The acute effects of rumen pulse-dosing of hydrogen acceptors during methane inhibition with nitrate or 3-nitrooxypropanol in dairy cows.

Dietary methane (CH4) mitigation is in some cases associated with an increased hydrogen (H2) emission. The objective of the present study was to investigate the acute and short-term effects of acceptors for H2 (fumaric acid, acrylic acid or phloroglucinol) supplemented via pulse-dosing to dairy cows fed CH4 mitigating diets (using nitrate or 3-nitrooxypropanol), on gas exchange, rumen gas and VFA composition. For this purpose, 2 individual 4 × 4 Latin square experiments were conducted with 4 periods of 3 d (nitrate supplementation) and 7 d (3-nitrooxypropanol supplementation), respectively. In each study, 4 rumen cannulated Danish Holstein cows were used. Each additive for CH4 mitigation was included in the ad libitum fed diet within the 2 experiments, to which the cows were adapted for at least 14 d. Acceptors for H2 were administered twice daily in equal portions through the rumen fistula immediately after feeding of the individual cow. In Exp. 1 (nitrate), the treatments were CON-1 (no H2-acceptor), FUM-1 (fumaric acid), ACR-1 (acrylic acid) and FUM+ACR-1 (50% FUM-1 + 50% ACR-1). In Exp. 2 (3-nitrooxypropanol), the 3 treatments, CON-2, FUM-2, and ACR-2, were similar to CON-1, FUM-1 and ACR-1 treatments, however the fourth treatment was PHL-2 (phloroglucinol). Gas exchanges were measured in respiration chambers, while samples of rumen liquid and headspace gas were taken in time series relative to feeding and dosing on specific days. Headspace gas was analyzed for gas composition and rumen liquid was analyzed for volatile fatty acid composition and dissolved gas concentrations. Headspace gas composition and dissolved gas concentration were only measured in Exp. 2. Dry matter intake was reduced upon acrylic acid supplementation. There were no significant effects of any treatments in any experiments on H2 emission, except for a decrease in hourly H2 emission rate (g/h) at 1 h after feeding in both experiments. In Exp. 2, H2 headspace proportions increased by ACR-2 supplementation, whereas dissolved concentrations were unaffected. In Exp. 1, cows on ACR-1 increased propionate proportion at 1 h after feeding. In Exp. 2, both FUM-2 and ACR-2 increased rumen propionate proportion in the hours after feeding and dosing. There was no effect on rumen acetate for cows on PHL-2. There was a strong positive correlation between rumen dissolved CH4 and headspace CH4 (r = 0.84), whereas the equivalent correlation was weaker for H2 (r = 0.41). For the relationship between dissolved concentrations and emissions of CH4 and of H2, there was a moderate positive correlation for CH4 (r = 0.54), whereas it was weak for H2 (r = 0.28) with zero slope. In conclusion, the results suggested that fumaric acid and acrylic acid to some extent was reduced to propionate without associative effects on measures for H2 redirection. Furthermore, phloroglucinol seemed not to be metabolized in the rumen in the present study, as no effects on rumen acetate or measures of H2 were observed. Changes in H2 headspace and emission may be a poor proxy for actual changes in the rumen fluid concentration of H2.

Gas exchange, rumen hydrogen sinks, and nutrient digestibility and metabolism in lactating dairy cows fed 3-nitrooxypropanol and cracked rapeseed.

Fat in the form of cracked rapeseed and 3-nitrooxypropanol (3-NOP, market as Bovaer) were fed alone or in combination to 4 Danish Holstein multicannulated dairy cows, with the objective to investigate effects on gas exchange, dry matter intake (DMI), nutrient digestion, and nutrient metabolism. The study design was a 4 × 4 Latin square with a 2 × 2 factorial treatment arrangement with 2 levels of fat supplementation; 33 g of crude fat per kg of dry matter (DM) or 64 g of crude fat per kg of DM for low and high fat diets, respectively, and 2 levels of 3-NOP; 0 mg/kg DM or 80 mg/kg DM. In total, 4 diets were formulated: low fat (LF), high fat (HF), 3-NOP and low fat (3LF), and 3-NOP and high fat (3HF). Cows were fed ad libitum and milked twice daily. The adaptation period lasted 11 d, followed by 5 d with 12 diurnal sampling times of digesta and ruminal fluid. Thereafter, gas exchange was measured for 5 d in respiration chambers. Chromic oxide and titanium dioxide were used as external flow markers to determine intestinal nutrient flow. No interactions between fat supplementation and 3-NOP were observed for methane yield (g/kg DM), total-tract digestibility of nutrients or total volatile fatty acid (VFA) concentration in the rumen. Methane yield (g/kg DMI) was decreased by 24% when cows were fed 3-NOP. In addition, 3-NOP increased carbon dioxide and hydrogen yield (g/kg DM) by 6% and 3,500%, respectively. However, carbon dioxide production was decreased when expressed on a daily basis. Fat supplementation did not affect methane yield but tended to reduce methane in percent of gross energy intake. A decrease (11%) in DMI was observed, when cows were fed 3-NOP. Likely, the lower DMI mediated a lower passage rate causing the tendency to higher rumen and total-tract neutral detergent fiber digestibility, when the cows were fed 3-NOP. Total VFA concentrations in the rumen were negatively affected both by 3-NOP and fat supplementation. Furthermore, 3-NOP caused a shift in the VFA fermentation profile, with decreased acetate proportion and increased butyrate proportion, whereas propionate proportion was unaffected. Increased concentrations of the alcohols methanol, ethanol, propanol, butanol, and 2-butanol were observed in the ruminal fluid when cows were fed 3-NOP. These changes in rumen metabolites indicate partial re-direction of hydrogen into other hydrogen sinks, when methanogenesis is inhibited by 3-NOP. In conclusion, fat supplementation did not reduce methane yield, whereas 3-NOP reduced methane yield, irrespective of fat level. However, the concentration of 3-NOP and diet composition and resulting desired mitigation effect must be considered before implementation. The observed reduction in DMI with 80 mg 3-NOP/kg DM was intriguing and may indicate that a lower dose should be applied in a Northern European context; however, the mechanism behind needs further investigation.

Childhood adversity and cardiovascular disease in early adulthood: a Danish cohort study.

AIMS: To examine the effect of childhood adversity on the development of cardiovascular disease (CVD) between ages 16 and 38, specifically focusing on ischaemic heart disease and cerebrovascular disease. METHODS AND RESULTS: Register data on all children born in Denmark between 1 January 1980 and 31 December 2001, who were alive and resident in Denmark without a diagnosis of CVD or congenital heart disease until age 16 were used, totalling 1 263 013 individuals. Cox proportional hazards and Aalen additive hazards models were used to estimate adjusted hazard ratios (HRs) and adjusted hazard differences of CVD from ages 16 to 38 in five trajectory groups of adversity experienced between ages 0 and 15. In total, 4118 individuals developed CVD between their 16th birthday and 31 December 2018. Compared with those who experienced low levels of adversity, those who experienced severe somatic illness and death in the family (men: adjusted HR: 1.6, 95% confidence interval: 1.4-1.8, women: 1.4, 1.2-1.6) and those who experienced very high rates of adversity across childhood and adolescence (men: 1.6, 1.3-2.0, women: 1.6, 1.3-2.0) had a higher risk of developing CVD, corresponding to 10-18 extra cases of CVD per 100 000 person-years in these groups. CONCLUSIONS: Individuals who have been exposed to childhood adversity are at higher risk of developing CVD in young adulthood compared to individuals with low adversity exposure. These findings suggest that interventions targeting the social origins of adversity and providing support for affected families may have long-term cardio-protective effects.

Effects of feeding level and milking frequency on behavior of dairy cows before dry-off.

Drying-off may challenge the welfare of especially high-yielding dairy cows. In this study, 119 loose-housed Holstein cows yielding ≥20 kg milk/d were enrolled in a 2 × 2 factorial design aiming to investigate effects of changes in diet energy density and daily milking frequency on behavior. The last 7 d before the dry-off day, cows were milked either twice or once daily, and were fed either a lactation diet or the same diet diluted with 30% barley straw, both offered in individual bins for ad libitum intake. All cows were fitted with sensors to record lying time and activity, and data from 109 of these cows were used together with behavioral observations obtained from video recordings of 52 of the cows. Data from activity sensors and video recordings were obtained during 24 h on d -6, -3, and -1 relative to the dry-off day (i.e., the day of the last milking). Across all days of observation, cows milked once daily spent more time feeding (149 vs. 130 min/d) than cows milked twice daily. Cows on the reduced diet and milked twice daily had a shorter lying time compared with cows on the normal diet and milked twice (759 vs. 837 min/d), whereas lying times of cows on the remaining 2 treatments were intermediate. Among cows on the lactation diet, reduced milking frequency increased time spent perching (from 11.1 to 28.7 min/d). Cows fed the energy-reduced diet spent more time feeding (154 vs. 124 min/d), showed more attempts to feed from unassigned feed bins (31.7 vs. 15.4 attempts daily), and spent less time using a mechanical brush (6.5 vs. 9.2 min/d) than cows fed the lactation diet. These results show that several aspects of cows' behavior, including main activities such as lying and feeding, but also behaviors of low resilience such as brush use, and to some extent more subtle and complex behaviors such as perching and attempts to feed from unassigned feed bins, are sensitive to management changes typically applied during the days before dry-off. The behavioral effects of the reduced feed energy level support earlier findings suggesting that qualitative feed restriction renders cows hungry. The effects of the reduced milking frequency on behavior were generally less pronounced, but 2 noteworthy interactions between milking frequency and diet were seen. First, cows milked twice daily and fed the energy-reduced diet spent less time lying, which is possibly related to increased energy demand and hunger. Second, cows milked once daily and fed the lactation diet spent more time perching, which may be related to udder discomfort. However, these more complex findings warrant further study. Taken together, the results of this study show that a dry-off procedure involving reduced energy supply induces behavioral changes indicating a higher degree of compromised welfare compared with reduced milking frequency.

Effects of feed energy density, daily milking frequency, and a single injection of cabergoline on behavior and welfare in dairy cows at dry-off.

Drying off dairy cows may challenge animal welfare due to high milk yields. A total of 111 loose-housed Holstein cows yielding >15 kg/d of milk were included in a 2 × 2 × 2 factorial design during dry-off to investigate the effects of reduced feeding level (normal vs. reduced energy density), reduced milking frequency (twice vs. once daily), and administration of a dopamine agonist (saline i.m. injection vs. cabergoline i.m. injection) on behavior in the home pen. During the 7 d before dry-off, cows were fed and milked according to 1 of the 4 feeding level and milking frequency combinations. Within 3 h after the last milking, cows were injected i.m. with 5 mL of either saline or a dopamine agonist (5.6 mg of cabergoline; Velactis, Ceva Santé Animale, Libourne, France; labeled for use only with abrupt dry-off, i.e., no preceding reduction in feeding level or milking frequency before last milking). Cows' behavior during d -1, 0, and +1 relative to the last milking was recorded via video and leg-attached sensors. Cows on the reduced energy density diet spent more time feeding and showed more attempts to feed from other cows' bins on d -1. Throughout the period of observations, cows on the reduced diet spent a lower percentage of lying time with their head raised, a higher percentage of lying time with their legs bent, and less time standing in a vigilant posture than did cows on the normal lactation diet. Reducing the daily milking frequency from 2 to 1 did not result in any clear behavioral signs of discomfort. On d 0, cows injected with cabergoline lay down longer but had their head raised for a shorter percentage of time while lying, compared with cows injected with saline. Cows injected with cabergoline also spent less time feeding than cows injected with saline on d 0, and reduced the time spent drinking from d -1 to d 0. Finally, fewer cabergoline-injected cows used the brush for self-grooming, and, among cows that did use the brush, the cows injected with cabergoline reduced the time spent using the brush from d -1 to d 0. In conclusion, cows injected with cabergoline showed several behavioral changes compared with control cows injected with saline. The behavioral changes shown by cows injected with cabergoline may be indicative of malaise during the first 24 h after injection, raising concern for animal welfare. No behavioral evidence for reduced udder pain in cows injected with cabergoline compared with control cows injected with saline was found. Drying off by reducing the energy density of the diet caused behavioral changes indicative of hunger before dry-off, whereas reducing the milking frequency had no clear effects on behavior.

Effect of nitrate supplementation, dietary protein supply, and genetic yield index on performance, methane emission, and nitrogen efficiency in dairy cows.

The objective was to investigate the effect of nonprotein nitrogen source, dietary protein supply, and genetic yield index on methane emission, N metabolism, and ruminal fermentation in dairy cows. Forty-eight Danish Holstein dairy cows (24 primiparous cows and 24 multiparous cows) were used in a 6 × 4 incomplete Latin square design with 4 periods of 21-d duration. Cows were fed ad libitum with the following 6 experimental diets: diets with low, medium, or high rumen degradable protein (RDP):rumen undegradable protein (RUP) ratio (manipulated by changing the proportion of corn meal, corn gluten meal, and corn gluten feed) combined with either urea or nitrate (10 g NO3-/kg of dry matter) as nonprotein nitrogen source. Samples of ruminal fluid and feces were collected from multiparous cows, and total-tract nutrient digestibility was estimated using TiO2 as flow marker. Milk samples were collected from all 48 cows. Gas emission (CH4, CO2, and H2) was measured by 4 GreenFeed units. We observed no significant interaction between dietary RDP:RUP ratio and nitrate supplementation, and between nitrate supplementation and genetic yield index on CH4 emission (production, yield, intensity). As dietary RDP:RUP ratio increased, intake of crude protein, RDP, and neutral detergent fiber and total-tract digestibility of crude protein linearly increased, and RUP intake linearly decreased. Yield of milk, energy-corrected milk, and milk protein and lactose linearly decreased, whereas milk fat and milk urea nitrogen concentrations linearly increased as dietary RDP:RUP ratio increased. The increase in dietary RDP:RUP ratio resulted in a linear increase in the excretion of total purine derivatives and N in urine, but a linear decrease in N efficiency (milk N in % of N intake). Nitrate supplementation reduced dry matter intake (DMI) and increased total-tract organic matter digestibility compared with urea supplementation. Nitrate supplementation resulted in a greater reduction in DMI and daily CH4 production and a greater increase in daily H2 production in multiparous cows compared with primiparous cows. Nitrate supplementation also showed a greater reduction in milk protein and lactose yield in multiparous cows than in primiparous cows. Milk protein and lactose concentrations were lower for cows receiving nitrate diets compared with cows receiving urea diets. Nitrate supplementation reduced urinary purine derivatives excretion from the rumen, whereas N efficiency tended to increase. Nitrate supplementation reduced proportion of acetate and propionate in ruminal volatile fatty acids. In conclusion, no interaction was observed between dietary RDP:RUP ratio and nitrate supplementation, and no interaction between nitrate supplementation and genetic yield index on CH4 emission (production, yield, intensity) was noted. Nitrate supplementation resulted in a greater reduction in DMI and CH4 production, and a greater increase in H2 production in multiparous cows than in primiparous cows. As the dietary RDP:RUP ratio increased, CH4 emission was unaffected and RDP intake increased, but RUP intake and milk yield decreased. Genetic yield index did not affect CH4 production, yield, or intensity.

Effects of feeding level, milking frequency, and single injection of cabergoline on blood metabolites, hormones, and minerals around dry-off in dairy cows.

This study aimed to investigate the effect of the different dry-off strategies based on reducing feeding level (normal vs. reduced energy density), reducing milking frequency (twice vs. once daily), and administration of a dopamine agonist after last milking (i.e. saline vs. cabergoline injection) on blood metabolites, hormones, and minerals around dry-off. In this experiment, 119 Holstein dairy cows were used in a 2 × 2 × 2 factorial arrangement. In the last week before dry-off, cows were allocated to 1 of the 4 possible dry-off strategies based on feeding level and milking frequency. Within 3 h after last milking, cows were injected with either saline or a D2 dopamine agonist (cabergoline; Velactis, Ceva Santé Animale, Libourne, France; labeled for use only with abrupt dry-off, e.g., no preceding reduction in feeding level or milking frequency before last milking). After dry-off, all cows were fed the same dry cow diet and data collection continued for a week. Blood samples were collected from the coccygeal vein on d -9, -6, -5, -2, 1, 2, 5, and 7 relative to dry-off. Additionally, blood was sampled at 0, 3, and 6 h relative to injection of either cabergoline or saline, equivalent to d 0.125, 0.250, and 0.375 relative to last milking (dry-off). The reduced feeding level before dry-off caused reduced glucose and insulin concentrations as well as increased free fatty acid concentrations, particularly when reduced feeding level was combined with milking the cows 2× daily. The intramuscular injection of cabergoline caused the expected reduction in circulating prolactin concentrations. In addition, dopamine-agonist cabergoline induced an atypical simultaneous pattern of plasma metabolites (i.e., increased glucose and free fatty acid concentrations), hormones (i.e., reduced insulin and increased cortisol concentrations), and minerals (i.e., reduced calcium concentration), indicating that normal metabolic and mineral homeostatic regulations were hindered after the injection of ergot alkaloid cabergoline. In conclusion, reducing milking frequency seems the best management strategy to reduce milk production at dry-off among those tested in this study.

Effects of the density of extruded pellets on starch digestion kinetics, rumen fermentation, fiber digestibility and enteric methane production in dairy cows.

Dynamics of starch digestion in dairy cows fed extruded pellets differing in physical functional properties were investigated by measuring starch digestibility, post-prandial rumen fermentation patterns, and post-prandial duodenal starch appearance. Additionally, starch digestion effects on neutral detergent fiber (NDF) digestibility and methane (CH4 ) emission were studied. Pure barley was extruded to produce three treatments having pellets of either low-density (LD), medium-density (MD) or high-density (HD). The experiment was conducted in a 3 × 3 Latin square design using three lactating Danish Holstein cows fitted with ruminal, duodenal and ileal cannulas. After the allocation of experimental concentrate directly into the rumen through the rumen cannula, cows were fed a basal diet low in starch. Eight samples were collected on equal time intervals (9 h) from duodenal digesta, ileal digesta and feces (grab sample) to determine digestibility. For post-prandial rumen fermentation patterns, four sample sets of rumen dorsal, medial and ventral fluid were taken from each cow, whereas for post-prandial duodenal starch appearance, 14 samples of duodenal chyme were obtained from each cow relative to morning feeding of experimental concentrate at 07:00 h. Ruminal, small intestinal, hindgut and total tract digestibility of starch did not differ among treatments. Similarly, NDF digestibility and CH4 emission also remained unaffected by treatments. However, compared with the LD and MD treatments, the HD treatment showed higher acetate: propionate ratio at all positions in the rumen and a higher post-prandial duodenal starch appearance. This indicates lower ruminal starch degradation (RSD) and higher starch flow into the small intestine for HD treatment. In conclusion, the current study indicates that pellets' physical properties can manipulate RSD, where pellets with high density and fluid stability can partly shift starch digestion from the rumen to the small intestine. Indeed, further investigations are needed.

Is Cardiac Rehabilitation Equally Effective in Improving Dietary Intake in All Patients with Ischemic Heart Disease?

OBJECTIVE: To investigate the association between sociodemographic characteristics and changes in dietary intake in patients with ischemic heart disease who were following a cardiac rehabilitation program. METHODS: Longitudinal study among patients with first-time admission to outpatient cardiac rehabilitation after ST-elevation myocardial infarction, non-ST-elevation myocardial infarction, unstable angina pectoris or stable angina pectoris. We used a validated self-administered food frequency questionnaire to assess the dietary intake before rehabilitation, post rehabilitation and 6 months after baseline. The intake was summarized into a fat and fish-fruit-vegetable score for both sexes. To assess the within-patient differences in dietary scores, a paired t-test was applied. Regression analyses were performed to assess the association between sociodemographic characteristics and changes in diet. RESULTS: 186 patients completed two measurements, 157 patients completed all three measurements. Fat and fish-fruit-vegetable scores increased statistically significantly and improvements remained statistically significant at follow-up. Fat scores increased less in employed men than in retired men (-11 (95% CI -17; -5)). Fish-fruit-vegetable scores increased less in men with a bachelor degree than in men with a vocational education (-8 (95% CI -13; -3)) as well as in retired women compared with employed women (-18 (95% CI -32; -4)). CONCLUSIONS: Patients statistically significantly improved their dietary intake and improvements remained at follow-up. Dietary improvements were sensitive to marital status, living arrangements and employment status. Unemployed and retired women did not seem to improve their diet as much as employed women. Additional dietary intervention may become relevant in some patient groups defined by sociodemographic characteristics. (250).

Association between magnesium in drinking water and atrial fibrillation incidence: a nationwide population-based cohort study, 2002-2015.

BACKGROUND: Atrial fibrillation (AF) is a common heart rhythm disorder and a risk factor of adverse cardiovascular diseases. Established causes do not fully explain the risk of AF and unexplained risk factors might be related to the environment, e.g. magnesium in drinking water. Low magnesium levels in drinking water might be associated with higher risk of cardiovascular diseases including AF. With detailed individual data from nationwide registries and long-term magnesium exposure time series, we had a unique opportunity to investigate the association between magnesium in drinking water and AF. OBJECTIVE: We evaluated the association between magnesium concentration in drinking water and AF risk. METHODS: A nationwide register-based cohort study (2002-2015) was used including individuals aged ≥30 years. Addresses were linked with water supply areas (n = 2418) to obtain time-varying drinking water magnesium exposure at each address. Five exposure groups were defined based on a 5-year rolling time-weighted average magnesium concentration. AF incidence rate ratios (IRRs) between exposure groups were calculated using a Poisson regression of incidence rates, adjusted for sex, age, and socioeconomic position. Robustness of results was investigated with different exposure definitions. RESULTS: The study included 4,264,809 individuals (44,731,694 person-years) whereof 222,998 experienced an incident AF. Magnesium exposure ranged from 0.5 to 62.0 mg/L (mean = 13.9 mg/L). Estimated IRR (95% CI) compared to the referent exposure group (< 5 mg/L) was 0.98 (0.97-1.00) for the second lowest exposure group (5-10 mg/L), and 1.07 (1.05-1.08) for the two highest exposure groups (15-62 mg/L). Strongest positive associations were observed among those aged ≥80 years and with lowest education group. An inverse association was found among individuals with highest education group. CONCLUSION: There might be a small beneficial effect on AF of an increase in magnesium level in drinking water up to 10 mg/L, though an overall positive association was observed. The unexpected positive association and different associations observed for subgroups suggest a potential influence of unaccounted factors, particularly in vulnerable populations. Future research on magnesium in drinking water and cardiovascular diseases needs to focus on contextual risk factors, especially those potentially correlating with magnesium in drinking water.

Joint mapping of cardiovascular diseases: comparing the geographic patterns in incident acute myocardial infarction, stroke and atrial fibrillation, a Danish register-based cohort study 2014-15.

BACKGROUND: Disease mapping aims at identifying geographic patterns in disease. This may provide a better understanding of disease aetiology and risk factors as well as enable targeted prevention and allocation of resources. Joint mapping of multiple diseases may lead to improved insights since e.g. similarities and differences between geographic patterns may reflect shared and disease-specific determinants of disease. The objective of this study was to compare the geographic patterns in incident acute myocardial infarction (AMI), stroke and atrial fibrillation (AF) using the unique, population-based Danish register data. METHODS: Incident AMI, stroke and AF was modelled by a multivariate Poisson model including a disease-specific random effect of municipality modelled by a multivariate conditionally autoregressive (MCAR) structure. Analyses were adjusted for age, sex and income. RESULTS: The study included 3.5 million adults contributing 6.8 million person-years. In total, 18,349 incident cases of AMI, 28,006 incident cases of stroke, and 39,040 incident cases of AF occurred. Estimated municipality-specific standardized incidence rates ranged from 0.76 to 1.35 for AMI, from 0.79 to 1.38 for stroke, and from 0.85 to 1.24 for AF. In all diseases, geographic variation with clusters of high or low risk of disease after adjustment was seen. The geographic patterns displayed overall similarities between the diseases, with stroke and AF having the strongest resemblances. The most notable difference was observed in Copenhagen (high risk of stroke and AF, low risk of AMI). AF showed the least geographic variation. CONCLUSION: Using multiple-disease mapping, this study adds to the results of previous studies by enabling joint evaluation and comparison of the geographic patterns in AMI, stroke and AF. The simultaneous mapping of diseases displayed similarities and differences in occurrence that are non-assessable in traditional single-disease mapping studies. In addition to reflecting the fact that AF is a strong risk factor for stroke, the results suggested that AMI, stroke and AF share some, but not all environmental risk factors after accounting for age, sex and income (indicator of lifestyle and health behaviour).

Geographical variation and clustering are found in atrial fibrillation beyond socioeconomic differences: a Danish cohort study, 1987-2015.

BACKGROUND: The prevalence and incidence rate of atrial fibrillation (AF) increase worldwide and AF is a risk factor for more adverse cardiovascular diseases including stroke. Approximately 44% of AF cases cannot be explained by common individual risk factors and risk might therefore also be related to the environment. By studying geographical variation and clustering in risk of incident AF adjusted for socioeconomic position at an individual level, potential neighbourhood risk factors could be revealed. METHODS: Initially, yearly AF incidence rates 1987-2015 were estimated overall and stratified by income in a register-based cohort study. To examine geographical variation and clustering in AF, we used both spatial scan statistics and a hierarchical Bayesian Poisson regression analysis of AF incidence rates with random effect of municipalities (n = 98) in Denmark in 2011-2015. RESULTS: The 1987-2015 cohort included 5,453,639 individuals whereof 369,800 were diagnosed with an incident AF. AF incidence rate increased from 174 to 576 per 100,000 person-years from 1987 to 2015. Inequality in AF incidence rate ratio between highest and lowest income groups increased from 23% in 1987 to 38% in 2015. We found clustering and geographical variation in AF incidence rates, with incidence rates at municipality level being up to 34% higher than the country mean after adjusting for socioeconomic position. CONCLUSIONS: Geographical variations and clustering in AF incidence rates exist. Compared to previous studies from Alberta, Canada and the United States, we show that geographical variations exist in a country with free access to healthcare and even when accounting for socioeconomic differences at an individual level. An increasing social inequality in AF was seen from 1987 to 2015. Therefore, when planning prevention strategies, attention to individuals with low income should be given. Further studies focusing on identification of neighbourhood risk factors for AF are needed.

Effects of feeding level, milking frequency, and single injection of cabergoline on feed intake, milk yield, milk leakage, and clinical udder characteristics during dry-off in dairy cows.

Abrupt and gradual dry-off strategies by reducing feeding level (normal vs. reduced energy density), reducing milking frequency (twice vs. once daily), and administration of a dopamine agonist after last milking (i.m. saline vs. cabergoline injection) were investigated (2 × 2 × 2 factorial arrangement) for their effects on feed intake, milk yield, energy balance, milk leakage, and clinical udder characteristics in 119 Holstein cows. In the last week before dry-off, cows were assigned to 1 of 4 combinations of feeding level and milking frequency. Within 3 h after last milking, cows were injected with either saline or a dopamine agonist (cabergoline; Velactis, Ceva Santé Animale; labeled for use only with abrupt dry-off, i.e., no preceding reduction in feeding level or milking frequency before last milking). After dry-off, all cows were fed the same diet for dry cows, and data collection continued for a week. Dry matter intake (DMI) was recorded in automated feed bins and milk yield in an automatic milking system where additional concentrate was fed. Clinical udder characteristics and milk leakage were scored 10 times during the week before and the week after dry-off. Before dry-off, total DMI decreased with reduced feeding level compared with normal feeding level, but did not differ between milking frequencies. The combined effect of reduced DMI and diet energy concentration resulted in a 47% lower net energy intake with reduced feeding level compared with normal feeding level during the week before dry-off. Milk yield was approximately 30% lower during the week before dry-off when either feeding level or milking frequency was reduced compared with no change in feeding level or milking frequency, whereas milk yield was 45% lower when both feeding level and milking frequency were reduced. The net energy balance during the week before dry-off was negative with reduced feeding level and more negative when combined with twice-daily milking. After dry-off, udder engorgement was reduced in the 3 gradual dry-off treatments compared with abrupt dry-off. Cabergoline injection after last milking resulted in least udder engorgement and signs of milk leakage for 48 h, but also resulted in abrupt reduction of DMI lasting approximately 24 h irrespective of treatment before dry-off. In conclusion, gradual cessation of lactation by reducing milking frequency to once daily without reducing the feeding level decreased milk yield before dry-off in high-yielding dairy cows and reduced udder engorgement after dry-off without inducing negative energy balance during the period of dry-off. In contrast, reduced feeding level induced negative energy balance, which may compromise welfare due to metabolic stress and hunger. No clear differences in risk of milk leakage after dry-off were observed between abrupt and gradual dry-off management strategies. Use of cabergoline led to fewer signs of milk leakage and reduced udder engorgement during the first days after dry-off, which may positively affect welfare at dry-off. However, the mechanism behind and the welfare consequences of the concomitant abrupt decrease in DMI lasting approximately 24 h needs further investigation to complete our understanding of dopamine agonist use for dry-off.

Anxiety and depression symptoms in Danish patients with an implantable cardioverter-defibrillator: prevalence and association with indication and sex up to 2 years of follow-up (data from the national DEFIB-WOMEN study).

AIMS: To investigate (i) the prevalence of anxiety and depression and (ii) the association between indication for implantable cardioverter-defibrillator (ICD) implantation and sex in relation to anxiety and depression up to 24 months' follow-up. METHODS AND RESULTS: Patients with a first-time ICD, participating in the national, multi-centre, prospective DEFIB-WOMEN study (n = 1496; 18% women) completed the Hospital Anxiety and Depression Scale at baseline, 3, 6, 12, and 24 months. Data were analysed using linear mixed modelling for longitudinal data. Patients with a secondary prophylactic indication (SPI) had higher mean anxiety scores than patients with a primary prophylactic indication (PPI) at baseline, 3, and 12 months and higher mean depression scores at all-time points, except at 24 months. Women had higher mean anxiety scores as compared to men at all-time points; however, only higher mean depression scores at baseline. Overall, women with SPI had higher anxiety and depression symptom scores than men with SPI. Symptoms decreased over time in both women and men. From baseline to follow-up, the prevalence of anxiety (score ≥8) was highest in patients with SPI (13.3-20.2%) as compared to patients with PPI (range 10.0-14.7%). The prevalence of depression was stable over the follow-up period in both groups (range 8.5-11.1%). CONCLUSION: Patients with a SPI reported higher anxiety and depression scores as compared to patients with PPI. Women reported higher anxiety scores than men, but only higher depression scores at baseline. Women with SPI reported the highest anxiety and depression scores overall.

Lipid-lowering therapy and low-density lipoprotein cholesterol goal attainment after acute coronary syndrome: a Danish population-based cohort study.

BACKGROUND: Patients with acute coronary syndrome (ACS) are at high risk of recurrent cardiovascular (CV) event. The European guidelines recommend low-density lipoprotein cholesterol (LDL-C) levels < 1.8 mmol/L and early initiation of intensive lipid-lowering therapy (LLT) to reduce CV risk. In order to reduce the risk of further cardiac events, the study aimed to evaluate LDL-C goal attainment and LLT intensity in an incident ACS population. METHODS: A cohort study of patients with residency at Funen in Denmark at a first-ever ACS event registered within the period 2010-2015. Information on LLT use and LDL-C levels was extracted from national population registers and a Laboratory database at Odense University Hospital. Treatments and lipid patterns were evaluated during index hospitalization, at 6-month and 12-month follow-up. RESULTS: Among 3040 patients with an LDL-C measurement during index hospitalization, 40.7 and 39.0% attained the recommended LDL-C target value (< 1.8 mmol/L) within 6- and 12-month follow-up, respectively. During 6- and 12-month follow-up, a total of 89.2% (20.2%) and 88.4% (29.7%) used LLT (intensive LLT). Of the intensive LLT users, 43.4 and 47.7% reached the LDL-C target value at 6- and 12-month follow-up. The frequency of lipid monitoring was low: 69.5, 77.7 and 53.6% in patients with a first-ever ACS during index hospitalization, 6- and 12-month follow-up, respectively. CONCLUSION: Using national health registers and laboratory data, a considerably gap was observed between treatment guidelines and clinical practice in the management of dyslipidemia leaving very high-risk patients without adequate lipid management strategy. Therefore, improved lipid management strategies aimed at reaching treatment targets are warranted.

Technical note: Evaluation of the ororuminal FLORA sampling device for rumen fluid sampling in intact cattle.

The objectives of this study were to evaluate the actual intraruminal sampling site of the ororuminal FLORA sampling device (Profs Products, Wittibreut, Germany), and to compare pH and volatile fatty acid (VFA) data obtained using FLORA and those obtained using a suction strainer introduced to the ventral ruminal sac via a rumen cannula. Five lactating multiparous Danish Holstein cows fitted with ruminal cannulas were used. All cows were fed the same diet once daily, and the diet was allowed ad libitum. Samples of rumen fluid and recordings were obtained 6 h after feeding at 3 occasions. Rumen fluid samples were taken using 2 devices: (1) the ororuminal FLORA sampling device and (2) a suction strainer inserted through the rumen cannula to the ventral ruminal sac and a 60-mL syringe for suction. Both sampling devices were inserted concomitantly, and samples of rumen fluid were obtained simultaneously with both devices. After sampling rumen fluid, the actual intraruminal placement of the FLORA sampling cup was manually assessed as being in either the cranial sac, the dorsal sac, the medial layer, or the ventral sac. Only VFA proportions, and not pH and VFA concentrations, were similar between rumen fluid samples obtained using FLORA and those obtained directly through the rumen cannula. The observed intraruminal sampling site of the FLORA sampling cup indicates that firm rumen digesta hampers the ability of the FLORA device to reach the ventral sac and impedes filling of the sampling cup and, consequently, increases the risk of saliva contamination of samples.

Effects of toasting and decortication of oat on nutrient digestibility in the rumen and small intestine and on amino acid supply in dairy cows.

This study examined the potential for decorticating and toasting of oat (Avena sativa) to supply crude protein (CP) and amino acids (AA) in dairy cows. Four lactating Danish Holstein Friesian cows fitted with ruminal, duodenal, and ileal cannulas were assigned to a 4 × 4 Latin square design. Cows were fed experimental diets ad libitum based on grass-clover silage and toasted fava beans, with oat included in different forms arranged in a 2 × 2 factorial: whole oat, decorticated oat, toasted oat, and decorticated toasted oat. In situ rumen degradability of processed oat was also evaluated. Decortication increased starch intake by 0.38 kg/d and reduced NDF intake by 0.91 kg/d. Toasting reduced ruminal AA digestibility and increased duodenal flow of CP by 0.41 kg/d. In situ degradation rate and effective degradability of CP in the rumen were reduced by 0.46 h-1 and 310 g/kg CP due to toasting. Both decortication and toasting increased microbial synthesis of CP by 0.20 and 0.41 kg/d, respectively. Decortication and toasting did not affect small intestinal AA digestibility, but did increase the total digested amount of AA by 154 and 250 g/d, respectively. Milk production was not affected by treatments. Methane production (L/d) decreased with decortication and toasting. In conclusion, unless an interaction exists between decortication and toasting, the results indicate additive effects of toasting and decorticating oat for increasing the supply of digestible AA to the small intestine of dairy cows.

The influence of hydrolysis and derivatization on the determination of amino acid content and isotopic ratios in dual-labeled (13 C, 15 N) white clover.

RATIONALE: The cycling of peptide- and protein-bound amino acids (AAs) is important for studying the rate-limiting steps in soil nitrogen (N) turnover. A strong tool is stable C and N isotopes used in combination with compound-specific isotope analysis (CSIA), where a prerequisite for analysis is appropriate methods for peptide and protein hydrolysis and appropriate methods for derivatization of AAs for analysis by gas chromatography (GC). METHODS: We examined the efficiency of a standard acidic hydrolysis (6 M HCl, 20 h at 110°C) and a fast acidic hydrolysis (6 M HCl, 70 min at 150°C) on the recovery of AAs from a protein standard (bovine serum albumin). The best methods were used on dual-labeled (13 C and 15 N) clover shoot and root juice, divided into four molecular weight (Mw) size fractions. We used NAIP (N-acetyl isopropyl esterification) derivatization for GC/combustion-isotope ratio mass spectrometry (C-IRMS) analysis of AA standards. RESULTS: The NAIP derivatization gave very low limits of detection (LODs) (< 2 pmol) and limits of quantification (LOQs) ranging from 0.55 to 4.89 pmol. Comparing the concentrations of individual AAs in hydrolyzed versus unhydrolyzed clover juice samples of the low Mw size fraction (<1 kDa) showed a significant decline in concentration (p <0.03) for seven AAs after hydrolysis. Despite the decline in AA concentration, we found a linear connection between the obtained atomic fraction (13 C/total carbon and 15 N/total nitrogen) for individual AAs of hydrolyzed versus unhydrolyzed samples. CONCLUSIONS: The methodology distinguished differences in atomic fractions across AAs, in individual AAs in Mw size fractions, and between shoot and root samples of experimentally labeled white clover. Specifically, the method separated L-glutamate (Glu) and glutamine (Gln). Thus, for a broader use in plant and soil ecology, we present an optimized methodology for GC/C-IRMS analysis of AAs from organic nitrogen samples enriched with 13 C and 15 N - AA stable isotope probing (SIP).

Educational level and living arrangements are associated with dietary intake of red meat and fruit/vegetables: A Danish cross-sectional study.

Aims: To evaluate the association between education and living arrangements and the intake of fish, red meat and fruit and vegetables.Methods: The study design was cross-sectional and conducted in Denmark in 2013. Participants filled in questionnaires about their educational level and living arrangements (living alone or with others) and dietary intake including fish, red meat, fruit and vegetables. Regression analyses were performed to assess the associations within 85,456 randomly sampled healthy men and women who were at least 25 years old.Results: Length of education was statistically significant and positively associated with the intake of fruit and vegetables and negatively associated with the intake of red meat for both men and women. Men with a high level of education had a 187g/week (95% confidence interval: 199-175g/week) lower intake of red meat and a 109g/day (95% confidence interval: 102-117g/day) higher intake of fruit and vegetables than men with a low level of education. Women with a high level of education had a 175g/week (95% confidence interval: 186-164g/week) lower intake of red meat and a 106g/day (95% confidence interval: 97-114g/day) higher intake of fruit and vegetables than women with a low level of education. Living with others was statistically significant and positively associated with the intake of red meat, and fruit and vegetables. There were no clear associations between education, living arrangements and intake of fish.Conclusions: Men and women with a high educational level ate more fruit and vegetables but less red meat than men and women with a low educational level. Men and women living with others ate more red meat, fruit and vegetables than men and women living alone.

Medication adherence, biological and lifestyle risk factors in patients with myocardial infarction: a ten-year follow-up on socially differentiated cardiac rehabilitation.

Objective: There is strong evidence that medication adherence and lifestyle changes are essential in patients undergoing secondary cardiovascular disease prevention. Cardiac rehabilitation (CR) increases medication adherence and improves lifestyle changes. Patients with cardiac diseases and a low educational level and patients with little social support are less responsive to improve medication adherence and to adapt lifestyle changes. The aim of the present study was to investigate the long-term effects of a socially differentiated CR intervention on medication adherence as well as changes in biological and lifestyle risk factors at two- five- and ten-year follow-up. Design: A prospective cohort study. Setting: The cardiac ward at Aarhus University Hospital, Denmark. Intervention: A socially differentiated CR intervention in addition to the standard CR program. Subjects: Patients admitted with first-episode myocardial infarction between 2000 and 2004, N = 379. Patients were defined as socially vulnerable or non-socially vulnerable according to their educational level and extent of social network. Main outcome measures: Primary outcome was medication adherence to antithrombotics, beta-blockers, statins and angiotensin-converting enzyme inhibitors. Secondary outcomes were biological and lifestyle risk factors defined as; total cholesterol, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, glycated hemoglobin, blood pressure and smoking status. Results: No significant long-term effect of the intervention was found. Conclusions: The results indicate a non-significant effect of the intervention. However, it was found that equality in health was improved in the study population except concerning smoking. General practitioners manage to support the long-term secondary cardiovascular disease prevention in all patients regardless of social status. Key points The socially differentiated intervention did not significantly improve medication adherence or biological and lifestyle risk factors. Despite the non-significant effect of the intervention, equality in health was improved except concerning smoking. General practitioners managed to support the long-term secondary cardiovascular disease prevention in all patients regardless of social status.