Genome-wide association study of age at puberty and its (co)variances with fertility and stature in growing and lactating Holstein-Friesian dairy cattle.

Reproductive performance is a key determinant of cow longevity in a pasture-based, seasonal dairy system. Unfortunately, direct fertility phenotypes such as intercalving interval or pregnancy rate tend to have low heritabilities and occur relatively late in an animal's life. In contrast, age at puberty (AGEP) is a moderately heritable, early-in-life trait that may be estimated using an animal's age at first measured elevation in blood plasma progesterone (AGEP4) concentrations. Understanding the genetic architecture of AGEP4 in addition to genetic relationships between AGEP4 and fertility traits in lactating cows is important, as is its relationship with body size in the growing animal. Thus, the objectives of this research were 3-fold. First, to estimate the genetic and phenotypic (co)variances between AGEP4 and subsequent fertility during first and second lactations. Second, to quantify the associations between AGEP4 and height, length, and BW measured when animals were approximately 11 mo old (standard deviation = 0.5). Third, to identify genomic regions that are likely to be associated with variation in AGEP4. We measured AGEP4, height, length, and BW in approximately 5,000 Holstein-Friesian or Holstein-Friesian × Jersey crossbred yearling heifers across 54 pasture-based herds managed in seasonal calving farm systems. We also obtained calving rate (CR42, success or failure to calve within the first 42 d of the seasonal calving period), breeding rate (PB21, success or failure to be presented for breeding within the first 21 d of the seasonal breeding period) and pregnancy rate (PR42, success or failure to become pregnant within the first 42 d of the seasonal breeding period) phenotypes from their first and second lactations. The animals were genotyped using the Weatherby's Versa 50K SNP array (Illumina, San Diego, CA). The estimated heritabilities of AGEP4, height, length, and BW were 0.34 (90% credibility interval [CRI]: 0.30, 0.37), 0.28 (90% CRI: 0.25, 0.31), 0.21 (90% CRI: 0.18, 0.23), and 0.33 (90% CRI: 0.30, 0.36), respectively. In contrast, the heritabilities of CR42, PB21 and PR42 were all <0.05 in both first and second lactations. The genetic correlations between AGEP4 and these fertility traits were generally moderate, ranging from 0.11 to 0.60, whereas genetic correlations between AGEP4 and yearling body-conformation traits ranged from 0.02 to 0.28. Our GWAS highlighted a genomic window on chromosome 5 that was strongly associated with variation in AGEP4. We also identified 4 regions, located on chromosomes 14, 6, 1, and 11 (in order of decreasing importance), that exhibited suggestive associations with AGEP4. Our results show that AGEP4 is a reasonable predictor of estimated breeding values for fertility traits in lactating cows. Although the GWAS provided insights into genetic mechanisms underpinning AGEP4, further work is required to test genomic predictions of fertility that use this information.

Genome-wide association study of anogenital distance and its (co)variances with fertility in growing and lactating Holstein-Friesian dairy cattle.

Anogenital distance (AGD) is a moderately heritable trait that can be measured at a young age that may provide an opportunity to indirectly select for improved fertility in dairy cattle. In this study, we characterized AGD and its genetic and phenotypic relationships with a range of body stature and fertility traits. We measured AGD, shoulder height, body length, and body weight in a population of 5,010 Holstein-Friesian and Holstein-Friesian × Jersey crossbred heifers at approximately 11 mo of age (AGD1). These animals were born in 2018 across 54 seasonal calving, pasture-based dairy herds. A second measure of AGD was collected in a subset of herds (n = 17; 1,956 animals) when the animals averaged 29 mo of age (AGD2). Fertility measures included age at puberty (AGEP), then time of calving, breeding, and pregnancy during the first and second lactations. We constructed binary traits reflecting the animal's ability to calve during the first 42 d of their herd's seasonal calving period (CR42), be presented for breeding during the first 21 d of the seasonal breeding period (PB21) and become pregnant during the first 42 d of the seasonal breeding period (PR42). The posterior mean of sampled heritabilities for AGD1 was 0.23, with 90% of samples falling within a credibility interval (90% CRI) of 0.20 to 0.26, whereas the heritability of AGD2 was 0.29 (90% CRI 0.24 to 0.34). The relationship between AGD1 and AGD2 was highly positive, with a genetic correlation of 0.89 (90% CRI 0.82 to 0.94). Using a GWAS analysis of 2,460 genomic windows based on 50k genotype data, we detected a region on chromosome 20 that was highly associated with variation in AGD1, and a second region on chromosome 13 that was moderately associated with variation in AGD1. We did not detect any genomic regions associated with AGD2 which was measured in fewer animals. The genetic correlation between AGD1 and AGEP was 0.10 (90% CRI 0.00 to 0.19), whereas the genetic correlation between AGD2 and AGEP was 0.30 (90% CRI 0.15 to 0.44). The timing of calving, breeding, and pregnancy (CR42, PB21, and PR42) during first or second lactations exhibited moderate genetic relationships with AGD1 (0.19 to 0.52) and AGD2 (0.46 to 0.63). Genetic correlations between AGD and body stature traits were weak (≤0.16). We conclude that AGD is a moderately heritable trait, which may have value as an early-in-life genetic predictor for reproductive success during lactation.

Effect of nonsteroidal anti-inflammatory drugs on the inflammatory response of bovine endometrial epithelial cells in vitro.

Chronic postpartum uterine infection detrimentally affects subsequent fertility. Nonsteroidal anti-inflammatory drugs (NSAID) are used to alleviate pain and treat inflammatory conditions in transition dairy cows with varying success. To screen the efficacy of NSAID in the absence of animal experiments, we have established an in vitro model to study uterine inflammation. Inflammation was induced in cultured bovine endometrial epithelial cells by challenging cells with an inflammation cocktail: lipopolysaccharide and proinflammatory cytokines, interleukin-1ß (IL1ß) and tumor necrosis factor α (TNFα). Release of the inflammation markers, serum amyloid A (SAA) and α-1-acid glycoprotein (αAGP), was measured by ELISA. Concentration of these markers was used to indicate the effectiveness in dampening inflammation of 5 NSAID: meloxicam, flunixin meglumine, aspirin, ketoprofen, and tolfenamic acid. Three NSAID, meloxicam, flunixin meglumine, and tolfenamic acid, were successful at dampening the release of SAA and αAGP into cell-culture supernatant, and the corresponding treated cells were selected for down-stream mRNA expression analysis. Expression of 192 genes involved in regulation of inflammatory pathways were investigated using Nanostring. Of the genes investigated, 81 were above the mRNA expression-analysis threshold criteria and were included in expression analysis. All SAA genes investigated (SAA2, SAA3, M-SAA3.2) were upregulated in response to the inflammation cocktail, relative to mRNA expression in control cells; however, AGP mRNA expression was below the expression analysis threshold and was, therefore, excluded from analysis. Treatment with NSAID downregulated genes involved in regulating chemokine signaling (e.g., CXCL2, CXCR4, CXCL5, and CXCL16) and genes that regulate the eicosanoid pathway (e.g., LTA4H, PTGS2, PLA2G4A, and PTGDS). Of the 5 NSAID investigated, meloxicam, flunixin meglumine, and tolfenamic acid are recommended for further investigation into treatment of postpartum uterine inflammation. The results from this study confirm the immunomodulatory properties of the endometrial epithelium in response to inflammatory stimuli and suggest that NSAID may be beneficial in alleviating uterine inflammation.

Temporal profiles describing markers of inflammation and metabolism during the transition period of pasture-based, seasonal-calving dairy cows.

The physiology of the dairy cow while transitioning from pregnancy to lactation is complex, with multifactorial processes studied extensively for the role they play in manifestation of disease along with associated economic losses and compromised animal welfare. Manuscripts outlining associations among nutrition, production, physiology, and genetics variables and transition cow disorders are common in literature, with blood analytes that are central to energy metabolism (e.g., nonesterified fatty acids; NEFA, ß-hydroxybutyrate; BHB) often reported. Immunity and inflammation have increasingly been explored in the pathogenesis and persistence of disorders, with cytokines and acute phase proteins well documented. However, most of these studies have involved cows fed total mixed rations, which may not always reflect profiles of blood analytes and other physiological indicators of transition cow health in grazing cows consuming fresh pasture. Considering the comparatively lesser characterization of these analytes and markers in pasture-based, seasonal-calving dairy cows, we compiled a database consisting of 2,610 cow lactations that span 20 yr of transition cow research in New Zealand. Using this database, analyte profiles from approximately 28 d precalving to 35 d postcalving were identified in dairy cows with a range of genetics, milk production potentials, and pasture-based farm management systems. These profiles characterize changes in energy reserves and metabolism (NEFA, BHB, glucose, insulin, growth hormone, insulin-like growth factor-1, leptin, body condition score, body weight), liver function (globulin, aspartate aminotransferase, glutamate dehydrogenase, gamma-glutamyl transpeptidase, bilirubin, cholesterol, liver triacylglycerides), protein metabolism (albumin, total protein, albumin:globulin ratio, creatinine, urea, creatine kinase), mineral balance (calcium, magnesium, phosphate, potassium, sodium, chloride, bicarbonate), inflammation (IL-1ß, IL-6, haptoglobin, reactive oxygen species, total antioxidant capacity), and uterine health (polymorphonuclear cells, macrophage cells, vaginal discharge score). Temporal changes are generally consistent with previously characterized homeorhetic changes experienced by the dairy cow during the transition from pregnancy to lactation in both pastoral and housed systems. Some of the profiles had not previously been presented for pastoral systems, or in some cases, presented for either system. Our results indicate that moderate-yielding dairy cows undergo similar homeorhetic changes to high-yielding housed cows; however, differences in diet composition result in greater BHB concentrations than expected, based on their milk production and NEFA concentrations. In addition, most cows were able to transition to a state of higher energy requirement following calving, albeit with an increased metabolic challenge in the liver, and only a small percentage of cows were classified with severe hepatic lipidosis or severe hyperketonemia. Increases in metabolic function of the liver were accompanied by changes in indicators of the immune system and changes in mineral balance that, combined, probably reflect the innate response to the transition from gestation to lactation.

Changes in plasma electrolytes, minerals, and hepatic markers of health across the transition period in dairy cows divergent in genetic merit for fertility traits and postpartum anovulatory intervals.

Peripartum metabolism and subsequent reproductive performance of dairy cows are linked, with maladaptation over the transition period associated with poor reproductive success. A herd of seasonal calving, grazing dairy cows was established that differed in their genetic merit for fertility traits. The heifers were produced by a customized mating program to achieve a 10-percentage point divergence in the New Zealand fertility breeding value (FertBV) as follows: +5 FertBV (POS) versus -5% FertBV (NEG), while also limiting divergence in other breeding values, including body weight, body condition score, and milk production. In this study, we aimed to characterize differences in metabolic, mineral, and metabolic stress marker profiles during their first postpartum transition period as primiparous heifers and to examine if animals with longer postpartum anestrous intervals (PPAI; more than 66 d compared with less than 35 d) had greater metabolic dysfunction. Blood was sampled at -21, -14, -7, 0, 4, 7, 10, 14, 17, 21, 28, and 35 d relative to calving in 455 primiparous cows and plasma analyzed. The NEG cows had lower concentrations of both plasma nonesterified fatty acids and ß-hydroxybutyrate at d 7 compared with POS cows. Detailed temporal profiling of various metabolic, mineral, and metabolic stress markers was undertaken in a subset of cows (n = 70). Cows were selected retrospectively to create 4 groups in a 2 × 2 factorial design with either a POS or NEG FertBV and either a short (19-35 d) or long (66-131 d) PPAI. The NEG cows tended, on average, to have lower nonesterified fatty acids and ß-hydroxybutyrate concentrations compared with POS cows across the transition period. Mean body weight and body condition score was greatest in NEG cows when compared with the POS cows and an interaction with day demonstrated this only occurred precalving. They also had indications of improved liver health precalving, with higher albumin-to-globulin ratios and lower bilirubin concentrations. Concentrations of aspartate aminotransferase were lower, and the Na-to-Cl ratio was greater in cows with a long versus a short PPAI at d 28 and d 35 after calving, potentially because of cows with a short PPAI (19-35 d) returning to estrous during this time. Magnesium concentrations were lower in NEG cows with a short PPAI from d 21 onwards, indicating NEG cows may metabolically respond to estrous differently than POS cows. The NEG-long PPAI cows had greater gamma-glutamyl transferase concentrations from calving until d 28 and lower bilirubin concentrations throughout the transition period. Together, the results demonstrate significant effects of FertBV on peripartum metabolic status. However, most of the markers tested returned to reference intervals within 4 d after calving or remained within those intervals for the whole transition period, indicating relatively minor biological effects of FertBV on transition period adaptation. The profound differences in reproductive performance among the groups was not explained by underlying differences in metabolic responses during the transition period.

Response to kisspeptin and gonadotropin-releasing hormone agonist administration in Holstein-Friesian dairy heifers with positive or negative genetic merit for fertility traits.

Previous research has identified that Holstein-Friesian dairy heifers with positive (POS) genetic merit for fertility traits (FertBV) reach puberty earlier than heifers with negative (NEG) FertBV. The hypothalamus-pituitary-gonadal (HPG) axis is functional in heifers before the onset of puberty, with increased LH release evident as heifers progress toward puberty. We investigated the functionality of the HPG axis in peripubertal Holstein-Friesian dairy heifers with divergent POS or NEG FertBV, hypothesizing that the earlier puberty onset of POS heifers is associated with earlier activation of the HPG axis than in NEG heifers. In experiment 1, we tested the dose responsiveness of POS heifers to an intravenous injection of either kisspeptin [Kiss; 2, 4, or 8 µg/kg of body weight (BW); n = 3 per dose] or a GnRH agonist (buserelin; 5, 10, or 20 ng/kg of BW; n = 3 per dose). The use of these 2 agonists investigates the status of the HPG axis in both the hypothalamus (Kiss) and pituitary (buserelin) glands. Doses of 4 µg/kg BW of Kiss and 10 ng/kg BW of buserelin produced submaximal LH responses and were used in experiment 2, in which previously unused POS (n = 22) and NEG (n = 18) FertBV heifers were challenged with both agonists at 10 and 12 mo of age in a partial crossover design. Heifers were randomly allocated to treatment groups, balanced for age and BW. The LH response to buserelin was greater in POS heifers than NEG heifers at 10 mo of age, with no difference in response at 12 mo. The FSH response to buserelin and the LH and FSH responses to Kiss did not differ between the POS and NEG heifers at either age. These results indicate an association between divergent genetic merit for fertility and the LH release to buserelin at 10 mo of age, supporting the hypothesis that gonadotropin responsiveness to a GnRH agonist is more advanced in POS heifers than in NEG heifers.

Relationships between prostaglandin concentrations, a single nucleotide polymorphism in HSD17B12, and reproductive performance in dairy cows.

Prostaglandins are involved in multiple processes important for fertility, with previous work in mice highlighting a potential role for the HSD17B12 gene in prostaglandin biosynthesis. This study aimed to determine the associations among circulating prostaglandin concentrations, a missense SNP in the HSD17B12 gene predicted to disrupt protein function, and fertility traits in first-lactation Holstein-Friesian dairy cows. We used a study population of approximately 500 animals specifically bred to have either a positive (POS, +5%) or negative (NEG, -5%) genetic merit for fertility (FertBV). Genotypes of a previously identified SNP (rs109711583) in HSD17B12 were determined, with 116 animals genotyped as AA, 215 genotyped as AG, and 153 genotyped as GG. Plasma concentrations of prostaglandin E2 and the PGF2α metabolite PGFM were determined at 3 time points (12 mo of age, 4 d postpartum, and 5 wk postpartum during first lactation) in a selection of animals with AA and GG genotypes from both the POS and NEG FertBV groups (n = 33-40 in each genotype for each FertBV group). Binary reproductive traits (yes or no) examined included submission for artificial breeding in the first 3 or 6 wk of the seasonal breeding period; conception to first service; conception during the first 6 wk of the breeding period; and pregnant at the end of the breeding period. Uterine health at 6 wk after calving was examined by evaluating the percentage of polymorphonuclear leukocytes following uterine cytology and by scoring vaginal discharge based on the presence of purulent material. The 3-wk submission rate was increased in animals that carried the G allele of the missense SNP in HSD17B12, but no differences were present among genotypes for 6-wk submission rate. The trait was additive, with each increase of the G allele increasing the 3-wk submission rate by 6 to 7%. We did not observe any consistent associations between SNP alleles and circulating PGE2 concentrations; however, a complex 3-way interaction among time, fertility group, and SNP allele was present for PGFM concentrations. Plasma concentrations of PGE2 were increased approximately 40% at 5 wk postpartum in animals that were submitted for breeding within 3 or 6 wk of the start of the breeding season, and in those that conceived during the first 6 wk of breeding, compared with those that did not. Plasma concentrations of PGFM were decreased approximately 20% in those animals that conceived to their first service and tended to be decreased in animals that were pregnant at the end of the breeding period, compared with those that were not. In summary, associations were observed between the SNP in HSD17B12 and submission rate by d 21 of the breeding season, as well as between circulating prostaglandin concentrations and fertility traits, but the SNP was not consistently linked to changes in prostaglandin concentrations. Thus, the association between submission rate by d 21 of the breeding season and the SNP in HSD17B12 were unlikely driven by changes in prostaglandins.

Effect of dose rate and timing of administration of pegbovigrastim on white blood cell responses in grazing dairy cows.

Transition dairy cows experience a decline in immune function that increases the risk of peripartum disease. One strategy to improve peripartum immune function involves the use of a commercially available cytokine: bovine granulocyte-colony stimulating factor, with the addition of polyethylene glycol to increase duration of effectiveness. Treatment with Imrestor (15 mg pegbovigrastim; Elanco) one week before expected calving date (d -7) and again on the day of calving (d 0) was previously reported to increase the neutrophil number and improve neutrophil function; as a result, the incidence of clinical mastitis was reduced. We conducted 2 experiments over consecutive years to investigate the effect of a lower dose rate (half or quarter dose rate) of Imrestor in grazing dairy cattle and reduced administration frequency: one dose instead of the recommended 2. White blood cell counts were measured to determine changes in relative cell populations in response to treatment. Neutrophil function was assessed by measuring myeloperoxidase activity. Imrestor treatment increased the numbers of neutrophils, band cells, lymphocytes, and monocytes until 14 d postcalving in a dose-dependent manner; it also increased neutrophil myeloperoxidase activity. One dose of Imrestor increased white blood cell counts and myeloperoxidase activity, but the timing, degree, and duration of the response were different relative to the recommended 2 doses and were also dependent upon when Imrestor treatment was given. One dose at d -7 relative to expected calving date did not have a lasting effect postcalving, whereas one dose only on d 0 caused a delayed effect relative to cows that received 2 doses. There was no effect of Imrestor on milk yield or on blood indicators of transition cow health. A lower dose rate of Imrestor or a single dose of Imrestor on the day of calving may be sufficient to improve neutrophil function during the early postpartum in grazing dairy cows. Large-scale field studies are required to determine whether the smaller response from lower dose rates or the timing of the immunological response to drug delivery affect animal health in early lactation.

Short communication: Replacement heifer mortality from weaning until second mating in seasonal-calving, pasture-based dairy herds in New Zealand.

The primary aim of this prospective study was to determine the postnatal mortality risk of replacement dairy heifers from weaning until the start of their second mating period (∼27 mo of age) in seasonal-calving, pasture-based dairy herds. Data were analyzed from 24 farms from the Waikato (n = 15) and Canterbury (n = 9) regions of New Zealand. All animals included in the study had an identified weaning date. From this point onward, data on animals that were euthanized, died unassisted, were culled, or were sold were recorded by the farmer on the home farm or by the grazier, according to animal location, and validated using calving, mating, culling, and sold records in their herd improvement database (MINDApro LIC, Newstead, Hamilton, New Zealand). The mortality risk from weaning to the start of the second mating period was calculated by dividing the number of deaths by the total number of enrolled animals at weaning. Mortality rate was calculated by dividing the number of deaths over the study period by the total days at risk, and reported as the mortality rate per 100 cow years; this measure was also calculated as the mortality rate per farm. A total of 3,770 animals from 24 farms had data from weaning until the farm planned start of mating when animals were ∼27 mo old. The animal-level mortality incidence risk from weaning (∼13 wk of age) to the start of their second mating (∼27 mo old) was 2.7% (95% confidence interval: 2.2 to 3.3%; 102 deaths/3,770 animals). The median farm-level mortality incidence risk was 3.0%, with a range across farms from 0 to 7.9%. There was a total of 102 deaths over 2,429,362 cow days at risk, with the mean time at risk for the animals of 646 d. The animal-level mortality was 1.53 deaths (95% confidence interval 1.26 to 1.86) per 100 cow years from weaning to second mating start date. The range in farm-level mortality rate was 0 to 4.52 deaths per 100 cow years. The hazard of death did not change throughout the study period. The results from this study are difficult to compare with international studies due to differences in study timing and duration, reporting method (mortality risk vs. mortality rate), and concerns with data validation. However, to the best of our knowledge, the results from this study indicate that postweaning, postnatal mortality under the New Zealand seasonal-calving, pasture-based system is lower than most other reported studies.

Graduate Student Literature Review: Evaluating the appropriate use of wearable accelerometers in research to monitor lying behaviors of dairy cows.

Until recently, animal behavior has been studied through close and extensive observation of individual animals and has relied on subjective assessments. Wearable technologies that allow the automation of dairy cow behavior recording currently dominate the precision dairy technology market. Wearable accelerometers provide new opportunities in animal ethology using quantitative measures of dairy cow behavior. Recent research developments indicate that quantitative measures of behavior may provide new objective on-farm measures to assist producers in predicting, diagnosing, and managing disease or injury on farms and allowing producers to monitor cow comfort and estrus behavior. These recent research developments and a large increase in the availability of wearable accelerometers have led to growing interest of both researchers and producers in this technology. This review aimed to summarize the studies that have validated lying behavior derived from accelerometers and to describe the factors that should be considered when using leg-attached accelerometers and neck-worn collars to describe lying behavior (e.g., lying time and lying bouts) in dairy cows for research purposes. Specifically, we describe accelerometer technology, including the instrument properties and methods for recording motion; the raw data output from accelerometers; and methods developed for the transformation of raw data into meaningful and interpretable information. We highlight differences in validation study outcomes for researchers to consider when developing their own experimental methodology for the use of accelerometers to record lying behaviors in dairy cows. Finally, we discuss several factors that may influence the data recorded by accelerometers and highlight gaps in the literature. We conclude that researchers using accelerometers to record lying behaviors in dairy cattle should (1) select an accelerometer device that, based on device attachment and sampling rate, is appropriate to record the behavior of interest; (2) account for cow-, farm-, and management-related factors that could affect the lying behaviors recorded; (3) determine the appropriate editing criteria for the accurate interpretation of their data; (4) support their chosen method of recording, editing, and interpreting the data by referencing an appropriately designed and accurate validation study published in the literature; and (5) report, in detail, their methodology to ensure others can decipher how the data were captured and understand potential limitations of their methodology. We recommend that standardized protocols be developed for collecting, analyzing, and reporting lying behavior data recorded using wearable accelerometers for dairy cattle.

Feeding synthetic zeolite to transition dairy cows alters neutrophil gene expression.

Synthetic zeolites are used to control the availability of dietary minerals (e.g., Ca, Mg, and P) in dairy cows. Due to calcium demand increasing with lactation onset, most cows become hypocalcemic immediately postpartum, which likely contributes to poorer immune function because calcium is important for immune cell signaling. To overcome postpartum hypocalcemia, we fed transition cows synthetic zeolite A (sodium aluminosilicate) precalving and hypothesized that it would alter calcium and thus neutrophil function during the transition period. Multiparous Holstein-Friesian cows in late gestation were randomly allocated to an untreated control group (n = 10) or a treatment group in which each cow received 500 g of zeolite A daily (n = 10) for 14 d prior to the expected calving date (actual duration = 17 ± 3 d prepartum). The cows grazed pasture, and each was supplemented with 2 kg/d of maize silage (dry matter basis), with or without zeolite, until calving. Blood samples for neutrophil isolation and analysis of plasma indicators of mineral status, energy status, liver function, and inflammation were collected pretreatment (covariate; d -19); on d -14 and -7 precalving; on the day of calving (d 0); and on d 1, 4, 7, and 28 postcalving. Neutrophils were isolated and gene expression was analyzed using microfluidic gene expression arrays. Neutrophil respiratory burst was assessed using stimulation with phorbol 12-myristate 13-acetate and flow cytometry. Plasma calcium and phosphorus revealed a treatment by time interaction; cows offered zeolite had greater plasma calcium concentrations at d 0, 1, and 4 postcalving and plasma phosphorus concentrations were lower in zeolite-treated cows during the precalving period until d 1 postcalving compared with control animals. Zeolite treatment downregulated neutrophil gene expression of CXCR4 and S100A8 and tended to lower gene expression for other immune mediators (CXCR1, IFNG, S100A12, and S100A9) compared with the control. Zeolite treatment did not affect neutrophil respiratory burst or expression of the other genes investigated. Plasma concentrations of cytokine IL-6 were reduced with zeolite treatment, which was most evident immediately postcalving (d 0, 1, and 7). Overall, feeding zeolite precalving had few effects on neutrophil gene expression and function; however, the lower gene expression of neutrophil inflammatory mediators may be due to altered availability of dietary minerals prepartum and indicates that zeolite A may control inflammation during the transition period.

Associations between lying behavior and activity and hypocalcemia in grazing dairy cows during the transition period.

Hypocalcemia is a common metabolic disorder of transition dairy cows that is considered a gateway disease, increasing the risk of other health disorders and reducing cow performance. Clinical milk fever is associated with long periods of recumbency, and it is plausible that cows experiencing non-paretic hypocalcemia may spend more time lying; hence, lying behavior and activity measures may be useful in identifying at-risk cows. The objective of this study was to describe associations among blood calcium (Ca) status at calving and lying behavior and activity measures during the transition period in grazing dairy cows. Blood was sampled on the day of calving (d 0), and d 1, 2, 3, and 4 postcalving, and analyzed for total plasma Ca concentration. Twenty-four multiparous Holstein-Friesian and Holstein-Friesian × Jersey grazing dairy cows were classified, retrospectively, as clinically hypocalcemic (CLIN; blood Ca ≤ 1.4 mmol/L at 1 or more consecutive samplings within 48 h postcalving, but without parturient paresis). These cows were pair-matched (using milk production potential from their estimated breeding value for milk protein, mean body weight at wk -5 and -6 precalving, and, where possible, parity) with 24 cows classified as subclinically hypocalcemic (SUB; blood Ca > 1.4 and < 2.0 mmol/L at 2 consecutive samplings within 48 h postcalving), and 24 cows classified as normocalcemic (NORM; blood Ca ≥ 2.0 mmol/L at 3 consecutive samplings within 72 h postcalving). Lying behavior and activity were monitored using triaxial accelerometers from -21 to +35 d relative to calving. Data were summarized to calculate daily lying time (h/d), daily number of lying bouts (LB; no./d), mean LB duration (min/bout), and the number of steps taken (steps/d). On d 0, the CLIN group were less active and spent approximately 2.6 h longer lying than the SUB and NORM groups, particularly between 0200 and 1400 h. On d 0, the NORM group had fewer LB (16.3/d) than the SUB and CLIN groups (18.2 and 19.2/d, respectively). These differences in behavior were no longer detected 2 d postcalving, and no further differences were observed. The day before calving, the CLIN group spent 1.4 h longer lying down than did the SUB and NORM groups. Further, the relative change in steps from a precalving baseline period (d -14 to -7) until d 0 was positively, linearly associated with blood Ca concentration within 24 h postcalving. Future work should consider daily and temporal changes in behavior in individual cows to determine the potential for these measures to allow early detection of hypocalcemia.

A quantitative case study assessment of changes to hepatic metabolism from nonlactating grazing dairy cows consuming a large proportion of their diet as fodder beet.

Because of its high yield and the ability of cows to graze it in situ, fodder beet (FB) has become a popular crop in grazing systems, particularly for nonlactating cows. Due to its high sugar content, however, the transition to FB must be managed carefully to avoid rumen acidosis and associated metabolic dysfunction. The initial consumption of FB reduces ruminal pH; however, it is unclear whether this affects liver metabolism and results in systemic inflammation, as has been reported during subacute ruminal acidosis from high-grain diets. We used a quantitative case study approach to undertake additional measurements on a project demonstrating the effects of FB on urinary nitrogen excretion. The objective of our component, therefore, was to determine whether the inclusion of high rates of FB in the diet of nonlactating cows changed indicators of hepatic metabolism relative to a standard diet for nonlactating grazing cows. During the nonlactating period, multiparous, pregnant Holstein-Friesian cows were randomly assigned (n = 15 per treatment) to either pasture (8 kg of DM/cow per day) with corn silage (4 kg of DM/cow per day; PA) or transitioning onto an FB diet (8 kg of DM/cow per day) with pasture silage (4 kg of DM/cow per day; BT) over 14 d. Blood was sampled and the liver was biopsied during the adaptation period and after 7 d of full diet allocation. The hepatic expression of genes involved in peroxisomal oxidation was increased in cows adapting to FB, whereas the expression of genes involved in mitochondrial oxidation was increased when cows were on their full allocation of FB. These results indicate changes to fatty acid metabolism with FB consumption. Expression of 2 genes involved in the unfolded protein response was greater during the adaptation period in cows consuming FB, potentially reflecting negative effects of transitioning onto the FB diet on hepatic metabolism. Interestingly, expression of genes involved in the methionine cycle was increased in the BT cows. We hypothesize that this is a result of FB betaine absorption, although it is unclear to what extent betaine escapes ruminal degradation. While on the full diet allocation, there were lower serum concentrations of markers of hepatic stress in BT cows and no difference in expression of genes involved in oxidative stress compared with pasture-fed cows. However, there was an increase in plasma haptoglobin concentrations, indicative of an acute inflammatory response in BT cows. From this case study, we conclude that the results indicate no negative effects of the FB diet on liver metabolism and, possibly, positive effects on hepatic function. It appears, therefore, that the transition of nonlactating cows onto an FB diet can be managed to minimize the negative effects of the high sugar intake. Further research on the amount of betaine that escapes ruminal degradation in cows consuming FB would be of value to better understand whether betaine reduces liver damage in dairy cows consuming FB.

A redefinition of the modeled responses of mammary glands to once-daily milking.

Milking cows once daily is a management tool that has been implemented to improve physical and financial results of seasonal pasture-based dairy farms. The Molly cow model integrates physiology and metabolism of dairy cattle; however, milk production during short-term changes in milking frequency (e.g., 1× milking) is not well represented. The model includes a representation of variable rates of cell quiescence and death. However, the rate constants governing cell death and the return of quiescent to active cells are not affected by milking frequency. An empirical assessment of the problem was conducted, and it was hypothesized that changing the current representation of the rate of cell death in response to short-term 1× milking would more accurately represent active and quiescent cells and improve predictions of milk production. An extra senescent cell flux was added to account for cell loss during periods of 1× milking. Additional changes included a gradual decline in the rate of 1× stimulated senescence during 1× milking, and a structural change in cell cycling between active and quiescent cells during and after short-term 1× milking. Data used for parameter estimation were obtained from 5 studies where 1× milking or different feeding strategies were tested. Parameter estimates of cell loss indicated that 1× milking would affect a small proportion of quiescent cells to cause extra cell death. This added cell senescence was influenced by the length of 1× milking such that cell senescence peaked on d 1 of 1× milking and decayed from that point. The new structure in the model includes a variable rate of cell death in response to 1× milking and a gradual rate of return of quiescent cells back to the active pool in response to switching to 2× milking after short-term 1× milking. Root mean square errors, mean bias, and slope bias declined by at least 50% for predictions of energy-corrected milk yield and fat percent. The model showed quantitative agreement with production data from short-term 1× milking. The accuracy of predictions was improved and the error was reduced by implementing modifications in the model in response to changes in milking frequency.

Lying behavior and activity during the transition period of clinically healthy grazing dairy cows.

Lying behavior and activity may provide useful information for the prediction of an imminent calving and the health of transition dairy cows; however, it is important first to understand what constitutes typical lying behavior and activity because this has not been defined for grazing dairy cows during the transition period. Our objective was to describe changes in lying behavior and activity in grazing dairy cows during the transition period using varying phenotypes typical of commercial dairy herds under grazing systems. Behavior data from IceTag or IceQube (IceRobotics, Edinburgh, Scotland) triaxial accelerometers were collected for 310 cow parities from multiparous, mixed-age (mean ± standard deviation; 4.5 ± 1.65 yr), and mixed-breed [Holstein-Friesian (HF), n = 216; and HF × Jersey, n = 94] grazing dairy cows from 4 parent experiments. The IceTags or IceQubes captured lying and activity data during the transition period (-21 to 34 d relative to calving) to allow the calculation of daily lying time (h/d), daily lying bouts (LB; no./d), mean LB duration (min/bout), and the number of steps taken (steps/d). Lying behavior and activity were analyzed using a repeated measures ANOVA during 3 periods: PRE (-21 to -3 d), POST (3 to 21 d), and the day of calving (d 0). Lying time was lower on d 0 (7.25 h/d) compared with PRE and POST lying times (10.3 and 8.58 h/d, respectively), with more frequent LB on d 0 (12.9 no./d) compared with the PRE and POST daily LB (8.15 vs. 7.74 no./d). Cows took more steps POST (4,424 steps/d) compared with d 0 and PRE (4,105 and 2,289 steps/d, respectively). Regression analysis determined that daily lying time decreased substantially from -3 to 0 d (slope = -1.03 ± 0.07 h/d) and from -2 and -1 d for daily LB (slope = 5.09 ± 0.54 no./d), which may be due to the calving event itself but also reflect restlessness. Daily lying time, daily LB, LB duration, and number of steps taken were substantially altered at the time of the calving event in grazing dairy cows. Cows were more active, spent less time lying, and took more steps postcalving compared with precalving, and it appears that this behavior may largely be due to activity associated with twice daily milking. Mean lying behavior and activity measures were more highly variable across individuals than across groups. Information available via activity monitors may contribute to the improvement of individual management of transition dairy cows, and this research provides a benchmark for typical changes in behavior during the transition period in grazing systems.

Prediction of blood ß-hydroxybutyrate content and occurrence of hyperketonemia in early-lactation, pasture-grazed dairy cows using milk infrared spectra.

The objective of this study was to evaluate the ability of milk infrared spectra to predict blood ß-hydroxybutyrate (BHB) concentration for use as a management tool for cow metabolic health on pasture-grazed dairy farms and for large-scale phenotyping for genetic evaluation purposes. The study involved 542 cows (Holstein-Friesian and Holstein-Friesian × Jersey crossbreds), from 2 farms located in the Waikato and Taranaki regions of New Zealand that operated under a seasonal-calving, pasture-based dairy system. Milk infrared spectra were collected once a week during the first 5 wk of lactation. A blood "prick" sample was taken from the ventral labial vein of each cow 3 times a week for the first 5 wk of lactation. The content of BHB in blood was measured immediately using a handheld device. After outlier elimination, 1,910 spectra records and corresponding BHB measures were used for prediction model development. Partial least square regression and partial least squares discriminant analysis were used to develop prediction models for quantitative determination of blood BHB content and for identifying cows with hyperketonemia (HYK). Both quantitative and discriminant predictions were developed using the phenotypes and infrared spectra from two-thirds of the cows (randomly assigned to the calibration set) and tested using the remaining one-third (validation set). A moderate accuracy was obtained for prediction of blood BHB. The coefficient of determination (R2) of the prediction model in calibration was 0.56, with a root mean squared error of prediction of 0.28 mmol/L and a ratio of performance to deviation, calculated as the ratio of the standard deviation of the partial least squares model calibration set to the standard error of prediction, of 1.50. In the validation set, the R2 was 0.50, with root mean squared error of prediction values of 0.32 mmol/L, which resulted in a ratio of performance to deviation of 1.39. When the reference test for HYK was defined as blood concentration of BHB ≥1.2 mmol/L, discriminant models indicated that milk infrared spectra correctly classified 76% of the HYK-positive cows and 82% of the HYK-negative cows. The quantitative models were not able to provide accurate estimates, but they could differentiate between high and low BHB concentrations. Furthermore, the discriminant models allowed the classification of cows with reasonable accuracy. This study indicates that the prediction of blood BHB content or occurrence of HYK from milk spectra is possible with moderate accuracy in pasture-grazed cows and could be used during routine milk testing. Applicability of infrared spectroscopy is not likely suited for obtaining accurate BHB measurements at an individual cow level, but discriminant models might be used in the future as herd-level management tools for classification of cows that are at risk of HYK, whereas quantitative models might provide large-scale phenotypes to be used as an indicator trait for breeding cows with improved metabolic health.

Economic costs of recorded reasons for cow mortality and culling in a pasture-based dairy industry.

The objective of this study was to determine the economic costs associated with different reasons for cow culling or on-farm mortality in a pasture-based seasonal system. A bioeconomic model was developed to quantify costs associated with the different farmer-recorded reasons and timing of cow wastage. The model accounted for the parity and stage of lactation at which the cows were removed as well as the consequent effect on the replacement rate and average age structure of the herd. The costs and benefits associated with the change were quantified, including animal replacement cost, cull salvage value, milk production loss, and the profitability of altered genetic merit based on industry genetic trends for each parity. The total cost of cow wastage was estimated to be NZ$23,628/100 cows per year (NZ$1 = US$0.69) in a pasture-based system. Of this total cost, NZ$14,300/100 cows worth of removals were for nonpregnancy and unknown reasons, and another NZ$3,631/100 cows was attributed to low milk production, mastitis, and udder problems. The total cost for cow removals due to farmer-recorded biological reasons (excluding unknown, production, and management-related causes) was estimated to be NZ$13,632/100 cows per year. Of this cost, an estimated NZ$10,286/100 cows was attributed to nonpregnancy, mastitis, udder problems, calving trouble, and injury or accident. There is a strong economic case for the pasture-based dairy industries to invest in genetic, herd health, and production management research focused on reducing animal wastage due to reproductive failure, mastitis, udder problems, injuries or accidents, and calving difficulties.

Fertility and the transition dairy cow.

The transition from pregnancy to lactation (i.e. the transition period) is a time of significant metabolic challenge, with a several-fold increase in a cow's requirement for energy, protein and minerals within days of calving. A successful transition involves the initiation and coordination of changes in multiple tissues that facilitate the provision of these nutrients to the cow and, more specifically, to the mammary gland, often at the considerable expense of other tissues. Failure to coordinate the necessary changes effectively results in transition period maladaptation, which can broadly be grouped into three categories: (1) negative energy balance and metabolic diseases associated with energy metabolism; (2) immune dysfunction and inflammation; and (3) metabolic diseases associated with mineral deficiency. Because reinitiation of ovarian activity, follicle recruitment, ovulation, fertilisation and, potentially, even maternal recognition of pregnancy and implantation occur against the backdrop of this metabolic and immunological disturbance in early lactation, the role of nutrition in ensuring a smooth transition between the pregnant and lactating state is important. In this paper we integrate recent research findings with previous knowledge of the interaction between transition cow metabolism and nutrition and reproductive outcomes, and offer new insights into key elements of successful cow management to avoid transition 'maladaptation' and improve pregnancy rates.

Calf and replacement heifer mortality from birth until weaning in pasture-based dairy herds in New Zealand.

The aims of this study were to (1) estimate the perinatal (birth to 24 h) and postnatal (∼24 h to the mean weaning age of 13 wk) mortality risk in pasture-based dairy calves until weaning, and (2) identify associated risk factors in the 2015 calving season. A prospective survey of 32 seasonal calving dairy farms was undertaken. Farmers recorded (daily) the number and sex of the calves alive or dead in the paddocks where cows calved. All daily animal movements in and out of the calf rearing facilities, including death and euthanasia, and the identification of the animals (if applicable) were recorded, and a survey of the farm management practices was undertaken. Individual and farm-level risk factors for perinatal mortality were modeled separately using generalized logistic mixed models with a random effect fitted for herd. Postnatal mortality incidence risk was calculated using time at risk for each calf from 24 h of age, collapsed into weeks, and multiplying the incidence risk by the mean weaning age of the study population. Farm-level risk factors contributing to postnatal mortality in the first week of life were assessed using a multivariable logistic mixed regression model. The mean perinatal mortality risk was 5.7% (95% confidence interval 5.4 to 6.1%) with a range from 2.2 to 8.6% (18,437 calves, 30 farms). Perinatal calf mortality was greater for male relative to female calves (odds ratio 1.39; 95% confidence interval 1.22 to 1.59), calves born in the first week of the calving period in comparison to wk 2 to 11 (odds ratio 0.32 to 0.66), and those born on days with greater rainfall (odds ratio 1.01 per 1 mm increase; 95% confidence interval 1.00 to 1.02). At the farm level, perinatal mortality increased for every extra week of calving period length (odds ratio 1.12; 95% confidence interval 1.06 to 1.17). The mean postnatal mortality risk was 4.1% (95% confidence interval 3.6 to 4.6%) with a range of 0 to 11% between farms. Farm-level risk factors contributing to mortality in the first week of life included farmer-reported disease problems in calves (odds ratio 2.2; 95% confidence interval 1.62 to 2.96), or calves hand-fed colostrum in the first 12 h of life (odds ratio 1.66; 95% confidence interval 1.26 to 2.19), which was assumed to be associated with poor colostrum quality and bacterial contamination. Regional differences were also observed in both perinatal and postnatal mortality risks, indicating that weather conditions, herd size, and management variations contribute to mortality incidence. In summary, the mortality risk of perinatal calves and postnatal calves until weaning on pasture-based farms is comparable with data published from other dairying systems despite the notable management differences. Several risk factors were identified that could be managed to reduce mortality incidence of dairy calves.

Invited review: A systematic literature review and meta-analysis of mortality and culling in dairy cattle.

Dairy industries and individual farmers are concerned about mortality and culling of dairy animals. This is because the timing and fates of animals that exit dairy farms have important animal welfare and economic consequences that reflect the conditions under which they are farmed and the efficiency of their production systems. Reports from a few countries have indicated increased incidence of mortality, and occasionally culling, of dairy animals in recent decades, and these changes have been associated with intensification of production systems. Dairy industries and farmers need benchmarks for culling and mortality against which they can compare themselves, as well as improved understanding of the extent of any change and of any associated factors. We reasoned that a systematic literature review and meta-analysis of scientific articles published between 1989 and 2014 would allow us to determine whether these reports were universal, to quantify any change over time, and to investigate whether production systems or study factors were associated with culling and mortality. From 3,275 articles retrieved from databases and manual searching of cited articles, 118 articles were appraised independently by 2 assessors, and 51 articles representing 54 studies were determined to be eligible for review and meta-analysis. We estimated that both the annual incidence risk (IR) and incidence density of mortality of cows had increased significantly from 0.02 per cow and 2.32 per 100 cow-years, to 0.04 per cow and 3.75 per 100 cow-years, an increase per decade of 0.02 per cow and 1.42 per 100 cow-years, respectively. We also estimated that the annual IR of culling attributed to low production had declined significantly from 0.07 to 0.05 and that the IR of perinatal, but not neonatal, mortality had increased significantly from 0.04 to 0.06 per decade. We found no evidence of change in overall annual IR of culling of cows over time or any association between study design factors and the IR or incidence density of culling or mortality. These findings provide benchmarks for describing culling and mortality, and should encourage farmers and researchers in countries with modern dairy industries to discover and implement management strategies to reduce the animal welfare and economic costs associated with these changes.