Studies relating to Polyunsaturated Fatty Acid (PUFA) Supplementation and Fertility in Cattle

End of project reportReproductive inefficiency has a significant impact on the economic performance of both dairy and beef herds, particularly in seasonal calving systems. Nutrition plays a fundamental role in reproduction. Furthermore, there is emerging evidence that supplemental dietary polyunsaturated fatty acids (PUFA) may increase cow fertility independent of their role as energy substrates. A number of studies have reported enhanced reproductive performance in dairy cows following dietary supplementation with sources of n-3 PUFA. However the possible mechanisms involved have not been identified and there is some inconsistency in the published literature on this topic. The objective of the research reported was to conduct a holistic examination of the effects of dietary long-chain n-3 PUFA supplementation on metabolic and reproductive responses in cattle. Such information is essential for the appropriate formulation of diets to enhance cow reproductive performance and in particular embryo survival

Studies relating to protein expression in the uterus of the cow

End of project reportEmbryo loss is a major cause of reproductive wastage in the cow. The majority of embryo loss occurs in the first 16 days after fertilisation when the embryo is critically dependent on the maternal uterine environment for survival. Despite the central role of uterine fluid in the normal growth and development of the embryo, there is limited information on the protein composition of these fluids. The main objectives of the studies in this thesis were to examine the protein composition of the bovine uterus during the oestrous cycle and to examine the relationship between the concentration of systemic progesterone and uterine protein expression in the cow. In the first study, the concentration of retinol-binding protein (RBP) in the bovine uterus was found to vary across the cycle and was 5-15-fold higher (P0.05) of the cycle, which was surprising given that previous studies have indicated that uterine RBP gene expression was positively associated with the concentration of systemic progesterone. In the second study, IGF binding protein 2 (IGFBP-2), IGFBP-3, IGFBP-4 and IGFBP-5 were identified in uterine fluid on days 3, 7, 11 and 15 of the oestrous cycle. There was a local effect on the concentration of IGFBPs where the concentration was greater on the ipsilateral side than that on the contralateral side for IGFBP-2 (P<0.05), 3 (P<0.01) and 5 (P<0.01) on day 15. This difference is a further indication of a local controlling mechanism regulating proteins between the uterine horns. Similar to RBP expression this study could find no significant relationship between the concentration of systemic progesterone and IGFBP concentrations on Day 7 of the oestrous cycle. In the third study, changes in the global pattern of uterine proteins between Days 3 and Day 15 of the oestrous cycle were examined using two-dimensional electrophoresis (2-DE). Six proteins were found to be upregulated on Day 15 compared to Day 3. Three proteins of these were identified as aldose reductase, plakoglobin and heat shock protein 27 while the other three proteins were identified as bovine serum albumin. Aldose reductase, an enzyme directly involved in the production of sorbitol and indirectly of fructose, was 10-fold higher (P<0.0001) on Day 15 compared to Day 3. Plakoglobin (Pg) was upregulated 2.3-fold (P<0.0001) on Day 15 compared to Day 3. Pg is a component of cellular junctions and its up-regulation may have a role in the uterine glandular epithelium. Heat shock protein 27 (Hsp27) was higher on Day 15 than on Day 3 (P<0.01) and Hsp27 was 1.4-fold higher in the ipsilateral compared to the contralateral uterine horn (P<0.01). Hsp27 may be secreted in response to potential stresses in the uterus or act as a molecular chaperone. On Day 7 there was no difference (P<0.05) in the pattern of proteins secreted between cows with low (2.7±0.10ng/ml) and high (4.8±0.13 ng/ml) concentrations of systemic progesterone on Day 7. The results of these studies have shown that dramatic changes occur in protein expression across the bovine oestrous cycle. Additionally, it emphasises the need for gene studies to be followed with protein studies as an adjunct or complementary tool. Proteins have a wide range of essential roles in the uterus and together these studies provide novel information on protein expression in the uterus of the cow

Biotechnology in Cattle Reproduction.

End of Project ReportTeagasc acknowledges support from the European Union 3rd and 4th Framework Programmes (CT-92-0163 and CT-95-0190)Over the next decade the Irish agri-food industry will have to compete in a rapidly changing world environment arising from increased competitiveness, decreased world market prices and increased consumer demands for higher quality, healthier and safer food. To become competitive in this environment the scale and efficiency of production at both farm and factory level will have to increase significantly and this must be achieved with due regard for the protection of the environment and the welfare of animals. New technologies will be needed to achieve this. Biotechnology will be central to the development of these new technologies. This project has been concerned with the identification and evaluation of biotechnology developments that have the potential to increase reproductive efficiency in cattle. This includes a range of technologies relating to the in vitro production, manipulation, cryopreservation and transfer of cattle embryos. The potential of other emerging technologies such as embryo and sperm sexing, cloning and biopharming or the production of commercially desirable proteins in cows milk are also addressed in this report.European Unoi

Protein Nutrition and Fertility in Cattle.

End of Project ReportThe objectives of the project were to determine the effects of dietary crude protein intake, on blood concentrations of ammonia and urea and on fertility, and on the possible biological mechanisms involved. Heifers were used in all experiments in order to avoid any confounding effects of lactation and, or, negative energy balance. The results are summarised as follows. • • • • 5 Blood urea and ammonia increased linearly with increases in dietary urea. Fermentable carbohydrate, in the form of molassed sugar beet pulp (MSBP), was more effective than barley in reducing blood urea concentration. Plasma ammonia concentrations were more variable but MSBP was again more effective than barley in reducing these. Embryo survival rate was not affected by either dietary crude protein (CP) or fermentable energy intake irrespective of whether animals were fed a silage diet supplemented with feed grade urea or were grazing pasture with a high crude protein content. Despite generating systemic urea concentrations of up to 25 mmol/L and systemic ammonia concentrations of up to 242 μmol/L no significant relationship between these parameters and embryo survival rate was established. Consistent with these results on embryo survival, elevated blood urea or ammonia had little effect on the biochemical composition of oviduct fluid, the environment of the early developing embryo. It can be concluded that elevations in systemic concentrations of ammonia or urea per se, particularly of the magnitude observed under normal feeding conditions, are unlikely to impair embryo survival in cattle as a consequence of disruptions to the oviductal environment

Viability of in vitro produced cattle embryos.

End of Project ReportEmbryo transfer is being increasingly used in the cattle industry. As well as direct embryo transfers, many embryo-based biotechnologies have the potential to improve cattle production efficiency through enhanced breeding strategies, by facilitating the introduction of desirable traits such as disease resistance and through the production of desirable medical or pharmaceutical products in the milk. These biotechnologies are, however, dependent on a supply of viable in vitro produced (IVP) embryos. While the in vitro fertilization rate is high (80%) in cattle, only about 30 transferable embryos, or blastocysts, are produced from every 100 fertilized oocytes. A major factor affecting the viability of IVP embryos is their failure, in a high proportion of cases, to undergo normal development to the blastocyst stage in the manner of in vivo embryos. The major problem relates to a failure of the cells of IVP embryos to form a compact cell mass when they are 5 - 6 days old. This ultimately leads to developmental problems and compromised viability. Cell compaction is recognized as a critical event in early embryo development and has been associated with marked changes in protein synthesis and phosphorylation in the embryos of some species. This report is the first, to our knowledge, to describe the rate and pattern of protein synthesis and phosphorylation before, during and after compaction in both in vivo and in IVP cattle embryos. The main results are summarised below.European Union 4th Framework Programmes (Contract CT-95-0032

Fluctuations in Energy Intake and Fertility in Cattle.

End of Project ReportsReproductive failure in dairy cows results in fewer calves born, lower milk sales, slower genetic progress and consequently, significant financial loss to the industry. Dairy cattle breed improvement programmes have, at least until very recently, focused primarily on increasing the yields of milk or milk solids. The resulting genetic improvement has led to significant increases in milk yield per cow but this increase is now associated with a significant decline in cow reproductive wastage. An important part of the Teagasc research programme in this area is to determine the time at which embryo loss occurs and also to determine whether the extent of the embryo loss is affected by the energy nutrition of the cow and to devise strategies to reduce its extent. This project has focused on the relationship between changes in dietary energy intake near the time of insemination and the extent and pattern of embryo survival. The main results are summarised in this report and detailed results of the several experiments involved have been published in the papers listed at the end of this report. • • 4 The objectives of this project were to determine the effect of changes in energy intake near the time of insemination on embryo loss rate, on the timing of embryo loss and on the possible biological mechanisms involved. Cross bred heifers were provided with either high or low energy intakes that were based on pasture allowances calculated to provide either 0.8 or 2.0 times their maintenance requirements. These energy intakes were allocated for two weeks before and about five weeks after insemination. The effect of the changes in energy intake on embryo loss and on the time at which embryo loss occurred, relative to the time of insemination, was established. Possible associations between embryo loss and blood concentrations of progesterone, NEFAs, insulin and glucose were examined. A sudden reduction from a high to a low energy intake imposed for two weeks from the day of insemination reduced the subsequent embryo survival rate by 30 percentage points to a survival rate of 38%. When energy intake over this same period was either maintained or increased, embryo survival rate remained high (overall mean, 69%), within a range of 65-71%. The time at which embryo loss occurred was established. Embryo survival or pregnancy rates measured on days 14 and 30 after insemination and at full term were 68%, 76% and 72%, respectively. These results provide new information indicating that most embryo loss, at least in heifers, had occurred on or before day 14 after insemination. There was no evidence of any association between the shortterm changes in energy intake either before or after AI and blood progesterone concentration. Neither was there any evidence that the detrimental effect of the sudden reduction in energy intake on embryo survival was mediated through changes in the systemic concentrations of non-esterified fatty acids (NEFAs) or insulin. There was a suggestion, however, that the detrimental effect of the reduced energy intake may operate through a reduction in systemic glucose concentrations.Dairy Levy Farmer Fun

Increasing the use of AI in suckler herds.

End of Project ReportReproductive efficiency is a major factor affecting production and economic efficiency of beef herds. For herds using artificial insemination (AI) heat detection rate and calving rate are the two major determinants of compactness of calving and ultimately the calving-to-calving interval. Heat detection is a time consuming repetitive chore that must be carried out up to 5-times each day for as long as AI is used. Heat detection rate, usually measured as submission rate, is hugely variable from herd-toherd but for most herds only between 40% and 70% of cows that exhibit heat are actually detected by the stockman. Despite an increased understanding of the endocrine control of the oestrous cycle the goal of fixed-time insemination is not yet consistently achievable in either cows or heifers treated at different stages of the oestrous cycle and in different physiological states. The overall objective of the this project was to develop an improved cost effective hormonal method to control the time of ovulation to allow beef cows be bred by AI without the need for heat detection. A total of 3 studies were carried out and the results are summarised in this report.Department of Agriculture, Food and the Marin

Nutrition and Oestrus and Ovarian Cycles in Cattle

End of Project ReportThe overall objective of this project was to establish the effects of both long- and short-term changes in nutrition on ovarian follicle dynamics and on the systemic concentrations of metabolic and reproductive hormones. In order to avoid the confounding effects of lactation, suckling and maternal–calf bonding, beef heifers were used in a series of three studies.European Union 3rd Framework Programme (Contract AIR3-CT94-1124

Shortening the interval to resumption of ovarian cycles in postpartum beef cows.

End of Project Report• In beef cows the interval from calving to first ovulation, or postpartum interval, is affected by nutrition and by the suckling effect of the calf. • The suckling effect is the biggest determinant of this interval, comprising: (i) physical contact and (ii) maternal bonding between cow and calf. Restricted suckling and calf isolation induce a rapid resumption of oestrous cycles. • Prepartum nutrition is the next most important determinant of the postpartum interval. Cows that calve in poor body condition have a longer interval than cows that calve in good body condition. • Increasing the level of nutrition in the postpartum period has only a limited effect in shortening the postpartum interval. • In most cows the first postpartum ovulation is silent and is succeeded by a short oestrous cycle of approximately 8-10 days. The first observed oestrus occurs prior to the second ovulation. • When used in combination with calf isolation and restricted suckling, progesterone pre-treatment for 6 days induces oestrus in the majority of cows and eliminates the short oestrous cycle. • The prolonged postpartum interval in beef cows is not due to failure of ovarian follicle development but to failure of successive dominant follicles to ovulate due to the inadequate frequency of LH pulses.European Union 3rd Framework Programme (Contract AIR3-CT94-1124

Control of Ovulation Rate in Beef Cattle.

End of Project ReportUnder intensive production systems, the greatest potential for effecting increases in production and economic efficiency in the beef cow herd lies in the possibility of increasing the frequency of twin births. Embryo transfer is technically a successful method of inducing twin births in cattle. While an embryo transfer approach is too costly to allow commercial twinning, it has been used to show that ovulation rate and not uterine capacity is the limiting factor in increasing the reproductive rate of the cow. While ovulation of one or more viable oocytes is central to normal reproduction, knowledge of the control of ovulation and of folliculogenesis on which ovulation depends, is limited. In spite of the fact that many follicles are subjected to the same endogenous hormonal environment and theoretically should all be capable of ovulating, only a tiny proportion do. While gonadotrophic hormones play a central role in ovarian follicle development and ovulation, their action at the ovarian level seems to be controlled by intra-ovarian factors. This intra-ovarian control of ovulation is thought to be exerted partly by the hormone inhibin and partly by other, as yet, unidentified compounds in follicular fluid. This project focused on identification and isolation of ovarian compounds involved in the control of ovulation rate, followed by immunisation against these compounds in order to study the effect on ovulation and the twin calving rates. The main results are summarised here and detailed results have been published in the papers listed at the end of this report