Reliable exclusion of prognostically significant coronary disease in left ventricular dysfunction by cardiac MRI.

AIM: To assess the ability of cardiac magnetic resonance (CMR) to exclude prognostically significant coronary artery disease (CAD) in patients with left ventricular systolic dysfunction (LVSD). MATERIALS AND METHODS: A cohort of patients who underwent both X-ray angiography and CMR since 2006 was reviewed retrospectively. Records of those with European criteria for LVSD (left ventricular ejection fraction [LVEF] <50% or LV end-diastolic volume index [LVEDVI] ≥97 ml/m2) on CMR or transthoracic echo were analysed. The presence and extent of subendocardial late gadolinium enhancement (LGE) was recorded with the 17-segment model. The degree of coronary stenosis at X-ray angiography was assessed visually and significant disease defined as stenosis of the LMS ≥50%, or proximal left anterior descending ≥75%, or ≥70% in two main coronary vessels. RESULTS: One hundred and sixteen patients were included. The mean age was 64 years and 78% were male. The mean LVEF was 40%. The prevalence of prognostic CAD was 47%. The presence of subendocardial LGE detected prognostically significant CAD with a sensitivity of 100% (95% CI: 94-100%) with no false-negative results. CONCLUSIONS: The absence of subendocardial LGE on CMR reliably excludes prognostic CAD in patients with LVSD.

Oxygen use for chest pain in coronary care units across the UK.

AIM: To quantify the adherence to national guidance for the use of oxygen in patients presenting with chest pain to coronary care units (CCUs) across the UK. DESIGN: Prospective survey. METHODS: A total of 307 hospitals were contacted by telephone between August 2010 and October 2010. Of these, 48 had no CCUs, 10 units refused to take part and 18 hospitals were contacted on 2 occasions but were unable to provide the information due to paucity of time owing to heavy clinical workload. Overall 231 hospitals participated in the audit questionnaire. RESULTS: A total of 30% of the units used oxygen titrated to saturations in accordance with national guidelines. There was no difference between units that had on-site availability of percutaneous coronary intervention and those that did not. Those hospitals where there was a policy for routine oxygen prescription were as unlikely to comply with the guidelines on oxygen use as hospitals where oxygen was not routinely prescribed. CONCLUSION: Only one-third of CCUs in the UK reported adherence to guidelines with regards to oxygen delivery in patients presenting with chest pain. Despite this figure seeming rather low, this is consistent with practice through a range of specialties and guidelines. The evidence base for the oxygen guidance remains insecure. Additional research is required but in the meantime we recommend oxygen is prescribed according to current guidelines.

A model of nitrogen efficiency in contrasting grass-based dairy systems.

Nitrogen (N) efficiency is one of the key drivers of environmentally and economically sustainable agricultural production systems. An N balance model was developed, evaluated, and validated to assess N use efficiency and N surplus and to predict N losses from contrasting grass-based dairy production systems in Ireland. Data from a 5-yr study were used to evaluate and validate the model. Grass-based and high-concentrate production systems combined with 3 divergent strains of Holstein-Friesian (HF) dairy cows-high-production North American (HP), high-durability North American (HD), and New Zealand (NZ)-were evaluated. As concentrate input increased, N surplus per hectare increased and N use efficiency per hectare decreased (23 and 10%, respectively). When the N required to rear replacement animals to maintain the production system was considered, the N surplus of the HP genetic strain was greater (156 kg of N/cow) than that of the HD (140 kg of N/cow) or the NZ (128 kg of N/cow). The model estimated N leaching of 8.1mg of NO(3)-N/L, similar to that measured by others at the same site. The model creates awareness of methods and indicators available to assess the most suitable and environmentally sustainable grass based dairy production systems.

A comparison between Holstein-Friesian and Jersey dairy cows and their F(1) hybrid on milk fatty acid composition under grazing conditions.

The objective of this study was to investigate the effect of 2 breeds, Holstein and Jersey, and their F(1) hybrid (Jersey x Holstein) on milk fatty acid (FA) concentrations under grazing conditions, especially conjugated linoleic acid (CLA) and n-3 polyunsaturated fatty acids because of their importance to human health. Eighty-one cows (27 per breed grouping) were allocated a predominantly perennial ryegrass pasture. Samples were collected over 2 periods (June and July). Breed affected dry matter intake and milk production and composition. Holstein cows had the highest dry matter intake (18.4+/-0.40kg of DM/d) and milk production (21.1+/-0.53kg of DM/d). Holstein and Jersey x Holstein cows had similar 4% fat corrected milk, fat yield, and protein yield; with the exception of fat yield, these were all higher than for Jersey cows. Milk fat concentration was highest for Jersey cows and lowest for Holstein cows, with the hybrid cows intermediate. Total FA and linolenic acid intake (1.09+/-0.023 and 0.58+/-0.012 kg/d, respectively) were highest for Holstein cows. In terms of milk FA, Holstein cows had higher contents of C14:1, cis-9 C18:1 and linoleic acid. In turn, Jersey and Jersey x Holstein cows had higher content of C16:0. Milk concentrations of neither the cis-9,trans-11 isomer of CLA nor its precursor, vaccenic acid, were affected by breed. Nevertheless, large variation between individual animals within breed grouping was observed for CLA and estimated Delta(9)-desaturase activity. There was some evidence for a negative heterotic effect on milk concentration of CLA, with the F(1) hybrid cows having lower concentrations compared with the mid parent average. Plasma FA profile did not accurately reflect differences in milk FA composition. In conclusion, there was little evidence for either breed or beneficial heterotic effects on milk FA content with human health-promoting potential, though significant within-breed, interanimal variation was observed.

Extended lactations in a seasonal-calving pastoral system of production to modulate the effects of reproductive failure.

This study was conducted to determine whether extending the calving interval (CI) to 24 mo would be an alternative to culling and replacing cows that had failed to become pregnant. Forty-six nonpregnant lactating cows were assembled in November 2004 and assigned to receive either 3kg (low) or 6kg (high) of concentrate supplement and a basal diet of grass silage and maize silage over the winter period (13 wk). Cows returned to pasture in late March and received 1kg of concentrate/d until dry-off (milk yield <5 kg/d). Cumulative milk production was calculated from calving to the end of November 2004 (12-mo CI) and from the start of December 2004 until dry off in 2005 (extended lactation part of 24-mo CI). High winter feeding resulted in greater milk production over the winter confinement (20.0+/-0.3 vs. 17.8+/-0.3 kg/d for high and low winter feeding, respectively) and had a carryover effect during the remainder of the 24-mo CI period (5,177 vs. 4,686kg; SEM=173kg). At the end of the study, cows were ranked on cumulative milk solids and separated into 3 groups (R1, R2, and R3). During the 24-mo CI, milk yields were 7,287, 6,267, and 5,273kg (SEM=308kg) in yr 1, and 5,738, 4,836, and 4,266 (SEM=241kg) in yr 2 for R1, R2, and R3, respectively. Eighty-five percent of the cows became pregnant during the breeding season of yr 2, with a conception rate to first service of 52%. An economic analysis of different ranks with a 12-mo CI, a 24-mo CI, and an annualized herd effect, which compared an efficient spring calving system with 30% recycled cows in R1 and 10% recycled cows in R3, was carried out. Farm profit was reduced by 60% and 65% at a milk price of 22.3 euro-cents (c)/L with the corresponding values of 17% and 30% for a milk price of 30 c/L, respectively, when R1 and R3 systems were compared with an efficient spring milk (12-mo CI) production system. Within a spring system where 30% and 10% of R1 and R3 animals were subjected to extended lactations, the profit difference was reduced compared with an efficient spring system, The results indicated that lactations with a 24-mo CI may be a viable alternative to culling nonpregnant cows and be economically more suited to higher producing cows.

Short communication: the effect of dry period duration and dietary energy density in early lactation on the rennet gelation properties of milk.

This study was carried out to examine the effects of decreasing the dry period (DP) duration and altering the energy density of the diet during early lactation on the rheological characteristics of milk. Forty mature Holstein-Friesian cows were used in a completely randomized design with a 2 x 2 factorial arrangement of treatments. Cows were randomly assigned to 1 of 2 dry period treatments and 1 of 2 nutritional treatments. The DP treatments were continuous milking (CM) or an 8-wk standard dry period (SDP), and the nutritional treatments were a standard energy diet (SE) or a high energy diet (HE). Actual DP lengths were 6.3+/-1.7 d and 62.1+/-1.9 d for cows on the CM and SDP treatments, respectively. Milk samples were collected at 2, 6, and 10 wk postpartum. The concentrations of fat, protein, and lactose were determined in each sample. The rennet gelation properties were measured at 31 degrees C using dynamic low-amplitude strain oscillatory rheometry. The following parameters were obtained from the resultant elastic shear modulus (G'): gelation time, maximum curd firming rate, and gel strength. Reducing the DP duration from 62 to 6 d resulted in increases in milk protein concentration (31.8 vs. 34.7 g/kg), maximum curd firming rate (2.58 vs. 3.60 Pa/min), and gel strength (69.4 vs. 90.5Pa). Increasing the dietary energy density decreased percentage milk fat (43.1 vs. 37.7 g/kg) but otherwise had no effect. Gel strength was correlated with maximum curd firming rate (r=0.99), and both variables were correlated with milk protein concentration (r=0.71 and r=0.73, respectively). The results indicate that decreasing the duration of the DP increased milk protein concentration and improved the rennet gelation properties of milk, but that dietary energy density had little effect.

Effect of grain type and processing method on rumen fermentation and milk rumenic acid production.

It was hypothesized that differences in starch degradability account for observed differences in rumen vaccenic acid (t11-18:1) and milk rumenic acid (RA) concentrations. To test this hypothesis, starch degradability was varied through grain source and by processing. Eight Holstein cows in mid-lactation were assigned to two 4 × 4 Latin squares with four 21-day periods and four diets: dry rolled barley, ground barley, dry rolled corn and ground corn. Diets contained similar starch content and were supplemented with whole sunflower seed to provide similar total polyunsaturated fatty acid (PUFA) (18:2n-6 + 18:3n-3) contents. Forage/concentrate ratios of all diets were 42 : 58. Rumen, plasma and milk samples were collected in the third week of each period. In situ degradation rates (%/h) for rolled corn, ground corn, rolled barley and ground barley were 5.4, 8.9, 17.0 and 19.4, respectively, for dry matter (DM) and 6.3, 10.8, 25.3 and 43.8, respectively, for starch. DM intakes were greater for corn-based diets (CBD) than for barley-based diets (BBD) with no difference between rolled and ground diets. Daily minimum rumen pH was less (5.2 v. 5.5) and pH duration <5.8 (h/d) was greater (7.4 v. 4.3) for BBD than for CBD. Milk fat content and yield were less for BBD than for CBD with greater values observed for rolling compared with grinding. Variability in milk fat yield was strongly related (R2 = 0.55; P < 0.01) to total starch intake (45%) and milk c9t11-CLA (10%) and none of the t-18:1 isomers or CLA isomers that are typically associated with milk fat depression entered the model. The concentrations (%) of t10-18:1 and t11-18:1 were greater for BBD than for CBD in rumen contents (t10-18:1, 3.5 v. 1.3; t11-18:1, 3.2 v. 1.9), plasma (t10-18:1, 1.2 v. 0.2; t11-18:1, 0.97 v. 0.58) and milk (t10-18:1, 3.8 v. 1.0; t11-18:1, 2.6 v. 1.7) despite greater total PUFA intakes for CBD. Milk RA concentration was greater for BBD than for CBD (1.46 v. 0.89) but was not influenced by the method of grain processing. This study clearly demonstrated that the milk content and profile of t-18:1 and CLA isomers were more strongly influenced by the source of grain starch (barley > corn) than by the method of grain processing indicating that factors inherent in the source of starch were responsible for the observed differences and these factors could not be modified by the processing methods used in this study.

Fatty acid intake and milk fatty acid composition of Holstein dairy cows under different grazing strategies: herbage mass and daily herbage allowance.

The objective of this study was to investigate the effect of level of 1) pregrazing herbage mass (HM) and 2) level of daily herbage allowance (DHA) on the performance and fatty acid (FA) composition of milk from grazing dairy cows. Sixty-eight Holstein-Friesian dairy cows were allocated to either a high or low pregrazing HM (1,700 vs. 2,400 kg of DM/ha; >40 mm), and within HM treatment, cows were further allocated to either a high or low DHA (16 vs. 20 kg of DM/d per cow; >40 mm) in a 2 x 2 factorial design. Pregrazing HM did not affect dry matter intake (17.5 +/- 0.75 kg/d), milk production (22.1 +/- 0.99 kg/d), milk composition (milk fat, 3.88 +/- 0.114%; milk protein, 3.28 +/- 0.051%), body weight (525 +/- 16 kg), or body condition score (2.65 +/- 0.064). Increasing DHA increased dry matter intake (+1.5 kg/d) but did not affect any other variable measured. Cows grazing the low HM or high DHA had a higher daily intake of total FA (+0.12 and +0.09 kg/d, respectively, for the low HM and high DHA), alpha-linolenic acid (LNA; +0.08 and +0.05 kg/d, respectively, for the low HM and high DHA), and linoleic acid (+0.01 for both the low HM and high DHA) compared with either the high HM or low DHA. Milk conjugated linoleic acid (cis-9, trans-11 isomer) was not affected by treatment (13.0 +/- 0.77 g/kg of total FA); however, large variation was recorded between individual animals (range from 5.9 to 20.6 g/kg of total FA). Milk concentrations of LNA were higher for animals offered the low HM (5.3 g/kg of total FA), but across treatments, milk concentrations of LNA were low (4.9 +/- 0.33 g/kg of total FA). The present study indicates that changes in HM and DHA do not have a great effect on the milk FA composition of grazing dairy cows. Further enhancement of the beneficial FA content in milk purely from changes in grazing strategy may be difficult when pasture quality is already high.

Pleiotropic effects of negative energy balance in the postpartum dairy cow on splenic gene expression: repercussions for innate and adaptive immunity.

Increased energy demands to support lactation, coupled with lowered feed intake capacity results in negative energy balance (NEB) and is typically characterized by extensive mobilization of body energy reserves in the early postpartum dairy cow. The catabolism of stored lipid leads to an increase in the systemic concentrations of nonesterified fatty acids (NEFA) and beta-hydroxy butyrate (BHB). Oxidation of NEFA in the liver result in the increased production of reactive oxygen species and the onset of oxidative stress and can lead to disruption of normal metabolism and physiology. The immune system is depressed in the peripartum period and early lactation and dairy cows are therefore more vulnerable to bacterial infections causing mastitis and or endometritis at this time. A bovine Affymetrix oligonucleotide array was used to determine global gene expression in the spleen of dairy cows in the early postpartum period. Spleen tissue was removed post mortem from five severe NEB (SNEB) and five medium NEB (MNEB) cows 15 days postpartum. SNEB increased systemic concentrations of NEFA and BHB, and white blood cell and lymphocyte numbers were decreased in SNEB animals. A total of 545 genes were altered by SNEB. Network analysis using Ingenuity Pathway Analysis revealed that SNEB was associated with NRF2-mediated oxidative stress, mitochondrial dysfunction, endoplasmic reticulum stress, natural killer cell signaling, p53 signaling, downregulation of IL-15, BCL-2, and IFN-gamma; upregulation of BAX and CHOP and increased apoptosis with a potential negative impact on innate and adaptive immunity.

Grazing cows are more efficient than zero-grazed and grass silage-fed cows in milk rumenic acid production.

Six rumen-cannulated Holstein cows in early lactation were assigned to 3 treatments: grazing (G), zero-grazing (ZG), and grass silage (GS) harvested from the same perennial rye grass sward in a 3 x 3 Latin square design with three 21-d periods. The objectives of this study were to investigate the underlying mechanisms for the reported elevation in milk rumenic acid (RA) concentration associated with G compared with ZG and GS, and to identify the important variables contributing to the milk RA response. Grazing animals were offered 20 kg of dry matter/cow per day; indoor animals were offered ad libitum grass or silage. A concentrate at a rate of 3 kg/d was also offered to all cows. Rumen, plasma, and milk samples were collected in the third week of each period. Data were analyzed by the MIXED procedure of SAS. Dry matter intakes were less for GS with no difference between G and ZG. Milk yield was greater for G than for ZG or GS. Milk fat and protein contents were less for GS with no difference between G and ZG. The combined intake (g/d) of linoleic and linolenic (18:3n-3) acids was different across the treatments (G: 433; ZG: 327; and GS: 164). Rumen pH was less for G with no difference between ZG and GS. Concentrations of volatile fatty acids and ammonia nitrogen in rumens were not different across the treatments. Wet rumen fill was less for G with no difference between ZG and GS. Vaccenic acid concentrations were different across the treatments in rumen (G: 12.30%, ZG: 9.31%, and GS: 4.21%); plasma (G: 2.18%, ZG: 1.47%, and GS: 0.66%) and milk (G: 4.73%, ZG: 3.49%, and GS: 0.99%). Milk RA concentrations were greater for G (2.07%) than for ZG (1.38%) and GS (0.54%). Milk desaturase index based on the ratio cis-9-14:1/14:0 was not different across the treatments. Milk RA yield per 100 g of linoleic acid and linolenic acid intake (efficiency) was 2.23, 1.50, and 0.62 g in G, ZG, and GS, respectively, suggesting that G cows were more efficient than ZG and GS cows in milk RA production. Stepwise regression analysis of a group of variables revealed that plasma vaccenic acid accounted for 95% of the variation in milk RA production. Milk desaturase index did not enter into the model. Overall findings suggest that substrate intake influenced milk RA production but it was not the only factor involved. There were differences in efficiency of milk RA production, which appears to depend on the factors regulating ruminal vaccenic acid production and its supply to the mammary tissue.

Responses of North American and New Zealand strains of Holstein-Friesian dairy cattle to homeostatic challenges during early and mid-lactation.

This study investigated the physiological basis of differences in nutrient partitioning between the North American (NA) and New Zealand (NZ) strains of Holstein-Friesian cattle by determining the responses to homeostatic challenges at two stages of lactation. Glucose tolerance tests, epinephrine challenges and insulin challenges were carried out on consecutive days commencing on day 32 ± 0.48 (mean ± s.e.) of lactation (T1) and again commencing on day 137 ± 2.44 of lactation (T2). The insulin and non-esterified fatty acid (NEFA) responses to glucose infusion did not differ between the strains. The NZ strain had a greater clearance rate (CR) of glucose (2.04% v. 1.66%/min) and tended to have a shorter (34.4 v. 41.1 min) glucose half-life (t½) at T2 when infused with glucose. The NA cows had a greater glucose response to epinephrine infusion across T1 and T2, and tended to have a greater insulin response to epinephrine infusion. Plasma NEFA concentration declined to similar nadir concentrations for both strains at T1 in response to insulin, though from a higher basal concentration in NA cows, resulting in a greater (-2.29 v. -1.38) NEFA area under the response curve for NA cows. Glucose response to insulin varied with time, tending to be greater for NA at T1, but tending to be lower for NA at T2. The results indicated that NA cows had a greater glycogenolytic response to epinephrine, but both strains had similar lipolytic responses. The results also imply that higher basal circulating NEFA concentrations in the NA strain in early lactation were not due to diminished adipose tissue responsiveness to insulin. There were indications that glucose CR was greater in NZ cows in mid-lactation, and may form the basis of increased body tissue accretion during mid- to late-lactation in this strain.

Negative energy balance in dairy cows is associated with specific changes in IGF-binding protein expression in the oviduct.

Negative energy balance (NEB) during early lactation in dairy cows leads to an altered metabolic state that has major effects on the production of IGF family members. Low IGF-I concentrations are associated with poor fertility and therefore we aimed to determine whether NEB exerts a direct effect on IGF expression in the postpartum oviduct. Multiparous Holstein cows were allocated to two treatments (each n=6) designed using differential feeding and milking regimes to produce either mild NEB (MNEB) or severe NEB (SNEB). Animals were slaughtered in week 2 of lactation when divergent metabolic profiles were evident. Oviducts were collected for RNA analysis by real-time RT-PCR and in situ hybridisation. Quantitative measures in oviduct gene expression were obtained for all members of the IGF family (IGF-I/II, IGF-binding proteins (IGFBP) 1-6 and receptors for IGF types 1 and 2), insulin A/B, GH, glucocorticoid and oestrogen alpha/beta. Expression of IGFBP-2 and IGFBP-6 (both of which have a high affinity for IGF-II) was decreased in SNEB relative to MNEB (P<0.05). No other gene was altered by NEB, but IGF-II, IGFBP-3, IGFBP-5 and IGFBP-6 all showed differential expression in different regions of the oviduct. These results indicate that, in addition to low circulating IGF-I after calving, NEB may also influence IGF availability in the oviduct indirectly through changes in specific IGFBP expression. It is possible that the predicted increased signalling by IGF-II may perturb embryo development, contributing to the high rates of embryonic mortality in dairy cows.

A comparison of energy balance and metabolic profiles of the New Zealand and North American strains of Holstein Friesian dairy cow.

The milk production, energy balance (EB), endocrine and metabolite profiles of 10 New Zealand Holstein Friesian (NZ) cows and 10 North American Holstein Friesian (NA) cows were compared. The NA cows had greater peak milk yields and total lactation milk yields (7387 v. 6208 kg; s.e.d. = 359), lower milk fat and similar protein concentrations compared with the NZ cows. Body weight (BW) was greater for NA cows compared with NZ cows throughout lactation (596 v. 544 kg; s.e.d. = 15.5), while body condition score (BCS) tended to be lower. The NA strain tended to have greater dry matter intake (DMI) (17.2 v. 15.7 kg/day; s.e.d. = 0.78) for week 1 to 20 of lactation, though DMI as a proportion of metabolic BW was similar for both strains. No differences were observed between the strains in the timing and magnitude of the EB nadir, interval to neutral EB, or mean daily EB for week 1 to 20 of lactation. Plasma concentrations of glucose and insulin were greater for NA cows during the transition period (day 14 pre partum to day 28 post partum). Plasma IGF-I concentrations were similar for the strains at this time, but NZ cows had greater plasma IGF-I concentration from day 29 to day 100 of lactation, despite similar calculated EB. In conclusion, the results of this study do not support the premise that the NZ strain has a more favourable metabolic status during the transition period. The results, however, indicate that NZ cows begin to partition nutrients towards body reserves during mid-lactation, whereas NA cows continue to partition nutrients to milk production.

Cheese manufacture with milk with elevated conjugated linoleic acid levels caused by dietary manipulation.

The objective of this study was to assess the effect of dietary supplementation of cows on pasture with sunflower oil for conjugated linoleic acid (cis-9, trans-11 CLA) enrichment of milk, for the production of CLA-enriched cheese. A group of 40 autumn-calving dairy cows were assigned to either a control group (indoor feeding on grass silage ad libitum and 6 kg/d of a typical indoor concentrate) or an experimental group (on pasture, being fed 6 kg of a supplement containing 100 g/kg of sunflower oil per d). These diets were fed for 16 d, during which time milk was collected for pilot-scale hard cheese manufacture. The pasture-based diet with sunflower oil resulted in a significant effect on the milk fatty acid CLA content. The concentration of cis-9, trans-11 CLA in the milk produced from cows on this diet increased to 2.22 g/100 g of fatty acid methyl esters (FAME) after 14 d, compared with 0.46 g/100 g of FAME in milk produced on the control indoor diet. The content of cis-9, trans-11 CLA in the cheese manufactured from the indoor control milk was 0.78 g/100 g of FAME and that from the pasture-based sunflower oil milk was 1.93 g/100 g of FAME. The cheese was assessed during the ripening period and CLA concentrations were stable throughout the 6 mo of ripening. Other cheese variables (microbiology, composition, flavor, free AA) were monitored during the ripening period, and the cheese with the elevated CLA concentrations compared favorably with the control cheese. Thus, a pasture-based diet supplemented with an oil source rich in linoleic acid resulted in an enhanced CLA content of bovine milk fat, compared with an indoor grass silage-based diet.

Effect of negative energy balance on the insulin-like growth factor system in pre-recruitment ovarian follicles of post partum dairy cows.

Post partum negative energy balance (NEB) in dairy cattle is associated with a delayed return to ovarian cyclicity and reduced fertility. This study compared the IGF system of pre-recruitment ovarian follicles between cows in mild (n = 6) or severe (n = 6) NEB during early lactation. Ovaries were collected in the second week post partum, when circulating concentrations of IGF-I and glucose were lower (P < 0.01) in severe NEB cows. mRNA expression for IGF-II, type 1 IGF receptor (IGF-1R) and IGF-binding proteins (IGFBP)-1 to IGFBP-6 was determined by in situ hybridisation in individual follicles using radiolabelled oligonucleotide probes. Follicles were classified as very small (1-2.5 mm) or small (2.5-5 mm) and healthy or atretic. Relative mRNA concentrations were measured as optical density (OD) units using image analysis. Thecal IGF-II mRNA expression was highest in very small, healthy follicles (P < 0.05). Granulosa cell IGFBP-2 was the only component to change with EB status, with higher mRNA expression in mild compared with severe NEB cows (P < 0.05). IGFBP-1 and IGFBP-3 mRNA expression were undetectable. IGF-1R, IGFBP-4 and IGFBP-5 mRNA expression were not significantly altered by follicle size or health, but IGFBP-5 tended to increase in atretic follicles. The pattern of IGFBP-6 mRNA expression in theca paralleled that of IGF-II mRNA, with higher (P < 0.05) levels in healthy, very small follicles. In conclusion, the reduced expression of IGFBP-2 mRNA in severe NEB cows may alter the bioavailability of circulating IGF-I and locally produced IGF-II to modulate the pre-recruitment stages of follicles required to maintain normal post partum ovarian cyclicity.

Relationships among milk production, energy balance, plasma analytes, and reproduction in Holstein-Friesian cows.

Associations were examined between components and indicators of early lactation energy balance (EB) and measures of fertility in Holstein cows. Milk production, dry matter intake (DMI), body condition score (BCS), and endocrine and metabolite data from 96 cows were analyzed using multivariate logistic regression and survival analysis. Fertility variables investigated were interval to commencement of luteal activity (C-LA), calving to conception interval (CCI), and conception rate to first service (CON1). Mean daily EB, milk protein content, and DMI during the first 28 d in milk were associated positively with CON1. Cows having poorer BCS (< or =2.25) at first service had a lower CON1. Positive associations were identified among EB, milk protein content, DMI, and the likelihood of a shorter interval to C-LA. Cows having greater DMI and a more positive EB had an increased likelihood of a shorter CCI, whereas a lower nadir BCS was associated with an increased likelihood of a longer CCI. Milk yield was not associated with any of the fertility variables investigated. A greater plasma concentration of insulin-like growth factor I (IGF-I) during the first 2 wk of lactation was associated with a greater CON1 and an increased likelihood of a shorter interval to C-LA. In conclusion, we identified DMI as the principal component of EB influencing subsequent fertility. Furthermore, results indicate that milk protein content and plasma IGF-I concentration in early lactation may be useful indicators of reproductive efficiency.

Comparative evaluation of alternative forages to grass silage in the diet of early lactation dairy cows.

Fifty-six autumn-calving Holstein-Friesian cows, blocked on the basis of days in milk (27.6 +/- 10.65 d), lactation number (3.1 +/- 2.21), and preexperimental milk yield (28.4 +/- 6.69 kg) were used to examine the effects of replacing 330 g/kg of dry matter (DM) of first-cut perennial ryegrass silage with either fermented whole-crop wheat (WCW), urea-treated processed WCW, or corn silage on subsequent feed intake, milk production, and efficiency of nitrogen utilization. The DM (g/kg), crude protein (CP, g/kg of DM) and in vitro DM digestibility (g/kg) of the forages were 204, 179, and 762 for grass silage; 389, 90, and 711 for fermented WCW; 795, 141, and 768 for urea-treated processed WCW; and 346, 93, and 783 for corn silage, respectively. Four forage treatments were evaluated as follows: 1) grass silage as the sole forage (GS); 2) a mixture of grass silage and fermented WCW silage, (F-WCW); 3) a mixture of grass silage and urea-treated processed WCW, (UP-WCW); and 4) a mixture of grass silage and corn silage (CS). In all cases, the alternative forages comprised 67% of the forage mix on a DM basis. Isonitrogenous diets were formulated by offering all cows 8 kg of concentrate as fed, formulated to different CP concentrations. Cows were offered these diets from 28 to 104 d in milk. Total DM intake and milk yield were greater on UP-WCW (20.0 and 30.2 kg/d) and CS (18.3 and 33.2 kg/d) than on GS (13.5 and 26.5 kg/d). Although DM intake was greater on F-WCW (17.1 kg/d) than on GS, milk yield was not significantly greater (+2.7 kg/d). Milk protein concentration was greater on F-WCW (30.5 g/kg), UP-WCW (31.3 g/kg), and CS (30.7 g/kg) than on GS (28.5 g/kg). However, there was no difference between treatments in milk fat or lactose concentrations. Body weight change was greater for cows offered GS (-0.27 kg/d) than for those offered UP-WCW (-0.01 kg/d) and CS (+0.05 kg/d) but not compared with those offered F-WCW (-0.06 kg/d). There was no effect of treatment on plasma glucose, nonesterified fatty acids, beta-hydroxybutyrate, urea, or total protein at d 64 +/- 17.4 and d 92 +/- 17.4 postpartum. Efficiency of N utilization was greatest for CS with 0.36 of N intake being recovered in milk compared with 0.28, 0.32, and 0.26 for GS, F-WCW, and UP-WCW, respectively. There was no effect of treatment on milk urea N concentration or the urinary allantoin N to creatinine N ratio. The results of this experiment indicate that corn silage is a more suitable supplementary forage to grass silage than fermented or urea-treated processed WCW, with advantages realized in milk production and more efficient N utilization.

Anti-beta2GPI-antibody-induced endothelial cell gene expression profiling reveals induction of novel pro-inflammatory genes potentially involved in primary antiphospholipid syndrome.

OBJECTIVE: To determine the effects of primary antiphospholipid syndrome (PAPS)-derived anti-beta(2)GPI antibodies on gene expression in human umbilical vein endothelial cells (HUVEC) by gene profiling using microarrays. METHODS: Anti-beta(2)GPI antibodies purified from sera of patients with PAPS or control IgG isolated from normal subjects were incubated with HUVEC for 4 h before isolation of RNA and processing for hybridisation to Affymetrix Human Genome U133A-2.0 arrays. Data were analysed using a combination of the MAS 5.0 (Affymetrix) and GeneSpring (Agilent) software programmes. For selected genes microarray data were confirmed by real-time PCR analysis or at the protein level by ELISA. RESULTS: A total of 101 genes were found to be upregulated and 14 genes were downregulated twofold or more in response to anti-beta(2)GPI antibodies. A number of novel genes not previously associated with APS were induced, including chemokines CCL20, CXCL3, CX3CL1, CXCL5, CXCL2 and CXCL1, the receptors Tenascin C, OLR1, IL-18 receptor 1, and growth factors CSF2, CSF3 IL-6, IL1beta and FGF18. The majority of downregulated genes were transcription factors/signalling molecules including ID2. Quantitative real-time RT-PCR analysis confirmed the microarray results for selected genes (CSF3, CX3CL1, FGF18, ID2, SOD2, Tenascin C). CONCLUSIONS: This study reveals a complex gene expression response in HUVEC to anti-beta(2)GPI antibodies with multiple chemokines, pro-inflammatory cytokines, pro-thrombotic and pro-adhesive genes regulated by these antibodies in vitro. Some of these newly identified anti-beta(2)GPI antibody-regulated genes could contribute to the vasculopathy associated with this disease.

Embryo survival in dairy cows managed under pastoral conditions.

Efficient pasture-based milk production systems require a compact calving pattern aligned to the onset of the grazing season, a 365-day calving interval and low culling rates for infertility. Achievement of these targets requires high herd reproductive performance. While high genetic merit Holstein cows produce more milk in grass-based systems their fertility is compromised. Management of the modern high genetic merit Holstein dairy cow presents a major challenge in pasture-based systems of production. It appears that the extent of early embryo loss is greater (up to 20% points greater) in the modern high-producing dairy cow and that a much higher proportion of the embryos die before day 7 following insemination in contrast to heifers and lower yielding cows. About 7-8% of pregnancies are lost between days 30 and 90 of gestation with no evidence that loss rate is related to cow genetic merit, parity or level of production. Systemic concentrations of progesterone during both the cycle preceding and following insemination affect embryo survival rate with evidence that too low or indeed too high a concentration of progesterone been negatively associated with embryo survival rate. Peripheral concentrations of both progesterone and oestradiol are lowered by increased plane of feed intake due to increased metabolic clearance rate of the steroids, which is related to liver blood flow. It appears that high producing dairy cows have an increased risk of embryo death as a result of lowered peripheral concentrations of progesterone as a consequence of increased hepatic metabolism of progesterone. Uterine expression of mRNA for progesterone receptor, oestradiol receptor and retinol binding protein mRNA appears to be sensitive to changes in peripheral concentrations of progesterone during the first week after AI. It would appear that energy balance and dry matter intake during the 4 weeks, immediately after calving are critically important in determining conception rate when cows are inseminated at 70-100 days post-calving. Concentrate supplementation of cows at pasture during the breeding period has minimal affects on conception rates though sudden reduction in dietary intake should be avoided. For pasture-based systems of milk production more balanced breeding strategies, with greater emphasis on fertility and feed intake must be developed.