Evaluating differences in milk production, reproductive performance, and survival associated with vaginal discharge characteristics and fever in postpartum dairy cows.

The objective was to assess differences in productive and reproductive performance, and survival associated with vaginal discharge characteristics and fever in postpartum dairy cows located in Western and Southern states of the U.S.A. This retrospective cohort study included data from 3 experiments conducted in 9 dairies. Vaginal discharge was evaluated twice within 12 DIM and scored on a 5-point scale. The highest score observed for each cow was used for group assignment (VD group) as follows: VD 1 and 2 (VD 1/2; n = 1,174) = clear mucus/lochia with or without flecks of pus; VD 3 (n = 1,802) = mucopurulent with < 50% pus; VD 4 (n = 1,643) = mucopurulent with ≥50% of pus or non-fetid reddish/brownish mucous, n = 1,643; VD 5 = fetid, watery, and reddish/brownish, n = 1,800. All VD 5 cows received treatment according to each herd's protocol. Rectal temperature was assessed in a subset of VD 5 cows, and subsequently divided into Fever (rectal temperature ≥39.5°C; n = 334) and NoFever (n = 558) groups. A smaller proportion of cows with VD 5 (67.6%) resumed ovarian cyclicity compared with VD 1/2 (76.2%) and VD 4 (72.9%) cows; however, a similar proportion of VD5 and VD 3 (72.6%) cows resumed ovarian cyclicity. A smaller proportion of VD 5 (85.8%) cows received at least one artificial insemination (AI) compared with VD 1/2 (91.5%), VD 3 (91.0%), or VD 4 (91.6%) cows. Although we did not detect differences in pregnancy at first AI according to VD, fewer cows with VD 5 (64.4%) were pregnant at 300 DIM than cows with VD 1/2 (76.5%), VD 3 (76.2%), or VD 4 (74.7%). Hazard of pregnancy by 300 DIM was smaller for VD 5 compared with VD 1/2, VD 3, or VD 4 cows. A greater proportion of VD 5 cows were removed from the herd within 300 DIM compared with other VD groups. There was 760 kg lesser milk production within 300 DIM for VD 5 compared with VD 2, VD 3, and VD 4, whereas VD 2, VD 3, and VD 4 had similar milk production. We did not detect an association between fever at diagnosis of VD 5 and reproductive performance or milk production. A greater proportion of VD 5 cows without fever were removed from the herd by 300 DIM compared with VD 5 cows with fever. Differences in productive and reproductive performance, and removal of the herd were restricted to fetid, watery, and reddish/brownish vaginal discharge, which was independent of fever.

Exploring associations between the teat apex metagenome and Staphylococcus aureus intramammary infections in primiparous cows under organic directives.

The primary objective of this study was to identify associations between the prepartum teat apex microbiome and the presence of Staphylococcus aureus intramammary infections (IMI) in primiparous cows during the first 5 weeks after calving. We performed a case-control study using shotgun metagenomics of the teat apex and culture-based milk data collected longitudinally from 710 primiparous cows on five organic dairy farms. Cases had higher odds of having S. aureus metagenomic DNA on the teat apex prior to parturition compared to controls (OR = 38.9, 95% CI: 14.84-102.21). Differential abundance analysis confirmed this association, with cases having a 23.8 higher log fold change (LFC) in the abundance of S. aureus in their samples compared to controls. Of the most prevalent microorganisms in controls, those associated with a lower risk of post-calving S. aureus IMI included Microbacterium phage Min 1 (OR = 0.37, 95% CI: 0.25-0.53), Corynebacterium efficiens (OR = 0.53, 95% CI: 0.30-0.94), Kocuria polaris (OR = 0.54, 95% CI: 0.35-0.82), Micrococcus terreus (OR = 0.64, 95% CI: 0.44-0.93), and Dietzia alimentaria (OR = 0.45, 95% CI: 0.26-0.75). Genes encoding for Microcin B17 AMPs were the most prevalent on the teat apex of cases and controls (99.7% in both groups). The predicted abundance of genes encoding for Microcin B17 was also higher in cases compared to controls (LFC 0.26). IMPORTANCE: Intramammary infections (IMI) caused by Staphylococcus aureus remain an important problem for the dairy industry. The microbiome on the external skin of the teat apex may play a role in mitigating S. aureus IMI risk, in particular the production of antimicrobial peptides (AMPs) by commensal microbes. However, current studies of the teat apex microbiome utilize a 16S approach, which precludes the detection of genomic features such as genes that encode for AMPs. Therefore, further research using a shotgun metagenomic approach is needed to understand what role prepartum teat apex microbiome dynamics play in IMI risk.

Patterns of milking unit kick-off as a proxy for habituation to milking in primiparous cows.

The onset of lactation and the subsequent habituation to the milking routine is a stressful period, particularly for primiparous (PRI) cows. The objective of this study was to describe the dynamics of milking unit kick-off (KO) behavior in PRI cows during the first 3 mo of lactation, considering multiparous (MUL) cows as a reference for comparison. In addition, the potential associations between KO and milk yield and mastitis presentation were investigated. A total of 869 cows (PRI = 199; MUL = 670) on a dairy farm in northern CO were included in the analysis. Cows calving between August and November 2020 were enrolled from 3 DIM until 90 DIM. Participants were milked 3×/day in a 60-unit rotary parlor and data from each milking session were downloaded from parlor management software. Milking unit kick-off was used as a proxy for habituation to the milking procedure. Kick-off events were reported by the milking system and defined as an abrupt interruption in the milk flow during the milking process. Cow KO events were recorded for each milking session. Subsequently, occurrence of KO was analyzed by grouping the 3 consecutive milking sessions in each day and categorized as yes or no, indicating whether or not an individual kicked at least once in a given day. Data were analyzed by repeated measures ANOVA and logistic regression, including parity category, calving season, occurrence of dystocia, and their potential interactions in the models. Least squares means for daily proportions of KO were calculated considering the number of cows with KO events per day in the PRI and MUL categories. Subsequently, odds ratios for the occurrence of KO in PRI versus MUL were calculated at multiple periods of time. When KO was analyzed by DIM, proportions of KO were greater in PRI than in MUL during the whole monitoring period. In PRI, proportions of KO increased from 0.10/d to 0.20/d between 3 DIM and 15 DIM, to start decreasing around 30 DIM and remaining above MUL up to 90 DIM. On the contrary, in MUL cows, proportions of KO remained close to 0.05/d during the 90-d period. Overall, the odds of KO were greater for PRI versus MUL cows (odds ratio [95% confidence interval] = 2.07 [1.58-2.73]). No differences in milk yield were established among KO categories, while the percentage of cows affected with mastitis was greater in cows grouped in the quartile with more frequent KO events. We concluded that the relationship between days in milk and the proportion of PRI cows displaying KO was not linear, but rather KO increased during the first 2 wk postpartum before decreasing after the first month of lactation. First-parity cows had greater levels of KO than MUL cows, which is most likely associated with the process of habituation to milking during their first lactation.

Late embryo mortality in Holstein cows inseminated with Holstein or Limousine frozen semen.

The objective of this study was to compare the late embryo mortality (LEM) rate (losses approximately between 32 and 53 days of gestation) and Pregnancy Specific Protein B (PSPB) and progesterone (P4) concentrations on day 32 post AI in Holstein cows bred with either Holstein or Limousine semen. A sample size of 1082 cows per group diagnosed pregnant between 28- and 35-days post breeding was calculated. The study consisted of evaluating LEM (%) in a cohort of Holstein cows bred with Holstein semen (HO × HO) or Limousine semen (HO × LM), to compare pregnancy loss from 28 to 35 days post breeding to 50-57 days post breeding. A logistic regression model to compare embryo losses was developed considering as main explanatory variable the cohort (HO × HO embryo vs. HO × LM embryo), correcting by lactation number, breeding season, days to breeding and AI technician. HO × HO embryos had greater LEM (15.16%) than HO × LM embryos (9.79%). Cows bred in summertime had higher LEM (15.23%) than cows bred in no-summertime (9.88%). There were no differences among AI technicians. Within summertime there was no difference in LEM (%) between groups within each lactation number; yet, within no-summertime, LEM (%) was higher in HO × HO than HO × LM within each lactation number. Pregnancy SPB optical densities were significantly greater in the HO × HO than in the HO × LM (p = .023) group; yet, the concentration of P4 was not different between groups (p > .05).

Combined effect of purulent vaginal discharge and anovulation on pregnancy status in a large multi-state population of Holstein cows.

The objective of this observational prospective cohort study was to evaluate the combined effect of purulent vaginal discharge (PVD) and anovulation (ANOV) on the reproductive performance of a large multi-state population of Holstein cows. Data were prospectively collected from 11,729 cows in 16 herds located in 4 regions in the United States [Northeast (4 herds), Midwest (6), Southeast (1), and Southwest (5)]. Cows were enrolled at calving and monitored weekly for disease occurrence, reproductive events, and survival. Prevalence of PVD was evaluated at 28 ± 3 d in milk and defined by the presence of mucopurulent to fetid vaginal discharge. Resumption of ovarian cyclicity was determined via transrectal ultrasonography at 40 ± 3 and 54 ± 3 d postpartum. Pregnancy diagnosis was performed by ultrasonography on d 32 ± 3 after artificial insemination (AI) and reconfirmed at d 60 ± 3 of gestation. Pregnancy loss (PL) was defined as a cow diagnosed pregnant at 32 ± 3 but nonpregnant at 60 ± 3 d after AI. The association of PVD and ANOV with pregnancy traits was analyzed using 4 PVD-cyclicity categories that considered the following combinations: NPVD-CYC = absence of PVD and cycling; PVD-CYC = presence of PVD and cycling; NPVD-ANOV = absence of PVD and anovular; and PVD-ANOV = presence of PVD and anovular. Multiple logistic regression and Cox proportional regression were used for the analysis of potential associations between PVD and cyclicity categories and pregnancy at first AI (PAI1), days from calving to pregnancy, and PL at first AI. The odds (95% confidence intervals) of pregnancy increased from cows in the PVD-ANOV category (reference category) to cows in NPVD-ANOV [2.09 (1.62-2.50)], PVD-CYC [2.52 (2.02-3.14)], and NPVD-CYC [3.46 (2.84-4.23)]. Similarly, days from calving to pregnancy were less for NPVD-CYC, followed by PVD-CYC, NPVD-ANOV, and PVD-ANOV (121.4, 137.2, 137.3, and 157.4 d, respectively). On the contrary, no clear association was identified between groups and PL. The results indicate that both PVD and ANOV had a negative impact on PAI1 and days from calving to pregnancy. The results indicated a variable magnitude in the negative impact on the reproductive traits analyzed when both conditions were combined.

Waiting time in the premilking holding pen and subsequent lying and walking behaviors of Holstein cows.

The objective of this study was to assess the effect of waiting time (WT) in the premilking holding pen on the subsequent lying and walking behaviors of Holstein cows. A total of 108 cows (multiparous n = 95; primiparous n = 13), milked 3 times/d [AM (0700 h), PM (1500 h), and nighttime (2300 h)] were monitored for individual WT, which was defined as the time elapsed between the entrance of the first cow to the milking stall in a rotary milking system and the entrance of each subsequent cow housed in the same milking pen. Data for lying time (min), standing time (min), lying bouts (no.), and walking behavior (no. of steps) in 15-min intervals were collected using a commercial 3-dimensional accelerometer affixed on the hind left leg of each cow from early to mid lactation. Mean values (min/h, no./h) for each behavior calculated between consecutive milkings were compared among 3 WT categories: cows with WT ≤30 min (WT1), WT 30 to 60 min (WT2), and WT >60 min (WT3). The overall mean (SD) of WT during the study period was 31.2 (23.4) min, and the distribution of WT1, WT2, and WT3 in all cows through the monitoring period was 50.9, 43.3, and 5.7%, respectively. No significant effects were established for overall WT on any of the behaviors under analysis. However, subsequent analysis by milking shift indicated that, during the period following the night milking, cows in WT1 exhibited the shortest lying time, whereas cows in WT3 had the greatest number of lying bouts. No effect of WT category was observed on the number of steps after any of the milking shifts. Results of this study indicate a moderate effect of WT on lying behavior after the nighttime milking. Variables such as temperature-humidity index, parity, and time of day affected cow behavior and should be considered when evaluating the impact of routine management practices, such as milking, on commercial farms.

Effect of a 2-step probiotic program on digestive health and performance of Holstein heifer calves.

Managing the composition of the bacterial communities in the digestive tract with the use of probiotics that enhance protective microflora could result in positive effects on health and performance of calves. The objective of this study was to evaluate a 2-step probiotic program (added to colostrum and milk) on the digestive health and growth of preweaning Holstein heifers. A randomized clinical trial was conducted from July to October 2020 in a calf rearing facility in Colorado. Calves were housed in pairs sharing the same treatment in 2 polyethylene hutches within a common area of 4.50 m2. A total of 232 calves were enrolled at birth and randomly allocated into 2 treatment groups [control (CTR) = 116; treatment (PB) = 116] and followed until weaning (64 ± 3 d). Treatment consisted of 2 formulations of a multistrain bacterial-based probiotic added in colostrum (PBF1) and milk (PBF2). Treatment calves received 2 g of PBF1 added to each colostrum feeding, and 1 g of PBF2 added to the milk at the morning feeding 3 times per week up to weaning. Calf weight was collected at birth, at 30 d of age, and at weaning. Serum total protein was determined at age 3 ± 1 d and a health assessment was completed 3 times per week. Statistical analyses were performed using SAS, with calf considered the experimental unit clustered by housing pair. Cox proportional hazard analysis and time to event analysis were used to compare time to the first diarrhea event and time to recovery between treatment groups. The total number of diarrhea events and culling were assessed by Poisson regression and logistic regression analysis, respectively. Linear regression was performed to evaluate differences in average daily gain by treatment group. Overall, the mean (± standard error) temperature humidity index (THI) was 75 (± 0.44) units during the study period, which resulted in continuous exposure to heat stress in the day hours. No difference between treatment groups was observed in serum total protein. The overall incidence of diarrhea was 96.6% (CTR = 99%, PB = 95%); no differences were determined in the hazard of a first diarrhea episode or in the median time to the first diarrhea event (11 d in both groups). Similarly, no differences were found in the likelihood of recovery or in the median time to recovery from diarrhea (7 d in both groups). Likewise, treatment group was not associated with the number of diarrhea events, mean (± standard error, g/d) average daily gain from birth to weaning [CTR = 562.5 (13.9); PB = 570.8 (13.5)], or with the odds of culling. The most prevalent pathogen isolated from feces was Clostridium, which tended to be more frequent in CTR calves than PB calves. In conclusion, the probiotic program did not affect the incidence of enteric disease or the growth of preweaning heifer calves.

Dynamics of automatically generated body condition scores during early lactation and pregnancy at first artificial insemination of Holstein cows.

The objective of this study was to characterize the association between body condition score (BCS) and BCS change (ΔBCS), determined by an automated camera system at multiple time points, and the subsequent pregnancy per first artificial insemination (P/AI1) of Holstein cows. A retrospective observational study was completed using data collected from 11,393 lactations in 7,928 Holstein cows calving between April 2019 and March 2021 in a commercial dairy operation located in Colorado. Cows were classified as primiparous or multiparous. Scores generated by BCS cameras at dry-off, calving, 21 days in milk (DIM), 56 DIM, and first artificial insemination were selected for the analyses and subsequently categorized as low (≤lower quartile), moderate (interquartile range), and high (≥upper quartile). Changes in BCS were calculated by periods of interest as change from dry-off to calving (multiparous cows); change from calving to 21 DIM; change from calving to 56 DIM; and change from calving to first artificial insemination and assigned into categories as large loss of BCS (top 25% of cows losing BCS); moderate loss (bottom 75% of cows losing BCS); no change (ΔBCS = 0); or gain of BCS (ΔBCS > 0). Data were examined in primiparous and multiparous cows separately using logistic regression and time-to-event analyses. Initial univariable models were followed by multivariable models that considered calving season, occurrence of disease, and milk yield up to 60 DIM as covariables. The logistic regression analyses indicated that in both parity groups the associations between BCS category and P/AI1 were more evident at 21 DIM, 56 DIM, and first artificial insemination, with lower odds of P/AI1 in cows in the low BCS category. Likewise, cows with large loss in BCS between calving and 21 DIM, calving and 56 DIM, and calving and first artificial insemination had lower odds of P/AI1 compared with other categories of ΔBCS within the same period of interest. Similarly, survival analyses evidenced that cows in the low BCS category required more time to get pregnant. In agreement, differences in the dynamics of the average daily BCS during the first 90 DIM were evident when cows were grouped by first AI outcome (pregnant vs. nonpregnant) and by their time to pregnancy category (<90 DIM; 91-150 DIM; or >150 DIM), with cows with reduced fertility showing lower BCS up to 90 DIM. Overall, low BCS and more pronounced reductions in BCS occurring closer to first artificial insemination resulted in lower odds of pregnancy per artificial insemination.

Effect of fish oil and canola oil supplementation on immunological parameters, feed intake, and growth of Holstein calves.

Supplemental n-3 fatty acids (FA) may support better immune responses than n-6 and n-9 FA in dairy calves. The objective was to evaluate the effect of n-3 FA, supplemented as a fish oil product (FO) in the milk replacer (MR), in comparison to n-6 and n-9 FA, supplemented as canola oil (CO), on body weight (BW), daily gain, and immunological parameters of preweaning Holstein calves. The study was conducted from September to December 2019. Calves were randomly allocated to a control group (n = 15; BW = 36.2 ± 1.5 kg; mean ± SEM) supplemented daily with 30 mL of CO and to an experimental group (n = 15; BW = 36.3 ± 1.5 kg) supplemented with 60 g of a product containing 30 g of FO. Both treatments were added to the MR during the morning feeding. All calves were fed 4 L of MR at 12.5% solids at 0700 and 1600 h for wk 1, 6 L from wk 2 to 7, and 3 L once daily (0700 h) during wk 8 until weaning (56 d). Blood samples were collected at 7, 14, 21, 28, 35, 42, 49, and 56 d of age for serum haptoglobin, TNF-α, IL-1ß, and protectin. Dry matter intake was recorded in all experimental calves daily. Seroneutralization titers to vaccination against viral diseases (infectious bovine rhinotracheitis, parainfluenza 3, bovine viral diarrhea, and bovine respiratory syncytial virus) were determined. Mixed models for repeated measures were developed to analyze variables over time. Seroneutralization titers were analyzed by the Kruskal-Wallis test. The other variables were compared by a generalized linear model. Serum FA profile at 35 d of age showed that FO supported higher concentrations of n-3 FA than CO. Final BW [65.2 vs. 62.0 kg, standard error of the mean (SEM) = 2.1 kg] and average daily gain (0.52 vs. 0.46 kg/d, SEM = 0.1 kg/d) tended to be higher for the FO than the CO group. An interaction of treatment × day for dry matter intake was observed, especially during weaning (2.17 kg vs. 1.94 kg, SEM = 0.158 kg, for FO and CO group, respectively). Blood lactate (mmol/L) was higher in the CO than in the FO group at d 7. Haptoglobin and IL-1ß were higher for the CO group on d 14 than the FO group. The TNF- α concentrations for the FO group were reduced over time, whereas the concentrations in the CO group remained constant. Protectin was higher in the FO group on d 14, but was lower on d 28, 35, and 49. Seroneutralization antibody titers postvaccination for the PI3 virus were higher for the FO than the CO group. In conclusion, calves supplemented with FO had lower concentrations of blood lactate, haptoglobin, IL-1ß and TNF-α than calves supplemented with CO during the study period. The FO supplementation had a higher DMI than CO supplementation. Results of this trial should be interpreted with caution due to the lack of a negative control group as well as the lower birth weight and growth rate observed under heat stress conditions.

Effect of body condition change and health status during early lactation on performance and survival of Holstein cows.

Body condition score (BCS) and disease records are commonly available in dairy operations. However, the effect of BCS changes (ΔBCS) considering specific health profiles has not been investigated extensively. The objective of this study was to assess the effects of different levels of ΔBCS on fertility, milk yield, and survival of Holstein cows diagnosed with reproductive disorders (REP; dystocia, twins, retained fetal membranes, metritis, and clinical endometritis), other health disorders (OTH; subclinical ketosis, left displaced abomasum, lameness, clinical mastitis, and respiratory disease), or with no disease events (HLT) within 40 days in milk (DIM). Data included lactation information from 11,733 cows calving between November 2012 and October 2014 in 16 herds across 4 geographical regions in the United States (Northeast, Midwest, Southwest, Southeast). Cows were evaluated for BCS at 5 ± 3 DIM (BCS5) and at 40 ± 3 DIM (BCS40) and the difference between BCS40 and BCS5 was classified as excessive loss of BCS (EL; ΔBCS ≤-0.75), moderate loss (ML; ΔBCS = -0.5 to -0.25), no change (NC; ΔBCS = 0), or gain of BCS (GN; ΔBCS ≥0.25). Multivariable logistic regression was used for assessing potential associations between the outcomes of interest and ΔBCS and health. The effect of the interaction term ΔBCS by health group was not statistically significant for any of the study outcomes. The odds of resumption of ovarian cyclicity (ROC), in GN, NC, and ML cows were 1.94 (95% CI: 1.57-2.40), 1.59 (1.28-1.97), and 1.27 (1.10-1.47) times greater than the odds of ROC in EL cows, respectively. The odds of pregnancy at 150 DIM (P150) in GN cows were 1.61 (1.20-2.17) times greater than the odds of P150 in EL cows. Cows with REP or OTH disorders had smaller odds of ROC compared with HLT cows [REP: OR = 0.65 (0.56-0.76) and OTH: OR = 0.79 (0.68-0.92)]. For pregnancy outcomes, REP cows had smaller odds of pregnancy at the first artificial insemination compared with HLT cows [0.70 (0.58-0.84)]. Similarly, REP cows had smaller odds of being diagnosed pregnant by 150 and 305 DIM compared with HLT cows [P150: 0.73 (0.59-0.87), P305: 0.58 (0.49-0.69)]. Overall, average daily milk within the first 90 DIM was greater in EL (39.5 ± 1.13 kg/d) and ML (38.9 ± 1.11 kg/d) cows than in NC (37.8 ± 1.12 kg/d) and GN (36.2 ± 1.12 kg/d) cows. On the other hand, average daily milk within the first 90 DIM was lower in REP (37.0 ± 1.11 kg/d) cows compared with OTH (38.7 ± 1.12 kg/d) and HLT cows (38.6 ± 1.11 kg/d). The magnitude of ΔBCS and the health status of early lactation cows should be considered when assessing subsequent cow performance and survival.

The economic cost of metritis in dairy herds.

The objective of this study was to estimate the cost of metritis in dairy herds. Data from 11,733 dairy cows from 16 different farms located in 4 different regions of the United States were compiled for up to 305 d in milk, and 11,581 cows (2,907 with and 8,674 without metritis) were used for this study. Metritis was defined as fetid, watery, red-brownish vaginal discharge that occurs ≤21 d in milk. Continuous outcomes such as 305-d milk production, milk sales ($/cow), cow sales ($/cow), metritis treatment costs ($/cow), replacement costs ($/cow), reproduction costs ($/cow), feeding costs ($/cow), and gross profit per cow ($/cow) were analyzed using mixed effect models using the MIXED procedure of SAS (SAS Institute Inc., Cary, NC). Gross profit was also compared using the Kruskal-Wallis test. Dichotomous outcomes such as pregnant and culling by 305 d in milk were analyzed using the GLIMMIX procedure of SAS. Time to pregnancy and culling were analyzed using the PHREG procedure of SAS. Models included the fixed effects of metritis, parity, and the interaction between metritis and parity, and farm as the random effect. Variables were considered significant when P ≤ 0.05. Metritis cost was calculated by subtracting the gross profit of cows with metritis from the gross profit of cows without metritis. A stochastic analysis was performed with 10,000 iterations using the observed results from each group. Milk yield and proportion of cows pregnant were lesser for cows with metritis than for cows without metritis, whereas the proportion of cows leaving the herd was greater for cows with metritis than for cows without metritis. Milk sales, feeding costs, residual cow value, and gross profit were lesser for cows with metritis than for cows without metritis. Cow sales and replacement costs were greater for cows with metritis than for cows without metritis. The mean cost of metritis from the study herds was $511 and the median was $398. The stochastic analysis showed that the mean cost of a case of metritis was $513, with 95% of the scenarios ranging from $240 to $884, and that milk price, treatment cost, replacement cost, and feed cost explained 59%, 19%, 12%, and 7%, respectively, of the total variation in cash flow differences. In conclusion, metritis caused large economic losses to dairy herds by decreasing milk production, reproduction, and survival in the herd.

Lying and stepping behaviors around corrective or therapeutic claw trimming.

The objectives of this study were to test differences in lying and stepping behaviors in lactating cows with altered gait or other signs of pain in the day preceding either corrective claw trimming (CCT) or therapeutic claw trimming (TCT; d -1), including healthy controls (HC) as a reference group, and to analyze the associations between categories of lying and stepping behaviors at d -1 and subsequent submission to CCT or TCT (d 0). In addition, we characterized variations in lying and stepping behaviors within 7 d relative to CCT or treatment for a specific foot condition (TCT). A total of 310 Holstein cows in a USDA-certified organic dairy herd in northern Colorado were enrolled. Cows were affixed with an accelerometer (IceQube, IceRobotics) below the fetlock of one hind leg at 12 ± 8 d in milk and monitored for 6 mo for daily activity [lying time (LY; min/d), lying bouts (LB; no./d), and steps (ST; no./d)]. Cows with altered gait or other signs of pain were submitted to claw trimming (CT; d 0) and differentiated as receiving only corrective interventions (CCT) or as being treated for a lameness disorder (TCT). Concurrent activity from cows not submitted to CT was considered a reference from HC. Daily averages for each activity within ±7 d relative to CT were compared among the 3 groups using linear mixed models (PROC MIXED of SAS; SAS Institute Inc.). Logistic regression analyses with a logit link function (PROC GLIMMIX of SAS) were used to test the associations between categories (lower quartile or less, interquartile range, and higher quartile or greater) of lying and stepping behaviors (LY, LB, and ST) preceding CT (d -1) and subsequent submission to CCT or TCT (d 0). On d -1, average (± standard error) LY was greater in TCT (631 ± 34 min/d) than in CCT (568 ± 22 min/d) and HC (581 ± 13 min/d). Overall, LB were not statistically different among groups (CCT = 18.7 ± 0.02 bouts/d; TCT = 19.6 ± 1.17 bouts/d; and HC = 19.1 ± 0.43 bouts/d). Daily ST were lower in TCT (1,810 ± 126 steps/d) than in CCT (2,803 ± 63 steps/d) and HC (2,542 ± 103 steps/d). The odds (confidence interval in parentheses) of TCT were greater in cows in the high LY [3.27 (2.47-4.33)] and high LB [2.31 (1.77-3.02)] categories at d -1. The low ST category was associated with increased odds of CCT [3.32 (1.96-5.65)] and TCT [8.48 (6.49-11.1)]. We concluded that, under the current experimental conditions, cows requiring TCT evidenced greater overall LY and lower daily ST the day before CT. However, these differences were not consistent throughout days in milk categories within the lactation. Type of CT was partially associated with category of activity the day preceding CT, and the associations were more evident for TCT. The magnitude of the behavioral changes before and after CT and the recovery times posttreatment to approach HC behavioral levels varied depending on specific conditions.

A randomized controlled clinical trial on the effect of acupuncture therapy in dairy cows affected by pyometra.

Pyometra (PYO) is a reproductive disease characterized by the accumulation of purulent or mucopurulent material within the uterine lumen, in the presence of an active corpus luteum (CL). As the CL continues secreting progesterone, PYO would develop following endometritis. Due to prohibited use of artificial luteolytic hormones in US certified organic dairies, conventional therapies consisting of administration of prostaglandin F2α, are not applicable. The objective of this study was to evaluate the efficacy of two acupuncture procedures on the treatment of persistent CL in cows with PYO. We hypothesized that acupuncture would reduce CL diameter and serum progesterone (P4) concentrations, leading to regression of PYO. Holstein cows with PYO, at an USDA certified organic dairy farm in Northern Colorado, were enrolled in a randomized controlled clinical trial and assigned to 1 of 3 treatments: (1) control pyometra (CP; no treatment; n = 17); (2) electroacupuncture (EAP; n = 15); and (3) laser acupuncture (LAP; n = 15). Each cow received three 9-min (EAP) or 20-min (LAP) acupuncture sessions in alternate days. All study cows had blood samples collected for determination of serum progesterone concentration at d0, and at d2, d4, d11, d18, and d25 after first treatment. The ovaries were scanned by transrectal ultrasonography at -3d, d0, d2, d4, d11, and d18 to determine the diameter of the CL. The effect of treatment in the outcome variables was evaluated by ANOVA and by repeated measures analyses, accounting for baseline data (CL diameter and progesterone serum concentration). Average ± SE change in CL diameter from d0 to d18 were 0.94 ± 1.0 mm, 0 ± 1.0 mm, and - 0.33 ± 1.0 mm for CP, EAP, and LAP, respectively. The repeated measures analysis indicated no significant differences for CL diameter among groups. None of the study cows had serum progesterone values <1 ng/ml by the end of the monitoring period (d25) and average ± SE change from d0 to d25 were - 4.0 ± 1.97 ng/ml, -0.76 ± 2.1 ng/ml, and 3.24 ± 1.9 ng/ml for CP, EAP, and LAP, respectively. The repeated measures analysis indicated no significant differences for serum progesterone concentrations among groups. On farm cow records reviewed 150 d after treatment indicated that 2 cows in EAP and 1 cow in LAP conceived 38 d, 68 d, and 38 d, after treatment completion. In conclusion, acupuncture was not an effective treatment for persistent CL in cows with PYO, during the monitoring period.

Early-lactation diseases and fertility in 2 seasons of calving across US dairy herds.

The objective of this study was to characterize incidences of health disorders during early lactation in a large population of Holstein cows calving in 2 seasons across multiple US dairy herds. In addition, cumulative effects of combinations of health-related events on fertility and survival by season of calving and parity number were tested. Data were prospectively collected from a total of 11,729 cows in 16 herds located in 2 regions in the United States [north (7,820 cows in 10 herds) and south (3,909 cows in 6 herds)]. Cows were enrolled at parturition and monitored weekly for disease occurrence, reproductive events, and survival. Health-related events were grouped into reproductive disorders (REP; dystocia, twins, retained fetal membranes, metritis, and clinical endometritis) and other disorders (OTH; subclinical ketosis, mastitis, displaced abomasum, and pneumonia). Counts of health events within 50 d postpartum were added into each of the groups and categorized as 0, 1, 2, 3, and ≥4 for REP and 0, 1, 2, and ≥3 for OTH. Multivariable logistic regression was used for testing potential associations between categories of disease occurrence and outcome variables, including resumption of ovarian cyclicity, pregnancy per artificial insemination (AI), pregnancy loss, and survival up to and after 50 DIM. The incidence of disease varied with season of calving and parity, and these 2 variables were associated with the reproductive and survival outcomes. The size of the detrimental effect of disease incidence on reproduction and survival depended on disease group and varied for each specific outcome. Resumption of ovarian cyclicity decreased as incidences of disorders increased in both REP and OTH categories. Pregnancy at first AI also was smaller in greater number of REP categories, but the effect of number of OTH categories on pregnancy at first AI was not consistent. Similarly, pregnancy loss at first AI was not affected consistently by REP or OTH. Survival was reduced by REP and OTH. The magnitude of these negative effects was variable, depending on season of calving and parity, but consistently increased with the number of health events during early lactation.

Incidence and risk factors for reproductive disorders in organic certified dairies.

The objective was to determine the lactational incidence risks of reproductive disorders (RD) including dystocia (DYS), retained fetal membranes (RFM), puerperal metritis (PM), clinical endometritis (CE), pyometra (PYO), late pregnancy loss (LPL), and culling due to reproductive reasons (CRR) in large organic certified dairies with predominant use of Holstein genetics. In addition, potential risk factors for RD, including parity category, milk yield in the previous lactation, season of dry-off, access to grazing at dry-off, length of the dry period (DPL), length of the close-up period (CUPL), calculated gestation length (CGL), season of calving, access to grazing at calving, concurrent health conditions, and total number of artificial inseminations were analyzed. A retrospective study was performed using information from 8 organic certified herds located in northern Colorado and west Texas, ranging from 900 to 6,000 milking cows. Cow information consisted of finalized lactations starting with calvings occurring between January 1, 2016, and December 31, 2018. Data were analyzed using time-to-event analyses and multivariable logistic regression. Clinical endometritis had the greatest lactational incidence risk (incidence risk = 10.1%; 95% CI = 9.8-10.3%), followed by DYS (8.8%; 8.5-9.1%), PM (4.9%; 4.7-5.1%), PYO (4.0%; 3.8-4.1%), LPL (3.6%; 3.4-3.7%), and RFM (2.0%; 1.8-2.1%). The incidence risk of CRR was 7.5% (7.3-8.7%), representing 29.1% (28.4-30.0%) of all culling reasons. Median time from calving to diagnosis of PM, CE, PYO, and LPL were 6, 23, 111, and 228 d, respectively. Median time for CRR was 391 d. The main risk factors for RD were DPL, CGL, and CUPL. Season of dry-off was a risk factor for RFM, PM, CE, LPL, and PYO. Short DPL (<30 d) was associated with increased odds (95% CI) of RFM [2.1 (1.6-2.7)], PM [1.4 (1.2-1.7)], and LPL [2.5 (1.5-4.2)]. Shortened CGL (<270 d) increased the odds of DYS [1.5 (1.3-1.8)], RFM [3.2 (2.4-4.1)], and PM [2.4 (1.7-3.3)], whereas extended CGL (>285 d) increased the odds of DYS [1.5 (1.4-1.7)], PM [1.5 (1.2-2.0)], CE [1.5 (1.1-2.0)], and CRR [1.2 (1.10-1.4)]. Shortened CUPL was associated with increased odds of all the RD. Cows spending less than 7 d in the close-up groups had greater odds of DYS [1.4 (1.1-1.8)], RFM [2.1 (1.8-2.4)], PM [1.9 (1.6-2.1)], PYO [1.5 (1.2-1.8)], LPL [2 (1.7-2.3)], and CRR [1.4 (1.04-1.8)] compared with cows spending more than 14 d in the close-up group. Season of calving was associated with the odds of PM [summer = 1.13 (1.01-1.2)], PYO [summer = 1.7 (1.1-2.5)], and LPL ([fall = 0.8 (0.6-0.96)] compared with winter calving. Other risk factors affecting RD were parity number, total number of artificial inseminations, and concurrent health conditions. Overall, lactational incidence risks of reproductive diseases were comparable with those reported in conventional herds. The identified risk factors for RD open opportunities for interventions, especially focused in precalving management, estimation of due dates, and closer monitoring of pregnancies from natural services in large organic certified dairy herds.

Associations of reproductive indices with fertility outcomes, milk yield, and survival in Holstein cows.

The study is part of a research effort investigating potential associations between genomic variation and fertility of Holstein cows. The objective was to compare the reproductive performance of Holstein cows in 3 categories of 2 reproductive indices (RI) that were developed for the allocation of cows in a ranking for potential fertility, based on the predicted probability of pregnancy. The associations between categories of the developed indices and multiple fertility variables in a large multistate population of Holstein cows were tested. In addition, we analyzed associations among the RI categories with milk yield and survival. Based on phenotypic information from individual cows, 2 reproductive indices (RI1 and RI2) were developed, representing a predicted probability that a cow will become pregnant at first artificial insemination postpartum, as a function of explanatory variables used in a logistic model. Data from a total of 11,733 cows calving in 16 farms located in 4 regions of the United States (Northeast, Midwest, Southeast, and Southwest) were available. Cows were enrolled at parturition and monitored weekly for reproductive events, health status, milk yield, and survival. To develop the indices, potential significant effects were initially tested by univariate analyses. Effects with P ≤ 0.05 were offered to the multivariate analysis, and the final models were determined through backward elimination, considering potentially significant interactions. The final model for RI1 included the random effect of farm and a complement of significant fixed effects as explanatory variables influencing a pregnancy outcome: (1) incidence of retained fetal membranes; (2) metritis; (3) clinical endometritis; (4) lameness at 35 days in milk (DIM); (5) resumption of postpartum ovulation by 50 DIM; (6) season of calving; and (7) parity number. The model for RI2 included (1) parity number; (2) body condition score at 40 DIM; (3) incidence of retained fetal membranes; (4) metritis; (5) resumption of postpartum ovulation by 50 DIM; (6) region; (7) subclinical ketosis; (8) mastitis; (9) clinical endometritis; and (10) milk yield at the first milk test after calving; as well as the interaction effects of postpartum resumption of ovulation by 50 DIM × region; mastitis × region; and milk yield at the first milk test after calving × parity number. Multivariate logistic regression, ANOVA, and survival analysis were used to test the correspondence between the resulting RI and individual fertility, milk yield, and survival from the population. To facilitate the analyses, the resulting RI values were categorized as low for cows in the lowest quartile, medium for cows within the interquartile range, or high for cows in the top quartile. We found consistent agreement between categories of the predicted RI and the measures of fertility and survival collected from individual cows. We conclude that the proposed RI represent a viable approach to refine the allocation of cows into potential low- and high-fertility populations.

Efficacy of non-antibiotic treatment options for digital dermatitis on an organic dairy farm.

The objective of this study was to evaluate the efficacy of two non-antibiotic treatment options for digital dermatitis (DD) on an organic certified dairy farm. A randomized clinical trial was conducted using 70 multiparous Holstein cows with an early DD lesion at a USDA certified organic dairy farm in Northern Colorado, USA. Cows were enrolled in the study based on the presence of early DD lesions (scores M1 and M2) and randomly assigned to one of three treatments: (1) topical application of copper sulfate and iodine (CUI); (2) topical application of honey and iodine (HOI); and (3) control subject to no treatment (CON). Cows were evaluated at enrolment and on days 3, 12, 28, and 120 post treatment for pain and lesion size and received a locomotion and a lesion score. Cure was defined as the transition from active to non-active stages (M1/M2 to M0 or M4). The formulations had variable effects on the treatment of DD. The cure rate was numerically higher for CUI on all follow up days. The proportion of cows experiencing pain on d3 after treatment was greater in CON, followed by HOI and CUI. However, this proportion increased in HOI during the follow up period. The CUI group had a greater reduction in lesion size and larger lesions persisted in HOI. Non-antibiotic treatment formulations were partially effective in the treatment of DD in organic dairy cows: The two non-antibiotic formulations resulted in an earlier transition to mature lesions compared with the control group. The CUI combination was the most effective treatment in reducing lesion size, pain, and lameness in affected cows. However, this combination had short-term efficacy, which did not persist throughout the duration of the study. The HOI combination produced only transient reduction in lesion size.

Genome-enable prediction for health traits using high-density SNP panel in US Holstein cattle.

The objective of this study was to compare accuracies of different Bayesian regression models in predicting molecular breeding values for health traits in Holstein cattle. The dataset was composed of 2505 records reporting the occurrence of retained fetal membranes (RFM), metritis (MET), mastitis (MAST), displaced abomasum (DA), lameness (LS), clinical endometritis (CE), respiratory disease (RD), dystocia (DYST) and subclinical ketosis (SCK) in Holstein cows, collected between 2012 and 2014 in 16 dairies located across the US. Cows were genotyped with the Illumina BovineHD (HD, 777K). The quality controls for SNP genotypes were HWE P-value of at least 1 × 10-10 ; MAF greater than 0.01 and call rate greater than 0.95. The FImpute program was used for imputation of missing SNP markers. The effect of each SNP was estimated using the Bayesian Ridge Regression (BRR), Bayes A, Bayes B and Bayes Cπ methods. The prediction quality was assessed by the area under the curve, the prediction mean square error and the correlation between genomic breeding value and the observed phenotype, using a leave-one-out cross-validation technique that avoids iterative cross-validation. The highest accuracies of predictions achieved were: RFM [Bayes B (0.34)], MET [BRR (0.36)], MAST [Bayes B (0.55), DA [Bayes Cπ (0.26)], LS [Bayes A (0.12)], CE [Bayes A (0.32)], RD [Bayes Cπ (0.23)], DYST [Bayes A (0.35)] and SCK [Bayes Cπ (0.38)] models. Except for DA, LS and RD, the predictive abilities were similar between the methods. A strong relationship between the predictive ability and the heritability of the trait was observed, where traits with higher heritability achieved higher accuracy and lower bias when compared with those with low heritability. Overall, it has been shown that a high-density SNP panel can be used successfully to predict genomic breeding values of health traits in Holstein cattle and that the model of choice will depend mostly on the genetic architecture of the trait.

Use of milk electrical conductivity for the differentiation of mastitis causing pathogens in Holstein cows.

Mastitis is one of the most prevalent and costly diseases in dairy cattle. Key components for adequate mastitis control are the detection of early stages of infection, as well as the selection of appropriate management interventions and therapies based on the causal pathogens associated with the infection. The objective was to characterize the pattern of electrical conductivity (EC) in milk during intramammary infection, considering specific mastitis-causing pathogen groups involvement. Cows (n = 200) identified by an in-line mastitis detection system with a positive deviation ≥15% in the manufacturer's proprietary algorithm for EC (high electrical conductivity (HEC)) were considered cases and enrolled in the study at the subsequent milking. One control (CON) cow, within normal ranges for EC, was matched to each case. A composite milk sample was collected aseptically from each cow for bacteriological culture. Milk yield (MY) and EC were recorded for each milking during ±7 days relative to enrollment. Milk cultures were categorized into gram positive (GP), gram negative (GN), other (OTH) and no growth (NOG). Data were submitted for repeated-measures analysis with EC as the dependent variable and EC status at day -1, bacteriological culture category, parity number, stage of lactation and days relative to sampling as main independent variables. Average (± standard error (SE)) EC was greater in HEC than in CON cows (12.5 ± 0.5 v. 10.8 ± 0.5 mS/cm) on the day of identification (day -1). Milk yield on day -1 was greater in CON than in HEC (37.6 ± 5.1 v. 33.5 ± 5.2 kg). For practical management purposes, average EC on day -1 was similar for the different bacteriological culture categories: 11.4 ± 0.6, 11.7 ± 0.5, 12.3 ± 0.8 and 11.7 ± 0.5 mS/cm in GN, GP, OTH and NOG, respectively. Parity number was only associated with day -1 EC in HEC group, with the greatest EC values in parity 3 (12.3 ± 0.3 mS/cm), followed by parity 2 (11.9 ± 0.2 mS/cm), parity >3 (11.6 ± 0.5 mS/cm) and primiparous cows (11.2 ± 0.2 mS/cm). An effect on EC for the interaction of day relative to identification by pathogen gram category was observed. The same interaction effect was observed on daily MY. Overall, the level of variation for MY and EC between- and within-cows was substantial, and as indicated by the model diagnostic procedures, the magnitude of the variance in the cows in the CON group resulted in deviations from normality in the residuals. We concluded that characteristic temporal patterns in EC and MY in particular pathogen groups may provide indications for differentiation of groups of mastitis-causing pathogens. Further research to build detection models including EC, MY and cow-level factors is required for accurate differentiation.