This study determined the effect of feeding synthetic zeolite A prepartum to reduce periparturient hypocalcaemia risk on the eating, lying, and activity behaviours of multiparous grazing dairy cows. Animals were randomly allocated to either a Zeolite treatment group (n = 21; individually fed 500 g/d zeolite for two weeks prepartum) or a Control group (n = 22). Behaviour data obtained from accelerometers were analysed to determine the effects of treatment, parity (Parity 2-3 vs. 4+), and their interactions during 3 periods: PRE (-21 to -3 d), PERI (-2-2 d) and POST (3-28 d) relative to the day of calving (day 0). Parity 2-3 Zeolite cows ate for 18 min/day less than Parity 2-3 Control cows PRE. Zeolite-fed cows also ate for 24 min/day less than Control cows during the PERI period. There were no treatment differences in lying time or activity PRE or PERI, but Zeolite-fed cows spent more time lying POST. Regardless of treatment, Parity 2-3 cows were generally more active and ate for longer than Parity 4 + cows. Our results suggest zeolite may subtly decrease appetite or alter eating behaviour during the supplementation period, particularly in younger cows, and may improve cow comfort postcalving.

In grazing systems, feeds other than pasture are often used to increase dry matter intake and milk production. The study aimed to evaluate the effect of individually feeding pasture-grazed dairy cows near peak lactation based on their individualised milk production compared with a flat-rate feeding regime. All cows in the flat-rate treatment were offered 4 kg wet weight (3.6 kg DM) of concentrate/d. In the individualised feeding treatment, cows were offered 6, 4, and 2 kg wet weight/d (5.4, 3.6 and 1.8 kg DM/d) in the high, moderate, and low production groups, respectively. Individualised feeding of concentrates based on cow milk production did not alter milk or milk component yield at a herd level, but increased milk yield and tended to increase yield of fat, protein, and lactose in cows in the medium milk yield tertile. Individualised feeding reduced milk protein yield in cows in the low milk yield tertile, compared with flat rate feeding. The results do not support individualised feeding in grazing dairy systems where the maximum level of concentrate offered/ is <= 6 kg DM/d and milk production/cow <= 30 kg milk/d or <= 2.5 kg fat and protein/d.

The Number 2 Dairy (No. 2 Dairy) unit at Ruakura, Hamilton, New Zealand was established as a dairy research farm in 1943 and became recognised globally as one of the leading institutes for farm systems science. The research undertaken was fundamental to the 'systemisation' of component research into replicable, efficient, competitive, and sustainable dairy systems. From McMeekan's seminal research in the 1950s on rotational grazing to the experiments of Carter, Campbell, Bryant, Penno and others, who brought greater resolution to the key principles influencing successful outcomes on-farm, the more than 60 years of experimental research led to 'good practice' rules that were to transcend geographical, climatic, and system-level interactions. Much of the research can only be found in Annual Reports or in summary form in conference proceedings and not in an electronic form. Our objective was to collate and discuss the research undertaken, thereby ensuring that the legacy of this great work and the wisdom amassed are not lost. To this end, we have compiled a compendium of the research undertaken at No. 2 Dairy, with the hope that it be used to inform the scientific rationale for system design and to stimulate future hypotheses for system improvement.

Grazing cows undergo a similar degree of metabolic stress and immune dysregulation to those reported in high-yielding housed cows consuming total mixed rations, but the ability to manage daily nutrient intake is much less. Feed quality varies from day-to-day and weather can greatly influence amount consumed on any given day. Transition cow management, therefore, tends to revolve around pragmatism as opposed to precision. Mid- and late-gestation management of body condition score is essential to a smooth transition period.

The efficacy of a long-acting synthetic derivative of kisspeptin (Kp) to initiate normal oestrous cycles was tested in 24 mixed-aged, Holstein-Friesian cows that were 18-25 days postpartum on the day of treatment (D0). Groups of eight cows received saline (Sal) vehicle by intramuscular injection at 8:00 and 16:00 h (Sal-Sal), Kp at 8:00 h and vehicle at 16:00 h (Kp-Sal) or Kp on both occasions (Kp-Kp). The Kp dose was 15 nmol per 60 kg body weight. The ovaries of the cows were examined daily by ultrasonography between D4 and D14. Blood samples were collected from a tail vessel at 0, 2, 4, 8, 10 and 12 h relative to the time of the first injection for luteinizing hormone (LH) and follicle-stimulating hormone assay. Additional samples were collected daily from D4 until D14 and D19, 22, 26 and 29 for progesterone assay. LH surge-like responses were observed in cows treated with Kp at 8:00 h. Ovulation was consistently induced by Kp within 48 h when a dominant ovarian follicle of at least 10 mm in diameter was observed (8/14) but in no cases (6/14) during a new wave of ovarian follicular development comprising follicles <10 mm in diameter. The subsequent ovulatory cycle was of normal length in most cases as compared with short 8- to 12-day cycles observed in spontaneously ovulating cows. We conclude that Kp treatment can induce ovulation in postpartum dairy cows, with ensuing oestrous cycles of normal length, if administered when a mature dominant follicle is present in the ovaries. Lay summary Cow fertility is important for efficient, profitable dairy farming. Cows that take too long after calving to become fertile are problematic. We tested a synthetically made, long-acting hormone called kisspeptin (Kp) to advance the time that cows become fertile after calving. Twenty-four dairy cows that had been calved for 3-4 weeks were used. One group of eight cows received an injection of Kp at the morning milking, another eight cows received Kp at both the morning and afternoon milking, while the last group of eight cows served as untreated controls. Kp treatment caused a desirable hormone response from the cows' brain. Normal oestrous cycles resulted, but only when a mature follicle was present in the ovary. Further study is required to analyse whether the use of a long-acting Kp drug could be used as an effective treatment for stimulating dairy cows to become more fertile after calving.

Dietary transitions, such as eliminating meat consumption, have been proposed as one way to reduce the climate impact of the global and regional food systems. However, it should be ensured that replacement diets are, indeed, nutritious and that climate benefits are accurately accounted for. This study uses New Zealand food consumption as a case study for exploring the cumulative climate impact of adopting the national dietary guidelines and the substitution of meat from hypothetical diets. The new GWP* metric is used as it was designed to better reflect the climate impacts of the release of methane than the de facto standard 100-year Global Warming Potential metric (GWP100). A transition at age 25 to the hypothetical dietary guideline diet reduces cumulative warming associated with diet by 7 to 9% at the 100th year compared with consuming the average New Zealand diet. The reduction in diet-related cumulative warming from the transition to a hypothetical meat-substituted diet varied between 12% and 15%. This is equivalent to reducing an average individual's lifetime warming contribution by 2 to 4%. General improvements are achieved for nutrient intakes by adopting the dietary guidelines compared with the average New Zealand diet; however, the substitution of meat items results in characteristic nutrient differences, and these differences must be considered alongside changes in emission profiles.

The authors would like to make the following corrections about the published paper [...]

There have been several analyses of the economics of pastoral dairy farm systems in New Zealand using real farm data, as well as several relevant international studies. However, these analyses have often used a dataset with a limited number of years that do not reflect long-term exposure to volatility, or do not allow for regional differences, and often focus on imported feed without due attention to other important characteristics of profitable farms. Several prior analyses have failed to consider the importance of a business that is resilient to major risks. We re-examined the relevance of their conclusions for New Zealand dairy systems against 12 years of DairyBase data, focussing on two major regions, deriving key insights on relevant strategic choices for profitable and resilient businesses at a farm and, by extension, industry level. Within years and regions, the top quartile of observations was identified, on the basis of ranking by operating return on assets, as a proxy for farms achieving their potential, and compared with the remaining observations. Within geographical region, the greater profitability of the top quartile was associated with greater pasture and crop eaten, greater stocking rate and production per cow, and lesser operating expenses per hectare and per kilogram milksolids (MS), defined as fat plus protein. However, greater profitability was not associated with greater use of imported feed. Linear regression was used to determine that increases in total operating expenses were associated with increases in the costs of imported feed (including winter grazing and silage made on farm). On average, for every NZ$1 spent on imported feed, total costs increased by NZ$1.66 and NZ$1.53 for the Waikato and Canterbury–Marlborough regions, respectively. This is consistent with the international literature for temperate grazing systems and is likely the reason why profitability was not greater even if above-average responses to supplement were achieved on farm. Indeed, greater use of imported feed was positively associated with operating expenses per kilogram of MS, implying that the marginal cost of additional MS was greater than the cost of the base milk, and often higher than the value of the milk produced. If gross farm revenue per kilogram MS (which is largely made up of the milk price, with a lesser contribution from livestock sales) was greater than NZ$7.50 (which it was the case in only 3 of the past 12 years), farms could generate higher profit from more imported feed use; however, the reverse was true at lower milk prices. When milk prices are low, (i.e. gross farm revenue is less than NZ$6.50/kg MS, which occurred in half of the past 12 years), farmers are often under cashflow pressure. Therefore, farm systems that are less reliant on imported feed provide a better chance for farmers to meet financial commitments, although they fail to maximise profitability when the milk price is high (e.g. >NZ$7.50/kg MS). In conclusion, maximising pasture harvested, and minimising reliance on supplementary feed, and effective cost control (minimising expenditure) are the key factors that lead to profitable businesses that are also resilient to the low milk prices that occur in volatile markets.

Extraordinary advances are occurring in biomedical science that may revolutionise how we approach health and disease. Many have applications in the dairy industry. We have described one particular area of extracellular vesicles that have already proven to be of interest in diagnostics and prognostics for fertility and assessment of ‘transition’ cows (i.e. evaluation of the problems related to the risk of clinical diseases in dairy cows, such as mastitis and milk fever, during transition period). The addition of measurements of circulating RNA and DNA may prove of value in identifying dairy cows with higher risks of clinical diseases and potentially poor fertility. We describe the exciting opportunity provided by the possibility of generating exosomes to order as therapeutic agents to potentially enhance fertility. The even more radical concept of using exosomes to deliver a CRISPR-linked gene editing function is presented. Undoubtedly, the use of biomedical advances to assist the dairy industry is an obvious and practical approach that has significant merit.

Abstract Due to the high incidence of diseases and high culling rate, the peripartum is a period of significant stress, and its management sets the stage for the health, future productivity and reproductive performance of the herd, hence making it the perfect target to improve the industry sustainability as a whole. Looking at grazing systems, using a nutrigenomic approach, we determined that, contrary to historic industry standards, cows should be slightly underfed (~75% of their energy requirements) in the three weeks before parturition to prime their metabolism and ensure an optimal transition. Farmer will be able to preserve pasture area in a period of low forage production, reaching the spring (calving season) with abundant and high quality pasture. While looking at confined systems we tested the supplementation efficacy of immunomodulatory nutrients. The greater feed intake and greater milk production performances (both in milk quantity, and quality), coupled with lower incidence of disease, underscore a smoother transition to lactation when cows are supplemented with rumen-protected methionine. Moreover, methionine induced greater blood neutrophil phagocytosis and oxidative burst postpartum, and optimized the response to LPS, by mitigating the hyper-response of the immune cells. Supplementation of cows undergoing a mastitic challenge with a S. cerevisiae fermentation product improved indexes of mammary gland health and pathogen resistance. Pathway analysis highlighted physiological mechanisms enhancing local immune cell trafficking, antimicrobial activity, and inflammation control in the mammary gland of supplemented animals, compared to control cows that showed signs of tissue damage and loss of epithelial integrity. Looking at calf nutrition, enhancing nutrient intake in the pre-weaning period stimulates mammary development, increasing their future productivity potential. Nutritional improvement of animal health shows great potential as a rapidly applicable strategy to improve animal welfare, allowing them to express their full genetic potential.

During the peripartum period, dairy cows often have signs of inflammation. Various stresses, including infectious and metabolic diseases, have been discussed as causative for this inflammation. In this study, expression profiles for 17 immune markers were measured in whole blood preparations from 78 dairy cows over a time frame starting 1 wk before calving to 4 wk after calving. Additionally, the effects of far-off and close-up feeding on immune function of dairy cows during the peripartum period were investigated. Cows were assigned to 1 of 2 feeding levels in late lactation to achieve a low and high BCS at the time of dry-off (approximately 4.25 and 5.0 on a 10-point scale). Following dry-off, both herds were managed to achieve a BCS of 5.0 one month before calving; this involved controlled feeding (i.e., maintenance) and over-feeding of ME during the far-off dry period. Within each far-off feeding-level treatment, cows were offered 65, 90, or 120% of their precalving ME requirements for 3 wk precalving in a 2 × 3 factorial arrangement. Analysis of gene expression profiles from blood cells revealed effects of time indicating that the transition cow's immune system counteracts the peripartum inflammation, whereas later postcalving it becomes activated to provide protection against postpartum infections. Far-off feeding affected (P < 0.05) the expression of 2 of the investigated genes at calving. Interleukin-6 (IL-6) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) expression in unstimulated, peripheral leukocytes were lower (P < 0.05) in animals from the Far-Off_Over-fed group compared with the Far-Off_Control-fed group. Close-up feeding had several effects on gene expression, indicating that immune function in Feed120 animals was distinct from the Feed90 and Feed65. In conclusion, feeding management precalving becomes an important intervention to ensure immunocompetence at and after calving.

Abnormalities in endometrial function contribute to poor fertility and reproductive failure. Exosomes are small lipid vesicles that contain transferable bioactive substances; they participate in intercellular signaling and may have critical roles in reproductive mechanisms, including endometrial remodeling in preparation for pregnancy. In this study, we evaluated the effects of exosomes from heifers with high and low genetic merit for fertility on inflammatory mediator expression by bovine endometrial epithelial and stromal cell lines. Co-incubation of exosomes from low, compared with high, fertility heifers upregulated the gene expression of pro-inflammatory IL1A and IL8 (CXCL8) but downregulated IL4 gene expression in epithelial cells. In contrast, stromal cells co-incubated with exosomes from low, compared with high, fertility heifers downregulated the gene expression of CXCL9, CXCL10, and CX3CL1. Our findings demonstrated that circulating exosomes from high fertility heifers did not alter endometrial inflammatory mediator gene expression. In contrast, circulating exosomes from low fertility heifers enhanced endometrial expression of inflammatory mediators, which may contribute to aberrant inflammation, leading to a reduced fertility in low fertility heifers. However, an in-depth investigation is required to elucidate the role of exosomes in regulating endometrial remodeling events required for enhanced reproductive performance and fertility in dairy cows.

Disease susceptibility of dairy cows is greatest during the transition from pregnancy to lactation. Circulating exosomes may provide biomarkers to detect at-risk cows to enhance health and productivity. From 490 cows, animals at high- (n = 20) or low-risk (n = 20) of transition-related diseases were identified using plasma non-esterified fatty acid and β-hydroxybutyrate concentrations and liver triacylglyceride concentrations during the two weeks post-calving. We isolated circulating exosomes from plasma of dairy cows at low-risk (LR-EXO) and high-risk (HR-EXO), and analyzed their proteome profiles to determine markers for metabolic dysfunction. We evaluated the effects of these exosomes on eicosanoid pathway expression by bovine endometrial stromal (bCSC) and epithelial (bEEL) cells. HR-EXO had significantly lower yield of circulating exosomes compared with LR-EXO, and unique proteins were identified in HR-EXO and LR-EXO. Exposure to LR-EXO or HR-EXO differentially regulated eicosanoid gene expression and production in bCSC and bEEL cells. In bCSC, LR-EXO exposure increased PGE 2 and PGD 2 production, whereas HR-EXO exposure increased PTGS2 gene expression. In bEEL, HR-EXO exposure caused a decrease in PGE 2, PGF 2α , PGD 2 , PGFM and TXB 2 production. The unique presence of serpin A3-7, coiled-coil domain containing 88A and inhibin/activin β A chain in HR-EXO, indicates potential biomarkers for cows at-risk for metabolic diseases. Our results are in line with the health status of the cow indicating a potential diagnostic role for exosomes in enhancing cows’ health and fertility.

The current study evaluated exosomes isolated from plasma of heifers bred to have high or low fertility through developing extreme diversity in fertility breeding values, however, key animal traits (e.g., body weight, milk production, and percentage of North American genetics) remained similar between the 2 groups. The exosomes were isolated by a combined ultracentrifugation and size exclusion chromatography approach and characterized by their size distribution (nanoparticle tracking analysis), morphology (transmission electron microscopy), and presence of exosomal markers (immunoblotting). In addition, a targeted mass spectrometry approach was used to confirm the presence of 2 exosomal markers, tumor susceptibility gene 101 and flotillin 1. The number of exosomes from plasma of high fertility heifers was greater compared with low fertility heifers. Interestingly, the exosomal proteomic profile, evaluated using mass spectrometry, identified 89 and 116 proteins in the high and low fertility heifers respectively, of which 4 and 31 were unique, respectively. These include proteins associated with specific biological processes and molecular functions of fertility. Most notably, the tetratricopeptide repeat protein 41-related, glycodelin, and kelch-like protein 8 were identified in plasma exosomes unique to the low fertility heifers. These proteins are suggested to play a role in reproduction; however, the role of these proteins in dairy cow reproduction remains to be elucidated. Their identification underscores the potential for proteins within exosomes to provide information on the fertility status and physiological condition of the cow. This may potentially lead to the development of prognostic tools and interventions to improving dairy cow fertility.

Dairy farmers in pastoral systems face a number of risks, including price volatility. Farm data was used to understand how to be profitable and able to withstand these risks. We concluded that maximising pasture harvested, and minimising reliance on supplementary feed, and effective cost control (minimising expenditure) are the key factors that lead to profitable businesses which are also resilient to the low milk prices that occur in a volatile market.

Precalving feeding level affects dry matter intake, postcalving energy balance, the risk of hepatic lipidosis and metabolic disease, and gene expression in liver and adipose tissue. These coincide with a higher risk of disease postpartum and, very likely, a failure to reach optimum production as well as reproductive targets. Current interpretation of the available evidence suggest that metabolic stressors affect the immune system of transition dairy cows and lead to reduced immunocompetence. The objective of the current study was to investigate the effect of precalving body condition score (BCS) and level of feeding on immunocompetence during the peripartum period. Twenty-three weeks before calving, 78 cows were allocated randomly to 1 of 6 treatment groups (n=13) in a 2 × 3 factorial arrangement: 2 precalving BCS categories (4.0 and 5.0, based on a 10-point scale) and 3 levels of energy intake during the 3 wk preceding calving (75, 100, and 125% of estimated requirements). Blood was sampled precalving and at 1, 2 and 4 wk after calving. Cells were analyzed by flow cytometry and quantitative real-time PCR. The numbers of T helper lymphocytes (CD4+), cytotoxic T lymphocytes (CD8+), natural killer cells (CD335+), and γδ T lymphocytes (WC1+) as well as their activation status [IL-2 receptor (CD25)+ cells] were highly variable between animals, but there was no evident effect of BCS, feeding level, or time. All groups presented with an increase in expression of cytokines in unstimulated blood cells in the week after calving, although this was significant only for IFNG in the BCS 4.0 group. Analysis of in vitro-stimulated cells allowed 2 general observations: (1) cows with high energy intake precalving (125%) had increased cytokine expression precalving, and (2) all cows had increased cytokine expression in the week after calving. The present study provides evidence that prepartum feed management can affect immunocompetence during the transition period. Considering the current results, optimally conditioned animals might benefit from a restricted precalving diet, whereas underconditioned cows can be fed to requirements.

Postpartum uterine inflammation (endometritis) in the dairy cow is associated with lower fertility at both the time of infection and after the inflammation has resolved. We hypothesized that aberrant DNA methylation may be involved in the subfertility associated with uterine inflammation. The objective of this study was to characterize genome-wide DNA methylation and gene expression in the endometrium of dairy cows with subclinical endometritis (SCE). Endometrial tissues were obtained at 29 days postpartum (n = 12), and microarrays were used to characterize transcription and DNA methylation. Analyses revealed 1,856 probes differentially expressed in animals with SCE (n = 6) compared with controls (CON, n = 6, P < 0.05, Storey Multiple testing correction) and 2,976 probes with significant correlation between gene expression and bacteriology score. No significant associations among DNA methylation and gene expression were detected. Analysis of transcription data using the Database for Annotation, Visualization, and Integrated Discovery and Gene Set Enrichment Analysis identified several pathways and processes enriched in SCE cows, with the majority related to the immune response. Furthermore, the top ontology terms enriched in genes that had expression data correlated to bacteriology score were: Defense response, inflammatory response, and innate immune response. Gene expression profiles in cows with subclinical endometritis in this study indicate that the immune response is activated, potentially resulting in a local proinflammatory environment in the uterus. If this period of inflammation is prolonged it could result in tissue damage or failure to complete involution of the uterus, which may create a suboptimal environment for future pregnancy.

Animal Production Science (continuing Australian Journal of Experimental Agriculture) publishes original research into applied agriculture including animal production, animal-plant interactions, pasture and fodder crops, field crops, extension and horticulture

Peripartum, and especially during the transition period, dairy cows undergo dramatic physiological changes. These coincide with an increased risk of disease during the first 2 wk after calving and have been linked to dairy cows failing to achieve production as well as reproductive targets. Previous evidence suggests that these physiological changes affect the immune sys-tem and that transition dairy cows experience some form of reduced immunocompetence. However, almost all of these studies were undertaken in high-production, housed dairy cows. Grazing cows have much lower levels of production and this study aimed to provide clarity whether or not the dysfunctional attributes of the peripartum immune system reported in high production housed cows are evident in these animals. Therefore, cell culture techniques, flow cytometry, and quantitative PCR were applied to analyze the cellular composition of peripheral blood mononuclear cells from transition dairy cows as well as the performance of these cells in an in vitro assay. First, a combination of in vitro stimulation and quantitative PCR for cytokines was validated as a quantifiable immunocompetence assay in 29 cattle and a correlation of quantitative PCR and ELISA demonstrated. Second, the relative number of T helper cells, cytotoxic T cells, B cells, γδ T cells, natural killer cells, and monocytes in peripheral blood was measured, of which B cells and natural killer cells increased in number postcalving (n = 29) compared with precalving. Third, following in vitro stimulation cytokine profiles indicated decreased expression of IFNγ, tumor necrosis factor, and IL-17 and increased expression of IL-10 wk 1 after calving, which later all returned to precalving values (n = 39). Additionally, treatment of transition cows with a nonsteroidal anti-inflammatory drug (i.e., carprofen) administered on d 1, 3, and 5 postcalving (n = 19; untreated control n = 20) did not affect the cytokine expression at any time point. In conclusion, an immunocompetence assay has been developed that highlights a characteristic expression pattern for IFNγ, tumor necrosis factor, IL-17, and IL-10 that reflects a state of reduced immunocom-petence in moderate-yielding pasture-based transition cows after calving, which is similar to that described for higher-yielding housed cows.