NE2227: Contribution of Ovarian Function, Uterine Receptivity, and Embryo Quality to Pregnancy Success in Ruminants

(Multistate Research Project)

Status: Active

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SAES-422 Reports

Annual/Termination Reports:

[05/23/2023] [09/30/2024] [06/05/2025]

Date of Annual Report: 05/23/2023

Report Information

Annual Meeting Dates: 05/16/2023 - 05/17/2023
Period the Report Covers: 10/01/2022 - 09/30/2023

Participants

Dave Townson-URhode Island
Fransisco Diaz- Penn State
Cecily Bishop- Oregon State
Milo Wiltbank- UWisconsin
Yi Ren - Cornell
Amanda Patterson- UMissouri
Sarah Moorey-UTennessee
Dan Mathew-UTennessee
Soon Hun Cheong- Cornell
Jen Wood- UNebraska
Phil Bridges- UKentucky
Ron Butler- Cornell
Joy Pate- Penn State
Camilla Hughes- Penn State
Troy Ott- Penn State
Alvaro Garcia Guerra -Ohio State
Aileen Keating – Iowa State
Abby Maucieri – URhode Island
Kamilah Grant -USDA
Mark Mirando - USDA
Joanne Fortune -Cornell

Brief Summary of Minutes

Please see attached document

Minutes Attachment:

NE2227_Minutes_2023_PSU.docx

Accomplishments

Objective 1: Identify Mechanisms that Regulate Ovarian Function and Oocyte Quality during the Estrous Cycle   Identification of 22 metabolites that had positive relationship with serum estradiol concentration at the time of GnRH injection to induce ovulation. Determined a positive relationship between serum estradiol concentration at the time of GnRH injection to induce ovulation and oocyte ATP level ~18 hours later.   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.   Discovered that O-GlcNAcylation is expressed in KGN cells, an immortal granulosa cell tumor line that shares functional features of granulosa cells of mature, preovulatory follicles (e.g., FSH-responsiveness and estradiol production).   Demonstrated that manipulation of O-GlcNAcylation in KGN cells impairs cell proliferation and acutely perturbs oxidative phosphorylation, an effect observed previously with granulosa cells of bovine large antral follicles.   Preliminary experiments indicate inhibition of O-GlcNAc enhances estradiol production by primary cultures of bovine granulosa cells.   Determined the impact of altered insulin and glucose observed in dairy cattle experiencing heat stress (impact on the reproductive tract and elsewhere throughout the body. Recent work suggested that hypoglycemia during heat stress may be an important coping mechanism rather than a maladaptation to hyperthermia. Studies investigating the effect of hypoglycemia on follicle developmental dynamics and uterine function are currently underway.   Objective 2: Determine Factors Associated with Fertilization, Embryo Development, and Conceptus-Endometrial Interactions that Dictate Pregnancy Success   Using a day 16 conceptus-endometrial co-culture system, we are conducting studies to identify endometrial proteins and metabolites induced by IVP compared to IVD bovine embryos. Efforts are also underway to characterize the day 16 IVP and IVD bovine embryo transcriptome and secreted proteins/metabolites to better understand which conceptus secretory factors may be responsible for the altered endometrial transcriptome.    The gene codes for a KRAB-ZFP specifically expressed in bovine oocytes and early embryos and gene silencing experiments have demonstrated that ZNFO is required for early embryonic development in cattle. These results provide an essential step towards the identification of ZNFO regulated genes that play important roles during early embryonic development.   A study was completed to characterize three novel oocyte-specific lncRNAs, OOSNCR1, OOSNCR2 and OOSNCR3 using expression profiles generated from tissues, cells and embryos collected across the different stages of early embryonic development. Tissue expression data revealed detectable expression in only the fetal ovary for all three lncRNAs. Cell expression data revealed highest expression in oocytes with no significant difference seen between GV and MII oocytes. LncRNA expression was detectable but at lower levels in cumulus cells (CC) and even lower in theca (TC) and granulosa cells (GC). Expression data from the embryo panel suggests all three lncRNAs are maternally derived. Together, these data indicate a dynamic relationship among OOSNCR1, OOSNCR2 and OOSNCR3 and early reproductive events. Future studies aim to elucidate the functional roles of these lncRNAs during oocyte maturation and early embryonic development.   The role of ASIP in the bovine oocyte and early embryo was unknown. ATRN, MC3R, and MC4R were found to be expressed in both cumulus cells and oocytes. ATRN was also expressed in granulosa and theca cells and there was a significant effect of cell type as cumulus cells collected from MII COCs expressed ATRN at higher levels than granulosa and theca cells. Oocyte maturity level did not affect ATRN, MC3R, and MC4R expression levels.  There was a significant effect of embryo stage on ASIP expression. While ASIP was detected in each stage analyzed, ASIP levels were slightly reduced following oocyte maturation and remained at constant low levels until following completion of the embryonic genome activation at the 16-cell stage where it was observed ASIP levels increased in morula and blastocyst stage embryos.    Efforts are currently focused on exploring how the interleukin-6 (IL6) cytokine family acts as regulators of oocyte maturation, early embryo development, blastocyst formation, and early conceptus development in cattle. The work over the past year has provided strong evidence for implicating IL6 as a critical regulator of blastocyst development and inner cell mass formation. IL6 supplementation also improves conceptus elongation.   Exploration into how to modify our current bovine culture media formulations to improve embryo development and quality. Minor changes in formulation of SOF medium and the inclusion of an antioxidant cocktail are showing promise for improving the number and quality of transferable embryos.   Investigating the relationship between hypoglycemia during heat stress and pregnancy success on-farm.   Studies have determined how vitamin A and D supplementation in mid- to late gestation affects calving ease and birth weights of calves at parturition, as well as cow fertility at the following breeding. There was no effect of vitamin A and D treatment on calving ease or on birth weight, although calves had significantly elevated serum vitamin D in the treated group at time of birth, compared to controls, however these levels dropped to near control calves within one month of age.   Induction of pregnancy loss on day 35 of gestation in cattle results in luteolysis by gestation day ~45. During the second month of gestation basal prostaglandin F2α (PGF2α) is greater whereas pulse frequency and amplitude are similar to that at the expected time of luteolysis during the estrous cycle. It, however, is unclear whether PGF2α is responsible for luteolysis after induced conceptus demise. The aim, therefore, was to test the hypothesis that inhibition of PGF2α secretion (using flunixin meglumine) will delay luteolysis after induced conceptus demise. Luteolysis was identified in all animals, however, luteolysis tended to occur later in cows receiving flunixin meglumine (FM) than the untreated controls. In conclusion, inhibition of PGF2α secretion did not prevent but rather delayed luteolysis after induced conceptus demise, thus, providing partial support for the hypothesis that PGF2α is responsible for initiation of luteolysis after conceptus demise.   Objective 3: Develop and Evaluate Novel Reproductive Management Strategies and Technologies to Improve Reproductive Performance of Ruminants   Growth and nutrition during early development are major contributors to fertility and lifetime reproductive performance in cattle. Recent research findings demonstrate that changes in nutrition during gestation and early postnatal life result in functional and structural modifications in brain areas involved in the control of pubertal maturation in females. While these early modifications are likely to persist throughout life, their long-term effects remain unknown. Through a series of whole-animal, physiological, and molecular studies we have demonstrated that nutritional extremes during late gestation have only negligible effects on reproductive function in the heifer offspring. More specifically, maternal obesity or severe feed restriction during the last two trimesters of gestation did not alter age at puberty, pre-ovulatory follicle size, dominant follicle growth rate, CL size, endometrial thickness, antral follicle count, estrus expression, progesterone and estradiol concentrations during the estrous cycles, nor gonadotropin secretion in response to exogenous estradiol. These studies indicate that the adult reproductive phenotype in Bos indicus-influenced heifers is resilient to significant degrees of nutritional stress imposed during pre- and early postnatal periods, particularly if early gestation (first trimester) is avoided. These studies strongly suggest that adequate postnatal nutrition can overcome deficits programmed during late gestation in sexually-mature beef heifers. This information can guide management strategies for selection of replacement beef heifers.   A ruminant model of ovarian cryopreservation was investigated as part of an NIH-SIBR funded project in collaboration with 21st Century Medicine. The major goal of the study was to determine if a novel cryoprotectant along with a novel infusion and vitrification protocol would preserve most small follicle types on the ruminant ovary. This work found that perfusion techniques to vitrify whole ovaries are highly successful in preserving follicular function in ewes. Further studies are necessary to determine if these follicles produce fertilizable oocytes.   Presynchronization treatments administered before initiation of a CO-Synch type regimen are designed to improve ovulatory response and synchronization of follicular development. We conducted a study to determine the effect of various presynchronization strategies before initiation of a 6-day CO-Synch, on estrous expression and fertility to timed artificial insemination (TAI). Pregnancies per AI (P/AI) on D35 were greater for P4+PGF Presynch cows (66.7%) than for PGF Presynch (55.3%) and Control cows (50.9%). There were no differences in pregnancy loss between groups (5.9%). In conclusion, presynchronization with P4 and PGF prior to initiation of a 6-day CO-Synch treatment regimen improves fertility as a result of TAI in suckled beef cows.   Administration of FSH prior to ovum pick-up (OPU) in cattle improves in vitro embryo production (IVEP). Even though FSH is typically administered following a decreasing dose schedule, other dose schedules have not been evaluated. Thus, we evaluated the effect of FSH treatment pattern on ovarian response and IVEP. Pregnant Holstein heifers were randomly assigned to be administered p-FSH in a decreasing, constant, or increasing dose schedule, in a crossover design with a 14-day washout between sessions. Total number of follicles did not differ between decreasing, constant, and increasing groups. No differences were observed for number of viable COCs, number of COCs grade I, nor COC recovery rate. Cleaved oocytes, cleavage rate, and blastocyst rate did not differ between groups, resulting in a similar number of blastocysts per heifer. In conclusion, the pattern of administration for p-FSH prior to IVEP does not affect ovarian response, oocyte developmental competence nor embryo production.   Elucidated the underlying biology of differences in reproductive physiology of dairy cows with different genetic merit for fertility.   Generated evidence to support that cows of different genomic merit for fertility present different reproductive performance under common types of reproductive management.   Evaluated the response of dairy cows to different types of reproductive management strategies.   Identified associations between automated sensor monitoring systems data and the reproductive potential of dairy cattle.   Explored the value of combining genomic data with sensor and herd management data for predicting the future reproductive potential of cows.   Developed and tested reproductive management strategies that increase reproductive performance and improve herd management through use automated detection of estrus.   Described economic differences of cows of different genetic merit for fertility under different types of reproductive management programs.   Determined the economic value of days to pregnancy after calving (i.e., days open) on the profitability and survivability of dairy cows.   Developed data-analytic methods based on machine learning algorithms to predict the reproductive potential of dairy cows based on data collected by automated monitoring technologies and herd management and performance data.

Publications

Ault-Seay TB, Brandt KJ, Henniger MT, Payton RR, Mathew DJ, Moorey SE, Schrick FN, Pohler KG, Smith TPL, Rhinehart JD, Schneider LG, McLean KJ, Myer PR. Bacterial communities of the uterus and rumen during heifer development with protein supplementation. Front Anim Sci 2022. doi: 10.3389/fanim.2022.903909.   Ault-Seay TB, Moorey SE, Mathew DJ, Schrick FN, Pohler KG, McLean KJ, Myer PR. Importance of the female reproductive tract microbiome and its relationship with the uterine environment for health and productivity in cattle: A review. Front Anim Sci 2022.  doi: 10.3389/fanim.2023.1111636.   Ault-Seay TB, Payton RR, Moorey SE, Pohler KG, Schrick FN, Shepherd EA, Voy BH, Lamour KH, Mathew DJ, Myer PR, McLean KJ. Endometrial gene expression in response to lipopolysaccharide between estrous cycle phases and uterine horns in cattle. Front Anim Sci 2022. doi: 10.3389/fanim.2022.939876.   Bishop C, Selvaraj V, Townson DH, Pate JL, Wiltbank MC. History, insights, and future perspectives on studies into luteal function in cattle. J Anim Sci 2022  100: 1-15. doi.org/10.1093/jas/skac143   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 Jan;105(1):889-903. 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: 78-83.   Carr SN, Crites B, Pate JL, Hughes CHK, Matthews JC, Bridges PJ.  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. 2022. 12:313.    Chaney HL, Grose LF, LaBarbara JM, Sirk AW, Blancke AM, Sanchez JM, Passaro C, Lonergan P, Mathew DJ. Galectin-1 induces gene and protein expression related to maternal-conceptus immune tolerance in bovine endometrium. Biol Reprod. 2022. doi: 10.1093/biolre/ioab215.   Crites BR, Carr SN, Matthews JC, Bridges PJ. Form of dietary selenium affects mRNA encoding cholesterol biosynthesis and immune response elements in the early luteal phase bovine corpus luteum.  J Anim Sci. 2022. 100(7): doi:10.1093/jas/skac135.   Crites BR, Carr SN, Anderson LH, Matthews JC , Bridges PJ. 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. J Anim Sci.  2022. 100(7): doi:10.1093/jas/skac137.   Ealy AD, Pate JL, Ron Butler W. A synopsis of the NE1727 multistate project collection in the Journal of Animal Science. J Anim Sci. 2022 Jul 1;100(7):skac173. doi: 10.1093/jas/skac173.     García-Guerra A, Motta JCL, Hayden CB, Sala RV, Pereira DC, V. Absalon-Medina A, Moreno JF, Ross PJ. Advances in donor synchronization and superstimulation for OPU-IVEP:  the quest for enhanced oocyte quantity and quality. Proceedings of the Joint Convention of the American Embryo Transfer Association and the Canadian Embryo Transfer Association 2022, Pages 1-8.   Garza V, West SM, Cardoso RC. Review: Gestational and postnatal nutritional regulation of puberty and subsequent reproductive performance in heifers. Animal (accepted), 2022.   Hayden CB, Sala RV, Absalon-Medina VA, Motta JCL, Pereira D, Moreno JF, García-Guerra A. Synchronization of follicle wave emergence before ovarian superstimulation with FSH and ovum pick-up improves in vitro embryo production in pregnant heifers. Theriogenology 2022 188: 71-78.   Hardin, KN, dos Reis BR, Dias NW, Fiske DA, Mercadante VRG, Rhoads ML, Wilson TB, White RR. Growth and reproductive responses of heifers consuming endophyte-infected tall fescue seed with or without sodium bicarbonate supplementation. Appl Anim Sci. 2022 38(4):317-325. doi.org/10.15232/aas.2022-02273.   Horn, EJ, Read CC, Edwards JL, Schrick FN, Rhinehart JD, Payton RR, Campagna SR, Klabnik JL, Clark HM, Myer PR, McLean KJ, Moorey SE. Preovulatory follicular fluid and serum metabolome profiles in lactating beef cows with thin, moderate, and obese body condition. J Anim Sci. 2022. 1 (7): skac152. doi: 10.1093/jas/skac152.   Klabnik JL, Christenson LK, Pohler KG, Moorey SE, Rispoli LA, Gunewardena SSA, Payton RR, Schrick FN, Edwards JL. Heat-induced increases in body temperature in lactating dairy cows: impact on the cumulus and granulosa cell transcriptome of the periovulatory follicle. J Anim Sci. 2022. 1 (7): skac121. doi: 10.1093/jas/skac121   Lengi AJ, Stewart JW, Makris M, Rhoads ML, Corl BA. Heat stress increases mammary epithelial cells and reduces viable immune cells in milk of dairy cows. Animals (Basel). 2022. 12(20):2810. doi: 10.3390/ani12202810. PMID: 36290196; PMCID: PMC9597744.   Li Q, Chen KC, Bridges PJ, Matthews JC.  Pituitary and liver selenoproteins transcriptome profiles of grazing steers and their sensitivity to the form of selenium in vitamin-mineral mixes. Front Anim Sci.  2022. 3:911094. doi: 10.3389/fanim.2022.911094.   Lundberg AL, Jaskiewicz MN, Maucieri AM, Townson DH. Short Communication: Stimulatory Effects of TGFα in Granulosa Cells of Bovine Small Antral Follicles. J Anim Sci. 2022. 100:1-8. doi.org/10.1093/jas/skac105.   Maia TS, Guimarães HR, Garza V, Pohler KG, Cardoso RC, Williams GL. Early juvenile but not mid-to-late prenatal nutrition controls puberty in heifers but neither impact adult reproductive function. Biol Reprod. 2022 107(4):1035-45, 2022.   Mathew DJ, Peterson KD, Senn LK, Oliver MA, Ealy AD. Ruminant conceptus-maternal interactions: interferon-tau and beyond. J Anim Sci. 2022 Jul 1;100(7):skac123. doi: 10.1093/jas/skac123.   McCoski SR, Cockrum RR, Ealy AD. Short communication: Maternal obesity alters ovine endometrial gene expression during peri-implantation development. J Anim. Sci. 2022 Jul 1;100(7):skac090. doi: 10.1093/jas/skac090.   Moorey SE, Hessock EA, Edwards JL. Preovulatory follicle contributions to oocyte competence: Importance of the ever-evolving intrafollicular environment leading up to the LH surge. J Anim Sci. 2022. 1 (7): skac153. doi:10.1093/jas/skac153.   Moorey SE, Perry GA, Smith MF. Management Decisions to improve the reproductive performance of your herd: From calving to rebreeding. Applied Reproductive Strategies in Beef Cattle. 2022. San Antonio, TX.   Read CC, Edwards JL, Schrick FN, Rhinehart JD, Payton RR, Campagna SR, Castro HF, Klabnik JL, Moorey SE. Preovulatory serum estradiol concentration is positively associated with oocyte ATP and follicular fluid metabolite abundance in lactating beef cattle. J Anim Sci. 2022. 1 (7): skac136. doi: 10.1093/jas/skac136.   Soffa DR, Stewart JW, Arneson AG, Dias NW, Mercadante VRG, Rhoads RP, Rhoads ML. Reproductive and lactational responses of multiparous dairy cattle to short-term postpartum chromium supplementation during the summer months. J Dairy Sci Comm. 2022. doi:  https://doi.org/10.3168/jdsc.2022-0287.   Speckhart SL, Wooldridge LK, Ealy AD. An updated protocol for in vitro bovine embryo production. STAR Protoc. 2022 Dec 13;4(1):101924. doi: 10.1016/j.xpro.2022.101924.   Stewart JW, Arneson AG, Byrd MKH, Negron-Perez VM, Newberne HM, White RR, El-Kadi SW, Ealy AD, Rhoads RP, Rhoads ML. Comparison of production-related responses to hyperinsulinemia and hypoglycemia induced by clamp procedures or heat stress of lactating dairy cattle. J Dairy Sci. 2022 Oct;105(10):8439-8453. doi: 10.3168/jds.2022-21922.   Stoecklein KS, García-Guerra A, Duran BJ, Prather RS, Ortega M. Actions of FGF2, LIF, and IGF1 on Bovine Embryo Survival and Conceptus Elongation following Slow-rate Freezing. Front Anim Sci. 3. 2022. DOI: 10.3389/fanim.2022.1040064.   Walker BN, Nix J, Wilson C, Marrella MA, Speckhart SL, Wooldridge L, Yen CN, Bodmer JS, Kirkpatrick LT, Moorey SE, Gerrard DE, Ealy AD, Biase FH. Tight gene co-expression in BCB positive cattle oocytes and their surrounding cumulus cells. Reprod Biol Endocrinol. 2022 Aug 13;20(1):119. doi: 10.1186/s12958-022-00994-3.   Webb KL, Trotta RJ, Jia Y, Bridges PJ, Matthews JC. Influence of form of selenium supplementation and tall fescue toxicity on growth performance, serum parameters, and tissue mass of grazing beef steers.  Transl Anim Sci. 2022. 6(4):txac122. doi: 10.1093/tas/txac122.    Wooldridge LK, Keane JA, Rhoads ML, Ealy AD. Bioactive supplements influencing bovine in vitro embryo development. J Anim Sci. 2022 Jul 1;100(7):skac091. doi: 10.1093/jas/skac091.   Zhang M, Current JZ, Chaney HL, Yao J. Identification of the DNA binding element of ZNFO, an oocyte-specific zinc finger transcription factor in cattle. Gene. 2022 Aug 5;834:146655. doi: 10.1016/j.gene.2022.146655.   Rial C, Laplacette AL, Giordano JO. 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. 2022. 105:8411-8425. https://doi:10.3168/jds.2022-22082    Giordano JO, Sitko EM, Rial C, Perez MM, Granados GE. Use of multiple biological, management, and performance data for the design of targeted reproductive management strategies for dairy cows. J. Dairy Sci. 2022. 105:4669-4678. https://doi.org/10.3168/jds.2021-21476       Theses/Dissertations   Sarah N. Carr. Selenium form-induced changes in the early luteal phase corpus luteum, the blood, and the endometrium during early gestation in beef cows. PhD Dissertation. Kentucky.   Cassidy Ficker. Characteristics of Small Antral Follicles in Heifers Carrying the Bovine High Fecundity Allele Trio. Undergraduate Honors thesis. Ohio.   CB Hayden. Optimization of ovarian superstimulation before ovum pick-up and in vitro embryo production in pregnant cattle. MS thesis. Ohio.   Emma Hessock. Differential metabolome profiles in preovulatory follicular fluid of cattle with varying duration of proestrus. MS Thesis. Tennessee.   Emma Horn. Preovulatory follicular fluid and serum metabolome profiles in lactating beef cows with thin, moderate, and obese body condition. MS Thesis. Tennessee.   Ruben Lopez-Carrillo Jr. Ovarian protein hormones as biomarkers of fertility in dairy cows – Is it an acceptable model to predict infertility in dairy breeds? BS Thesis. Oregon.    Endya McKinley. Investigating the supplementation of IL-6, IL-11, & LIF at in vitro maturation to improve oocyte competency. MS Thesis. Virginia.   Katie Peterson. Identification of Endometrial Transcripts and Proteins Associated with the in vivo derived and in vitro produced bovine conceptus microenvironment. MS Thesis. Tennessee.   Casey Read. Effects of preovulatory follicle physiological status on oocyte metabolic capacity. PhD Dissertation. Tennessee.   Dallas Soffa  Effects of feed additives on uterine morphology and selected reproductive attributes. MS Thesis. Virginia.   Savannah L. Speckhart. Interleukin-6 and its Contribution to Embryogenesis in Cattle. PhD Dissertation. Virginia.   Emily Sitko. Genomically enhanced predictions of dairy cattle fertility: implementation in reproductive management and associations with cow reproductive biology and behavior. PhD Dissertation. Cornell University

Impact Statements

Specific impactful discoveries from the group include, but are not limited to: 1) describing the roles of pro-inflammatory cytokines and environmental stressors on follicle growth, steroidogenesis, luteal regression, and conceptus-uterine-immune interactions, 2) use of trace mineral supplementation to improve oocyte quality and increase systemic progesterone concentrations, 3) impact of beef cow body condition score and weight on the metabolome of follicular fluid and serum, 4) genetic determinants of ovulation and implantation rates, 5) endocrine and molecular mechanisms controlling ovulation rate in cows, 6) mechanisms associated with preimplantation embryo development and quality, 7) elucidate the underlying biology driving differences in dairy cow performance due to genetics, health, management, and environmental conditions, 8) developed novel reproductive management strategies that improve reproductive performance in dairy and beef herds, and 9) factors that cause pregnancy loss in cattle and development of methods to reduce losses. These findings were used to develop programs to improve synchronization of ovulation and to overcome environmental and nutritional factors that can disrupt reproduction. The group developed new management strategies and technologies to effectively use artificial insemination (AI) and embryo transfer (ET) technologies to improve reproductive performance and thereby immediately impact producers. Indeed, veterinarians, consultants, pharmaceutical companies, breed organizations, and companies serving animal industries benefitted from the work from this project. For example, collaborators have presented annual reports on the project to cattle AI organizations (Select Sires, Genex CRI, Alta Genetics), the National Association of Animal Breeders (NAAB), the American Association of Bovine Practitioners (AABP), dairy industry associations (Minnesota Dairy Association, Vermont Dairy Association), pharmaceutical and technology companies (Zoetis, Merck, Allflex), and extension education organization (PRO-Dairy). In turn, those groups spread the technology to farm families/producers for implementation which benefits the on-farm profitability and sustains agricultural production systems that are highly competitive in the global economy. Student training is another important impact of this project. Members of the NE227 group (NY, IA, MS, OH, OR, PA, KY, MA, NE, TN, VT, WI, VA) developed a course on Contemporary Topics in Reproductive Biology to improve student understanding of the breadth of reproductive physiology including topics outside their primary area of research. Greater than 50 students have enrolled in each of the last two offerings of the course. This accounted for a majority of the MS and PhD students across the experiment stations in this multistate project. Numerous undergraduate students with an interest in reproductive physiology were also introduced to investigative research. These activities represent an important contribution of the project to the education of the next generation of scientists, consultants, and other workers in animal agriculture industries.

Date of Annual Report: 09/30/2024

Report Information

Annual Meeting Dates: 05/14/2024 - 05/15/2024
Period the Report Covers: 10/01/2023 - 09/30/2024

Participants

Participants
Dave Townson
Cecily Bishop
Sarah Moorey
Dan Mathew
Soon Hun Cheong
Jen Wood
Phil Bridges
Joy Pate
Camilla Hughes
Alvaro Garcia Guerra
Aileen Keating
Joanne Fortune
Emelia Przygrodzka
Angela Gonella-Diaza
Julio Giordano
Alan Ealy
Vimal Selvaraj
Rodolfo Cardoso
Donnelly Hutchings
Paul Tsang
Francisco Diaz
Michaela Kuzniar

Brief Summary of Minutes

Accomplishments

Publications

Peer-reviewed journal articles reporting research from this project in 2023-2024 Hughes CHK, Smith OE, Meinsohn MC, Brunelle M, Gévry N, Murphy BD. Steroidogenic factor 1 (SF-1; Nr5a1) regulates the formation of the ovarian reserve. Proceedings of the National Academy of Sciences 2023; 120:e2220849120. https://doi.org/10.1073/pnas.2220849120 Pate JL, Hughes CHK. Luteal prostaglandins: mechanisms regulating luteal survival and demise in ruminants. Animal 2023; 17:100739. 10.1016/j.animal.2023.100739 Hughes CHK and Murphy BD. Steroidogenic factor 1 (SF-1; Nr5a1) orchestrates the establishment and depletion of the ovarian reserve. Society for the Study of Reproduction, 56th annual meeting. July 11-14, 2023, Ottawa, Ontario, Canada. (Invited) Krause, ART, Garner, TB and Diaz FJ. Inhibition of mouse oocyte maturation by a specific IK1 inhibitor. Society for the Study of Reproduction, July 11-14, 2023, Ottawa, Canada. Carothers A and Diaz FJ. Hormonal Regulation and Zinc Influx Activity of Zrt- and Irt-like Protein 9 (ZIP9) in the Mammalian Ovary. Society for the Study of Reproduction, July 11-14, 2023, Ottawa, Canada. Snider AP, Spuri-Gomes R, Summers AF, Tenley SC, Abedal-Majed MA, McFee RM, Wood JR, Davis JS, and Cupp AS (2023) Identification of lipids and cytokines in plasma and follicular fluid before and after FSH stimulation as potential markers for follicular maturation in cattle. Animals, 13:3289, PMID: 37894013 Ermisch AF, Bidne KL, Kurz SG, Bochantin KA, and Wood JR (2023) Ovarian inflammation mediated by TLR-4 increased transcripts of maternal effect genes and decreased embryo development. Biol Reprod 108(3): 423-436, PMID: 36461933 Ermisch AF and Wood JR (2023) N6-methyladenosine dynamics and differential methylation of maternal and zygotic mRNAs during the early stages of the maternal to zygotic transition. 56th Annual Meeting of the Society for the Study of Reproduction, Ottawa, Canada, Pre-doctoral Platform Competition Talk Monteiro P. L. J., Consentini C. E. C., Andrade J. P. N., Beard A. D., García-Guerra A., Sartori R., Wiltbank M. C. 2023. Research on timed AI in Beef Cattle: Past, Present and Future, a 27-year Perspective. Theriogenology, 211: 161-171. DOI: 10.1016/j.theriogenology.2023.07.037. Hayden C. B., Sala R. V., Pereira D. C., Moreno J. F., García-Guerra A. Use and dosage of p-FSH for ovarian superstimulation before ovum-pick up and in vitro embryo production in pregnant Holstein heifers. Journal of Diary Science, 106: 8110-8121. DOI: 10.3168/jds.2023-23576. Carranza Martin A, García-Guerra A, Relling A. 2023. Effects of polyunsaturated fatty acid supplementation on plasma and follicular fluid Resolvin D1 concentration and the expression of mRNA of genes associated with resolving synthesis in ewes. Journal of Animal Science, 101: skad310.  DOI: 10.1093/jas/skad310. Constantino, J. V., Carranza-Martin A., Premanandan C., Kirkpatrick B. W., Wiltbank M. C., García-Guerra A. 2023. Preantral follicle numbers and size in heifers carrying Trio, a bovine high fecundity allele. Reproduction, 166: 13-26. DOI: 10.1530/REP-23-0072. Rojas Canadas E., Battista S.E., Kieffer J.D., Wellert S.R., Mussard M. L., Garcia-Guerra A. 2023. GnRH dose at initiation of a 5-day CO-Synch + P4 for fixed time artificial insemination in suckled beef cows. Animal Reproduction Science, 250: 107210.  DOI: 10.1016/j.anireprosci.2023.107210. <ol start="2023"> <li>Rykaczewski, N. B. Bello, J. D. Kieffer, A. Z. Chang, M. Trotter, A. Garcia-Guerra. 2023. “Evaluation of on-animal sensors for monitoring diurnal physical activity of beef bulls during the breeding season” Journal of Animal Sciences 101(Suppl. 3): 545-546. DOI: 10.1093/jas/skad281.641</li> </ol> <ol start="386"> <li>P. Folchini, B. J. Duran, C. Rykaczewski, A. E. Crist, M. Saad, A. C. Carranza-Martin, J. C. L. Motta, D. F. Mollenkopf, M. L. Mussard, A. Garcia-Guerra. “Induction of endometritis after pregnancy loss during the second month of gestation in cattle and its effect on the fate of the corpus luteum” Proceedings of the 56th Annual Meeting of the Society for the Study of Reproduction 2023, July 11th – 14th, Ottawa, Canada, Page 386.</li> </ol> <ol start="380"> <li>F. Incarnato, A. E. Crist, J. C. L. Motta, C. B. Hayden, A. Garcia-Guerra. “Assessment of circulating anti-Müllerian hormone during pregnancy in beef cattle” Proceedings of the 56th Annual Meeting of the Society for the Study of Reproduction, July 11th – 14th, 2023, Ottawa, Canada, Page 380.</li> </ol> <ol start="2023"> <li>C. L. Motta, R. V. Sala, C. Hayden, D. C. Pereira, P. J. Ross, J.F. Moreno, A. Garcia-Guerra. 2023. “Ovarian stimulation with FSH increases in vitro embryo production in high AMH heifers in a dose-dependent manner”. Animal – science proceedings 14(3): 492. DOI: 10.1016/j.anscip.2023.03.099</li> </ol> <ol start="2023"> <li>Carranza-Martin, C. Furnus, A. García-Guerra, A. Relling. 2023. “Effects of polyunsaturated fatty acids supplementation in resolvin D1 concentration in plasma and follicular fluid in ewes”. Journal of Animal Sciences 101(Suppl. 2): 332-333. DOI: 10.1093/jas/skad341.378</li> </ol> <ol start="2023"> <li>B. Hayden, J. Motta, N. Bello, R. Sala, V. Absalon-Medina, P. Ross, J. Moreno, A. Garcia-Guerra. 2023. “Assessment of methods to synchronise follicle wave emergence in pregnant heifers”. Reproduction Fertility and Development 35(2):204-205. DOI: 10.1071/RDv35n2Ab153.</li> </ol> Hutchings DW, Elder D, Tsang PCW 2024. Effects of Phytoestrogens on Progesterone Biosynthesis in Dairy Cows. Poster; ASAS-CSAS-WSASAS Annual Meeting July 2024, Calgary, Canada. Maucieri AM, Townson DH 2024. Influence of O-GlcNAcylation on KGN cell function and metabolism. Endocrine and Metabolic Science (In Revision) Alward KJ, Cockrum RR, Ealy AD. Associations of antral follicle count with fertility in cattle: A review. JDS Commun. 2023 Jan 31;4(2):132-137. doi: 10.3168/jdsc.2022-0283. Nix J, Marrella MA, Oliver MA, Rhoads M, Ealy AD, Biase FH. Cleavage kinetics is a better indicator of embryonic developmental competency than brilliant cresyl blue staining of oocytes. Anim Reprod Sci. 2023 Jan;248:107174. doi: 10.1016/j.anireprosci.2022.107174. Nix JL, Schettini GP, Speckhart SL, Ealy AD, Biase FH. Ablation of OCT4 function in cattle embryos by double electroporation of CRISPR-Cas for DNA and RNA targeting (CRISPR-DART). PNAS Nexus. 2023 Oct 20;2(11):pgad343. doi: 10.1093/pnasnexus/pgad343. Rhoads ML. Review: Reproductive consequences of whole-body adaptations of dairy cattle to heat stress. Animal. 2023 May;17 Suppl 1:100847. doi: 10.1016/j.animal.2023.100847. Soffa, D.R., J.W. Stewart, E.D. Pack, A.G. Arneson, R. De Vita, J.W. Knight, D.W. Fausnacht, R.P. Rhoads, S.G. Clark, D.G. Schmale and M.L. Rhoads. 2023. Short-term consumption of the mycotoxin zearalenone by pubertal gilts causes persistent changes in the histoarchitecture of reproductive tissues. J Anim Sci. 2023 Jan 3;101:skac421. doi: 10.1093/jas/skac421. PMID: 36574505. Speckhart SL, Oliver MA, Ealy AD. Developmental Hurdles That Can Compromise Pregnancy during the First Month of Gestation in Cattle. Animals (Basel). 2023 May 25;13(11):1760. doi: 10.3390/ani13111760. Speckhart SL, Wooldridge LK, Ealy AD.  An updated protocol for in vitro bovine embryo production. STAR Protoc. 2023 Mar 17;4(1):101924. doi: 10.1016/j.xpro.2022.101924. Arneson AG, Stewart JW, Byrd MH, Perry GA, Rhoads ML. Plasma γ-Aminobutyric Acid (GABA) Concentrations in Lactating Holstein Cows during Thermoneutral and Heat Stress Conditions and Their Relationships with Circulating Glucose, Insulin and Progesterone Levels. Vet Sci. 2024 Mar 21;11(3):137. doi: 10.3390/vetsci11030137. Keane JA, Ealy AD. An Overview of Reactive Oxygen Species Damage Occurring during In Vitro Bovine Oocyte and Embryo Development and the Efficacy of Antioxidant Use to Limit These Adverse Effects. Animals (Basel). 2024 Jan 21;14(2):330. doi: 10.3390/ani14020330. McKinley E, Speckhart SL, Keane JA, Oliver MA, Rhoads ML, Edwards JL, Biase FH, Ealy AD.  Influences of Supplementing Selective Members of the Interleukin-6 Cytokine Family on Bovine Oocyte Competency. Animals (Basel). 2023 Dec 21;14(1):44. doi: 10.3390/ani14010044.   Oliver MA, Speckhart SL, Ealy AD. Interleukin-6 Family Members Improve Pre-Implantation Bovine Blastocyst Composition. 2023 American Association of Animal Science Meeting, Albuquerque, NM. Oliver MA, Speckhart SL, Edwards JL, Ealy AD. Human Recombinant Interleukin-6 has Cryoprotective Properties in Preimplantation Bovine Embryos. 2023 Society for the Study of Reproduction Annual Meeting, Ottawa, ON, CA.   McKinley E, Speckhart SL, Keane JA, Oliver MA, Ealy AD. Supplementing IL6, IL11, and LIF to improve culture bovine oocyte competency. 2023 Society for the Study of Reproduction Annual Meeting, Ottawa, ON, CA.   Nix JL, Speckhart SL, Schettini GP, Ealy AD, Biase FH. Disruption of OCT4 in cattle embryos by electroporation of CRISPR-Cas ribonucleoproteins into zygotes for DNA and RNA targeting (CRISPR-DART). 2023 Society for the Study of Reproduction Annual Meeting, Ottawa, ON, CA.   Speckhart SL, Oliver MA, Keane JA, Dias NW, Mercadante VRG, Biase FH, Ealy AD. Interleukin-6 control of conceptus elongation and survival is multifaceted in the cow. 2023 Society for the Study of Reproduction Annual Meeting, Ottawa, ON, CA.   Keane JA, Wooldridge LK, Speckhart SL, Nix JL, Biase FH, Ealy AD. Modifications in media composition supports increased blastocyst development and inner cell mass numbers of in vitro produced bovine embryos. 2023 Society for the Study of Reproduction Annual Meeting, Ottawa, ON, CA.   Habeeb, H. M. H., Kleditz, L., Hazzard, T. M., Bishop, C. V., Stormshak, F., Kutzler, M. A. (2023). Ovine endometrial estrogen receptor expression is altered following PG-600 administration. Veterinary Medicine and Science. 9(3). DOI:10.1002/vms3.1119 Kim, S.-Y., Xu, J., Mayerhofer, A., Bishop, C. V. (2023). Editorial: Small molecules and peptides in paracrine/autocrine regulation of ovarian folliculogenesis. Frontiers in Endocrinology, 14. DOI:10.3389/fendo.2023.1168701 Slayden, O., Bishop, C. V. The Primate Uterus: Cyclic Changes Encyclopedia of Reproduction (3rd Edition). Accepted for Publication November 2023. Elsevier. Bishop, C., Stricklin, O., Fox, A., Harrington, S., Estill, C., Zelinski, M., Ting, A. “Ovarian activity after autologous cryopreservation and transplantation in ewes.” Poster Presentation, 56th Annual Meeting, Society for the Study of Reproduction, July 11-14 2023, Ottawa, Canada. Carr, S.N., B.R. Crites, H. Shinde and P.J. Bridges.  2023.  Transcriptomic changes in response to form of selenium on the interferon-tau signaling mechanism in caruncular tissue of beef heifers at maternal recognition of pregnancy.  International Journal of Molecular Sciences.  24:17327. doi:10.3390/ijms242417327. S.N. Carr, B.R. Crites and P.J. Bridges.  2023.  Effects of form of selenium on serum metabolites and iodothyronine deiodinases during early gestation in beef heifers.  Special Issue, Proc. of the 56th annual meeting of the Soc. for the Study of Reproduction. Rial, C., Hussain, I., Hoff, R., Tompkins, S., Erickson, D., Branen, J. and Giordano, J.O.* 2024. Development and evaluation of a lateral flow-based portable optical system for determination of the pregnancy status of dairy cows. J. Dairy Sci.  Online ahead of print. <a href="https://doi.org/10.3168/jds.2024-24899">https://doi.org/10.3168/jds.2024-24899</a> Laplacette A. L., Rial C., Magaña Baños G. S., García Escalera J. S., Torres S., Kerwin A., and Giordano J. O.* 2024. Effect of a targeted reproductive management program based on automated detection of estrus during the voluntary waiting period on reproductive performance of lactating dairy cows. Theriogenology 225:130-141. <a href="https://10.1016/j.theriogenology.2024.05.030">https://10.1016/j.theriogenology.2024.05.030</a>      Sitko E. M., Laplacette A. L., Duhatschek D., Rial C., Perez M. M., Tompkins S., Kerwin A. L., Domingues R. R., Wiltbank  M. C. and Giordano J. O.* 2023. Ovarian function and endocrine phenotypes of lactating dairy cows during the estrous cycle are associated with genomic-enhanced predictions of fertility potential. J. Dairy Sci. 107:7352-7370. <a href="https://10.3168/jds.2023-24378">https://10.3168/jds.2023-24378</a>   Sitko E. M., Laplacette A. L., Duhatschek D., Rial C., Perez M. M., Tompkins S., Kerwin A. L.  and Giordano J. O.* 2023. Reproductive physiological outcomes of lactating dairy cows with different genomic merit for fertility: biomarkers, uterine health, endocrine status, estrus features, and response to synchronization of estrus and ovulation. J. Dairy Sci. <a href="https://10.3168/jds.2023-24376">https://10.3168/jds.2023-24376</a> Online ahead of print. Rial C., and J. O. Giordano*. 2023. Combining reproductive outcomes predictors and automated estrus alerts recorded during the voluntary waiting period identified subgroups of cows with different reproductive performance potential. J. Dairy Sci. 107:7299-7316. <a href="https://10.3168/jds.2023-24309">https://10.3168/jds.2023-24309</a> Laplacette A. L., Rial C., Duhastchek D., Perez M.M., Stangaferro M. L., Thomas M. J. and Giordano J. O.* 2023. Effect of delaying induction of ovulation on ovarian function of lactating dairy cows synchronized with Double-Ovsynch and inseminated after detected estrus or timed AI. J. Dairy Sci. 106 (E-Suppl. 1). Rial C., Stangaferro M. L., Thomas M. J. and Giordano J. O.* 2023. Association between vaginal discharge scores with rumination time, activity time, a health index score, and milk yield in lactating dairy cows. J. Dairy Sci. 106 (E-Suppl. 1). Rial C., Hussain I., Erickson D., Branen J. and Giordano J. O.* 2023. Development and demonstration of lateral-flow immunoassays for determination of the pregnancy and ovarian physiological status of cows. J. Dairy Sci. 106 (E-Suppl. 1). Theses/Dissertations IMMUNOLOGICAL AND ENDOMETRIAL CHANGES OF EARLY PREGNANCY IN DAIRY CATTLE. 2023. Doctoral Dissertation in Integrative and Biomedical Physiology. Maria Isabel da Silva, Pennsylvania State University. DeCastro, LN (2024) Evaluation of the effects of O-GlcNAcylation and Hypoxia on Estradiol Production by Bovine Granulosa Cells. URI MS Thesis Endya McKinley (2023). Investigating the supplementation of IL-6, IL-11, & LIF at in vitro maturation to improve oocyte competency (MS Thesis). Savannah L. Speckhart (2023).  Interleukin-6 and its Contribution to Embryogenesis in Cattle (PhD Dissertation). Carr, Sarah Nancy, "SELENIUM FORM-INDUCED CHANGES IN THE EARLY LUTEAL PHASE CORPUS LUTEUM, THE BLOOD, AND THE ENDOMETRIUM DURING EARLY GESTATION IN BEEF COWS" (2023). Theses and Dissertations--Animal and Food Sciences. 138. https://uknowledge.uky.edu/animalsci_etds/138

Impact Statements

Specific impactful discoveries from the group include, but are not limited to: 1) describing the roles of pro-inflammatory cytokines and environmental stressors on follicle growth, steroidogenesis, luteal regression, and conceptus-uterine-immune interactions, 2) use of trace mineral supplementation to improve oocyte quality and increase systemic progesterone concentrations, 3) impact of beef cow body condition score and weight on the metabolome of follicular fluid and serum, 4) genetic determinants of ovulation and implantation rates, 5) endocrine and molecular mechanisms controlling ovulation rate in cows, 6) mechanisms associated with preimplantation embryo development and quality, 7) elucidate the underlying biology driving differences in dairy cow performance due to genetics, health, management, and environmental conditions, 8) developed novel reproductive management strategies that improve reproductive performance in dairy and beef herds, and 9) factors that cause pregnancy loss in cattle and development of methods to reduce losses. These findings were used to develop programs to improve synchronization of ovulation and to overcome environmental and nutritional factors that can disrupt reproduction.
The group developed new management strategies and technologies to effectively use artificial insemination (AI) and embryo transfer (ET) technologies to improve reproductive performance and thereby immediately impact producers.
collaborators have presented annual reports on the project to cattle AI organizations (Select Sires, Genex CRI, Alta Genetics), the National Association of Animal Breeders (NAAB), the American Association of Bovine Practitioners (AABP), dairy industry associations (Minnesota Dairy Association, Vermont Dairy Association), pharmaceutical and technology companies (Zoetis, Merck, Allflex), and extension education organization (PRO-Dairy). In turn, those groups spread the technology to farm families/producers for implementation which benefits the on-farm profitability and sustains agricultural production systems that are highly competitive in the global economy.
Student training is another important impact of this project. Members of the NE227 group (NY, IA, MS, OH, OR, PA, KY, MA, NE, TN, VT, WI, VA) developed a course on Contemporary Topics in Reproductive Biology to improve student understanding of the breadth of reproductive physiology including topics outside their primary area of research. More than 50 students have enrolled in each of the last two offerings of the course. This accounted for a majority of the MS and PhD students across the experiment stations in this multistate project. Numerous undergraduate students with an interest in reproductive physiology were also introduced to investigative research. These activities represent an important contribution of the project to the education of the next generation of scientists, consultants, and other workers in animal agriculture industries.

Date of Annual Report: 06/05/2025

Report Information

Annual Meeting Dates: 05/12/2025 - 05/14/2025
Period the Report Covers: 10/01/2024 - 09/30/2025

Participants

Cecily Bishop (Oregon)
Phil Bridges (Kentucky)
Rodolfo Cardoso (Texas)
Alan Ealy (Virginia)
Maria Gracia Gervasi (Connecticut)
Julio Giordano (Cornell)
Angela Gonella (Florida)
Alvaro Garcia-Guerra (Ohio)
Camilla Hughes (Pennsylvania)
Aileen Keating (Iowa)
Dan Mathews (Tennessee)
Jessica Motto (Maine)
Amanda Patterson (Missouri)
Shelly Rhoads (Virginia)
Cecilia Rocha (Missouri)
Kosta Siminitras (Louisiana)
Dave Townson (Rhode Island)
Milo Wiltbank (Wisconsin)
Jen Wood (Nebraska)

Brief Summary of Minutes

Brief Summary of Minutes of Annual Meeting

Host: Dr. Alvaro Garcia-Guerra, The Ohio State University

Monday, May 12^th, 2025

Arrive at the Deer Creek Lodge

Dinner on own

Tuesday May 13^th, 2024

09:00 Call to order; Alan Ealy, Virginia Tech

Introductions

Group business

Meeting plan

9:15 Short presentations by new members:

Maria Gracia Gervasi (Connecticut)

Cecilia Rocha (Missouri)

Jessica Motto (Maine)

9:30 Alvaro Garcia-Guerra (Ohio)

9:45 Aileen Keating (Iowa)

10:00 Rodolfo Cardoso (Texas)

10:15 Break

10:30 Kamilah Grant and Mark Mirando

USDA-NIFA Updates

11:00 Jen Wood (Nebraska)

11:15 Angela Gonella (Florida)

11:30 Julio Giordono (Cornell)

11:45 Milo Wiltbank (Wisconsin)

12:00 Lunch

1:00 Amanda Patterson (Missouri)

1:15 Shelly Rhoads (Virginia)

1:30 Dan Mathews (Tennessee)

1:45 Camilla Hughes (Pennsylvania)

2:00 Leave for Tour of ST Genetics

11000 Huntington Rd

South Charleston, OH

Group Dinner-to be determined

Wednesday May 14^th, 2025

9:00 Cecily Bishop (Oregon)

9:15 Kosta Siminitras (Louisiana)

9:30 Alan Ealy (Virginia)

9:45 Graduate Reproductive Biology Course Update

Alan Ealy and others

10:00 New Collaborative Teaching Opportunites

Dave Townson and Alan Ealy

10:30 Break out groups to discuss collaborative research and teaching opportunities

11:15 Reports of collaborative efforts that were discussed

12:00 Adjourn

Accomplishments

Accomplishments Objective 1: Identify Mechanisms that Regulate Ovarian Function and Oocyte Quality during the Estrous Cycle Determined the effect of Ang II on CCN1 expression, and the activity of gelatinase MMPs (MMP2 and MMP9), in steroidogenic cells obtained from the mid-cycle bovine corpus luteum. Determined the effect of calcium ionophore on CCN1 expression, and the activity of gelatinase MMPs (MMP2 and MMP9) by steroidogenic cells obtained from the mid-cycle bovine corpus luteum. Determined the effects of phytoestrogens in red clover on progesterone (P4) and prostaglandin F2alpha (PGF2α) production in bovine corpora lutea. Proteomic profiling revealed potential mechanisms regulating prostaglandin F2α responsiveness in bovine corpora lutea A role of Zinc Insufficiency has been identified in Bovine Granulosa Cell Viability. A recent set of studies suggests that most of the temporal changes in the CL during early pregnancy that we previously reported are not regulated directly by IFNT, drawing into question identities of other luteal regulators during early pregnancy or the effect of age of CL independent of embryonic signaling. Recent work described the effect of FSH dose on ovarian response and in vitro embryo production in pregnant cattle with low AMH. Generated new knowledge related to the impact of preovulatory follicle maturity on oocyte metabolism and competence for embryo development based on metabolome data of maturing oocytes, in vitro embryo development, and RNA-sequencing results of blastocysts.  Demonstrated that circulating and follicular fluid metabolites are differentially regulated in lactating beef cows of thin, obese, and moderate body composition, and that systemic regulation of reactive oxygen species may be dependent on body condition score. Explored the extent to which higher estrus associated body temperature advanced preovulatory follicle progression and the metabolome of the preovulatory follicle.  New knowledge was also generated pertaining to the impact of elevated body temperature induced by an acute episode of heat stress after the LH surge impacts preovulatory follicle metabolome.  Recent work showed that increased DNA damage in oocytes is correlated with diminished autophagy activation. Heat stress impacts insulin and glucose in dairy cattle experiencing heat stress. This adversely influences preovulatory follicular environment to oocyte quality and competency for embryonic development.  Studies have described the relative contributions of steroid hormones as well as undefined components of the follicular fluid. An improvement in fertility is observed following displayed estrus in a fixed-time artificial insemination protocol.  Follicular fluid contributes to fertility under these circumstances as blastocyst development was greater when oocytes were exposed to follicular fluid from animals that did display estrus. Determined that KGN cells, an immortal granulosa cell tumor line, express proteins that are post translationally modified by O-GlcNAcylation and that manipulation of O-GlcNAcylation in vitro impairs KGN proliferation and oxidative phosphorylation. Demonstrated that manipulation of O-GlcNAcylation in primary cultures of bovine granulosa cells impairs transcript expression of STAR but otherwise has no effect on expression of FSHR and CYP19A1 transcripts, or estradiol production. Provided preliminary evidence that induction of hypoxia in primary cultures of bovine granulosa cells augments estradiol production; however, manipulation of O-GlcNAcylation does not impact this. This work also identified 354 proteins of the granulosa cell proteome are putatively modified by O-GlcNAcylation, and these proteins encompass a wide range of functional categories including transcription factors, metabolic enzymes, and cytoskeletal proteins. Identified the signaling pathways and epigenomic factors that regulate RNA binding protein-mRNA interactions during oocyte growth and maturation. We hypothesize that changes in oocyte signaling pathways, i.e., increased ERK1/2, AMPK, and CDK1, that occur after the LH surge, regulate the phosphorylation of BTG4, ELAVL1 ZFP36L2, and MSY2 and subsequently the binding affinity of each RBP to a target 3’UTR. Determined the effects of luteinizing hormone/protein kinase (PKA) signaling on metabolic pathways driving function of luteal cells and progesterone production. Explored the signaling and metabolic pathways regulating the function of granulosa cells. We determined the PKA-induced metabolic changes and content of related proteins as well as function of glucose metabolism branches on granulosa cell function. Examined the roles of metabolites as signaling molecules in the somatic ovarian cells. We found that main kinase triggered by gonadotropins, such as protein kinase A (PKA), enhances histone acetylation and increases chromatin availability in the granulosa cells.  Objective 2: Determine Factors Associated with Fertilization, Embryo Development, and Conceptus-Endometrial Interactions that Dictate Pregnancy Success Investigated the role of methyl donors during the periconceptional period and their influence on embryonic development, pregnancy establishment, offspring development, and epigenetic modifications such as DNA methylation. Ongoing research is evaluating how rumen-protected methionine (RPM) supplementation during the periconceptional period (7 days before to 7 days after timed artificial insemination) affects postnatal development of offspring. Additional studies are exploring the impact of RPM supplementation in oocyte donor cows on the composition of follicular fluid and its potential to enhance in vitro bovine embryo production. Described impacts of environmental exposures on the function of the ovary. Protein profiling of those changes within the ovary are now being associated with loss of ovarian function, changes to hormone production, and impairments to the female gamete. Explored the functional role of endometrial prostaglandin F₂α in luteal regression after conceptus demise in cattle. Uncovered the “uterine secretory cycle” as a physiological determinant of preimplantation success. Through temporal transcriptomic profiling of endometrial secretions across the estrous cycle, the work established a foundational framework which governs the cyclical preparedness of the uterus for sperm transit and early embryo development. This highlights how hormone-driven endometrial outputs align precisely with fertilization and preimplantation requirements. It was revealed that there is a limited responsiveness of the endometrium to embryos during the cycle. The uterine environment is pre-programmed rather than reactively modulated in response to embryos, refining the understanding of when conceptus-endometrial interactions are most critical and sensitive for successful implantation. Linked uterine luminal conditions to metabolic support of embryo elongation. The identification of a quasi-anoxic uterine environment that favors anaerobic glycolysis provides mechanistic insight into how the uterus metabolically supports rapid conceptus expansion, a critical feature for implantation in Ruminantia. Completed oral dosing pilot study to determine if gestating beef cows consume enough vitamin A and D precursors via free choice mineral supplement to raise serum levels of vitamin A and D above threshold previously associated with achievement of pregnancy after first artificial insemination cycle post-calving.  Explored how the interleukin-6 (IL6) cytokine family embryonic disk development and conceptus elongation. Recent discoveries include that supplementing IL6 improves bovine conceptus elongation, that IL6 controls epiblast development, and that LIF preferentially controls hypoblast development. Antioxidant supplementation can improve in vitro bovine embryo production. Identified a conditioned media with the capacity to increase day 7 IVP embryo development by 60%. An increase in embryo quality was also observed. Described endometrial biological responses specific to IVP cattle embryos and IVP cattle embryo specific molecules that elicit these responses. Expanded vitamin A and D analyses to begin to evaluate effects of vitamin E alone or in combination with A and D on breeding success following first cycle artificial insemination. Future studies will evaluate A, D and E impacts on embryo development and chromosomal integrity.  Characterized adenomyosis (ectopic endometrial glands and stroma in the myometrium) in postpartum dairy cows and is investigating its effect on fertility. An association between postpartum metritis and the severity of adenomyosis was uncovered, including increased fibrosis, which may impact the receptivity of the uterus to an embryo. Investigated the impact of metritis/endometritis on postpartum endometrial epithelial regeneration and the effects on fertility. A putative endometrial epithelial stem/progenitor cell population has been identified, and this stem cell is being expanded in diseased endometrium in an attempt to repair the epithelial barrier. Investigated the feasibility of using endometrial cytology 14 days after artificial insemination without harming the pregnancy outcome. Collecting cytobrush samples on day 14 does not harm the outcome of the pregnancy, and harvested cells are luminal epithelial and glandular epithelial in nature.  Determined post-transcriptional mechanisms that regulate steroidogenesis by ovarian somatic cells. It is our hypothesis that regulation of steroidogenesis in ovarian granulosa and theca cells during the estrous cycle as well as the change in steroidogenesis during transition of granulosa and theca cells to luteal cells is dependent on changes in RNA methylation and the translation of the proteins that liberate and convert cholesterol to progesterone, androgen, and/or estrogen. Objective 3: Develop and Evaluate Novel Reproductive Management Strategies and Technologies to Improve Reproductive Performance of Ruminants Evaluated a novel recombinant hormone protocol for embryo production in Holstein heifers using a single injection of long-acting human follicle-stimulating hormone (FSH). Current investigations are assessing the use of low-dose human chorionic gonadotropin (hCG) at the time of progesterone device withdrawal to support follicular development. Optimization of synchronization protocols were pursued in an attempt to maximize fertility and reproductive efficiency. Determined the effect of a targeted reproductive management programs based on automated detection of estrus during the voluntary waiting period on reproductive performance of lactating dairy cows.  Continued to experiment with estrous synchronization protocols to define how many times progesterone devices can be reused without harming the fertility of the cow. Provided evidence that combining reproductive outcomes predictors and automated estrus alerts recorded during the voluntary waiting period identified subgroups of cows with different reproductive performance potential. Demonstrated that delayed induction of ovulation affects the ovarian function of lactating dairy cows synchronized with the Double-Ovsynch protocol and increases insemination after detected estrus. Developed novel reproductive management programs aimed at increasing fertility of first service through promotion of expression of estrus at the end of synchronization of ovulation protocols. Identified associations between milk production potential and the profitability of primiparous Holstein cows submitted to insemination with programs that prioritized insemination at detected estrus or timed artificial insemination Defined a lateral flow-based electronic system for determination of the pregnancy and ovarian status of cattle. Validated antibody detecting von Willebrand Factor to detect vascularization in sheep ovaries as part of ongoing efforts to study a sheep model of ovarian vitrification and re-transplantation in collaboration with 21st Century Medicine and Expanse Bio, LLC. Completed fertility analyses of an additional cohort of ewes in this ovarian vitrification model including performing in vitro fertilization of recovered oocytes from vitrified, warmed and re-transplanted ovaries. Identified novel candidate genes and proteins that may regulate luteal responsiveness to PGF2A Evaluated regulation of luteal gene expression by the maternal recognition of pregnancy signal, IFNT, and determined that the luteal gene expression changes that occur in early pregnancy are mostly not regulated by IFNT

Publications

Peer-reviewed journal articles (2024 Arneson AG, Stewart JW, Byrd MH, Perry GA, Rhoads ML. 2024. Plasma GABA concentrations in lactating Holstein cows during thermoneutral and heat stress conditions and their relationships with circulating glucose, insulin, and progesterone levels. Vet. Sci. 11:137. doi: 10.3390/vetsci11030137.  Azari-Dolatabad N, Stokes AE, Hessock EA, Edwards JL, Payton RR, Clark HM, Schrick FN, Moorey SE. 2024. Proestrus, follicle dynamics, and hormone profiles in Jersey cows during synchronization and re-synchronization after preovulatory follicle aspiration. Journal of Animal Science and Research. 8(1). doi:10.16966/2576-6457.167. Brochado C, Duran B J, Kieffer JD, Pinczak A, Menchaca A, Garcia-Guerra A. 2024. Reduced embryo yield obtained from superstimulated ewes with low ovarian follicular reserve is improved by lengthening the FSH treatment. Theriogenology, 218: 79-88. DOI: <a href="https://doi.org/10.1016/j.theriogenology.2024.01.024">10.1016/j.theriogenology.2024.01.024</a>. Cavalcante de Souza, D, Gonella-Diaza A, Tonizza de Carvalho NA, Elliff FM, Soares de Carvalho JG, Mendes Vieira L, Bonfim Neto AP, de Carvalho Papa P, Ghuman SS, Madureira EH, Pugliesi G, Binelli M, Baruselli PS 2024. Supplementation with Long-Acting Injectable Progesterone Three Days after TAI Impaired Luteal Function in Buffaloes. Tropical Animal Health and Production 56: article number 76. DOI: 10.1007/s11250-024-03913-3. Chang WJ, Baker MS, Laritsky E, Gunasekara CJ, Maduranga U, Galliou J, McFadden JW, Waltemyer JR, Berggren-Thomas B, Tate BN, Zhang HH, Rosen BD, Van Tassell CP, Liu GE, Coarfa C, Ren YA, Waterland RA 2024. Systemic interindividual epigenetic variants in cattle share major hallmarks with those in humans. uly 15th. Genome Biology. https://genomebiology. biomedcentral.com/articles/10.1186/s13059-024-03307-6 Clark HM, Stokes AE, Edwards JL, Payton RR, Schrick FN, Campagna SR, Sarumi Q, Hessock EA, Roberts SR, Azaridolatabad N, Moorey SE. 2024. Impact of preovulatory follicle maturity on oocyte metabolism and embryo development. PNAS Nexus. 3(5):pgae181. doi:10.1093/pnasnexus/pgae181. Ermisch AF and Wood JR. 2024. Regulation of Oocyte mRNA Metabolism: A Key Determinant of Oocyte Developmental Competence. Adv Anat Embryol Cell Biol, 238: 23-46, PMID: 39030353. Gonella-Diaza A, Sponchiado M, Rodrigues França M, Liu L, Pugliesi G, Guimarães Lo Turco E, Peñagaricano F, Binelli M. 2024. The metabolomic composition of the oviductal fluid is controlled by the periovulatory hormonal context in Bos indicus cows. Biology of Reproduction 111(6): 1188-1201. DOI: 10.1093/biolre/ioae153. González-Alvarez ME, Antwi-Boasiako C, Keating AF. 2024. Effects of per- and polyfluoroalkylated substances on female reproduction. Toxics. 12(7):455. doi: 10.3390/toxics12070455. González-Alvarez ME, Inyang I, Keating AF. 2024. Exposure to 7,12-dimethylbenz[a]anthracene impacts ovarian DNA damage sensing and repair proteins differently in lean and obese female mice and weight loss may mitigate obesity-induced ovarian dysfunction. Toxicology and Applied Pharmacology. 486:116930. doi: 10.1016/j.taap.2024.116930 Hayden C B, Motta JCL, Sala RV, Bello N, Coutinho da Silva M, García-Guerra A. 2024. Efficacy of methods to synchronize follicular wave emergence in pregnant heifers. JDS Communications, JDSC.2024-0629.R1, In press. DOI: <a href="https://doi.org/10.3168/jdsc.2024-0629">10.3168/jdsc.2024-0629</a>. Hill KS, Edwards JL, Payton RR, Schrick FN, Campagna SR, Hessock EA, Moorey SE. 2024. Relationships of circulating and preovulatory follicular fluid hydrogen peroxide level with body condition score and metabolome profiles of lactating beef cows. Agriculture. 14(8):1406. doi: 10.3390/agriculture14081406. Inyang I, White HE, Timme K, Keating AF. 2024. Biological sex differences in hepatic response to in utero dimethylbenz[a]anthracene exposure. Reproductive Toxicology. 124:108553. doi: 10.1016/j.reprotox.2024.108553. PMID: 38307155. Joyce K, Gad A, Menjivar NG, Gebremedhn S, Heredia D, Dubeux G, Lopez-Duarte MC, Bittar J, Gonella-Diaza A, Tesfaye D. 2024. Seasonal environmental fluctuations alter the transcriptome dynamics of oocytes and granulosa cells in beef cows. Journal of Ovarian Research 17(1): article number 201. DOI: 10.1186/s13048-024-01530-0. Keane JA, Ealy AD. 2024. An Overview of Reactive Oxygen Species Damage Occurring during In Vitro Bovine Oocyte and Embryo Development and the Efficacy of Antioxidant Use to Limit These Adverse Effects. Animals (Basel). Jan 21;14(2):330. doi: 10.3390/ani14020330. Keating AF, Ross JW, Baumgard LH. 2024. Systemic and Ovarian Impacts of Heat Stress in the Porcine Model. Reproduction. Nov 11;168(6):e240217. doi: 10.1530/REP-24-0217. Maucieri AM, Townson DH 2024. Influence of O-GlcNAcylation on KGN cell function. Endocrine and Metabolic Science 16:100192. Mirzaei A, Londoño-Mendez MC, Lasso-Ramirez S, Adams PE, Seekford ZK, Bromfield JJ, Heredia D, Di Lorenzo N, Chebel RC, Gonella-Diaza AM. 2024. Embryo production by Holstein heifers superovulated with a recombinant long-acting follicle stimulating hormone analog. Journal of Animal Sciences 102: skae326. DOI: 10.1093/jas/skae326. Mills MD, Pollock AB, Batey IE, O’Neil MA, Schrick FN, Payton RR, Moorey SE, Fioravanti P, Hipsher W, Zoca SM, Edwards JL. 2024. Magnitude and persistence of higher estrus associated temperatures in beef heifers and suckled cows. Journal of Animal Science. 102. Skae079. doi: 10.1093/jas/skae079. Novbatova G., Timme K, Keating AF. 2024. High fat diet-induced obesity and gestational DMBA exposure alter folliculogenesis and the proteome of the maternal ovary. Biology of Reproduction. 111(2):496-511. doi: 10.1093/biolre/ioae070. Laplacette AL, Rial C, Sitko EM, Perez MM, Tompkins S, Stangaferro ML, Thomas MJ, Giordano JO. 2024. Delaying induction of ovulation and timed artificial insemination in a Double-Ovsynch protocol increased expression of estrus and altered first-service reproductive outcomes of lactating dairy cows. J. Dairy Sci. 2024 Oct 14:S0022-0302(24)01216-5. <a href="https://doi:10.3168">https://doi:10.3168/jds.2024-24994</a>. Oliver MA, Speckhart SL, Edwards JL, Rhoads ML, Ealy AD. 2024. Human recombinant interleukin-6 improves the morphological quality of cryopreserved in vitro produced bovine blastocysts. Theriogenology. 226:173-180. doi: 10.1016/j.theriogenology.2024.06.014.  Ott TL, Tibary A, Waqas M, Geisert R, Giordano JO. 2024. Pregnancy establishment and diagnosis in livestock. Annual Review of Animal Biosciences, 13. <a href="https://doi.org/10.1146/annurev-animal-021022-032214">https://doi.org/10.1146/annurev-animal-021022-032214</a> Plewes MR, Talbott HA, Schott MB, Wood JR, Cupp AS, Davis JS. 2024. Unraveling the role of lipid droplets and perilipin 2 in bovine luteal cells. FASEB J 38:e23710, PMID: 38822676. Przygrokzka E, Binderwala F, Powers R, McFee RM, Cupp AS, Wood JR, Davis JS. 2024. Metabolic control of luteinizing hormone-responsive ovarian steroidogenesis. J. Biol Chem 301:108042 PMID: 39615688, PMCID: PMC11732475. Rial C, Giordano JO. 2024. Combining reproductive outcomes predictors and automated estrus alerts recorded during the voluntary waiting period identified subgroups of cows with different reproductive performance potential. J. Dairy Sci. 107:7299-7316. <a href="https://10.3168/jds.2023-24309">https://10.3168/jds.2023-24309</a> Roach CM, Mayorga EJ, Baumgard LH, Ross JW, Keating AF. 2024. Heat stress alters the ovarian proteome in pre-pubertal gilts. Journal of Animal Science. doi: 10.1093/jas/skae053. PMID: 38605681; PMCID: PMC11025630. Roach CM, Mayorga EJ, Baumgard LH, Ross JW, Keating AF. 2024. Phenotypic, endocrinological, and metabolic effects of zearalenone exposure and additive effect of heat stress in prepubertal female pigs. Journal of Thermal Biology. 119:103742. doi: 10.1016/j.jtherbio.2023.103742.  Roach CM, Mayorga EJ, Baumgard LH, Ross JW, Keating AF. 2024. Zearalenone exposure differentially affects the ovarian proteome in prepubertal gilts during thermal neutral and heat stress conditions. Journal of Animal Science. 102:skae115. doi: 10.1093/jas/skae115. Rishi JK, Timme K, White HE, Kerns KC, Keating AF. 2024. Altered histone abundance as a mode of ovotoxicity during 7,12-dimethylbenz[a]anthracene exposure with additive influence of obesity. Biology of Reproduction. 110(2):419-429. PMID: 37856498. DOI: 10.1093/biolre/ioad140. Rishi JK, Timme K, White HE, Kerns KC, Keating AF. 2024. Trajectory of primordial follicle depletion is accelerated in obese mice in response to 7,12-dimethylbenz[a]anthracene exposure. Biology of Reproduction. 111(2):483-495. doi: 10.1093/biolre/ioae059. Sahoo PK, Krishnamoorthy C, Wood JR, Hanson C, Anderson-Berry A, Mott JL, Natarajan SK. 2024. Palmitate induces integrated stress response and lipoapoptosis in trophoblasts. Cell Death and Disease, 15:31, PMID: 38212315  Schalich KM, Koganti PP, Castillo JM, Reiff OM, Cheong SH and Selvaraj V. 2024. The uterine secretory cycle: recurring physiology of endometrial outputs that setup the uterine luminal microenvironment. Physiological Genomics. 56(1):74-97. <a href="https://doi.org/10.1152/physiolgenomics.00035.2023">https://doi.org/10.1152/physiolgenomics.00035.2023</a> Senn, L. K., K. D. Peterson, J. L. Edwards, R. R. Payton, and D. J. Mathew. Oviduct and endometrial epithelium improve IVP bovine embryo developmental kinetics. Reproduction. 2024. 167 e240008.  doi:10.1530/REP-24-0008. Speckhart SL, Oliver MA, Keane JA, Dias NW, Mercadante VRG, Biase FH, Ealy AD. 2024. Interleukin-6 supplementation improves bovine conceptus elongation and transcriptomic indicators of developmental competence. Biol Reprod. Jul 12;111(1):43-53. doi: 10.1093/biolre/ioae045. Studer JM, Kiefer ZE, Koester LR, Johnson EM, Schmitz-Esser S, Farkas A, Galina Pantoja L, Vonnahme KA, Greiner LL, Keating AF, Baumgard LH, Ross JW. 2024. Evaluation of circulating immune cells, analytes, and inflammatory markers in sows affected with postpartum dysgalactia syndrome. Journal of Animal Science. doi: 10.1093/jas/skae270.  Sun F Ali NN, Londoño-Vásquez D, Simintiras CA, Qiao H, Ortega MS, Agca Y, Takahashi M, Rivera RM, Kelleher AM, Sutovsky P, Patterson AL, Balboula AZ. Increased DNA damage in full-grown oocytes is correlated with diminished autophagy activation. Nature Communications. 2024. 19: 9463. Timme K, González-Alvarez, ME, Keating AF. Pre-pubertal obesity compromises ovarian oxidative stress, DNA repair and chemical biotransformation. Toxicology and Applied Pharmacology. 2024. 489:116981. doi: 10.1016/j.taap.2024.116981. Book Chapters (2024) Bishop CV. Corpus Luteum (Encyclopedia of Reproduction, 3rd Edition). 2024. Elsevier. González-Alvarez ME, Kiefer ZE, Studer JM, Ross JW, Keating AF. 2024. Understanding ovarian function in pigs. Advances in pig breeding and reproduction. In Press. BDS Publishing. J.W. Ross (Ed.). Conference Proceedings (2024) Garcia Guerra A, Motta JCL, Hayden CB, Sala RV, Pereira DC, Absalon-Medina A, Moreno JF, Ross. J. Manejo reproductivo de la donante para la producción exitosa de embriones in vitro. (Translation: Reproductive management of the donor for successful in vitro embryo production). Proceedings of the 9° Simposio Internacional Avances en Reproducción Bovina (9th International Symposium Advances in Bovine Reproduction), August 1-2nd, 2024, Guadalajara, Jalisco, México, Pages 1-17. Garcia Guerra A, Sala RV, Motta JCL, Wiltbank MC. Manejo de receptoras de embriones para una reproduccion exitosa: fertilidad y eficiencia. (Translation: Management of Embryo Recipients for Successful Reproduction: Fertility and Efficiency). Proceedings of the 9° Simposio Internacional Avances en Reproducción Bovina (9th International Symposium Advances in Bovine Reproduction), August 1-2nd, 2024, Guadalajara, Jalisco, México, Pages 119-137. Garcia Guerra A, Rojas Cañadas E, Crist AE, Folchini NP, Mussard ML. Avances en programas de sincronizacion para IATF en ganado de carne: estrategias de presincronización. (Translation: Advances in synchronization programs for fixed time AI in beef cattle: presynchronization strategies). Proceedings of the XXVI Congreso Internacional Anembe de Medicina Bovina (XXVI International Congress of Bovine Medicine Anembe) 2024, April 24 - 26, Córdoba, Spain, Pages 200-212. Garcia Guerra A, Duran B, Motta JCL, Sala RV, Wiltbank MC. Perdidas gestacionales: bases fisiologicas y estrategias para reducirlas. (Translation: Pregnancy loss: physiological bases and mitigating strategies). Proceedings of the XXVI Congreso Internacional Anembe de Medicina Bovina (XXVI International Congress of Bovine Medicine Anembe) 2024, April 24 - 26, Córdoba, Spain, Pages 184-199. Ting A, Weegman B, Bishop CV, Wowk B, Chow L, Pagotan R, Stricklin OE, Fox AK, Harrington SS, Weiss T, Estill C, Fahy G, Zelinski M. (2024). Functional preservation after ovarian tissue and whole ovary vitrification and auto-transplantation in large animal models (vol. 117, pp. 105023). Cryobiology <a href="https://www.sciencedirect.com/science/article/pii/S0011224024001780">https://www.sciencedirect.com/science/article/pii/S0011224024001780</a>. Abstracts (2024) Adeyanju O, González-Alvarez ME, Antwi-Boasiako, C, Polivanov LJ, Shelton J, Nordell NE, Sillman SJ, Laws MJ, Raetzman L, Flaws JA, Desaulniers AT, Keating AF. 2024. Female offspring ovarian chemical metabolism protein abundance is altered by in utero atrazine exposure. Society for Study of Reproduction annual meeting. Alward KJ, Oliver MA, Johnson SE, Ealy AD. In Vitro Embryo Production Can Be Achieved After Delayed Harvest of Bovine Oocytes from Abattoir-derived Ovaries. 2024. Society for the Study of Reproduction Annual Meeting, Dublin Ireland. Antwi-Boasiako C, González-Alvarez ME, Romoser MR, Ross JW, Baumgard LH, Keating AF. 2024.  The effects of heat stress on the abundance of proteins involved in chemical biotransformation in the corpora lutea of post-pubertal gilts. Society for Study of Reproduction annual meeting. Baber T, Newman S, Joyce K, Rhoads ML. 2024. Relationships between circulating glucose concentrations and productivity in heat-stressed dairy cattle. J. Dairy Sci. 107(suppl 1): 320. Bishop CV, RancHER. 2024. Strategies for Preparing Low Parity Heifers and Cows for Their Next Breeding Season, Online. National, Invited.. Bowne S, Ting A, Bishop CV. 2024. Northwest Reproductive Sciences Symposium, "Use of Anti-Mullerian Hormone to Determine Oocyte Aspiration Success in Synchronized Sheep.," Bend, OR. Regional, Accepted. Branco L. Henning J, Rocha, CC. 2024. Rescuing the weight loss caused by toxic calves in calves. CAFNR Research Symposium, Columbia MO. Carothers A , Diaz FJ. 2024. Role of Zinc Insufficiency in Bovine Granulosa Cell Viability. 57th annual meeting of the Society for the Study of Reproduction, July 15-19, 2024, Dublin, Ireland (poster presentation). Crist AE, Motta JCL, Hayden CB, Rykaczewski C, Mussard MM, Garcia-Guerra A. 2024. Characterization of anovular phenotypes in postpartum beef cows. Reproduction Fertility and Development 36(2):230-231. DOI: <a href="https://doi.org/10.1071/RDv36n2Ab154">10.1071/RDv36n2Ab154</a> Davis JS, Przygrodzka E, Plewes MR. 2024. Control of Luteal Function by Luteinizing Hormone: Insights into multi-organelle control of steroidogenesis. The 6th International Conference on Uterine Disorders and Corpus Luteum Function. September 9–12, 2024, Wroclaw, Poland. Ermisch AF, Shao J, Beede K, Schmaltz R, Ramer-Tait A, Wood JR. 2024. Luteinizing hormone stimulation of germ-free and conventionalized female mice reveals microbiome-dependent effect on ovarian steroidogenesis and ovulation. 57th Annual Meeting of the Society for the Study of Reproduction, Dublin, Ireland.  Ermisch AF and Wood JR. 2024. Comparison of N6-methyladenosine modification of mRNA during the bovine and murine oocyte to embryo transition. 50th Annual Meeting of the International Embryo Transfer Society, Denver, CO.   Folchini NP, Crist AE, Motta JCL, Wellert S, Rykaczewski C, Saad M, Carranza Martin AC, Garcia-Guerra A. 2024. Expression of estrus and fertility of different presynchronization strategies for fixed-time artificial insemination in suckled beef cows. Reproduction Fertility and Development 36(2):244-245. DOI: <a href="https://doi.org/10.1071/RDv36n2Ab181">10.1071/RDv36n2Ab181</a> Fox AK, Hammerich MS, Bishop CV. 2024. Northwest Reproductive Sciences Symposium, Vitamin A and D Pre-exposure to Prime Reproductive Success in Breeding Beef Cows. Bend, OR. Regional, Accepted.   Garcia-Guerra A, Motta JCL, Crist AE, Folchini NP, Rykaczewski C, Wellert S. 2024. Optimization of the time of insemination and presynchronization treatment in beef heifers submitted to fixed-time artificial insemination. Reproduction Fertility and Development 36(2):243. DOI: <a href="https://doi.org/10.1071/RDv36n2Ab178">10.1071/RDv36n2Ab178</a> Giordano JO. 2024. Ovarian physiology of previously inseminated cows: Considerations for rebreeding. J. Dairy Sci. 107 (E-Suppl. 1). <a href="https://www.adsa.org/Meetings/Past-Meetings/2024">https://www.adsa.org/Meetings/Past-Meetings/2024</a> González-Alvarez ME, Timme K, Keating AF. 2024. Effect of DMBA exposure on ovarian chemical metabolism, DNA damage sensing and repair, and oxidative stress proteins in prepubertal mice. Society for Study of Reproduction annual meeting. Haffner C, Rishi JK, Keating AF. 2024. Altered abundance of proteins involved in chemical biotransformation in inguinal adipose tissue in lean and obese female mice during exposure to dimethylbenz[a]anthracene. Society for Study of Reproduction annual meeting. Henning J, Branco L, Rocha CC. 2024. Using anogenital distance to predict probability of pregnancy to artificial insemination in beef heifers. CAFNR Research Symposium, Columbia MO. Heredia DC, Tarnonsky F, Lopez-Duarte MC, Venturini M, Ojeda OA., Luchini D., Hansen PJ, Pringle D, DiLorenzo N, Gonella-Diaza AM. 2024. Impact of supplementing rumen-protected methionine during the periconceptional period in the post-weaning period in female calves, Journal of Animal Science, 102 (S1): 69–70, <a href="https://doi.org/10.1093/jas/skae019.081">DOI: 10.1093/jas/skae019.081</a> Hughes CHK, Hellmers AC, daSilva MI, Ott TL, Pate JL. 2024. IFNT-dependent and IFNT-independent changes in the CL of early pregnancy. 57th annual meeting of the Society for the Study of Reproduction, July 15-19, 2024, Dublin, Ireland (poster presentation). Hughes J, Hughes CHK. 2024. Proteomic profiling reveals potential mechanisms regulating prostaglandin F2A responsiveness in bovine corpora lutea. Annual meeting of the American Dairy Science Association, June 16-19, 2024, West Palm Beach, FL (poster presentation). Keane JA, Oliver MA, Pollock AB, Alward KJ, McGehee JF, Kesler ML, Snider AP, Ealy AD. Impacts of OmniGen-AF® Feed Supplementation on Ovum Pick-Up Collections in Beef Cattle. Society for the Study of Reproduction Annual Meeting, Dublin Ireland. Laplacette AL,  Stangaferro ML, Thomas MJ, Giordano JO. 2024. Reproductive performance of lactating dairy cows managed with a targeted reproductive program aimed at increasing AI in estrus after synchronization of ovulation. J. Dairy Sci. 107 (E-Suppl. 1). <a href="https://www.adsa.org/Meetings/Past-Meetings/2024">https://www.adsa.org/Meetings/Past-Meetings/2024</a> Laplacette AL,  Stangaferro ML, Thomas MJ, Giordano JO. 2024. A reproductive management program aimed at maximizing insemination of cows at detected estrus after synchronization of ovulation improved first service outcomes. J. Dairy Sci. 107 (E-Suppl. 1). <a href="https://www.adsa.org/Meetings/Past-Meetings/2024">https://www.adsa.org/Meetings/Past-Meetings/2024</a> Laws MJ, Kramer S, Weis K, Polivanov LJ, Shelton J, Nordell NE, White BR, Sillman SJ, Raetzman L, Keating AF, Flaws JA, Desaulniers AT. 2024. Environmentally Relevant Maternal Atrazine Exposure Leads to Aberrant Endocrine Effects in Piglets. Society for Study of Reproduction annual meeting. Mathew DJ, Peterson KD, Freeman TF, Oliver MA, Kakhniashvili D, Johnson DL, Payton RR, Prado TM. Strickland LG, Edwards JL, Beever JE. 2024. Processes and pathways associated with in vivo derived and in vitro produced male and female bovine conceptus proteins. 57th Annual Society for the Study of Reproduction (SSR), Dublin, Ireland. Abstract and poster presentation. Moorey SE. Role of the preovulatory follicle in oocyte metabolic and developmental competence for pregnancy. 2024. American Society of Animal Science / Canadian Society of Animal Science / Western Section American Society of Animal Science Joint Annual Meeting; Biennial Reproduction Symposium. Calgary, Alberta, Canada. Invited abstract and oral presentation. Motta JCL, Carranza-Martin AC, Rykaczewski C, Folchini NP, Saad M, Hayden C, Sala RV, Bond R, Pereira DC, Ross PJ, Garcia-Guerra A. 2024. Anti-Müllerian hormone: how early can it be used as a biomarker for future in vitro embryo production in Bos taurus cattle? Reproduction Fertility and Development 36(2):250-251. DOI: <a href="https://doi.org/10.1071/RDv36n2Ab191">10.1071/RDv36n2Ab191</a> Nikolaou TA, Rial C, Hussain I, Giordano JO, Erickson D. 2024. ReproPhone: A lateral flow based portable platform for the determination of cattle reproductive status. Biomedical Engineering Society. 2024. Baltimore, MD., October 2024. Oliver MA, Johnson SE, Ealy AD. 2024. Supplementation of Leukemia Inhibitory Factor or Interleukin 6 Modifies the Cellular Lineage Composition within the Embryonic Disc of Extended Cultured Bovine Blastocysts. Society for the Study of Reproduction Annual Meeting, Dublin Ireland.  Pollock AB, Oliver MA, Johnson SE, Ealy AD. 2024. Chemical Inhibition of IL6ST Compromises the In Vitro Production of Bovine Embryos. Society for the Study of Reproduction Annual Meeting, Dublin Ireland.  Reamsnyder T, Sala RV, Pinete-Gonzalez A, Maldonado E, Robles-Morado I, Martinez LC, Absalon-Medina VA, Fricke VC, Juarez-Dorantes RI, Ross PJ, Moreno JF, Motta JCL, Garcia-Guerra A. 2024. “Effect of length of progesterone device administration during a modified CO-Synch for fixed-time embryo transfer in heifers” Reproduction Fertility and Development 36(2):209-210. DOI: <a href="https://doi.org/10.1071/RDv36n2Ab114">10.1071/RDv36n2Ab114</a> Rishi JK, Montes C, Walley JW, Keating AF. 2024. Obesity alters sumoylation-dependent proteomic response in ovaries of mice exposed to 7,12-dimethylbenz[a]anthracene. Society for Study of Reproduction annual meeting. Roa-Vidal P, Markway M, Moraes JGN, Lucy MC, Patterson AL. 2024. Uncovering the spatial gene expression of the bovine uterus: a novel study model for adenomyosis. Poster presented at the Gilbert S. Greenwald Symposium Annual Meeting, Kansas City Kansas. Roa-Vidal P, Medeiros de Almedia J, Wang Z, Patterson AL. 2024. Establishing a bovine in vitro model to investigate the mechanisms of adenomyosis development. Poster presented at the Translational Bioscience PhD Program Symposium, MU School of Medicine, Columbia MO. Rose PA, Bidne KL, McCain AR, Shankar K, Wood JR. 2024. Maternal obesity is associated with global reductions in gene expression including mRNAs that contribute to cell migration and glucocorticoid metabolism in the mid-gestation mouse placenta. 57th Annual Meeting of the Society for the Study of Reproduction, Dublin, Ireland. Saad M, Duran BJ, Rykaczewski C, Kieffer JD, Menchaca A, Garcia-Guerra A. 2024. Effect of P.G. 600 dose on expression of estrus, fertility, and prolificacy in ewes synchronized out-of-season using a short-term progesterone-based protocol. Reproduction Fertility and Development 36(2):245. DOI: <a href="https://doi.org/10.1071/RDv36n2Ab182">10.1071/RDv36n2Ab182</a> Sala RV, Motta JCL, Absalon-Medina VA, Fricke VC, Crist AE, Reamsnyder T, Ross PJ, Moreno JF, Garcia-Guerra A. 2024. Assessment of porcine follicle-stimulating hormone delivery mode before ovum pickup and in vitro embryo production in pregnant heifers. Reproduction Fertility and Development 36(2):250. DOI: <a href="https://doi.org/10.1071/RDv36n2Ab190">10.1071/RDv36n2Ab190</a> Sellmer Ramos I, Caldeira MO, Patterson AL, Lucy MC. 2024. Spatial transcriptomics of the neonatal bovine uterus reveals potential markers of epithelial differentiation driving endometrial adenogenesis. Poster presented at the Society for the Study of Reproduction Annual Meeting, Dublin Ireland. Sitko EM, Giordano JO. 2024. Effect of prioritizing AI in estrus or timed AI and milk yield on cash flow of primiparous cows. J. Dairy Sci. 107 (E-Suppl. 1). <a href="https://www.adsa.org/Meetings/Past-Meetings/2024">https://www.adsa.org/Meetings/Past-Meetings/2024</a> Sitko EM, Laplacette AL, Duhatschek D, Rial C, Perez MM, Tompkins S, Kerwin AL, Domingues RR, Wiltbank MC, Giordano JO. 2024. The&ensp;ovarian function and endocrine phenotypes of lactating dairy cows were&ensp;associated with genomic merit for fertility. J. Dairy Sci. 107 (E-Suppl. 1). <a href="https://www.adsa.org/Meetings/Past-Meetings/2024">https://www.adsa.org/Meetings/Past-Meetings/2024</a> Timme K, González-Alvarez ME, Keating AF. 2024. Both obesity and DMBA exposure alter ovarian follicle number, ZP3 and PPP2CA in prepubertal females. Society for Study of Reproduction annual meeting.  Venturini M, Heredia DC, Lopez MC, Joice K, Ruiz M, Dubeux JCB,  Dilorenzo N, Gonella-Diaza AM. 2024. Plasma metabolomics and feed behavior patterns in feed efficient or inefficient beef bulls. Journal of Animal Science, 102 (S1): 1–2, <a href="https://doi.org/10.1093/jas/skae019.001">DOI: 10.1093/jas/skae019.001</a> Weiss T, Ting A, Bishop CV. 2024. Northwest Reproductive Sciences Symposium, Optimization of Immunohistochemistry Techniques for the Use of Polyclonal Rabbit Anti-Human von Willebrand Factor to Detect Ovarian Vascularization in Ovine Cryopreserved Ovaries, Bend, OR. Regional, Accepted. Wiltbank MC, Andrade JP, Gomez-Leon V, Garcia-Guerra A, Monteiro PLJ, Domingues RR, Fricke PM, Sartori R. 2024. The dichotomy between pregnancy survival and loss: Challenging post-conceived notions. Journal of Dairy Science 107(Supp 1):170.

Impact Statements

1) Obtaining a greater basic understanding of the complex, molecular regulation of bovine ovarian blood flow and cellular function during the estrous cycle to minimize infertility and pregnancy losses.
2) Understanding the impacts of environmental exposures on female reproductive function will enable us to develop schemes to reduce the adverse effects of heat stress, and endogenous and exogenous chemical exposures in cattle and other mammals.
3) Assessing mouse and pig models to understand basic biological impacts of these exposures as well as to develop targets to mitigate negative effects. The impacts of these findings are that the scientific, clinical and production animal communities benefit from the advanced knowledge provided.
4) Insight gained from research describing how zinc, an essential micronutrient, influences bovine mural granulosa cell proliferation will advance the understanding of the mechanisms that govern the maintenance of pregnancy, and this ultimately will allow for the development of strategies to reduce pregnancy loss and, thus, increasing reproductive efficiency.
5) Advancing our mechanistic understanding of bovine conceptus elongation will lead to new strategies for reducing early pregnancy losses, thus enhancing reproductive efficiency and economic viability in cattle production systems.
6) Optimizing ovarian function, embryo quality, and uterine receptivity to pregnancy are key areas that must be addressed to improve reproductive success in cattle. Insight gained from this research will provide important information to advance the understanding of the mechanisms that govern the ovarian follicle’s contribution to oocyte competence for embryo development and the uterine-conceptus relationships required for pregnancy establishment and maintenance. Ultimately, by characterizing physiologically normal follicular and uterine environments, and determining deficiencies during assisted reproduction, strategies to overcome limitations of assisted reproductive techniques can be developed.
7) Identifying the critical physiological responses to heat stress and determining their overall impact on reproduction, health and well-being will equip researchers and producers with the information needed to effectively improve the management of heat-stressed cattle.
8) Fundamental knowledge of the mechanisms regulating the health of the follicle, and ultimately improving fertility of dairy cows, will impact the >$2.2B dairy industry (annually) in the states of Vermont and Rhode Island.
9) Targeted reproductive management (TRM) of dairy cattle will increase dairy farm profitability and sustainability through tailored reproductive management schemes based on data for predictors of variability in cow biology and expected performance.
10) Understanding the molecular and cellular basis regulating ovarian and uterine physiology in dairy cattle will provide new opportunities for developing targeted strategies to improve pregnancy outcomes in dairy cows.
11) Providing modest increases in the availability of vitamins A, D and E can be a relatively inexpensive method to improve fertility, and this scheme may become increasingly relevant in future years due to climate impacts on production.
12) Providing new insights into the stability, translation, and degradation of oocyte mRNAs is regulated during oogenesis, embryogenesis, and the first stages of placentation will provide new insights into how alterations in mRNA metabolism influences oocyte quality.
13) Highly steroidogenic granulosa cells are essential for the growth and development of the oocyte and the eventual establishment of pregnancy. Understanding the mechanisms that control granulosa cell function has important implications that will ultimately lead to improving fertility by developing new strategies to enhance reproductive outcomes and assisted reproduction technologies.
Student training is another important impact of this project. Members of the NE227 group (NY, IA, MS, OH, OR, PA, KY, MA, NE, TN, VT, WI, VA) developed a course on Contemporary Topics in Reproductive Biology to improve student understanding of the breadth of reproductive physiology including topics outside their primary area of research. Greater than 50 students have enrolled in the fall 2024 offering. This accounted for a majority of the MS and PhD students across the experiment stations in this multistate project. Numerous undergraduate students with an interest in reproductive physiology were also introduced to investigative research. These activities represent an important contribution of the project to the education of the next generation of scientists, consultants, and other workers in animal agriculture industries.