Endometritis in dairy cows is a major global issue and has been associated with a decrease in reproductive performance. The aim of this study was to quantify the effect of endometritis (as defined by the presence of any abnormal vaginal discharge after 21 days post-partum) on post-partum ovarian cyclicity in dairy cows. Milk progesterone analysis was used to monitor reproductive cyclicity in 170 dairy cows across three different commercial herds. Associations between the occurrence of endometritis and the incidence risk of a variety of atypical cycle profiles during the calving to conception period were investigated to establish the importance of endometritis on post-partum ovarian activity. Endometritis increased the incidence odds of atypical ovarian profiles (P < 0.05) with prolonged luteal activity being the most affected (P < 0.05), but also showed prolonged time (3 days) to onset of luteal activity after parturition (P < 0.05). Using milk progesterone analysis, we found a relatively low incidence odds for reproductive cycle problems in healthy cows during the calving to conception period. However, the incidence odds of cycle problems, in particular prolonged luteal activity, were high in cows that had experienced endometritis, which would have significantly impaired reproductive function.

Despite the importance of progesterone on the fertility of lactating dairy cows, the factors that affect post ovulatory progesterone concentration are still unclear. Thus, the aim of the present study was to identify factors associated with the post ovulatory progesterone rise following 1st insemination in lactating dairy cows. Data collected across a number of complimentary studies were compiled to produce a single database of 168 lactating Holstein Friesian dairy cows maintained under commercial conditions. In all animals a number of variables were measured during the insemination period and related to milk progesterone measured on day 5 following 1st artificial insemination (AI). Overall, 44% of cows conceived to 1st AI and while mean day 5 progesterone was not significantly higher in these cows, there was a significant quadratic relationship between milk progesterone concentration and conception rate. While a number of variables showed some association with progesterone concentration, the only variable showing a strong and repeatable relationship was plasma leptin concentration. We conclude that adequate but not excessive progesterone levels on day 5 bring about a better fertility, and plasma leptin concentration may be a much better indicator of metabolic status in lactating dairy cows.

Reproductive management in cattle requires the synchrony of follicle development and oestrus before insemination. However, ovulation of follicles that have not undergone normal physiological maturation can lead to suboptimal luteal function. Here, we investigated the expression of a targeted set of 47 genes in (a) a first-wave vs final-wave dominant follicle (DF; the latter destined to ovulate spontaneously) and (b) 6-day-old corpora lutea (CLs) following either spontaneous ovulation or induced ovulation of a first-wave DF to ascertain their functional significance for competent CL development. Both the mass and progesterone-synthesising capacity of a CL formed following induced ovulation of a first-wave DF were impaired. These impaired CLs had reduced expression of steroidogenic enzymes (e.g. STAR and HSD3B1), luteotrophic receptors (LHCGR) and angiogenic regulators (e.g. VEGFA) and increased expression of BMP2 (linked to luteolysis). Relative to final-wave DFs, characteristic features of first-wave DFs included reduced oestradiol concentrations and a reduced oestradiol:progesterone ratio in the face of increased expression of key steroidogenic enzymes (i.e. CYP11A1, HSD3B1 and CYP19A1) in granulosa cells and reduced expression of the HDL receptor SCARB1 in thecal cells. Transcripts for further components of the TGF and IGF systems (e.g. INHA, INHBA, IGF2R and IGFBP2) varied between the first- and final-wave DFs. These results highlight the importance of hormones such as progesterone interacting with local components of both the TGF and IGF systems to affect the maturation of the ovulatory follicle and functional competency of the subsequent CL.

In light of increasing global protein prices and with the need to reduce environmental impact of contemporary systems of milk production, the current review seeks to assess the feasibility of reducing levels of dietary CP in dairy cow diets. At CP levels between 140 and 220 g/kg DM there is a strong positive relationship between CP concentration and dry matter intake (DMI). However, such effects are modest and reductions in DMI when dietary CP is below 180 g/kg DM can be at least partially offset by improving the digestibility and amino acid profile of the undegradable protein (UDP) component of the diet or by increasing rumen fermentable energy. Level and balance of intestinally absorbable amino acids, in particular methionine and lysine, may become limiting at lower CP concentrations. In general the amino acid composition of microbial protein is superior to that of UDP, so that dietary strategies that aim to promote microbial protein synthesis in the rumen may go some way to correcting for amino acid imbalances in low CP diets. For example, reducing the level of NDF, while increasing the proportion of starch, can lead to improvements in nitrogen (N) utilisation as great as that achieved by reducing dietary CP to below 150 g/kg. A systematic review and meta-analysis of responses to rumen protected forms of methionine and lysine was conducted for early/mid lactation cows fed diets containing ⩽150 g CP/kg DM. This analysis revealed a small but significant (P=0.002) increase in milk protein yield when cows were supplemented with these rumen protected amino acids. Variation in milk and milk protein yield responses between studies was not random but due to differences in diet composition between studies. Cows fed low CP diets can respond to supplemental methionine and lysine so long as DMI is not limiting, metabolisable protein (MP) is not grossly deficient and other amino acids such as histidine and leucine do not become rate limiting. Whereas excess dietary protein can impair reproduction and can contribute to lameness, there is no evidence to indicate that reducing dietary CP levels to around 140 to 150 g CP/kg DM will have any detrimental effect on either cow fertility or health. Contemporary models that estimate MP requirements of dairy cows may require refinement and further validation in order to predict responses with low CP diets.

Cyclic ewes were treated with control vehicle or progesterone receptor antagonist (onapristone; 100mg i.m. twice daily) during either early (day 3-5) or late (day 12-14) luteal phase and plasma samples collected for hormone analysis and to determine endogenous and oxytocin induced PGF2α release. On day 14 and 17, ewes were euthanised and reproductive tracts collected for ovarian morphology and endometrium for oxytoxin and steroid hormone receptor analysis. Early treatment increased LH, but not progesterone or oestradiol, while late treatment elevated all three hormones. Early treatment delayed the up-regulation of endometrial oxytocin receptors and responsiveness to oxytocin challenge, delaying luteolysis. Late treatment advanced development of oxytocin receptors and responsiveness to oxytocin though not timing of luteolysis. Patterns of hormone receptor mRNA were differentially disrupted by treatments. Results provide mechanistic insight into hormonal control of the oestrous cycle and identify the ability of the luteolytic mechanism to dissociate from functional luteolysis.

Plasma progesterone concentrations were estimated for 381 nulliparous dairy heifers from eight farms on days 4–6 after insemination. Heifers were synchronised using two treatments of prostaglandin F2α (PGF2α) 11 days apart. Alternate heifers were treated with 1500 IU human chorionic gonadotrophin (hCG) at the time of sampling, and the pregnancy status assessed ∼35 days after insemination. Heifers with no corpus luteum (CL; n‐30) at the second PGF2α had lower progesterone concentrations (P<0.001) and pregnancy rates (PR; P=0.001) compared with heifers with CL (n=351, defined as cyclic). The risk of no CL increased in heifers <15 months of age (OR=7.5, P<0.001) and with body condition score <2.5 (OR=4.5, P=0.001). Those with no CL were removed from further analysis. The 20 per cent cyclic control heifers with progesterone concentrations ≤2 ng/ml on days 4–6 had lower PRs compared with those with higher concentrations (42 per cent vs 64 per cent, OR=0.3, P=0.01). There was no overall effect of hCG on PR compared with controls (51 per cent vs 60 per cent, P=0.10). However, heifers with high progesterone on days 4–6 (>2 ng/ml) had a lower PR after treatment (51 per cent vs 64 per cent, OR=0.6, P=0.02). Heifers inseminated with sexed (n=18) compared with conventional (n=333) semen had a decreased PR (33 per cent vs 57 per cent, OR=0.2, P=0.01). This study demonstrates that higher plasma progesterone concentrations as early as days 4–6 after insemination have a positive effect on first insemination PR, but there was no beneficial effect of hCG treatment on these days in dairy heifers.

Inadequate luteal support of early embryo development remains an important cause of embryo mortality. However, many of the postulated causes of this luteal inadequacy such as high milk yield are not supported by current scientific evidence and the true causal factors remain elusive. Both direct progesterone supplementation and the enhancement of endogenous progesterone production through gonadotrophic stimulation can improve pregnancy rates. However, the results of such approached have proved inconsistent and careful attention must be paid to the timing of application of these therapies.

One of the many important factors associated with the poor pregnancy rates seen in modern dairy cows is the failure of the embryo to produce sufficient interferon tau (IFN-tau), the protein needed to prevent luteolysis and thus establish the pregnancy. The key factor responsible for this is the failure of the embryo to develop sufficiently well due to inadequate production of progesterone by the corpus luteum. Attempts to establish the mechanisms through which progesterone production is impaired have proved challenging but a current solutiuon is available in the form of progesterone supplementation. In order to maximise benefits it is important that this treatment is given to the correct cows at the correct time. Detailed analysis of a number of supplementation trials, supported by a series of studies on the hormonal control of embryo development, have revealed that a 10% improvement in pregnancy rate can be achieved by administering progesterone during the period of the post ovulatory rise, around day 5 following insemination. Treatment at other times yielded little if any real benefit.

在过去的20年内,英国乳品加工业中奶牛的繁殖力在下降,初配牛的产犊率由60%降至40%,平均每年下降1%.同时繁殖障碍的发生率却大大增加.奶牛成功受孕需要胚胎的充分发育,从而产生足量的抗黄体溶解干扰素(IFN)-π.

We have investigated the effects administering exogenous progesterone, via insertion of a controlled internal drug release (CIDR) for 4 days, from either day 5 or day 12 of the oestrous cycle on plasma oestradiol concentrations. In study 1, in which progesterone was administered from day 5, measurement of plasma oestradiol in daily samples revealed a significant (p < 0.001) decrease in peripheral oestradiol concentration. In contrast, in study 2, similar administration of progesterone from day 12 had no effect on plasma oestradiol concentration. In study 3, collection of hourly samples following progesterone treatment on day 5 revealed peak progesterone concentrations within 1 h of CIDR insertion and nadir oestradiol concentrations within 4 h. The results demonstrate that treatment with progesterone early in the luteal phase causes a rapid inhibition of oestradiol secretion, while later treatment does not. While improvements in pregnancy rate following progesterone treatment at this time have traditionally been attributed to increases in progesterone, the potential involvement of decreased oestradiol secretion has often been overlooked.

The feeding of conjugated linoleic acid (CLA) supplements to early-lactation dairy cows has been shown to decrease milk fat synthesis and possibly improve reproductive performance. However, previously reported studies used too few animals to clearly establish the effect of CLA on reproduction. Our objective was to combine data from these studies to evaluate the association of CLA with time to first ovulation and time to conception using methods of survival analysis and overall success of pregnancy by logistic regression. A database was compiled of individual animal data (n = 212) from 5 controlled studies in which CLA had been supplemented to early-lactation dairy cows. Survival analysis incorporated both semi-parametric models (Cox proportional hazards) and parametric models (log-normal). The probability of cows becoming pregnant increased in a nonlinear manner as trans-10, cis-12 CLA dose increased, with the optimal dose predicted to be 10.1 g/d. At the optimal dose, the probability of pregnancy was increased by 26% compared with those animals receiving no CLA (probability = 91% and 72%, respectively). Similarly, the log-normal model predicted that time to conception was decreased in a nonlinear manner with increasing trans-10, cis-12 CLA dose. The predicted optimal dose was 10.5 g of trans-10, cis-12 CLA/d and at this dose the median time to conception was decreased by 34 d when compared with those cows not receiving CLA (117 vs. 151 d in milk, respectively). The log-normal model was also the best-fit model for time to first ovulation. Overall, this multi-study analysis demonstrated a strong concordance between the nature of the dose response and the predicted optimal dose of trans-10, cis-12 CLA across the 3 reproductive variables evaluated. These results indicate that reproductive performance of dairy cows may be improved by feeding of CLA supplements during early lactation.

The incidence of low day 5 milk progesterone in dairy cows has been investigated and the efficacy of treating the problem assessed. The incidence of inadequate milk progesterone (empirically defined as <3 ng/mL) in repeat breeder cows was 34% compared with 11.4% in first insemination cows. Treatment with an intravaginal progesterone device for 7 days starting from day 5 or 6 did not improve pregnancy rate. Treatment with 1500 iu human chorionic gonadotrophin (hCG) on day 5 gave an increase in pregnancy rate that was dependent on initial progesterone concentration and significant (P < 0.05) in multiparous but not primiparous cows. While the incidence of inadequate day 5 progesterone was high in repeat breeder cows, it was responsive to hCG treatment, although only in multiparous and not primiparous animals.

Delayed resumption of postpartum oestrous cycles in dairy cows is linked to low plasma insulin status and associated changes in ovarian function. Previous studies showed that plasma insulin can be raised by increasing dietary starch concentration, but it is not known if rumen digestible starch and bypass starch would be equally effective. The objective of the two experiments reported here was to investigate whether site of dietary starch digestion influences metabolic hormones and ovarian function. Cows were fed on a standard diet from calving until 40 days postpartum and then fed on treatment diets until 70 days postpartum. In Experiment 1, six cows were transferred to each of five diets with a starch content of 190 g/kg DM, of which 0.40, 0.45, 0.50,0.55 or 0.60 was bypass starch. In Experiment 2, eight cows were transferred to each of four diets containing concentrates with low (L) or high (H) starch content and predominantly grass (G) or maize (M) silages in a factorial design. Between 60 and 70 days postpartum, plasma insulin was not influenced by site of starch digestion in Experiment 1, but insulin was increased by H (P = 0.020) and M (P = 0.048) in Experiment 2. Dietary bypass starch content negatively influenced plasma glucagon concentration (P = 0.046) and the insulin to glucagon ratio (P = 0.005) in Experiment 1. Growth hormone, insulin-like growth factor-I and leptin did not vary among diets in either experiment. The number of small (<5 mm) ovarian follicles before ovulation tended (P = 0.056) to decrease with increasing dietary bypass starch content and was negatively related (P = 0.046) to plasma glucagon in Experiment 1, but there was no other dietary effect on ovarian function in either experiment. The main conclusion from these experiments is that rumen digestible starch and rumen bypass starch can be equally effective for maintaining plasma insulin and ovarian function of high-yielding dairy cows in early lactation. (C) 2009 Elsevier B.V. All rights reserved.

Dietary stimulation of insulin in post partum dairy cows has been found to enhance ovarian follicle development but to impair oocyte developmental competence. The objective of this study was to test the hypothesis that pregnancy rate would be improved by feeding a diet to stimulate higher insulin (H) until cows resumed ovarian cyclic activity after parturition, and then feeding a diet to lower insulin (L) during the mating period. Each diet was fed to 30 post partum dairy cows until their first rise in milk progesterone, when 15 cows in each group were transferred to the other diet (treatments HL and LH) and 15 cows in each group remained on their original diet (treatments HH and LL) until 120 days post partum. Treatments did not affect dry matter intake, milk yield and metabolisable energy balance. Plasma insulin concentration was elevated in cows fed on H compared with cows fed on L. Treatment did not affect days to first progesterone rise, first oestrus or first insemination. At 120 days post partum, 27% of cows on each of treatments HH, LL and LH were pregnant, but 60% of cows on treatment HL were pregnant (P=0.021). These findings support the concept that physiological relationships between insulin and the reproductive system vary according to stage of the reproductive cycle, and suggest that pregnancy rate can be enhanced by a two-diet strategy tailored to optimise responses before and after the first post partum ovulation.

Ovarian function is dependent on the establishment and continual remodelling of a complex vascular system. This enables the follicle and/or corpus luteum (CL) to receive the required supply of nutrients, oxygen and hormonal support as well as facilitating the release of steroids. Moreover, the inhibition of angiogenesis results in the attenuation of follicular growth, disruption of ovulation and drastic effects on the development and function of the CL. It appears that the production and action of vascular endothelial growth factor A (VEGFA) is necessary at all these stages of development. However, the expression of fibroblast growth factor 2 (FGF2) in the cow is more dynamic than that of VEGFA with a dramatic upregulation during the follicular-luteal transition. This upregulation is then likely to initiate intense angiogenesis in the presence of high VEGFA levels. Recently, we have developed a novel ovarian physiological angiogenesis culture system in which highly organised and intricate endothelial cell networks are formed. This system will enable us to elucidate the complex inter-play between FGF2 and VEGFA as well as other angiogenic factors in the regulation of luteal angiogenesis. Furthermore, recent evidence indicates that pericytes might play an active role in driving angiogenesis and highlights the importance of pericyte-endothelial interactions in this process. Finally, the targeted promotion of angiogenesis may lead to the development of novel strategies to alleviate luteal inadequacy and infertility.

Ovarian function is dependent on the establishment and continual remodelling of a complex vascular system. This enables the follicle and/or corpus luteum (CL) to receive the required supply of nutrients, oxygen and hormonal support as well as facilitating the release ofsteroids. Moreover, the inhibition ofangiogenesis resultsin the attenuationoffolliculargrowth, disruption ofovulation and drastic effects on the development and function of the CL. It appears that the production and action of vascular endothelial growth factor A (VEGFA) is necessary at all these stages of development. However, the expression offibroblast growth factor 2 (FGF2) in the cow is more dynamic than that of VEGFA with a dramatic upregulation during the follicular-luteal transition. This upregulation is then likely to initiate intense angiogenesis in the presence of high VEGFA levels. Recently, we have developed a novel ovarian physiological angiogenesis culture system in which highly organised and intricate endothelial cell networks are formed. This system will enable us to elucidate the complex inter-play between FGF2 and VEGFA as well as other angiogenic factors in the regulation of luteal angiogenesis. Furthermore, recent evidence indicates that pericytes might play an active role in driving angiogenesis and highlights the importance of pericyte-endothelial interactions in this process. Finally, the targeted promotion of angiogenesis may lead to the development of novel strategies to alleviate luteal inadequacy and infertility. Reproduction (2009) 138 869-881