USE OF EMBRYO TRANSFER TO IMPROVE FERTILITY OF HEAT-STRESSED DAIRY COWS

• Sponsoring Institution: National Institute of Food and Agriculture
• Project Status: COMPLETE
• Funding Source: SPECIAL GRANT
• Reporting Frequency: Annual
• Accession No.: 0208269
• Grant No.: 2006-34135-17639
• Proposal No.: 2006-04902
• Project Start Date: Sep 1, 2006
• Project End Date: Aug 31, 2007
• Grant Year: 2006
• Program Code: [AH]- (N/A)

Recipient Organization
UNIVERSITY OF FLORIDA
G022 MCCARTY HALL
GAINESVILLE,FL 32611

Performing Department
ANIMAL SCIENCES

Non Technical Summary
Fertility is low in heat-stressed cows because of the effects of heat stress on early embryonic development. This project seeks to overcome embryonic death caused by heat stress by using embryo transfer.

Animal Health Component

67%

Research Effort Categories

Basic

33%

Applied

67%

Developmental

(N/A)

Classification

Knowledge Area (KA)	Subject of Investigation (SOI)	Field of Science (FOS)	Percent
301	3410	1020	15%
301	3410	1050	35%
306	3410	1020	15%
306	3410	1050	35%

Knowledge Area
306 - Environmental Stress in Animals; 301 - Reproductive Performance of Animals;

Subject Of Investigation
3410 - Dairy cattle, live animal;

Field Of Science
1020 - Physiology; 1050 - Developmental biology;

Keywords

dairy cows

heat stress

fertility

embryo transfer

Goals / Objectives
Objective 1: Conduct an experiment to verify previous observations that embryos cultured in the presence of insulin-like growth factor-1 (IGF-1) have higher pregnancy rates following transfer into heat-stressed recipients and to test whether a similar beneficial effect of IGF-1 is observed when embryos are transferred into recipients not subjected to heat stress. Objective 2: Test whether treatment of recipient cows with GnRH after embryo transfer increases pregnancy rate in heat-stressed and non-stressed recipients. Objective 3: Evaluate the role of glucose toxicity in causing a skewed sex ratio in IVP embryos by testing whether reduction of glucose concentration in culture medium increases the proportion of transferrable embryos that are female. Objective 4: Determine whether addition of hyaluronic acid, a component of reproductive tract fluids, to embryo culture medium improves survival of IVP embryos to freezing.

Project Methods
For Objective 1, Experiments are planned to determine 1) whether the beneficial effects of IGF-1 on embryonic survival to stress is also observed when evaluating the stress of vitrification, 2) evaluating embryonic gene expression and morphology to determine cellular or molecular changes in the embryo that could explain increased embryonic survival during heat stress and 3) the role for anti-apoptotic actions of IGF-1 in protecting embryos from heat shock and 4) testing whether IGF-1 improves embryonic survival by increasing embryonic interferon-tau secretion. Objective 2 has been completed and no additional work is planned. Experiments for Objective 3, to evaluate the role of glucose toxicity in causing a skewed sex ratio in IVP embryos, is ongoing and will involve culture of embryos in different glucose concentrations and evaluation of sex ratio by PCR. An experiment for Objective 4 will test whether addition of IGF-1 to culture medium improves survival of embryos to freezing.

Progress 09/01/06 to 08/31/07

Outputs
OUTPUTS: We have shown that embryo transfer with insulin-like growth factor-1 (IGF-1) treated embryos can result in pregnancy rates in heat stressed cows that are equivalent to those achieved using AI in cool weather. We have also established that IGF-1 protects embryos from direct effects of elevated temperature on development and delineated the role of apoptosis and signaling pathways in this effect. We have tested effects of bovine somatotropin and gonadotropin releasing hormone (GnRH) on fertility of heat-stressed cows and, for GnRH, embryo transfer recipients, and shown these are without effect. We have also shown that acceptable pregnancy rates can be achieved using embryo transfer using embryos produced by fertilization with sexed semen. We have characterized some of the cellular changes caused by heat shock that disrupt embryonic development including elucidating developmental changes in regulation of heat-shock induced apoptosis. This information has been transmitted to producers via extension meetings (national and state), publications in Hoard's Dairyman and elsewhere, a CD available to dairymen, and through a website called the Dairy Reproduction Cookbook. We have also made the information available to veterinarians at veterinary and scientific meetings and via review articles in scientific publications. PARTICIPANTS: The following students received degrees as a result of work for this project: Carlos Moises Franco, M.S., 2005, Animal Sciences. Thesis: Strategies to enhance fertility in dairy cattle during summer including use of cryopreservation of in vitro produced embryos. Present position: Owner, Embogen, San Carlos, Bolivia. Dean Jousan, Ph.D., 2006, Animal Sciences (Animal Molecular and Cell Biology), Dissertation: Insulin-like Growth Factor-I and Apoptosis as Determinants of Preimplantation Bovine Embryonic Development. Present position: Assistant Professor, Mississippi State University. Jeremy Block, Ph.D., Animal Molecular and Cell Biology, 2007. Ph.D. Dissertation, Effect of insulin-like growth factor-1 on development and post transfer survival of bovine embryos produced in-vitro. Current position: Postdoctoral Research Associate, University of Florida; co-owner, Embogen, Inc. Luis Augusto de Castro e Paula, Ph.D., Animal Molecular and Cell Biology, 2007. Dissertation title: Role of oxygen and sphingomyelin metabolism in actions of heat shock on the oocyte and embryo. Current position: Research Scientist, EMBRAPA, Campo Grande, Brazil. TARGET AUDIENCES: dairy farmers, dairy veterinarians, reproductive biology and animal science researchers

Impacts
We have been successful in developing procedures for improving pregnancy rate in heat-stressed dairy cows through the use of embryo transfer. Pregnancy rates achieved in heat stressed cows using embryo transfer are equivalent to those achieved using AI in cool weather. This result represents the first solution to the problem of heat-stress induced infertility in dairy cattle. As a direct result of our research, embryo transfer is now being used in the field to improve fertility in heat-stressed cows and two commercial companies provide this service (Transova and Embogen).

Publications

• Brad, A.M., and Hansen, P.J. (2007) The block to apoptosis in bovine two-cell embryos involves inhibition of caspase-9 activation and caspase-mediated DNA damage. Reproduction 134, 789-797.
• de Castro e Paula, L.A. and Hansen, P.J. (2007) Interactions between oxygen concentration and glucose concentration that modulate actions of heat shock on bovine oocytes during in vitro maturation. Theriogenology 68, 763-770.
• Hansen, P.J. (2007) To be or not to be - determinants of embryonic survival following heat shock. Theriogenology, 68S, S40-S48.
• Hansen, P.J. (2007) Exploitation of genetic and physiological determinants of embryonic resistance to elevated temperature to improve embryonic survival in dairy cattle during heat stress. Theriogenology, 68S, S242-S249
• Block, J., Wrenzycki, C., Niemann, H., Herrmann, D., and Hansen, P.J. (2008) Effects of insulin-like growth factor-1 on cellular and molecular characteristics of bovine blastocysts produced in vitro. Mol. Reprod. Dev. 75, 895-903.
• Jousan, F.D., Oliveira, L.J., and Hansen, P.J. (2008) Short-term culture of bovine preimplantation embryos with insulin-like growth factor-I prevents heat shock-induced apoptosis through activation of the phosphatidyl 3-kinase/Akt pathway. Mol. Reprod. Dev. 75, 681-688.
• Loureiro, B., Brad, A.M., and Hansen, P.J. (2007) Heat shock and tumor necrosis factor-α induce apoptosis in bovine preimplantation embryos through a caspase-9 dependent mechanism Reproduction 133, 1129-1137.
• Block, J., and Hansen, P.J. (2007) Interaction between season and culture with insulin-like growth factor-1 on survival of in-vitro produced embryos following transfer to lactating dairy cows. Theriogenology 67, 1518-1529.
• Block, J., Fischer-Brown, A.E., Rodina, T.M., Ealy, A.E., and Hansen, P.J. (2007) The effect of in vitro treatment of bovine embryos with IGF-1 on subsequent development in utero to Day 14 of gestation. Theriogenology, 68, 153-161.
• Niasari-Naslaji, A., Hansen, P.J., and Thatcher, W.W. (2007) Successful cryopreservation of in vitro derived bovine blastocysts in microcapillary pipette tips. Iranian J. Vet. Res. 8, 1-7.
• Jousan, F.D., de Castro e Paula, L.A., Block, J., and Hansen, P.J.(2007) Fertility of lactating dairy cows administered recombinant bovine somatotropin during heat stress. J. Dairy Sci. 90, 341-350.
• Jousan, F.D. and Hansen, P.J. (2007) Insulin-like growth factor-I promotes resistance of bovine preimplantation embryos to heat shock through actions independent of its anti-apoptotic actions requiring PI3K signaling. Mol. Reprod. Dev. 74, 189-196.
• Franco, M., Thompson, P.M., Brad, A.M., and Hansen, P.J. (2006) Effectiveness of administration of gonadotropin-releasing hormone at day 11, 14 or 15 after anticipated ovulation for increasing fertility of lactating dairy cows and non-lactating heifers. Theriogenology, 66, 945-954.
• Franco, M., Block, J., Jousan, F.D., de Castro e Paula, L.A., Brad, A., Franco, J.M., Grisel, F., Monson, R.L., Rutledge, J.J., and Hansen, P.J. (2006) Effect of transfer of one or two embryos and administration of gonadotropin releasing hormone on pregnancy rates of heat-stressed dairy cattle . Theriogenology, 66, 224-233.