Awareness and perceived barriers to the adoption of best management practices for the transportation of lactating cull dairy cows of dairy producers in Ontario.

The objectives of this study were to understand dairy farmers' awareness and perceived barriers to the implementation of best management practices (BMP) for the transportation of lactating culled cows being moved to auction or abattoir. An invitation to participate was emailed to all dairy farms in Ontario, Canada. Responses to this invitation were used to recruit additional participants through criterion purposive snowball sampling. In total, 28 dairy producers participated in one of 5 semi-structured focus groups. All focus groups were audio recorded, transcribed verbatim, and analyzed with applied thematic analysis. Four themes were identified including 1) transfer of responsibility, 2) interpersonal relationships, 3) juggling priorities, and 4) complexities of long-term planning. While individual participant knowledge varied, participants described misconceptions that other producers held surrounding transport duration and distances, or the number of possible sales points for lactating culled cows. Participants did not agree whether the producer's responsibility ended once the cow left their property or if shared responsibility remained among all stakeholders from the farm to the final destination. Participants discussed the importance of existing trusted relationships with local cattle transporters and veterinarians to ensure information on best practices are shared. Participants also discussed how business pressures (e.g., production demands, space limitations) often challenge their ability to dry off lactating cows before transport; however, participants mentioned the degree of milk production may influence the destination of their animal (e.g., direct to slaughter for animals with high milk production). Some participants described a dynamic balance between business pressures and the potential for declining welfare of the animals under consideration to be culled during lactation. Participants posited that producers who did not prioritize proactive herd management and producers nearing retirement had limited long-term planning for culling individual cows, which might lead to transport of vulnerable or unfit animals. Finally, the lack of access to transportation direct to local slaughter was identified as an important barrier to adherence with the BMP for cull cows. In summary, many participants did not have an accurate understanding of what happens to cull cows after departing the farm and were unsure if they maintained a level of responsibility for an animal after being transported from their farm. Given the priority placed by participants on trusted interpersonal relationships, greater availability of peer-to-peer training of farmers and professional accreditation of animal transporters in addition to structural changes to increase local slaughter capacity may increase implementation of BMP and enhance lactating cull cow welfare.

Attitudes and professional values of veterinarians and veterinary students toward positive welfare states for dairy cattle.

Research that involves agricultural animal welfare has typically aimed to improve welfare by decreasing disease, distress, and pain. Positive welfare does not necessarily occur with the absence of suffering but in combination with opportunities for behaviors or affective states desired by animals. Our objectives were to describe Canadian bovine veterinarians' and veterinary students' attitudes, professional normative values, and perceived ability to promote positive welfare for dairy cows, and to explore participants' provided rationale. With an online cross-sectional survey, Canadian veterinary practitioners (n = 78) and veterinary students (n = 148) were asked, on a 7-point Likert scale, about their attitudes, perceived professional normative values, and perceived ability of veterinarians to promote positive welfare for dairy cows. We used an applied thematic analysis approach within the qualitative description methodology to analyze participants' open-ended text responses. Quantitatively, participants had very favorable attitudes (mean ± SE; 6.3 ± 0.04) and perceived favorable values (5.7 ± 0.06) in the veterinary community toward positive welfare opportunities for dairy cows. Three themes were identified to explain the professional normative values: influences from within the veterinary profession, influences from outside the veterinary profession, and personal views of participants. Participants were confident that veterinarians could suggest positive welfare opportunities for dairy cows (6.1 ± 0.06) but were uncertain that the decision to suggest these opportunities to producers was within a veterinarian's control (4.3 ± 0.11) and were not confident that implementation of positive welfare opportunities was under a veterinarian's control (2.1 ± 0.07). Three themes were identified to explain the barriers to veterinarians promoting positive welfare opportunities for dairy cows: not practical to implement, resistance to change, and concern for the animal. Participants stated that many positive welfare opportunities were impractical or expensive to implement. We conclude that positive attitudes and positive professional values exist in the veterinary community toward positive welfare for dairy cows but that much uncertainty exists regarding a veterinarian's ability to influence change to current practices.

Associations of inflammatory and reproductive tract disorders postpartum with pregnancy and early pregnancy loss in dairy cows.

Our objective was to describe associations of postpartum health with pregnancy and pregnancy loss (P-LOSS) from d 19 to 40 after first postpartum artificial insemination (AI) in lactating Holstein cows. In 2 commercial dairy herds in Ontario, Canada, 468 Holstein cows were enrolled 21 ± 3 d before expected parturition when body condition score (BCS) and lameness were assessed. Serum total Ca, haptoglobin (Hp), and nonesterified fatty acids (NEFA) were measured at 2 and 6 ± 2 d in milk (DIM). Blood ß-hydroxybutyrate (BHB) measurement and metritis detection were done at 4, 8, 11, and 15 ± 2 DIM. Cows were examined for endometritis (ENDO; ≥11.5% polymorphonuclear cells in endometrial cytology) and purulent vaginal discharge (PVD) at 35 ± 3 DIM. Lameness was assessed again at 21 and 49 ± 3 DIM and BCS at 63 ± 3 DIM. First postpartum AI occurred primarily (86%) based on detection of estrus by activity monitors, on average (± standard deviation) at 65 ± 9 DIM, and the remaining cows received timed AI at 86 ± 18 DIM. Serum progesterone (P4) was measured on d 8 and 12 after AI, and pregnancy at first AI (P/AI) was estimated by the expression of ISG15 in peripheral blood leukocytes at d 19 after AI and by pregnancy-associated glycoprotein in serum at d 29, 33, and 40 after AI. Each metabolite (Ca, Hp, NEFA, and BHB) was categorized above or below a cut-point identified with receiver operating characteristic curve analysis associated with P/AI confirmed by ultrasound at d 33 from a larger data set. Data were analyzed using multivariable mixed logistic regression models, accounting for parity, health variables, covariates (season at calving and at AI, milk yield at first Dairy Herd Improvement Association test [categorized into terciles], AI method, and DIM), and herd. The proportions of cows classified pregnant at d 19, 29, 33, and 40 after AI were 64%, 54%, 50%, and 45%, respectively. At d 19 after AI, P/AI was less likely in cows diagnosed with ENDO (52% vs. 69%) or PVD (54% vs. 67%). At d 29, P/AI was less likely in cows with Hp ≥1.54 g/L at 2 DIM (38% vs. 55%) or PVD (35% vs. 56%). Both metritis and ENDO were associated with decreased P/AI at d 40 after AI. Cows diagnosed with metritis had greater risk of P-LOSS from d 19 to 29 (43% vs. 22%) or from d 33 to 40 (37% vs. 7%) than cows without metritis. From d 29 to 33, the risk of P-LOSS was greater in cows with NEFA ≥0.73 mM at 2 DIM (13% vs. 5%) or BCS ≤2.75 at 63 DIM (14% vs. 5%). The concentration of P4 on d 8 after AI was positively associated with P/AI at d 29, 33, and 40, and negatively associated with P-LOSS from d 19 to 29. Postpartum health disorders, particularly reproductive tract disease, can have detrimental effects on early pregnancy establishment and on pregnancy maintenance from d 19 to 40 after AI.

Associations of postpartum health with progesterone after insemination and endocrine signaling during early pregnancy in dairy cows.

Our objective was to investigate associations of postpartum health with serum progesterone (P4) concentrations after first artificial insemination (AI), expression of interferon-stimulated genes (ISG) in blood leukocytes, and serum pregnancy-associated glycoprotein (PAG) during early pregnancy in dairy cows. In this observational study, Holstein cows (n = 475) from 2 commercial herds were enrolled 3 wk before expected parturition. Body condition score (BCS) and lameness were assessed at enrollment, and total Ca, haptoglobin, and nonesterified fatty acids (NEFA) were measured in serum at 2 and 6 ± 2 d in milk (DIM). Blood ß-hydroxybutyrate and metritis were evaluated at 4, 8, 11, and 15 ± 2 DIM, and purulent vaginal discharge (PVD) and endometritis based on endometrial cytology were diagnosed at 35 DIM. Onset of cyclicity was assessed by biweekly P4 measurements. The BCS was assessed at enrollment and at 63 DIM, and lameness at enrollment and at 21 and 49 DIM. First AI was based on estrus detection until ∼75 DIM, or synchronization for timed AI thereafter. Serum P4 was measured at d 8 and 12 after first AI; expression of ISG (ISG15 and RTP4) in blood leukocytes was assessed at d 19; and PAG in serum of pregnant cows was measured at d 29, 33, and 40. Multivariable linear regression models were built including health variables (with metabolites categorized via receiver operating characteristic curve analysis) and covariates (season, milk yield, AI method, DIM), accounting for pregnancy status (for ISG and PAG models), repeated measures (for P4 and PAG models), and herd as random. Results are reported comparing cows affected versus unaffected by each predictor. Serum P4 concentrations were lesser at d 8 after AI in cows that had total Ca ≤2.09 mM (3.6 vs. 4.0 ± 0.2 ng/mL) at 2 DIM or a clinical disease (3.7 vs. 4.0 ± 0.2 mg/mL), and P4 tended to be lesser at d 8 in cows that had PVD (3.6 vs. 4.0 ± 0.2 ng/mL) or any uterine disease (3.7 vs. 4.1 ± 0.2 ng/mL). Among cows pregnant at d 29, relative ISG15 expression (fold change) at d 19 was greater in cows that had lameness (5.10 [4.18-6.18] vs. 3.14 [3.10-3.19]) but tended to be lesser in cows that had endometritis (3.20 [2.89-3.54] vs. 4.29 [4.20-4.35]), and relative expression of RTP4 tended to be lesser in cows that had displaced abomasum (1.02 [0.62-1.67] vs. 1.75 [1.73-1.78]). Serum PAG was lesser at d 29 (3.8 vs. 4.2 ± 0.2 sample-to-positive ratio [S:P]) in cows that had NEFA ≥0.73 mM at 6 DIM, and lesser at d 33 (3.9 vs. 4.5 ± 0.2 S:P) and 40 (3.1 vs. 3.7 ± 0.2 S:P) in cows that had prepartum BCS ≥3.75. These findings suggest long-term effects of metabolic or reproductive tract disorders on luteal function after first AI, pregnancy recognition signaling, and placental function during early pregnancy. These associations may partially explain some of the pathways linking postpartum health and fertility in dairy cows.

Increased parity is negatively associated with survival and reproduction in different production systems.

We conducted a retrospective meta-analysis based on individual cow data to assess the associations of parity, level of production, and pasture-based or intensively fed systems with fertility. Our goal was to provide understandings of the role of parity in risks for removal and reproductive failure. Multilevel models were used to evaluate the fixed effects of parity, milk, milk solids, milk fat and protein percentage and yield, and production system [intensively fed (n = 28,675) or predominantly pasture fed (n = 4,108)] on reproductive outcomes. The outcomes were the hazard of not being bred (HNBRED), hazard of pregnancy (HPREG), pregnancy to first breeding (PREG1), and odds of becoming pregnant in a lactation (OPAL). The 32,783 cows were in 13 studies conducted in Australia (14.6% of cows), Canada (2.4% of cows), and the United States (83.0% of cows). There were 38.5% of cows in the sample in parity 1, 27.3% in parity 2, 16.7% in parity 3, 9.0% in parity 4, and 8.6% in parity ≥5. Compared with cows of parity 1, parity ≥5 cows had a greater HNBRED [hazard ratio (HR) = 2.45], lesser HPREG (HR = 0.73), and reduced OPAL (odds ratio = 0.36). However, the parity ≥5 cows had similar PREG1 to other parities except for parity 1. This suggests the possibility of a higher proportion of subfertile parity ≥5 cows than for other parities. Associations between parity and reproductive measures were influenced by the different milk production measures, indicating that milk yield and milk component percentages and yields modified the odds or hazards of successful reproduction. All milk production measures had quadratic associations with OPAL, indicating that either low or high production or concentration of solids within a cohort reduced OPAL. This reduced OPAL reflected a greater HNBRED for lower milk yield and milk protein and fat yielding cows. Both milk yield and milk protein percentage had quadratic associations with HPREG. When centered milk yield was categorized into quartiles, small differences in HPREG existed. A more marked association of milk protein percentage occurred with HPREG, with optimal HPREG at approximately 0.5% above group mean milk protein percentage. Milk fat percentage (HR = 0.901), fat yield (kg/d; HR = 0.78), protein yield (kg/d; HR = 0.71), and milk solids yield (kg/d; HR = 0.84) were all linearly associated with reduced HPREG. Difference in production systems did not have substantive effects on PREG1 but did for HNBRED, HPREG, and OPAL. Estimates of associations of parity with reproductive outcomes HNBRED, HPREG, and OPAL were influenced by milk and milk solids yield; older cows had markedly lower reproductive outcomes. Interestingly, for PREG1, there were few differences among parities and differences were less influenced by milk yield and constituent measures. The marked associations of parity with removal for all reasons, deaths and culling, and reductions in HNBRED, HPREG, and OPAL indicate a need to focus on the physiological changes with parity to produce better strategies to support optimal longevity of cows.

Postpartum health is associated with detection of estrus by activity monitors and reproductive performance in dairy cows.

The objective of this prospective observational study was to investigate associations of postpartum health with estrus detection (ED) by activity monitors and pregnancy outcomes in dairy cows. A total of 1,743 Holstein cows from 2 commercial dairy herds in Ontario, Canada were enrolled 3 wk before expected parturition and examined for health variables until 9 wk postpartum. Body condition score (BCS) and lameness were measured at 3 wk prepartum, and serum concentrations of total Ca, haptoglobin (Hp), and nonesterified fatty acids were measured at 2 and 6 ± 2 d in milk (DIM), and blood ß-hydroxybutyrate (BHB) and metritis were assessed at 4, 8, 11, and 15 ± 2 DIM. Cows were examined for purulent vaginal discharge (PVD) and endometritis (ENDO) by endometrial cytology at wk 5, for lameness at wk 3 and 7, for BCS at wk 9 postpartum, and for time to onset of cyclicity by biweekly serum progesterone (P4) measurements. Additional disease data were obtained from farm records. Reproductive management for first AI was primarily based on ED by activity monitors until at least 75 DIM, and cows not detected in estrus were synchronized. Data were analyzed in multivariable logistic or Cox proportional hazards regression models including blood markers, health variables, potential covariates, and herd as a random effect. Estrus was detected in 77% of primiparous and 66% of multiparous cows between 50 or 55 DIM and 75 DIM. In 1,246 cows, the model-predicted probability of ED (percentage point difference) was lower in cows that had retained placenta (-14%), ENDO (-7%), PVD (-8%), delayed cyclicity (no P4 > 1 ng/mL by wk 9; -12%), or ≥0.5-point BCS loss (-14%) compared with cows without each of these risk factors, and it was negatively associated with blood BHB at 15 DIM. Considering only variables measured on farm (not requiring laboratory analysis), the probability of ED was lower (56 vs. 81%) in cows with >1 risk factor compared with cows without risk factors. The predicted probability of pregnancy at first artificial insemination (percentage point difference) was lower in cows that had ENDO (-7%) or PVD (-7%), and negatively associated with serum Hp at 6 ± 2 DIM. In cows detected in estrus by 75 DIM (n = 888), risk factors for reduced pregnancy rate by 250 DIM (adjusted hazard ratio (AHR); 95% confidence intervals) included difficult calving (AHR: 0.67; 0.45 to 1.00), metritis (AHR: 0.79; 0.61 to 1.01), PVD (AHR: 0.79; 0.65 to 0.97), or lameness (AHR: 0.79; 0.62 to 1.01), and it was negatively associated with serum Hp at 6 ± 2 DIM. Monitoring postpartum health may be used to identify cows that are more or less likely to be detected in estrus by activity monitors and to become pregnant in a timely manner. This would support a selective reproductive management program with targeted interventions.

Effects of source of supplementary trace minerals in pre- and postpartum diets on reproductive biology and performance in dairy cows.

Our objectives were to evaluate the effects of complete replacement of inorganic salts of trace minerals (STM) with organic trace minerals (OTM) in both pre- and postpartum diets on ovarian dynamics, estrous behavior measured by sensors, preimplantation conceptus development, and reproductive performance in dairy cows. Pregnant cows and heifers (n = 273) were blocked by parity and body condition score and randomly assigned to either STM or OTM diets at 45 ± 3 d before their expected calving. Pre- and postpartum diets were formulated to meet 100% of recommended levels of each trace mineral in both treatments, taking into consideration both basal and supplemental levels. The final target concentrations of Co, Cu, Mn, Se, and Zn were, respectively, 0.25, 13.7, 40.0, 0.3, and 40.0 mg/kg in the prepartum diet, and 0.25, 15.7, 40.0, 0.3, and 63.0 mg/kg in the postpartum diet. The STM group was supplemented with Co, Cu, Mn, and Zn sulfates and sodium selenite, while the OTM group was supplemented with Co, Cu, Mn, and Zn proteinates and selenized yeast. Treatments continued until 156 d in milk (DIM) and were assigned to individual cows using automatic feeding gates. Starting at 21 DIM, ultrasonography examinations of the ovaries were performed weekly to determine the presence of a corpus luteum and postpartum resumption of ovarian cyclicity. Cows were presynchronized with 2 injections of PGF2α at 42 and 56 DIM. Estrous behavior was monitored using electronic activity tags that indirectly measured walking activity. Cows detected in estrus after the second PGF2α were inseminated, and those not detected in estrus by 67 DIM were enrolled in a synchronization program. Cows that returned to estrus after artificial insemination (AI) were reinseminated. Pregnancy diagnosis was performed 33 d after AI, and nonpregnant cows were resynchronized. Transcript expression of interferon-stimulated genes in peripheral blood leukocytes was performed in a subgroup of cows (STM, n = 67; OTM, n = 73) on d 19 after AI. A different subgroup of cows (28 STM, 29 OTM) received uterine flushing 15 d after AI for recovery of conceptuses and uterine fluid for analyses of transcriptomics and metabolomics, respectively. In addition, dominant follicle diameter, luteal size and blood flow, and concentration of progesterone in plasma were measured on d 0, 7, and 15 relative to AI. After flushing, PGF2α was given and the dominant follicle was aspirated 2 d later to measure the concentration of trace minerals by mass spectrometry. Estrous behavior, size of the dominant follicle and corpus luteum, concentration of progesterone, time to pregnancy, and proportion of cows pregnant by 100 d of the breeding period did not differ between treatments. A greater proportion of cows supplemented with OTM had a corpus luteum detected before presynchronization (64.3 vs. 75.2%), and primiparous cows supplemented with OTM tended to resume cyclicity earlier than their STM counterparts. Cows supplemented with OTM had a greater concentration of Cu in follicular fluid than cows supplemented with STM (0.89 vs. 0.77 µg/mL, respectively). In pregnant multiparous cows, expression of receptor transporter protein 4 in peripheral blood leukocytes was 42% greater in the OTM group. Conceptuses of the 2 treatments had 589 differentially expressed transcripts, with many indicating advanced conceptus elongation and greater transcript expression of selenoproteins in the OTM group. In pregnant cows, 24 metabolites were more abundant in the uterine fluid of OTM, including spermidine, sucrose, and cholesterol. In conclusion, replacing STM with OTM caused modest improvements to resumption of ovarian cyclicity and important changes in preimplantation conceptus development, but it did not alter conception risk and pregnancy rate.

Associations of parity with health disorders and blood metabolite concentrations in Holstein cows in different production systems.

Data were obtained from studies in Australia, Canada, and the United States using individual cow data from 28,230 Holstein cows to evaluate associations between parity and disease. Our goal was to develop understanding of disease risks for cows of differing parity. We hypothesized that there would be increased risks of disease and changes in metabolite concentrations with increased parity. Parity ≥5 represented 2,533 cows or 9.0%, parity 4 was 9.8% (2,778), parity 3 as 19.0% (5,355), parity 2 as 28.1% (7,925), and parity 1 was 34.1% (9,639) of the sample. Of these cows, 15.5% were in Australia, 14.7% in Canada, and 69.8% in the United States. Lactational incidence (LI) risk of clinical hypocalcemia increased with parity from 0.1% for parity 1 to 13% for parity ≥5 cows. The marked increase suggests profound differences in metabolism with increased parity. The LI of clinical mastitis was 17.4%. The odds of mastitis increased with parity to 2.5 times greater in parity ≥5 than in parity 1. The LI of lameness increased with parity; specifically, the odds of lameness was 5.6 times greater for parity ≥5 than parity 1. Dystocia incidence was 8.7% and greatest for parity 1 cows. The LI of retained placenta was 7.4% and increased with parity, with the odds for parity ≥5 2.3 times greater than for parity 1. The LI of metritis was 10% and of endometritis 14%, with the greatest odds in parity 1. The LI of clinical ketosis was 3.3% with a marked increase in odds with parity. The prevalence of subclinical ketosis was 26.8% with only cows in parity 1 having lower odds than other parities. Parity ≥5 cows had greater odds (odds ratio = 1.7) of respiratory disease than parity 1 cows, which were lesser than other parities. Metabolite concentrations were evaluated in 5,154 Holstein cows in the precalving, calving, and immediate postcalving data sets. Metabolic measures near peak lactation provided 1,906 observations. Concentrations of ß-hydroxybutyrate (BHB) and nonesterified fatty acids increased with parity on d 1 to 3 of lactation and at peak lactation. On d 1 to 3 after calving differences in glucose, nonesterified fatty acids, and BHB indicated a greater reliance on mobilized lipid to export energy to peripheral tissues as BHB for greater parity cows. Differences in concentrations among parity groups were marked at times, for example >0.20 mM in Ca for parity 1 and 2 to parity ≥5 and >0.33 mM for all older parities compared with parity 1 for P on the day of calving. The marked increase suggests profound differences in metabolism with increased parity are probably influenced, in part, by increased production. We found marked differences in concentrations of metabolites with parity that are consistent with reduced reproduction, health, and body condition for higher parity cows. These unfavorable differences in metabolism in Ca, P, glucose, and cholesterol concentrations for higher parity cows also complement the often-substantial differences in disease risk with parity and suggest a need to carefully consider the parity structure in study design. Managers and advisors will need to consider methods to reduce risk of health disorders tailored to cows of different ages.

Characterizing ruminal acidosis risk: A multiherd, multicountry study.

A multicenter observational study was conducted on early lactation Holstein cows (n = 261) from 32 herds from 3 regions (Australia, AU; California, CA; and Canada, CAN) to characterize their risk of acidosis into 3 groups (high, medium, or low) using a discriminant analysis model previously developed. Diets ranged from pasture supplemented with concentrates to total mixed ration (nonfiber carbohydrates = 17 to 47 and neutral detergent fiber = 27 to 58% of dry matter). Rumen fluid samples were collected <3 h after feeding and analyzed for pH, and ammonia, d- and l-lactate, and volatile fatty acid (VFA) concentrations. Eigenvectors were produced using cluster and discriminant analysis from a combination of rumen pH, and ammonia, d-lactate, and individual VFA concentrations and were used to calculate the probability of the risk of ruminal acidosis based on proximity to the centroid of 3 clusters. Bacterial 16S ribosomal DNA sequence data were analyzed to characterize bacteria. Individual cow milk volume, fat, protein, and somatic cell count values were obtained from the closest herd test to the rumen sampling date (median = 1 d before rumen sampling). Mixed model analyses were performed on the markers of rumen fermentation, production characteristics, and the probability of acidosis. A total of 26.1% of the cows were classified as high risk for acidosis, 26.8% as medium risk, and 47.1% as low risk. Acidosis risk differed among regions with AU (37.2%) and CA (39.2%) having similar prevalence of high-risk cows and CAN only 5.2%. The high-risk group had rumen phyla, fermentation, and production characteristics consistent with a model of acidosis that reflected a rapid rate of carbohydrate fermentation. Namely, acetate to propionate ratio (1.98 ± 0.11), concentrations of valerate (2.93 ± 0.14 mM), milk fat to protein ratio (1.11 ± 0.047), and a positive association with abundance of phylum Firmicutes. The medium-risk group contains cows that may be inappetant or that had not eaten recently or were in recovery from acidosis. The low-risk group may represent cattle that are well fed with a stable rumen and a slower rumen fermentation of carbohydrates. The high risk for acidosis group had lower diversity of bacteria than the other groups, whereas CAN had a greater diversity than AU and CA. Rumen fermentation profile, abundance of ruminal bacterial phyla, and production characteristics of early lactation dairy cattle from 3 regions were successfully categorized in 3 different acidosis risk states, with characteristics differing between acidosis risk groups. The prevalence of acidosis risk also differed between regions.

Bayesian estimation of sensitivity and specificity of a milk pregnancy-associated glycoprotein ELISA test for pregnancy diagnosis between 23 and 27 days after insemination in Holstein dairy cows.

Pregnancy diagnosis using pregnancy-associated glycoprotein (PAG) ELISA technology in blood or milk samples is validated from 28 d after insemination in dairy cows. The objective of this study was to estimate the sensitivity (Se) and specificity (Sp) of a commercial milk PAG-based ELISA in Holstein dairy cows between 23 and 27 d after insemination. Milk samples (n = 268) from 257 Holstein dairy cows 23 to 27 d after AI were submitted for PAG ELISA testing. Pregnancy status was confirmed by either a second milk PAG ELISA test conducted between 28 and 50 d after insemination (n = 200) or transrectal ultrasonography performed between 28 and 59 d after insemination (n = 68). A Bayesian latent class model was used to compare the paired results from the test at 23 to 27 d after AI test to the reference test. The latent class model typically used for comparing 2 or more imperfect tests was extended to include the possibility of pregnancy loss between the 23 to 27 d test and the reference test. Informative priors for the probability of pregnancy loss, and for the Se and Sp of the PAG and ultrasonography reference tests were obtained from the scientific literature. Estimated median Se and Sp of the PAG ELISA test conducted between 23 and 27 d after AI were 0.98 (95% credible interval 0.93 to 1.0) and 0.98 (0.89 to 1.0), respectively, when using a standardized corrected optical density threshold of 0.15. Although the accuracy of the test under investigation was excellent, more data will be needed to confirm the optimal diagnostic cut point for PAG in milk for early pregnancy diagnosis in this time window. The optimal timing of pregnancy diagnosis will depend on herd-specific logistics and the action to be taken to re-inseminate nonpregnant cows.

Reducing milking frequency from twice to once daily as an adjunct treatment for ketosis in lactating dairy cows-A randomized controlled trial.

This randomized controlled trial investigated the effects of temporarily reducing milking frequency (MF) on the resolution of ketosis and milk production in dairy cows in early lactation. To detect ketosis [blood ß-hydroxybutyrate (BHB) ≥1.2 mmol/L], Holstein cows were screened daily from 3 to 16 d in milk using a cow-side meter. Cows diagnosed with ketosis (n = 104) were randomly assigned to twice-daily milking (TDM) or reduced to once-daily milking (ODM) for 2 wk, then returned to twice-daily milking. Both treatment groups received a 5-d treatment of an oral propylene glycol drench (PG; 300 g) beginning on the afternoon of the diagnosis; cows received additional 5-d PG treatments if they had a ketotic test result (blood BHB ≥1.2 mmol/L) at least 4 d after finishing the first PG treatment. Blood BHB tests were conducted for the first 3 d after ketosis diagnosis, and then once every 3 d for 21 d of trial (DOT). Milk and milk component data were collected weekly for 15 wk following trial enrollment. The ODM group showed rapidly and markedly decreased blood BHB concentrations (primiparous cows: 1 DOT, 0.92 ODM vs. 1.22 TDM, 15 DOT, 0.55 vs. 0.81 mmol/L; multiparous cows: 1 DOT, 1.01 vs. 1.40, 15 DOT, 0.78 vs. 1.65 mmol/L). In addition, a logistic regression model indicated that ODM cows were less likely to have blood BHB concentrations ≥1.2 mmol/L [primiparous cows: 3 DOT: ODM 1% (95% confidence interval: 0-10%) vs. TDM 43% (30-58%), 15 DOT ODM 0% (0-0.2%) vs. TDM 22% (13-36%); multiparous cows: 3 DOT: ODM 33% (24-44%) vs. TDM 59% (48-69%), 15 DOT ODM 20.9% (13-31%) vs. TDM 64% (53-74%)]. The proportion of ODM cows that required additional treatments of PG were substantially lower than the TDM group (ODM: 39%; TDM: 64%) than the TDM cows during the initial 21-d period. However, during the 2-wk treatment period, cows in the ODM group produced 26% less milk and 25% less energy-corrected milk than the TDM cows. During wk 3 to 15, when all cows were milked twice daily, ODM cows produced less milk (-14%) and energy-corrected milk (-12%) compared with the TDM group. Milk protein percentage was greater, and milk fat percentage and linear score tended to be greater in the ODM group over 15 wk. In conclusion, a 2-wk reduction of MF in ketotic cows from twice to once daily with treatment with PG resolved ketosis and decreased blood BHB concentrations more effectively than treating TDM cows with PG alone. However, the 2-wk MF reduction had immediate and long-term (up to 13 wk after cessation of MF reduction) negative effects on milk production.

An investigation of blood, milk, and urine test patterns for the diagnosis of ketosis in dairy cows in early lactation.

Ketosis in dairy cattle is primarily diagnosed based on the concentrations of ketone bodies in the blood, milk, or urine. Cow-side tests using these fluids are available for rapid detection of elevated concentrations of ketone bodies. Although these tests have been extensively validated, the performance of different tests has not been compared over time. Our objectives were to investigate the relationship between point-of-care diagnostic tests measuring the concentrations of ß-hydroxybutyrate (BHB) in blood (BT; Precision Xtra, Abbott Laboratories), BHB in milk (MT; Keto-Test, Elanco), and acetoacetate (AcAc) in urine (UT; Ketostix, Bayer Corporation) through cases of ketosis. Holstein cows (n = 148) were screened daily for hyperketonemia (HYK; blood BHB ≥1.2 mmol/L) from 3 to 16 d in milk (DIM); moreover, milk and urine samples were collected concomitantly and tested for ketones (ketosis thresholds: 100 µmol/L milk BHB and 5 mg/dL urine AcAc). Of the animals screened (n = 148), 74% were diagnosed with HYK. When diagnosed with HYK, cows were treated with propylene glycol orally once daily for 5 d. After the day of diagnosis (d 0), hyperketonemic cows were retested with BT, MT, and UT for 3 d (d 1, 2, and 3). We assessed the diagnostic test performance and time to ketosis (survival analyses and Cox proportional hazards models) of MT and UT compared with BT. Considering all paired samples (before and after diagnosis of HYK), MT had 61% sensitivity and 91% specificity, whereas the UT had 77% sensitivity and 94% specificity compared with BT. The specificity of MT and UT increased from d 0 to d 1, decreased on d 2, and increased on d 3. The median time to diagnosis of ketosis in blood was 5 DIM (95% CI 5 to 7 DIM); moreover, MT and UT had 2 d greater median time to diagnosis of ketosis compared with the BT [7 DIM (6 to 11 d); and 7 DIM (6 to 13 d), respectively]. We concluded that the UT is a more sensitive predictor of blood BHB concentration than the MT. The UT and MT tests diagnosed ketotic cows approximately 2 d later than the BT. The possible consequences of delay in detection of ketosis in milk and urine should be investigated.

Effects of replacing inorganic salts of trace minerals with organic trace minerals in pre- and postpartum diets on feeding behavior, rumen fermentation, and performance of dairy cows.

Our objectives were to evaluate the effects of complete replacement of supplementary inorganic salts of trace minerals (STM) by organic trace minerals (OTM) in both pre- and postpartum diets on feeding behavior, ruminal fermentation, rumination activity, energy metabolism, and lactation performance in dairy cows. Pregnant cows and heifers (n = 273) were blocked by parity and body condition score and randomly assigned to either STM or OTM diets at 45 ± 3 d before their expected calving date. Both groups received the same diet, except for the source of trace minerals (TM). The STM group was supplemented with Co, Cu, Mn, and Zn sulfates and Na selenite, whereas the OTM group was supplemented with Co, Cu, Mn, and Zn proteinates and selenized yeast. Treatments continued until 156 days in milk and pre- and postpartum diets were formulated to meet 100% of recommended levels of each TM in both treatments, taking into consideration both basal and supplemental levels. Automatic feed bins were used to assign treatments to individual cows and to measure feed intake and feeding behavior. Rumination activity was monitored by sensors attached to a collar from wk -3 to 3 relative to calving. Blood metabolites were evaluated on d -21, -10, -3, 0, 3, 7, 10, 14, 23, and 65 relative to calving. Ruminal fluid samples were collected using an ororuminal sampling device on d -21, 23, and 65 relative to calving, for measurement of ruminal pH and concentration of volatile fatty acids. Cows were milked twice a day and milk components were measured monthly. Cows supplemented with OTM tended to have longer daily feeding time (188 vs. 197 min/d), and greater dry matter intake (DMI; 12.9 vs. 13.3 kg), and had a more positive energy balance (3.6 vs. 4.2 Mcal/d) and shorter rumination time per kg of dry matter (DM; 40.1 vs. 37.5 min/kg of DM) than cows supplemented with STM during the prepartum period. In the postpartum period, OTM increased DMI in multiparous cows (24.1 vs. 24.7 kg/d) but not in primiparous cows (19.1 vs. 18.7 kg/d). The difference in DMI of multiparous cows was more evident in the first 5 wk of lactation, when it averaged 1 kg/d. Milk yield was not affected by treatment in multiparous cows (44.1 vs. 44.2 kg/d); however, primiparous cows supplemented with OTM had lesser yields than primiparous cows supplemented with STM (31.9 vs. 29.8 kg/d). Cows supplemented with OTM had a greater percentage of protein in milk (3.11 vs. 3.17%), reduced concentration of nonesterified fatty acids in serum (0.45 vs. 0.40 mmol/L), and rumination activity (30.1 vs. 27.8 min/kg of DM) than cows supplemented with STM. At the end of the transition period, cows supplemented with OTM had reduced molar proportion of acetate, reduced pH, and tended to have a greater concentration of total volatile fatty acids in ruminal fluid. In conclusion, complete replacement of STM by OTM caused modest changes in rumen fermentation, feeding behavior, energy metabolism, and performance of dairy cows, improving postpartum DMI in multiparous cows and reducing circulating levels of nonesterified fatty acids. The pre-absorptive effects of TM source and the parity specific responses on performance warrant further research.

Neutrophil function and antibody production during the transition period: Effect of form of supplementary trace minerals and associations with postpartum clinical disease and blood metabolites.

Our objectives were to evaluate the effect of the form of supplementary trace minerals-inorganic salts (STM: Co, Cu, Mn, and Zn sulfates and Na selenite) or organic (OTM: Co, Cu, Mn, Zn proteinates, and selenized yeast)-fed at 100% of recommended levels in both pre- and postpartum diets on in vitro phagocytic activity of neutrophils, and in vivo IgG responses to an ovalbumin challenge during the transition period. In addition, we investigated the associations of these immunological responses with incidence of postpartum clinical diseases and the dynamic changes of metabolic markers during the transition period. Pregnant heifers and cows (n = 273) were enrolled at 45 ± 3 d before expected calving, blocked by parity and body condition score, and allocated randomly to STM or OTM supplementation. Cows in both treatments were fed the same diet, except for the form of supplementary trace minerals. Automatic feeding gates were used to assign treatments to individual cows. Blood was collected on d -7 ± 3 and 7 ± 3 relative to calving in a subgroup of cows (n = 131 and 133, respectively) to measure phagocytic activity of neutrophils in vitro using flow cytometry. Subcutaneous immunization with 0.5 mg of chicken egg ovalbumin was performed in a subgroup of cows (n = 181) on d -45, -21, and 3 relative to calving. Concentration of anti-ovalbumin IgG in serum was measured by ELISA on d -45, -21, 3, 7, and 21. Trace mineral concentrations in blood were measured by inductively coupled plasma mass spectrometry on d -45, -21, -7, 0, 7, and 21 relative to calving. Selected metabolites were measured on d -21, -10, -3, 0, 3, 7, 10, 14, and 21 relative to calving. Treatment did not affect the percentage of neutrophils performing phagocytosis on d -7 or 7 but the median fluorescence intensity of phagocytosis on d 7 was greater for OTM than STM. We found no differences between treatments in the level of anti-ovalbumin IgG in serum on any of the sampling days. Changes in neutrophil function from prepartum to postpartum were associated with incidence of postpartum clinical disease, postpartum feed intake and milk production, concentrations of Ca, K, Se, Mn, Co, and total protein in serum. Immunoglobulin G responses to ovalbumin injections were not associated with incidence of postpartum clinical disease but were associated with body weight, feed intake, energy balance, and concentrations of nonesterified fatty acids, aspartate aminotransferase, gamma-glutamyl transpeptidase, albumin, Na, P, and Cu in serum. In conclusion, replacement of STM by OTM improved one measure of phagocytic capacity of neutrophils in vitro, which was also greater in cows that did not develop postpartum clinical disease. The associations of innate and acquired immune responses with feed intake, energy balance, and circulating concentrations of key macro and micronutrients reinforce the importance of nutritional management for the health of dairy cows during the transition period.

Associations of calf management practices with antimicrobial use in Canadian dairy calves.

The objective of this cross-sectional study was to investigate the relationship between management practices and antimicrobial use in heifer calves on Canadian dairy farms. Questionnaires on calf management practices, herd characteristics, and calf treatment records were administered on 147 dairy farms in 5 provinces during annual farm visits in a multiyear, nationwide research project (Canadian Dairy Network for Antimicrobial Stewardship and Resistance: CaDNetASR). Questions focused on the calf caregiver, calving pen, colostrum management, milk feeding, grouping, bedding management, and age when male calves were sold. Antimicrobial treatment records were collected on each farm from either an electronic herd management system or paper-based records. Newborn heifers born in the last 12 mo were identified retrospectively and followed to 60 d of age, with antimicrobial treatments and dates of sale or death extracted for further analysis. A multivariable linear regression model was developed with the natural log of the number of antimicrobial treatments per calf-year as the dependent variable, and categorized calf management practices and farm characteristics as the independent variables. A complete data set of records on 7,817 calves was retrieved from 74 farms based on completeness of calf records. A total of 2,310 calves were treated at least once with an antimicrobial, and 7,307 individual antimicrobial treatments were recorded. Among the reasons for antimicrobial use, respiratory disease (54%) was most common, followed by diarrhea (20%), presence of a fever (3%), and umbilical disease (2%). Florfenicol (33% of recorded treatments), penicillin (23%), and trimethoprim-sulfamethoxazole (18%) were commonly used, whereas fluoroquinolones (4%), and ceftiofur (1%) were used less commonly. Farms (31%) commonly had 0-1.0 antimicrobial treatments/calf-year (median: 2.2 treatments/calf-year; interquartile range: 0.64-6.43 treatments/calf-year). Defined daily dose (DDD) per calf-year was calculated based on the Canadian bovine standards. Among the 74 farms, florfenicol (1.35 DDD/calf-year) and macrolides (0.73 DDD/calf-year) were used most, whereas ceftiofur (0.008 DDD/calf-year) was the lowest. The final multivariable linear regression model indicated that farms that fed transition milk had fewer than half the number of antimicrobial treatments per calf-year than those who did not feed transition milk. The number of antimicrobial treatments per calf-year in preweaning calves was low on many farms, and there was low use of highly important drugs for human medicine. The effect of feeding transition milk should be investigated regarding potential effects on antimicrobial use and disease prevention.

Associations between feeding behaviors collected from an automated milk feeder and disease in group-housed dairy calves in Ontario: A cross-sectional study.

The adoption of automated milk feeders and group housing of preweaning dairy calves has become more common in Canada; however, disease detection in group-housed calves remains a challenge. The aim of this cross-sectional study was to assess whether feeding behavior data collected from a single point in time could be used to aid in the detection of neonatal calf diarrhea (NCD), bovine respiratory disease (BRD), and general disease, in preweaning group-housed calves being fed via an automated milk feeder. The data used was collected in an earlier study. A total of 8 dairy farms recruited from an online survey of calf-management practices were enrolled into the study. There was a total of 523 observations with 130 events of NCD, 115 events of BRD, and 210 events of general disease. Each farm was visited once in each of the fall, winter, spring, and summer, when the calves' health was scored, and the data were collected from the automated milk feeders. Mixed linear regression models were used to identify associations between feeding behavior data (milk consumption, time spent at the feeder, drinking speed, and the number of rewarded and unrewarded visits) and the presence of NCD, BRD, or general disease (having one or more of NCD, BRD, or umbilical infection), on the day of health scoring. Generalized linear mixed models were used to analyze the percentage of milk the calf consumed from their daily milk allotment. Calves with BRD consumed 63% less of their daily allotment of milk, had 2 fewer unrewarded visits to the automated milk feeder, and drank milk 152 mL/min slower compared with calves without BRD. Calves with NCD consumed 57% less of their daily milk allotment, consumed 758 mL less per day, and drank 92 mL/min slower than calves compared with calves without NCD. Calves with general disease drank 50% less of their daily milk allowance, consumed 496 mL less per day, drank 80 mL/min slower, and had 2 fewer unrewarded visits to the automated milk feeder, when compared with calves without disease. No significant associations were found between the presence of NCD, BRD, or general disease and time spent at the feeder or number of rewarded visits. Sensitivity and specificity values for disease identification were low when evaluating the feeding behaviors individually, so parallel testing was completed. To do so, if any significant feeding behavior was below the optimal cut point for disease detection as determined using a ROC curve, the calf was considered positive for disease and the sensitivity and specificity were recalculated. Parallel testing resulted in a sensitivity of 0.82, 0.78, and 0.84, and a specificity of 0.26, 0.23, and 0.21, for BRD, NCD, and general disease, respectively. This suggests that automated milk feeders may serve as a useful preliminary tool in the detection of diseased calves. For example, producers could use feeding behavior data to identify calves requiring further inspection; however, they should not use feeding behavior data as a sole disease detection method.

Characterizing the attitudes and motivations of Ontario dairy producers toward udder health.

Considerable research has focused on identifying risk factors for intramammary infections, yet mastitis remains a pervasive disease on dairy farms. Increasingly, researchers are appreciating the role of dairy producer mindset in determining management style and thus udder health status of the herd. The objective of this study was to explore the attitudes and motivations of Ontario dairy farmers toward udder health in herds with varying bulk milk somatic cell count (BMSCC). In December 2011, 5 focus groups were conducted across Ontario, Canada, with independent groups of dairy producers representing low, medium, and high BMSCC herds. Groups were established based on producer's weighted BMSCC levels as recorded over the summer of 2011. A semi-structured interview guide was followed to discuss topics relating to udder health. Thematic analysis was performed on the interview transcripts. Generally, producers noted management techniques (specifically culling infected cows and monitoring BMSCC), a perceived wealth of information on mastitis control, and a proactive whole-herd management approach engender the perception of control over mastitis. Producers in the low BMSCC group were confident in their level of knowledge and control of mastitis in their herds, whereas high BMSCC producers generally felt lower levels of control. Several areas were identified by producers that counteract this perception, contributing to perceived low levels of control over mastitis. Participants identified that at certain times they do not understand the cause of BMSCC on their farm. This attitude was especially prominent in the high BMSCC group. Other times, producers cited improper sample handling, seasonal issues, perceived milk culture shortcomings, and low herd size as factors that limited their control over mastitis in their herds. Though producers generally have high levels of self-efficacy beliefs when it comes to udder health management, the perception still exists that, under certain situations, mastitis is uncontrollable. This highlights the fact that educational and extension efforts need to focus on ensuring that producers employ proven mastitis diagnostic, prevention, and treatment practices in a systematic manner, with realistic expectations.

Dairy farmers' expectations and receptivity regarding animal welfare advice: A focus group study.

Discussion and incorporation of best practices for animal welfare have been increasing in research, and in commercial operations, including through welfare assessment initiatives. The aim of this study was to explore dairy farmers' perceptions about being approached and receiving advice about animal welfare (i.e., lameness, hock injuries, and disbudding practices). It is useful for dairy consultants, researchers, or animal welfare assessment programs to gain an in-depth understanding of farmers' expectations when broaching the subject of animal welfare issues to facilitate communication about what can be perceived as a sensitive topic. We collected qualitative data using a focus group methodology. Five focus groups of farmers (n = 36 in total), took place in Ontario, Canada. Discussions were audio-recorded and transcribed verbatim. Participant age ranged from 21 to 80 yr (median = 47). Represented herd size ranged from 25 to 550 milking cows (median = 75). Farm type included free stall (n = 14) and tie stall (n = 22). Rigor was incorporated by using systematic thematic analysis: transcripts were coded line by line, and codes were categorized and then expanded and collapsed into themes, which were further refined to reflect farmer perceptions in a thematic map. Thematic analysis of the focus group discussions suggested 6 major themes related to farmers' receptivity to and expectations of animal welfare advice. Themes 1 to 4 provided insights into what farmers expected from those who were broaching topics: (1) an established relationship with the farmer; (2) expertise in dairy care/welfare; (3) prevention of "barn blindness"; and (4) provision of animal care services before and after welfare issues are broached. Theme 5 helped determine how welfare topics should be broached: the communication approach. Theme 6 identified who farmers feel should broach animal welfare topics on farm. Focus group discussions also provided insights into potential disconnects between farmer and veterinary expectations about animal welfare issues during herd health visits. Those who have established relationships with farmers are better received and are expected to broach welfare issues, especially if they are perceived to be an expert in animal care and welfare, and if they communicate the issue tactfully and work with the farmer to establish a plan of action.

Effect of health status upon arrival at a single milk-fed veal facility on breakeven purchase price of calves.

Male calves are purchased from the dairy industry in North America to produce red meat. The price paid for male calves varies widely, and it is unclear which variables influence the price paid for each calf. The objective of this study was to assess how the health traits of calves at the time of arrival and demographic variables affect the breakeven purchase price of a male calf entering the veal industry. A financial model was constructed using the prevalence of health abnormalities, weight at arrival, source of the calf, number of days in the barn, base carcass price, days to mortality, feed costs, season at arrival, interest rate, housing location, carcass dressing percentage, and costs associated with housing, labor, utilities, trucking, and health to calculate the breakeven purchase price and an estimate of profit. Sensitivity analysis was conducted using health variables measured at arrival and demographic variables, including season at arrival and housing location, to identify the factors with the largest impact on the predicted average daily gain, early and late mortality risk, breakeven purchase price, and profit. At the baseline inputs, the average calculated profit was -$5.36 per calf and it was most sensitive to the location of housing where calves were fed and the body weight of the arriving calf. The mortality risk in the first 21 d after arrival (early) was calculated to be 2.2%, whereas the risk of mortality after 21 d (late) was 3.7%. The risks of early and late mortality were most sensitive to the level of dehydration measured at arrival and the season at arrival for the purchased calves, respectively. The calculated average daily gain was 1.12 kg/d and it was most sensitive to housing location. The breakeven purchase price was calculated to be $242.49 per calf, which was most sensitive to the housing location where the calves were fed. The results of this analysis demonstrate that veal producers need to consider many variables before purchasing calves. In addition to overall market conditions, veal producers should factor health characteristics and the expected performance of the calves they purchase into what they are willing to pay for them.

Technical note: Assessment of neutrophil endocytosis and proteolytic degradation and its relationship with phagocytosis and oxidative burst in dairy cows.

Conventional assays of polymorphonuclear cell (PMN, neutrophil) function such as oxidative burst (OB) and phagocytosis (PC) are widely used to evaluate innate immunity in the transition period of dairy cows. Oxidative burst is commonly evaluated by measuring PMN median fluorescence intensity (MFI) involving the release of reactive oxygen species after in vitro stimulation. Phagocytosis can be measured by engulfment of fluorescent beads by PMN. DQ-ovalbumin (DQ-OVA) is a molecule suitable for the assessment of intracellular proteolytic degradation, so it might be informative about an additional pathway of pathogen handling by PMN. In this study, we evaluated the use of the DQ-OVA assay for the assessment of PMN function and the relationships among OB, PC, and DQ-OVA results in PMN isolated from blood of dairy cows between 5 and 21 d post partum. Results of the DQ-OVA validation assay were assessed with mixed linear regression models. Pearson correlation tests and kappa values for agreement were used to associate the MFI between each PMN function assay (OB, PC, and DQ-OVA). For the validation assay (9 cows in 3 replicates), PMN incubated with DQ-OVA were stimulated with IFN-γ or inhibited with cytochalasin D, and fluorescence was compared with untreated PMN. Stimulated and inhibited PMN had greater (970 ± 160 units) and lesser (593 ± 55 units) MFI relative to untreated PMN (791 ± 154 units), respectively, indicating that DQ-OVA fluorescence reflected enhanced or reduced endocytic and proteolytic function. To associate the MFI outcomes among OB, PC, and DQ-OVA, 153 samples from 40 cows were analyzed. Results showed significant, although weak association between DQ-OVA and PC MFI (Pearson r = 0.16). When values of MFI were categorized according to the first ("high" PMN functionality), second and third ("moderate" PMN functionality), or fourth ("low" PMN functionality) quartiles, agreement beyond chance (κ) was moderate: κ = 0.38 for DQ-OVA and OB, κ = 0.43 for DQ-OVA and PC, and κ = 0.43 for OB and PC. The DQ-OVA assay may complement traditional PMN functional assays because it provides additional information regarding the combination of endocytosis and proteolytic degradation, but it is not a substitute for assessment of OB or PC.