Effect of altering milking interval when milking 3 times in 2 days on milk and component yields in pasture-based dairy systems.

Dairy farmers face challenges attracting and retaining staff, partly due to the difficulty meeting the desires of the modern workforce. These include flexible work hours and regular time off. The task of milking fundamentally affects the ability of dairy farmers to meet these desires. Milking contributes to a large proportion of the hours spent working on dairy farms. The number of milkings (milking frequency) and their timing (milking interval) within a day influence the number of hours spent milking and what time in the day they occur. Milking 3 times in 2 d (3-in-2) reduces the amount of time spent milking compared with milking twice a day (TAD), without reducing milk yield as much as milking once a day (OAD). However, long intervals between 3-in-2 milkings can still lead to a long workday if farmers are expected to work between milkings. The objective of this study was to determine the effect of milking interval within a 3-in-2 milking frequency on milk yield and composition at 2 stages of lactation and compare these with OAD and TAD milking. Cows (n = 200) were milked in 5 groups of 40 at 3 intervals of 3-in-2: 8-20-20 h, 10-19-19 h, and 12-18-18 h, along with 24 h (OAD), and 10 and 14 h (TAD), for 6 wk at early lactation (mean 24 d in milk ± 7 d, SD) and again at mid lactation (mean 136 d in milk ± 18 d). Milk yields were recorded at each milking and milk samples collected weekly to determine composition. At both early and mid lactation there were no significant differences in milk, fat, protein, or lactose yields between the three 3-in-2 intervals. Cows milked 3-in-2 produced 8% less milk than cows milked TAD and 14% more than cows milked OAD, with smaller differences observed at mid lactation between TAD and 3-in-2. For a 3-in-2 milking frequency, a shorter milking interval can be implemented on the days when cows are milked twice. This may allow farmers to shorten the working day when using 3-in-2, without compromising milk or component yields.

Animal- and herd-level factors associated with onset of puberty in grazing dairy heifers.

AIMS: To explore animal- and herd-level risk factors influencing age at puberty in predominantly Holstein-Friesian dairy heifers managed in seasonal, pasture-based systems. METHODS: Heifers born in spring 2018 (n = 5,010) from 54 commercial dairy herds in New Zealand were visited on three occasions when the mean heifer age, within herd, was 10 (visit 1; V1), 11 (V2) and 12 (V3) months old. Blood samples were collected on each visit and liveweight, stature and anogenital distance (AGD) were measured at V2. Heifers were defined as having reached puberty at the first visit where blood progesterone was elevated (≥ 1 ng/mL). Animal-level response variables included pubertal status by V1, V2 and V3, and age at puberty (or age at V3 plus 31 days for those that had not attained puberty by V3). To explore herd-level management factors, farmers answered a questionnaire relating to animal location, land type, health, feeding, and management between weaning and mating. A partial least squares regression was undertaken to identify herd-level factors associated with the greatest influence on puberty rate within herd. RESULTS: The mean age at puberty was 352 (SD 34.9) days. Heavier animals at a greater proportion of expected mature liveweight based on their breeding value for liveweight, or animals with a higher breed proportion of Jersey and lower breed proportion of Holstein, were associated with earlier puberty. Herd puberty rates varied widely among enrolled herds, and averaged 20%, 39% and 56% by V1, V2 and V3, respectively. Liveweight, followed by breed and land type, had the greatest influence on the herd puberty rate. Heifer herds with a greater mean liveweight (absolute and proportion of expected mature weight) or greater Jersey proportion had more animals that reached puberty at any visit, whereas herds located on steep land or with greater Holstein breed proportions had lower puberty rates. Management-related factors such as vaccinations, provision of feed supplements, and weighing frequency were also herd-level risk factors of puberty but had less influence. CONCLUSIONS AND CLINICAL RELEVANCE: This study highlights the importance of having well-grown heifers for increasing the chances of earlier puberty onset and the effect of breed and youngstock management to achieve growth targets. These outcomes have important implications for the optimal management of heifers to achieve puberty before their maiden breeding and for the timing of measurements to potentially incorporate a puberty trait in genetic evaluations.

Improving parlor efficiency in block calving pasture-based dairy systems through the application of a fixed milking time determined by daily milk yield and milking frequency.

Adjusting end-of-milking criteria, in particular applying a maximum milking time determined by expected milk yield at an individual milking session, is one strategy to optimize parlor efficiency. However, this strategy can be difficult to apply practically on farm due to large differences in session milk yield, driven by milking interval, which affects milking routines and can be limited by in-parlor technology. The objective of this study was to test the hypothesis that a single fixed milking time (duration) could be applied at all milking sessions without compromising milk production or udder health for a range of milking intervals. To test the hypothesis, 4 experimental herds were established: (1) herd milked twice a day (TAD) using a 10- and 14-h interval, (2) herd milked TAD using an 8- and 16-h interval, (3) herd milked 3 times in 2 d using a 10-19-19-h interval, and (4) herd milked once a day (OAD). Herds consisted of 40 cows each, and were established for two 6-wk experimental periods, one in peak lactation and the other in mid-late lactation. Within each herd, half the cows had an end-of-milking criterion of 0.35 kg/min (Flow), and the other half had milking ended after a fixed period of time (FixedT) based on the average milking session yield, the daily milk yield divided by average number of milkings per day, irrespective of milking interval. We found no differences in daily milk yield between end-of-milking criteria due to residual milk from one milking likely increasing the proportion of milk in the udder cistern at the next milking session for the FixedT treatment. However, fat yield was compromised when the percentage of the herd with a truncated milking exceeded an estimated 33% at a milking session, which occurred in the TAD 8-16 herd due to the divergence from the average milking interval (in the case of TAD, 12-12 h). Applying a fixed milking time had no detrimental effects on udder health, except in the OAD herd in mid-late lactation, which had both a higher cell count and new intramammary infection rate. This warrants further investigation, although the majority of cultured bacteria were coagulase-negative staphylococci (CNS). Consequently, we conclude that, in general, with appropriate monitoring (e.g., weekly inspection) to ensure the proportion of the herd with truncated milkings does not exceed 33%, farmers in pasture-based dairy systems can use a fixed milking time to improve parlor efficiency.

Adaptive immune response ranking is associated with reproductive phenotypes in grazing dairy cows divergent in genetic merit for fertility traits.

A strong adaptive immune response has been reported to have positive effects on fertility; therefore, we investigated antibody- and cell-mediated adaptive immune responses (AMIR and CMIR, respectively) and their associations with reproductive phenotypes using a population of animals that differed in their estimated genetic merit for fertility traits (fertility breeding value; FertBV). Holstein-Friesian heifers (n = 528) grazed on pasture in 4 herds based on age. These herds included 277 heifers of positive (POS) FertBV and 251 of negative (NEG) FertBV. The adaptive immune response (IR) was evaluated before puberty at 7.5 mo of age and used to rank animals as high, average, or low for AMIR, CMIR, and overall IR (combined CMIR and AMIR). The animals were studied from 12 wk of age through to the end of their second lactation to measure growth, puberty, and timing and success of fertility phenotypes, including those related to ovulation and pregnancy. Initial analysis indicated no difference in fertility outcomes between cows ranked as high or average for AMIR (n = 55, n = 407, respectively), CMIR (n = 87, n = 354, respectively), and IR (n = 29, n = 470, respectively), so these groups were pooled as HiAv-IR. Proportions of heifers of POS FertBV were similar within HiAv and low categories across AMIR (0.52 and 0.58, respectively), CMIR (0.51 and 0.59, respectively), and IR (0.53 and 0.48, respectively). Heifers with HiAv-IR had a greater average daily weight gain from 13 to 52 wk of age (661 g, 95% confidence interval 652, 669 vs. 619 g, 95% confidence interval 591, 647) and tended to be younger at puberty (371 d, 95% confidence interval 366, 377 vs. 385 d, 95% confidence interval 369, 401) than low-IR heifers. Low-CMIR cows of a NEG FertBV had a >40 d longer calving to first ovulation interval during their first lactation compared with HiAv-CMIR NEG FertBV cows. Low-CMIR cows also had decreased pregnancy rates at both 3 wk (25% ± 7% vs. 42% ± 3%; least squares means ± standard error) and 6 wk (33% ± 7% vs. 54% ± 3%; least squares means ± standard error) into the seasonal breeding period during their first lactation, compared with HiAv-CMIR cows. In summary, although the number of POS and NEG FertBV cows was similar in each immune group; interaction effects between FertBV and immune ranking on reproductive phenotypes are evident when cows were ranked by the overall IR. There were also associations between dairy cows' CMIR ranking and ability to return to estrus and become pregnant early in the breeding period, which will have benefits in seasonal breeding systems.

Associations between peripartum lying and activity behaviour and blood non-esterified fatty acids and ß-hydroxybutyrate in grazing dairy cows.

During early lactation, most dairy cows experience negative energy balance (NEB). Failure to cope with this NEB, however, can place cows at greater risk of developing metabolic disease. Our objective was to characterise, retrospectively, lying behaviour and activity of grazing dairy cows grouped according to blood non-esterified fatty acids (NEFAs) and ß-hydroxybutyrate (BHB) as indicators of postpartum metabolic state. Blood was sampled weekly for up to 4 weeks precalving, on the day of calving (day 0), daily between 1 and 4 days postcalving, and then at least weekly between week 1 and week 5 postcalving for analysis of plasma NEFAs and BHB concentrations. Two hundred and forty-four multiparous Holstein-Friesian and Holstein-Friesian × Jersey cows were classified into one of three metabolic status groups based on maximum blood NEFAs and BHB concentrations during week 1 and 2 postcalving. A cow was classified as having either: (1) low NEFAs and low BHB (Lo-Lo; n = 78), when all blood samples were <1.0 mmol/L for NEFAs and ≤1.0 mmol/L for BHB during the first 2 weeks postcalving; (2) high NEFAs and low BHB (Hi-Lo; n = 134), when blood NEFAs were ≥1.0 mmol/L and blood BHB was ≤1.0 mmol/L at the same sampling time point during the first 2 weeks postcalving; or (3) high NEFAs and high BHB (Hi-Hi; n = 32), when blood NEFAs were ≥1.0 mmol/L and blood BHB was ≥1.2 mmol/L at the same sampling time point during the first 2 weeks postcalving. Accelerometers (IceTag or IceQube devices; IceRobotics Ltd.) were used to monitor lying and activity behaviours peripartum (-21 to +35 days relative to calving). Changes in lying behaviour and activity occurred before the mean day that cows were classified Hi-Hi and Hi-Lo (2.2 and 3.5 d postcalving, respectively). Up to 3 weeks preceding calving, Hi-Hi cows were more active, had fewer daily lying bouts (LBs), and spent less time lying than Lo-Lo cows. In addition, Hi-Hi cows had fewer daily LBs and were less active up to 4 weeks postcalving than Lo-Lo cows, but these differences were biologically small. Groups of grazing cows classified as experiencing a more severe metabolic challenge behave differently up to 3 weeks precalving than their herdmates with lower blood NEFAs and BHB postcalving. These altered behaviours may allow identification of individual cows at risk of a metabolic challenge, but further research is required.

Reducing milking frequency from twice each day to three times each two days affected protein but not fat yield in a pasture-based dairy system.

Milking 3 times in 2 d (3-in-2) could enhance the attractiveness of the dairy workplace relative to twice-a-day milking (TAD) by reducing labor requirements for milking and increasing workforce flexibility. The objective of this study was to quantify the farm system interactions associated with milking 3-in-2 at 3 stages of lactation, with the aim of providing guidance to pasture-based dairy farmers and advisors on the likely consequences of adopting 3-in-2 milking on farm productivity and business performance. Seventy-nine multiparous and 37 primiparous cows were randomly allocated to 4 experimental farms stocked at 3.5 cows/ha. One herd was milked TAD for the whole lactation (August 2019 to May 2020), with the remaining 3 milked 3-in-2 for either the whole lactation, after December 1 when cows were an average of 101 d in milk, or after March 1 when days in milk averaged 189 d. Milking intervals over 48 h were 10-14-10-14 h for TAD and 12-18-18 h for 3-in-2. Animal, pasture, and farm system data were analyzed by linear regression, with the dependent variable being the annualized value of the performance metric of interest, and the number of days in the lactation milked 3-in-2 as the independent variable. For the proportion of the season milked 3-in-2, there was a significant effect on milk (-11%), protein (-8%), and lactose (-12%) yield per cow per year, but no effect of fat. Additionally, there was a positive effect (+6%) on body condition score before dry-off and the energy required for liveweight change (+26%), and a negative effect on the energy required for walking (-30%). There were no differences in estimated feed eaten, or pasture herbage accumulation, composition, or quality. Therefore, pasture management and feed allocation under 3-in-2 should be similar to TAD. On commercial farms, the degree to which reduced milk income can be offset by lower costs will be highly farm-specific, but opportunities for savings were identified in the results. The short walking distances on the research farm and potential to improve farm management using the time saved from fewer milkings suggests better production may be achieved with 3-in-2 milking on a commercial farm.

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.

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.

Estrous activity in lactating cows with divergent genetic merit for fertility traits.

This observational study aimed to determine the effect of genetic merit for fertility traits on estrous expression and estrous cycle duration in grazing dairy cows, as measured by an activity monitoring device. A secondary aim was to describe changes in expression of estrus that occur during successive estrous cycles postpartum. Neck-mounted, activity-monitoring devices (Heatime, SCR Engineers Ltd.) were fitted to nulliparous Holstein-Friesian heifers with positive (POS FertBV) or negative genetic merit for fertility traits (NEG FertBV) to capture activity data during their first and second lactations (POS FertBV: n = 242, n = 188; NEG FertBV: n = 159, n = 87 in lactation 1 and 2, respectively). An estrous event was identified when the activity change index exceeded 26 activity units (AU) for 4 h. A total of 1,254 and 892 estrous events were identified in lactation 1 and 2, respectively. Estrous duration was defined as the interval between when the threshold was first exceeded and when activity dropped below the threshold, with no new event starting within 24 h of the end of the previous event. This definition of estrus included cows in which activity crossed the threshold multiple times in a day and were classified as a single estrous event. A second measure, high activity duration, was defined as the total hours that activity exceeded the threshold. To characterize estrous activity, peak activity (above baseline) and total activity (area under the curve of activity above baseline) were measured. Compared with NEG FertBV cows, POS FertBV cows had more active, longer estrous events. In lactation 1, the POS FertBV group had a mean estrous duration and a high activity duration of 12.5 and 12.4 h compared with 11.4 and 11.3 h for the NEG FertBV group [standard error of the difference (SED) = 0.5 and 0.4 h, respectively]. This significant difference also occurred in lactation 2, with a mean estrous duration of 13.1 versus 11.8 h (SED = 0.5 h) and a high activity duration of 13.0 versus 11.8 h (SED = 0.4 h) in the POS and NEG FertBV groups, respectively. Total activity and peak activity were greater in the POS compared with the NEG FertBV group in lactation 1 (peak activity: 65.5 vs. 55.8 AU, SED = 2.4 AU; total activity: 588 vs. 494 AU, SED = 25 AU) and lactation 2 (peak activity: 72.5 vs. 61.2 AU, SED = 2.9 AU; total activity: 648 vs. 541 AU, SED = 30 AU). Estrous cycle duration did not differ between the POS and NEG FertBV groups (lactation 1: 20.4 vs. 20.6 d, SED = 0.25; lactation 2: 20.8 vs. 21.0 d, SED = 0.28). Less estrous activity of the cow was associated with the first postpartum estrus. In contrast, the number of previous estrous events did not consistently affect the duration of the subsequent estrous cycle. The outcomes of this study provide evidence that positive genetic merit for fertility traits is associated with more overt estrous expression. Selection for these traits may improve estrous expression and thus estrous detection in commercial herds.

Investigating anogenital distance and antral follicle count as novel markers of fertility within a herd of cows with positive or negative genetic merit for fertility traits.

Using early-in-life markers of reproductive characteristics may enhance the speed and success of genetic improvement in fertility. We investigated 2 phenotypes that can be measured early in life and are moderately heritable to determine their association with traditional measures of reproductive success and genetic variation under a seasonal-calving, pasture-based system. Cows were bred to be divergent in the New Zealand Fertility Breeding Value, which estimates genetic merit for fertility. Cows consisted of 2 groups with an average positive (+5%) or negative (-5%) genetic merit for fertility traits and were expected to have large diversity in reproductive outcomes. Calves were genotyped at 41 ± 3.1 d of age (mean ± SD; n = 538), and antral follicle counts (AFC) were measured when they were postpubertal heifers before their first breeding (416 ± 15 d old; 92-d range; n = 520). The anogenital distance (AGD) was measured in 478 primiparous cows of this same population 50 to 60 d after the breeding start date when they were 881 ± 25 d old (145-d range). The AGD was shorter in animals with a positive genetic merit for fertility traits (based on parent averages). An indicator of herd reproductive success in a seasonal-calving system (recalving by 6 wk in lactation 2) was chosen for logistic regression with cross-validation, and if significant, a cut-off was calculated that categorized animals into groups. Both linear and quadratic regression was undertaken, and the model with the greatest sensitivity for detection of nonpregnant cows used. The AGD linear model was significant with a sensitivity of 64% and a specificity of 48%. This model resulted in a cut-off of 102 mm, which we used to classify cows as short (≤102 mm) or long (>102 mm) AGD animals. Primiparous cows with a short AGD were more likely to be pregnant within the first 3 and 6 wk of mating, and become pregnant as a primiparous cow, than those with a long AGD. The time from calving to conception was 20 d earlier in short AGD compared with long-AGD cows. None of the models tested for AFC were significant; therefore cows were categorized into 3 groups based on previous work in seasonal systems. However, associations between fertility phenotypes and AFC group were limited. Genomic regions of interest for AGD and AFC did not overlap, indicating phenotypes were genetically independent. Overall, AGD appears as a promising early marker of fertility in seasonal grazing systems.

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.

Positive genetic merit for fertility traits is associated with superior reproductive performance in pasture-based dairy cows with seasonal calving.

New Zealand's Fertility Breeding Value (FertBV) is reported as the percentage of a sire's daughters that calve in the first 42 d of the seasonal calving period and is an estimate of genetic merit for fertility for dairy cattle. Reproductive physiology, milk production, and changes in body weight and body condition score of 2 groups of cows divergent in FertBV (+5.0%: POS; -5.1%: NEG) were characterized during their first 2 lactations. Cows grazed fresh pasture and were managed in a seasonal calving system; they were bred by artificial insemination on observed estrus for the entire breeding period (98 d in lactation 1 and 76 d in lactation 2). During lactation 1, all animals were primiparous and were randomly allocated to 1 of 2 herds, ensuring each herd was balanced for FertBV and expected calving date. During lactation 2, cows that became pregnant during lactation 1 were managed as 1 herd. Cows not inseminated in the first 42 d of the breeding season were examined for the presence of a corpus luteum and treated with an anestrus program. On average, the interval from calving to ovulation was 19 d longer in lactation 1 and 10 d longer in lactation 2 for NEG FertBV cows. The percent of cows submitted for artificial insemination after 21 d (i.e., submission rate) was 38 and 25 percentage points greater in the POS FertBV cows during lactations 1 and 2, respectively. Pregnancy rate from 42 d of breeding was 33 and 30 percentage points greater, respectively. There was no effect of FertBV on vaginal discharge score postcalving; however, POS FertBV cows had a 50% lower risk of having subclinical endometritis (polymorphonuclear leukocytes >7%) 42 d postcalving. Interactions between FertBV and month relative to calving identified that NEG FertBV cows were fatter (greater body condition score) in the month before calving, but thinner between 3 and 5 mo postcalving. There was no effect of FertBV on lactation length, estimated 270-d milk yields, or daily milk, fat, or protein yields, and only small effects on milk fat and protein percentage across the lactations. In summary, the POS FertBV cows had superior uterine health, a shorter calving to ovulation interval, a greater submission rate, and a greater pregnancy rate earlier in the breeding season when compared with the NEG FertBV cohort. Based on these results, these may be useful phenotypes to include in genetic selection indices.

Opportunities for improving the safety of dairy parlor workers.

One aspect of a successful dairy business is attracting and retaining skilled staff. The ability to do this is influenced by workplace attractiveness, one aspect of which is worker comfort and safety. Working with heavy machinery and animals makes dairy farming a dangerous occupation relative to many other sectors. Milking is the most time-consuming task on pasture-based dairy farms, and the parlor is the logical site of a number of injuries, although little is known about their causes or their relationship to parlor design and management. To investigate worker safety further, we conducted an on-farm survey on New Zealand dairy farms between December 2018 and March 2019. The survey had 3 objectives: to identify common causes of injuries relating to milking; to describe variations in milking parlor design and management practices on dairy farms; and to identify management and parlor factors that contribute to milking injuries. Analysis of accident compensation data from the state-run accident insurer for a 5-yr period from July 2013 to June 2018 was a fourth objective. Accident claims identified as related to milking through a key word search averaged NZ$252 (NZ$1 was equivalent to US$0.60 on May 19, 2020) per claim for non-severe injuries and NZ$9,334 per claim for severe injuries, for a total of NZ$1.95 million per year. A total of 136 injuries were reported on 99 of the 560 farms surveyed, and accident insurance claims were made for 53 of these injuries. The survey found that 48% of injuries occurred while workers were attaching clusters: typically soft-tissue and fracture injuries to fingers, hands, wrists, and arms. Injuries to these body parts represented the greatest cost (51% of the annual insurance cost) and identified safe cluster attachment as an area of focus for reducing milking injuries. Potential low-cost solutions include improved cluster design and a hand-arm guard. We identified a clear seasonal trend that coincided with busy periods in a seasonal block calving system, suggesting that options to reduce fatigue in spring may be beneficial; however, the number of hours worked or number of consecutive days worked were not associated with the number of milking injuries. Those aged 20-29 yr had more injuries relative to their proportion of the workforce, indicating that more training may be beneficial (e.g., setting ergonomic milking routines). Female workers were at greater risk of injury than males; this finding may have been related to the height of the animal relative to the height of the milker.

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.

Short communication: Technologies and milking practices that reduce hours of work and increase flexibility through milking efficiency in pasture-based dairy farm systems.

To attract and retain quality employees, dairy farms must be competitive with other workplaces offering more conventional hours of work. Milking requires significant labor input and influences the start and end times of the working day, affecting flexibility to suit employee needs or availability. The use of labor-saving technology and milking management strategies could help with this challenge. Previous studies have used scenario modeling in attempt to quantify the value of in-parlor technologies, however, they have relied on assumptions about the effect of the technologies on labor in the dairy. Similarly, the effect of management strategies on work patterns, such as flexible milking intervals (changing the timing of milking), has not been evaluated. The aims of this study were to (1) quantify the milking labor requirements in a range of pasture-based dairy farm systems and (2) identify practices or technologies that facilitate efficient milking. A telephone survey of 500 dairy farmers in New Zealand was conducted during April and May 2018, with questions asked about milking practices and technology use. Predictive analysis showed that at peak lactation, milking required between 17 and 24 h/wk per worker for farms milking twice a day, representing 43 to 58% of a conventional 40-h work week, depending on parlor type (herringbone or rotary), the number of clusters, and herd size. Using milking intervals of 8 and 16 h (intervals between milkings), compared with the more usual 10 and 14 h, largely avoided starting milking before 0500 h. Eight percent of herds were milked once a day, which required between 7 and 14 h/wk per worker (18-35% of a 40-h week). ANOVA showed that for metrics that related to people (labor efficiency and work routine), using automatic teat spraying had a positive effect on efficiency. Having both automatic cluster removers and drafting were associated with longer milking times in terms of throughput and row/rotation time compared with using drafting only. The results highlight considerable opportunity to reduce the number of hours those milking (employers and employees) spend in the parlor and increase staff time flexibility through milking (e.g., intervals between milkings) and parlor management (e.g., row/rotation time) and use of specific technologies. This study provides useful data for those wishing to analyze the likely value of an in-parlor automation technology or management practice for an individual situation.

Effects of far-off and close-up transition cow feeding on uterine health, postpartum anestrous interval, and reproductive outcomes in pasture-based dairy cows.

BACKGROUND: In seasonal, pasture-based, dairy production systems, cows must recover from calving and become pregnant within two to 3 months. To achieve this, the uterus must involute and ovulation must occur and continue at regular intervals. As these processes are affected by the cows' nutritional or metabolic status post-calving, the objective of this study was to evaluate the effect of cow feeding strategies on uterine health, the length of postpartum anestrous interval, and reproductive outcomes. The treatments consisted of two feeding strategies during late-lactation and early dry period (far-off period; starting 4-month pre-calving) and three close-up dry period feeding regimes (1-month pre-calving) in a 2 × 3 factorial arrangement. We randomly assigned 150 cows to one of two far-off treatments. During late lactation, the herds (n = 75 cows per herd) were either control-fed (Controlfed) or over-fed (Overfed) to achieve a low or high body condition score (4-month pre-calving; BCS; ~ 4.25 and ~ 4.75; 10-point scale) at cessation of lactation. Within each of these treatments, three feeding levels were applied during the close-up period (1-month pre-calving): ~ 65% (Feed65), ~ 90% (Feed90), or ~ 120% (Feed120) of metabolizable energy (ME) intakes relative to pre-calving requirements. RESULTS: Uterine health improved (i.e. polymorphonucleated (PMN) cells declined) with increased feeding during the close-up period for cows in the Overfed group. The reverse was evident for the Controlfed group with the greatest PMN at the highest intakes during the close-up period. The postpartum anoestrous interval (PPAI) was shorter in cows from the Overfed group when moderately fed (Feed90) during the close-up period; in comparison, the PPAI was shorter in the Controlfed group, when those cows were overfed in the close-up period (Feed120). The cows in the Overfed treatment had greater conception and pregnancy rates if cows had moderate dry matter intakes (Feed90) during the close-up period; these reproductive variables were less under excessive feed intakes (Feed120); yet, close-up dry matter intake had little effect on conception and pregnancy rates for the Controlfed group. CONCLUSIONS: The far-off feeding strategies increased early reproductive outcomes at 3 weeks of mating. Additionally, the interaction between far-off and close-up feeding strategies resulted in high six-week pregnancy rate with a slight restriction during the close-up period but only in the far-off Overfed group. Thus, our hypothesis is supported under these conditions.

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.

Transcriptomic analysis of circulating neutrophils in metabolically stressed peripartal grazing dairy cows.

The high metabolic demand during the transition into lactation places cows at greater risk of metabolic and infectious disease than at any other time in their lactation cycle. Additionally, a change occurs in the innate immune response during this period, which contributes to increased risk of disease. In the current study, we compared the transcriptomes of neutrophils from dairy cows divergent in their metabolic health post-calving. Cows (n = 5 per risk group) were selected from a parent experiment (n = 45 cows). Those with high or low concentrations of plasma nonesterified fatty acids, plasma ß-hydroxybutyrate, and liver triacylglycerol in both wk 1 and 2 were deemed to be at "high risk" (HR) or "low risk" (LR) of metabolic dysfunction, respectively. Circulating neutrophils were isolated at 3 time points during the transition period (d 0 and wk 1 and 4 post-calving), and gene expression was analyzed using RNA sequencing. Differential gene expression between the risk groups was determined using edgeR (http://bioconductor.org), and pathway analysis was conducted using Ingenuity Pathway Analysis (Ingenuity Systems, Qiagen, Valencia, CA). Statistical analysis indicated no interaction between risk and week. Therefore, the overall effect of risk was analyzed across all time points. In total, 3,500 genes were differentially expressed between the HR and LR cows (false discovery rate < 0.05). Of these, 2,897 genes were identified by Ingenuity Pathway Analysis and used for pathway analysis. Of the relevant pathways identified, neutrophils isolated from HR cows showed downregulation of genes involved in the recruitment of granulocytes, interferon signaling, and apoptosis, and upregulation of genes involved in cell survival. The results indicate that metabolically stressed cows had reduced neutrophil function during the peripartum period, highlighting a potential relationship between subclinical metabolic disease and innate immune function that suggests that metabolic health negatively affects the innate immune system and may contribute to the state of immunosuppression during the peripartum period. In this way, the metabolic stress among the HR cows may reduce their ability to combat infection during the transition period.

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.