinol, 531:111288, PMID: 33905753    Snider AP, Romereim SM, McFee R, Summers AF, Pohlmeier WE, Kurz SG, Davis JS, Wood JR, and Cupp AS (2021) Transcriptomic data of bovine ovarian granulosa cells of control and High A4 cows. Data in Brief, 37:107217, PMID: 34189206, PMCID: PMC8220326    Nafziger S, Tenley S, Summers AF, Abedal-Majed MA, Hart M, Bergman J, Kurz SG, Davis JS, Wood JR, Cupp AS (2021). Attainment and maintenance of pubertal cyclicity may predict reproductive longevity in beef heifers. Biol Reprod 104:1360-1372, PMID 33709137  Bidne KL, Rister AL, McCain AR, Hitt BD, Dodds ED, and Wood JR. (2021) Maternal obesity alters placental lysophosphatidylcholine, lipid storage, and the expression of genes associated with lipid metabolism. Biol Reprod 104: 197-210, PMID: 33048132, PMCID: PMC7946805 Editor’s choice, Cover Image   Rial C., Laplacette A.L., and Giordano J.O.* 2022. Effect of a targeted reproductive management program designed to prioritize insemination at detected estrus and optimize days to insemination on the reproductive performance of lactating dairy cows. J. Dairy Sci. (In Press)   Giordano J. O. *, E. M. Sitko, C. Rial, M. M. Perez and G. E. Granados. 2022. Use of multiple biological, management, and performance data for the design of targeted reproductive management strategies for dairy cows. J. Dairy Sci. 105:4669-4678. <a href="https://doi.org/10.3168/jds.2021-21476">https://doi.org/10.3168/jds.2021-21476</a>   Schilkowsky E. M., G. E. Granados, E M. Sitko, M. Masello, M. M. Perez, J. O. Giordano*. 2021. 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Does the phytoestrogen Daidzein stimulate angiogenic factors in bovine steroidogenic luteal cells? 56th Annual Meeting of the Society for the Study of Reproduction, Spokane, Washington.   da Silva M, Oli N, Gambonini F, Pate J, Ott T. 2021. Changes in immune cell populations and phenotype in peripheral blood leukocytes of dairy cows and heifers during early pregnancy. Society for the Study of Reproduction. Annual Meeting, St Louis, MO, Dec 12-15th Hughes CHK, Pate JL. 2022. MicroRNA may regulate luteal cell susceptibility to cell death during acquisition of luteolytic capacity. Int Cong Anim Reprod. Bologna, IT Peterson, K. D., M. A. Oliver, J. L. Edwards, R. R. Payton, T. M. Prado, L. G. Strickland and D. J. Mathew. 2022. Utilization of a conceptus-endometrial co-culture system to investigate endometrial gene expression and histotroph dependent on conceptus origin. 19th International Conference on Animal Reproduction (ICAR), Bologna, Italy.   Ault-Seay, T. B., K. J. Brandt, M. T. Henniger, D. J. Mathew, S. E. Moorey, F. N. Schrick, K. G. Pohler, J. D. Rhinehart, L. G. Schneider, K. J. McLean, P. R. Myer. 2021. Bacterial Communities in the Uterus and Rumen of Beef Heifers throughout Development with Protein Supplementation. American Society of Animal Science (ASAS) Southern Section. Fort Worth, TX, USA.   Payton, R.R., A.M. Saxton and J.L. Edwards. 2021. Differences in the responsiveness of maturing bovine oocytes to an acute heat stress depending on month of the year. 54th Annual Meeting of the Society for the Study of Reproduction (SSR), St Louis, MO.   Read, C.C., J.L. Edwards, F.N. Schrick, J.D. Rhinehart, R.R. Payton, S.R. Campagna, H.F. Castro, J.L. Klabnik, and S.E. Moorey. Influence of pre-ovulatory follicle size on the follicular fluid metabolome in lactating beef cows. 54th Annual Meeting of the Society for the Study of Reproduction (SSR), St Louis, MO. Horn, E.J., C.C. Read, J.L. Edwards, F.N. Schrick, J.D. Rhinehart, R.R. Payton, S.R. Campagna, H.F. Castro, J.L. Klabnik, and S.E. Moorey. Pre-ovulatory follicular fluid and serum metabolome profiles in beef cows with thin, moderate, and obese body condition. 54th Annual Meeting of the Society for the Study of Reproduction (SSR), St Louis, MO. Hessock, E.A., M.F. Smith, T.W. Geary, K.G. Pohler, J.A. Green, S.K. Behura, A.K. Stokes, and S.E. Moorey. Transcriptome profiles of in vivo matured bovine cumulus-oocyte complexes reveal unique gene expression between the oocyte and cumulus cells. 54th Annual Meeting of the Society for the Study of Reproduction (SSR), St Louis, MO. Oliver, M.A., K.D. Peterson, S. Bhandari, R.R. Payton, L.G. Schneider, J.L. Edwards, and D.J. Mathew. The effect of endometrial cell conditioned media on in vitro cultured bovine embryo development. 54th Annual Meeting of the Society for the Study of Reproduction (SSR), St Louis, MO. Mathew, D.J., H.L. Chaney, L.F. Grose, G. Charpigny, S.K. Behura, I.M. Sheldon, J.G. Cronin, T.E. Spencer, and P. Lonergan. Effect of galectin-1 on bovine endometrial epithelial and fibroblast cell transcriptomes in 3D cell culture. 54th Annual Meeting of the Society for the Study of Reproduction (SSR), St Louis, MO. Peterson, K.D., M.A. Oliver, S. Bhandari, S.E. Moorey, and D.J. Mathew. 2021. The effect of small and large ovarian follicle fluid on oviduct cell gene expression in cattle. American Dairy Science Association 2021 Annual meeting. Virtual.   Oliver, M.A., K.D. Peterson, S. Bhandari and D.J. Mathew. Effect of progesterone on bovine endometrial cell expression of select transporters and growth factors. American Dairy Science Association 2021 Annual meeting. Virtual.   Arneson AG, Stewart JW, Harrod MK, Newberne HM, Burgess MK, Jordan JA, White RR, Ealy AD, El-Kadi SW, Rhoads RP, Rhoads ML. Impact of heat stress and glycemic state on plasma γ- aminobutyric acid (GABA) in lactating Holstein cows. 2021. J Dairy Sci. 104(suppl 1): 173.   Oliver MA, Speckhart SL, Ealy AD. Human recombinant interleukin-6 promotes inner cell mass development in bovine embryos. Society for the Study or Reproduction Annual Meeting, Spokane WA.   Speckhart SL, Farrell K, Wooldridge LK, Biase FH, Lee K, Ealy AD. Interleukin-6 signal transducer is involved in blastocyst development in cattle. 2022 Society for the Study or Reproduction Annual Meeting, Spokane WA.   Stewart JW, Newberne HM, Harrod MK, Arneson AG, Negron-Perez VM, White RR, El-Kadi SW, Ealy AD, Rhoads RP, Rhoads ML. Heat stress and glycemic state alter milk production and composition in Holstein dairy cows. 2021. J Dairy Sci. 104(suppl 1): 173.   Cunha TO, Statz LR, Domingues RR, Andrade JPN, Wiltbank MC, Martins JPN. Accessory corpus luteum induced by human chorionic gonadotropin on day 7 or days 7 and 13 of the estrous cycle affected follicular and luteal dynamics and luteolysis in lactating Holstein cows. J Dairy Sci 2022; 105:2631-2650. Toledo MZ, Stangaferro ML, Gennari RS, Barletta RV, Perez MM, Wijma R, Sitko EM, Granados G, Masello M, Van Amburgh ME, Luchini D, Giordano JO, et al. Effects of feeding rumen-protected methionine pre- and postpartum in multiparous Holstein cows: Lactation performance and plasma amino acid concentrations. J Dairy Sci 2021; 104:7583-7603. Stangaferro ML, Toledo MZ, Gennari RS, Perez MM, Gamarra CA, Sitko EM, Monteiro PLJ, Jr., Masello M, Prata AB, Granados GE, Van Amburgh ME, Luchini D, et al. Effects of feeding rumen-protected methionine pre- and postpartum on reproductive outcomes of multiparous Holstein cows. J Dairy Sci 2021; 104:11210-11225. ** Monteiro PLJ, Sartori R, Canavessi AMO, Melo LF, Motta JCL, Consentini CEC, Wiltbank MC. Accessory corpus luteum regression during pregnancy I: timing, physiology, and P4 profiles. Reproduction 2021; 162:473-482. Monteiro PLJ, Gonzales B, Drum JN, Santos JEP, Wiltbank MC, Sartori R. Prevalence and risk factors related to anovular phenotypes in dairy cows. J Dairy Sci 2021; 104:2369-2383. Monteiro PLJ, Gamarra CA, Genari RS, Prata AB, Barletta RV, Duran PG, Canavessi AMO, Sartori R, Wiltbank MC. Accessory corpus luteum regression during pregnancy II: reproductive outcomes. Reproduction 2021; 162:483-495. Mezera MA, Li W, Wiltbank MC. Pregnancy-induced changes in the transcriptome of the bovine corpus luteum during and after embryonic interferon-tau secretiondagger. Biol Reprod 2021; 105:148-163. Mezera MA, Li W, Liu L, Meidan R, Penagaricano F, Wiltbank MC. Effect of natural pre-luteolytic prostaglandin F2alpha pulses on the bovine luteal transcriptome during spontaneous luteal regressiondagger. Biol Reprod 2021; 105:1016-1029. Cardoso Consentini CE, Wiltbank MC, Sartori R. Factors That Optimize Reproductive Efficiency in Dairy Herds with an Emphasis on Timed Artificial Insemination Programs. Animals (Basel) 2021; 11. Alves R, Silva MA, Consentini CEC, LO ES, Folchini NP, Oliva AL, Prata AB, Goncalves JRS, Wiltbank MC, Sartori R. Hormonal combinations aiming to improve reproductive outcomes of Bos indicus cows submitted to estradiol/progesterone-based timed AI protocols. Theriogenology 2021; 169:89-99. Mezera MA, Lauber MR, Beard AD, Cabrera EM, Wiltbank MC, Fricke PM. Effect of route of administration of dinoprost tromethamine on plasma profiles of 13,14-dihydro-15-keto-prostaglandin F2α and progesterone in lactating Holstein cows. JDS Communications 2021; 2:421-425. Toledo MZ, Nienow C, Luchini D, Apelo SIA, Wiltbank MC. Quantification of bovine plasma amino acids via liquid chromatography-electrospray ionization-mass spectrometry: Comparison of underivatized and precolumn derivatized methods. JDS Communications 2021; 2:227-232. Ermisch AF, Bidne KL, Bochantin KA, Wood JR (2021) Ovarian inflammation increases oocyte maternal mRNAs during maturation and alters expression of cumulus regulatory genes resulting in reduced developmental competence. 54th Annual Meeting of the Society for the Study of Reproduction, St. Louis, MO, Oral presentation    Bell B, Kurz SG, Abedal-Majed M, Springman S, Hart M, Snider AP, Wood JR, Davis JS, Cupp AS (2021) Anti-Mullerian Hormone and pro-inflammatory cytokines contribute to inflammation and follicle arrest while Vascular Endothelial Growth Factor A isoforms may rescue follicle progression in High A4 cow ovarian microenvironment. 54th Annual Meeting of the Society for the Study of Reproduction, St. Louis, MO, Oral presentation    Keane JA, Snider AP, Fudolig MA, Sutton CM, Bochantin KA, Bergman JW, Kurz SG, Hanford KJ, Wood JR, and Cupp AS (2021) Reductions in hematocrit and hemoglobin concentrations in non-cycling pubertal classification beef heifers may contribute to delayed pubertal attainment. 54th Annual Meeting of the Society for the Study of Reproduction, St. Louis, MO, Poster.   Rial C., A. L. Laplacette, and J. O. Giordano*. 2021. Effect of first-service management programs including AI at detected estrus based on automated estrus alerts and estrus occurrence during the voluntary waiting period on reproductive outcomes of dairy cows. J. Dairy Sci. Vol. 104 (E-Suppl. 1).   Giordano, J.O*. 2021. Use of multiple biological, management, and performance data for the design of targeted reproductive management strategies for dairy cows. J. Dairy Sci. 104 (E-Suppl. 1).   Sitko, E. M., Pérez, M. M., Granados, G. E., Masello, M., J.O. Giordano*. 2021. Effect of reproductive programs that prioritized timed AI or AI at detected estrus on cash flow of primiparous dairy cows. J. Dairy Sci.; 104. (E-Suppl. 1).   Theses/Dissertations   Crites, Benjamin Ryne, "THE EFFECTS OF FORM OF SELENIUM ON THE BOVINE CORPUS LUTEUM, UTERINE ENDOMETRIUM, AND DEVELOPMENT OF THE CONCEPTUS" (2021)”. Theses and Dissertations--Animal and Food Sciences. 135. https://uknowledge.uky.edu/animalsci_etds/135 https://doi.org/10.13023/etd.2021.424   Helmers, AC. 2022. The role of microRNA, complement proteins and immune cells in bovine luteal function and rescue. PhD Dissertation. The Pennsylvania State University. (Pate) Oli, N. 2022. Effects of the estrous cycle and early pregnancy on proportions and functions of peripheral immune cells in dairy cows and heifers. M.S. Thesis. The Pennsylvania State University. (Ott) Master’s Thesis: “Increasing vitamin D levels to improve fertilization rates in cattle.” Vanessa Peixoto de Souza, M.S., D.V.M. 06/14/2021. Major Professors C. Estill and C. Bishop. Casey Read. Effects of pre-ovulatory follicle physiological status on oocyte metabolic capacity. (PhD Dissertation, 2022; SEM PhD student). Mary Ali Oliver. The effect of endometrial cell conditioned media on in vitro cultured bovine embryo development. (MS Thesis, 2021; DJM MS student) Sarah Singleton. Novel postpartum reproductive phenotypes in beef cattle. (Undergraduate Chancellor’s Honor’s Thesis, 2021; SEM undergraduate student) Alicia Arneson (2021). Evaluating the impact of heat stress and altered glycemic state on plasma gamma-aminobutyric acid (GABA) in lactating Holstein cows (MS thesis).   Mary-Kathryn Harrod-Byrd (2021). Serial measurements of circulating glucose and luteinizing hormone concentrations in lactating dairy cattle (MS Thesis).   Michelle L. Kott (2021). Effects of mid-gestational L-citrulline supplementation to twin-bearing ewes on umbilical blood flow, placental development, and lamb production traits (MS Thesis).   Dallas Soffa (2022). Effects of feed additives on uterine morphology and selected reproductive attributes (MS thesis).   Jacob Stewart (2021). Physiological consequences of exposure to heat stress and the mycotoxin zearalenone (MS thesis).

Impact Statements

In addition to Hatch Multistate Funds, these studies were supported by: USDA NIFA/AFRI Oregon State University Animal and Rangeland Sciences Department Funds Oregon State University Agricultural Research Foundation Oregon Beef Council OARDC SEEDS Early career investigator – The Ohio State University CFAES Honors Will C. Hauk Endowment Grant – Undergraduate student - The Ohio State University National Institute of Environmental Health Science Iowa State University Nutritional Sciences Council Martin Fund National Pork Board Texas A&M University - Triads for Transformation American Simmental Association University of Tennessee AgResearch Instrumentation Grants program NIH-R21 Binational Agriculture Research and Development (BARD) Office of Research and Economic Development, Univ Nebraska-Lincoln Bill and Melinda Gates Foundation NIH/NIGMS-sponsored Ruth L. Kirschstein National Research Service Award (NRSA) Institutional Research Training (T32)