Intrauterine infusion of a pathogenic bacterial cocktail is associated with the development of clinical metritis in postpartum multiparous Holstein cows

Metritis is a uterine disorder common in dairy cattle caused by bacterial infection, with greater incidence in the early postpartum period. The disease causes delayed uterine involution, with a fetid, watery, red-brown discharge, with animals presenting different clinical signs including fever, dullness, inappetence, decreased milk yield, and dehydration. We developed an in vivo model of clinical metritis in Holstein multiparous cows using a pathogenic cocktail of Escherichia coli, Fusobacterium necrophorum, and Trueperella pyogenes. A total of 36 multiparous cows were randomly allocated to 1 of 3 treatment groups of 12 animals each. Cows assigned to the control group received an intrauterine administration of sterile saline solution, those in the low-dose group received a bacterial inoculum containing 10⁶ cfu of Escherichia coli, Trueperella pyogenes, and Fusobacterium necrophorum; and those in the high-dose group received 10⁹ cfu of these same organisms, all within 24 h of parturition. Clinical signs; milk yield; dry matter intake; serum concentration of acute phase proteins, metabolites, and cytokines; and bacterial counts in vaginal discharge were measured from parturition until 14 d in milk. Based on clinical signs, cows in the low-dose group had a greater incidence of metritis (83.3%) compared with the control (9%) or high-dose (25%) group. The low-dose group also had lesser dry matter intake compared with controls and an acute phase protein profile that typified metritis. The low-dose group presented greater relative abundance of bacteria from the genus Fusobacterium in the vaginal discharge compared with control cows, and the high-dose group was similar to the control group. Unexpectedly, intrauterine infusion of cows with a lower dose of bacteria was more effective than a higher dose, indicating that the bacterial load may affect metritis incidence in a nonlinear manner. These findings demonstrate that clinical metritis can be experimentally induced in postpartum multiparous Holstein cows, creating a relevant model for the study of uterine diseases.     

Short communication: Diagnostic performance of on-farm bacteriological culture systems for identification of uterine Escherichia coli in postpartum dairy cows

The objectives of this study were to validate the performance of on-farm bacteriological culture systems for identification of Escherichia coli in the uterus of early postpartum dairy cows and to determine if an association is present between the results and the subsequent occurrence of puerperal metritis (PM). A prospective cohort study was conducted in one commercial Holstein dairy herd in which 400 cows were sampled between 24 and 48 h after parturition. Three bacteriological samples were obtained from the uterus of each cow and were cultured for identification of E. coli. One sample was cultured in a commercial bacteriology laboratory according to standard procedures for identification of E. coli and was considered as the reference test. The two other samples were cultured on the farm using the Tri-Plate (University of Minnesota, St. Paul, MN) and Petrifilm systems, and plate readings were done after 24 and 48 h of incubation (variables: Tri24h, Tri48h, Petri24h, Petri48h). Participating cows were followed until 21 days in milk to diagnose PM. The prevalence of PM and E. coli (from the reference test) in the cow population was 15.0 and 33.5%, respectively. Both on-farm culture systems were accurate compared with the reference test. The sensitivity and specificity were 97 and 100%, 99 and 100%, 100 and 92%, and 100 and 89% for Tri24h, Tri48h, Petri24h, and Petri48h, respectively. On-farm results for Tri24h, Tri48h, Petri24h, and Petri48h were associated with subsequent occurrence of PM. The results from this study support the use of the Tri-Plate and Petrifilm culture systems on dairy farms to identify the presence of E. coli in the uterus of postpartum cows.     

Evaluation of peripartal calcium status,energetic profile,and neutrophil function in dairy cows at low or high risk of developing uterine disease

In this prospective cohort study, Holstein cows considered to be at high risk of developing metritis (dystocia, twins, stillbirth, retained placenta, or their combination) were matched with herdmates at low risk of developing metritis (normal calving) and monitored daily for rectal temperature and uterine discharge during the first 12 d in milk (DIM). Blood was sampled on d 0, 1, and 3 postpartum for assessment of neutrophil phagocytic and oxidative burst activities. Blood was also sampled at 0, 1, 2, 3, 4, 7, and 12 DIM for determination of serum concentrations of Ca, K, Mg, nonesterified fatty acids, β-hydroxybutyrate, and glucose. On the basis of receiver operator characteristic curves, subclinical hypocalcemia (SCH) was defined as a serum Ca concentration ≤8.59 mg/dL in at least 1 sample in the first 3 DIM. The overall incidences of metritis and puerperal metritis were 47.3 and 30%, respectively. Concentration in blood and percentages of neutrophils undergoing phagocytosis and oxidative burst were all reduced in cows with SCH compared with normocalcemic cows. Cows with SCH were at a greater risk of developing fever, metritis, and puerperal metritis compared with normocalcemic cows. Among cows at low risk of developing metritis, those with SCH had a greater incidence of metritis (40.7%) compared with normocalcemic cows (14.3%). Similarly, among cows at high risk of developing metritis, cows with SCH had a greater incidence of metritis (77.8%) compared with normocalcemic cows (20.0%). Cows with SCH had elevated concentrations of nonesterified fatty acids and β-hydroxybutyrate compared with normocalcemic cows. The relative risk of developing metritis decreased by 22% for every 1 mg/dL increase in serum Ca. Neither metritis nor SCH influenced the resumption of estrous cyclicity by 38 DIM, but cows with SCH had a reduced pregnancy rate and a longer interval to pregnancy compared with normocalcemic cows. Finally, the population risk to develop uterine diseases attributable to SCH was 66.6% for metritis and 91.3% for puerperal metritis in the present study.     

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.     

Metagenomic analysis of the uterine bacterial microbiota in healthy and metritic postpartum dairy cows

At present, many bacterial species are validly known as etiological agents of dairy cattle metritis, yet the vast uncultured fraction has received no attention so far. The purpose of this study was to use culture-independent methods to describe and compare the uterine bacterial composition in healthy and metritic postpartum Holstein dairy cows. Both group-specific 16S ribosomal DNA PCR-denaturing gradient gel electrophoresis (DGGE) and clone library sequencing of broad-range 16S ribosomal DNA PCR revealed differences in the bacterial communities comparing healthy and metritic cows. Bacterial diversity in healthy and metritic uteri was greater and more complex than described previously by traditional culture methods. Sequences were assigned to 5 major groups (Gammaproteobacteria, Firmicutes, Fusobacteria, Bacteroidetes, and Tenericutes) and to uncultured bacteria. Additionally, DGGE suggested the presence of Actinobacteria. Most clone sequences in the metritic status libraries were affiliated with the phylum Fusobacteria. Many components, especially from other phyla, have not previously been isolated from cases of metritis. In the clone libraries from the healthy status dairy cows, Gammaproteobacteria was the most prominent group and most sequences showed high identity with Mannheimia varigena, Pasteurella hemolytica, and members of the phylum Tenericutes. Our data showed that the uterine bacterial community in postpartum dairy cows differed considerably between healthy and metritic cows and described the occurrence of a previously unrecognized extent of this diversity in the bovine intrauterine microbiota.     

Association between uterine disease and indicators of neutrophil and systemic energy status in lactating Holstein cows

The objective of this study was to evaluate the association between uterine disease and indicators of neutrophil (PMN) and systemic energy status in dairy cows. Peripheral blood (120 mL) was collected weekly from 84 Holstein cows for PMN isolation and plasma collection from calving until 42 d in milk (DIM). The final analysis included 80 cows. Of those, 20 cows were classified as having metritis (fetid uterine discharge and fever), 15 as having subclinical endometritis (SCE; ≥10% PMN on uterine cytology), and 45 as healthy controls. Plasma haptoglobin concentration was increased only in cows that developed metritis. Neutrophil glycogen content was reduced in cows developing metritis compared with healthy cows on the day of calving and at 7 and 42 DIM. Cows with SCE had lower PMN glycogen content than healthy cows at 7, 28, and 42 DIM. Blood glucose was affected by disease status within parity. Primiparous metritis cows had greater blood glucose concentrations than healthy primiparous cows. Multiparous metritis cows tended to have lower blood glucose concentration than multiparous SCE cows. Cows that developed metritis and SCE had or tended to have greater NEFA and BHBA than healthy cows, mainly around calving. At calving, cows that developed metritis had higher plasma estradiol concentration than healthy cows and greater plasma cortisol than cows that had SCE. Plasma insulin was not affected. Plasma glucagon was increased for SCE cows. Cows that developed uterine disease experienced a greater degree of negative energy balance and had decreased lower intracellular PMN glycogen levels, which could be a major predisposing factor for disease because of decreased availability of oxidative fuels.     

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.     

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.     

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.     

Inducing ovulation early postpartum influences uterine health and fertility in dairy cows

The objective of the current study was to evaluate the effect of GnRH early postpartum on induction of ovulation, uterine health, and fertility in dairy cows. Holstein cows without a corpus luteum (CL) at 17 ± 3 DIM were assigned randomly to receive i.m. GnRH (n = 245) at 17 ± 3 and 20 ± 3 DIM or remain as controls (n = 245). Ovaries were scanned by ultrasonography twice weekly totaling 4 examinations. Ovulation was characterized by the appearance of a CL ≥20 mm at any ultrasound or CL <20 mm in 2 consecutive examinations. Clinical and cytological endometritis were diagnosed at 35 DIM. Compared with control, GnRH increased ovulation up to 3.5 d after the last treatment (78.7 vs. 45.0%) and did not affect the prevalence of clinical endometritis (23.9 vs. 18.6%) or cytological endometritis (30.9 vs. 32.8%). Prevalence of clinical endometritis increased in cows that had calving problems (32.6 vs. 15.9%) and metritis (40.6 vs. 15.8%). Metritis increased prevalence of cytological endometritis (50.7 vs. 23.5%). Treatment with GnRH did not affect pregnancy per artificial insemination at 32 (37.6 vs. 38.6%) or 74 d after artificial insemination (35.0 vs. 31.5%), but reduced pregnancy loss (6.8 vs. 18.1%). No overall effect of GnRH treatment on hazard of pregnancy was observed; however, an interaction between GnRH treatment and ovulation showed that GnRH-treated cows that ovulated had increased hazard of pregnancy by 300 DIM compared with GnRH-treated and control cows that did not ovulate (hazard ratio = 2.0 and 1.3, respectively), but similar to control cows that ovulated (hazard ratio = 1.1). Gonadotropin-releasing hormone early postpartum induced ovulation without affecting uterine health, but failed to improve pregnancy per artificial insemination or time to pregnancy, although it reduced pregnancy loss.     

Relationship between bacteriological findings in the second and fourth weeks postpartum and uterine infection in dairy cows considering bacteriological results

The uterine lumen in early postpartum dairy cows is contaminated with different bacteria. The most relevant uterine pathogens are Escherichia coli and Trueperella pyogenes. Prevalence of α-hemolytic streptococci and coagulase-negative staphylococci (CNS) is also high; however, these pathogens are considered opportunistic. The overall objective of this study was to investigate effects of the intrauterine presence of E. coli, T. pyogenes, α-hemolytic streptococci, or CNS at 10 ± 1 d in milk (DIM) on the type of bacteria 2 wk later and their influence on uterine infections and subsequent reproductive performance. Furthermore, we set out to quantify 2 relevant methodological factors (i.e., laboratory and sampling instrument). Bacteriological samples were collected at 10 ± 1 and 24 ± 1 DIM from the uterine lumen using a cytobrush (CB). Vaginal mucus was classified by vaginoscopy. In a subsample, bacteriological results of 3 different laboratories and of CB and cotton swabs (CS) were compared. Samples of uterine discharge were collected at 10 ± 1 DIM and bacteriological samples were taken using CB and CS. Bacteria were identified and bacterial growth quantified on a 4-point scale. Animals infected with E. coli or T. pyogenes at 10 ± 1 DIM had a higher risk for an infection with the same bacterial species at 24 ± 1 DIM [E. coli relative risk (RR) = 3.7 and T. pyogenes RR = 2.9]. Moreover, the risk of being diagnosed with abnormal vaginal discharge at 24 ± 1 DIM increased in cows with E. coli (RR = 1.7) or T. pyogenes (RR = 1.7) at 10 ± 1 DIM. Uterine infection with α-hemolytic streptococci or CNS did not increase the risk of an infection with T. pyogenes or E. coli or abnormal vaginal discharge 2 wk later. Cows with E. coli at 10 ± 1 DIM or T. pyogenes at 24 ± 1 DIM had greater days to first artificial insemination than cows positive for the 3 remaining bacterial species. Cows with T. pyogenes at 10 ± 1 DIM had more days to pregnancy and more cows were culled when positive for E. coli at 10 ± 1 DIM. Agreement of bacteriological results of 3 different laboratories were significant for laboratory A + B and A + C for the CB and CS. The highest agreement considering the bacterial species was for E. coli. All results for laboratory A + B and A + C for the CB and CS were significant. The results generated from samples collected with CB agreed nicely with those from CS from each laboratory (laboratory A: 250/272; laboratory B: 264/272; laboratory C: 253/272).     

Risk factors associated with postpartum subclinical hypocalcemia in dairy cows

Our study objectives were to evaluate the association of prepartum plasma Mg concentrations with subclinical hypocalcemia (SCH) classification at parturition and to evaluate the association of other cow-level risk factors with SCH classification at calving or at 2 d in milk (DIM). A total of 301 animals from 2 dairy herds located in New York were enrolled in a cohort study. Blood samples were collected at approximately 1 wk before the expected calving date, within 4 h of calving, and at 2 DIM. Prepartum samples had plasma macromineral concentrations (Ca, K, Mg, P), albumin, and β-hydroxybutyrate analyzed. Samples collected at calving were analyzed for Ca only, and samples from 2 DIM had macromineral and albumin concentrations determined. Postpartum SCH was defined as Ca concentrations ≤2.1 mmol/L. The prevalence of SCH at calving was 2, 40, and 66% for first, second, and third or greater parities, respectively. Only 4% of cows could be classified with prepartum subclinical hypomagnesemia (Mg concentrations <0.8 mmol/L), which did not provide enough power to appropriately determine the association of plasma Mg with postpartum Ca concentrations and its effect on SCH classification. Multiparous cows with Ca concentrations ≤2.4 mmol/L in the prepartum period and third or greater parity cows had a higher risk of being categorized as SCH at calving [relative risk (RR) = 1.4 and 1.7, respectively]. The risk of SCH at 2 DIM was associated with the interaction of Ca status at calving and lameness score. Nonlame cows with Ca concentrations ≤2.1 mmol/L (RR = 3.2) and normocalcemic lame cows at parturition (RR = 3.4) were more likely to be SCH at 2 DIM compared with nonlame normocalcemic cows. In conclusion, we identified a prepartum Ca cut-point for identification of cows that are more likely to be classified as SCH at calving. Different risk factors were associated with SCH depending on the timing of diagnosis relative to parturition.     

Dynamics of the microbiota found in the vaginas of dairy cows during the transition period: Associations with uterine diseases and reproductive outcome

We investigated the microbiota found in the vaginas of Holstein dairy cows during the transition period and described the differences in bacterial composition and total bacterial load (TBL) associated with disease and fertility. Vaginal swabs were collected at ?7, 0, 3, and 7 d relative to parturition from 111 dairy cows housed on a commercial dairy farm near Ithaca, New York. Microbiota were characterized by next-generation DNA sequencing of the bacterial 16S rRNA gene, and TBL was determined by real-time quantitative PCR. We applied repeated-measures ANOVA to evaluate the associations of uterine disease and related risk factors with the microbiota and TBL. We estimated phylum-specific bacterial load by multiplying the TBL by the relative abundance of each phylum observed in the metagenomics results. We confirmed the validity of this approach for estimating bacterial load by enumerating the number of bacteria in an artificial sample mixed in vitro and in clinical and healthy vaginal samples. Phyla associated with uterine disease and related risk factors were Proteobacteria, Fusobacteria, and Bacteroidetes. Cows with retained placenta and healthy cows had similar TBL at the day of parturition, but at d 7 postpartum, cows with retained placenta showed a significantly higher TBL, mainly driven by higher estimated loads of Fusobacteria and Bacteroidetes. Cows diagnosed with metritis had a significantly higher estimated load of Proteobacteria at d ?7 and at calving and higher estimated loads of Fusobacteria in the postpartum samples. Additionally, the estimated load of Bacteroidetes at d 7 postpartum was higher for cows diagnosed with endometritis at 35 days in milk. Higher estimated loads of Fusobacteria and Bacteroidetes were also evident in cows with postpartum fever, in primiparous cows, in cows with assisted parturition, and in cows that gave birth to twins. Our findings demonstrated that microbiota composition and TBL were associated with known periparturient risk factors of uterine diseases and reproductive failure, including parity, assisted parturition, and retained fetal membranes.     

Symposium review: The implications of spontaneous versus synchronized ovulations on the reproductive performance of lactating dairy cows

Lactating dairy cows are classified as spontaneous ovulators, in which establishment of pregnancy depends on the accuracy of detection of behavioral estrus for correct timing of artificial insemination (AI). Development of the Ovsynch protocol, a hormonal protocol that synchronizes ovarian function, thereby allowing for timed AI (TAI) without the need to detect estrus, provided a management tool for increasing AI service rates but not pregnancies per AI (P/AI). A review of 7 randomized, controlled experiments that compared P/AI of cows inseminated after a detected estrus to that of cows receiving TAI after submission to Presynch-Ovsynch or Double-Ovsynch protocols supports that the newest programs for TAI yield more P/AI than cows inseminated after a detected estrus. The physiologic and endocrine mechanisms that explain how fertility programs increase P/AI are a culmination of over 20 yr of research aimed at increasing reproductive performance in lactating dairy cows. We illustrate the dramatic change in reproductive performance of US dairy cows over time by comparing the phenotypic trend in days open with the genetic trend in daughter pregnancy rate and the phenotypic trend in cow conception rate. Whereas days open increased from 1955 to 2000, days open from 2000 to 2010 dramatically decreased without a concurrent increase in the genetic trend for daughter pregnancy rate. By contrast, the dramatic decrease in days open over the past 20 yr is associated with a dramatic increase in the phenotypic trend in cow conception rate. Although many management factors affect P/AI, adoption and implementation of TAI programs that directly increase P/AI is an important component of the dramatic increase in reproductive performance in lactating dairy cows in the United States over the past 20 yr.     

Effects of a single transdermal administration of flunixin meglumine in early postpartum Holstein Friesian dairy cows: Part 1. Inflammatory and metabolic markers,uterine health,and indicators of pain

The objectives of this study were to assess the effects of a single transdermal administration of flunixin meglumine (FM) in early postpartum Holstein Friesian dairy cows on serum concentrations of inflammatory and metabolic markers, uterine health, and indicators of pain. The hypothesis was that the anti-inflammatory, antipyretic, and analgetic effects of the pharmaceutic agent would reduce systemic inflammation, resulting in improved metabolic and inflammatory profile, diminished incidence of metritis, and reduced expression of pain. A total of 500 cows (153 primiparous, 347 multiparous) from 3 different commercial dairy farms in the northeast of Germany were included in a randomized controlled clinical trial. Farms were preselected based on high haptoglobin concentrations in their fresh lactating cows. Cows were excluded if they had experienced dystocia, stillbirth, or twin birth, or if they showed any signs of milk fever, retained fetal membranes, or fever (>40°C). The cows were treated once with either FM (3.33 mg/kg) or a placebo as control (CON) through transdermal administration between 24 to 36 h postpartum (d 2). General health examinations were performed (daily from d 2–8 and additionally on d 15 postpartum), vaginal discharge was assessed using the Metricheck device (d 8 and 15 postpartum) and serum samples were analyzed for inflammatory and metabolic markers (d 2, 4, and 6 postpartum). Effects of treatment, parity, sampling day, and their interactions were evaluated using mixed effects models. Primiparous cows treated with FM showed lower serum haptoglobin concentrations (0.90 ± 0.08 vs. 1.17 ± 0.07 g/L; ± standard error of the mean) and higher serum albumin concentrations (35.5 ± 0.31 vs. 34.8 ± 0.31 g/L) on d 6 postpartum. They also had a lower risk for purulent vaginal discharge with or without a fever compared with CON cows on d 15 postpartum (odds ratio for CON vs. FM: 1.63, 95% CI: 1.26–2.00), and body temperature was lower throughout the first 15 d in milk (39.1 ± 0.11 vs. 39.2 ± 0.11°C). Multiparous cows treated with FM had lower serum β-hydroxybutyrate concentrations on d 4 postpartum (0.71 ± 0.05 vs. 0.78 ± 0.05 mmol/L) and d 6 postpartum (0.74 ± 0.05 vs. 0.80 ± 0.05 mmol/L). Regardless of parity, FM-treated cows were significantly less likely to abduct their tail from their body (14.3 vs. 23.6%) and show an arched back (27.9 vs. 39.7%) on the day after treatment compared with CON cows. It can be concluded that FM treatment slightly reduced inflammation and diminished the risk for metritis in primiparous cows, improved metabolic profile in multiparous cows, and reduced expressions of pain in all cows.     

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.     

Risk factors associated with colostrum quality in Norwegian dairy cows

The objectives of the present study were to evaluate colostrum quality in Norwegian dairy cows based on IgG content, and to identify associations between possible risk factors and low colostral IgG. A longitudinal cross-sectional survey on calf health in Norway was performed between June 2004 and December 2006. The participating dairy herds were randomly selected among herds registered in the Norwegian Dairy Herd Recording System as having at least 15 cow years. The participating farmers were requested to sample 10 mL of colostrum from the first milking after calving from 12 cows that had calved during the defined project period of 365 d. Colostrum samples from 1,250 cows from 119 herds were collected. The material consisted of 451, 337, 213, and 249 samples collected from cows in their first, second, third, and fourth parity or more, respectively. Analysis was performed on IgG content by using single radial immunodiffusion. Mixed models with herd as a cluster were fit by using grams of IgG per liter of colostrum as the dependent variable for the statistical analyses. The IgG content in the colostrum sampled ranged from 4 to 235 g/L, with a median of 45.0 g of IgG/L, with the 10th, 25th, 75th, and 90th percentiles being 23.1, 31.4, 63.6, and 91.6 g of IgG/L, respectively. Altogether, 57.8% of the samples contained less than the desired 50 g of IgG/L of colostrum. Cows in their fourth parity or more were found to have significantly higher levels of IgG per liter of colostrum than cows in their first or second parity. Colostrum from cows in their second parity had the lowest level of IgG. Cows calving during the winter months (December, January, and February) produced colostrum with a significantly lower IgG content compared with cows calving in any other season of the year. Somatic cell count, measured after calving, was significantly higher in cows producing colostrum of inferior quality compared with those producing high-quality colostrum. Of the total variation in colostrum quality, 13.7% could be explained by cluster effects within herd. The variation in IgG content in colostrum produced by Norwegian dairy cows indicates a need for improved colostrum quality control and subsequent adjustment of the colostrum feeding regimen to ensure a protective immunological status for newborn calves.     

Effects of urea infusion on the uterine luminal environment of dairy cows

Previous research indicates that high plasma urea nitrogen (PUN) concentrations are associated with decreased fertility in lactating dairy cows. The objective of this study was to monitor changes in the uterine environment during acute elevation of PUN. Lactating dairy cows (n = 8) were infused with saline or urea (0.01 g of urea/h per kg of body weight) through jugular vein catheters on d 7 after estrus. After 24 h, cows were switched to the opposite treatment for a second 24-h infusion period. Blood samples were collected every 2 h, and the pH within the lumen of the uterine horn ipsilateral to the corpus luteum was recorded every 6 h. At the end of each 24-h infusion period, 30 mL of sterile saline was flushed into the uterine lumen and immediately retrieved. Mean PUN concentration increased from 16.6 +/- 1.3 mg/dL during saline infusion to 22.6 +/- 1.3 mg/dL during urea infusion. Uterine pH decreased during urea infusion from 7.08 +/- 0.07 at 6 h to 6.88 +/- 0.08 at 18 h, but was unchanged during saline infusion (7.01 +/- 0.08 at 6 h to 7.06 +/- 0.07 at 18 h). Protein concentration, PGF(2alpha), and prostaglandin E(2) concentrations in uterine lavage samples were not different between treatments. The results of this study indicate that a short-term increase in PUN can exert direct effects on the uterine environment by decreasing uterine pH.     

Bacterial flora associated with udder cleft dermatitis in Dutch dairy cows

Udder cleft dermatitis (UCD) or udder sores is a skin lesion, characteristically located around the anterior junction between the udder and abdomen of dairy cows. It is a worldwide problem in dairy herds with a large effect on animal welfare. The etiology and possible infectious origins of UCD are largely unknown; however, specific bacterial or parasitic causes are suggested in the literature. Therefore the aim of this study was to investigate the possible bacteriological, mycological, or parasitic involvement in clinically scored UCD lesions. Bacteriological culture was performed on subcutaneous tissue samples taken postmortem at a depth of 5 to 10 mm of 87 mild or severe UCD lesions or from the same place in healthy cows. Fungal culture was performed on a subset of 22 subcutaneous tissue samples of severe UCD postmortem. To investigate the superficial flora, swabs were taken from normal skin or skin lesions of 15 live animals equally divided over 3 groups: healthy skin or mild and severe UCD lesions. Histopathology, to describe and classify the lesions and to assess the presence of mites, fungi, or bacteria, was performed on 128 tissue samples, taken separately. In severe UCD lesions, Trueperella pyogenes and Bacteroides pyogenes were more frequently present in deep tissue layers and in superficial layers, compared with the same layers in mild UCD lesions or healthy skin. Culturing and histopathology indicated no sign of involvement of treponemes, Staphylococcus aureus, Escherichia coli, fungi, or mites in the UCD lesions. Histopathological examination showed that the majority of the lesions were characterized by chronic aspecific inflammation. Severe UCD lesions more frequently showed chronic active inflammation on histopathology, compared with mild UCD lesions. Due to the cross-sectional character of this study, it is difficult to differentiate cause and effect; however, future preventive and curative measures against UCD should take into account the chronic and anaerobic nature of this illness.