Soil plays a key role in the global carbon (C) cycle. Most current assessments of SOC stocks and the guidelines given by Intergovernmental Panel on Climate Change (IPCC) focus on the top 30 cm of soil. Our research shows that, when considering only total quantities, most of the SOC stocks are found in this top layer. However, not all forms of SOC are equally valuable as long-term stable stores of carbon: the majority of SOC is available for mineralisation and can potentially be re-emitted to the atmosphere. SOC associated with micro-aggregates and silt plus clay fractions is more stable and therefore represents a long-term carbon store. Our research shows that most of this stable carbon is located at depths below 30 cm (42% of subsoil SOC is located in microaggregates and silt and clay, compared to 16% in the topsoil), specifically in soils that are subject to clay illuviation. This has implications for land management decisions in temperate grassland regions, defining the trade-offs between primary productivity and C emissions in clay-illuviated soils, as a result of drainage. Therefore, climate smart land management should consider the balance between SOC stabilisation in topsoils for productivity versus sequestration in subsoils for climate mitigation.

Bifidobacteria play a vital role in human nutrition and health by shaping and maintaining the gut ecosystem. In order to exert a beneficial effect, a sufficient population of bifidobacteria must colonise the host. In this study, we developed a miniaturised high-throughput in vitro assay for assessing the colonising ability of bacterial strains in human cells. We also investigated a variety of components isolated from different milk sources for their ability to increase the adherence of Bifidobacterium longum subsp. infantis ATCC 15697, a common member of the gastrointestinal microbiota of breastfed infants, to HT-29 cells. Both conventional and miniaturised colonisation assays were employed to examine the effect of 13 different milk-derived powders on bacterial adherence, including positive controls which had previously resulted in increased bifidobacterial adherence (human milk oligosaccharides and a combination of 3′- and 6′-sialylactose) to intestinal cells. Immunoglobulin G enriched from bovine whey and goat milk oligosaccharides resulted in increased adhesion (3.3- and 8.3-fold, respectively) of B. infantis to the intestinal cells and the miniaturised and conventional assays were found to yield comparable and reproducible results. This study highlights the potential of certain milk components to favourably modulate adhesion of bifidobacteria to human intestinal cells.

RNAlater® is regarded as a potential preservation method for proteins, while its effect on bovine muscle proteins has rarely been evaluated. Bovine muscle protein samples (n = 12) collected from three tender (Warner–Bratzler shear force: 30.02–31.74 N) and three tough (Warner–Bratzler shear force: 54.12–66.25 N) Longissimus thoracis et lumborum (LTL) samples, preserved using two different sampling preservation methods (RNAlater® and dry ice), at two post mortem time points (day 0 and day 14), were characterized using one-dimensional electrophoresis. Fourteen bands with molecular weights ranging from 15 to 250 kDa were verified, both in the dry ice and RNAlater® storage groups, at each time point, using image analysis. A shift from high to low molecular weight fragments, between day 0 and day 14, indicated proteolysis of the muscle proteins during post mortem storage. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analyses and database searching resulted in the identification of 10 proteins in four bands. Protein profiles of muscle preserved in RNAlater® were similar to those of muscle frozen on dry ice storage, both at day 0 and day 14. The results demonstrate that RNAlater® could be a simple and efficient way to preserve bovine muscle proteins for bovine muscle proteomic studies

Infection of cattle with bovine paratuberculosis (i.e., Johne's disease) is caused by Mycobacterium avium ssp. paratuberculosis (MAP) and results in a chronic incurable gastroenteritis. This disease, which has economic ramifications for the cattle industry, is increasing in detected prevalence globally; subclinically infected animals can silently shed the bacterium into the environment for years, exposing contemporaries and hampering disease-control programs. The objective of the present study was to first quantify the genetic parameters for humoral response to MAP in dairy cattle. This was followed by a genome-based association analysis and subsequent downstream bioinformatic analyses from imputed whole genome sequence SNP data. After edits, ELISA test records were available on 136,767 cows; analyses were also undertaken on a subset of 33,818 of these animals from herds with at least 5 MAP ELISA-positive cows, with at least 1 of those positive cows being homebred. Variance components were estimated using univariate animal and sire linear mixed models. The heritability calculated from the animal model for humoral response to MAP using alternative phenotype definitions varied from 0.02 (standard error = 0.003) to 0.05 (standard error = 0.008). The genome-based associations were undertaken within a mixed model framework using weighted deregressed estimated breeding values as a dependent variable on 1,883 phenotyped animals that were ≥87.5% Holstein-Friesian. Putative susceptibility quantitative trait loci (QTL) were identified on Bos taurus autosome 1, 3, 5, 6, 8, 9, 10, 11, 13, 14, 18, 21, 23, 25, 26, 27, and 29; mapping the most significant SNP to genes within and overlapping these QTL revealed that the most significant associations were with the 10 functional candidate genes KALRN, ZBTB20, LPP, SLA2, FI3A1, LRCH3, DNAJC6, ZDHHC14, SNX1, and HAS2. Pathway analysis failed to reveal significantly enriched biological pathways, when both bovine-specific pathway data and human ortholog data were taken into account. The existence of genetic variation for MAP susceptibility in a large data set of dairy cows signifies the potential of breeding programs for reducing MAP susceptibility. Furthermore, the identification of susceptible QTL facilitates greater biological understanding of bovine paratuberculosis and potential therapeutic targets for future investigation. The novel molecular similarities identified between bovine paratuberculosis and human inflammatory bowel disease suggest potential for human therapeutic interventions to be translated to veterinary medicine and vice versa.

Phosphorus (P) loss from soils to water can be a major pressure on freshwater quality and dairy farming, with higher animal stocking rates, may lead to potentially greater nutrient source pressures. In many countries with intensive agriculture, regulation of P management aims to minimise these losses. This study examined the P transfer continuum, from source to impact, in a dairy-dominated, highly stocked, grassland catchment with free-draining soils over three years. The aim was to measure the effects of P source management and regulation on P transfer across the nutrient transfer continuum and subsequent water quality and agro-economic impacts. Reduced P source pressure was indicated by: (a) lower average farm-gate P balances (2.4 kg ha−1 yr−1), higher P use efficiencies (89%) and lower inorganic fertilizer P use (5.2 kg ha−1 yr−1) relative to previous studies; (b) almost no recorded P application during the winter closed period, when applications were prohibited, to avoid incidental transfers; and (c) decreased proportions of soils with excessive P concentrations (32–24%). Concurrently, production and profitability remained comparable with the top 10% of dairy farmers nationally with milk outputs of 14,585 l ha−1, and gross margins of € 3130 ha−1. Whilst there was some indication of a response in P delivery in surface water with declines in quick flow and interflow pathway P concentrations during the winter closed period for P application, delayed baseflows in the wetter third year resulted in elevated P concentrations for long durations and there were no clear trends of improving stream biological quality. This suggests a variable response to policy measures between P source pressure and delivery/impact where the strength of any observable trend is greater closer to the source end of the nutrient transfer continuum and a time lag occurs at the other end. Policy monitoring and assessment efforts will need to be cognisant of this.

White clover (Trifolium repens) is an important legume for grazed grassland that can increase the profitability and environmental sustainability of milk production. Previous experiments on mown grass-clover plots suggest that low postgrazing heights (PGH) can increase sward clover content and herbage production. However, this has not been tested in actual strip or rotational grazing systems with dairy cows. Furthermore, lowering PGH in grass-only swards (typically perennial ryegrass without white clover) has previously been associated with reduced milk yields per cow. The objective of this experiment was to investigate the effect of PGH by dairy cows on clover content, herbage production, and milk production from strip-grazed grass-white clover swards in Ireland. Three target PGH treatments of 4, 5, and 6 cm were in place for entire grazing seasons (February to November) for 3 consecutive years (2007 to 2009). Each treatment had a mean of 21 Holstein-Friesian dairy cows that strip-grazed a mean annual area of 10.2 ha. Postgrazing height was measured twice a day with a rising plate meter, and cows were moved to the next strip once the target PGH was reached. Annual fertilizer nitrogen input was 90 kg of N/ha for each treatment. The PGH treatment did not significantly affect annual milk yield (6,202 kg/cow), solids-corrected milk yield (6,148 kg/cow), fat, protein, or lactose yields (265, 222, and 289 kg/cow, respectively), cow liveweight (592 kg) or body condition score (3.01). The PGH treatment also had no significant effect on sward white clover content (196 g/kg). However, herbage production of both grass and clover were significantly higher with the 4-cm PGH treatment compared with the 6-cm treatment. Mean annual herbage yields were 11.1, 10.2, and 9.1 t of organic matter (OM)/ha for the 4-, 5-, and 6-cm PGH treatments, respectively. The lower herbage production in the 6-cm PGH treatment resulted in lower annual silage production, greater housing requirements, and a substantially higher net silage deficit (−1,917 kg of OM/cow) compared with the 5- or 4-cm treatments (−868 and −192 kg of OM/cow, respectively). Grazing to a PGH of 4 cm is therefore recommended for grass-white clover swards.

Inflammation of the post-partum uterus is a normal physiological event, crucial for tissue involution and repair. However, in the bovine, some cows fail to resolve this inflammation, resulting in endometritis, which compromises fertility. Earlier work has identified upregulated expression of the potent inflammatory cytokine IL-1β early post-partum, in cows which subsequently develop endometritis. As a result, targeting IL-1β expression holds potential as a novel treatment for this disease, yet the regulatory mechanisms contributing to IL-1β expression in the bovine endometrium remain unknown. To investigate this, endometrial tissue samples were obtained 7 and 21 days post-partum (DPP) from cows that were diagnosed with endometritis at 21 DPP and cows that experienced a physiological level of inflammation throughout involution. IL-1β was measured by qPCR, ELISA, and immunohistochemistry. Seven DPP, endometrial IL-1β protein levels were significantly higher in animals that proceeded to develop endometritis at 21 DPP. IL-1β production could be detected in luminal and glandular epithelium, in underlying stromal fibroblasts as well as infiltrating immune cells. To investigate the mechanisms regulating IL-1β expression, primary endometrial epithelial cells, stromal fibroblasts and PBMCs were stimulated with LPS and the inflammasome activator nigericin. Stromal fibroblast cells were particularly potent producers of IL-1β. Basolateral LPS stimulation of polarized epithelial cells induced IL1B mRNA and a previously undescribed IL-1β protein isoform, with preferential protein secretion into the apical compartment. Key inflammasome components [nod-like receptor protein 3 (NLRP3), nima-related kinase-7 (NEK7), apoptosis speck like protein containing a CARD (ASC), and gasdermin-D] were expressed by endometrial cells following stimulation. Endometrial cell stimulation in the presence of NLRP3 receptor (MCC950) and pan-caspase (Z-VAD-FMK) inhibitors blocked IL-1β production, demonstrating its dependence on the NLRP3 inflammasome and on caspase activity. Furthermore, caspase-4 specific siRNA prevented IL-1β production, confirming that inflammasome activation in endometrial cells is caspase-4 but not caspase-1 dependent, as shown in other species. Identifying the tissue- and species-specificity of inflammasome assembly and activation has critical relevance for our understanding of inflammation and suggests new therapeutic targets to enhance the resolution of inflammatory pathologies including endometritis in cattle.

Bovine viral diarrhoea virus (BVDV) and bovine herpesvirus 1 (BoHV-1) are contagious bovine viral agents. The objectives of this study were to use quarterly bulk milk and ‘spot’ testing of unvaccinated youngstock to establish the national prevalence of exposure to BVDV and/or BoHV-1 in Irish dairy herds. Seasonality of bulk milk ELISA results was also examined. From a geographically representative population of 305 dairy herds, 88% and 80% of herds yielded mean annual positive bulk milk readings for BVDV and BoHV-1, respectively. Of these, 61% were vaccinated against BVDV and 12% against BoHV-1. A total of 2171 serum samples from weanlings having a mean age of 291 days yielded 543 (25%) seropositive for BVDV, and 117 (5.4%) seropositive for BoHV-1. A significant seasonal trend in bulk milk antibody ELISA readings and herd status was recorded for BVDV, with more herds categorised as positive in the latter half of the year.

Grazed grass is the cheapest feed available for dairy cows in temperate regions; thus, to maximize profits, dairy farmers must optimize the use of this high-quality feed. Previous research has defined the benefits of including white clover (Trifolium repens L.) in grass swards for milk production, usually at reduced nitrogen usage and stocking rate. The aim of this study was to quantify the responses in milk production of dairy cows grazing tetraploid or diploid perennial ryegrass (Lolium perenne L.; PRG) sown with and without white clover but without reducing stocking rate or nitrogen usage. We compared 4 grazing treatments in this study: tetraploid PRG-only swards, diploid PRG-only swards, tetraploid with white clover swards, and diploid with white clover swards. Thirty cows were assigned to each treatment, and swards were rotationally grazed at a farm-level stocking rate of 2.75 cows/ha and a nitrogen fertilizer rate of 250 kg/ha annually. Sward white clover content was 23.6 and 22.6% for tetraploid with white clover swards and diploid with white clover swards, respectively. Milk production did not differ between the 2 ploidies during this 4-yr study, but cows grazing the PRG-white clover treatments had significantly greater milk yields (+596 kg/cow per year) and milk solid yields (+48 kg/cow per year) compared with cows grazing the PRG-only treatments. The PRG-white clover swards also produced 1,205 kg of DM/ha per year more herbage, which was available for conserving and buffer feeding in spring when these swards were less productive than PRG-only swards. Although white clover is generally combined with reduced nitrogen fertilizer use, this study provides evidence that including white clover in either tetraploid or diploid PRG swards, combined with high levels of nitrogen fertilizer, can effectively increase milk production per cow and per hectare.

The application of natural preservatives on fresh fish has potential to extend shelf-life. In the present study, 8 essential oils (EOs) (lemon, lemongrass, lime, garlic, onion, oregano, thyme and rosemary) and 3 organic acids (OAs) (ascorbic, citric and lactic) were evaluated. The antimicrobial activity of these compounds was tested in-vitro against four confirmed Listeria spp. isolated from retail skin-packed salmon and cod. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were established for each compound. Then, a sensory evaluation was performed by a panel of ‘expert assessors’ on cooked fish treated with all of the OAs and any 4 EOs with a MIC <0.8%. A series of descriptors were assigned to characterize the combination of each compound with cooked salmon or cod. The highest antimicrobial effect against all Listeria spp. was observed for lactic acid (0.31–2.5%), but treatment with this compound resulted in the development of organoleptically unacceptable changes in salmon or cod. The most acceptable OAs for salmon and cod were ascorbic acid (1.25%) and citric acid (0.63%) respectively, which were shown to enhance certain organoleptic characteristics. The most effective EO against all Listeria strains evaluated was oregano oil (0.2%) and it was considered suitable as a treatment for salmon. In contrast, none of the EOs tested was organoleptically acceptable in combination with cod because of their strong odours and flavours that masked the fresh attributes associated with this fish.