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.

Milk component ratios and their associations with energy balance indicators and serum calcium concentration in early-lactation spring-calving pasture-based dairy cows.

Indirect assessment of metabolic status using milk samples provides a non-invasive and objective tool for cow-level health monitoring. Milk fat-to-protein ratio (FPR) has been commonly evaluated as an indirect measure for negative energy balance (EB) in confined dairy cows. However, milk component ratios have not been explored for their association with pasture-based cows' metabolic status. The objectives of this observational study were to 1) describe milk component ratios from 0 to 45 d postpartum, 2) evaluate the associations between milk component ratios [FPR, fat-to-lactose (FLR), protein-to-lactose (PLR)] and indicators of EB (serum ß-hydroxybutyrate (BHB) concentration at 5-45 d postpartum and body condition score (BCS) change during the transition period), and 3) evaluate the associations between milk component ratios and serum Ca concentration 0-4 d postpartum in spring-calving dairy cows from pasture-based commercial farms. Milk component ratios were determined on samples collected before AM or PM milkings from 548 cows at 0-45 d postpartum (n = 970). Serum BHB and Ca determinations were performed in blood samples collected at the time of milk sample collection at 5-45 d postpartum (n = 918) and 0-4 d postpartum (n = 50), respectively; and BCS change was calculated using BCS assigned between 29 d prepartum and 45 d postpartum (n = 851). Cows' calving date, parity (1st, 2nd-3rd or ≥ 4th) and breed (Holstein-Friesian or dairy crossbred) information was obtained from the farm records. Data was analyzed by multiple linear regression. Average milk FPR, FLR and PLR were 0.70, 0.53 and 0.72, respectively. Milk FPR linearly increased while milk FLR linearly decreased postpartum both at a rate of 0.004 units per day; milk PLR decreased 0.05 units per day for the first 30 d postpartum and moderately increased afterward. Milk FPR and FLR were 0.71 and 0.52 units lower before AM than PM milking, respectively; while milk PLR was similar before AM and PM milking. Milk FPR and FLR were 0.07 to 0.10 units higher for 2nd-3rd compared with 1st and ≥ 4th parity cows. Milk PLR was 0.03 units greater for ≥ 4th compared with 2nd-3rd and 1st parity cows. Further, crossbred cows had 0.07, 0.08 and 0.03 higher milk FPR, FLR and PLR than Holstein-Friesian cows, respectively. Moderate to high P-values along with moderate to small estimated slopes and wide 95% confidence intervals were observed for the associations between milk component ratios and indicators of EB. A positive linear association was observed between milk FPR and serum Ca concentration within 4 d postpartum; milk FPR increased 0.31 units per each mmol/L increase in serum Ca concentration. Cows with low serum Ca concentration within 4 d postpartum had 0.27 units lower milk FPR compared with cows at or above the threshold (2.12 mmol/L), and tended to have 0.15 units lower milk FPR compared with cows at or above the threshold (2.00 mmol/L). In conclusion, further research is needed to reach conclusions on the association between milk component ratios determined before milking and EB indicators. The potential of milk FPR for monitoring blood Ca status warrants further investigation in early-lactation pasture-based dairy cows.

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 United States. 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 vaginal discharge score observed for each cow was used to allocate them into 1 of 5 possible groups (VD group) as follows: VD 1 and 2 (VD 1/2; n = 1,174) = clear mucus or 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 nonfetid reddish-brownish mucus, 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 no fever (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 VD 5 and VD 3 (72.6%) cows resumed ovarian cyclicity. A smaller proportion of VD 5 (85.8%) cows received at least one 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. Milk production was 760 kg lower 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.

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).

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 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.

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.

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.

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.

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.

Effect of aluminized reflective hutch covers on calf health and performance.

The effect of polyethylene hutches wrapped in aluminized reflective covers (ARC) on health and performance of pre-weaned Holstein heifers during summer was evaluated. Ambient and hutch temperature and temperature-humidity index (THI) were also assessed. The study was conducted from June to October 2016 where temperature, humidity, and THI ranged from -1.3 to 38.4°C, 9.8 to 99.7%, and 33.6 to 81.1 units, respectively. Heifers enrolled at 1 d of life and housed in individual polyethylene hutches with (covered) or without (control) ARC were monitored until 60 d of life in 2 study groups (SG1, n = 94, monitored from June to September; and SG2, n = 101, monitored from August to October 2016). Calves were assessed twice per week for health status, behavior, rectal temperature, and respiratory rate. Similarly, hutch inner wall and sand bedding temperatures were determined for comparison between covered and control hutches housing calves. Four empty hutches per treatment group were continuously monitored for temperature and THI. The odds (95% confidence interval) of presenting diarrhea were 1.30 (1.01-1.60) times greater for calves housed in covered hutches than for those in the control group. Similarly, the odds of an abnormal ear score were 1.40 (1.03-2.00) times greater for calves in covered hutches compared with calves in control hutches. The odds of nasal discharge, eye discharge, and clinical dehydration did not differ between treatment groups. Rectal temperatures and respiratory rates were also similar in both groups. No differences were found in average daily gain between calves housed in covered and control hutches. Calf location determined at a fixed time during the day (starting at 1200 h) was associated with the use of ARC. The odds of remaining inside the hutch were 1.33 (1.03-1.70) times greater for calves housed in control compared with covered hutches. Overall, the average of hutch interior temperature was higher in covered compared with control hutches in both studies (SG1, 23.2 ± 0.06 vs. 22.8 ± 0.06°C; SG2, 17.1 ± 0.07 vs. 16.9 ± 0.07°C). Hutch THI was higher in covered empty hutches compared with control hutches in both study groups (SG1: 68.6 ± 0.06 vs. 67.6 ± 0.06°C; SG2: 60.2 ± 0.08 vs. 59.6 ± 0.08°C). Average temperature (measured at 1200 h) of the inner surface of the hutch wall was lower in covered compared with control hutches by 1°C (24.4 ± 0.13 vs. 25.4 ± 0.13°C), whereas the temperature of the sand bedding did not differ between groups. Our results suggest that the use of ARC did not generate a hutch microclimate that resulted in significant improvements on health and performance of pre-weaned dairy calves under these specific study settings.

Technical note: Evaluation of fine needle aspiration cytology for the diagnosis of fatty liver in dairy cattle.

Fatty liver is a common condition affecting dairy cattle during the periparturient period, characterized by a pathological accumulation of triglycerides (TG) in the hepatocytes. The objective of this study was to evaluate the diagnostic potential of fine needle aspiration cytology in fresh liver specimens using liver TG concentrations as a gold standard. Fifty-seven liver samples from Holstein cows were collected during processing at a slaughterhouse. Tissue and fine needle aspirate samples were obtained from the parietal upper portion of the caudate lobe. Two samples of liver tissue were collected with a 16 gauge × 15 cm biopsy needle for histological and TG concentration assessment. A third sample was collected for cytology using an 18 gauge × 5.08 cm needle. The contents of the needle were transferred to a glass slide, spread, and air-dried. Liver samples were assayed by colorimetry/fluorimetry to determine TG concentrations. Concentrations of TG <2% were considered normal. Histological and cytological evaluations were conducted by 2 different pathologists blind to the visual classification. Sensitivity (Se) and specificity (Sp) were calculated. Cytology had a Se and Sp of 73 and 85%, respectively. Histopathology had a Se and Sp of 45.9 and 100%, respectively. The likelihood of having higher scores for histopathology and cytology increased as a function of liver TG content (mg/g).

Season of conception is associated with future survival, fertility, and milk yield of Holstein cows.

Environmental influences during different stages of pregnancy can induce lifelong changes in the structure, physiology, and metabolism of the offspring. Our hypothesis was that season of conception (when the offspring was conceived), associated with heat stress conditions at conception and during the initial stages of embryonic development, affects the lifetime performance and survival of the female offspring after birth. The objective was to analyze the association between month of conception and subsequent survival, fertility, and milk yield in cows maintained on dairy farms in Florida, where the climate during the summer is hot and humid but winters are mild. Initial data consisted of 667,104 Dairy Herd Improvement lactation records from cows calving between 2000 and 2012 in 152 herds. Dates of conception were estimated as birth date minus 280 d. The magnitude of heat stress in each herd was quantified by comparing milk yield during summer and winter. Wood's lactation curves were fitted to adjust milk yields for effects of days in milk, and residuals were obtained for each calendar month. A sine function was fitted on the 12 residuals per farm. The difference between the highest and lowest points on the sine function was termed the seasonality index, a measure of the direct effect of heat stress on milk production. Herds were categorized in 3 levels of seasonality [low (seasonality index values less than the 25th percentile value; <2.84), medium (values within the interquartile range), and high (values greater than the 75th percentile value; >5.22)]. Cows were grouped by their month of conception: summer (July-September) and winter (December-February), and comparisons were performed by parity using logistic regression, ANOVA, and survival analysis. Two models were developed. Model A included the complete population of cows (n = 337,529 lactation records) conceived in winter or summer. Model B included cows (n = 228,257 lactation records) that had parent-average genetic information available to be able to correct for farmer's use of lower genetic merit of sires in summer. Other variables included in the models were month and year of calving, age at first calving, and herd. Models were run per parity group (1, 2, and ≥3). In both models, age at first calving was lower for cows conceived during winter versus summer. The odds (95% confidence interval) of survival to a second calving for cows conceived in winter were 1.21 and 1.15 times the odds of survival for cows conceived in summer for models A and B, respectively. Numbers of days from calving to first breeding and from calving to conception were consistently smaller for winter versus summer months of conception across all parity categories. Milk yields (305 d and by 70 d in milk) were greater for cows conceived in winter versus summer. In conclusion, cows that were conceived in the winter had better subsequent survival and performance than cows that were conceived in the summer. There is evidence that season of conception may have lifelong consequences for the offspring.

Retrospective evaluation of milk production and culling risk following either surgical, toggle-pin suture or conservative treatment of left displaced abomasum in Chilean dairy cows.

AIMS To describe milk yield and culling risk in cows diagnosed with left displacement of abomasum (LDA) treated either conservatively, by right flank pyloric omentopexy, or rolling and toggling, compared with normal herdmates from four Chilean dairy herds. METHODS Historical records were obtained from four commercial dairy farms located in Central Chile for cows with a history of LDA between 2010 and 2012, and healthy herdmates. Cows with LDA were categorised into three groups: cows treated with right omentopexy (ST, n=58), cows treated by toggle suturing (TT, n=15) and cows treated conservatively (CT, n=56). Control cows (n=129) were selected from unaffected cows, matched by days in milk (DIM), parity and herd with affected cows. Groups were compared for risk of culling up to 300 DIM and for milk production up to 5 months of lactation using survival and Cox proportional hazard models and mixed models for repeated measures, respectively. RESULTS Compared with cows in the Control group, the risk of being culled up to 300 DIM was 9.1 (SE 0.62) times greater in ST cows, 10.4 (SE 0.68) times greater in TT cows, and 37.3 (SE 0.61) times greater in CT cows (p<0.01). In the first 5 months of lactation, compared with cows in the Control group, mean daily milk production was 23.3 (SE 1.5) kg less in ST cows, 15.3 (SE 1.6) kg less in TT cows, and 30.1 (SE 1.3) kg less in CT cows (p<0.001). CONCLUSIONS AND CLINICAL RELEVANCE Cows in four dairy herds in central Chile diagnosed and treated for LDA produced significantly less milk and had a higher risk of culling than healthy herdmates. Although cows treated surgically or with toggle suture never recovered to the extent of healthy cows, they produced more milk than cows treated conservatively. However, the retrospective nature of the data, the inclusion of only four herds and the non-random allocation to treatments means that these conclusions cannot be extrapolated to the overall dairy cattle population in Chile.

Peripartal rumination dynamics and health status in cows calving in hot and cool seasons.

Our objective was to evaluate the effect of season of calving, associated with variable levels of heat stress, on the dynamics of rumination during the prepartum period and early lactation of cows that were healthy or affected by peripartal health disorders. Three weeks before the estimated due date, 210 multiparous Holstein cows at the University of Florida Dairy Unit were affixed with a neck collar containing rumination loggers, providing rumination time (RT) in 2-h periods. One blood sample was collected in a subpopulation of cows (n=76) at 12 to 48h postcalving to assess metabolic status by determining serum calcium, nonesterified fatty acid, and ß-hydroxybutyrate concentrations. The occurrence of peripartal health disorders (dystocia, clinical ketosis, clinical hypocalcemia, metritis, and mastitis) was assessed by University of Florida veterinarians and trained farm personnel. We analyzed the dynamics of daily RT over ± 14d relative to parturition in cows that were healthy or affected by specific health disorders by season of calving [hot season, June to September (n=77); cool season, November to April (n=118)] using repeated measures analysis and comparison of least squares means at different time points relative to calving. Rumination was consistently reduced on the day of calving in both healthy and sick cows in both the hot and cool seasons. Only hot-season calvings had shorter average daily RT prepartum and postpartum in cows affected by severe negative energy balance and subclinical ketosis. Dystocia during the hot season was associated with shorter daily RT prepartum; for cool-season calvings, cows with dystocia had reduced RT postpartum. We also observed reduced RT in cows with ketosis prepartum and postpartum in both the hot and cool seasons. Daily RT was reduced postpartum in cows with hypocalcemia and mastitis that calved during the cool season, and it was shorter in cows with metritis in both the hot and cool seasons. Our results indicated that the effect of heat stress on changes in rumination patterns around calving for sick cows depends on the specific health disorder or metabolic condition.

Effect of housing type on health and performance of preweaned dairy calves during summer in Florida.

The objective was to evaluate the effect of housing type on health and performance of preweaned dairy calves. One hundred calves, Holsteins (HO) or Holstein-Jersey crosses (HJ), were randomly assigned to a Calf-Tel (L. T. Hampel Corp., Germantown, WI) polyethylene hutch exposed to direct sun light (n=25 for both HO and HJ) or a wire hutch with a plywood shade located under trees (n=24 and 26 for HO and HJ, respectively). Calf rectal temperature, respiratory rate, health scores, and weaning weight were compared by housing type. Rectal temperatures were higher in calves in Calf-Tel hutches compared with calves in wire hutches at 1500 h, at 40.1±0.28 and 39.1±0.22°C, respectively. Similarly, respiratory rates were higher in calves in Calf-Tel hutches compared with calves in wire hutches at 1500 h, at 90±15 and 65±10 breaths per minute, respectively. Frequencies of calves presenting abnormal ear scores did not differ between calves in Calf-Tel or wire hutches. Abnormal eye scores were less likely to occur for calves in the wire hutches. Frequencies of calves with signs of respiratory disease, such as nasal discharge and coughing, were higher in the wire hutches compared with the Calf-Tel hutches. No differences were noted in the incidence and time of first diarrhea event in calves between hutch types. Calves in wire hutches had 5.47 greater odds of receiving veterinary treatment compared with calves in the Calf-Tel hutches. Weaning weights were not different for calves in Calf-Tel or in wire hutches. The lower frequency of calves with abnormal health scores and receiving veterinary treatment in Calf-Tel hutches and the lack of difference in weight gain suggests that this housing system adapted well to the specific environmental conditions of this study.

Effect of intrauterine infusion of an organic-certified product on uterine health, survival, and fertility of dairy cows with toxic puerperal metritis.

The objective was to evaluate the effect of intrauterine infusion of an organic certified product (Optimum UterFlush, Van Beek Natural Science, Orange City, IA) on uterine health, survival, and fertility of cows affected with toxic puerperal metritis (TPM) in an organic dairy farm. Cows with TPM were defined as having an abnormally enlarged uterus and a fetid watery red-brown vaginal discharge, associated with systemic illness and fever (rectal temperature ≥39.5°C), within 12 d postpartum. Cows diagnosed with TPM (n=220; study d 0) were blocked by parity (1 and ≥2) and randomly assigned into 1 of 2 intrauterine treatments applied every other day for a total of 3 applications: (1) control (CON)=200mL of povidone iodine diluted in 2L of distilled water (n=113); (2) Optimum UterFlush (UF)=3.75mL diluted in 117mL of distilled water (n=107). All enrolled cows received hypertonic saline solution (500mL 7.2% i.v.), dextrose (500mL 50% i.v.), and oral aspirin (5 boluses/d). Outcome variables for treatment efficacy included fever and presence of fetid vaginal discharge at study d 6 and 14, survival at study d 6 and 14 and at 30 and 100 DIM, and reproductive performance. Control variables were parity, BCS at enrollment, calving season, and milk yield. Occurrence of fever at d 6 and 14 was not different between the 2 treatment groups. Presence of fetid vaginal discharge at d 6 and 14 was lower in cows treated with UF compared with cows in the CON group (10.7 vs. 28% and 1.1 vs. 9.1%). The odds (95% confidence interval) for survival and remaining in the farm at study d 6 and 14 and at 30 and 100 DIM for cows in the UF treatment were 4.67 (1.38-15.8), 2.77 (1.25-6.10), 3.13 (1.22-8.02), and 2.82 (1.38-5.71) times the odds of cows in the CON group, respectively. The odds of AI until 150 DIM and the interval from calving to first AI were not different between the 2 treatments. However, pregnancy was affected by treatment; the odds (95% confidence interval) of pregnancy at the first AI, 150 DIM, and 300 DIM for cows treated with UF were 2.15 (1.05-4.40), 1.81 (1.04-3.15), and 1.92 (1.09-3.38) times the odds of cows in the CON group, respectively. Days to pregnancy were similar in both treatment groups, but the number of artificial inseminations per pregnancy were different (2.69 vs. 2.02 for cows in the CON and UF treatments). Results indicate that cows with TPM administered intrauterine infusion of Optimum UterFlush had higher odds of recovering and improved reproductive performance compared with cows treated with povidone iodine.