Economic comparison between ceftiofur-treated and nontreated dairy cows with metritis

The objective was to investigate the economic effect of treating dairy cows with metritis using ceftiofur-free acid or leaving them untreated at the time of diagnosis. Cows with a fetid, watery, red-brownish vaginal discharge were diagnosed with metritis (d 0). Data from 875 dairy cows (506 primiparous and 369 multiparous) from 1 herd in northern Florida that had been part of a larger study evaluating different treatments for metritis were used for the economic analysis. Holstein cows with metritis had been randomly assigned to: Ceftiofur (CEF, n = 239) = subcutaneous injection of 6.6 mg/kg of ceftiofur crystalline-free acid in the base of the ear at d 0 and d 3; Untreated (UNT, n = 233) = no treatment applied at metritis diagnosis. Both groups could receive escape therapy if condition worsened. A group of nonmetritic healthy cows (NMET; n = 403) from the same cohort was randomly selected for comparison. Continuous outcomes such as 300-d milk production (kg/cow), milk sales ($/cow), cow sales ($/cow), treatment cost by 60 days in milk ($/cow), reproduction cost ($/cow), replacement cost ($/cow), feeding cost ($/cow), and gross profit per cow ($/cow) were analyzed using the ANOVA (MIXED procedure of SAS version 9.4). Dichotomous outcomes such as pregnancy and culling by 300 d were analyzed using logistic regression (GLIMMIX procedure of SAS). Models included the fixed effects of treatment, parity, and the interaction between treatment and parity. A stochastic analysis was performed with 10,000 iterations using the observed results from each group. The CEF treatment resulted in greater treatment cost by 60 DIM than UNT ($112 vs. $37), but resulted in a greater proportion of pregnant cows (71 vs. 61%) and decreased culling by 300 DIM (29 vs. 39%) compared with UNT. Gross profit was lesser for UNT than NMET ($2,969 vs. $3,426), and CEF was intermediate ($3,219). The stochastic analysis showed that the mean difference in gross profit between UNT and NMET was ?$457; saleable milk (49%) and replacement cost (24%) accounted for most of the variation. The mean difference in gross profit between CEF and NMET group was ?$207; saleable milk (82%) and initial metritis treatment cost (9%) accounted for most of the variation. The mean difference in gross profit between the UNT and the CEF group was ?$250; replacement cost (41%) and cow sales (31%) accounted for most of the variation. In summary, metritis caused large economic losses when left untreated, and CEF reduced those losses by improving fertility, reducing culling and replacement cost, and reducing milk yield losses.     

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.     

Feeding behavior identifies dairy cows at risk for metritis

Dairy cows experience a high incidence of disease and metabolic disorders in the weeks immediately following calving, but early and accurate diagnosis remains a challenge. Cows suffering from metritis, one common disease following calving, exhibit reduced milk yield and reproductive performance. However, afflicted cows show few overt signs of illness and frequently go unnoticed in the absence of veterinary examination. To determine whether changes in feeding behavior could be used to identify animals at risk for metritis, attendance at the feed alley was monitored continuously for 26 Holstein cows during the transition period, beginning 2 wk before and ending 3 wk after calving. Every 3 ± 1 d, cows were examined for metritis based on rectal body temperature and condition of vaginal discharge. Over the 3 wk of observations after calving, 69% of cows showed some signs of metritis. These cows spent on average 22 min/d less time at the feed alley during the transition period than did non-metritic cows. For every 10-min decrease in average daily feeding time, cows were twice as likely to be diagnosed with metritis. A threshold of 75 min of average daily feeding time was 89% sensitive and 62% specific for detection of acute metritis. In conclusion, reduced time at the feeder can be used to identify dairy cows at risk for metritis. More research is required to determine how soon before calving at-risk cows can be identified and whether these behavioral differences can also be used in the early diagnosis of other diseases or metabolic disorders.     

Efficacy of ampicillin trihydrate or ceftiofur hydrochloride for treatment of metritis and subsequent fertility in dairy cows

Our objectives were to evaluate the efficacy of ampicillin trihydrate for the treatment of metritis in dairy cows compared with ceftiofur hydrochloride and the subsequent effects on pregnancy at first insemination (P/AI). Cows in the first 12 d in milk (DIM) with a uterine discharge score of 5 (watery, reddish or brownish discharge of foul smell) and rectal temperature <39.5°C were diagnosed with metritis based on the fetid discharge, and cows with metritis and rectal temperature ≥39.5°C were diagnosed as having puerperal metritis. Cows with metritis (n = 528) were blocked by parity and type of metritis as fetid discharge or puerperal metritis and, within each block, assigned randomly to receive 11 mg/kg of ampicillin (n = 259) or 2.2 mg/kg of ceftiofur (n = 269) once daily for 5 d. Day of diagnosis of metritis was considered study d 1. A cohort of 268 cows without metritis was selected randomly at 12 DIM. Rectal temperature was measured in cows with metritis on study d 1 to 7, and 12, and vaginal discharge was scored on study d 5, 7, and 12. Metritis cure was characterized by vaginal discharge score of <5 or by vaginal discharge score of <5 and no fever. At 32 ± 3 DIM, vaginal discharge was scored for diagnosis of purulent vaginal discharge. At 39 ± 3 DIM, endometrial cytology was performed. At 53 ± 3 and 67 ± 3 DIM, ovaries were scanned to determine estrous cyclicity. Pregnancy was evaluated after the first AI. Cure of metritis based on vaginal discharge <5 was greater for ampicillin than ceftiofur on d 5 (37.1 vs. 25.2%) and 7 (57.2 vs. 46.3%), but not on d 12 (82.0 vs. 85.0%). Cure of metritis based on vaginal discharge <5 and no fever was greater for ampicillin than for ceftiofur only on d 7 (50.4 vs. 37.9%), but not on d 5 (23.1 vs. 17.6%) and 12 (66.1 vs. 67.4%). Cows with puerperal metritis had reduced cure compared with cows with fetid discharge on d 5 (30.5 vs. 12.8%), 7 (55.2 vs. 33.6%), and 12 (72.0 vs. 61.1%). The proportion of cows with fever on any day after therapy started did not differ between treatments. Fifty-three percent of cows with metritis based on fetid discharge developed fever after initiating antimicrobial therapy. Cows receiving ampicillin had less prevalence of purulent vaginal discharge than those treated with ceftiofur (57.7 vs. 67.8%), but they were both greater than cows without metritis (21.9%). Prevalence of cytological endometritis did not differ between ampicillin and ceftiofur (30.0 vs. 25.4%), but they were both greater than cows without metritis (14.5%). The proportion of estrous cyclic cows (75.0%) and P/AI did not differ among treatments (ampicillin = 28.0% vs. ceftiofur = 28.3% vs. without metritis = 30.5%). Clinical cure was faster for ampicillin than for ceftiofur, but on study d 12 both treatments resulted in similar cure. Clinical cure was less for cows with puerperal metritis than for cows with fetid uterine discharge. Despite differences in uterine health, P/AI at the first insemination did not differ among treatments.     

Integration of statistical inferences and machine learning algorithms for prediction of metritis cure in dairy cows

The study's objectives were to identify cow-level and environmental factors associated with metritis cure to predict metritis cure using traditional statistics and machine learning algorithms. The data set used was from a previous study comparing the efficacy of different therapies and self-cure for metritis. Metritis was defined as fetid, watery, reddish-brownish discharge, with or without fever. Cure was defined as an absence of metritis signs 12 d after diagnosis. Cows were randomly allocated to receive a subcutaneous injection of 6.6 mg/kg of ceftiofur crystalline-free acid (Excede, Zoetis) at the day of diagnosis and 3 d later (n = 275); and no treatment at the time of metritis diagnosis (n = 275). The variables days in milk (DIM) at metritis diagnosis, treatment, season of the metritis diagnosis, month of metritis diagnostic, number of lactation, parity, calving score, dystocia, retained fetal membranes, body condition score at d 5 postpartum, vulvovaginal laceration score, the rectal temperature at the metritis diagnosis, fever at diagnosis, milk production from the day before to metritis diagnosis, and milk production slope up to 5, 7, and 9 DIM were offered to univariate logistic regression. Variables included in the multivariable logistic regression model were selected from the univariate analysis according to P-value. Variables were offered to the model to assess the association between these factors and metritis cure. Additionally, the univariate logistic regression variables were offered to a recursive feature elimination to find the optimal subset of features for a machine learning algorithms analysis. Cows without vulvovaginal laceration had 1.91 higher odds of curing of metritis than cows with vulvovaginal laceration. Cows that developed metritis at >7 DIM had 2.09 higher odds of being cured than cows that developed metritis at ≤7 DIM. For rectal temperature, each degree Celsius above 39.4°C led to lower odds to be cured than cows with rectal temperature ≤39.4°C. Furthermore, milk production slope and milk production difference from the day before to the metritis diagnosis were essential variables to predict metritis cure. Cows that had reduced milk production from the day before to the metritis diagnosis had lower odds to be cured than cows with moderate milk production increase. The results from the multivariable logistic regression and receiver operating characteristic analysis indicated that cows developing metritis at >7 DIM, with increase in milk production, and with a rectal temperature ≤39.40°C had increased likelihood of cure of metritis with an accuracy of 75%. The machine learning analysis showed that in addition to these variables, calving-related disorders, season, and month of metritis event were needed to predict whether the cow will cure or not from metritis with an accuracy ≥70% and F1 score (harmonic mean between precision and recall) ≥0.78. Although machine learning algorithms are acknowledged as powerful tools for predictive classification, the current study was unable to replicate its potential benefits. More research is needed to optimize predictive models of metritis cure.     

Susceptibility of Escherichia coli isolated from uteri of postpartum dairy cows to antibiotic and environmental bacteriophages. Part II: In vitro antimicrobial activity evaluation of a bacteriophage cocktail and several antibiotics

The use of pathogenic-specific antimicrobials, as proposed by bacteriophage therapy, is expected to reduce the incidence of resistance development. Eighty Escherichia coli isolated from uteri of Holstein dairy cows were phenotypically characterized for antimicrobial resistance to ampicillin, ceftiofur, chloramphenicol, florfenicol, spectinomycin, streptomycin, and tetracycline by broth microdilution method. The lytic activity of a bacteriophage cocktail against all isolates was performed by a similar method. Additionally, the effect of different concentrations of antimicrobials and multiplicities of infections (MOI) of the bacteriophage cocktail on E. coli growth curve was measured. Isolates exhibited resistance to ampicillin (33.7%), ceftiofur (1.2%), chloramphenicol (100%), and florfenicol (100%). All strains were resistant to at least 2 of the antimicrobial agents tested; multidrug resistance (≥3 of 7 antimicrobials tested) was observed in 35% of E. coli isolates. The major multidrug resistance profile was found for ampicillin-chloramphenicol-florfenicol, which was observed in more than 96.4% of the multidrug-resistant isolates. The bacteriophage cocktail preparation showed strong antimicrobial activity against multidrug-resistant E. coli. Multiplicity of infection as low as 10⁻⁴ affected the growth of the E. coli isolates. The ratio of 10 bacteriophage particles per bacterial cell (MOI = 10¹) was efficient in inhibiting at least 50% of all isolates. Higher MOI should be tested in future in vitro studies to establish ratios that completely inhibit bacterial growth during longer periods. All isolates resistant to florfenicol were resistant to chloramphenicol and, because florfenicol was recently introduced into veterinary clinics, this finding suggests that the selection pressure of chloramphenicol, as well as other antimicrobials, may still play a relevant role in the emergence and dissemination of florfenicol resistance in E. coli. The bacteriophage cocktail had a notable capacity to inhibit the in vitro growth of E. coli isolates, and it may be an attractive alternative to conventional treatment of metritis by reducing E. coli in uteri of postpartum dairy cows.     

Rectal temperature, calving-related factors, and the incidence of puerperal metritis in postpartum dairy cows

The objectives of this study were as follows: 1) to evaluate the association among abnormal calving, parity, and season on the incidence of puerperal metritis (PM) and clinical endometritis (CE) during d 3 to 13 and 20 to 30 postpartum, respectively; 2) to describe the rectal temperature (RT) of cows with PM before diagnosis; and 3) to document associations among PM, CE, and reproductive performance in lactating dairy cows. This study followed a prospective observational study design. Cows were classified as having an abnormal calving status (AC), i.e., cows calving with dystocia, twins, retained fetal membranes, or some combination of these conditions, and having a normal calving status (NC). Daily RT was recorded from d 3 to 13 postpartum for all cows, and health examinations were performed on cows that appeared not well. A total of 450 calvings were evaluated. Cows with an AC had greater odds of PM than cows with NC [adjusted odds ratio (AOR) = 4.8; 95% confidence interval (CI) = 2.9 to 8.0). A season by parity interaction showed that primiparous cows that calved during the warm season had lower AOR of PM than during the cool season (0.24; 95% CI = 0.09 to 0.62), whereas multiparous cows did not have seasonal effects on PM (1.43; 95% CI = 0.65 to 3.18). Cows with AC have greater AOR for CE than cows with NC (2.8; 95% CI = 1.7 to 4.9), and greater AOR of CE were detected in cows diagnosed with PM than in cows without PM (2.2; 95% CI = 1.1 to 3.9). Rectal temperature in cows with PM increased significantly 24 h before diagnosis of PM, reaching 39.2 ± 0.05°C on the day of diagnosis. In cows with PM and fever at diagnosis, the RT began to increase from 72 to 48 h before the diagnosis of PM and continued to increase to 39.7 ± 0.09°C on d 0 (day of diagnosis). Nonetheless, cows with PM without fever at diagnosis had no daily increases in RT before diagnosis of PM. Still, the RT on d 0 was different from cows without PM. Cows without PM had a stable RT (38.6 ± 0.01°C). There were no detected differences in first-service conception risk or cumulative pregnancy risk by 150 d postpartum between cows with or without PM. Still, a season effect on first-service conception AOR (warm vs. cool = 0.98; 95% CI = 0.18 to 0.72) and accumulated pregnancy AOR by 150 d postpartum was detected (warm vs. cool = 0.18; 95% CI = 0.10 to 0.33).     

Invited review: Antibiotic treatment of metritis in dairy cows: A systematic approach

Acute puerperal metritis (APM) is an acute systemic illness with fever ≥39.5°C and signs of toxemia due to an infection of the uterus occurring within 21 d after parturition. Because of the infectious nature of APM, antibiotics are considered beneficial for its treatment. Each use of an antimicrobial drug, however, is associated with selective pressure for the emergence of resistant bacteria. Hence, there is a significant need to encourage prudent use of antibiotics and alternative therapies to antibiotics. Therefore, the objective of this study was to systematically review the current literature on treatment of APM. A comprehensive and systematic literature search was conducted utilizing the PubMed and CAB Abstracts databases to identify literature focusing on the antibiotic therapy of puerperal metritis in the cow. After application of specific exclusion criteria, 21 publications comprising 23 trials remained for final evaluation. Data extraction revealed that the majority of the studies (n = 19) were attributable to the highest evidence level. Of 21 studies controlled, 11 had an untreated group and 3 a positive control group. The majority of the studies (n = 17) applied ceftiofur for the treatment of APM. Concerning the efficacy of ceftiofur, 7 studies observed clinical improvement, whereas none found improved reproductive performance. Fewer than half of the studies (n = 10) performed a bacteriological examination and only 4 implemented an antibiotic susceptibility test. Also, 3 studies (13.0%) described a self-cure rate per se. Little attention was given to the issue of bacterial resistance (n = 3), the need for reducing the application of antibiotics (n = 2), or guidelines for prudent use of antibiotics (n = 1). Our findings demonstrate that implementation of bacteriological examinations, sensitivity testing, and determination of minimum inhibitory concentrations, as well as reporting and discussion of critical issues (e.g., self-cure rates, resistance, prudent drug use), were suboptimal. On the other hand, the quality of studies on the treatment of APM was good, as indicated by evidence level 1. Nevertheless, more high-quality research considering self-cure rates is necessary to address critical issues related to APM and crucial to the dairy industry, such as resistance, prudent use of antibiotics, animal welfare, and cost-benefit ratios.     

Short communication: Assessment of biomarkers of inflammation in the vaginal discharge of postpartum dairy cows diagnosed with clinical metritis

Clinical metritis, characterized by the presence of an enlarged uterus and abnormal red-brownish foul-smelling vaginal discharge (VD), is a prevalent condition that causes important economic losses to dairy operations. The accurate diagnosis and treatment of this disease can help decrease its negative effects on the well-being and performance of dairy cows. The objectives of this study were to assess (1) the concentration of haptoglobin (Hp) and neutrophil-derived haptoglobin–matrix metalloproteinase 9 (Hp-MMP 9) in the VD of postpartum cows; and (2) the correlation between Hp and Hp-MMP 9 concentrations in serum and VD. Fifty-three dairy cows from 4 farms in central Pennsylvania were enrolled in this observational study.. Postpartum cows (7 ± 3 DIM) were screened using a Metricheck device to assess VD score (VDS): 1, clear fluid (n = 4); 2, < 50% white purulent fluid (n = 14); 3, > 50% white purulent fluid (n = 8); 4, red-brownish watery fluid without fetid smell (n = 22); and 5, fetid red-brownish watery fluid (n = 5). Blood and VD samples were collected for assessment of Hp and Hp-MMP 9 concentrations. Cows with a VDS of 4 or 5 (VDS4/5) had higher serum Hp concentrations than cows with a VDS of 1, 2, or 3 (VDS1/3; 93 ± 187 µg/mL vs. 59 ± 106 µg/mL, respectively). Similarly, cows with VDS4/5 had higher VD Hp concentrations than cows with VDS1/3 (73 ± 56 µg/mL vs. 17 ± 16 µg/mL, respectively). We found a significant correlation (0.37) between Hp levels in serum and in VD. We found no difference in serum Hp-MMP 9 between VDS4/5 and VDS1/3 cows. The VD concentrations of Hp-MMP 9 were higher in VDS4/5 cows than in VDS1/3 cows (7,629 ± 9,847 ng/mL vs. 1,567 ± 2,165 ng/mL, respectively). The correlation between Hp-MMP 9 in serum and VD was 0.22; nevertheless, it was not statistically significant. Interestingly, Hp and Hp-MMP 9 concentrations were higher in VD samples than in serum, regardless of VDS. Results from this study suggest that inflammatory biomarkers may be increased in cows with a VDS of 4 or 5. Further research should be aimed at elucidating the processes involved in inflammatory biomarker production and transportation in the uterus, as well as the effect of these biomarkers on endometrial cells.     

Evaluation of a systemic antibiotic treatment of toxic puerperal metritis in dairy cows

The objective of this study was to evaluate the efficacy and economic efficiency of a systemic treatment of toxic puerperal metritis in dairy cows with ceftiofur. Cows with abnormal vaginal discharge at a postpartum examination (d 4 to 6 after calving) and a rectal temperature > or = 39.5 degrees C were assigned to three treatment groups. Cows in group 1 (n = 70) received 600 mg of ceftiofur intramuscularly (i.m.) on 3 consecutive days. Cows in group 2 (n = 79) received an intrauterine treatment with antibiotic pills consisting of 2500 mg of ampicillin and 2500 mg of cloxacillin and an additional 6000 mg (i.m.) of ampicillin. This treatment was performed on 3 consecutive days. Cows in group 3 (n = 78) received the same intrauterine treatment as in group 2. In addition, 600 mg of ceftiofur was administered i.m. on 3 consecutive days. Body temperature was recorded daily for 6 d after first treatment. There were no significant differences among the groups regarding clinical efficacy at d 6 after first treatment. The cure rates based on rectal temperatures declining to below 39.5 degrees C on d 6 after treatment were 82.9, 84.8, and 84.6% for groups 1, 2, and 3, respectively. Reproductive performance did not differ significantly between group 1 and groups 2 and 3 for any of the measures tested. A financial analysis with 87 different cost scenarios demonstrated that a systemic treatment of toxic puerperal metritis in cattle with ceftiofur is an effective alternative to the combination of local and systemic treatments.     

Feed restriction to induce and meloxicam to mitigate potential systemic inflammation in dairy cows before calving

Most dairy cows experience a transient decrease in feed intake in the 1 to 2 wk before calving, which has been associated with systemic inflammation (SI), indicated by increased blood haptoglobin (Hp) concentration. We aimed to characterize the association between prepartum decrease in feed intake and the onset of SI and, if present, the ability of meloxicam (MEL), a non-steroidal anti-inflammatory drug, to mitigate SI. Holstein cows (n = 45) were assigned to control (n = 13), feed restriction (FR) untreated (FR-U; n = 15), and FR treated with MEL (FR-T; n = 17) groups. Daily feed intake was measured from ?22 d from expected parturition until 35 d postpartum. Control cows were fed ad libitum, whereas FR-U and FR-T cows were reduced to 60% of their average intake for 4 consecutive days (?15 to ?12 d from expected calving). The FR-T cows received MEL (0.5 mg/kg of body weight) once daily for 4 consecutive days (?13 to ?10 d from expected calving). Blood samples were collected ?22, ?15, ?14, ?13, ?12, ?10, ?7, ?5, ?3, 0, 1, 3, 5, 7, 15, 22, and 35 d relative to calving to measure serum concentrations of total calcium, total protein, albumin, globulin, cholesterol, urea, glucose, gamma-glutamyl transferase, aspartate aminotransferase, glutamate dehydrogenase, β-hydroxybutyrate, nonesterified fatty acids, Hp, and insulin-like growth factor-1. Serum concentrations of lipopolysaccharide-binding protein were measured ?22, ?15, ?14, ?13, ?12, and ?10 d from expected calving. Simplified glucose tolerance tests were performed on ?15, ?12, ?5, 1, and 5 d relative to calving. Mixed linear regression models were used to assess the effects of FR and MEL on each metabolite. The interaction between treatment group and blood sampling day was forced into each model. All models accounted for body condition score, parity, and the cow as a random effect. Nonesterified fatty acids concentrations in both the FR-U and FR-T groups significantly increased from the second until the last day of FR. Feed restriction increased urea concentrations compared with the control group on ?14 d but decreased urea concentrations on ?10 d from expected calving. Control cows had greater β-hydroxybutyrate concentrations compared with FR cows on 15, 21, and 35 d postpartum. For all other metabolites, no differences were found. This model of FR produced substantial fat mobilization but based on serum Hp and lipopolysaccharide-binding protein concentrations did not generate measurable SI; therefore, we were unable to evaluate the ability of MEL to mitigate SI.     

Prevalence,antimicrobial susceptibility,and molecular characterization of Staphylococcus aureus isolated from dairy herds in northern China

Staphylococcus aureus is one of the main pathogens involved in dairy cow mastitis. Monitoring of antibiotic use would prove useful to assess the risk of Staph. aureus in raw milk. The objective of this work was to investigate the prevalence of Staph. aureus strais isolated from raw milk in northern China, and to characterize antimicrobial susceptibility of these strains and their key virulence genes. In total, 195 raw milk samples were collected from 195 dairy farms located in 4 cities of northern China from May to September 2015. Out of 195 samples, 54 (27.7%) were positive for Staph. aureus. Among these 54 samples, 54 strains of Staph. aureus were isolated, and 16 strains were identified as methicillin-resistant Staph. aureus. The strains exhibited high percentages of resistance to penicillin G (85.2%), ampicillin (79.6%), and erythromycin (46.3%). Moreover, 72% of the strains showed resistance to more than one antimicrobial agent. Overall, 63% of penicillin-resistant strains possessed the blaZ gene, and 60% of the erythromycin-resistant strains possessed erm(A), erm(B), erm(C), msr(A), or msr(B) genes with 8 different gene patterns. All isolates resistant to gentamicin, kanamycin, and oxacillin carried the aac6'-aph2”, ant(4')-Ia, and mecA genes, respectively. Two tet(M)-positive isolates carried specific genes of the Tn916-Tn1545 transposon. The most predominant virulence genes were sec, sea, and pvl, which encode staphylococcal enterotoxins (sec and sea) and Panton-Valentine leukocidin, respectively. Thirty-two isolates (59.2%) harbored one or more virulence genes. The majority of Staph. aureus strains were multidrug resistant and carried multiple virulence genes, which may pose a risk to public health. Our data indicated that antimicrobial resistance of Staph. aureus was prevalent in dairy herds in northern China, and that antibiotics, especially penicillin G and ampicillin, to treat mastitis caused by Staph. aureus should be used with caution in northern China.     

Evaluation of effects of metritis management in a complex dairy herd health management program

Evaluating the effects of all interventions in a dairy herd, including the effects of various herd health management programs (HHMP), is highly relevant. A traditional randomized controlled trial is the gold standard but is likely practically impossible or prohibitively expensive to use for a general evaluation of a HHMP. Generalizability may also be poor because of the dynamics of the production contexts. In this study, we demonstrate an approach for evaluating the effects of an HHMP in the field, specifying an intervention theory for an ongoing HHMP in the context of the Danish dairy industry. As an example, we suggest one coherent analytical approach for studying the possible effects on milk production of systematic postpartum examinations of vaginal discharge, which is supposed to improve detection and treatment of metritis or endometritis. This routine is one component of the HHMP. The data consisted of 121 herds and 76,953 lactations over a 15-yr period. For parity group 1, the negative effects of metritis (despite treatment) on 305-d milk production after a normal calving were reduced by 116 kg of energy-corrected milk after enrollment in the HHMP. For parity group 2 and parity group >2, enrollment in the HHMP resulted in a 129-kg and an 80-kg energy-corrected milk yield increase in milk production, respectively. The results indicate that effects of the HHMP existed, which were mediated through improved metritis detection. This study demonstrates the importance of a clear-cut intervention theory, although even with a theory, the research question can be too herd and context specific. In such a case, a within-herd randomized controlled trial study design seems to be the only way to achieve a valid result for a given herd, and acquiring valid results from an observational multi-herd study will be very difficult.     

Differences in uterine and serum metabolome associated with metritis in dairy cows

Objectives were to evaluate differences in the uterine and serum metabolomes associated with metritis in dairy cows. Vaginal discharge was evaluated using a Metricheck device (Simcro) at 5, 7, and 11 d in milk (DIM; herd 1) or 4, 6, 8, 10, and 12 DIM (herd 2). Cows with reddish or brownish, watery, and fetid discharge were diagnosed with metritis (n = 24). Cows with metritis were paired with herdmates without metritis (i.e., clear mucous vaginal discharge or clear lochia with ≤50% of pus) based on DIM and parity (n = 24). Day of metritis diagnosis was considered study d 0. All cows diagnosed with metritis received antimicrobial therapy. The metabolome of uterine lavage collected on d 0 and 5, and serum samples collected on d 0 were evaluated using untargeted gas chromatography time-of-flight mass spectrometry. Normalized data were subjected to multivariate canonical analysis of population using the MultBiplotR and MixOmics packages in R Studio. Univariate analyses including t-test, principal component analyses, partial least squares discriminant analyses, and pathway analyses were conducted using Metaboanalyst. The uterine metabolome differed between cows with and without metritis on d 0. Differences in the uterine metabolome associated with metritis on d 0 were related to the metabolism of butanoate, amino acids (i.e., glycine, serine, threonine, alanine, aspartate, and glutamate), glycolysis and gluconeogenesis, and the tricarboxylic acid cycle. No differences in the serum metabolome were observed between cows diagnosed with metritis and counterparts without metritis on d 0. Similarly, no differences in uterine metabolome were observed between cows with metritis and counterparts not diagnosed with metritis on d 5. These results indicate that the establishment of metritis in dairy cows is associated with local disturbances in amino acid, lipid, and carbohydrate metabolism in the uterus. The lack of differences in the uterine metabolome on d 5 indicates that processes implicated with the disease are reestablished by d 5 after diagnosis and treatment.     

Minimum inhibitory concentrations of frequently used antibiotics against Escherichia coli and Trueperella pyogenes isolated from uteri of postpartum dairy cows

The objective of this study was to determine minimum inhibitory concentrations (MIC) of frequently used antimicrobials for Escherichia coli and Trueperella pyogenes isolated from postpartum bovine uteri of cows with acute puerperal metritis (APM, n = 67), cows suspected to have APM (n = 37), and healthy cows (n = 37) and to evaluate possible differences in MIC according to clinical signs. Cows with APM had reddish-brown, fetid vaginal discharge and rectal temperature (RT) ≥39.5°C within 21 d in milk; cows suspected to have APM had either reddish-brown, fetid vaginal discharge or RT ≥39.5°C within 21 d in milk; and healthy cows had neither fetid discharge nor RT ≥39.5°C. Samples were collected from cows on commercial dairy herds (n = 7) using the cytobrush technique. A total of 37 T. pyogenes isolates and 85 E. coli isolates were tested. Ceftiofur, a third-generation cephalosporin that is often used to treat APM, was the focus of analysis. Trueperella pyogenes and E. coli were isolated more often from samples of cows with APM (46 and 90%, respectively) compared with samples from healthy cows (19 and 54%, respectively). Regarding cows suspected to have APM, T. pyogenes and E. coli were numerically more often isolated (30 and 70%, respectively) than in healthy cows (19 and 54%, respectively). Minimum inhibitory concentrations of ceftiofur were low. For T. pyogenes and E. coli, MIC₅₀ (concentration that inhibited growth of 50% of isolates) were 0.25 and 0.5 µg/mL and MIC₉₀ (concentration that inhibited growth of 90% of isolates) were 0.5 and 1 µg/mL, respectively. Although ceftiofur inhibited all T. pyogenes at the highest concentration tested (64 µg/mL), the growth of 5.9% of E. coli was not impaired. Recently, ampicillin has been suggested as an alternative treatment for APM. Although the T. pyogenes isolates exhibited low MIC in general (MIC₅₀ ≤0.015 µg/mL and MIC₉₀ = 0.06 µg/mL) and 81.1% of all T. pyogenes could be inhibited at the lowest ampicillin concentration tested, 11.8% of the E. coli isolates were not impaired at the highest concentration (64 µg/mL) tested in this study. The MIC₅₀ and MIC₉₀ of E. coli were 4 and ≥128 µg/mL, respectively. We detected no difference in the MIC distributions of ceftiofur or ampicillin among isolates from the 3 APM groups. In summary, E. coli with high MIC against ceftiofur as well as against ampicillin were found in this study.     

Metritis diagnosis and treatment practices in 45 dairy farms in California

Minimizing the risk of antimicrobial-resistant organisms and antimicrobial residues in dairy and dairy beef products is a topic of nationwide interest. To design an effective outreach program on judicious use of antimicrobials, it is imperative to describe actual practices on dairies. Thus, our objective was to report current practices for metritis treatment in 45 dairies in California. Data were collected using a survey tool that included questions on systemic antimicrobial treatments, intrauterine treatments, supportive treatments, and treatment records. Researchers collected information based on cow-side observations and responses from fresh cow evaluators. Most dairies (n = 33; 70%) performed rectal exams for vaginal discharge (VD) evaluation and based systemic antimicrobial administration on the presence of (a) abnormal VD (fetid and nonfetid), regardless of fever (n = 11; 25%); (b) fever regardless of the odor of abnormal VD (n = 11; 25%); (c) fetid VD and fever (n = 1; 2%); (d) fetid VD (n = 3; 9%); or (e) fetid VD or fever (n = 3; 9%). Some dairies (n = 11; 24%) only detected abnormal VD if visible on tail, vulva, or floor; treatment decisions were based on the presence of abnormal VD (n = 9; 20%) or abnormal VD and fever (n = 2; 4%). On 3 (6%) dairies, VD characteristics were evaluated after rectal palpation, but no systemic treatments were administered. Cows exhibiting 1 nonspecific sign of health disorder (depressed general appearance, lack of appetite, or drop in milk yield) plus fever (n = 13; 29%), or even regardless of fever (n = 6; 13%), were treated with systemic antimicrobials. Overall, 8 (18%) dairies justified systemic antimicrobial treatment based on fever alone. Systemic antimicrobials used were ceftiofur products [ceftiofur hydrochloride (n = 26; 55%), ceftiofur sodium (n = 3; 7%), and ceftiofur crystalline-free acid (n = 8; 18%)], penicillin procaine (n = 8; 18%), and ampicillin (n = 3; 7%). Intrauterine infusions were done with penicillin procaine (n = 1; 2%) or oxytetracycline (n = 12; 27%). The dose and treatment length varied across dairies for i.m. ceftiofur hydrochloride (15 to 45 mL/d for 3 to 5 d), s.c. ceftiofur crystalline-free acid (20 to 25 mL once or twice), i.m. ceftiofur sodium (15 to 30 mL/d for 3 to 5 d), i.m. penicillin procaine (20 to 60 mL/d for 3 to 6 d), and i.m. ampicillin (25 to 35 mL/d for 3 to 6 d). The antimicrobial drug of choice, the dose, and the treatment length for metritis varied across the study dairies. Based on accepted industry best-management practices for metritis, a need exists to educate fresh cow evaluators on signs of health disorder indicative of metritis and on appropriate antimicrobial treatment regimens.     

Serum calcium dynamics within the first 3 days in milk and the associated risk of acute puerperal metritis

The objectives of this study were to evaluate postpartum serum calcium dynamics for different parity groups of dairy cows and to assess whether serum calcium concentration on d 0, 1, and 3 postpartum was associated with the risk of developing acute puerperal metritis (APM). The study took place on a commercial dairy farm in northern Germany and included 4,043 Holstein dairy cows. Calving difficulties, such as dystocia, twins, or stillbirth, were recorded. Blood samples were obtained on d 0, 1, and 3 after calving for analysis of serum calcium concentration. Animals were examined daily for clinical symptoms of retained placenta, APM, mastitis, and displaced abomasum until 10 d in milk. To determine serum calcium dynamics postpartum, we performed repeated-measures ANOVA with first-order autoregressive covariance. A logistic regression model was used to evaluate the association of serum calcium concentration with the risk of developing metritis. Serum calcium concentration was affected by time relative to calving, parity, and APM. Increasing parity negatively affected serum calcium concentration on d 0 and 1. Serum calcium concentration reached its lowest level on d 1 and 3 in multiparous and primiparous cows, respectively. The concentration increased from d 1 to 3 in multiparous cows and decreased from d 0 to 3 in primiparous cows. The association of APM and serum calcium dynamics varied by parity. On d 3, serum calcium concentration was significantly lower in animals with subsequent APM than in those without APM. The overall incidence of APM was 12.0% (primiparous cows, 20.4%; multiparous cows, 8.6%). An association existed between serum calcium concentration on d 3 after calving and APM. Primiparous cows had an odds ratio of 0.12 for serum calcium concentration on d 3, indicating that a primiparous cow with serum calcium concentration of 2.5 mmol/L had a 88% lower chance of developing APM compared with a cow with a concentration of 1.5 mmol/L. Multiparous cows had an odds ratio of 0.34 for serum calcium concentration on d 3, indicating that a multiparous cow with serum calcium concentration of 2.5 mmol/L had a 66% lower chance of developing APM compared with a cow with a concentration of 1.5 mmol/L. Primiparous cows with low serum calcium concentration had the highest predicted probability of developing APM. Our results reveal a dynamic in serum calcium concentration in the first 3 d in milk. Consequently, the day of sampling and the observed risk period for hypocalcemia are important when conducting epidemiological studies to evaluate associations between hypocalcemia and clinical diseases.     

Increased serum serotonin improves parturient calcium homeostasis in dairy cows

Hypocalcemia in dairy cows is caused by the sudden increase in calcium demand by the mammary gland for milk production at the onset of lactation. Serotonin (5-HT) is a key factor for calcium homeostasis, modulating calcium concentration in blood. Therefore, it is hypothesized that administration of 5-hydroxy-l-tryptophan (5-HTP), a 5-HT precursor, can increase 5-HT concentrations in blood and, in turn, induce an increase in blood calcium concentration. In this study, 20 Holstein dairy cows were randomly assigned to 2 experimental groups. Both groups received a daily i.v. infusion of 1 L of either 0.9% NaCl (C group; n = 10) or 0.9% NaCl containing 1 mg of 5-HTP/kg of BW (5-HTP group, n = 10). Infusions started d 10 before the estimated parturition and ceased the day of parturition, resulting in at least 4 d of infusion (8.37 ± 0.74 d of infusion). Until parturition, blood samples were collected every morning before the infusions, after parturition samples were taken daily until d 7, and a final sample was collected on d 30. Milk yield was recorded during this period. No differences between groups were observed for blood glucose, magnesium, and β-hydroxybutyrate. Cows receiving the 5-HTP infusion showed an increase in fatty acid concentrations from d ?3 to ?1 before parturition. Serum 5-HT concentrations were increased at d ?4 related to parturition until d 5 postpartum in the 5-HTP group compared with the C group. In addition, cows from the 5-HTP group had increased 5-HT concentrations in colostrum, but not in mature milk, on d 7 postpartum. Serum calcium concentrations decreased in both groups around parturition; however, calcium remained higher in the 5-HTP group than in controls, with a significant difference between groups on d 1 (1.62 ± 0.08 vs. 1.93 ± 0.09 mmol/L in control and 5-HTP groups, respectively) and d 2 (1.83 ± 0.06 vs. 2.07 ± 0.07 mmol/L in control and 5-HTP groups, respectively). Additionally, colostrum yield (first milking) was lower in the 5-HTP group compared with the C group, but without consequences on colostrum IgG concentrations. Milk yield did not differ between groups during the rest of the experiment. The study data were consistent with the concept that infusion of 5-HTP to dairy cows increases blood 5-HT concentrations, which in turn is a significant regulatory component in the chain of effectors that affect calcium status around parturition, hence the occurrence of clinical or subclinical hypocalcemia.     

Association of time to metritis diagnosis with circulating concentration of metabolites,minerals, and haptoglobin in Jersey cows

Our objective was to investigate the association of early metritis [EMET, diagnosed at <5 d in milk (DIM)] and late metritis (LMET, diagnosed at ≥5 DIM) with circulating concentrations of energy metabolites, minerals, and haptoglobin (Hp) throughout the first 14 d postpartum. A total of 379 purebred Jersey cows were enrolled in a prospective cohort study from a single herd in west Texas. Cows were examined for metritis using the Metricheck device (Simcro Ltd.) at 4, 7, and 10 DIM. Cows identified by farm employees as possible metritis cases were also evaluated for metritis. Blood samples were collected for analysis of concentrations of Ca, Mg, and glucose at DIM 1 through 5, 7, 10, and 14. Albumin, urea, fructosamine, free fatty acids (FFA), creatinine, and β-hydroxybutyrate (BHB) were analyzed at DIM 3, 5, 7, 10, and 14, and Hp at DIM 1 through 5 and 7. Data were analyzed using the MIXED and PHREG procedures of SAS (SAS Institute Inc.). A series of mixed general linear models accounting for repeated measures were fitted to the data. The independent variables metritis [no metritis (NMET), EMET, and LMET], DIM of analyte assessment, and parity were forced in all models. Multivariable Cox proportional hazard models were built to assess the risk of pregnancy and culling within 150 DIM. The overall metritis incidence was 26.9% (EMET = 49; LMET = 53; NMET = 277). Average concentrations of glucose, Mg, and urea were not associated with metritis. The associations of Ca, creatinine, BHB, and fructosamine with metritis were dependent on the DIM of analyte assessment. Cows categorized as EMET and LMET had, on average, lower albumin and fructosamine compared with NMET cows. Both EMET and LMET cows had, on average, greater BHB than NMET cows. A greater FFA concentration was only observed in cows diagnosed with EMET compared with NMET cows (EMET = 0.58, LMET = 0.52, NMET = 0.48 mmol/L). Additionally, circulating Hp concentration was greater for LMET and EMET compared with NMET cows, and EMET cows had greater Hp compared with LMET cows (EMET = 1.15; LMET = 1.00; NMET = 0.84). In conclusion, several blood biomarkers were temporally associated with early versus late metritis diagnosis in postpartum Jersey cows. No meaningful differences were observed in production, reproduction, or culling between EMET and LMET cows. These results suggest that cows with EMET undergo a more severe degree of inflammation and negative energy balance compared with NMET cows.