J. Dubuc

Risk factors for postpartum uterine diseases in dairy cows.

The objective of this observational study was to investigate the risk factors for metritis, purulent vaginal discharge, and cytological endometritis. The hypothesis was that purulent vaginal discharge and cytological endometritis would have different risk factors because they represent distinct manifestations of uterine disease. Data generated from 1,363 Holstein cows (3 herds) enrolled in a randomized clinical trial were used. Calving history, periparturient disease incidence, and body condition score at calving and at 63 d in milk (DIM) were recorded. Serum nonesterified fatty acid concentration was measured once during the week before expected calving. Serum nonesterified fatty acid, β-hydroxybutyric acid, and haptoglobin (Hapto) concentrations were measured at 4 ± 3, 11 ± 3, and 18 ± 3 DIM. Serum progesterone concentration was measured at 21 ± 3, 35 ± 3, 49 ± 3, and 63 ± 3 DIM. Metritis was diagnosed by farm managers within the first 20 DIM using a standardized definition. Cows were examined at 35 ± 3 DIM by a veterinarian for purulent vaginal discharge (mucopurulent or worse vaginal discharge; Metricheck device) and cytological endometritis (≥ 6% polymorphonuclear cells on endometrial cytology; cytobrush device). Statistical analyses were performed using multivariable logistic regression models for each disease, accounting for the random effect of herd. Risk factors for metritis included increased nonesterified fatty acid prepartum (≥ 0.6 mmol/L), dystocia, retained placenta, and increased Hapto in the first week postpartum (≥ 0.8 g/L). Risk factors for purulent vaginal discharge included twinning, dystocia, metritis, and increased Hapto (≥ 0.8 g/L) in the first week postpartum. Risk factors for cytological endometritis included low body condition score at parturition (≤ 2.75), hyperketonemia (≥ 1,100 μmol/L), and increased Hapto (≥ 0.8 g/L) in the first week postpartum. These results support the hypothesis that some of the risk factors for purulent vaginal discharge and cytological endometritis are different, which supports that they are distinct manifestations of uterine disease.

Definitions and diagnosis of postpartum endometritis in dairy cows

The objectives of this observational study were to determine and compare diagnostic criteria for postpartum endometritis in dairy cows. Data generated from 1,044 Holstein cows (6 herds) enrolled in a randomized clinical trial were used. Cows were examined for endometritis at 35±3 d (exam 1) and 56±3 d (exam 2) after parturition, using endometrial cytology (cytobrush technique), vaginal discharge scoring (Metricheck device; Simcrotech, Hamilton, New Zealand), and cervical diameter measurement (transrectal palpation). Reproductive data were recorded until 200 d after parturition. Diagnostic criteria for cytological and clinical endometritis were determined based on detrimental effect on subsequent reproductive performance, using logistic regression and Cox proportional hazard models accounting for the effect of herd clustering. Comparison of diagnostic criteria was performed using endometrial cytology as reference test or by quantifying the agreement between diagnostic approaches. At exam 1, diagnostic criteria were ≥6% polymorphonuclear cells and mucopurulent or worse (purulent or foul) vaginal discharge for cytological and clinical endometritis, respectively. At exam 2, diagnostic criteria were ≥4% polymorphonuclear cells and mucopurulent or worse vaginal discharge for cytological and clinical endometritis, respectively. Cows were classified as having cytological endometritis only, clinical endometritis only, or both cytological and clinical endometritis. Prevalence at exam 1 was 13.5, 9.4, and 5.8% for cytological endometritis only, clinical endometritis only, and both cytological and clinical endometritis, respectively. The detrimental effects of cytological and clinical endometritis on reproductive performance were additive. Among cows with clinical endometritis, only 38 and 36% had cytological endometritis at exam 1 and exam 2, respectively. Combination of diagnostic criteria improved neither the accuracy for predicting cytological endometritis nor the agreement between cytological and clinical endometritis. Overall, these results suggested that cytological and clinical endometritis may represent different manifestations of reproductive tract disease. They also suggested that use of the terminology clinical endometritis may not be accurate and that purulent vaginal discharge may be more descriptive.

Randomized clinical trial of antibiotic and prostaglandin treatments for uterine health and reproductive performance in dairy cows

The objectives of this study were to assess the efficacy of antibiotic treatment for preventing postpartum uterine disease among cows at high risk of uterine disease, and to assess the efficacy of PGF(2α) for treating cytological endometritis (CYTO) and purulent vaginal discharge (PVD). A total of 2,178 Holstein cows in 6 herds were enrolled in a randomized clinical trial. Within 24h after parturition, cows were classified at being at high risk of uterine disease (HRUD; n=1,017) if they had twins, dystocia, or retained placenta. All remaining cows were classified as being at low risk of uterine disease (LRUD; n=1,161). Cows in the HRUD group were randomly allocated in a factorial design to receive ceftiofur crystalline free acid (CCFA) at 24h after parturition or be untreated, and to receive dinoprost (PGF(2α)) at 35 and 49 (±3) days in milk (DIM) or to be untreated. Cows in LRUD were randomly allocated to receive PGF(2α) at 35 and 49 (±3) DIM or to be untreated. Serum progesterone was measured at 21, 35, 49, and 63 (±3) DIM. Cows were examined at 35 (±3; exam 1) and 56 (±3; exam 2) DIM for CYTO (by cytobrush device; ≥6% polymorphonuclear cells in endometrial cytology) and for PVD (by Metricheck device; mucopurulent or purulent vaginal discharge). Statistical analyses were performed using multivariable logistic regression models accounting for herd clustering. Treatment with CCFA in HRUD cows was not associated with the probability of metritis overall, but interactions occurred such that CCFA decreased the incidence of metritis among HRUD cows that did not have retained placenta and among cows of parity ≥2. Treatment with CCFA in HRUD cows decreased the probability of PVD at exam 1. Treatment with PGF(2α) did not affect the probability of cure of CYTO or PVD irrespective of progesterone concentration at the time of treatment. Among cows affected by CYTO or PVD at exam 1, 66 and 63%, respectively, had spontaneously cured at exam 2. Cows persistently affected at exam 2 had an increased time to pregnancy and were more likely to have both CYTO and PVD at exam 1. Administration of PGF(2α) at both 5 and 7 wk postpartum did not mitigate the effects of CYTO or PVD on reproductive performance. Clinical approaches to treatment of chronic postpartum reproductive tract infection and inflammation should be reassessed.

Effects of postpartum uterine diseases on milk production and culling in dairy cows

The objective was to quantify the effect of postpartum uterine diseases on milk production and culling. Data from 2,178 Holstein cows in 6 herds enrolled in a randomized clinical trial were used. Milk production data from the first 4 Dairy Herd Improvement Association (DHIA) test-days and culling data from farm records were collected. Retained placenta (RP; ≥24 h after parturition) and metritis [≤20 d in milk (DIM)] were diagnosed by farm managers using standardized definitions. Farms were visited weekly and cows were examined at 35 and 56 (±3) DIM using endometrial cytology (cytobrush device), vaginal discharge scoring (Metricheck device), and measurement of cervical diameter by transrectal palpation. Diagnostic criteria for cytological endometritis (CYTO) and purulent vaginal discharge (PVD) were established based on a detrimental effect on subsequent reproduction. Statistical analyses were performed using linear mixed models, logistic regression models, and Cox proportional hazard models, accounting for the effects of experimental treatments and herd clustering. Milk production and culling were the outcomes. Primiparous and multiparous cows were modeled separately for milk production. Milk production of primiparous cows was unaffected by uterine diseases. The effect of metritis on milk production was variable over time in multiparous cows: it decreased production per cow by 3.7 kg at the first DHIA test, but was not different at later tests. Retained placenta decreased milk production by 2.6 kg/d in multiparous cows through the first 4 DHIA tests. The projected effects of metritis and RP in multiparous cows were reductions of 259 kg and 753 kg over 305 DIM, respectively; these effects were additive. Neither CYTO nor PVD affected milk production. Culling risks at 30 and 63 DIM were unaffected by RP and metritis. Culling hazard up to 300 DIM was unaffected by RP, metritis, CYTO, or PVD, whether or not pregnancy status, milk production, and displaced abomasum were accounted for. Uterine disease decreased pregnancy rate, which was a substantial risk factor for culling; however, if affected cows became pregnant they were not at greater risk of culling.

Metabolic parameters in transition cows as indicators for early-lactation culling risk

A retrospective cohort study was performed with the objective of determining whether the serum concentrations of nonesterified fatty acids (NEFA), β-hydroxybutyric acid (BHBA), or calcium were associated with the risk of culling within 60 d in milk (DIM) in Holstein cows, and to establish thresholds for each metabolite that were predictive of increased culling risk. Data from 5,979 cows in Ontario (Canada) and several US states were obtained from 4 previously reported studies. For each metabolite and each of 3 sampling weeks (-1, +1, and +2 relative to calving), an optimal threshold was calculated based on having the maximum combined sensitivity (Se) and specificity (Sp) and used to categorize the serum concentrations into high and low risk groups. Logistic regression models were built for each metabolite and each week of sampling, as well as considering together all metabolites in wk -1 and wk +1 relative to calving. Cow was considered the experimental unit and herd as a random effect. Considered separately, precalving NEFA ≥ 0.4 mmol/L [odds ratio (OR)=1.8; 95% confidence interval (CI)=1.4 to 2.2], NEFA ≥ 0.8 mmol/L in wk +1 relative to calving (OR=2.0; 95% CI=1.5 to 2.6) and NEFA ≥ 0.8 mmol/L in wk +2 (OR=4.2; 95% CI=1.9 to 9.4 for cows in lactation 2; OR=2.1; 95% CI=1.4 to 3.3 for cows in lactation ≥ 3) were each associated with an increased risk of culling within the first 60 DIM. Similarly, BHBA ≥ 0.7 mmol/L in wk -1 (OR=1.8; 95% CI=1.3 to 2.5), BHBA ≥ 1.2 mmol/L in wk +1 (OR=1.8; 95% CI=1.4 to 2.2), and BHBA ≥ 1.6 mmol/L in wk +2 (OR=3.2; 95% CI=1.6 to 6.4 for cows in lactation 2; OR=2.3; 95% CI=1.6 to 3.3 for cows in lactation ≥ 3) were each associated with an increased risk of culling within the first 60 DIM. Likewise, calcium ≤ 2.3 mmol/L in wk -1 (OR=1.6; 95% CI=1.2 to 2.2), calcium ≤ 2.2 mmol/L in wk +1 (OR=1.5; 95% CI=1.2 to 1.9), and calcium ≤ 2.3 mmol/L in wk +2 (OR=2.3; 95% CI=1.1 to 3.1) were each associated with an increased risk of culling within the first 60 DIM. When all metabolites were analyzed together, serum NEFA and calcium concentrations in wk -1 and serum NEFA concentration in wk +1 remained in the models. In conclusion, elevated serum NEFA and BHBA concentrations and lower serum calcium concentrations within 1 wk before calving through 2 wk after calving were associated with an increased risk of culling in early lactation. Measuring the concentration of selected metabolites around parturition may help to develop monitoring and intervention strategies to prevent early culling in transition dairy cows.

Reproductive tract defense and disease in postpartum dairy cows

This paper briefly reviews recent data and concepts on the development and mitigation of infection and inflammation in the reproductive tract of dairy cows during the first 2 mo after calving. The incidence of metritis is typically between 10 and 20%, of clinical endometritis or purulent vaginal discharge (PVD) approximately 15%, and of subclinical or cytological endometritis a further 15%. Worse postpartum negative energy balance is associated with more severe or prolonged uterine inflammation. Changes in feed intake, expression of genes for pro-inflammatory cytokines, notably interleukin (IL) 1, IL6 and IL8, circulating concentrations of beta-hydroxybutyrate (BHBA) or nonesterified fatty acids (NEFA), and innate immune function precede both metritis and endometritis by several weeks. Infections with Escherichia coli and Arcanobacterium pyogenes are associated with both metritis and PVD. There are new data to suggest that specific virulence factors in E. coli associated with adherence may be important in metritis and PVD. Cytological endometritis and PVD are overlapping but largely distinct conditions, and there are emerging data that cervicitis exists both concurrent with and separate from endometritis. Much remains to be learned about what initiates and sustains harmful inflammation of the reproductive tract. Such information is necessary to develop effective treatments for the various forms of disease and, more importantly, to develop means to prevent endometritis and cervicitis. In particular, vaccination against specific uterine pathogens and interventions to modulate innate immune response appear to be important avenues for investigation. Presently, commonly recommended best management practices for cows in the transition period are likely to be helpful to mitigate the risk of reproductive disease.

Risk factors and effects of postpartum anovulation in dairy cows

The objectives were to identify risk factors for and to quantify the effect of postpartum anovulation on reproductive performance in dairy cows. Data from 2,178 Holstein cows in 6 commercial herds enrolled in a randomized clinical trial were used. Data on periparturient disease incidence, calving history, and body condition score were collected. Cows were examined at wk 5 postpartum for reproductive tract disease; cytological endometritis was defined as ≥6% polymorphonuclear cells in endometrial cytology, and purulent vaginal discharge was defined as the presence of mucopurulent or purulent vaginal discharge. Cows were followed until 300 d in milk (DIM) for reproductive performance. Serum nonesterified fatty acids (NEFA) concentration was measured once during the week before expected calving. Serum β-hydroxybutyrate, NEFA, and haptoglobin were measured at wk 1, 2, and 3 postpartum. Serum progesterone (P4) was measured at wk 3, 5, 7, and 9 postpartum. The end of the postpartum anovulation period was defined as the first sampling time at which P4 was >1 ng/mL. Statistical analyses were performed using logistic regression models and Cox proportional hazard models. The prevalence of anovulation was 72, 44, 26, and 17% at wk 3, 5, 7, and 9, respectively. Cows were classified according to their ovulatory status as having luteal function at 21 DIM (Cyc21), as having low P4 at 21 DIM but having luteal function at least once at 35 or 49 or 63 DIM (Cyc63), or being anovulatory at 63 DIM (Anov63; no samples with P4 >1 ng/mL). Factors associated with early ovulation (Cyc21) included season, parity, decreased haptoglobinemia, and decreased serum NEFA concentration before and after parturition. Risk factors for prolonged anovulation (Anov63) included cytological endometritis, increased haptoglobinemia, and greater serum NEFA concentrations before and after parturition. Cows classified as Anov63 had an increased median time to first breeding compared with Cyc63 (74.1 vs. 73.2 d). The effect of prolonged postpartum anovulation on median time to pregnancy was conditional on parity group; a detrimental effect was present in cows of parity ≥3 (129 d for Cyc21, 151 d for Cyc63, and 180 d for Anov63), but no effect was observed in cows of parity ≤2. Overall, these findings suggest that postpartum anovulation was associated with indicators of energy balance and uterine inflammation, and with detrimental effects on reproductive performance.

Accuracy of milk ketone bodies from flow-injection analysis for the diagnosis of hyperketonemia in dairy cows.

The objectives of this study were (1) to determine the correlations between blood β-hydroxybutyrate (BHBA) and milk components [BHBA, acetone, fat, protein, and fat:protein (F:P) ratio], and (2) to establish optimal thresholds for milk components to predict hyperketonemia in dairy cows. Data on 163 cows from 37 herds were used in this cross-sectional study. Herds were visited once during the study period, and cows between 2 and 90 d in milk were blood sampled within 4h of milk sampling for the Dairy Herd Improvement test. Blood BHBA concentrations were measured using a cow-side electronic meter, Precision Xtra, which was considered the gold standard test in this study. Milk BHBA and acetone concentrations were measured in Dairy Herd Improvement milk samples by flow-injection analysis; whereas, milk fat and protein were tested using Fourier transform infrared spectroscopy. Hyperketonemia was defined by a blood BHBA concentration ≥ 1.4 mmol/L. The prevalence of hyperketonemia (based on blood BHBA values) in this study population was 21.0%. Pearson correlation coefficients between blood BHBA and milk BHBA, acetone, fat, protein, and F:P ratio were 0.89, 0.73, 0.21, 0.04, and 0.17, respectively. Receiver operating characteristic curves were generated and thresholds for each individual milk component were determined based on the maximal sum of sensitivity and specificity. Optimal threshold values for hyperketonemia were milk BHBA ≥ 0.20 mmol/L, acetone ≥ 0.08 mmol/L, fat ≥ 4.2%, and F:P ratio ≥ 1.3. Based on these thresholds, milk BHBA and acetone had greater sensitivity (84 and 87%, respectively) and greater specificity (96 and 95%, respectively) than the other milk components (fat, protein, and F:P). Series and parallel testing slightly improved the accuracy of milk BHBA and acetone values to predict hyperketonemia. A multivariable model that accounted for milk BHBA and milk acetone values simultaneously had the highest accuracy of all tested models for predicting hyperketonemia. These results support that milk BHBA and milk acetone values from flow-injection analysis are accurate diagnostic tools for hyperketonemia in dairy cows and could potentially be used for herd-level hyperketonemia surveillance programs.

Determination of optimal diagnostic criteria for purulent vaginal discharge and cytological endometritis in dairy cows.

The objectives of this observational study were to identify the optimal diagnostic criteria for purulent vaginal discharge (PVD) and cytological endometritis (ENDO) using vaginal discharge, endometrial cytology, and leukocyte esterase (LE) tests, and to quantify their effect on subsequent reproductive performance. Data generated from 1,099 untreated Holstein cows (28 herds) enrolled in a randomized clinical trial were used in this study. Cows were examined at 35 (± 7) d in milk for PVD using vaginal discharge scoring and for ENDO using endometrial cytology and LE testing. Optimal combinations of diagnostic criteria were determined based on the lowest Akaike information criterion (AIC) to predict pregnancy status at first service. Once identified, these criteria were used to quantify the effect of PVD and ENDO on pregnancy risk at first service and on pregnancy hazard until 200 d in milk (survival analysis). Predicting ability of these diagnostic criteria was determined using area under the curve (AUC) values. The prevalence of PVD and ENDO was calculated as well as the agreement between endometrial cytology and LE. The optimal diagnostic criteria (lowest AIC) identified in this study were purulent vaginal discharge or worse (≥ 4), ≥ 6% polymorphonuclear leukocytes (PMNL) by endometrial cytology, and small amounts of leukocytes or worse (≥ 1) by LE testing. When using the combination of vaginal discharge and PMNL percentage as diagnostic tools (n = 1,099), the prevalences of PVD and ENDO were 17.1 and 36.2%, respectively. When using the combination of vaginal discharge and LE (n = 915), the prevalences of PVD and ENDO were 17.1 and 48.4%. The optimal strategies for predicting pregnancy status at first service were the use of LE only (AUC = 0.578) and PMNL percentage only (AUC = 0.575). Cows affected by PVD and ENDO had 0.36 and 0.32 times the odds, respectively, of being pregnant at first service when using PMNL percentage compared with that of unaffected cows; odds ratios were 0.33 and 0.69 for PVD and ENDO, respectively, when LE was used. Kappa value (agreement) of the pairwise comparison for the PMNL percentage and LE was 0.43. Vaginal discharge, endometrial cytology, and LE can be used to diagnose PVD and ENDO, and to predict pregnancy status at first service. The use of LE could be a good alternative to endometrial cytology for on-farm testing.

A randomized herd-level field study of dietary interactions with monensin on milk fat percentage in dairy cows.

This field trial evaluated the effects of dietary supplementation with 16 mg/kg (based on total dry matter intake) of monensin sodium on bulk tank milk fat percentage (MFP) of commercial dairy herds. Interactions between monensin and nutritional factors on MFP were studied. The trial was conducted in 47 Holstein dairy herds in Québec, Canada, between November 2005 and May 2006. The herd was the unit of interest. Enrolled herds were followed for a 7-mo period. Monensin treatment was randomly allocated in a crossover design where monensin was supplemented to the lactating dairy cow diet for a consecutive 12-wk period. Twenty-four herds were allocated to monensin treatment for the first period of trial, and 23 herds were allocated for the second period. Diet composition and ration physically effective particle level were collected every 8 wk. Milk fat percentage data were retrieved from weekly bulk tank measures. Data were analyzed in linear mixed models using repeated measures within herd where MFP was considered the outcome variable. In addition to the main effect of monensin treatment, the following covariates were forced a priori into all statistical models: treatment period, weekly herd mean parity, and weekly herd mean days in milk. The majority of herds were fed a total mixed ration (n = 29) and were housed in tie-stalls (n = 42). Monensin significantly decreased bulk tank MFP by 0.12 percentage points. The reduction of MFP associated with monensin was larger for herds having a diet high (>39.7%) in nonfiber carbohydrates, having a low level of physically effective particles in ration (>45.0%; >or=8 mm), and not feeding dry hay as first meal in the morning. Significant interactions between monensin and nutritional factors on bulk tank MFP were related to nonfiber carbohydrate and fiber concentrations in the diet.