Hepatic Global Transcriptomic Profiles of Holstein Cows According to Parity Reveal Age-Related Changes in Early Lactation.

Cows can live for over 20 years, but their productive lifespan averages only around 3 years after first calving. Liver dysfunction can reduce lifespan by increasing the risk of metabolic and infectious disease. This study investigated the changes in hepatic global transcriptomic profiles in early lactation Holstein cows in different lactations. Cows from five herds were grouped as primiparous (lactation number 1, PP, 534.7 ± 6.9 kg, n = 41), or multiparous with lactation numbers 2-3 (MP2-3, 634.5 ± 7.5 kg, n = 87) or 4-7 (MP4-7, 686.6 ± 11.4 kg, n = 40). Liver biopsies were collected at around 14 days after calving for RNA sequencing. Blood metabolites and milk yields were measured, and energy balance was calculated. There were extensive differences in hepatic gene expression between MP and PP cows, with 568 differentially expressed genes (DEGs) between MP2-3 and PP cows, and 719 DEGs between MP4-7 and PP cows, with downregulated DEGs predominating in MP cows. The differences between the two age groups of MP cows were moderate (82 DEGs). The gene expression differences suggested that MP cows had reduced immune functions compared with the PP cows. MP cows had increased gluconeogenesis but also evidence of impaired liver functionality. The MP cows had dysregulated protein synthesis and glycerophospholipid metabolism, and impaired genome and RNA stability and nutrient transport (22 differentially expressed solute carrier transporters). The genes associated with cell cycle arrest, apoptosis, and the production of antimicrobial peptides were upregulated. More surprisingly, evidence of hepatic inflammation leading to fibrosis was present in the primiparous cows as they started their first lactation. This study has therefore shown that the ageing process in the livers of dairy cows is accelerated by successive lactations and increasing milk yields. This was associated with evidence of metabolic and immune disorders together with hepatic dysfunction. These problems are likely to increase involuntary culling, thus reducing the average longevity in dairy herds.

Performance of Dairy Cows Offered Grass Silage Produced within Either a Three- or Four-Harvest System When Supplemented with Concentrates on a Feed-to-Yield Basis.

More frequent harvesting of grass swards provides an opportunity to improve the nutritive value of grass silage. This study investigated the effect of offering silages produced within either a three- (3H) or four-harvest (4H) system on dairy cow performance when concentrate supplements were offered according to the individual cow's milk yield (feed-to-yield). Cows (n = 80) were allocated to either 3H or 4H at calving and remained on experiment for 25 weeks. Within both treatments, cows were offered silage from each harvest consecutively in proportion to the dry matter (DM) yield for each harvest. Silage was offered as a mixed ration with concentrate added at a rate of 8 kg/cow/day. Additional concentrates were offered on a feed-to-yield basis. Herbage yields were reduced in the 4H system, but 4H silage contained higher metabolisable energy and crude protein content compared to 3H. Cows offered the 4H silage had greater silage DM intake, milk yield and milk protein content, while milk fat content was greater in cows offered 3H silages. In conclusion, increasing harvesting frequency from three to four harvests per year can improve silage feed value, silage intakes and milk yields when concentrates are offered on a feed-to-yield basis.

Milk Composition and Production Efficiency within Feed-To-Yield Systems on Commercial Dairy Farms in Northern Ireland.

This study examined the relationships between milk yield and diet composition, nutrient intakes, milk composition, and feed use efficiency when concentrates were offered using a feed-to-yield (FTY) approach. The study was conducted on 26 dairy farms in Northern Ireland. Cows (n = 3471) were fully housed and were offered concentrates on an FTY basis. Individual cow genetic information was obtained for 18 herds. Concentrate intakes of individual cows were either obtained from the farms or calculated, while milk yield and milk composition data were obtained from test-day milk recording. Mean test-day milk yields during months 2 to 5 post-calving were calculated for each cow, and cows within each lactation were placed into one of six equal-sized milk yield (kg/cow/day) groups. Diet effects and performance responses to milk yield groups were tested for linear and quadratic effects. Total dry matter intakes increased with increasing milk yield. Milk fat and milk protein concentration declined as milk yield increased, which could be attributed in part to genetics and diet. As milk yield increased, nitrogen and energy use efficiency was improved. However, concentrates offered per kg of energy-corrected milk also increased at higher milk yields, indicating an increased reliance on concentrates for these cows.

Impact of adopting non-antibiotic dry-cow therapy on the performance and udder health of dairy cows.

BACKGROUND: On dairy farms, the prophylactic use of antibiotics at drying-off is being increasingly challenged. The objective of this study was to examine the effect of antibiotic dry-cow therapy (DCT) or non-antibiotic DCT on dairy cow performance and udder health. METHODS: Holstein cows (n = 285) with low risk of intramammary infection (<200,000 cells/ml) were assigned to one of two treatments, either antibiotic DCT (A + TS; antibiotic treatment in combination with internal and external teat sealants) or non-antibiotic DCT (TS; internal and external teat sealant only). RESULTS: There was no statistically significant (p > 0.05) difference between treatments for mean cow milk yield, composition or energy corrected milk yield. Mean somatic cell count was 0.16 loge higher in the TS treatment (95% confidence interval [CI]: -0.00 loge to -0.33 loge ) compared to A + TS treatment (p = 0.047). A 50% increase in the number of mastitis cases was observed in the A + TS treatment compared to TS treatment (odds ratio = 1.5, 95% CI: 0.80%-3.01%), although this was not significant. There was no statistical evidence (p > 0.05) that treatment had any effect on colostrum quality and composition. CONCLUSION: Results indicate that non-antibiotic DCT can be adopted in 'low-risk' cows who were offered grass silage-based diets in cubicle accommodation, with low risk of adverse effects on performance or udder health.

Pasture access and eye temperature in dairy cows.

Pasture access can benefit dairy cows' behavior, health, and welfare, but herds are increasingly housed indoors full-time. Recent infrared thermal-imaging (thermography) studies suggest that higher eye temperatures may be a physiological indicator of chronic stress. We, therefore, hypothesized that, compared to cows with pasture access, cows housed indoors full-time would have higher eye temperatures. In a two-phase crossover experiment, 29 Holstein-Friesian dairy cows experienced 18 days of overnight pasture access and 18 days of full-time indoor housing. We measured each animal's eye temperature 16 times (eight/phase). During Phase One, cows with pasture access had higher eye temperatures than cows housed indoors full-time (contrary to our hypothesis). However, during Phase Two, cows with pasture access had lower eye temperatures than cows housed indoors full-time. It is, therefore, unclear whether eye temperature reflected disparities in dairy cow welfare between different housing treatments.

Proportion of Concentrate in the Diet of Early Lactation Dairy Cows Has Contrasting Effects on Circulating Leukocyte Global Transcriptomic Profiles, Health and Fertility According to Parity.

The functionality of circulating leukocytes in dairy cows is suppressed after calving, with negative energy balance as a risk factor. Leukocyte transcriptomic profiles were compared separately in 44 multiparous (MP) and 18 primiparous (PP) Holstein-Friesian cows receiving diets differing in concentrate proportion to test whether immune dysfunction could be mitigated by appropriate nutrition. After calving, cows were offered either (1) low concentrate (LC); (2) medium concentrate (MC) or (3) high concentrate (HC) diets with proportions of concentrate to grass silage of 30%:70%, 50%:50% and 70%:30%, respectively. Cow phenotype data collected included circulating metabolites, milk yield and health and fertility records. RNA sequencing of circulating leukocytes at 14 days in milk was performed. The HC diet improved energy balance in both age groups. There were more differentially expressed genes in PP than MP cows (460 vs. 173, HC vs. LC comparison) with few overlaps. The MP cows on the LC diet showed upregulation of the complement and coagulation cascade and innate immune defence mechanisms against pathogens and had a trend of more cases of mastitis and poorer fertility. In contrast, the PP cows on the HC diet showed greater immune responses based on both gene expression and phenotypic data and longer interval of calving to conception. The leukocytes of MP and PP cows therefore responded differentially to the diets between age, nutrient supply and immunity affecting their health and subsequent fertility.

Revisiting the Relationships between Fat-to-Protein Ratio in Milk and Energy Balance in Dairy Cows of Different Parities, and at Different Stages of Lactation.

A statistical re-assessment of aggregated individual cow data was conducted to examine trends in fat-to-protein ratio in milk (FPR), and relationships between FPR and energy balance (EB, MJ of ME/day) in Holstein-Friesian dairy cows of different parities, and at different stages of lactation. The data were collected from 27 long-term production trials conducted between 1996 and 2016 at the Agri-Food and Biosciences Institute (AFBI) in Hillsborough, Northern Ireland. In total, 1321 lactations (1 to 20 weeks in milk; WIM), derived from 840 individual cows fed mainly grass silage-based diets, were included in the analysis. The energy balance was calculated daily and then averaged weekly for statistical analyses. Data were further split in 4 wk. intervals, namely, 1-4, 5-8, 9-12, 13-16, and 17-20 WIM, and both partial correlations and linear regressions (mixed models) established between the mean FPR and EB during these periods. Three FPR score categories ('Low' FPR, <1.0; 'Normal' FPR, 1.0-1.5; 'High' FPR, >1.5) were adopted and the performance and EB indicators within each category were compared. As expected, multiparous cows experienced a greater negative EB compared to primiparous cows, due to their higher milk production relative to DMI. Relatively minor differences in milk fat and protein content resulted in large differences in FPR curves. Second lactation cows displayed the lowest weekly FPR, and this trend was aligned with smaller BW losses and lower concentrations of non-esterified fatty acids (NEFA) until at least 8 WIM. Partial correlations between FPR and EB were negative, and 'greatest' in early lactation (1-4 WIM; r = -0.38 on average), and gradually decreased as lactation progressed across all parities (17-20 WIM; r = -0.14 on average). With increasing parity, daily EB values tended to become more negative per unit of FPR. In primiparous cows, regression slopes between FPR and EB differed between 1-4 and 5-8 WIM (-54.6 vs. -47.5 MJ of ME/day), while differences in second lactation cows tended towards significance (-57.2 vs. -64.4 MJ of ME/day). Irrespective of the lactation number, after 9-12 WIM, there was a consistent trend for the slope of the linear relationships between FPR and EB to decrease as lactation progressed, with this likely reflecting the decreasing milk nutrient demands of the growing calf. The incidence of 'High' FPR scores was greatest during 1-4 WIM, and decreased as lactation progressed. 'High' FPR scores were associated with increased energy-corrected milk (ECM) yields across all parities and stages of lactation, and with smaller BW gains and increasing concentrations (log transformed) of blood metabolites (non-esterified fatty acid, NEFA; beta-hydroxybutyrate, BHB) until 8 WIM. Results from the present study highlight the strong relationships between FPR in milk, physiological changes, and EB profiles during early lactation. However, while FPR can provide an indication of EB at a herd level, the large cow-to-cow variation indicates that FPR cannot be used as a robust indicator of EB at an individual cow level.

Optimism and pasture access in dairy cows.

Allowing dairy cattle to access pasture can promote natural behaviour and improve their health. However, the psychological benefits are poorly understood. We compared a cognitive indicator of emotion in cattle either with or without pasture access. In a crossover experiment, 29 Holstein-Friesian dairy cows had 18 days of overnight pasture access and 18 days of full-time indoor housing. To assess emotional wellbeing, we tested cows on a spatial judgement bias task. Subjects learnt to approach a rewarded bucket location, but not approach another, unrewarded bucket location. We then presented cows with three "probe" buckets intermediate between the trained locations. Approaching the probes reflected an expectation of reward under ambiguity-an "optimistic" judgement bias, suggesting positive emotional states. We analysed the data using linear mixed-effects models. There were no treatment differences in latency to approach the probe buckets, but cows approached the known rewarded bucket slower when they had pasture access than when they were indoors full-time. Our results indicate that, compared to cattle housed indoors, cattle with pasture access display less anticipatory behaviour towards a known reward. This reduced reward anticipation suggests that pasture is a more rewarding environment, which may induce more positive emotional states than full-time housing.

Proxy Measures and Novel Strategies for Estimating Nitrogen Utilisation Efficiency in Dairy Cattle.

The efficiency with which dairy cows convert dietary nitrogen (N) to milk N is generally low (typically 25%). As a result, much of the N consumed is excreted in manure, from which N can be lost to the environment. Therefore there is increasing pressure to reduce N excretion and improve N use efficiency (NUE) on dairy farms. However, assessing N excretion and NUE on farms is difficult, thus the need to develop proximate measures that can provide accurate estimates of nitrogen utilisation. This review examines a number of these proximate measures. While a strong relationship exists between blood urea N and urinary N excretion, blood sampling is an invasive technique unsuitable for regular herd monitoring. Milk urea N (MUN) can be measured non-invasively, and while strong relationships exist between dietary crude protein and MUN, and MUN and urinary N excretion, the technique has limitations. Direct prediction of NUE using mid-infrared analysis of milk has real potential, while techniques such as near-infrared spectroscopy analysis of faeces and manure have received little attention. Similarly, techniques such as nitrogen isotope analysis, nuclear magnetic resonance spectroscopy of urine, and breath ammonia analysis may all offer potential in the future, but much research is still required.

Modern Holstein-origin dairy cows within grassland-based systems partition more feed nitrogen into milk and excrete less in manure.

The objective was to determine whether modern Holstein-origin dairy cows, when managed within grassland-based systems, partitioned more feed nitrogen (N) into milk and excreted less in manure, in comparison to an earlier population of Holstein-origin dairy cows. Data used were collated from total diet digestibility studies undertaken in Northern Ireland from 1990 to 2002 (old dataset, n = 538) and from 2005 to 2019 (new dataset, n = 476), respectively. An analysis of variance indicated that cows in the new dataset partitioned a significantly higher proportion of consumed N into milk and excreted a lower proportion in urine and total manure, compared to cows in the old dataset. A second analysis using the linear regression revealed that in comparison to the old dataset, the new dataset had a lower slope in the relationship between N intake and N excretion in urine or total manure, while a higher slope in the relationship between N intake and milk N output. A third analysis used the combined data from both datasets to examine if there was a relationship between experimental year and N utilization efficiency. Across the period from 1990 to 2019, urine N/N intake and manure N/N intake significantly decreased, while milk N/N intake increased. These results indicate that modern Holstein-origin dairy cows utilize consumed N more efficiently than earlier populations. Thus, N excretion is likely to be overestimated if models developed from the old data are used to predict N excretion for modern dairy herds. Therefore, the final part of analysis involved using the new dataset to develop prediction models for N excretion based on N intake and farm level data (milk yield, live weight and dietary N concentration). These updated models can be used to estimate N excretion from modern Holstein-origin dairy cows within grassland-based dairy systems.

Pasture Access Affects Behavioral Indicators of Wellbeing in Dairy Cows.

Dairy cows are increasingly housed indoors, either year-round or for long stretches over the winter and around parturition. This may create health and welfare issues. In cattle, lying and walking are highly motivated, and herds synchronize lying behavior when they have comfortable surfaces and little competition for space. Lying and walking activity can, therefore, indicate good welfare. Using a repeated measures crossover design, we gave 29 Holstein-Friesian dairy cows 18 days of overnight pasture access (PAS treatment) and 18 days of indoor housing (PEN treatment). Accelerometers recorded their lying and locomotory behavior. We measured behavioral synchrony with Fleiss' Kappa and analyzed the accelerometry data using linear mixed models. Compared to the PEN treatment, the PAS treatment had longer overnight lying durations (χ21 = 27.51, p < 0.001), fewer lying bouts (χ21 = 22.53, p < 0.001), longer lying bouts (χ21 = 25.53, p < 0.001), and fewer transitions up or down (χ21 = 16.83, p < 0.001). Herd lying behavior was also more synchronous at pasture (χ21 = 230.25, p < 0.001). In addition, nightly step counts were higher in the PAS treatment than the PEN treatment (χ21 = 2946.31, p < 0.001). These results suggest pasture access improves dairy cow welfare by increasing comfort, reducing competition and boredom, and facilitating motivated behavior.

Comparison of total-mixed-ration and feed-to-yield strategies on blood profiles and dairy cow health.

Seventy-two Holstein-Friesian dairy cows were offered the same amount of concentrates over the first 140 days of lactation, by either a 'total-mixed-ration' or a 'feed-to-yield' strategy. The effects on blood profiles and cow health were examined. Cows on total-mixed-ration were offered a mixed ration comprising grass silage and concentrates (50:50 dry matter basis). Cows on feed-to-yield were offered a basal mixed ration (grass silage plus 6 kg concentrates/cow/day) plus additional concentrates via an out-of-parlour feeding system, calculated according to each individual cow's milk yield during the previous week. Cows on total-mixed-ration had a higher mean haemoglobin, packed cell volume and lymphocyte percentage. Concentrate allocation strategy had no effect on serum haptoglobin concentrations, interferon-gamma production of pokeweed mitogen-stimulated whole blood culture, the incidence of clinical or subclinical mastitis, lameness, respiratory or digestive problems and no strong relationships were identified between production parameters with serum metabolites, inflammatory and immune measures. This study demonstrates small physiological differences in metabolic parameters, and no differences in inflammatory or immune parameters, when allocating concentrates by total-mixed-ration or feed-to-yield.