Progress 08/01/06 to 08/01/12
Outputs
OUTPUTS: During the course of this project we have conducted a number of studies to address specific issues related to animal reproduction. We demonstrated that hens could be better returned to the laying flock using an oral progesterone analog than starving them and thereby keeping them out of production for weeks. We have demonstrated that stimulating the growth hormone axis can help fetal lambs come closer to their genetic potential and continue to seek support to test that in a reduced nutrient environment. And finally, we have demonstrated that dietary manipulation of progesterone catabolism is possible and could contribute to improving the fertility of dairy cattle. PARTICIPANTS: Not relevant to this project. TARGET AUDIENCES: Not relevant to this project. PROJECT MODIFICATIONS: Not relevant to this project.
Impacts
The work conducted on this project should lead to changes in the layer hen industry, dairy feeding and the managment of animals on limited resource range/pasture.
Publications
- No publications reported this period
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Progress 01/01/11 to 12/31/11
Outputs
OUTPUTS: We have continued to conduct and analyze experiments desribed in the proposal, including culturing primary hepatic cells with various CYP450 and other enzyme inhibitors to begin to assess the relative contribution of different enzyme systems to the catabolism of progesterone. In addition we have gone back and added enzymatic analysis of previously collected tissues to add an understanding of the contribution and effect of treatment on aldoketo reductase family enzymes that also can catabolize progesterone. In addition we are part of a multistate project on swine reproductive physiology. During this year we hosted a symposium for swine producers and industry professionals. In addition, our group has been collecting data in collaboration with an outside group at NDSU to understand how exercise during gestation may alter blood flow to the uterus or placenta and how that alteration may impact nutrient transport by the placenta. A host of laboratory analyses have been conducted and are awaiting analyses. PARTICIPANTS: Matthew E. Wilson, PI Caleb O. Lemley, graduate student Tiffany A. Wilmoth, graduate student TARGET AUDIENCES: Not relevant to this project. PROJECT MODIFICATIONS: Not relevant to this project.
Impacts
We have clearly demonstrated that providing sufficient nutrition to maintain milk production, while simply altering the feedstuffs utilized to stimulate insulin secretion, leads to a dramatic decrease in the activity of the enzymes responsible for progesterone, an increase in progesterone half life and a decrease in the metabolic clearance rate of progesterone. At the same time this hyperinsulinemic diet does not alter feed intake, milk yield or blood flow to the liver. The data indicate that CYP2C is the primary cytochrome responsible for progesterone catabolism in the lactating dairy cow, followed by CYP3A and aldoketo reductase. On the swine project we have analyzed a number of samples, but still need to collect data about the relative vascularity of the placental/endometrial interface so that those data can be submitted for analysis
Publications
- No publications reported this period
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Progress 01/01/10 to 12/31/10
Outputs
OUTPUTS: We have continued to conduct and analyze experiments desribed in the proposal, including culturing primary hepatic cells with various CYP450 and other enzyme inhibitors to begin to assess the relative contribution of different enzyme systems to the catabolism of progesterone. In addition we have gone back and added enzymatic analysis of previously collected tissues to add an understanding of the contribution and effect of treatment on aldoketo reductase family enzymes that also can catabolize progesterone. In addition we are part of a multistate project on swine reproductive physiology. During this year we hosted a symposium for swine producers and industry professionals. In addition, our group has been collecting data in collaboration with an outside group at NDSU to understand how exercise during gestation may alter blood flow to the uterus or placenta and how that alteration may impact nutrient transport by the placenta. A host of laboratory analyses have been conducted and are awaiting analyses. PARTICIPANTS: Matthew E. Wilson, PI Caleb O. Lemley, graduate student Tiffany A. Wilmoth, graduate student TARGET AUDIENCES: Not relevant to this project. PROJECT MODIFICATIONS: Not relevant to this project.
Impacts
We have clearly demonstrated that providing sufficient nutrition to maintain milk production, while simply altering the feedstuffs utilized to stimulate insulin secretion, leads to a dramatic decrease in the activity of the enzymes responsible for progesterone, an increase in progesterone half life and a decrease in the metabolic clearance rate of progesterone. At the same time this hyperinsulinemic diet does not alter feed intake, milk yield or blood flow to the liver. The data indicate that CYP2C is the primary cytochrome responsible for progesterone catabolism in the lactating dairy cow, followed by CYP3A and aldoketo reductase. On the swine project we have analyzed a number of samples, but still need to collect data about the relative vascularity of the placental/endometrial interface so that those data can be submitted for analysis.
Publications
- Lemley, C. O. and M. E. Wilson. 2010. Effect of cytochrome P450 and aldo-keto reductase inhibitors on progesterone decay in primary bovine hepatic cell cultures. Journal of Dairy Science 93:4613-4624.
- Lemley, C. O., K. A. Vonnahme, L. R. Tager, K. M. Krause and M. E. Wilson. 2010. Insulin secretion, cytochrome P450 activity, liver blood flow and progesterone clearance in dairy cows fed a high cornstarch versus a high fiber diet. Journal of Endocrinology 205:233-241.
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Progress 01/01/09 to 12/31/09
Outputs
OUTPUTS: During the reporting period we have continued to analyze the data collected from gilts at day 70, 90 or 110 of gestation during the Fall and Winter of 2008. This includes determining fetal and maternal concentrations of progesterone for comparison to samples previously collected from sheep. We have also continued to assay maternal uterine atrerial and venous blood and fetal umbilical vein and unbilical artery blood for the twenty amino acids responsible for protein synthesis in an attempt to further elucidate the relationship between variation in apparent placental function and apparent nutrient transport. We also have initiated a collaboration with colleagues at North Dakota State University to combine and extend our efforts to understand how the gestation environment that a lamb is exposed to impacts both fetal and postnatal growth. We analyzed IGF-I concentrations in weekly samples collected from pregnant ewes fed above, at or below their requirements during the end of gestation and lactation. We are currently in the process of designing experiments to combine our two experimental paradigms and try to increase our understanding of factors regulating the gestational environment. Finally, we have continued to analyze samples collected from dairy cows fed diets designed to result in divergent insulin secretion, first by increasing the number of enzyme systems we have analyzed in samples collected during the previous year as well as conducting in vitro experiments to better understand the relative contribution of a number of enzyme systems to progesterone inactiviation. PARTICIPANTS: Wilson, Matthew E. Krause, K. Marie Wilmoth, Tiffany A. Lemley, Caleb O. Tager, Lisa Vonnahme, Kimberly A. TARGET AUDIENCES: Not relevant to this project. PROJECT MODIFICATIONS: Not relevant to this project.
Impacts
The results of this work has lead to an increase in our understanding of the underlying mechanisms regulating the reporductive processes proposed in our original Hatch project. Increasing our understanding of the mechanistic underpinnings of variation in placental function in the pig is likely to lead to better feeding regimens for pregnant sows to optimize fetal growth, the critical step determining the trajectory of postnatal performance. Further, by working to combine two different, but related, expeirmental paradigms for inquiring about mechanisms responsible for deterimining the quality and quantity of fetal growth in lambs has potential to greatly expand the understanding of basic mechanisms at play during gestation. Finally, by continuing to refine our understanding of the nutritional regulation of progesterone catabolism and the potential processes involved we feel we are increasing the avenues that can be pursued to increase fertility in high-producing dairy cows, without negatively impacting milk production.
Publications
- Wilmoth, T. A., J. M. Koch, D. L. Smith and M. E. Wilson. 2009. Estradiol-17 Alters Trophectoderm Proliferation in Pig Embryos. Journal of Animal and Veterinary Advances (In Press).
- Lemley, C. O., K. A. Vonnahme, L. R. Tager, K. M. Krause and M. E. Wilson. 2009. Insulin secretion, cytochrome P450 activity, liver blood flow and progesterone clearance in dairy cows fed a high cornstarch versus a high fiber diet. American Journal of Physiology (Submitted 11/30/09).
- Lemley, C. O., T. A. Wilmoth, L. R. Tager, K. M. Krause, and M. E. Wilson. 2009. Effect of a high cornstarch diet on hepatic cytochrome P450 2C and 3A activity and progesterone half-life in dairy cows. Journal of Dairy Science (In Press)
- O'Neil, M. R., G. P. Lardy, C. O. Lemley, M. E. Wilson, L. P. Reynolds, J. S. Caton, and K. A. Vonnahme. 2009. Estradiol-17b and linseed meal interact to alter visceral organ mass and hormone levels from ovariectomized ewes. Domestic Animal Endocrinology 37:148-158.
- Koch, J. M., T. A. Wilmoth and M. E. Wilson. 2009. Peri-conceptional growth hormone treatment alters fetal growth and development. Journal of Animal Science (In Press)
- Lemley, C. O., J. M. Koch, K. P. Blemings and M. E. Wilson. 2009. Alterations in progesterone catabolic enzymes, CYP2C and CYP3A, in hepatocytes challenged with insulin and glucagon. Journal of Animal and Veterinary Advances 8:39-46.
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Progress 01/01/08 to 12/31/08
Outputs
OUTPUTS: During the reporting period we have continued to collect samples of placental and endometrial tissue from late pregnant (day 70, 90 and 110 of gestation) gilts. We have begun to assay those samples for the expression of a variety of nutrient transporters and stain samples from each feto-placental unit so that we can estimate the density of blood vessels both in the placenta and adjacent endometrium. We also collected blood samples from ewes treated with growth hormone around the time of breeding, a treatment we have previously demonstrated increases lamb birth weight, weaning weight and reduces lamb response to a growth hormone releasing hormone challenge. These samples were collected every week from treatment (week 0) until week 8 of gestation. We then assayed these samples for IGF-I as a proxy for growth hormone to estimate the duration of the growth hormone treatment effect. We have also continued feeding dairy cows diets designed to elicit divergent insulin secretion. From these cows we have collected liver biopsies to estimate the activity of progesterone catabolic enzymes and used continuous infusions to estimate the metabolic clearance rate of progesterone and liver blood flow in response to our dietary treatments. PARTICIPANTS: Matthew Wilson Project Director K. Marie Krause Co-investigator Kim Vonnahme Co-investigator Jill Koch Graduate student Caleb Lemley Graduate student Tiffany Wilmoth Graduate student TARGET AUDIENCES: Not relevant to this project. PROJECT MODIFICATIONS: Not relevant to this project.
Impacts
The impact of these research projects has been significant. Relative to placental function in the pig, we have detailed the expression of a variety of nutrient transporters in the placenta and adjacent endometrium from each feto-placental unit in litters collected from gilts on days 70, 90 and 110 of gestation. Real-time PCR was used to determine mRNA expression of nutrient transporters, including: rBAT-1, FAT-1, FAT-2, yLAT-1, CAT-1, ASCT-1 and GLUT-3. On day 90, ASCT-1 was found to be more highly expressed in the endometrium associated with feto-placental units having a higher placental efficiency. On day 110, individual feto-placental units having a lower placental efficiency tended to have a higher endometrial mRNA expression rBAT-1 and FAT-1. Relative to the duration of the periconceptional treatment of ewes with growth hormone, we determined IGF-I concentrations (as a proxy for growth hormone). For weeks 1, 2, 3 and 4 of gestation, treated ewes had an increased IGF-1 concentration compared to control ewes (3, 5, 4 and 2.5-fold, respectively). Periconceptional treatment with GH causes IGF-1 to remain elevated for the first half of gestation, potentially altering the uterine environment during a crucial developmental period. Relative to the nutritionally mediated regulation of progesterone catabolism, holstein dairy cows (n = 11) were randomly assigned to a high starch or high fiber diet in a cross-over experimental design consisting of two 14 d periods. Dry matter intake, milk yield and milk lactose yield were not different between the two diets. Insulin concentrations were increased by 22% in cows fed the high starch diet, and both cytochrome P450 2C and cytochrome P450 3A activities were decreased by 50%. Liver blood flow was not different between the two diets. Metabolic clearance rate of progesterone tended to be lower in cows fed the high starch diet versus the high fiber diet. The half-life of progesterone was increased in cows fed the high starch diet versus the high fiber diet. In addition, twenty-two Holstein cows were trained to acquire feed from the Calan gate system and randomly assigned to either a high corn starch or high fiber diet in a cross-over experimental. Dry matter intake was similar for the high starch and high fiber diet, as was milk yield and milk lactose yield. Insulin concentrations at the time of liver biopsy were elevated by 47% in cows consuming the high starch diet versus the high fiber diet. Cytochrome P450 2C activity was decreased by 45%, while cytochrome P450 3A activity tended to be lower by 34% in cows consuming the high starch diet. Insulin was elevated by 19% in cows consuming the high starch diet. The half-life of progesterone tended to be longer in cows fed the high starch diet versus the high fiber diet. In summary, cows with elevated insulin concentrations and lower enzyme activity showed a decrease in progesterone clearance without any changes in liver blood flow, dry matter intake or milk yield. This relationship between insulin and enzyme activity may be a useful approach to decrease high rates of progesterone clearance during pregnancy.
Publications
- Wilson, M. E. And L. L. Anderson. 2008. Mechanistic aspects of foetal development relating to postnatal health and metabolism in pigs. In: Managing Prenatal Development to Enhance Livestock Productivity. pp. . Ed: P. Greenwood and A. Bell. FAO-IAEA
- Lemley, C. O., J. M. Koch, K. P. Blemings and M. E. Wilson. 2008. Alterations in progesterone catabolic enzymes, CYP2C and CYP3A, in hepatocytes challenged with insulin and glucagon. Journal of Animal and Veterinary Advances 8:39-46.
- Galbreath, C. W., E. J. Scholljegerdes, G. P. Lardy, K. G. Odde, M. E. Wilson, J. W. Schroeder and K. A. Vonnahme. 2008. Effect of feeding flax or linseed meal on progesterone clearance rate in ovariectomized ewes. Domestic Animal Endocrinology 35:164-169.
- Lemley, C. O., J. M. Koch, K. P. Blemings, K. M. Krause and M. E. Wilson. 2008. Concomitant changes in progesterone catabolic enzymes, cytochrome P450 2C and 3A, with plasma insulin concentrations in ewes supplemented with sodium acetate or sodium propionate. Animal: An International Journal of Animal Bioscience 2:1223-1229.
- Lemley, C. O., S. T. Butler, W. R. Butler and M. E. Wilson. 2008. Short Communication: Insulin Alters Hepatic Progesterone Catabolic Enzymes, Cytochrome P450 2C and 3A, in Dairy Cows. Journal of Dairy Science 91:641-645.
- Wilson, M. E. and J. M. Koch. 2008. Growth hormone modulation of the early uterine environment and conceptus development in sheep. Havemeyer Monograph Series 21:24-26.
- Lemley, C. O., L. R. Tager, K. M. Krause and M. E. Wilson. 2008. Peripheral concentrations of insulin are negatively correlated with cytochrome P450 3A activity and mRNA expression in dairy cows. Journal of Animal Science 86 (Supplement 2):387.
- Wilson, M. E. and J. M. Koch. 2008. The early gestational uterine environment programs placental nutrient transport. Journal of Animal Science 86 (Supplement 1):In Press.
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Progress 01/01/07 to 12/31/07
Outputs
OUTPUTS: Objective IA. Over 80 million Americans are affected by a cardiovascular disease, which is expected to cost $431 billion in 2007. Cardiovascular disease is the number one killer of both men and women in America. Perturbations of fetal growth can lead to a programmed dysfunction of the cardiovascular system resulting in hypertension, heart disease, atherosclerosis, thickening of the aorta, premature stiffening of carotid arteries, etc. Perturbations of fetal growth result from intrauterine growth restriction. Experimentally induced intrauterine growth restriction involves limiting the delivery of nutrients to the fetus. Uteroplacental function can be dramatically altered by altering the growth of the early embryo. We have taken the novel approach of trying to understand how to stimulate fetal growth by modifying the early uterine environment. By modifying the early uterine environment we increased the growth of the embryo, which directly altered the growth and development of
the fetus, leading to an increase in birth weight. Most importantly, there was a marked reduction of the thickness of the left ventricular wall of the heart, leading us to suggest a dramatic alteration in cardiovascular physiology. Objective II. Dairy cow pregnancy rates have declined 30% in the last 50 years, and approximately 80% of the total loss occurs during the embryonic stage (before day 42). Throughout the United States, which has approximately 9 million dairy cows, the industry is losing almost a billion dollars each year due to high embryonic loss. Authors have suggested that lower pregnancy rates result from low progesterone concentrations. Our laboratory has shown that hepatocytes cultured in the presence of insulin exhibit reduced progesterone catabolism, which is a result of lowered hepatic cytochrome P450 activity. Currently, there is a paucity of information on nutritional regulation of cytochrome P450 activity, especially on mechanisms to reduce these catabolic
enzymes. Therefore, a study designed to measure differences in the activity of cytochrome P450s and progesterone clearance, following alterations in diet would contribute substantially to understanding the influence of nutrition on peripheral progesterone concentrations.
PARTICIPANTS: Jill M. Koch, WVU, Graduate Student Caleb O. Lemley, WVU, Graduate Student K. Marie Krause, WVU, Coinvestigator Kenneth P. Blemings, WVU, Coinvestigator Kimberly A. Vonnahme, NDSU, Coinvestigator W. Ronald Butler, Cornell, Coinvestigator Ronald R. Magness, Coinvestigator
TARGET AUDIENCES: Scientists working in the fields of Reproductive Physiology, Dairy Nutrition, Developmental Biology, Developmental Programming, Steroid Biochemistry and Animal Management.
Impacts
Objective IA. Increasing our understanding of how the environment and nutrition of the developing conceptus during pregnancy influences the growth and health of the offspring throughout their lives should lead to modifications in the manner in which we manage food animals and has the potential to improve human health in the future. Objective II. The long term goal of our research is to enhance reproductive efficiency in dairy cows. The objective of the current research is to establish dietary modifications that result in a decrease in progesterone catabolism. Our central hypothesis is that elevated insulin concentrations will decrease expression and activity of cytochrome P450s leading to a reduced clearance rate of progesterone. Therefore our rationale is that a better understanding of nutritional manipulation of cytochrome P450 activity will allow us to elevate progesterone concentrations during vital periods of gestation and in turn decrease embryonic loss.
Publications
- Costine, B. A., E. K. Inskeep, K. P. Blemings, J. A. Flores and M. E. Wilson. 2007. Mechanisms of luteal resistance to prostaglandin F2α during maternal recognition of pregnancy in ewes. Domestic Animal Endocrinology 32:106-121.
- Wilson, M. E. 2007. The role of the conceptus in determining placental efficiency and the capacity of the uterus. In: Manipulating Pig Production XI. pp. 128-135 Ed: J. E. Paterson and J. A. Barker. Australasian Pig Science Association Inc., Werribee, Victoria, Australia.
- Koch, J. M., J. S. Moritz, D. C. Lay and M. E. Wilson. 2007. Effect of melengestrol acetate as an alternative to induce molting in hens on the expression of yolk proteins and turnover of oviductal epithelium. Animal Reproduction Science 102:14-23.
- Koch, J. M., D. C. Lay Jr., K. A. McMunn and M. E. Wilson. 2007. Alternative method to induce molt that addresses hen well-being. Poultry Science 86:614-620.
- Dixon, A. B., M. Knights, J. L. Winkler, D. J. Marsh, J. L. Pate, M. E. Wilson, R. A. Dailey, G. Seidel and E. K. Inskeep. 2007. Patterns of and factors affecting late embryonic and fetal mortality in sheep. Journal of Animal Science 85:1274-1284.
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Progress 01/01/06 to 12/31/06
Outputs
We have made significant progress on two of our three aims (Objectives IA and II). We will begin to collect data on Objective IB this fall. Objective IA: In addition to the difference in birth weight that we had previously reported, we have found that lambs born to ewes treated with growth hormone at breeding, to alter the environment to which the embryo is exposed, have an improved development of the cardiovascular system and an altered response from their brain-liver endocrine axis. These data are quite exciting as they indicate that the environment an early embryo is exposed can have long term consequences for the growth and health of the individual. We have also recently initiated studies to investigate what might be different about the development of the very early embryo in response to the treatment with growth hormone. In a limited number of animals, we find that embryos from ewes treated with growth hormone have a 33 percent increase in the ratio of
trophectoderm to inner cell mass. We think that these data will ultimately lead to alterations in the way we feed and manage animals around the time of breeding to improve growth and health in the offspring. Furthermore, these data may translate to interventions applicable to pregnant humans to improve the health and well-being of generations to come. Objective II: We have recently completed two replicates of an experiment in which we supplement ewes' rations with two different fatty acids in an isocaloric manner. This supplementation results in very different patterns of insulin secretion following feeding. Specifically, when ewes are supplemented with propionate (a gluconeogenic substrate) there is a dramatic rise in insulin following feeding. Coincident with the increase in insulin following propionate supplementation, there is a dramatic reduction (by one-half) in the activity of the two enzymes that catabolize progesterone and not surprisingly a marked reduction in the metabolic
clearance rate of progesterone. This sort of nutritional approach to decreasing progesterone catabolism has the potential to have a tremendous impact on the dairy industry where pregnancy wastage is an enormous economic burden.
Impacts
The impact of understanding how improving the environment the early embryo is exposed to improves their postnatal growth and health would be a tremendous boon to both livestock and humans. Specifically, our approach appears to improve the development of the cardiovascular system and alter the responsiveness of the growth hormone axis. Additionally, if we can continue to gain insight into the metabolic regulation of progesterone catabolism then we ought to be able to dramatically improve the fertility of livestock species, particularly the lactating dairy cow.
Publications
- Peer-reviewed papers Wilson, M. E. And L. L. Anderson. 2006. Mechanistic aspects of foetal development relating to postnatal health and metabolism in pigs. In: Managing Prenatal Development to Enhance Livestock Productivity. pp. . Ed: P. Greenwood and A. Bell. FAO-IAEA
- Wilson, M. E. and J. M. Koch. 2006. Growth hormone modulation of the early uterine environment and conceptus development in sheep. Havemeyer Foundation Workshop on Embryonic and Fetal Nutrition, May 29th-June 1st, Ravello, Italy.
- Wilson, M. E. and J. M. Koch. 2007. Growth hormone modulation of the early uterine environment and conceptus development in sheep. Havemeyer Monograph Series (In Press).
- Smith, D. L., B. M. Stinefelt, K. P. Blemings and M. E. Wilson. 2006. Diet-induced alterations in progesterone clearance appear to be mediated by insulin signaling in hepatocytes. Journal of Animal Science 84:1102-1109.
- Abstracts: Koch, J. M. and M. E. Wilson. 2007. Effects of sustained release growth hormone on lambs at birth. Proceedings of the Society for Gynecologic Investigation In Press.
- Wilmoth, T. A., C. O. Lemley, J. M. Koch, K. P. Blemings and M. E. Wilson. 2007. Validation of a ruminant hepatocyte culture for assaying the activity of enzymes responsible for progesterone catabolism, cytochrome P450 (CYP) 2C and 3A Journal of Animal Science 85 (Supplement 2):In Press.
- Koch, J.M., C.O. Lemley, M.E. Wilson. 2007. Effects of Growth Hormone at Breeding on the offspring's insulin-like growth factor-I (IGF-I) response to growth hormone releasing hormone challenge. Journal of Animal Science 85 (Supplement 2):In Press.
- Koch, J. M., B. A. Costine and M. E. Wilson. 2006. Effect of growth hormone given at breeding on amino acid and glucose transporters in the sheep placenta. Biology of Reproduction 75 (Supplement 1):108.
- Lemley, C. O., J. M. Koch, K. P. Blemings and M. E. Wilson. 2006. Insulin induced alteration in progesterone catabolism in murine hepatocytes appears to be mediated by CYP3A13. Biology of Reproduction 75 (Supplement 1):159.
- Lemley, C. O., J. M. Koch, K. P. Blemings and M. E. Wilson. 2007. Determination of insulin concentrations in sheep isocalorically supplemented with two different volatile fatty acids. Journal of Animal Science 85 (Supplement 2):In Press.
- Wilson, M. E. and J. M. Koch. 2006. Growth hormone modulation of the early uterine environment and conceptus development in sheep. Handbook of Abstracts for the Havemeyer Foundation Workshop on Embryonic and Fetal Nutrition II:11.
- Wilmoth,T. A., D. L. Smith, J. M. Koch and M. E. Wilson. 2006. Effects of estradiol on the uterine environment and trophectoderm in the gilt. Journal of Animal Science 84 (Supplement 2):53.
- Koch, J. M. and M. E. Wilson. 2006. Effects of sustained release growth hormone on lambs at birth. Journal of Animal Science 84 (Supplement 2):128.
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