Brief Summary of Minutes of Annual Meeting

Generated information on micro RNAs and zinc finger proteins which expand our knowledge of regulation of oocyte maturation and early embryogenesis. Evaluated strategies that increase the proportion of cows inseminated in estrus (important to numerous farms) or via timed artificial insemination (TAI) with the finding that it is imperative that a TAI protocol is included immediately after completion of the period intended to inseminate cows in estrus. Identified the impact that thermal stress has on ovarian signaling, resulting in a mechanistic map being developed, upon which strategies to ameliorate seasonal infertility can be based. Germinal vesicle stage and mature bovine oocytes have been collected and an assay for quantification of sonic hedgehog messenger RNA designed. Discovered that the effects of estradiol on bovine follicle formation and capacity to activate are mediated through binding to a nuclear receptor, with evidence particularly strong for a role for ESR2. Activin A had effects on both early follicular development (stimulation of follicle formation and activation) and also on steroidogenesis by fetal bovine ovarian tissue, with effects on progesterone, but not estradiol, production. Determined that changes in microRNA expression in the corpus luteum may facilitate luteal rescue during maternal recognition of pregnancy (MRP). The miRNA that change during MRP target pathways involved in apoptosis and immune response. Discovered that acute zinc depletion promotes progesterone accumulation in media from cumulus cells or luteal tissue cultured in vitro. Found that during early pregnancy in cattle, increased CD47 expression induces inhibitory signals known to bring about cell-cell adhesion and T cell inactivation. Determined that maturation of bovine cumulus oocyte complexes in media containing follicular fluid improves cumulus cell expansion without affecting the outcome of in vitro fertilization Transcriptomal analysis examining the effect of stage of the cycle on epithelial cell function, specifically within the ampulla and isthmus of the bovine oviduct has been completed. Established novel animal models of metabolic stress which have been used to generate changes in knowledge regarding the steroids which are produced by somatic cells of the ovary and changes in oocyte gene expression associated with reduced oocyte quality. Blood samples and phenotypic data on reproductive performance have been collected from lactating dairy cows at 3 stations (NY, PA, and NH), DNA was extracted and cows genotyped for an AluI polymorphism in the growth hormone receptor (GHR) gene. Determined the physiological responses to cows that were treated with the Double-Ovsynch timed AI protocol. Determined that increased dose of PGF in the Double-Ovsynch protocol increased CL regression and fertility in multiparous dairy cows. Determined that increased dose of GnRH in Double-Ovsynch increased ovulation but did not increase fertility. Determined the effect of acute feed restriction on metabolic and reproductive hormones. Determined that acute feed restriction improved fertilization and embryo quality in cows that had were superovulated using an FSH product with higher LH content. However, using an FSH product with higher LH content compared to a low LH product was inhibitory to fertilization and embryo quality in late lactation cows that were full fed. A practical in vitro approach for the investigation of bovine granulosa cells from small follicles has been developed. Discovered that physiological concentration of estradiol and FGF2 may be regulators of CCN1 expression in a granulosa cell line.

Peer-reviewed research articles: Wang, L., Yao, J. 2014. The microRNAs important for ovarian and early embryonic development in cattle. Turkish Journal of Veterinary and Animal Sciences. 38: 599-605. Gupta, P.S.P., Folger, J.K., Rajput, S., Lv, L., Yao, J., Ireland, J., Smith, G.W. 2014. Regulation and regulatory role of WNT signaling in potentiating FSH action during bovine dominant follicle selection. PLoS ONE 9(6): e100201. Holler, T.L., Dean, M., Taylor, T., Poole, D.H., Thonney, M.L., Thomas, D.L., Pate, J.L., Whitley, N., Dailey, R.A., Inskeep, E.K. 2014. Effects of service sire on prenatal mortality and prolificacy in ewes. J. Anim. Sci. 92:3108-3115. Ganesan, S., Keating, A.F. 2014. Phosphoramide mustard exposure induces DNA adduct formation and the DNA damage repair response in rat ovarian granulosa cells. Toxicol Appl Pharmacol. 282(3):252-25. Ganesan, S., Nteeba, J., Keating, A.F. 2014. Enhanced susceptibility of ovaries from obese mice to 7,12-dimethylbenz[a]anthracene-induced DNA damage. Toxicol Appl Pharmacol. 281(2):203-210. Nteeba, J., Ganesan, S., Keating, A.F. 2014. Progressive obesity alters ovarian folliculogenesis with impacts on pro-inflammatory and steroidogenic signaling. Biol Reprod. 91(4):86. Madden, J.A., Keating, A.F. 2014. Ovarian xenobiotic biotransformation enzymes are altered during phosphoramide mustard-induced ovotoxicity. Toxicol Sci. 141(2):441-52. Baumgard, L.H., Keating, A.F., Ross, J.W., Rhoads, R.P. 2015. Effects of heat stress on the immune system, metabolism and nutrient partitioning: implications on reproductive success. Rev. Brazil Reprod. Anim. 39: 173-183. Maalouf, S.W., Liu, W.S., Albert, I., Pate, J.L. 2014. Regulating life or death: Potential role of microRNA in rescue of the corpus luteum. Mol Cell Endocrinol 398:78-88. Walusimbi, S.S., Pate, J.L. 2014. Luteal cells from functional and regressing corpora lutea differentially alter the function of gamma-delta T cells. Biol. Reprod. 90(6):140, 1-10. Ott, T.L., Kamat, M.M., Vasudevan, S., Townson, D.H., Pate, J.L. 2014. Maternal immune responses to conceptus signals during early pregnancy in ruminants. Anim. Reprod. 11(3): 237-245. Ott, T.L., Dechow, C., O’Connor, M.L. 2014. Advances in reproductive management: pregnancy diagnosis in ruminants. Anim. Reprod. 11(3): 207-216. Matthews, J.C., Zhang, Z., Patterson, J.D., Bridges, P.J., Stromberg, A.J., Boling, J.A. 2014. Hepatic transcriptome profiles differ among maturing beef heifers supplemented with inorganic, organic, or mixed (50% Inorganic:50% Organic) forms of dietary selenium. Biological Trace Element Research. 160:321-329. Matthews, J.C., Bridges, P.J. 2014. NutriPhysioGenomics applications to identify adaptations of cattle to consumption of ergot alkaloids and inorganic versus organic forms of selenium: altered nutritional, physiological and health states? Animal Production Science. 54:1594-1604. Pohlmeier, W.E., Xie, F., Kurz, S.G., Lu, N., Wood, J.R. 2014. Progressive obesity alters the steroidogenic response to ovulatory stimulation and increases the abundance of RNAs stored in the ovulated oocyte, Mol Reprod Dev 81(8): 735-747. Baier, S.R., Nguyen, C., Xie, F., Wood, J.R., Zempleni, J. 2014. MicroRNAs are absorbed in biologically meaningful amounts from nutritionally relevant doses of cow milk and affect gene expression in peripheral blood mononuclear cells, HEK-293 kidney cell cultures, and mouse livers J Nutr 144(10): 1495-1500. Summers, A.F., Pohlmeier, W.E., Brauer, V.M., Sargent, K.M., Kurz, S.G., McFee, R.M., Cushman, R.A., Cupp, A.S., Wood, J.R. 2014. Altered theca gene expression and ovarian follicular development in cows with follicular fluid androgen excess PLoS ONE 9(10) e110683. Vieira-Neto, A., Gilbert, R.O., Butler, W.R., Santos, J.E.P., Ribeiro, E.S., Vercouteren, M.M., Bruno, R.G., Bittar, J.H.J., Galvao, K.N.. Individual and combined effects of anovulation and cytological endometritis on the reproductive performance of dairy cows. J Dairy Sci. 97:5415-5425, 2014. Bender, R.W., Hackbart, K.S., Dresch, A.R., Carvalho, P.D., Vieira, L.M., Crump, P.M., Guenther, J.N., Fricke, P.M., Shaver, R.D., Combs, D.K., Wiltbank, M.C. 2014. Effects of acute feed restriction combined with targeted use of increasing luteinizing hormone content of follicle-stimulating hormone preparations on ovarian superstimulation, fertilization, and embryo quality in lactating dairy cows. Journal of Dairy Science 97:764-778. Ferraretto, L.F., Gencoglu, H., Hackbart, K.S., Nascimento, A.B., Dalla Costa, F., Bender, R.W., Guenther, J.N., Shaver, R.D., Wiltbank, M.C.. 2014. Effect of feed restriction on reproductive and metabolic hormones in dairy cows. Journal of Dairy Science 97:754-763. Nascimento, A.B., Souza, A.H., Keskin, A., Sartori, R., Wiltbank, M.C. 2014. Lack of complete regression of the Day 5 corpus luteum after one or two doses of PGF2? in nonlactating Holstein cows. Theriogenology 81:389-395. Pereira, M.H.C., Rodriguez, A.D.P., De Carvalho, R.J., Wiltbank, M.C., Vasconcelos, J.L.M. 2014. Increasing length of an estradiol and progesterone timed artificial insemination protocol decreases pregnancy losses in lactating dairy cows. Journal of Dairy Science 97:1-11. Wiltbank, M.C., Pursley, J.R. 2014. The cow as an induced ovulator: Timed AI after synchronization of ovulation. Theriogenology 81:170-185. Carvalho, P.D., Souza, A.H., Amundson, M.C., Hackbart, K.S., Fuenzalida, M.J., Herlihy, M.M., Ayres, H., Dresch, A.R., Vieira, L.M., Guenther, J.G., Grummer, R.R., Fricke, P.M., Shaver, R.D., Wiltbank, M.C. 2014. Relationships between fertility and postpartum changes in body condition and body weight in lactating dairy cows. Journal of Dairy Science 97:1-18. Wiltbank, M.C., Souza, A.H., Carvahlo, P.A., Cunha, A.P., Giordano, J.O., Fricke, P.M., Baez, G.M., Diskin, M.G. 2014. Physiological and practical effects of progesterone on reproduction in dairy cattle. Animal 8:70-81. Wiltbank, M.C., Garcia-Guerra, A., Carvalho, P.D., Hackbart, K.S., Bender, R.W., Souza, A.H., Toledo, M.Z., Baez, G.M., Surjus, R., Sartori, R. 2014. Effects of energy and protein nutrition in the dam on embryonic development. Animal Reproduction 11:168-182. Carvalho, P.D., Guenther, J.N., Fuenzalida, M.J., Amundson, M.C., Wiltbank, M.C., Fricke, P.M. 2014. Presynchronization using a modified Ovsynch protocol or a single gonadotropin-releasing hormone injection 7 d before an Ovsynch-56 protocol for submission of lactating dairy cows to first timed artificial insemination. Journal of Dairy Science 97:6305-6315. Book Chapters and Review Articles: Johnson, J.S., Abuajamieh, M., Sanz-Fernandez, M.V., Seibert, J.T., Stoakes, S.K., Nteeba, J., Keating, A.F., Ross, J.W., Rhoads, R.P., Baumgard, L.H. 2014. Thermal stress alters postabsorptive metabolism during pre- and postnatal development. Climate Change Impact on Livestock: Adaptation and Mitigation. Seijan, V. (ed.). Chapter 5, 61-79. Hoyer, P.B., Keating, A.F. 2014. Xenobiotic effects in the ovary: temporary versus permanent infertility. Drug Metabolism and Toxicology (Expert Opinion). 10(4):511-23. Conference papers and Abstracts: Rhoads, M.L. 2014. Implementing strategies to reduce heat stress in dairy herds. Proc. Reprod Action Cattle Symp. 4-8. Mouttham, L.L., Fortune, J.E., Comizzoli, P. 2014. Short- and long-term effects of cryoprotectant exposure and vitrification on follicular health and primordial follicle activation in vitro. Biology of Reproduction, Supplement, Proc. of the 47th annual meeting of the Soc. for the Study of Reproduction. Yang, MY, Cheong, S.H., Fortune, J.E. 2014. Bisphenol A inhibits estradiol production and alters abundance of mRNA for key steroidogenic enzymes and estrogen receptor in granulosa cells from bovine preovulatory follicles. Biology of Reproduction, Supplement, Proc. of the 47th annual meeting of the Soc. for the Study of Reproduction. Allen, J.J., Fortune, J.E. 2014. Formation and activation of follicles in fetal bovine ovarian cortex in vitro are inhibited by increasing endogenous estradiol and progesterone. Biology of Reproduction, Supplement, Proc. of the 47th annual meeting of the Soc. for the Study of Reproduction. Giordano, J.O. Estrus Detection Programs and Economics of Monitoring Systems. Dairy Cattle Reproduction Council Annual Meeting. Salt Lake city, Utah. November 12-13th, 2014. Wijma R., Stangaferro, M.L., Branen, J.R., Sasser, R.G., Giordano, J.O. 2014. Altered Ovarian Dynamics in Lactating Dairy Cows Undergoing Embryonic Mortality. Abstract in Press. J. Dairy Sci. Volume 97, E-Supplement 1. Giordano J.O., Watters, R.D., Wijma, R., Stangaferro, M.L. 2014. Reproductive Performance of Lactating Dairy Cows after Resynchronization with Ovsynch or a Program Aimed to Maximize Artificial Insemination in Estrus and Fertility of Timed Artificial Inseminations based on Ovarian Structures. J. Dairy Sci. Volume 97, E-Supplement 1. Giordano, J.O. Reproductive Management: Practical Strategies and Research Update. Northern New York Dairy Institute. Watertown, Lowville, and Chazy, New York. December 16th, 17th, and 18th, 2014. Giordano, J.O. Ohio Dairy Health and Management Certificate Program: Dairy Cattle Reproduction with Emphasis on Transition Cow Management in Confinement Systems. 1. Activity Monitors: Managing 1st and 2nd and Subsequent AI Services. College of Veterinary Medicine, The Ohio State University, Columbus, Ohio. August 7th, 2014. Giordano, J.O. Reproductive Physiology Update. Summer Dairy Institute. College of Veterinary Medicine, Cornell University. Ithaca, New York. June 24th, 2014. Giordano, J.O. Dairy Health Management Certificate Program. 1. Reproductive Management I: Practical Strategies and Research Update, 2. Pregnancy Losses and Non-Pregnancy Diagnosis, 3. Reproductive Management II: Integrating Activity Monitoring into Reproductive Management Programs. Ontario College of Veterinary Medicine, Guelph Ontario. June 3rd to June 5th, 2014. Giordano, J.O. Dairy Day at Miner Institute. Evaluating Reproductive Management Strategies and their Profitability. Chazy, NY. December 3rd, 2013. Keating, A.F. 2014. Impact of insulin on ovarian function. J. Anim. Sci. Vol 92. E-Suppl. 2: 46. Sholar, J.F., Colpoys, J.D., Gabler, N.K., Keating, A.F., Millman, S.T., Siegford, J.M., Johnson, A.K. 2014. Gilt approachability to a human when selected for feed efficiency. J. Anim. Sci. Vol 92. E-Suppl. 2: 114. Colpoys, J.D., Abell, C.E., Gabler, N.K., Keating, A.F., Millman, S.T., Siegford, J.M., Johnson, A.K. 2014. Gilt behavioral responsiveness to a novel object when selected for feed efficiency. Johnson, J.S., Abuajamieh, M., Sanz Fernandez, M.V., Seibert, J.T., Stoakes, S.K., Keating, A.F., Ross, J.W., Selsby, J.T., Rhoads, R.P., Baumgard, L.H. 2014. The impact of in utero heat stress and nutrient restriction on progeny body composition. J. Anim. Sci. Vol 92. E-Suppl. 2: 583. Baumgard, L.H., Ross, J.W., Gabler, N.K., Lonergan, S.M., Keating, A.F., Selsby, J.T., Rhoads, R.P. 2014. Metabolic and health consequences of heat stress: Knowledge gaps and opportunities. J. Anim. Sci. Vol 92. E-Suppl. 2: 31. Nteeba, J., Ganesan, S., Keating, A.F. 2014. Progressive obesity alters the expression of ovarian NF-?B and steroidogenic pathway members in mice. Proceedings of the 47th Annual Meeting of the Society for the Study of Reproduction, Grand Rapids, MI. Madden, J.A., Keating, A.F. 2014. Phosphoramide mustard-induced autophagy in the neonatal rat ovary. Proceedings of the 47th Annual Meeting of the Society for the Study of Reproduction, Grand Rapids, MI. Ganesan, S., Nteeba, J., Keating, A.F. 2014. Impact of 7,12-dimethylbenz[a]anthracene exposure and obesity on ovarian connexin gap junction proteins in mice. Proceedings of the 47th Annual Meeting of the Society for the Study of Reproduction, Grand Rapids, MI. Madden, J.A., Keating, A.F. 2014. Phosphoramide mustard-induced changes in glucose metabolism in the neonatal rat ovary. Society of Toxicology annual meeting. Ganesan, S., Keating, A.F. 2014. Phosphoramide mustard-induced DNA adduct formation and DNA damage repair response induction in rat ovarian granulosa cells. Society of Toxicology annual meeting. Nteeba, J., Keating, A.F. 2014. Temporal impact of obesity on expression of genes encoding enzymes involved in ovarian xenobiotic biotransformation. Society of Toxicology annual meeting. Maalouf, S., Liu, W-S., Albert, I., Pate, J. 2014. MicroRNA of the corpus luteum during maternal recognition of pregnancy. Society for the Study of Reproduction. Abstract 552. Steinberger L, Pate, J.L. 2014. Isolation and characterization of macrophages in the bovine corpus luteum. Amer J Reprod Immunol 71 (Suppl. 1):81 Branham, K., Waugh, S., Pate, J.L. 2014. In vitro differentiation of bovine macrophages. Amer J Reprod Immunol 71 (Suppl. 1):37. Oh, J., Walusimbi, S., Giallongo, F., Weeks, H., Frederick, T., Hristov, A.N., Pate, J.L., Elias, R.J., Tao, L., Wall, E. H. 2014. Effect of dietary supplementation of Capsicum extract on immune responses, blood cell counts, blood chemistry, and oxidative stress markers in lactating dairy cows. J. Dairy Sci. 97(E-Suppl. 1):103. Townson, D.H., Diaz, F.J., Ocon-Grove, O., Johnson, A.L. 2014. A practical in vitro approach for the investigation of bovine granulosa cells from small follicles. Proceedings of the 47th Annual Meeting of the Society for the Study of Reproduction, Grand Rapids, MI. Sun, T. Diaz, F.J. 2014. The Effect of Oocytes and EGF on Hippo Signaling Pathway in Cumulus Cells During Ovulatory Transition in Mice. Proceedings of the 47th Annual Meeting of the Society for the Study of Reproduction, Grand Rapids, MI. Zhao, Y., Ning, G., Diaz, F.J., Ocon-Grove, O.M., Liu, W.S. 2014. PRAMEY Is a Novel Male Germ Cell-Specific Protein Involved in Acrosome Biogenesis and the Block to Polyspermy during Fertilization in Cattle. Proceedings of the 47th Annual Meeting of the Society for the Study of Reproduction, Grand Rapids, MI. McCracken, V.L.*, Wood, A.Y., Zezeski, A.L., Wiegert, J.G., Lugar, D.W., Scheffler, J.M., El-Kadi, S.W., Ealy, A.D., Rhoads, M.L. 2014. Porcine pre-pubertal reproductive tract development is altered by consumption of high fructose but not high fat diets. Biol. Reprod. Matthews, J.C., Bridges, P.J. 2014. NutriPhysioGenomics: What do they tell us about adaptation of cattle to nutritional, physiological and health challenges? Proceedings of the 2014 Joint International Symposium on the Nutrition of Herbivores/International Symposium on Ruminant Physiology. Coticchia, C., Miseirvitch, J., Cummings, C., Davis, J.S., Tsang, P.C.W., Moses, M.A. (2014). Regulation of cysteine rich 61-connective tissue growth factor-nephroblastoma overexpressed (CCN1) in granulosa cells. 47th Meeting of the Society for the Study of Reproduction in Grand Rapids, MI. Fernandes, N., Jaskiewicz, N.M., Chu, F., Townson, D.H. (2014) Determining the functional role of keratin filament in apoptosis via the PI3K/Akt signaling pathway using LTQ Orbitrap MS/MS analysis. Journal for the American Society for Mass Spectrometry Annual Conference, 25(1): 142., Boston, MA. Xie, F., Timme, K., Rifer, J.L., Wood, J.R. (2014) Diet-induced obesity increases of Pou5f1 and Dppa3 in growing and mature oocytes are due to increased STAT3 signaling. 11th Annual Gilbert Greenwald Symposium on Reproduction, Kansas City, KS. Romereim, S., Summers, A.F., Pohlmeier, W.E., McFee, R., Spuri-Gomes, R., Kurz, S.G., Cushman, R.A., Davis, J.S., Wood, J.R., Cupp, A.S. (2014) Differential regulation of cell cycle and immune response networks in bovine granulosa cells with excess intrafollicular androstenedione. 11th Annual Gilbert Greenwald Symposium on Reproduction, Kansas City, KS. Carvalho, P.D., Lobos, N.E., Toledo, M.Z., Trevisol, E., Santos, V.G., Barletta, R.V., Baez, G.M., Garcia-Guerra, A., Guenther, J.N., Souza, A.H., Luchini, D., Fricke, P.M., Shaver, R.D., Wiltbank, M.C. 2014. Changes in plasma methionine concentrations after administration of two different doses of rumen protected methionine. Abstract # 1539. Lobos, N. E., Broderick, G. A., Carvalho, P. D., Luchini, D. N., Shaver, R. D., Souza, A. H., Wiltbank, M. C. 2014. Amino acid analysis in dairy cow plasma by chloroformate derivatization and gas chromatography. Abstract # 1546. Carvalho, P.D., Sousa, A.H., Amundson, M.C., Hackbart, K.S., Dresch, A.R., Vieira, L.M., Guenther, J.N., Grummer, R.R., Shaver, R.D., Fricke, P.M., Wiltbank, M.C. 2014. Relationship between fertility and postpartum changes in body condition and body weight in lactating dairy cows. Abstract # 337. Souza, A.H., Carvalho, P.D., Drake, C.M., Shaver, R.D.,Wiltbank, M.C. 2014. Ration composition in Wisconsin dairy herds: Factors affecting fertility. Abstract # 1851. ADSAP. Carvalho, D., Fuenzalida, M. J., Ricci, A., Luchterhand, M., Mulcahy, J.M., Barletta, R.V., Baez, G.M., Santos, V.G., Amundson, M.C., Guenther, J.N., Sousa, A.H., Wiltbank, M.C., Fricke, P.M. 2014. Modifications to Ovsynch improve fertility during resynchronization: Evaluation of presynchronization with GnRH 6 days before Ovsynch and addition of a second PGF treatment. Abstract # 524. Wiltbank, M.C., Hansen, P.J., Santos, J.E.P. 2014. How knowledge of bovine reproductive physiology and development of reproductive technologies will help to feed a growing world population. Abstract # 72. Baez, G.M., Barletta, R.V., Ricci, A., Trevisol, E., Garcia-Guerra, A., Carvalho, P.D., Guenther, J.N., Cardoso, B.O., Toledo, M.Z., Wiltbank, M.C. 2014. Treatment with GnRH 5d after Breeding Preferentially Produces an Accessory CL Contralateral to the Pregnancy and this CL Generally Regresses Prior to Day 75 of Pregnancy in Lactating Dairy Cows. Abstract # 206. Baez, G.M., Barletta, R.V., Cardoso, B.O., Trevisol, E., Wiltbank, M.C. 2014. Uterine size matters for fertility in lactating dairy cows. Abstract # A079. Selected as Second-Place Abstract in Practical Reproduction. Souza, A.H. ,Narciso, C.D., Batista, E.O.S., Carvalho, P.D., Wiltbank, M.C. 2014. Effect of uterine environment on embryo production and fertility in cows. Abstract # 159. Wiltbank, M.C., Garcia-Guerra, A., Carvalho, P.D., Hackbart, K.S., Bender, R.W., Souza, A.H., Toledo, M.Z., Baez, G.M., Surjus, R.S., Sartori, R. 2014. Effects of energy and protein nutrition in the dam on embryonic development. Abstract # 168. Wiltbank, M.C., Baez, G.M., Vasconcelos, J.L.M., Pereira, M., Souza, A.H., Sartori, R., Pursley, J.R. 2014. The physiology and impact on fertility of the period of proestrus in lactating dairy cows. Abstract # 225. Theses/Dissertations: Bowdridge, E.C. 2014. The role of adenosine monophosphate activated kinase in luteal progesterone production. Doctoral dissertation, West Virginia University. 95 pages. Graham, M.R. 2014. Effects of lipopolysaccharide induced inflammatory response on early embryo survival in ewes. M.S. Thesis, West Virginia University. 84 pages. Akers, K.A. 2014. Changes in body condition from prepartum to postpartum on health and reproduction in dairy cattle. M.S. Thesis, West Virginia University In addition to Hatch Multistate Funds, these studies were supported by: Global Food Security Consortium and the Iowa Pork Producers Association to AFK. United States Department of Agriculture (NRICGP, #2008-35203-05989) to JEF. National Institute of Health research grant HD074831 to FJD. Agriculture and Food Research Initiative Competitive Grant no. 2012-67015-30212 from the USDA National Institute of Food and Agriculture to JLP. C. Lee Rumberger and Family Chair in Agricultural Science funds to JLP. National Pork Board, Virginia Horse Industry Board and John Lee Pratt Animal Nutrition Program to MLR.