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.

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.

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.

Validation of a point-of-care handheld blood total calcium analyzer in postpartum dairy cows.

Our objective was to validate a point-of-care handheld blood total calcium analyzer (Ca meter, CM; TD-5220 Vet Ca2+, TaiDoc, New Taipei, Taiwan) to estimate circulating Ca concentrations in postpartum dairy cows. Whole blood was collected from 251 multiparous cows between 1 and 4 d in milk from 2 commercial dairy herds in Ontario, Canada. Blood total calcium concentration (tCa) was analyzed in whole blood, fresh plasma, and thawed plasma, and compared with tCa results from thawed serum analyzed in a diagnostic laboratory (using a Cobas Calcium Gen 2 kit, Roche Diagnostics, Indianapolis, IN) as the reference test (RT). Lin's concordance correlation coefficient (ßrho;) and Bland-Altman (B-A) plots were assessed to evaluate the agreement between the RT and CM results in each type of sample. Receiver operating characteristic curve analyses were used to describe the accuracy of each test against the categorized RT results (at a cut-point of ≤2.14 mmol/L). Samples where the meter gave a nonquantitative result ("high" or "low"; thawed plasma: 3/247; fresh plasma: 6/100; and whole blood: 20/98) were not included in the ßrho; and B-A analyses. Lin's correlation coefficients demonstrated poor agreement between tests (thawed plasma: ßrho; = 0.16; fresh plasma: ßrho; = 0.21; and whole blood: ßrho; = 0.23). Fresh plasma (using a cut-point of 2.55 mmol/L as measured on the CM) had the greatest diagnostic sensitivity (72%), specificity (86%), and accuracy (77%) for determining subclinical hypocalcemia, but that would still misclassify 23% of samples. In addition to substantial variability, the B-A plots revealed bias with changing concentrations of calcium. Because of low sensitivity on whole blood (58%) or thawed plasma (56%), measurement with the CM is not recommended on these types of samples. This rapid and low-cost meter was not sufficiently accurate to quantify blood Ca concentration, but when used with fresh plasma it might be useful as a screening tool for subclinical hypocalcemia.

Dynamics of pre- and post-insemination progesterone profiles and insemination outcomes determined by an in-line milk analysis system in primiparous and multiparous Canadian Holstein cows.

The objective was to evaluate in-line milk progesterone (mP4) data to determine dynamics of pre- and post-insemination mP4 profiles and their associations with parity and outcomes of artificial insemination (AI) in Holstein cows. Milk progesterone (ng/mL) was quantified at pre-determined time points before and after AI through an automated in-line milk analysis system (Herd Navigator™, DeLaval International, Tumba, Sweden). Only AI (∼d0; n = 605) preceded by an mP4-decline (at least two samples of mP4 ≥5 ng/mL followed by at least one sample <5 ng/mL; d-2) were evaluated. Maximum mP4 attained between d-15 and d-2 (PrePeak), d-2, d5, d10, d14, maximum mP4 attained within 21d post-AI (PostPeak), and rate-of-daily-change between mP4 time points (ng/mL/d) were analyzed. Primiparous and multiparous cows were classified by AI outcomes based on post-AI mP4 profiles into three groups: (1) non-pregnant (OPEN; mP4-decline ≤ 30d post-AI), (2) presumed-pregnant (PREG; no mP4-decline until 55d post-AI), and (3) presumed-pregnancy loss (P-LOSS; mP4-decline between 31 and 55d post-AI). For profile comparisons, smoothed mP4 data were analyzed using mixed linear models. Primiparous cows had greater (P < 0.01) mP4 than multiparous cows at d5 (4.6 ± 0.2 vs. 2.8 ± 0.1), 10 (11.1 ± 0.4 vs. 7.6 ± 0.2), 14 (19.7 ± 0.4 vs. 16.1 ± 0.3) and PostPeak (23.5 ± 0.3 vs. 21.7 ± 0.2). The rate-of-daily-change was greater (P < 0.01) in primiparous than in multiparous cows from d-2 to 5 (+0.2 ± 0.03 vs. -0.1 ± 0.02) and from d5 to 10 (+1.2 ± 0.05 vs. +0.9 ± 0.03), but lesser (P < 0.01) from d14 to PostPeak (+0.9 ± 0.09 vs. +1.3 ± 0.06). In primiparous cows, mP4 in PREG was greater at d10 and PostPeak than OPEN (11.1 ± 0.5 and 24.2 ± 0.5 vs. 9.6 ± 0.4 and 22.3 ± 0.4, respectively, P < 0.04), but lesser at d5 than P-LOSS (4.4 ± 0.3 vs. 5.7 ± 0.4, P = 0.04). In multiparous cows, mP4 at d-2 was lesser in PREG than OPEN and P-LOSS (3.2 ± 0.1 vs. 3.4 ± 0.04 and 3.5 ± 0.1, respectively, P ≤ 0.03), but greater at d10, d14 and PostPeak in PREG than in OPEN (8.2 ± 0.4, 16.8 ± 0.5 and 22.7 ± 0.4 vs. 6.9 ± 0.3, 14.8 ± 0.3 and 19.7 ± 0.2, respectively, P < 0.01). Multiparous PREG cows had greater rate-of-daily-change in mP4 than OPEN cows from d5 to 10 and from d10 to 14 (+1.0 ± 0.06 and +2.2 ± 0.11 vs. +0.8 ± 0.04 and +1.9 ± 0.08, respectively, P < 0.03). Overall post-AI mP4 increased faster and were greater in primiparous than in multiparous cows. Based on in-line mP4 profiles, greater mP4 levels near time of AI (d-2 in multiparous and d5 in primiparous cows) and lesser mP4 beyond d10 were negatively associated with a successful pregnancy.

Relationships among early postpartum luteal activity, parity, and insemination outcomes based on in-line milk progesterone profiles in Canadian Holstein cows.

The objectives of this retrospective study were to use in-line milk progesterone (mP4) data to investigate relationships of (1) commencement of luteal activity (CLA), and (2) luteal phase (LP) length and frequency preceding first postpartum AI, with parity and AI outcomes in Canadian Holstein cows. Starting 21 ± 1 days postpartum (DPP), levels of mP4 were assessed every 2.2 ± 2.0 d through an automated in-line milk analysis system (Herd Navigator™, DeLaval International, Tumba, Sweden) until ∼55 d after first or second AI in 748 Holstein cows from two herds. The CLA was defined as the DPP of the first of at least two consecutive samples with mP4 ≥5 ng/mL, and the period with elevated mP4 (≥5 ng/mL) was defined as the LP. Cows were categorized by CLA [earlier (≤) or later (>) than 28, 35, 42, 49, 56, and 63 DPP], and by the pattern of LP frequency preceding first AI as having or not: (1) one or more normal LP (LP length ≥7 and ≤19 d); (2) one or more abnormal LP (LP length <7 or >19 d, or interluteal period ≥12 d); and (3) two or more LP (either normal or abnormal). Outcomes of first or second AI were determined by the interval between AI and cessation of the ensuing LP as: non-pregnant (mP4-decline ≤30 d), presumed-pregnant (no mP4-decline until 55 d), or presumed-pregnancy loss (mP4-decline between 31 and ≤55 d). The odds of pregnancy per AI (P/AI) at 55 d and pregnancy loss were evaluated using generalized linear mixed models. Primiparous cows had lower odds of having CLA ≤28 DPP [Odds ratio (OR) = 0.58, P = 0.002] and one or more abnormal LP (OR = 0.73, P = 0.04) than multiparous cows. In multiparous cows, CLA ≤28 DPP decreased pregnancy loss (OR = 0.48, P = 0.05) and CLA ≤56 DPP increased P/AI (OR = 4.69, P < 0.01) compared to a later CLA. Primiparous and multiparous cows that had one or more normal LP before first AI had increased P/AI (OR = 3.85 and 3.45, respectively, P < 0.01) and reduced pregnancy loss (OR = 0.26 and 0.27, respectively, P < 0.01) than cows without a normal LP. Primiparous cows that had one or more abnormal LP had decreased P/AI (OR = 0.62, P = 0.04) and increased pregnancy loss (OR = 1.64, P = 0.04) compared to those without an abnormal LP. In summary, AI outcomes were improved in multiparous cows that had early CLA and in cows of both parity groups that had at least one normal LP before first AI. However, primiparous cows that had at least one abnormal LP had reduced AI outcomes. Relationships between early postpartum luteal activity and AI outcomes were inconsistent between primiparous and multiparous cows.