Graduate Student Literature Review: Implications of transition cow health for reproductive function and targeted reproductive management.

Negative associations of health disorders with reproductive performance, often measured with pregnancy risk per artificial insemination (AI) or the risk of pregnancy loss, have been demonstrated extensively. Most studies investigated common clinical diseases but did not include subclinical disorders comprehensively. They often evaluated cows subjected to hormonal synchronization protocols for timed AI, limiting the ability to understand how disease may affect spontaneous reproductive function, which is essential for targeted management programs with selective hormonal intervention. It is plausible that metabolic and inflammatory disorders have short- and long-term detrimental effects on different features of reproductive function that result in or contribute to reduced fertility. These may include: 1) reestablishment of endocrine function to promote follicular growth and first ovulation postpartum, 2) corpus luteum (CL) function, 3) estrus expression, and 4) uterine environment, fertilization, and embryonic development. In this narrative literature review, we discuss insights and knowledge gaps linking health disorders with these processes of reproductive function. A growing set of observational studies with adequate internal validity suggest that these outcomes may be affected by metabolic and inflammatory disorders that are common in the early postpartum period. A better characterization of these risk factors in multi-site studies with greater external validity is warranted to develop decision-support tools to identify subgroups of cows that are more or less likely to be successful in targeted reproductive management programs.

Randomized clinical trial of ketoprofen or ceftiofur for treatment of metritis in dairy cows.

Our objectives were to compare the efficacy of ketoprofen or ceftiofur for treatment of metritis in dairy cows considering subsequent health, production, and reproduction. Cows from 2 commercial dairy farms in Ontario, Canada were examined with a Metricheck device 3 times per week from 2 to 14 d in milk (DIM). Cows with metritis (fetid vaginal discharge; n = 193) were blocked by parity and fever (rectal temperature ≥39.5°C or <39.5°C) and within each block per farm, randomly assigned to receive 3 mg/kg BW of ketoprofen (KET) or 2.2 mg/kg of ceftiofur hydrochloride (CEF), once a day for 3 d. Day of enrollment was considered study d 0. Rectal temperature and attitude were evaluated in cows with metritis on study d 0, 3, 4, 7, 10, and 13, and vaginal discharge was evaluated on study d 4, 7, 10, and 13. Body condition was scored at enrollment and 35 DIM, and serum concentration of haptoglobin was measured at d 0, 2, 4, and 7. Cows with rectal temperature ≥39.5°C or a depressed attitude on d 3 were classified as clinical failure and received treatment with ceftiofur for 3 d (KET), or 2 additional days (CEF), to a maximum of 5 d of treatment with ceftiofur. At 35 ± 3 DIM cows were examined for uterine involution by transrectal palpation, purulent vaginal discharge (PVD) by Metricheck, and endometritis by endometrial cytology. Time to onset of cyclicity was assessed by serum progesterone (P4) measurements at 28, 42, and 56 DIM. Contemporary cows from the same farms without metritis (NOMET; n = 1,043) were used for comparison. Data were analyzed with mixed linear or logistic regression or Cox's proportional hazard models, including herd as a random effect. The proportion of clinical resolution of metritis on d 3 (96% vs. 92%), of cows with fever (from d 3 to d 13 after enrollment) or fetid discharge (from d 4 to d 13 after enrollment), and the number of medical treatments (3.1 vs. 3.3) were not different between CEF and KET, respectively. Cows in KET received fewer antibiotic treatments than cows in CEF (0.3 vs. 3.1). Uterine involution, the prevalence of PVD (50% vs. 47%) and subclinical endometritis (6.6% vs. 4.3%), and the proportion of cyclic cows (82% vs. 86%) did not differ between CEF and KET. Cows in KET had greater serum haptoglobin concentration from d 2 to 7 after enrollment. The incidence of mastitis, lameness, or displaced abomasum to 60 DIM and subclinical ketosis to 21 DIM did not differ among CEF, KET, and NOMET. There were no differences in median days to first AI (CEF = 68 d; 95% CI: 65-70; KET = 69 d; 95% CI: 68-72; NOMET = 69 d; 95% CI: 68-70), and median days to pregnancy (CEF = 118 d; 95% CI: 92-145; KET = 113 d; 95% CI: 90-135; NOMET = 105 d; 95% CI: 101-109), pregnancy at first AI at 33 d after insemination (CEF = 42%; KET = 41%; NOMET = 41%), pregnancy loss after first AI (CEF = 8%; KET = 11%; NOMET = 8%), hazard of pregnancy or hazard of culling up to 300 DIM. Milk yield was not different between CEF and KET during the first 10 weeks, but lesser in KET at wk 2 and 4 and CEF at wk 2, 4, and 6 than in NOMET. In this pilot-scale study, given early detection, we did not detect differences in subsequent health, milk yield, or reproductive performance in cows with metritis initially treated for 3 d with CEF or KET. Additional, larger studies are warranted.

Effects of systemic or uterine endotoxin challenge in Holstein cows at 5 or 40 days postpartum on clinical responses, uterine and systemic inflammation, and milk yield.

Our objective was to investigate the effects of intravenous (IV) or intrauterine (IU) lipopolysaccharide (LPS) challenge at 5 or 40 d postpartum (DPP) on clinical signs, systemic and uterine inflammation, dry matter intake (DMI), and milk yield (MY). Holstein cows at 5 DPP (n = 23) or at 40 DPP (n = 24) were blocked by parity and randomly assigned to one of 3 treatments: 1) IV-LPS [0.0625 µg/kg BW (5 DPP) or 0.1 µg/kg BW (40 DPP) over 1h], 2) IU-LPS [100 µg (5 DPP) or 300 µg (40 DPP) in 20 mL saline], or 3) 20 mL saline IU (IU-SAL; same for 5 and 40 DPP). The proportion of polymorphonuclear (PMN) cells was measured by endometrial cytology at d -1, 1, 4, and 7 relative to treatment. Blood haptoglobin (Hp), serum-amyloid A (SAA), and LPS-binding protein (LBP), DMI, and MY were measured from d -1 through 7. Data were analyzed separately for each DPP group in multivariable linear regression models accounting for repeated measures. For both DPP groups, there were increases in rectal temperature, heart and respiratory rates, and a decrease in rumination rate following IV-LPS, but not following IU-LPS. At 5 DPP, endometrial PMN proportion was similar in IU-LPS and IU-SAL. Serum Hp was unaffected by LPS challenge, SAA was greater in IV-LPS from 12 to 24 h after challenge, and LBP was greater in IV-LPS from 8 to 24 h. At 40 DPP, PMN was greater in IU-LPS (37 ± 4%) than in IU-SAL (15 ± 4%) 1 d after LPS challenge. Serum Hp was greater from 24 to 72 h after challenge in IV-LPS than in the other groups, SAA was greater in IV-LPS from 6 to 48 h, and LBP was greater in IV-LPS from 8 to 24 h. At both 5 and 40 DPP, treatment did not affect DMI, but MY was lesser in IV-LPS cows at 12 and 24 h than in IU-SAL or IU-LPS. The IV-LPS challenge resulted in more pronounced changes in clinical signs and acute phase protein (APP) concentrations than IU-LPS or IU-SAL at 40 DPP, but more subtle or inconsistent changes at 5 DPP. These may be due to the different doses of LPS used at 5 and 40 DPP or possibly due to the high variation in baseline clinical signs and APP observed in all groups at 5 DPP. The IU-LPS increased uterine PMN 1 d after challenge at 40 DPP, but not at 5 DPP. At each time, IU-LPS did not produce changes in clinical signs or markers of systemic inflammation.

Progesterone profiles in postpartum dairy cows with inflammatory disorders.

The objective of this prospective cohort study was to determine if progesterone (P4) profiles differed between dairy cows with or without inflammatory disorders early postpartum. A total of 708 cows from 2 commercial herds were enrolled 3 wk before parturition and examined for clinical health disorders (retained placenta, metritis, displaced abomasum, mastitis, or lameness) until 5 wk postpartum. Serum haptoglobin (Hp) was measured in blood at 2 and 6 (±2) DIM, metritis was assessed at 4, 8, 11, and 15 DIM, and purulent vaginal discharge and endometritis (≥6% polymorphonuclear cells in endometrial cytology sampled by cytobrush) were assessed at 35 ± 3 DIM. As Hp ≥0.8 g/L or endometritis were associated with ovarian dysfunction in previous studies, cows with serum Hp ≥0.8 g/L at either time point and endometritis, regardless of clinical disease, were classified as the cohort with inflammatory disorders (INFLAM; n = 139). Clinically healthy cows without difficult calving or twin birth, with Hp <0.8 g/L at both sampling times, without endometritis, and BCS ≥3.00 (1 to 5 scale) were classified as healthy (n = 133). Cows with only one of the 2 conditions (high Hp or endometritis) were excluded. Cohorts had serum P4 measured twice weekly from 35 to 70 (±3) DIM, and the first detected luteal phase (LP) during the sampling period was defined as the interval from onset of luteal activity (P4 increase to ≥1 ng/mL) until decline of P4 to <1 ng/mL. The odds of prolonged LP (≥21 d), average LP length, peak P4, and time to P4 decline (hazard rate) were analyzed using multivariable mixed logistic, linear, or Cox proportional hazard regression models including INFLAM status, parity, sampling day (when applicable), and herd as a random effect considering the covariates of season, milk yield at first DHIA test, and DIM at onset of cyclicity or LP length (when applicable). Cows with INFLAM had greater odds of prolonged LP (LSM ± SEM; 67% vs. 37% ± 7), greater average LP length (17 vs. 15 ± 2 d), lesser P4 at d 4 (4.6 vs. 5.5 ± 0.3 ng/mL) and d 7 (6.0 vs. 7.7 ± 0.3 ng/mL) of the LP, and lesser peak P4 (6.9 vs. 8.2 ± 0.3 ng/mL) during the LP than healthy cows. Status of INFLAM was associated with time to P4 decline in multiparous but not primiparous cows; the LP of INFLAM multiparous cows was less likely to have luteolysis (P4 decline) by d 14 [adjusted hazard ratio (AHR) and 95% CI: 0.54; 0.31 to 0.94] or by d 21 (AHR: 0.32; 0.12 to 0.84) than in healthy multiparous cows. In conclusion, postpartum cows with markers of systemic inflammation at wk 1 and uterine inflammation at wk 5 had altered luteal function (prolonged LP and lower P4 concentrations) before first breeding, which is a possible pathway linking postpartum health disorders and infertility.

Evaluation of bovine colostrum replacer supplementation to improve weaning transition in Holstein dairy calves.

The objective of this randomized controlled trial was to evaluate the efficacy of supplementing bovine colostrum replacer during weaning to reduce intestinal permeability and improve gain. For this experiment, 65 calves were enrolled and housed individually until 70 d of age. Calves were fed milk replacer (150 g/L) 3 times daily with 9 L/d, 10.5 L/d, 11.25 L/d, and 12 L/d offered from d 1-7, d 8-14, d 15-21, d 22-56, respectively. Calves were weaned over 8 d from d 57-64, receiving a total of 7.8 L in 2 meals/d from d 57-60 and 3.8 L/d in one feeding from d 61-64. At d 57, calves were blocked by birth weight and randomly assigned to one of 2 treatments, equal in metabolizable energy, which were fed once daily during weaning from d 57-64: control (CON; n = 31 calves): 3.8 L milk replacer (150 g/L) fed by nipple bottle, or colostrum supplementation (COL; n = 34): a mixture of 1 L bovine colostrum replacer (125 g/L) and 3 L milk replacer (150 g/L) with 3.8 L of the mixture fed by nipple bottle. Serum IgG was measured within 48 h of birth and body weight was taken at d 0, 57, 60, 64, 70, 77, and 84. Starter intake and bovine respiratory disease (BRD) score were measured daily from d 50-70 and fecal consistency was examined daily from d 56-70. Serum ß-hydroxybutyrate (BHB) and lung consolidation were evaluated at d 57, 64, and 70 and intestinal permeability was assessed by recovery of chromium-EDTA, lactulose, and d-mannitol from plasma after oral administration at d 56 and 65. There was no difference in body weight between treatment groups at the start of weaning, but COL were 2.79 kg (95% CI: 0.90-4.68) and 2.76 kg (95% CI: 0.86-4.65) heavier than CON at d 77 and 84, respectively. Additionally, COL tended to gain 100.00 g/d more than CON calves (95% CI: -10.41-207.13) from d 57-84. There were no differences in any of the other variables measured. Supplementation of bovine colostrum replacer during weaning may improve weight gain, but the mechanism of action is not clear.

Genesis of clinical and subclinical endometritis in dairy cows.

In brief: Clinical and subclinical endometritis are different manifestations of reproductive tract inflammatory disease in dairy cows. This review addresses the genesis of clinical and subclinical endometritis considering metabolic stress, innate immune dysfunction, and shifts in the composition of the uterine microbiota in the postpartum period. Abstract: Up to half of dairy cows may develop one or more types of reproductive tract inflammatory disease within 5 weeks after calving. Clinical endometritis (CE) results from uterine bacterial dysbiosis with increased relative abundance of pathogenic bacteria associated with luminal epithelial damage. These bacteria cause endometrial stromal cell lysis, followed by massive polymorphonuclear neutrophil (PMN) migration, and pyogenesis. CE is defined as endometrial inflammation accompanied by purulent discharge. Purulent discharge is not always accompanied by uterine inflammation (being (rarely) vaginitis or (commonly) cervicitis), hence referred to as purulent vaginal discharge (PVD). Subclinical endometritis (SCE) is an asymptomatic uterine disease defined by a threshold of PMN on cytology that is associated with worse reproductive performance; it has not been linked with bacterial dysbiosis. Current evidence suggests that SCE is a result of metabolic and inflammatory dysfunction that impairs innate immune function and the ability of endometrial PMN to undergo apoptosis, necrosis, and ultimately achieve resolution of inflammation. CE and SCE are diagnosed between 3 and 5 weeks postpartum and commonly overlap, but they are considered distinct manifestations of reproductive tract inflammatory disease. This review addresses the genesis of CE and SCE in postpartum dairy cows considering metabolic stress, innate immune dysfunction, and shifts in the composition of the uterine microbiota.

Invited review: Maximizing value and minimizing waste in clinical trial research in dairy cattle: Selecting interventions and outcomes to build an evidence base.

Clinical trials are a valuable study design for evaluating interventions when it is ethical and feasible for investigators to randomly allocate study animals to intervention groups. Researchers may choose to evaluate the comparative efficacy of intervention groups for their effect on outcomes that are relevant to the specific objectives of their trial. However, the results across multiple trials on the same intervention and with the same outcome should be considered when making decisions on whether to use an intervention, because the results of a single trial are subject to sampling error and do not reflect all biological variability. The objective of this review was to provide an overview of important concepts when selecting intervention groups and outcomes within a randomized controlled trial, and when building a body of evidence for intervention efficacy across multiple trials. Empirical evidence is presented to highlight that integrating and interpreting the efficacy of an intervention across trials is hindered by a lack of replication of interventions across trials. Inconsistency in the outcomes and their measurement among trials also limits the ability to build a body of evidence for the efficacy of interventions. The development of core outcome sets for specific topic areas in dairy science, updated as necessary, may improve consistency across trials and aid in the development of a body of evidence for evidence-based decision-making.

Publication and accessibility of results of controlled trials in dairy science.

Research allows for the discovery of new knowledge and is integral to evidence-based decision-making. However, research is only useful if it is available. The aim of this study was to explore publication and accessibility of full-text reports for controlled trials (experimental studies) conducted in dairy cattle. We determined the proportion of controlled trials presented as abstracts at the 2015 Joint Annual Meeting of the American Dairy Science Association and the American Society of Animal Science or the 2015 American Association of Bovine Practitioners Annual Conference that were subsequently published. Factors associated with publication or non-publication in a peer-reviewed journal were evaluated using risk ratios. For trials that were subsequently published, we compared the sample size, numerical results, and inference between the conference abstract and the subsequent publication. Approximately half of the trials (177 out of 380) reported at conferences were subsequently published. Source conference, whether the conference abstract results were described as preliminary, whether there was at least one positive outcome, author affiliation, whether the trial involved deliberate disease induction, and total sample size were not strongly associated with subsequent publication. For trials that were published, the sample size differed between the conference proceedings and full publications for 22%, the numerical results differed in 29%, and the inference differed for 11%. We also evaluated whether trials included in 9 recent systematic reviews were in English and were available without subscription or cost. Of the 390 trials included in recent systematic reviews, approximately 40% were available only through subscription or access fee. These results suggest that publication and accessibility of research results is suboptimal, representing an area of wastage in dairy cattle research. Researchers should ensure that they publish the results of trials comprehensively in searchable publications, even if the results are not novel or do not detect expected differences, and, when possible, make the results available freely.

Reproductive tract size and position score: Estimation of genetic parameters for a novel fertility trait in dairy cows.

The dairy industry is moving toward selecting animals with better fertility to decrease the economic losses linked to reproductive issues. The reproductive tract size and position score (SPS) was recently developed in physiological studies as an indicator of pregnancy rate and the number of services to conception. Cows are scored as SPS 1, 2, or 3 based on the size of their reproductive tract and its position in the pelvis, as determined by transrectal palpation. The objective of this study was to estimate genetic parameters for SPS to assess its potential as a novel fertility trait. Phenotypes were collected at the University of British Columbia's research herd from 2017 to 2020, consisting of 3,247 within- and across-lactation SPS records from 490 Holstein cows. A univariate animal model was used to estimate the variance components for SPS. Both threshold and linear models were fit under a Bayesian approach and the results were compared using the Spearman rank correlation (r) between the estimated breeding values. The 2 models ranked the animals very similarly (r = 0.99), and the linear model was selected for further analysis. Genetic correlations with other currently evaluated traits were estimated using a bivariate animal model. The posterior means (± posterior standard deviation) for heritability and repeatability within- and across-lactation were 0.113 (± 0.013), 0.242 (± 0.012), and 0.134 (± 0.014), respectively. The SPS showed null correlations with production traits and favorable correlations with traditional fertility traits, varying from -0.730 (nonreturn rate) to 0.931 (number of services). Although preliminary, these results are encouraging because SPS seems to be more heritable than and strongly genetically correlated with number of services, nonreturn rate, and first service to conception, indicating potential for effective indirect selection response on these traits from SPS genetic selection. Therefore, further studies with larger data sets to validate these findings are warranted.

Graduate Student Literature Review: What do we know about the effects of clinical and subclinical hypocalcemia on health and performance of dairy cows?

This narrative literature review summarizes findings regarding the associations of clinical and subclinical hypocalcemia with postpartum health, reproduction, and milk production. To better understand the effects of hypocalcemia, we reviewed clinical and subclinical presentations of the condition and the dynamics of blood Ca concentration in the early postpartum period. We summarize and discuss the associations between hypocalcemia and performance of dairy cows. Up to 50% of dairy cows suffer from at least one disease event in the transition period. The important roles of calcium in muscle contraction and immune function make it a key component of metabolism, inflammation, and defense against infection. Although the effect of clinical hypocalcemia (milk fever) on health and performance is clear, the definition of subclinical hypocalcemia (SCH) and its consequences for health and performance are still ambiguous. Differences in study designs, sampling protocols, Ca concentration thresholds, and sample sizes that may be underpowered for health and reproduction outcomes lead to inconsistent conclusions on the effects of SCH. On current evidence, classification of SCH should be based on at least 2 measurements of blood calcium, using cutpoints supported with relevant data, which may vary depending on the outcome of interest. Arbitrary or poorly supported interpretative thresholds for blood Ca concentrations should be abandoned. Transient SCH appears to be associated with greater milk yield, whereas SCH that is present several days after calving is associated with lesser production and greater disease risk. However, when blood calcium is measured days after calving, primary effects of calcium metabolism may be confounded by feed intake, inflammation, or disease, which might either contribute to or be a consequence of hypocalcemia. Additional research is needed to refine sampling schemes to classify SCH, and to better inform the goals and means of prevention of SCH.

The effect of pegbovigrastim on early-lactation disease, production, and reproduction in dairy cows.

The objective of this randomized, double-blind, controlled trial was to evaluate the effect of pegbovigrastim (pegylated recombinant bovine granulocyte colony-stimulating factor) on early-lactation disease, milk yield, and reproduction on commercial dairy farms. A total of 1,607 Holstein cows from 6 farms in Ontario and Québec, Canada, were randomly assigned to receive two 2.7-mL subcutaneous injections of either 15 mg of pegbovigrastim (n = 798; Imrestor, Elanco) or sterile physiological saline (placebo; n = 809). The first injection was administered by investigators 1 wk before expected calving, and the second by farm personnel within 24 h after calving, according to the product label. Producers inspected cows daily and using standardized disease definitions, recorded cases of retained placenta, metritis, displaced abomasum, and clinical mastitis until 63 d in milk. Progesterone concentration was measured in serum at wk 3, 5, 7, and 9 postpartum. Cows were examined for purulent vaginal discharge using the Metricheck (Simcro) device and endometritis using the cytobrush method at wk 5 postpartum. Milk production and reproduction data were obtained from farm management software and the national milk recording database. Disease and culling outcomes were assessed with logistic regression, milk production with linear regression, and time-to-event outcomes with proportional hazards regression. All analyses considered parity and pre-treatment body condition score and their interaction with treatment, and accounted for clustering of cows within farm. In a subset of 246 cows, the effect of treatment on metabolic markers (serum concentrations of glucose, ß-hydroxybutyrate, nonesterified fatty acids, cholesterol, haptoglobin, albumin, and calcium) was assessed in wk 1 and 2 postpartum. Pegbovigrastim had no significant effects on the incidence of retained placenta, metritis, displaced abomasum, clinical or subclinical mastitis, purulent vaginal discharge, or endometritis. Treatment reduced the serum concentration of glucose, slightly reduced the concentration of albumin, and slightly increased concentrations of ß-hydroxybutyrate and nonesterified fatty acids, with no effect on the other markers. There were no differences between treatments in culling risk, time to first insemination, pregnancy at first insemination, or time to pregnancy. Milk yield over the first 3 test days of lactation was 1.0 kg per day lower in the pegbovigrastim group, although a mechanism for that effect could not be explained through analysis of our data.

Controlled trial of the effect of negative dietary cation-anion difference prepartum diets on milk production, reproductive performance, and culling of dairy cows.

Our objective was to assess the effects of feeding negative dietary cation-anion difference (DCAD) prepartum diets on milk production, reproductive performance, and culling. Cows from 4 commercial farms in Ontario, Canada were enrolled in a pen-level controlled trial from November 2017 to April 2019. Close-up pens (1 per farm) with cows 3 wk before calving were randomly assigned to a negative DCAD (TRT; -108 mEq/kg of dry matter; target urine pH 6.0-6.5) or a control diet (CON; +105 mEq/kg of dry matter with a placebo supplement). Each pen was fed TRT or CON for 3 mo (1 period), and then switched to the other treatment for the next period (4 periods per farm). Data from 15 experimental units (8 pen treatments in TRT and 7 in CON), with a total of 1,086 observational units (cows), were included. The effect of treatment on milk yield at the first 3 milk recording tests of lactation was assessed with linear regression models accounting for repeated measures. The risk of pregnancy at first artificial insemination and culling by 30, 60, and 305 d in milk (DIM) were analyzed with logistic regression models, and effects on time to first AI, pregnancy, and culling were assessed with Cox proportional hazards models. All models included treatment, parity, and their interactions, accounting for pen-level randomization and clustering of animals within farm with random effects, giving 10 degrees of freedom for treatment effects. Multiparous cows fed TRT produced more milk at the first (42.0 vs. 38.8 ± 1.2 kg/d) and second (44.2 vs. 41.7 ± 1.3 kg/d) milk tests. However, multiparous cows fed TRT tended to have 0.2 percentage units less milk fat content at these tests. Although multiparous cows fed TRT tended to have greater energy-corrected milk at the first test (least squares means ± standard error: TRT = 46.1 ± 0.9 vs. CON = 43.8 ± 1 kg/d), there were no differences observed in energy-corrected milk at the second or third tests. In primiparous cows, there was no effect of treatment on milk production. Multiparous cows fed TRT had greater pregnancy to first insemination (TRT = 42 ± 3 vs. CON = 32 ± 4%) and tended to have shorter time to pregnancy [hazard ratio (HR) = 1.20; 95% CI: 0.96-1.49]. In primiparous cows fed TRT, time to pregnancy was increased (HR = 0.76; 95% CI: 0.59-0.99). Culling by 30 DIM tended to be less in TRT (3.3 ± 1.1%) than CON (5.5 ± 1.8%). No effect of treatment on culling by 305 DIM was detected in primiparous cows, but in multiparous cows, the TRT diets decreased the odds of culling (21.3 ± 1.9 vs. 31.7 ± 2.8%) and daily risk of culling to 305 DIM (HR = 0.64; 95% CI: 0.46 to 0.89). Under commercial herd conditions, prepartum negative DCAD diets improved milk production and reproductive performance, and reduced culling risk in multiparous cows. In primiparous cows, TRT diets had no effect on milk yield or culling, but increased the time to pregnancy. Our results suggest that negative DCAD diets should be targeted to multiparous cows.

Controlled trial of the effect of negative dietary cation-anion difference on postpartum health of dairy cows.

The objective of this study was to assess the effects of feeding negative dietary cation-anion difference (DCAD) dry cow diets on postpartum health. Cows from 4 commercial dairy farms in Ontario, Canada, were enrolled in a pen-level controlled trial from November 2017 to April 2019. Close-up pens (1 per farm), with cows 3 wk before expected calving, were randomly assigned to a negative DCAD [TRT; -108 mEq/kg of dry matter (DM); target urine pH 6.0-6.5] or a control diet (CON; +105 mEq/kg of DM with a placebo supplement). Each pen was fed TRT or CON for 3 mo (1 period) then switched to the other treatment for the next period, with 4 periods per farm. Urine pH was measured weekly until calving, and body condition score (BCS) was measured at enrollment and at 5 wk postpartum. Data from 15 experimental units [8 TRT and 7 CON, with 1,086 (TRT: n = 681; CON: n = 405) observational units (cows)] that received the assigned diet for >1 wk were included. The incidence of milk fever (MF), retained placenta (RP), metritis, hyperketonemia (blood ß-hydroxybutyrate >1.2 mmol/L, measured weekly in wk 1 and 2), clinical mastitis within 30 DIM (MAST), displaced abomasum (DA) within 30 d in milk (DIM), purulent vaginal discharge (PVD, assessed once at wk 5), and number of disease events (≥1 or ≥2) were analyzed with logistic regression models with treatment, parity, BCS, and their interactions, accounting for pen-level randomization and clustering of animals within farm with random effects, giving 10 degrees of freedom to test treatment effects. Multiparous cows fed TRT had greater blood calcium between 1 and 4 DIM than multiparous cows fed CON, and the prevalence of subclinical hypocalcemia (total Ca ≤2.14 mmol/L) was lesser when fed TRT compared with CON (d 1: 73 ± 6% vs. 93 ± 4%; d 2: 65 ± 7% vs. 90 ± 5%), with no differences between treatments detected in primiparous cows. We detected interactions of treatment and BCS at enrollment for MF in multiparous cows and of treatment and parity for ≥2 disease events. Overconditioned (BCS ≥3.75) multiparous cows had reduced incidence of MF when fed TRT (TRT: 2 ± 1%, vs. CON: 13 ± 8%). We detected no treatment effects on RP, metritis, hyperketonemia, or PVD incidence. Cows fed TRT had lesser incidence of DA (1.7 ± 0.7% vs. 3.6 ± 1.6%) and tended to have lesser incidence of MAST compared with CON (1.8% ± 0.6% vs. 4.4 ± 1.4%). No treatment effect was detected on ≥1 disease events (TRT: 38 ± 7%, vs. CON: 42 ± 8%); however, multiparous cows on TRT were less likely to have ≥2 disease events than cows on CON (14 ± 4% vs. 23 ± 6%). Under commercial herd conditions, feeding prepartum diets with negative DCAD improved several measures of postpartum health.

The effect of prepartum negative dietary cation-anion difference and serum calcium concentration on blood neutrophil function in the transition period of healthy dairy cows.

Our objectives were to assess the effects of a diet with a negative dietary cation-anion difference (DCAD) before calving on phagocytosis (Pc) and oxidative burst (OB) function of circulating neutrophils, and to determine the associations of serum ionized (iCa) and total calcium (tCa) concentrations with Pc and OB in transition dairy cows. We hypothesized that multiparous cows fed a negative DCAD diet prepartum would have greater iCa and tCa, and thus improved Pc and OB. From 3 wk before expected parturition until calving, 38 healthy multiparous cows from 3 farms were assigned to negative DCAD treatment (TRT; -100 mEq/kg of diet dry matter; n = 21) or a control (CON; 95 mEq/kg of dry matter; n = 17) diet. Each farm was on one treatment or the other at a time, but all farms contributed cows to both groups. Urine pH was measured weekly and in TRT was 6.1 ± 0.8 with 80% of 50 samples <7 and 74% ≤ 6.5. Phagocytosis, OB, iCa, and tCa were measured at d -7, 1, and 4 relative to calving. Median fluorescence intensity for Pc (MFIP) and OB (MFIOB), and the shift of percentage of cells active for Pc (PPc) and OB (POB) were measured in isolated, stimulated neutrophils via flow cytometry. Outcomes were assessed with mixed linear regression models accounting for repeated measures. There were no differences between treatments in the 4 neutrophil function outcomes. Although MFIOB varied over time, there were no interactions of treatment with time for any outcome. Serum ionized and tCa did not differ between TRT and CON. The least squares means ± standard deviation for iCa were: d -7, 1.23 ± 0.12 vs. 1.21 ± 0.12; d 1, 1.07 ± 0.12 vs. 1.02 ± 0.12; d 4, 1.16 ± 0.12 vs. 1.17 ± 0.12 mmol/L for TRT and CON, respectively; and for tCa: d -7 2.39 ± 0.25 vs 2.44 ± 0.31; d 1, 2.01 ± 0.25 vs 1.97 ± 0.31; d 4, 2.33 ± 0.25 vs 2.32 ± 0.31 mmol/L, respectively. The proportion of blood samples with tCa <2.15mmol/L at d -7, 1 and 4 was 5, 76, and 13%, respectively, with no differences between TRT and CON. Correlations of iCa or tCa with each of the 4 polymorphonuclear leukocyte (PMN) function outcomes were weak (r < |0.3|). We did not observe the hypothesized differences in aspects of innate immunity in clinically healthy multiparous cows fed a negative DCAD. We underline that cows that experienced clinical disease were excluded from this study, which is important for interpretation of the results.

Invited review: Completeness of reporting of experiments: REFLECTing on a year of animal trials in the Journal of Dairy Science.

Reproducibility is an essential element of the scientific process, and it requires clear and complete reporting of study design, conduct, and analysis. In the human and animal health literature, incomplete reporting is associated with biased effect estimates. Moreover, incomplete reporting precludes knowledge synthesis and undervalues the resources allocated to the primary research. The Reporting Guidelines for Randomized Controlled Trials for Livestock and Food Safety (REFLECT) statement, published in 2010, is a checklist developed by expert consensus to provide guidance on what study elements should be reported in any intervention trial (designed experiment) involving livestock. The Journal of Dairy Science (JDS) has recently endorsed the use of reporting guidelines. To assess the status of reporting of controlled experiments in JDS and to provide a baseline for future comparison, we evaluated the reporting of 18 items from the REFLECT statement checklist in a sample of 137 controlled trials published in JDS in 2017. Two reviewers independently screened titles and abstracts for relevance and then evaluated a sample of 120 papers reporting controlled trials (experimental studies involving at least one intervention and one comparison or control group), using yes or no questions. Although some items, such as treatment details and statistical analysis, were well reported, other areas, including sample size justification, allocation concealment, blinding, study flow, baseline data, and ancillary analyses, were often not reported or were incompletely described. This work highlights the need for authors and reviewers to take advantage of guidelines and checklists for reporting. Adherence to reporting guidelines can help improve the completeness of reporting of research, expedite and better inform the peer-review process, increase clarity for the reader, and allow for knowledge synthesis, such as meta-analysis, all of which serve to increase the value of the work conducted.

Effect of stall design on dairy calf transition to voluntary feeding on an automatic milk feeder after introduction to group housing.

Automatic milk feeders (AMF) for young dairy calves are widely used in the dairy industry. These feeders are thought to have benefits for calf health and welfare and may reduce labor required for feeding; however, little is known about how calves adapt to feeding with AMF. The objective of this study was to observe the effects of feeding stall design on calves learning to use the AMF. The hypothesis was that solid side stalls, compared with steel bar stalls, would result in a longer latency to approach and feed from the AMF without assistance. A total of 147 Holstein calves (80 male and 67 female) were enrolled at 4 d of age, introduced to a group pen, and, at the same time, trained on an AMF. For training, calves were allowed to suck on the trainer's fingers and guided to the teat. Calves were allocated to 1 of 2 stall designs at the pen level, depending on which treatment cohort they were born into, either with steel bar stall walls (n = 46 male, 34 female calves) or with solid side stall walls (n = 34 male, 33 female calves). For 72 h after introductory training on the AMF, data from the feeders were collected and calf behavior was monitored by video. Outcomes measured included latency to first voluntary visit to the feeder and to first feeding, time spent in the feeder, amount of milk consumed over 72 h, number of retraining sessions required (retrained if <2 L was consumed every 12 h), and exploratory behavior, such as sniffing and licking of the feeder. Data were analyzed using mixed effects linear regression models or a Poisson model for the outcome of retraining. For certain outcomes the effects of stall design interacted with difficulty of training (willingness to enter feeder and drink); for the 38% of calves that were scored as moderately difficult to train on a scale of easy, moderate, or difficult, treatment (stall design) differences were detected. These calves took 2× longer to lick or bite toward the nipple, 2× longer to first voluntarily feeding, and consumed less milk over 72 h following training when trained on the steel bar stall design. These results suggest simple features of a stall may influence how quickly calves learn to use an AMF, but that the influence of stall wall design was affected by how easy calves were to train on the feeder upon initial introduction, which may depend in part on certain aspects of calf temperament. For many calves, solid side stalls at an AMF resulted faster in adaption than the steel bar stalls.

The effect of prepartum feeding and lying space on metabolic health and immune function.

The determinants of metabolic and reproductive health disorders in the peripartum period and the degree to which feeding and lying space and management can influence health are only partially understood. The objective of this randomized controlled study was to determine whether providing noncompetitive feeding and lying access in the close-up dry period improves health and immune function. Forty-eight Holstein cows of all parities were randomly assigned to a treatment group of 6 to 10 cows in 1 pen with either 80% cows to stalls and 90 cm of feeding space per cow (understocked) or 120% stocking density and 45 cm of feeding space per cow (overstocked) for 3 wk before expected calving. All cows wore electronic data loggers to monitor daily standing and lying time. Video recordings representing d 7 to 9 after group formation were reviewed, and a competition index (C_Ind) was calculated for each cow by dividing the number of times a cow displaced another as an actor by its total number of actor and reactor displacements. Cows were categorized as high success (C_Ind ≥0.6), moderate success (0.4 ≤ C_Ind <0.6), or low success (C_Ind <0.4). Weekly blood samples measured nonesterified fatty acids, ß-hydroxybutyrate, calcium, glucose, albumin, aspartate aminotransferase, bilirubin, haptoglobin, insulin, and insulin-like growth factor-1 from 3 wk before to 5 wk after calving. Measures of innate immune function (neutrophil phagocytosis and oxidative burst) were assessed at -2, -1, 1, 2, 3, and 5 wk relative to calving. Liver biopsies were collected at wk 1 and 3. Cows in the understocked group spent significantly more time per day lying; the back-transformed least squares means and 95% confidence interval were 14.8 h (13.9-15.6) versus 12.8 h (12.0-13.7). Controlling for parity, there was no difference between treatments in ß-hydroxybutyrate, nonesterified fatty acids, glucose, insulin, insulin-like growth factor 1, aspartate aminotransferase, bilirubin, or haptoglobin concentrations. Throughout the study, cows in the understocked treatment had higher mean calcium and tended to have higher albumin and at 3 wk after calving tended to have lower mean liver triacylglycerol content. Overall, there was no treatment effect on phagocytosis, but cows with a higher C_Ind in the understocked treatment group had greater oxidative burst function. There was no effect of treatment on endometritis. Despite increased competition and lower lying time, the expected harmful effects of crowding and competition on metabolic indicators and innate immune function were mostly not observed. Although this does not refute the importance of access to feeding and lying space, these results indicate that metabolic and reproductive health is more complex than can be explained solely by exposure to what are understood to be best practices for space allowances.

Short communication: The effect of calcium supplementation at calving on neutrophil function.

Low total blood calcium concentration after calving has been demonstrated to be a risk factor for reduced neutrophil function. The objective of this study was to evaluate whether administration of an injectable calcium supplement product soon after calving increased neutrophil oxidative burst or phagocytosis capacity. Cows (n = 27) from 4 farms were blocked by parity and randomly assigned to receive either calcium gluconate (35% wt/vol) in combination with calcium glucoheptonate (10% wt/vol; Theracalcium, Vétoquinol Canada Inc., Lavaltrie, Quebec, Canada) or a placebo within 12 h after calving and again 24 h later. Each dose of 120 mL was injected subcutaneously over 2 sites. Total serum calcium concentration, neutrophil oxidative burst, and neutrophil phagocytosis capacity were measured from coccygeal blood samples before (time 0) and 72 h after first treatment. There was no difference between treatment groups in lactation number, total calcium concentration, oxidative burst, or phagocytosis at time of enrollment. There was no effect of treatment on oxidative burst or phagocytosis by neutrophils. This preliminary study does not support an effect of supplemental calcium to improve neutrophil oxidative burst or phagocytosis capacity of low-parity parturient cows.

Comparison of online, hands-on, and a combined approach for teaching cautery disbudding technique to dairy producers.

The use of pain control for disbudding and dehorning is important from both an animal and industry perspective. Best practices include the use of local anesthetic, commonly given as a cornual nerve block (CNB), and a nonsteroidal anti-inflammatory drug. The proportion is decreasing, but many dairy producers do not use local anesthesia, perhaps in part due to lack of knowledge of the CNB technique. Although this skill is typically learned in person from a veterinarian, alternative methods may be useful. The objective of this trial was to determine if there were differences in the efficacy of online training (n = 23), hands-on training (n = 20), and a combined approach (n = 23) for teaching producers to successfully administer a CNB and disbud a calf. The primary outcome was block efficacy, defined as a lack of established pain behaviors during iron application. Secondary outcomes were background knowledge (assessed by a written quiz), CNB and disbudding technique (evaluated by rubric scoring), time taken, and self-confidence before and after evaluation. Associations between training group and outcome were assessed with logistic regression, ordered logistic regression, and Cox-proportional hazard models, with a random effect for workshop. Block efficacy was not different between training groups, with 91% successful in both combined and online groups, and 75% in the hands-on trained group. Online learners had poorer technical scores than hands-on trainees. The combined group was not different from hands-on. Time to block completion tended to be longer for the online group (62 ± 11 s), whereas time to disbudding completion was not different between hands-on (41 ± 5 s) or combined trainees (41 ± 5 s). The combined group had the highest pre-evaluation confidence score, and remained higher after evaluation than online but was not different than hands-on. Although we saw some statistical differences between groups, absolute differences were small and block efficacy was similar. This suggests online training can be a useful tool for motivated producers who lack access to hands-on training.

Effects of local anesthetic or systemic analgesia on pain associated with cautery disbudding in calves: A systematic review and meta-analysis.

Disbudding is a common management procedure performed on dairy farms and, when done without pain mitigation, is viewed as a key welfare issue. Use of pain control has increased in recent years, but full adoption of anesthesia and analgesia by veterinarians or dairy producers has not been achieved. This may in part be due to the lack of a consistent recommendations of treatment protocols between studies examining pain control methods for disbudding. The objective of this systematic review was to examine the effects of these pain control practices for the most common method of disbudding, cautery, on outcomes associated with disbudding pain in calves. The outcomes were plasma cortisol concentrations, pressure sensitivity of the horn bud area, and validated pain behaviors (ear flick, head shake, head rub, foot stamp, and vocalization). Intervention studies describing cautery disbudding in calves 12 wk of age or younger were eligible, provided they compared local anesthesia, nonsteroidal anti-inflammatory drug (NSAID), or local anesthesia and NSAID to 1 or more of local anesthesia, NSAID, or no pain control. The search strategy used the Agricola, Medline (via OvidSP), and Web of Science databases, as well as the Searchable Proceedings of Animal Conferences (S-PAC), ProQuest Dissertations and Theses Database, and Open Access Theses and Dissertations. Meta-analysis was performed for all outcomes measured at similar time points with more than 2 studies. Local anesthetic was associated with reduced plasma cortisol until 2 h postdisbudding; however, a rise in cortisol was observed in the meta-analysis of studies reporting at 4 h postdisbudding. Heterogeneity was present in several of the analyses for this comparison. The addition of NSAID to local anesthetic showed reduction in plasma cortisol at 4 h, and a reduction in pressure sensitivity and pain behaviors in some analyses between 3 and 6 h postdisbudding. Heterogeneity was present in some meta-analyses, including several using pain behavior outcomes. This may reflect the variation in measurement time periods for behavioral measures between studies, as well as differences among NSAID treatments. Overall, a protective effect of local anesthetic was seen for the acute pain of cautery disbudding, and the delayed rise in cortisol was mitigated by the addition of an NSAID, which also reduced other signs of pain, including pressure sensitivity and pain behaviors. Based on these findings, we recommend use of local anesthetic and an NSAID as best practices for pain mitigation for cautery disbudding of calves 12 wk of age or less. The magnitude and duration of the effect of NSAID treatment was not possible to deduce from the literature because wide variation existed between studies. We recommend consideration of more standardized outcome measurements, especially for pain behaviors. Adherence to reporting guidelines by authors would help ensure more transparent and complete information is available to end users.