Invited review: Nutritional and management factors that influence colostrum production and composition in dairy cows.

Colostrum is a rich source of nutritional and non-nutritional components and is recognized as essential to transfer passive immunity to newborn calves. Because of the individual and seasonal variability in colostrum yield and composition, maintaining an adequate supply of high-quality colostrum year-round remains a challenge for commercial dairy producers. In this narrative review, we described the individual, seasonal, and herd-level variability of colostrum production and summarized the association between individual animal factors such as parity, sex of the calf, calf birth weight, as well as indicators of the cow's metabolic status and the yield and composition of colostrum. Further, we reviewed the current knowledge on the influence of prepartum nutrition and management strategies on colostrum production. Research on the metabolizable energy and protein supplied in the prepartum diet as well as on the inclusion and source of vitamins, minerals, and feed additives suggests prepartum nutrition influences the yield, quality, and composition of colostrum. Furthermore, the prepartum environment and dry period length remain influential factors in the production of colostrum. However, additional research is needed to understand the mechanisms by which prepartum nutrition and management affect colostrum production. Finally, time from calving to colostrum harvest and oxytocin administration as well as the current knowledge on the effect of heat treatment and colostrum storage strategies on colostral components were discussed. To conclude, we identify critical gaps in knowledge for future focus of investigation in colostrum research.

Association of uterine health in the first lactation with transition cow health and reproductive performance in the second lactation of Holstein dairy cows.

The objective of this study was to evaluate the effects of puerperal metritis (PM) diagnosed and treated during the early postpartum period of the first lactation on transition cow health, milk production, reproduction, and culling of dairy cows in their second lactation. Diagnosis of PM was based on fetid watery red-brown uterine discharge and rectal temperature above 39.5°C. Two farms were enrolled in this retrospective observational cohort study (Farm A and B). In both farms, the following diseases were recorded during the first 30 DIM in lactation 1 and 2: clinical hypocalcemia (CH), retained fetal membrane (RFM), PM, hyperketonemia (KET), left displaced abomasum (LDA) and clinical mastitis (MAST). Statistical analyses were performed using SPSS for Windows separately for each farm. Linear and logistic regression models were used for continuous (e.g., milk yield) and binary (e.g., disease, pregnancy per AI, pregnancy loss) outcomes, respectively. Cox proportional hazard regression models were calculated to model the time to event outcomes for culling or death during the first 60 DIM and for pregnancy within 250 d of the second lactation. The initial models contained the following variables: year of calving, month of calving, calving ease, stillbirth, twins, days open in lactation 1, 305 d milk yield in lactation 1, PM in lactation 1, and PM in lactation 2 as explanatory variables. A total of 4,834 cows (Farm A) and 4,238 cows (Farm B) in the second lactation were considered for statistical analyses. On farm A, the incidence of PM in lactation 1 and 2 was 20.1% and 11.2%, respectively. On farm B, the incidence of PM in lactation 1 and 2 was 14.4% and 8.5%, respectively. On both farms, cows with PM in their first lactation had greater odds for RFM and PM in their second lactation, while there was no association of PM in the first lactation with any other non-uterine diseases (i.e., CH, KET, LDA, and MAST) in the second lactation. Cows with PM in lactation 2 had reduced milk yield. The reduction in milk yield in second lactation was greater for cows that already experienced PM in lactation 1. On Farm A, cows with PM in their first lactation had a greater hazard for culling within 60 DIM of the second lactation; however, the same association was not present on Farm B. Cows with PM in lactation 1 had reduced pregnancy per AI at first service in the second lactation only on farm B. Cows with PM in lactation 2 had reduced pregnancy per AI at first service in the second lactation on both farms. Pregnancy loss in lactation 2 was only associated with PM in lactation 2 but not with PM in lactation 1. On both farms, cows had a reduced hazard for pregnancy in their second lactation within 250 DIM when they experienced PM in either lactation. In conclusion, PM in the first lactation had long-lasting negative consequences (i.e., risk of uterine disease and lower reproductive performance) for cows in their next lactation.

Effect of 2 dosages of prepartum cholecalciferol injection on blood minerals, vitamin D metabolites, and milk production in multiparous dairy cows: A randomized clinical trial.

The objective of the present study was to evaluate the effect of 2 dosages of prepartum cholecalciferol injection on blood minerals, vitamin D metabolites, and milk production. Cows entering their second or greater lactation (n = 158) were randomly assigned to a control group (CON) or one of 2 treatment groups receiving either 6 × 106 IU (6VitD) or 12 × 106 IU (12VitD) cholecalciferol intramuscularly on d 275 ± 1.2 (SD) of gestation. Concentrations of serum total Ca (tCa), phosphate, and Mg were determined on 1, 2, 3, 5, 7, and 10 d in milk (DIM). For a subsample of 30 cows entering the third lactation (n = 10/group), these samples were analyzed for cholecalciferol, 25-hydroxycholecalciferol (25-OHD3), and 24,25-dihydroxycholecalciferol (24,25-[OH]2D3). In these cows, we also determined 1,25-dihydroxycholecalciferol (1,25-[OH]2D3), the biologically most active metabolite, on 1, 2, 3, and 5 DIM. Repeated measures ANOVA was performed to evaluate the effect of different dosages of cholecalciferol on blood minerals, vitamin D metabolites, and milk yield over the first 5 test days after calving. Binary outcomes such as retained placenta and metritis were analyzed using a chi-squared test. Although the 12VitD treatment increased tCa concentrations on 1, 2, and 3 DIM compared with CON, administration of 6VitD increased tCa concentrations only on 1 DIM. Compared with CON cows and 6VitD cows, 12VitD cows had greater serum phosphate concentration during the first 10 DIM. Furthermore, 6VitD cows had greater serum phosphate concentrations compared with CON cows. On the contrary, 12VitD cows had lower serum Mg concentrations during the first 10 DIM compared with CON and 6VitD cows. Cholecalciferol was increased by the treatment and decreased quickly until 10 DIM. In respect to 25-OHD3, the 6VitD treatment resulted in a 4.1-fold increase in comparison to the CON group, while a 6.5-fold increase was observed in 12VitD animals. The vitamin D metabolite 24,25-(OH)2D3 increased linearly with 25-OHD3 serum levels, resulting in the highest concentrations in the 12VitD group. An increase of 1,25-(OH)2D3 until 3 DIM was observed in all cows. However, this rise was most pronounced in the CON group. The incidence of retained placenta was 1.9%, 11.5%, and 29.6%, and that of metritis was 11.5%, 15.4%, and 31.5% for CON, 6VitD, and 12VitD cows, respectively. Although none of the treated cows exerted clinical signs of hypocalcemia, one cow in CON incurred clinical hypocalcemia. Cows of the 12VitD group had a lower milk yield over the first 5 monthly test days compared with the control and 6VitD group (42.2 ± 0.5, 42.0, ± 0.5 and 40.7 ± 0.5 kg for control cows, 6VitD cows and 12VitD cows, respectively). Although no negative side effects were observed in 6VitD cows, we do not recommend the general application of 6 × 106 IU cholecalciferol before calving as positive effects on calcium homeostasis were marginal and restricted to the first DIM. The present findings confirm that the application of 12 × 106 IU cholecalciferol negatively affected milk production on this farm.

Association between genomic daughter pregnancy rate and expected milk production on the resumption of estrus behavior in Holstein cattle.

The objective of this observational study was to evaluate estrous expression at the first estrus occurring between 7 to 30 d in milk (DIM), as detected by an automated activity monitor (AAM), and its association with genomic daughter pregnancy rate (GDPR) and genomic expected milk production (GEM) in lactating dairy cows. A total of 4,119 lactations from 2,602 Holstein cows were included. Cows were enrolled as first lactation (n = 1,168), second lactation (n = 1,525) and third and greater lactation (n = 1,426). Hair samples were collected from the tail switch, and cows were genotyped using an SNP platform (Clarifide, Zoetis, São Paulo, SP, Brazil). Postpartum cows were examined daily by the farm personnel from calving until 10 DIM. Calving was classified as assisted (forced calf extraction) and unassisted (normal calving). Retained fetal membranes (RFM), hyperketonemia (KET), and left displaced abomasum (LDA) were also recorded. Mean GDPR (± SD) was -0.29 ± 1.4, and the intensity and duration of the first estrus event was 15.9 ± 13.1 x-factor (intensity unit measurement) and 11.1 ± 3.8 h, respectively. Cows that had greater GDPR had greater intensity and longer duration of estrus at the alert, independent of parity. Overall resumption of estrous expression, between 7 to 30 DIM, was 41.2% (1,695/4,119), where 58.8% (2,424/4,119) did not have an estrus event, 31.0% (1,274/4,119) of cows had one event of estrus, and 10.2% (421/4,119) of cows had 2 or more events of estrus early postpartum. Mean DIM (± SD) at first estrus event, detected by the AAM, was 19.4 ± 4.4 d. Days in milk at first event for cows with one event was 20.7 ± 1.6 d and 15.9 ± 3.1 d for cows with 2 or more events of estrus. First lactation cows were more likely to have an estrus event early postpartum when compared with second and third and greater lactation cows (45.2 ± 1.4% [530/1168] vs. 41.6 ± 1.3% [636/1525] vs. 37.2 ± 1.3% [529/1426], respectively). There was an interaction of parity and GDPR on the proportion of cows demonstrating an early postpartum estrus. There was no difference in the proportion of cows with an early postpartum estrus between those with assisted or unassisted calving, RFM, or LDA. However, cows that had KET were less likely to have an alert early postpartum when compared with cows that did not have KET. Mean genomic expected milk production (± SD) was 256.8 ± 600.1 kg. There was no interaction between GEM and parity on estrous expression (i.e., intensity and duration). There was no interaction between GEM and GDPR on the proportion of estrus early postpartum.

Associations between days in the close-up group and milk production, transition cow diseases, reproductive performance, culling, and behavior around calving of Holstein dairy cows.

The objective of experiment I was to evaluate the association between days in the close-up group (DINCU) and milk production, early lactation diseases, reproductive performance, and culling. In experiment II behavioral changes associated with DINCU were evaluated using a neck-mounted sensor (Smarttag neck, Nedap Livestock Management, Groenlo, the Netherlands). Cow-lactations of 28,813 animals from 14,155 individual cows of 2 farms in northern Germany and western Slovakia, calving between January 2015 and December 2020, were included in the study. After exclusion of cows with a gestation length <262 and >292 d and cows with >42 DINCU data from 8,794 and 19,598 nulliparous and parous cows, respectively, were available for final statistical analyses. To analyze the association between DINCU and second test-d 305-d mature-equivalent milk projection, linear mixed models were calculated. Binary data (i.e., clinical hypocalcemia, hyperketonemia, retained placenta [RP], acute puerperal metritis [APM], mastitis, left displaced abomasum [LDA], first service pregnancy risk) were analyzed using logistic regression models. To analyze the association between DINCU and culling or death during the first 300 DIM Cox proportional hazards were used. To analyze the association between DINCU and behavior 7 d before to 7 d after calving (i.e., activity, inactivity, eating, ruminating time), linear mixed models were calculated. Nulliparous cows with a short (<10 DINCU) and a long stay (>30 DINCU) in the close-up group had a lower milk production an increased risk for hyperketonemia, RP, and APM compared with nulliparous cows with DINCU between 21 to 28 d. Parous cows with a short (<10 DINCU) and a long stay (>30 DINCU) in the close-up group had a lower milk production, an increased risk for RP and mastitis, a reduced first service pregnancy risk, and an increased culling risk, compared with parous cows with DINCU between 21 to 28 d. Furthermore, the risk for clinical hypocalcemia and LDA was increased in parous cows with >30 DINCU compared with parous cows with <30 DINCU. The risk for APM was increased in parous cows with <10 DINCU compared with parous cows with >10 DINCU. In nulliparous cows no association was found between DINCU and the risk for left displaced abomasum and mastitis. In experiment II, cows with 7 and 35 DINCU had an impaired behavior around calving compared with cows with 14, 21, and 28 DINCU. During the last 7 d before parturition, these cows were more inactive and had a reduced eating and ruminating time. After calving, cows with 7 DINCU spent less time eating. In conclusion, cows with <10 DINCU and cows with >30 DINCU had a lower milk production, a higher risk to incur diseases and an impaired behavior, especially before calving.

Association between transfer of passive immunity, health, and performance of female dairy calves from birth to weaning.

The objective of this observational study was to compare calf health, average daily weight gain, and calf mortality considering the proposed categories of transfer of passive immunity (TPI) by the consensus report of Lombard et al. (2020). The consensus report defines 4 categories of passive immunity (excellent, good, fair, and poor) of calves obtained after colostrum ingestion. The association between the 4 TPI categories was analyzed on calf health (i.e., hazards for morbidity and mortality), and average daily weight gain (ADG) of female Holstein Friesian calves during the first 90 d of age. A further aim of this study was to examine the effects of calving-related factors, such as dystocia or winter season, on TPI status. We hypothesized that calves with excellent TPI have greater ADG, lower risks for infectious diseases such as neonatal diarrhea, pneumonia, and omphalitis, and lower mortality rates. This observational study was conducted from December 2017 to March 2021. Blood was collected from 3,434 female Holstein Friesian dairy calves from 1 commercial dairy farm. All female calves aged 2 to 7 d were assessed for TPI status by determination of total solids (TS) in serum via Brix refractometry by the farm personnel once a week. Passive immunity was categorized according to Lombard et al. (2020) with excellent (≥9.4% Brix), good (8.9-9.3% Brix), fair (8.1-8.8% Brix), or poor TPI (<8.1% Brix). For the analysis of ADG and calving ease 492 or 35 calves had to be excluded due to missing data. The distribution of calves according to TPI categories was as follows: 4.8% poor (n = 166), 29.5% fair (n = 1,012), 28.3% good (n = 971), and 37.4% excellent (n = 1,285). From the calving-related factors, parity of the dam, calving ease, birth month, calving assistance by different farm personnel, and day of life for TPI assessment were significantly associated with TS concentration. Out of 3,434 calves, 216 (6.3%) had diarrhea, and 31 (0.9%) and 957 (27.9%) suffered from omphalitis and pneumonia during the first 90 d of life, respectively. Overall, the morbidity during the preweaning period was 32.6% (n = 1,118), and the mortality was 3.1% (n = 107). The ADG was 0.90 ± 0.15 kg with a range of 0.32 to 1.52 kg. The Cox regression model showed that calves suffering from poor TPI tended toward a greater hazard risk (HR) for diarrhea (HR = 1.57, 95% CI: 0.92-2.69) compared with calves with excellent TPI. Calves suffering from TPI had a greater HR for pneumonia (HR = 2.00, CI: 1.53-2-61), overall morbidity (HR = 1.99, CI: 1.56-2.55), and mortality (HR = 2.47, CI: 1.25-4.86) in contrast to excellent TPI. Furthermore, calves with good and fair TPI had significantly greater HR for pneumonia (good TPI: HR = 1.35, CI: 1.15-1.59; fair TPI: HR = 1.41, CI: 1.20-1.65) and overall morbidity (good TPI: HR = 1.26, CI: 1.09-1.47; fair TPI: HR = 1.32, CI: 1.14-1.53) compared with the excellent TPI category. Average daily weight gain during the first 60 d of life was associated with TPI categories. Calves with excellent and good TPI status had ADG of 0.90 ± 0.01 kg/d and 0.92 ± 0.01 kg/d (mean ± SE), respectively. The ADG of calves with fair TPI status was 0.89 ± 0.01 kg/d, and calves suffering from poor TPI had 0.86 ± 0.01 kg/d. Average daily weight gain differed in calves with poor TPI compared with the other categories. Fair and excellent TPI differed additionally from good TPI. We found no statistical difference between the TPI categories fair and excellent. In conclusion, poor TPI was associated with higher morbidity and mortality during the first 90 d of life. Furthermore, calves with fair, good or excellent TPI had greater ADG.

Effect of dose and frequency of prostaglandin F2α treatments during a 7-day Ovsynch protocol with an intravaginal progesterone releasing device on luteal regression and pregnancy outcomes in lactating Holstein cows.

Our objective was to evaluate the effect of 3 different Ovsynch protocols on progesterone (P4) and pregnancies per artificial insemination (P/AI), where all cows received a P4 releasing intravaginal device (PRID) from d 0 until d 8. We hypothesized that (1) both modified PGF2α treatments lead to decreased P4 at the second GnRH treatment (G2), resulting in greater P/AI, (2) the treatment effect is influenced by the presence of a corpus luteum (CL) at the beginning of the protocol, and (3) potential vaginal discharge caused by the PRID does not have a negative influence on fertility. Lactating Holstein cows (n = 1,056) were randomly assigned to 1 of 3 treatment groups on a weekly basis (n = 356; control: d 0, 100 µg of GnRH + PRID; d 7, 25 mg of dinoprost; d 8, PRID removal; d 9, 100 µg of GnRH). Cows in the second group (n = 353) received an Ovsynch protocol with a double dose of PGF2α (DoubleDose: d 0, 100 µg of GnRH + PRID; d 7, 50 mg of dinoprost; d 8, PRID removal; d 9, 100 µg of GnRH). Cows in the third group (n = 347) received an Ovsynch protocol with a second PGF2α treatment 24 h after the first one (2PGF: d 0, 100 µg of GnRH + PRID; d 7, 25 of mg dinoprost; d 8, 25 mg of dinoprost and PRID removal; d 9, 100 µg of GnRH). All cows had their ovaries scanned to determine the presence of a CL at the beginning of the Ovsynch protocol. Vaginal discharge score (VS) was evaluated at PRID removal. All cows received timed artificial insemination approximately 16 h after G2. Pregnancy diagnosis was performed via transrectal ultrasonography (d 38 ± 3 after timed artificial insemination) and rechecked on d 80 ± 7 after timed artificial insemination. Blood samples were collected on d 0, 7, and 9 of the protocol to determine P4 concentrations. Treatment affected P4 at G2. Progesterone was lower for 2PGF and DoubleDose cows compared with cows in the control group (control 0.35 ± 0.02 ng/mL; DoubleDose 0.29 ± 0.02 ng/mL; 2PGF 0.30 ± 0.02 ng/mL). Overall, P/AI did not differ among treatments. We found, however, an interaction between treatment and CL at the first GnRH treatment. Cows lacking a CL at the first GnRH treatment in the 2PGF group had greater P/AI (47.9%) compared with the same type of cows in the DoubleDose group (32.7%). We observed an effect of VS on P4 concentration at d 7. We found an increase in P4 with greater VS. Vaginal discharge score at PRID removal tended to have a positive effect on P/AI at d 38 (VS0: 36.5%; VS1: 41.3%; VS2: 49.7%). In conclusion, the addition of a second PGF treatment on d 7 and 8 of a 7-d Ovsynch protocol increased luteal regression and decreased mean P4 at G2. Cows treated with PGF2α 2 times 24 h apart showed greater P/AI, compared with cows treated with an increased dose of PGF2α.

Association of activity and subsequent fertility of dairy cows after spontaneous estrus or timed artificial insemination.

The objective of this observational study was to evaluate the association between increased physical activity at first artificial insemination (AI) and subsequent pregnancy per AI (P/AI) in lactating Holstein cows following spontaneous estrus or a timed AI (TAI) protocol. We also wanted to identify factors associated with the intensity of activity increase (PA) captured by automated activity monitors (AAM) and fertility. Two experiments were conducted, in which cows either were inseminated based on the alert of the AAM system (AAM cows) or received TAI following a 7-d Ovsynch protocol (TAI cows) if not inseminated within a farm-specific period after calving. Experiment 1 included 2,698 AI services from AAM cows and 1,042 AI services from TAI cows equipped with the Smarttag Neck (Nedap Livestock Management) from a dairy farm in Slovakia (farm 1). In the second experiment, 6,517 AI services from AAM cows and 1,226 AI services from TAI cows fitted with Heatime (Heatime Pro; SCR Engineers Ltd.) from 8 dairy farms in Germany (farms 2-9) were included. Pregnancy diagnosis was performed on a weekly basis by transrectal ultrasound (farms 1, 3, 7, 8) or by transrectal palpation (farms 2, 4-6, 9). Estrous intensity was represented by the peak value of the change in activity. In experiment 1, PA was categorized into low (x-factor 0-20) and high (x-factor 21-100) PA, and in experiment 2 into low (activity change = 35-89) and high (activity change = 90-100) PA. In TAI cows from both experiments, PA was additionally categorized into cows with no AAM alert. Data were analyzed separately for AAM and TAI cows using multinomial logistic regression models for PA in TAI cows and logistic regression models for PA in AAM cows and P/AI in both groups. In experiment 1, P/AI of AAM cows was greater for AI services performed with conventional frozen semen (57.6%) compared with sexed semen (47.2%), whereas type of semen only tended to be associated with P/AI in TAI cows (54.4% conventional frozen semen vs. 48.9% sexed semen). In experiment 2, P/AI was greater for fresh semen (AAM cows: 44.4% vs. TAI cows: 44.2%) compared with conventional frozen semen (AAM cows: 37.6% vs. TAI cows: 34.6%). In both experiments, pregnancy outcomes were associated with PA. In experiment 1, AAM cows with high PA (55.1%) had greater P/AI than cows with low PA (49.8%). Within TAI cows, cows with no alert (38.8%) had reduced P/AI compared with cows with low (54.2%) or high PA (61.8%). In experiment 2, AAM cows with high PA (45.8%) had greater P/AI compared with cows with low PA (36.4%). Timed AI cows with no alert (27.4%) had decreased P/AI compared with cows with low (41.1%) or high (50.8%) PA. The greatest risk factors for high PA were parity (experiment 1) and season of AI (except for TAI cows from experiment 1). We conclude that high PA at the time of AI is associated with greater odds of pregnancy for both AAM and TAI cows. In both experiments, about 2 thirds of AAM cows (experiment 1: 69.9% and experiment 2: 70.7%) reached high PA, whereas only approximately one-third or less of TAI cows (experiment 1: 37.3% and experiment 2: 23.6%) showed high PA. Although we observed similar results using 2 different AAM systems for the most part, risk factors for high PA might differ between farms and insemination type (i.e., AAM vs. TAI).

Effect of transition cow health and estrous expression detected by an automated activity monitoring system within 60 days in milk on reproductive performance of lactating Holstein cows.

The objective of this observational study was to evaluate the association of transition cow health and estrous expression, detected by an automated activity monitoring system (Smarttag Neck, Nedap Livestock Management), with reproductive performance in lactating Holstein cows. A total of 3,750 lactating Holstein cows (1,563 primiparous cows and 2,187 multiparous cows) from a commercial dairy farm in Slovakia calving from January 2020 until July 2021 were enrolled on an ongoing basis. Activity data were recorded from d 7 until d 60 postpartum. Within this observational period, cows were classified into 3 categories: (1) no estrus event (Estrus0), (2) 1 estrus event (Estrus1), or (3) 2 or more estrus events (Estrus2+). Transition cow health was assessed by farm personnel within the first 30 d in milk (DIM) using standard operating procedures. Generalized linear mixed models were used to analyze continuous and categorical data. Cox proportional hazard models were used for time to event data. The overall prevalence of anestrus was 20.8%. Multiparous cows had a greater risk for anestrus compared with primiparous cows [odds ratio (OR) = 1.4]. Cows with stillbirth (OR = 1.76), retained placenta (OR = 2.19), puerperal metritis (OR = 1.48), or subclinical ketosis (OR = 1.51) had a greater risk for anestrus. In addition, cows calving in summer (OR = 0.82), autumn (OR = 0.38), or winter (OR = 0.56) had a higher incidence of anestrus than cows calving in spring. Estrous expression from d 7 until d 60 postpartum was associated with estrous duration (DU) and estrous intensity at first artificial insemination (AI). Cows in Estrus0 had the shortest DU at first postpartum AI (9.4 ± 0.18 h) compared with cows in Estrus1 (10.5 ± 0.13 h) and Estrus2+ (11.4 ± 0.12 h). Cows in Estrus2+ had a longer DU at first postpartum AI compared with cows in Estrus1. For Estrus0, Estrus1, and Estrus2+ cows, pregnancy per AI at first service was 42.5%, 50.9%, and 55.4%, respectively. Estrous expression from d 7 until d 60 postpartum was associated with time to first AI and time to pregnancy. Compared with Estrus0 cows, Estrus1 [hazard ratio (HR) = 1.43] and Estrus2+ cows (HR = 1.62) had an increased hazard of being inseminated within 100 DIM. Compared with Estrus2+, Estrus1 cows had a reduced hazard of being inseminated within 100 DIM (HR = 0.89). Compared with Estrus0 cows, Estrus1 (HR = 1.24) and Estrus2+ cows (HR = 1.46) had an increased hazard of becoming pregnant within 200 DIM. Median DIM to pregnancy were 121, 96, and 92 for Estrus0, Estrus1, and Estrus2+ cows, respectively. In conclusion, cows with transition cow disorders (i.e., stillbirth, retained placenta, puerperal metritis, or subclinical ketosis) had a greater chance for anestrus compared with healthy cows. Cows in Estrus0 had reduced estrous expression at first AI and inferior reproductive performance compared with cows that displayed estrous activity from d 7 until d 60.

Evaluation of laboratory and on-farm tests to estimate colostrum quality for dairy cows.

The objectives of this study were to evaluate different analytical methods to determine colostrum quality in dairy cattle, including one laboratory-based method (ELISA) and 4 on-farm tests. We hypothesized that the colostral IgG concentration using different analytical methods, such as ELISA (mg/mL), digital Brix refractometer (% Brix), colostrometer (specific gravity and mg/mL), an outflow funnel (seconds), and a lateral flow assay (mg/mL), were highly correlated with the reference method, radial immunodiffusion (RID; mg/mL) and would generate comparable results. Colostrum samples were collected from 209 Holstein Friesian cows on 2 commercial dairy farms in Germany. Colostrum weight and colostrum temperature were measured. Test characteristics, such as optimum thresholds, sensitivity, specificity, and area under the curve (AUC) were determined using a receiver operating characteristic curve analyses for each test. Out of 209 colostrum samples assessed by RID, 186 (89%) samples had high quality (≥50 mg IgG/mL), while 23 colostrum samples (11%) showed poor quality with IgG concentrations less than 50 mg/mL. The mean IgG concentration (±SD) was 101.3 ± 45.9 mg/mL and the range was 6.0 to 244.3 mg/mL. The Pearson correlation coefficient (r) between RID and ELISA was r = 0.78. In comparison to RID, Pearson correlation coefficients for the on-farm tests were: r = 0.79 (digital Brix refractometry), r = 0.58 (colostrometer: specific gravity), r = 0.61 (colostrometer: temperature corrected), r = 0.26 (outflow funnel) and r = 0.43 (lateral flow assay), respectively. The optimal threshold to identify high-quality colostrum using ELISA was 50.8 mg/mL with sensitivity 91.3%, specificity 92.3%, and AUC of 0.94. For the on-farm tests sensitivity ranged from 95.7% (Brix refractometry) to 60.9% (lateral flow assay). Specificity ranged from 88.6% (lateral flow assay) to 75.9% (colostrometer: temperature corrected). The AUC ranged from 0.93 (Brix refractometry) to 0.73 (outflow funnel). Based on the AUC, ELISA (0.94) and Brix refractometry (0.93) can be considered highly accurate. In conclusion, the ELISA is accurate to assess colostrum quality. Regarding the on-farm tests only the digital Brix refractometer and the colostrometer were adequate to determine colostrum quality.

Effects of a single transdermal administration of flunixin meglumine in early postpartum Holstein Friesian dairy cows: Part 1. Inflammatory and metabolic markers, uterine health, and indicators of pain.

The objectives of this study were to assess the effects of a single transdermal administration of flunixin meglumine (FM) in early postpartum Holstein Friesian dairy cows on serum concentrations of inflammatory and metabolic markers, uterine health, and indicators of pain. The hypothesis was that the anti-inflammatory, antipyretic, and analgetic effects of the pharmaceutic agent would reduce systemic inflammation, resulting in improved metabolic and inflammatory profile, diminished incidence of metritis, and reduced expression of pain. A total of 500 cows (153 primiparous, 347 multiparous) from 3 different commercial dairy farms in the northeast of Germany were included in a randomized controlled clinical trial. Farms were preselected based on high haptoglobin concentrations in their fresh lactating cows. Cows were excluded if they had experienced dystocia, stillbirth, or twin birth, or if they showed any signs of milk fever, retained fetal membranes, or fever (>40°C). The cows were treated once with either FM (3.33 mg/kg) or a placebo as control (CON) through transdermal administration between 24 to 36 h postpartum (d 2). General health examinations were performed (daily from d 2-8 and additionally on d 15 postpartum), vaginal discharge was assessed using the Metricheck device (d 8 and 15 postpartum) and serum samples were analyzed for inflammatory and metabolic markers (d 2, 4, and 6 postpartum). Effects of treatment, parity, sampling day, and their interactions were evaluated using mixed effects models. Primiparous cows treated with FM showed lower serum haptoglobin concentrations (0.90 ± 0.08 vs. 1.17 ± 0.07 g/L; ± standard error of the mean) and higher serum albumin concentrations (35.5 ± 0.31 vs. 34.8 ± 0.31 g/L) on d 6 postpartum. They also had a lower risk for purulent vaginal discharge with or without a fever compared with CON cows on d 15 postpartum (odds ratio for CON vs. FM: 1.63, 95% CI: 1.26-2.00), and body temperature was lower throughout the first 15 d in milk (39.1 ± 0.11 vs. 39.2 ± 0.11°C). Multiparous cows treated with FM had lower serum ß-hydroxybutyrate concentrations on d 4 postpartum (0.71 ± 0.05 vs. 0.78 ± 0.05 mmol/L) and d 6 postpartum (0.74 ± 0.05 vs. 0.80 ± 0.05 mmol/L). Regardless of parity, FM-treated cows were significantly less likely to abduct their tail from their body (14.3 vs. 23.6%) and show an arched back (27.9 vs. 39.7%) on the day after treatment compared with CON cows. It can be concluded that FM treatment slightly reduced inflammation and diminished the risk for metritis in primiparous cows, improved metabolic profile in multiparous cows, and reduced expressions of pain in all cows.

Effects of a single transdermal administration of flunixin meglumine in early postpartum Holstein Friesian dairy cows: Part 2. Milk yield, culling risk, and reproductive performance.

This study was conducted to assess the effects of a single transdermal administration of flunixin meglumine (FM) in early postpartum Holstein Friesian dairy cows on milk yield, culling risk, and reproductive performance. We hypothesized that FM treatment would reduce systemic inflammation, leading to higher milk yield, reduced culling risk, and better reproductive performance in the subsequent lactation. Holstein Friesian dairy cows [n = 500, 153 primiparous (PRIM), 347 multiparous (MULT)] from 3 farms in northeast Germany were enrolled in a prospective, randomized controlled clinical trial. Farms at risk for cows with excessive postpartum inflammation were identified in a preliminary trial by measuring serum haptoglobin concentrations in their fresh lactating cows. Only cows that had a eutocic birth and delivered a singleton calf alive, with no signs of milk fever or retained fetal membranes and rectal temperature ≤40°C at first clinical examination, were included within 24 to 36 h postpartum. Treatment included a single transdermal administration of either FM (3.33 mg/kg) or a placebo as control (CON). Milk production, milk solids, urea, and somatic cell count were recorded monthly for 8 mo after calving. Culling risk, first-service conception risk, and days open were retrieved from the farms' herd management software. Separate models for PRIM and MULT cows were built for most parameters because of significant effects of parity and parity × treatment interaction. Energy-corrected milk yield from 8 monthly Dairy Herd Improvement-equivalent tests was slightly greater in PRIM cows treated with FM (29.51 and 30.73 ± 1.35 kg, CON vs. FM), whereas it was reduced in treated MULT cows (38.23 and 37.47 ± 1.17 kg, CON vs. FM) compared with CON. Milk fat and protein yields were greater in FM-treated PRIM cows and lower in treated MULT cows compared with CON. Milk urea and somatic cell count were not affected by treatment. No differences in culling risk, first-service conception risk, or days open were observed. We conclude that a single transdermal administration of FM in early postpartum dairy cows on farms at risk for excessive postpartum inflammation slightly increased milk, milk fat, and milk protein yields in PRIM cows and decreased these variables in MULT cows. Neither culling risk nor fertility was affected by treatment in this study.

Management-related factors in dry cows and their associations with colostrum quantity and quality on a large commercial dairy farm.

The objective of this observational study was to evaluate the association of management-related factors in dry cows and colostrum quantity and quality in Holstein cows on a large commercial dairy farm. This study was conducted from January 2018 to December 2020 on a commercial dairy farm in Germany, milking approximately 2,500 Holstein cows. Dairy personnel recorded colostrum quantity (n = 7,567) and evaluated colostrum quality in a subsample of animals (n = 2,600) using a digital Brix refractometer. Generalized linear mixed models were constructed to evaluate the association of management-related factors and colostrum quantity and quality. Models were run separately for primiparous or multiparous cows. The outcome variable was either colostrum quantity (kg) or quality (% Brix). Average colostrum quantity was 4.0 ± 2.5 kg, 5.1 ± 3.4 kg, and 5.5 ± 3.5 kg for cows in lactation 1, 2, and ≥3, respectively. In primiparous cows (n = 2,351), colostrum quantity was affected by month of calving (greatest in April = 4.1 kg, and lowest in November = 3.2 kg), sex of the calf (female singleton = 3.50 ± 0.26 kg; male singleton = 3.76 ± 0.27 kg; twins = 2.97 ± 0.66 kg), stillbirth (stillbirth = 3.14 ± 0.39 kg; no stillbirth = 3.68 ± 0.31 kg). In multiparous cows (n = 5,216), colostrum quantity was affected by month of calving (greatest in May = 5.5 kg, and lowest in October = 3.8 kg), calving ease (calving ease 0 = 4.23 ± 0.26 kg; score 1 = 4.77 ± 0.21 kg; score 2 = 4.98 ± 0.22 kg; score 3 = 5.30 ± 0.22 kg), sex of the calf (female singleton = 4.42 ± 0.21 kg; male singleton = 5.00 ± 0.21 kg; twins = 5.03 ± 0.30 kg), stillbirth (stillbirth = 4.24 ± 0.38 kg; no stillbirth = 5.39 ± 0.11 kg), milk yield in previous lactation (+0.1 kg increase for 1,000 kg more milk yield in previous lactation), days spent in the far-off group (0.05 ± 0.003 kg for every day), and days in the close-up pen (0.06 ± 0.010 kg for every day). Average colostrum quality was 25.1 ± 3.4% Brix, 24.7 ± 3.3% Brix, and 27.6 ± 4.4% Brix for cows in lactation 1, 2, and ≥3, respectively. In primiparous cows (n = 817), colostrum quality was affected only by month of calving. Colostrum quality in primiparous cows was greatest in December (26.8% Brix) and lowest in August (23.9% Brix). In multiparous cows (n = 1,783), colostrum quality was affected by parity (lactation 2 = 25.2 ± 2.7% Brix; lactation 3+ = 27.9 ± 2.7% Brix), month of calving (greatest in February = 27.5% Brix, and lowest in August = 25.7% Brix), milk yield in previous lactation, and colostrum quantity. We observed a seasonal pattern for colostrum quantity and quality. Future intervention studies using multiple farms need to elucidate whether management of the photoperiod or length of exposure to close-up diets, or both, can help to optimize colostrum production.

Associations between time in the close-up group and milk yield, milk components, reproductive performance, and culling of Holstein dairy cows fed acidogenic diets: A multisite study.

The objective of this prospective cohort study was to evaluate the association between the prepartum days in the close-up group (DINCU) and milk yield, milk components, reproductive performance, and culling risk in the subsequent lactation for Holstein dairy cows. Dry cow feeding management of 20 farms was evaluated during 2 farm visits. All farms were feeding an acidogenic diet in the close-up group. Data from 14,843 cows were collected for 365 d following the second farm visit. Data sets of 13,314 cows were available for final statistical analysis after exclusion of cows with missing information about gestation length, cows with a gestation length shorter than 262 d or longer than 292, cows with 0 DINCU, and cows with >42 DINCU. At enrollment, 3,871 and 9,443 of those animals were nulliparous and parous cows, respectively. Continuous data such as energy corrected milk (ECM), the ratio of fat and protein, and somatic cell score (SCS) at first test day were analyzed using linear mixed models. Binary data such as stillbirth, culling within 60 DIM, and pregnancy within 150 DIM were analyzed using logistic regression models. Based on their different physiology, separate models were built for nulliparous and parous cows. All results displayed are the predicted least squares means from the multivariable analyses. A significant association between DINCU and milk yield at first test day was observed for nulliparous and parous cows. Nulliparous cows with 7, 21, or 35 DINCU had a first test day ECM of 31.8, 33.3, and 35.5 kg, respectively. Parous cows with 7, 21, or 35 DINCU had a first test day ECM of 42.8, 45.6, and 44.6 kg of ECM, respectively. In nulliparous cows, there was a tendency for an association between DINCU and the ratio of fat and protein at first test day. In parous cows, however, a significant association was observed. Parous cows with 7, 21, or 35 DINCU had a ratio of fat and protein of 1.31, 1.35, and 1.37, respectively. There was a significant association between DINCU and SCS at first test day in nulliparous and parous cows. In nulliparous cows with 7, 21, or 35 DINCU, SCS was 2.39, 2.49, and 2.85, respectively. In parous cows with 7, 21, or 35 DINCU, SCS was 2.46, 2.53, and 2.78, respectively. No associations were observed between DINCU and occurrence of stillbirth and DINCU and the risk of pregnancy within 150 DIM. The multivariable model predicted a tendency for an association between DINCU and the risk of being culled within 60 DIM in parous cows. Particularly, 0 to 6 DINCU were associated with a substantially increased risk of being culled. In conclusion, a short stay in the close-up group should be avoided to improve milk yield at first test day and to minimize culling risk for parous cows. A long stay in the close-up group (>30 d) was associated with reduced milk production and an increased ratio of fat and protein in milk of parous cows and increased SCS of nulliparous and parous cows.

The effect of transdermal flunixin meglumine on blood cortisol levels in dairy calves after cautery disbudding with local anesthesia.

The objective of this study was to evaluate the effect of the nonsteroidal anti-inflammatory drug transdermal flunixin meglumine (Finadyne Transdermal) on plasma cortisol, average daily weight gain, and standing and lying behavior of calves, when given at the time of disbudding combined with local anesthesia. A sedative was not used to minimize pharmacological interactions. Seventy-one female Holstein Friesian calves aged 13 ± 2 d, with an average weight of 48.9 ± 4.26 kg were enrolled in the study. All calves were randomly assigned to one of 3 treatment groups: (1) control group (CON, n = 27), (2) 1-flunixin group (1-FLU, n = 26) with a single administration of transdermal flunixin meglumine at disbudding, and (3) 2-flunixin group (2-FLU, n = 24) with 2 administrations of transdermal flunixin meglumine, the first treatment at disbudding and the second 6 h after disbudding. Although the CON group received a placebo, 1-FLU and 2-FLU received flunixin meglumine transdermally. To account for plasma cortisol changes due to manipulation and handling of the calves, a sham disbudding procedure was performed one week before disbudding took place. Sham disbudding was conducted by using a cold cautery dehorner applied to each horn bud for 10 s. Disbudding was performed in a similar way by using a hot cautery dehorner. Plasma samples were collected to measure the stress biomarker cortisol at 7 different time points. Body weights were measured 4 times in 2 wk. Standing and lying behavior was assessed via 3-dimensional accelerometer. During sham disbudding and disbudding mean plasma cortisol concentrations were 6.09 ± 2.5 ng/mL and 5.16 ± 2.8 ng/mL, respectively. Treatment tended to have an effect on plasma cortisol concentrations during sham disbudding and had an effect on plasma cortisol concentrations during disbudding. Plasma cortisol concentrations were affected by treatment 2 h after disbudding in comparison to CON group. Furthermore, there was a significant effect on plasma cortisol concentrations 6 h after disbudding in contrast to CON. A return to baseline plasma cortisol levels (initial concentrations) was not achieved in CON during disbudding. There was no statistical difference between average daily weight gain and the treatment procedure. Total lying time was not affected by treatment after disbudding. In conclusion, transdermal flunixin meglumine given at the time of disbudding combined with local anesthesia decreased concentrations of the stress biomarker cortisol, but a second dose 6 h after disbudding had no further effect on plasma cortisol levels.

Randomized clinical trial to evaluate the effects of a prepartum cholecalciferol injection on postpartum serum calcium dynamics and health and performance in early-lactation multiparous dairy cows.

The objectives of the present study were (1) to evaluate the effect of prepartum cholecalciferol treatment on serum Ca concentration during the first 10 d after calving and (2) to evaluate the effect of treatment on subsequent health and performance. Multiparous Holstein cows (n = 377) from one dairy farm were fed a negative dietary cation-anion difference diet (-31 mEq/kg of DM) for the last 21 d of gestation. On d 275, the animals were randomly assigned to a control or a treatment group. Cows in the control group were left untreated, and cows in the treatment group received an injection of 12 × 106 IU of cholecalciferol intramuscularly on the day of enrollment. If treated cows did not deliver the calf within 6 d, they were reinjected with 10 × 106 IU of cholecalciferol. Blood samples were drawn on 1, 2, 3, 5, 7, and 10 days in milk (DIM) and analyzed for serum Ca, P, and Mg concentrations. In a subsample of cows (50 control cows, 35 cows treated once with cholecalciferol, and 15 cows treated twice) serum haptoglobin, nonesterified fatty acids, ß-hydroxybutyrate, and 25-hydroxycholecalciferol concentrations were analyzed on 1, 5, and 10 DIM. Binary data [retained placenta (RP), metritis] were analyzed using logistic regression models. Repeated measures ANOVA with first-order autoregressive covariance was performed to evaluate the treatment effect on milk yield over the first 10 test days after parturition, 25-hydroxycholecalciferol, serum Ca, P, Mg, ß-hydroxybutyrate, nonesterified fatty acids, and haptoglobin concentrations. Cox proportional hazards were used to model the time to event outcomes (time to pregnancy within 200 d, culling until 300 DIM). After enrollment of 31.4% of cows and a preliminary analysis, adverse reactions became apparent, and the study was stopped. Cows treated with cholecalciferol had a greater risk of incurring RP and metritis. The adjusted mean incidences were 2.0%, 7.7%, and 4.0% for RP, and 21.6%, 39.3%, and 33.3% for metritis for control cows, cows treated once, and cows treated twice with cholecalciferol, respectively. Compared with control cows, cows injected once with 12 × 106 IU of cholecalciferol produced less energy-corrected milk on the first (-3.76 kg) and second (-2.75 kg) test days, respectively. Cows injected twice with cholecalciferol (12 × 106 IU of cholecalciferol and 10 × 106 IU 1 wk later) had a reduced milk yield only at first test day (-3.80 kg). Treatment with cholecalciferol led to a significant increase in 25-hydroxycholecalciferol on d 1, 5, and 10 after calving. Serum Ca and P concentrations were significantly increased in cows treated with cholecalciferol, but serum Mg concentrations were significantly reduced. Haptoglobin concentrations were significantly increased on 5 DIM in cows injected once with 12 × 106 IU of cholecalciferol. Although we observed no effect of treatment on culling until 300 DIM, time to pregnancy was delayed by 34 d in cows injected once with 12 × 106 IU of cholecalciferol. In the present study, injection with 12 × 106 IU of cholecalciferol had detrimental effects on health and milk production despite the beneficial effects on Ca homeostasis.

Occurrence and greater intensity of estrus in recipient lactating dairy cows improve pregnancy per embryo transfer.

The aim of this study was to determine the association between occurrence and intensity of estrous expression with pregnancy success in recipient lactating dairy cows subjected to embryo transfer (ET). Two observational studies were conducted. Holstein cows were synchronized using the same timed ET protocol, based on estradiol and progesterone in both experiments. At 9 d after the end of the timed ET protocol only animals that had ovulated were implanted with a 7-d embryo [experiment 1 (Exp. 1); n = 1,401 ET events from 1,045 cows, and experiment 2 (Exp. 2); n = 1,147 ET events from 657 cows]. Embryos were produced in vivo (Exp. 1 and Exp. 2) and in vitro (only Exp. 2), then transferred to recipient cows as fresh or frozen-thawed. Pregnancy was confirmed at 29 and 58 d after the end of timed ET protocol. In Exp. 1, animals had their estrous expression monitored through a tail chalk applied on the tail head of the cows and evaluated daily for chalk removal (no estrus: 100% of chalk remaining; estrus: <50% of chalk remaining). In Exp. 2, cows were continuously monitored by a leg-mounted automated activity monitor. Estrous expression was quantified using the relative increase in physical activity at estrus in relation to the days before estrus. Estrous expression was classified as no estrus [<100% relative increase in activity (RI)], weak intensity (100-299% RI), and strong intensity (≥300% RI). Data were analyzed by analysis of variance using mixed linear regression models (GLIMMIX) in SAS (SAS Institute Inc.). A total of 65.2% (914/1,401) and 89.2% (1,019/1,142) of cows from Exp. 1 and Exp. 2, respectively, displayed estrus at the end of the ovulation synchronization protocol. In Exp. 1, cows expressing estrus before to ET had greater pregnancy per ET than those that did not [41.0 ± 2.3% (381/914) vs. 31.5 ± 2.9% (151/487), respectively]. Similarly, in Exp. 2, cows classified in the strong intensity group had greater pregnancy per ET compared with cows in the weak intensity and no estrus groups [41.3 ± 2.2% (213/571) vs. 32.7 ± 2.7% (115/353) vs. 11.3 ± 3.5% (26/218), respectively]. There was no effect of ET type on pregnancy per ET in Exp. 1. However, in Exp. 2, cows that received an in vivo-produced embryo, either fresh or frozen, had greater pregnancy per ET compared with cows that received in vitro-produced embryo. Cows receiving embryos in the early blastocyst and blastocyst stage had greater fertility compared with cows receiving embryos in the morula stage. There was an interaction between the occurrence of estrus and the stage of embryo development on pregnancy per ET, cows which displayed estrus and received a morula or early blastocyst had greater pregnancy per ET than cows that did not display estrus. In conclusion, the occurrence and the intensity of estrous expression improved pregnancy per ET in recipient lactating dairy cows and thus could be used as a tool to assist in the decision making of reproduction strategies in dairy farms.

Economic impact of adding a second prostaglandin F2α treatment during an Ovsynch protocol using a meta-analytical assessment and a stochastic simulation model.

Incomplete luteal regression after treatment with a single dose of PGF2α during an Ovsynch protocol decreases fertility to timed artificial insemination (AI). An additional treatment with PGF2α 24 h after the first dose has been recommended to increase the proportion of cows with complete luteal regression and subsequent pregnancy per AI (P/AI). This is, however, associated with additional costs of labor and product. The objective was to develop a stochastic partial budget model to estimate the economic impact of an additional PGF2α treatment on d 8 during an Ovsynch protocol in primiparous and multiparous cows. A systematic review of the literature and a meta-analytical assessment was performed to evaluate the effects of adding a second PGF2α treatment during the Ovsynch protocol on P/AI in lactating dairy cows. Thirteen randomized controlled experiments from 11 published manuscripts including 9,735 cows were used. We were able to retrieve information regarding parity from 9 experiments from 7 manuscripts (2,367 primiparous cows and 5,356 multiparous cows). An additional dose of PGF2α yielded a 5.60 risk difference in P/AI [95% confidence interval (CI) = 3.69-7.52]. In primiparous cows and multiparous cows, an additional dose of PGF2α yielded a 4.24 (95% CI = 0.31-8.17) and a 5.31 risk difference in P/AI (95% CI = 2.75-7.87). Revenue was based on the associated improvement in reproductive performance because of an increase in P/AI multiplied by the value of a pregnancy (PGVAL). Median PGVAL was €252, ranging from €42 (fifth percentile) to €623 (95th percentile). Based on parity, median PGVAL was €205 (5th percentile = €43; 95th percentile = €651) and €264 (5th percentile = €88; 95th percentile = €598) for primiparous (n = 1,252) and multiparous cows (n = 3,003), respectively. Using a stochastic simulation model with 10,000 iterations, adding a second PGF2α dose on d 8 was more profitable (€7.76/cow; 5th percentile = €0.01; 95th percentile = €29.40) compared with a single PGF2α administration on d 7. In primiparous cows, adding a second PGF2α treatment was more profitable (€1.99/cow; 5th percentile = -€3.08; 95th percentile = €22.52) in 67% of all simulated iterations. In multiparous cows, adding a second PGF2α treatment was more profitable (€7.92/cow; 5th percentile = -€0.09; 95th percentile = €28.22) in 95% of all simulated iterations. In conclusion, there was a clear benefit of an additional PGF2α treatment during the Ovsynch protocol on P/AI (+5.6 percentage units). Despite additional costs for hormones and labor, an additional treatment with PGF2α on d 8 was more profitable in 95% of all scenarios because of the associated increase in fertility. It turned out, however, that adding a second PGF2α treatment was more profitable in multiparous cows compared with primiparous cows.

Association between serum calcium dynamics around parturition and common postpartum diseases in dairy cows.

The objective of this study was to compare periparturient serum Ca dynamics (CaDyn) in cows with and without diseases in early lactation. The study enrolled 1,949 cows from a commercial dairy farm in northern Germany. Blood samples were drawn 7 d before expected calving date and on d 0, 1, 3, and 7 after calving and analyzed for serum Ca concentration. Cows were monitored for clinical hypocalcemia (CH), ketosis, left displaced abomasum (LDA), retained placenta, acute puerperal metritis (APM), mastitis, and pneumonia. To evaluate the association between CaDyn and diseases during the transition period, repeated measures ANOVA with first-order autoregressive covariance were performed. Serum CaDyn of healthy cows (i.e., without any of the aforementioned diseases) was compared with CaDyn of cows with one of the aforementioned diseases (CH, ketosis, APM, mastitis, LDA, and pneumonia), and cows with multiple diseases (CH+, ketosis+, APM+, mastitis+, LDA+, and pneumonia+). Separate models were built for primiparous and multiparous cows. For primiparous cows, we evaluated the association between CaDyn and ketosis (healthy cows vs. cows with ketosis vs. cows with ketosis+) and CaDyn and APM (healthy cows vs. cows with APM vs. cows with APM+). The same models were built for multiparous cows. Three additional models were built for multiparous cows to evaluate the association between CaDyn and CH (healthy cows vs. cows with CH vs. cows with CH+), mastitis (healthy cows vs. cows with mastitis vs. cows with mastitis+), or LDA (healthy cows vs. cows with LDA vs. cows with LDA+). In primiparous cows, serum Ca concentrations of cows with ketosis, APM, and APM+ were significantly reduced on d 3 and 7 after calving, compared with healthy cows. Serum Ca concentrations of primiparous cows with ketosis+ were reduced on d 3, but not on d 7 after calving. Multiparous cows with CH had significantly reduced serum Ca concentrations on d 0, 1, and 3 compared with healthy cows. On d 3 and 7, serum Ca concentration of CH+ cows was significantly reduced compared with healthy multiparous cows. Multiparous cows with ketosis and ketosis+ had significantly reduced serum Ca concentrations on d 1 and 3 compared with healthy cows. Cows with APM+ had significantly increased serum Ca concentrations on d 0 and reduced serum Ca concentrations on d 3, compared with healthy cows. Whereas multiparous cows with mastitis had a reduced serum Ca concentration on d 1, mastitis+ cows had a reduced serum Ca concentration on d 1 and 3, compared with healthy multiparous cows. Overall, multiparous cows with LDA+ had reduced serum Ca concentrations. Especially a delayed onset of hypocalcemia (d 3 and 7) was indicative for the development of disease in primiparous cows. In multiparous cows, reduced serum Ca concentrations on d 1 and 3 were associated with occurrence of diseases. Future studies should evaluate whether reduced serum Ca concentrations are a cause or concomitant circumstance of diseases in early lactation.

Association of estrous expression detected by an automated activity monitoring system within 40 days in milk and reproductive performance of lactating Holstein cows.

The objective of this observational study was to evaluate the association of estrous expression within 40 days in milk (DIM) using a neck-mounted automated activity monitor (Heatime Pro; SCR Engineers Ltd.) with reproductive performance in lactating Holstein cows. A total of 2,077 cows (614 primiparous cows and 1,463 multiparous cows) from 5 commercial dairy farms were included in the statistical analyses. Activity data from the first 7 d after calving were excluded. An estrus event was defined as an activity change index ≥35 for more than 2 h. Cows were classified according to the number of estrus events from d 7 until d 40 postpartum into 3 categories: (1) no estrus event (Estrus0); (2) one estrus event (Estrus1), and (3) 2 or more estrus events (Estrus2). Generalized linear mixed models were used to analyze continuous and categorical data. Shared frailty models were used for time to event data. Overall, 52.7% of cows had no estrus event detected by an automated activity monitor system from d 7 until d 40 postpartum. Herd level prevalence of Estrus0 ranged from 37.5 to 58.4%. Estrous expression from d 7 until d 40 postpartum affected estrous duration and estrous intensity at first artificial insemination (AI). Cows in Estrus0 had the shortest duration (13.2 ± 0.33 h) compared with cows in Estrus1 (13.8 ± 0.36 h) and Estrus2 (14.8 ± 0.41 h). Cows in Estrus2 had a longer estrous duration at first postpartum AI compared with cows in Estrus1. Among Estrus0 cows, 46.2% had an estrus event with high intensity at first postpartum AI. Among cows in Estrus1 and Estrus2, 50.8 and 53.8% had an estrus event with high intensity at first postpartum AI, respectively. There was a significant difference between Estrus2 and Estrus0 and a tendency between Estrus0 and Estrus1. There was no difference between Estrus1 and Estrus2. For Estrus0, Estrus1, and Estrus2 cows, pregnancy per AI was 29.4, 30.9, and 37.8%, respectively. There was a significant difference between Estrus0 and Estrus2 and Estrus1 and Estrus2. There was no difference between Estrus0 and Estrus1. Estrous expression from d 7 until d 40 postpartum affected time to first AI and time to pregnancy. Compared with Estrus0 cows, cows in Estrus1 [hazard risk (HR) = 1.74] and Estrus2 (HR = 1.77) had an increased hazard of being inseminated within 100 DIM. There was no difference between Estrus1 and Estrus2. Median DIM to first AI were 70, 59, and 58 for cows in Estrus0, Estrus1, and Estrus2, respectively. Compared with Estrus0 cows, cows in Estrus1 (HR = 1.28) and Estrus2 (HR = 1.33) had an increased hazard of becoming pregnant within 200 DIM. There was no difference between Estrus1 and Estrus2. Median DIM to pregnancy were 127, 112, and 103 for Estrus0 cows, Estrus1 and Estrus2, respectively. In conclusion, cows with no estrous expression from 7 to 40 DIM had reduced estrous expression at first AI and inferior reproductive performance compared with cows that displayed estrous activity.