K.E. Leslie

Impact of hyperketonemia in early lactation dairy cows on health and production

Data from 1,010 lactating lactating, predominately component-fed Holstein cattle from 25 predominately tie-stall dairy farms in southwest Ontario were used to identify objective thresholds for defining hyperketonemia in lactating dairy cattle based on negative impacts on cow health, milk production, or both. Serum samples obtained during wk 1 and 2 postpartum and analyzed for beta-hydroxybutyrate (BHBA) concentrations that were used in analysis. Data were time-ordered so that the serum samples were obtained at least 1 d before the disease or milk recording events. Serum BHBA cutpoints were constructed at 200 micromol/L intervals between 600 and 2,000 micromol/L. Critical cutpoints for the health analysis were determined based on the threshold having the greatest sum of sensitivity and specificity for predicting the disease occurrence. For the production outcomes, models for first test day milk yield, milk fat, and milk protein percentage were constructed including covariates of parity, precalving body condition score, season of calving, test day linear score, and the random effect of herd. Each cutpoint was tested in these models to determine the threshold with the greatest impact and least risk of a type 1 error. Serum BHBA concentrations at or above 1,200 micromol/L in the first week following calving were associated with increased risks of subsequent displaced abomasum [odds ratio (OR) = 2.60] and metritis (OR = 3.35), whereas the critical threshold of BHBA in wk 2 postpartum on the risk of abomasal displacement was >or=1,800 micromol/L (OR = 6.22). The best threshold for predicting subsequent risk of clinical ketosis from serum obtained during wk 1 and wk 2 postpartum was 1,400 micromol/L of BHBA (OR = 4.25 and 5.98, respectively). There was no association between clinical mastitis and elevated serum BHBA in wk 1 or 2 postpartum, and there was no association between wk 2 BHBA and risk of metritis. Greater serum BHBA measured during the first and second week postcalving were associated with less milk yield, greater milk fat percentage, and less milk protein percentage on the first Dairy Herd Improvement test day of lactation. Impacts on first Dairy Herd Improvement test milk yield began at BHBA >or=1,200 micromol/L for wk 1 samples and >or=1,400 micromol/L for wk 2 samples. The greatest impact on yield occurred at 1,400 micromol/L (-1.88 kg/d) and 2,000 micromol/L (-3.3 kg/d) for sera from the first and second week postcalving, respectively. Hyperketonemia can be defined at 1,400 micromol/L of BHBA and in the first 2 wk postpartum increases disease risk and results in substantial loss of milk yield in early lactation.

Evaluation of changes in somatic cell counts as indicators of new intramammary infections

Data from 1565 milk samples collected weekly from the University of Guelph mastitis research herd were used to evaluate somatic cell counts (SCC) as a screening test for prevalent intramammary infections. Changes in SCC were evaluated as a screening test for incident infections. The sensitivity and specificity of SCC for prevalent infections depended on whether cows with only minor pathogen infections were classified as infected or not. Including them in the infected cow category resulted in estimates of sensitivity and specificity of 0.726 and 0.855, respectively, at a threshold of 200 000 cells ml−1. Several methods of masuring the change in a cows cell count over a 28-day interval were evaluated. Observing a change from under to over a threshold of 200 000 cells ml−1 was optimal for predicting new infections. While the sensitivity of this procedure was low (0.388), the specificity was high (0.919) and relatively few cows (0.126) which were already infected at the start of the interval met the criterion. This last value was referred to as the TPNAR (test positive, not at risk) and was required, along with the sensitivity and specificity, to completely define the operating characteristics of the test and to predict the expected performance of the test. Changes in the apparent 28-day incidence risk in a herd would be expected to be much smaller than changes in the true risk. The correlation between the observed incidence risk in the study herd over a number of test periods ranged from 0.57 to 0.83.

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.

RELATIONSHIP BETWEEN MATERNAL PLASMA PROGESTERONE CONCENTRATION AND INTERFERON-TAU SYNTHESIS BY THE CONCEPTUS IN CATTLE

The objective of this study was to determine the relationship between maternal progesterone concentration and conceptus synthesis of interferon-tau as an index of conceptus viability at the time of maternal recognition of pregnancy. Heifers of mixed beef breeds were randomly assigned to receive 1 of 2 treatments: 1) intramuscular injection of 1500 IU hCG on Day 5 after artificial insemination (AI; n = 12) or 2) intramuscular injection of saline on Day 5 after AI (n = 17). Ovaries were scanned daily by transrectal real-time ultrasonography. Progesterone concentrations were determined from daily blood samples collected from the jugular vein. Heifers were slaughtered on Day 18 after AI and conceptus tissues were collected. These were incubated individually at 37 degrees C in RPMI medium, and supernatant collected after 24 h. Conceptus secretory products in the supernatant were analyzed for interferon concentration by antiviral assay using vesicular stomatitis virus. Transrectal ultrasonography showed all heifers that received hCG had at least 1 extra corpus luteum (CL) in addition to the spontaneous CL formed from the previous ovulation (10 with 2 CL, 2 with 3 CL). A significant increase in plasma progesterone concentration was detected in pregnant heifers treated with hCG (n = 9) vs pregnant control heifers (n = 11; P < 0.001). There was a tendency for an increase (P = 0.059) in synthesis of interferon-tau by conceptuses from hCG-treated heifers compared to control heifers. Maternal plasma progesterone concentrations were correlated with interferon-tau production by the conceptuses (r = 0.593, P < 0.006), suggesting that higher maternal progesterone may provide a more suitable environment for the developing conceptus.

Factors associated with morbidity, mortality, and growth of dairy heifer calves up to 3 months of age

Calfhood disease is an important problem on many dairy operations that can have substantial effects on heifer survival and productivity, and has economic and welfare impacts. Neonatal calf diarrhea (NCD) and bovine respiratory disease (BRD) are the most common causes of morbidity and mortality in young dairy cattle. The objective of this observational study was to investigate factors associated with the risks of morbidity and mortality, and with growth, in commercial dairy heifers calves. A total of 2874 heifer calves from 19 commercial dairy farms in Minnesota and Ontario were enrolled at 1-7 days of age and followed for approximately 3 months. Using cut-points of serum total protein of 5.2 and 5.7 g/dl, the incidences of failure of transfer of passive immunity (FTPI) were 11 and 32%, respectively. A cut-point of 5.7 g/dl was the most predictive of BRD before 5 weeks of age (sensitivity=40%, specificity=69%). The positive predictive value was poor (PPV=18%), but the negative predictive value was good (NPV=87%). A cut-point of 5.2g/dl was most predictive of death before 5 weeks of age (sensitivity=27%, specificity=89%, PPV=5%, NPV=98%). Serum total protein during the first week of life was a poor predictor of NCD. Over 23% of calves were treated for diarrhea. Risk factors were weight at enrollment, other diseases before 2 weeks of age, and an interaction between season of birth and herd-level incidence of NCD. Almost 22% of calves were treated at least once for BRD. Factors associated with an increased risk of BRD included herd-level incidence of BRD, season of birth, navel dipping, other diseases before 2 weeks of age, failure of transfer of passive immunity, and manual control of temperature in pre-weaning housing. Administration of supplemental antibody products at birth was associated with a reduced incidence of BRD. Overall mortality was 3.5%. Risk of mortality was increased by treatment for BRD and other diseases. The mean average weight gain was 0.95 kg/day (range: 0.11-1.62 kg/day; SD=0.2). Twinning status, FTP, treatment for NCD or other diseases, and month of birth influenced body weight. This study illustrated relationships among various diseases, mortality, and growth. Furthermore, it demonstrated the importance of colostrum for protection against BRD and improved growth performance, while bringing into question the optimal method of determining failure of transfer of passive immunity.

The selective treatment of clinical mastitis based on on-farm culture results: I. Effects on antibiotic use, milk withholding time, and short-term clinical and bacteriological outcomes.

The objective of this multi-state, multi-herd clinical trial was to evaluate the efficacy of using an on-farm culture system to guide strategic treatment decisions in cows with clinical mastitis. The study was conducted in 8 commercial dairy farms ranging in size from 144 to 1,795 cows from Minnesota, Wisconsin, and Ontario, Canada. A total of 422 cows affected with mild or moderate clinical mastitis in 449 quarters were randomly assigned to either (1) a positive-control treatment program or (2) an on-farm, culture-based treatment program. Quarter cases assigned to the positive-control group received immediate on-label intramammary treatment with cephapirin sodium. Quarters assigned to the culture-based treatment program were cultured on-farm and treated with cephapirin sodium after 18 to 24h of incubation if they had gram-positive growth or a mixed infection. Quarters with gram-negative or no growth did not receive intramammary therapy. The proportion of quarter cases assigned to positive-control and culture-based treatments that received intramammary antibiotic therapy because of study assignment was 100 and 44%, respectively; the proportion of cases that received secondary antibiotic therapy was 36 and 19%, respectively; and the proportion of cases that received intramammary antibiotic therapy because of study assignment or secondary therapy was 100 and 51%, respectively. A tendency existed for a decrease in the number of days in which milk was discarded from cows assigned to the culture-based treatment program versus cows assigned to the positive-control group (5.9 vs. 5.2 d). No statistically significant differences existed between cases assigned to the positive-control and cases assigned to the culture-based treatment program in days to clinical cure (2.7 vs. 3.2 d), bacteriological cure risk within 21 d of enrollment (71 vs. 60%), new intramammary infection risk within 21 d of enrollment (50 vs. 50%), and treatment failure risk (presence of infection, secondary treatment, clinical mastitis recurrence, or removal from herd within 21 d after enrollment; 81 vs. 78%). In summary, the use of an on-farm culture system to guide the strategic treatment of clinical mastitis reduced intramammary antibiotic use by half and tended to decrease milk withholding time by 1 d, without significant differences in days to clinical cure, bacteriological cure risk, new intramammary infection risk, and treatment failure risk within 21 d after the clinical mastitis event.

Efficacy and Pharmacokinetics of Bacteriophage Therapy in Treatment of Subclinical Staphylococcus aureus Mastitis in Lactating Dairy Cattle

ABSTRACT Bovine mastitis is an inflammation of the udder caused by microbial infection. Mastitis caused by Staphylococcus aureus is a major concern to the dairy industry due to its resistance to antibiotic treatment and its propensity to recur chronically. Growing concerns surrounding antibiotic resistance have spurred research into alternative treatment methods. The ability of lytic S. aureus bacteriophage K to eliminate bovine S. aureus intramammary infection during lactation was evaluated in a placebo-controlled, multisite trial. Twenty-four lactating Holstein cows with preexisting subclinical S. aureus mastitis were treated. Treatment consisted of 10-ml intramammary infusions of either 1.25 × 1011 PFU of phage K or saline, administered once per day for 5 days. The cure rate was established by the assessment of four serial samples collected following treatment. The cure rate was 3 of 18 quarters (16.7%) in the phage-treated group, while none of the 20 saline-treated quarters were cured. This difference was not statistically significant. The effects of phage intramammary infusion on the bovine mammary gland were also studied. In healthy lactating cows, a single infusion of either filter-sterilized broth lysate or a CsCl gradient-purified phage preparation elicited a large increase in the milk somatic cell count. This response was not observed when phage was infused into quarters which were already infected with S. aureus. Phage-infused healthy quarters continued to shed viable bacteriophage into the milk for up to 36 h postinfusion. The phage concentration in the milk suggested that there was significant degradation or inactivation of the infused phage within the gland.

The association of serum metabolites in the transition period with milk production and early-lactation reproductive performance

The objective was to examine the associations of peripartum concentrations of nonesterified fatty acids (NEFA), β-hydroxybutyrate (BHBA), and calcium with milk production in early lactation and pregnancy at the first artificial insemination (AI) across different management systems. Fifty-five Holstein freestall dairy herds located across the United States and Canada were visited weekly for blood sample collection from 2,365 cows. For each week of sampling (from wk -1 through wk 3 relative to calving) and for each metabolite, serum concentrations were dichotomized at various thresholds to identify the thresholds with the best negative associations with milk production and pregnancy at first AI. These thresholds were used to categorize the serum concentrations into higher and lower risk categories. Repeated-measures ANOVA and multivariable logistic regression were conducted for milk production and pregnancy at the first AI data, respectively, considering cow as the experimental unit and herd as a random effect. In the week before calving, serum NEFA ≥ 0.5 mEq/L, BHBA ≥ 600 μmol/L, and calcium ≤ 2.1 mmol/L were associated with 1.6 to 3.2 kg/d milk loss across the first 4 Dairy Herd Improvement Association (DHIA) milk tests. High levels of NEFA and BHBA in wk 1 and 2 after calving (≥ 0.7 and ≥ 1.0 mEq/L for NEFA, and ≥ 1,400 and ≥ 1,200 μmol/L for BHBA), and low levels of calcium (≤ 2.1 mmol/L) in wk 1, 2 and 3 after calving were associated with milk loss at the first DHIA milk test. Serum concentrations of NEFA and BHBA were not associated with pregnancy at first AI in any sampling week, whereas calcium <2.2 to 2.4 mmol/L from wk 1 through wk 3 postpartum were associated with reduced pregnancy at first AI. In conclusion, high serum concentrations of NEFA, BHBA, and low concentrations of calcium around parturition were associated with early lactation milk loss, and low calcium concentration around parturition was associated with impaired early lactation reproduction.

An evaluation of Brix refractometry instruments for measurement of colostrum quality in dairy cattle.

Acquisition of high quality colostrum is an important factor influencing neonatal calf health. Many methods have been used to assess the Ig concentration of colostrum; however, improved, validated evaluation tools are needed. The aims of this study were to evaluate both optical and digital Brix refractometer instruments for the measurement of Ig concentration of colostrum as compared with the gold standard radial immunodiffusion assay laboratory assessment and to determine the correlation between Ig measurements taken from fresh and frozen colostrum samples for both Brix refractometer instruments. This research was completed using 288 colostrum samples from 3 different farms. It was concluded that the optical and digital Brix refractometers were highly correlated for both fresh and frozen samples (r=0.98 and r=0.97, respectively). Correlation between both refractometer instruments for fresh and frozen samples and the gold standard radial immunodiffusion assay were determined to be very similar, with a correlation coefficient between 0.71 and 0.74. Both instruments exhibited excellent test characteristics, indicating an appropriate cut-off point of 22% Brix score for the identification of good quality colostrum.