Progress 10/01/09 to 09/30/14
Outputs
Target Audience: LSU AgCenter-related departments and units who utilize the Reproductive Biology Center (RBC) for support of their research, teaching, and extension needs. The RBC provides land, facilities, equipment, and personnel to assist with these activities. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?
Nothing Reported
How have the results been disseminated to communities of interest?
Nothing Reported
What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
Services were provided to the LSU AgCenter School of Animal Sciences, the Dean Lee Research Station, and the Department of Entomology in support of reproductive research and teaching activities.
Publications
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Progress 01/01/13 to 09/30/13
Outputs
Target Audience: Target audience groups include all LSU AgCenter-related departments and units who utilize the Reproductive Biology Center (RBC) for support of their research, teaching, and extension needs. The RBCprovides land, facilities, equipment, and personnel to assist with these activities. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?
Nothing Reported
How have the results been disseminated to communities of interest?
Nothing Reported
What do you plan to do during the next reporting period to accomplish the goals? Continue to provide support of animal research to AgCenter departments and units.
Impacts
What was accomplished under these goals?
Services were provided to the LSU AgCenter School of Animal Sciences, the Dean Lee Research Station,and the Department of Entomology in support of reproductive research and teaching activities.
Publications
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Progress 01/01/12 to 12/31/12
Outputs
OUTPUTS: Research from this project resulted in one book chapter, 2 publications in a peer reviewed journal and 4 peer reviewed abstracts presented at international meetings. PARTICIPANTS: G. Gentry (PI), LSU AgCenter. TARGET AUDIENCES: Scientists involved in reproductive biology of domestic and wild animals. PROJECT MODIFICATIONS: Not relevant to this project.
Impacts
Assisted reproductive technologies are a tool that can be utilized to increase the reproductive efficiency and genetic merit of domestic livestock. For cattle fixed-timed artificial insemination (FTAI) has not been a technique incorporated into the management schemes of beef producers due to variability of resulting pregnancy rates. Studies from our laboratory showed that following FTAI, pregnancy rate was not affected by administration of follicle stimulating hormone (FSH) during the synchronization period and overall pregnancy rates was 50%. Similarly, based on response to synchronization, there was no difference in the percentage of females responding to the synchronization protocol and no difference in the pregnancy rate of those responding females for the FSH treated and non-treated control. Therefore we concluded administration of FSH ten days prior to FTAI in CO-Synch+CIDR synchronized crossbred beef cows and heifers may not be associated with increased pregnancy rates. In another study, we collected pre-implantation stage equine embryos between day 6 and 8 following artificial insemination from light horse mares. Embryos averaged a diameter of 526 microns and ranged from 243 to 1072 microns. Embryos were stratified by diameter and subjected to either a single puncture or double puncture of the capsule to allow entry of vitrification medium into the embryo proper. Following treatment, embryos were equilibrated in vitrification solutions following standard laboratory protocol. Embryos were then incubated in culture medium for 72 h and re-expansion recorded. Thirty percent of the embryos hatched in the double puncture groups and none hatched in the single puncture group. After the 72 h culture, 70% of the single puncture embryos and 50% of the double puncture group were considered still viable. Statistical comparisons across treatments are now underway. We have investigated methods for the cryopreservation of semen and embryos of domestic species. Successful cryopreservation of gametes is a major component of any advanced breeding program for domestic animals. Cryopreserved gametes aid in the application of selective breeding programs, international movement of genetic material and banking of genetic material for future use. Studies conducted as part of this project provide new insight into how various factors influence the success or failure of these processes. Somatic Cell Nuclear Transfer represents a powerful tool for the production of genetically modified livestock animals to meet the needs of increased food production while minimizing the impact on the environment. Induced pluripotency is an important new technology for creating undifferentiated cell lines which can be used for nuclear transfer. In order for these technologies to be applicable to domestic animals, efficiency must be increased and the induction of pluripotency must be accomplished without altering the genome of the somatic cells. Results from this project provide initial progress towards achieving induced pluripotency in bovine somatic cells without genetic alteration.
Publications
- Stout, M.A., J.R. Sanez, J. Chenevert, G.T. Gentry, K.R. Bondioli and R.A. Godke. 2012. Cryopreserved ejaculated and epididymal sperm collected from the same Holstein bulls utilized for in vitro fertilization. Proc. Gulf Coast Conservation Biology Symposium 8:5.
- Steinmiller, J.L., K. Bondioli, R. Godke and G.T. Gentry. 2012. In vitro fertilization media and composition agent effects on epididymal white-tailed deer spermatozoa. Proc. Gulf Coast Conservation Biology Symposium 8:11.
- Scott, B.R., D.B. Carwell, R.A. Hill, K.R. Bondioli, R.A. Godke and G.T. Gentry. 2012. Evaluation of capsule permeability in the equine blastocyst. J. Equine Vet Sci. 32:795-798.
- Gentry, G.T., J. Lambe, W. Forbes, B. Olcott, D. Sanders, K. Bondioli and R.A.Godke. 2012. The effect of eCG on pregnancy rates of white-tailed deer following fixed-timed artificial insemination Theriogenology 77: 1894-1899.
- Giraldo, A. M., S. Ball, and K. R. Bondioli. 2012. Production of transgenic and knockout pigs by somatic cell nuclear transfer. Methods Mol Biol 885: 105-123.
- Carwell, D. B., B. R. Scott, J. Len, H. Blackburn, K. R. Bondioli, G. T. Gentry, and R. A. Godke. 2012. Effect of storage duration on post-thaw parameters of bull semen. Reproduction, Fertility and Development 25: 177-177.
- Adams, T. L., S. E. Farmer, R. A. Godke, and K. R. Bondioli. 2012. Stimulation of endogenous oct4 in bovine fetal fibroblasts with synthetic mRNA transfection. Reproduction, Fertility and Development 25: 296-297.
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Progress 01/01/11 to 12/31/11
Outputs
OUTPUTS: Research from this project resulted in 1 publication in a peer-reviewed journal and 3 peer-reviewed abstracts presented at international meetings. A trial with aminocyclopyrcholr alone and in combination led to the inclusion of sugarberry (Celtis laevigata) in the new herbicide Streamline label. PARTICIPANTS: Gentry, G.T. (PI), LSU AgCenter. TARGET AUDIENCES: Nothing significant to report during this reporting period. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
Studies from our laboratory showed that bovine epididymal sperm displayed lower levels of cryo-acrosome reaction but were similar by level of cryo-capacitation compared with ejaculated bovine sperm. In addition, epididymal and ejaculated sperm differed in the need for a capacitating agent such as heparin when used in vitro. We did not detect a difference in Oct-4 and Nanog transcript levels between untreated bovine fetal fibroblasts (BFF) cells and BFF cells exposed to adipose derived adult stem (ADAS) cell extracts. Comparison between untreated BFF cells and BFF cells treated with ADAS cell extracts revealed a higher Sox-2 expression in the extract-treated cells. We also showed that there was no difference in the number of follicles aspirated in cattle administered either 3 injections of FSH in saline every 12 h, 60 h following dominant follicle removal (DFR) or a single injection of FSH in saline coincident with FSH and PVP 60 h following DFR and undergoing transvaginal ultrasound guided aspiration. We evaluated the association of cow temperament and artificial insemination (AI) pregnancy rates in crossbred beef females. We found that cortisol concentrations are correlated with exit velocity and that females becoming pregnant to AI had lower exit velocity than females not conceiving to AI. We showed hybridization of red deer and white-tailed deer through laparoscopic insemination of red deer hinds with white-tailed semen and white-tailed does with red deer semen needs to be evaluated further, and in year one those inseminations resulted in no pregnancies. Also, year six of white-tailed deer artificial insemination was initiated; and 40 does were inseminated for fixed-timed AI, where timing of insemination was adjusted based on prior years results. Pregnancy rates are not yet available. Data from multiple brush and herbaceous weed trials indicated that the new herbicide aminocylopyrchlor was very effective in controlling most herbaceous species and many brush weed species at active ingredient rates 90% less than competitive herbicides, thereby greatly reducing the potential amount of herbicide released into the environment.
Publications
- Addison MK, Coley LW, Gentry GT, Godke RA, Bondioli KR. Epigenetic modification with zebularine and valproic acid and expression of pluripotency genes in bovine adipose stem cells. Reprod Fertil Dev 2011;24:216-17.
- Coley LW, Addison MK, Gentry GT, Godke RA, Bondioli KR. Reprogramming of bovine somatic cells with cellular extracts from adipose-derived adult stem cells and human embryonic stem cells.Reprod Fertil Dev 2011;24:216-16.
- Stout MA, Saenz JR, Chenevert JF, Gentry GT, Bondioli KB, Godke RA. Cryo-acrosome reaction/capacitation of ejaculated and epididmyl bovine sperm and their in vitro fertilization rates. Reprod Fertil Dev 2011;24:189-89.
- Stamatkin CW, Roussev RG, Stout M, Coulam CB, Triche E, Godke RA, Barnea ER. Preimplantation factor negates embryo toxicity and promotes embryo development in culture. Reproductive BioMedicine Online 2011;23:517-24.
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Progress 01/01/10 to 12/31/10
Outputs
OUTPUTS: Research from this project resulted in 4 publications in peer reviewed scientific journals, 3 book chapters, 3 thesis and 10 abstracts presented at international meetings. PARTICIPANTS: G. T. Gentry, R. A. Godke, K. R. Bondioli, L. D. Foil, J. A. Pitchford and a number of graduate students, plus collaborators from Audubon Center for Research of Endangered Species. TARGET AUDIENCES: Livestock producers and the scientific community. PROJECT MODIFICATIONS: Nothing significant to report during this reporting period.
Impacts
Studies in our laboratory showed that equine embryos incubated in tritiated glycerol at two concentrations for 15 min exhibited no difference in glycerol uptake (1.4M- 2.6% vs. 3.4M-1.2%). Embryos with a diameter less than 400 um in the 1.4M treatment took up significantly more labeled glycerol than embryos with a diameter greater than 400 um (P<0.05). Because the capsule may not be fully developed in these smaller embryos, these data suggest that the capsule may be a cryoprotectant permeability barrier. Our laboratory also showed that the inclusion of serum to culture medium for in vitro produced embryos can change the expression of key genes related to placental function in some embryos.These changes in gene expression were seen at the blastocyst stage (7 days) and in elongated embryos (14 days). We have also reported that frozen-thawed semen can be refrozen and upon subsequent thawing result in viability suitable for use in artificial insemination. This indicates that very valuable semen thawed inadvertently does not necessarily have to be discarded but can be refrozen and maintain motility. We have also shown that the addition of a cloth target to an alsynite trap could increase the efficacy of the fly trap by approximately three times. Studies indicate that 3,000 to 4,000 flies per day can be killed using a treated target that is randomly placed in pastures on cattle farms during stable fly season. We also showed that the number of flies landing on targets was nearly five-fold higher when the targets were placed close to cattle. We found that treated targets that are placed around cattle for stable fly control should be placed at ground level. We found a significant increase in flies captured ranging from 3.3 to 2.6 fold when cattle were present on these traps.
Publications
- Foil, L.D., F.D. Guerrero, and K.G. Bendele. 2010. Detection of target site resistance to pyrethroids and organophosphates in the horn fly using multiplex PCR. J Med Ent. 47: 855-861.
- Sansinena, M.J., J. W. Lynn, K. R. Bondioli, R. S. Denniston and R. A. Godke. 2010. Ooplasm transfer and interspecies somatic cell nuclear transfer: Heteroplasmy, pattern of mitochondrial migration and effect on embryo development. Zygote Aug 25:1-10 (Epub ahead of print).
- Giraldo, A. M., D. Hylan, K. R. Bondioli, and R. A. Godke. 2010. Distribution of sexes within the left and right uterine horns of cattle, Theriogenology 73:496-500.
- Owiny, O. D., D.M. Barry and R. A. Godke. 2010. Effects of repeated freeze-thawing of bovine epididymal sperm on in vitro embryo development. Afr. J. Anim. & Biomed. Sciences 5(1):56-64.
- Scott, B. R., D. B. Carwell, R. A. Hill, K. R. Bondioli, R. A. Godke, and G. T. Gentry. 2010. Serum progesterone concentration and embryo diameter on day 7 post-ovulation in mares. Reproduction, Fertility and Development 23: 163-164.
- Carwell, D. B., B. R. Scott, G. T. Gentry, K. R. Bondioli, and R. A. Godke. 2010. Refreezing post-thawed goat semen. Reproduction, Fertility and Development 23: 140-141.
- Addison, M. K., G. T. Gentry, R. A. Godke, and K. R. Bondioli. 2010. Inducing pluripotency in bovine somatic stem cells. Reproduction, Fertility and Development 23: 242-242.
- Angulo, J., G. T. Gentry, R. A. Godke, and K. R. Bondioli. 2010. Effect of serum during culture on day 14 elongated bovine embryos. Reproduction, Fertility and Development 23: 191-191.
- Carwell, D.B., J.A. Pitchford, G.T. Gentry, Jr., H. Blackburn, K.R. Bondioli, and R.A. Godke. 2010. Beef cattle pregnancy rates following insemination with aged frozen angus semen. Reprod. Fertil. Dev. 22:167-168.
- Chiasson, M.K., J.A. Carter, K.R. Bondioli, R.A. Godke and G.T. Gentry. 2010. Laser-assisted zona pellucida hatching in frozen-thawed bovine embryos. Reprod. Fertil. Dev. 22:244.
- Coley, L.W., R.A. Godke, and K.R. Bondioli. 2010. Expression of Pluripotency Associated Genes in Bovine Fetal Fibroblast Cells and Adipose Derived Stem Cells. Reprod. Fertil. Dev. 22:275.
- Picou, A., J.A. Wilson, B. Dresser, G.T. Gentry, R. A. Godke and K.R. Bondioli. 2010. Characterization of bovine adult derived adipose stem cells for use in nuclear transfer. Reprod. Fertil. Dev. 22:353.
- Wilson, J.A., R.A. Godke, and K.R. Bondioli. 2010. Determining gene copy number in transfected caprine fibroblast cells. Reprod. Fertil. Dev. 22:375.
- Adipose Tissue Derived Somatic Stem Cells. In: Gimble, J.M. and Bunnell, B.A. (eds.). Adipose-Derived Stem Cells: Methods and Protocols vol 702. Humana Press. 2010 pp. 77-86.
- Youngs, C.R. and R.A. Godke. 2010. Cattle embryo transfer. In (DR Heldman, MB Wheeler, DG Hoover, Eds.) Encyclopedia of Biotechnology in Agriculture and Food DOI: 10.1081/E-EBAF-120042308.
- Youngs CR, SP Leibo, RA Godke. 2010. Embryo cryopreservation in domestic mammalian livestock species. CAB Reviews: Perspectives in Agriculture, Veterinary Science, Nutrition and Natural Resources Vol. 5, No. 60, 11 pages. doi:10.1079/PAVSNNR20105060.
- Angulo, J. 2010. Effects of Serum Addition to Culture Medium on Gene Expression in Day-7 and Day-14 Bovine Embryos. MS Thesis Louisiana State University. Baton Rouge, LA.
- Chiasson, M. 2010. Laser-Assisted Zona Pellucida Hatching of Frozen-Thawed In Vivo-Produced Bovine Embryos. MS Thesis Louisiana State University. Baton Rouge, LA.
- Wooley, D. 2010. Prepartum Maternal Cortisol Concentration on Cortisol and Immunoglobulin G Concentration in Neonatal Dairy Calves. MS Thesis Louisiana State University. Baton Rouge, LA.
- Williams, K.J., K.R. Bondioli and R.A. Godke. 2010. Karyotypic analysis and epigenetic modifications of cultured porcine adipose tissue-derived stem cells. Reprod. Fertil. Dev. 22:356.
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