Prolonged hyperinsulinemia increases the production of inflammatory cytokines in equine digital lamellae but not in striated muscle.

Hyperinsulinemia is the key feature of equine metabolic syndrome (EMS) which leads to debilitating sequelae. Hyperinsulinemia-associated laminitis (HAL) is one of the major sequelae of EMS, although the pathophysiological mechanisms are not well elucidated. Using an equine model, we hypothesized that expression of inflammatory markers would be increased in digital lamellae and striated muscle following prolonged hyperinsulinemia. Healthy Standardbred horses (5.4 ± 1.9 years) were alternately assigned to a prolonged euglycemic-hyperinsulinemic clamp (pEHC) or control group (n = 4 per group). Following a 48 h pEHC or a 48 h infusion of a balanced electrolyte solution (controls), biopsies were collected from digital lamellar tissue, skeletal muscle and cardiac muscle were obtained. All hyperinsulinemic horses developed laminitis regardless of previous health status at enrollment. Protein expression was quantified via Western blotting. A significant (P < 0.05) upregulation of the protein expression of heat shock protein 90 (HSP90), alpha 2 macroglobulin (A2M) and fibrinogen (α, ß isoforms), as well as inflammatory cytokines including interleukin-1ß were detected in digital lamellae following prolonged hyperinsulinemia. In contrast, protein expression of cytokines and acute phase proteins in heart and skeletal muscle was unchanged following hyperinsulinemia. Upregulation of inflammatory cytokines and acute phase proteins in digital lamellae during prolonged hyperinsulinemia may reveal potential biomarkers and novel therapeutic targets for equine endocrinopathic laminitis. Further, the lack of increase of inflammatory proteins and acute phase proteins in striated muscle following prolonged hyperinsulinemia may highlight potential anti-inflammatory and cardioprotective mechanisms in these insulin-sensitive tissues.

Influence of epidemiological and clinical factors in the reactogenicity to Comirnaty® vaccine in health care workers of a Spanish university teaching hospital (COVIVAC study).

OBJECTIVE: Comirnaty® is an mRNA vaccine against COVID-19 which has been administered to millions of people since the end of 2020. Our aim was to study epidemiological and clinical factors influencing reactogenicity and functional limitation after the first two doses of the vaccine in health care workers (HCWs). METHODS: Prospective post-authorization cohort study to monitor safety and effectiveness of the vaccine. RESULTS: Local side effects were mild and presented both with first and second dose of Comirnaty. Systemic side effects were more frequent after 2nd dose. Nevertheless, previous SARS-CoV-2 infection was associated with systemic effects after the first dose of the vaccine (OR ranging from 2 to 6). No severe adverse effects were reported. According to multivariate analysis, the degree of self-reported functional limitation after the first dose increased with age, female sex, previous COVID-19 contact, previous SARS-CoV-2 infection, and Charlson Comorbidity Index (CCI). After the second dose, the degree of functional limitation observed was lower in those with previous SARS-CoV-2 infection, and it was positively associated to the degree of functional limitation after the first dose. CONCLUSIONS: Systemic adverse effects were more frequent after the second dose of Comirnaty. Previous SARS-CoV-2 infection was associated with systemic effects after the first dose. Age, female sex, previous COVID-19, previous isolation due to COVID-19 contact, and CCI showed to be independent predictors of the degree of functional limitation after the 1st dose of Comirnaty®. After the 2nd dose, the degree of functional limitation was lower in those who previously had SARS-CoV-2 infection.

Prehospital treatment with zalunfiban (RUC-4) in patients with ST- elevation myocardial infarction undergoing primary percutaneous coronary intervention: Rationale and design of the CELEBRATE trial.

BACKGROUND: Early and complete restoration of target vessel patency in ST-elevation myocardial infarction (STEMI) is associated with improved outcomes. Oral P2Y12 inhibitors have failed to demonstrate either improved patency or reduced mortality when administered in the prehospital setting. Thus, there is a need for antiplatelet agents that achieve prompt and potent platelet inhibition, and that restore patency in the prehospital setting. Zalunfiban, a novel subcutaneously administered glycoprotein IIb/IIIa inhibitor designed for prehospital administration, has shown to achieve rapid, high-grade platelet inhibition that exceeds that of P2Y12 inhibitors. Whether prehospital administration of zalunfiban can improve clinical outcome is unknown. HYPOTHESIS: The present study is designed to assess the hypothesis that a single, prehospital injection of zalunfiban given in the ambulance, in addition to standard-of-care in patients with STEMI with intent to undergo primary percutaneous coronary intervention (PCI) will improve clinical outcome compared to standard-of-care with placebo. STUDY DESIGN: The ongoing CELEBRATE trial (NCT04825743) is a phase 3, randomized, double-blinded, placebo-controlled, international trial. Patients with STEMI intended to undergo primary PCI will receive treatment with a single subcutaneous injection containing either zalunfiban dose 1 (0.110 mg/kg), zalunfiban dose 2 (0.130 mg/kg) or placebo, and the study drug will be administered in the ambulance before transportation to the hospital. A target of 2499 patients will be randomly assigned to one of the treatment groups in a 1:1:1 ratio, ie, to have approximately 833 evaluable patients per group. The primary efficacy outcome is a ranked 7-point scale on clinical outcomes. The primary safety outcome is severe or life-threatening bleeding according to the Global Use of Strategies to Open Occluded Coronary Arteries (GUSTO) criteria. SUMMARY: The CELEBRATE trial will assess whether a single prehospital subcutaneous injection of zalunfiban in addition to standard-of-care in patients with STEMI with intent to undergo primary PCI will result in improved clinical outcome.

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.

Effects of metritis treatment strategies on health, behavior, reproductive, and productive responses of Holstein cows.

Our objectives were to compare the effects of ceftiofur crystalline free acid (CCFA) and ampicillin trihydrate (AMP) treatments of cows diagnosed with metritis on uterine health, behavior, reproductive, and productive responses. A controlled randomized clinical trial was designed. Metritis was defined as vaginal discharge (VD) = 5 (fetid, watery, red/brown) within 21 d in milk (DIM) and rectal temperature (RT) <39.5°C, whereas VD = 5 and RT ≥39.5°C was defined as puerperal metritis. On the day of diagnosis (d 0), cows were paired by parity and severity of metritis (metritis vs. puerperal metritis) and assigned randomly to the AMP and CCFA treatments. Cows enrolled in the AMP (n = 308) treatment were moved to a nonsalable-milk pen, where they were treated once daily for 5 d, and were moved back to their original pen 72 h after the last treatment (d 7). Cows enrolled in the CCFA (n = 310) treatment remained in their original pen and received 2 treatments of CCFA, 72 h apart. Rectal temperature was measured daily from d 0 to 6 and on d 11. Vaginal discharge was evaluated on d 4, 6, and 11 to assess cure. Cure was defined as the absence of treatment with additional antimicrobial before experiment d 11, VD <5, and RT <39.5°C. Cows were examined at 28 ± 3 DIM for purulent VD (PVD) and at 35 ± 3 DIM for cytological endometritis. Pregnancy was diagnosed at 40 ± 3 and 60 ± 7 d after first and second artificial inseminations. Cure of metritis did not differ between treatments on d 11 (AMP = 64.6 ± 3.1, CCFA = 63.5 ± 3.1%). Cows treated with AMP had greater RT from experiment d 1 to 6 compared with cows treated with CCFA (AMP = 39.1 ± 0.02, CCFA = 39.0 ± 0.02°C). Cows in the AMP treatment had greater prevalence of PVD at 28 ± 3 DIM (AMP = 82.6 ± 2.3, CCFA = 74.4 ± 2.7%) and tended to have greater prevalence of cytological endometritis at 35 ± 3 DIM (AMP = 77.8 ± 6.2 vs. CCFA = 61.7 ± 7.5%) than CCFA-treated cows. Treatment did not affect the hazard of pregnancy among multiparous cows; however, among primiparous cows, CCFA treatment reduced the hazard of pregnancy and increased the median days to pregnancy (AMP = 145 vs. CCFA = 169 d). Finally, average daily milk yield up to 14 wk postpartum was not affected by treatment (AMP = 38.0 ± 0.4, CCFA = 37.5 ± 0.4 kg). We conclude from the current experiment that CCFA was more effective in reducing RT and improving uterine health of metritic cows; however, the improved hazard of pregnancy of primiparous cows treated with AMP is important and warrants further investigation.

Using chitosan microparticles to treat metritis in lactating dairy cows.

The main objective of this study was to evaluate the efficacy of intrauterine administration of chitosan microparticles (CM) in curing metritis in dairy cows. A secondary objective was to evaluate the effects of metritis treatments on milk yield, survival, and reproductive performance. Cows with a fetid, watery, red-brownish vaginal discharge were diagnosed with metritis. Holstein cows (n = 826) with metritis from 3 dairies located in northern Florida were blocked by parity (primiparous or multiparous) and, within each block, randomly assigned to one of 3 treatments: CM (n = 276) = intrauterine infusion of 24 g of CM dissolved in 40 mL of sterile distilled water at the time of metritis diagnosis (d 0), 2 (d 2), and 4 (d 4) d later; ceftiofur (CEF; n = 275) = subcutaneous injection of 6.6 mg/kg ceftiofur crystalline-free acid in the base of the ear at d 0 and d 3; Control (CON; n = 275) = no treatment applied at metritis diagnosis. All groups could receive escape therapy if condition worsened. Cure was considered when vaginal discharge became mucoid and not fetid. A group of nonmetritic (NMET; n = 2,436) cows was used for comparison. Data were analyzed by generalized linear mixed and Cox's proportional hazard models. Cows in CM and CON had lesser risk of metritis cure on d 12 than cows in CEF (58.6 ± 5.0 vs. 61.9 ± 4.9% vs. 77.9 ± 3.9, respectively). The proportion of cows culled within 60 days in milk (DIM) was greater for cows in CM than for cows in CEF and CON (21.5 ± 2.7 vs. 9.7 ± 1.9 vs. 11.3 ± 2.0%, respectively). Treatment did not affect rectal temperature or plasma nonesterified fatty acids, ß-hydroxybutyrate, and haptoglobin concentrations. Milk yield in the first 60 DIM differed for all treatments, and it was lowest for CM (35.8 ± 0.3 kg/d), followed by CON (36.8 ± 0.3 kg/d) and CEF (37.9 ± 0.3 kg/d). The hazard of pregnancy up to 300 DIM was lesser for CM than CEF (hazard ratio = 0.62; 95% CI: 0.50-0.76), for CM than CON (hazard ratio = 0.77; 95% CI: 0.62-0.95) and for CON than CEF (hazard ratio = 0.80; 95% CI: 0.65-0.99). Culling was greater, and milk yield and fertility were lesser for CEF than NMET. In summary, CM did not improve the cure of metritis, and was detrimental to milk yield, survival, and fertility compared with CON. In contrast, CEF increased the cure of metritis, milk yield, and fertility compared with CM and CON. Finally, the negative effects of metritis on milk yield culling and fertility could not be completely reversed by CEF.

Association of dry matter intake and energy balance prepartum and postpartum with health disorders postpartum: Part II. Ketosis and clinical mastitis.

The main objective of this study was to determine the association of dry matter intake as percentage of body weight (DMI%BW) and energy balance (EB) prepartum (-21 d relative to parturition) and postpartum (28 d) with ketosis (n = 189) and clinical mastitis (n = 79). For this, DMI%BW and EB were the independent variables and ketosis and clinical mastitis were the dependent variables. A secondary objective was to evaluate prepartum DMI%BW and EB as predictors of ketosis and clinical mastitis. For this, ketosis and clinical mastitis were the independent variables and DMI%BW and EB were the dependent variables. Data from 476 cows from 9 experiments were compiled. Clinical mastitis was diagnosed if milk from 1 or more quarters was abnormal in color, viscosity, or consistency, with or without accompanying heat, pain, redness, or swelling of the quarter or generalized illness, during the first 28 d postpartum. Ketosis was defined as the presence of acetoacetate in urine that resulted in any color change [5 mg/dL (trace) or higher] in the urine test strip (Ketostix, Bayer, Leverkusen, Germany). Cows that developed ketosis had lesser DMI%BW and lesser EB on d -5, -3, -2, and -1 than cows without ketosis. Each 0.1-percentage point decrease in the average DMI%BW and each 1-Mcal decrease in the average of EB in the last 3 d prepartum increased the odds of having ketosis by 8 and 5%, respectively. Cut-offs for DMI%BW and EB during the last 3 d prepartum to predict ketosis were established and were ≤1.5%/d and ≤1.1 Mcal/d, respectively. Cows that developed ketosis had lesser postpartum DMI%BW and EB and greater energy-corrected milk (ECM) than cows without ketosis. Cows that developed clinical mastitis had lesser DMI%BW but similar prepartum EB compared with cows without clinical mastitis. Each 0.1-percentage point decrease in the average DMI%BW and each 1-Mcal decrease in the average EB in the last 3 d prepartum increased the odds of having clinical mastitis by 10 and 8%, respectively. The average DMI%BW and EB during the last 3 d prepartum produced significant cut-offs to predict clinical mastitis postpartum, which were ≤1.2%/d and ≤1.0 Mcal/d, respectively. Cows that developed clinical mastitis had lesser postpartum DMI%BW from d 3 to 15 and on d 17; greater EB on d 18, from d 21 to 23, and on d 26; and lesser ECM. The main limitation in this study is that the time-order of disease relative to DMI%BW and ECM is inconsistent such that postpartum outcomes were measured before and after disease, which was diagnosed at variable intervals after calving. In summary, measures of prepartum DMI were associated with and were predictors of ketosis and clinical mastitis postpartum, although the effect sizes were small.

Association of dry matter intake and energy balance prepartum and postpartum with health disorders postpartum: Part I. Calving disorders and metritis.

The main objective of this study was to determine the association of dry matter intake as percentage of body weight (DMI%BW) and energy balance (EB) prepartum (-21 d relative to parturition) and postpartum (28 d) with calving disorders (CDZ; dystocia, twins, and stillbirths; n = 101) and metritis (n = 114). For this, DMI%BW and EB were the independent variables and CDZ and metritis were the dependent variables. A secondary objective was to evaluate prepartum DMI%BW and EB as predictors of CDZ and metritis. For this, CDZ and metritis were the independent variables and DMI%BW and EB were the dependent variables. Data from 476 cows from 9 experiments were compiled. Cows that developed CDZ had lesser postpartum DMI%BW from d 3 to 12 and lesser energy-corrected milk (ECM) than cows that did not develop CDZ. Dry matter intake as percentage of BW and EB prepartum did not affect the odds of CDZ. Cows with metritis had lesser prepartum DMI%BW and EB. Each 0.1-percentage point decrease in the average DMI%BW and each 1-Mcal decrease in the average EB in the last 3 d prepartum increased the odds of having metritis by 8%. The average DMI%BW and EB during the last 3 d prepartum produced significant cut-offs to predict metritis postpartum, which were ≤1.6%/d and ≤2.5 Mcal/d, respectively. Cows that developed metritis had lesser overall postpartum DMI%BW and ECM and lesser EB from d 2 to 5 and from d 7 to 11 than cows that did not develop metritis. The main limitation in this study is that the time-order of disease relative to DMI%BW and ECM is inconsistent such that postpartum outcomes were measured before and after disease, which was diagnosed at variable intervals after calving. In summary, prepartum DMI%BW and EB were associated with and were predictors of metritis although the effect sizes were small for metritis, and calving disorders and metritis were associated with decreased DMI%BW and ECM postpartum.

Rumination time and monitoring of health disorders during early lactation.

The objective was to evaluate the association between changes in daily rumination time (dRT) and early stages of disease during early lactation and to assess the performance of two proposed disease detection indices. This cohort study included 210 multiparous Holstein cows at the University of Florida Dairy Unit. Cows were affixed with a neck collar containing rumination loggers providing rumination time. The occurrence of health disorders (mastitis, metritis, clinical hypocalcemia, depression/dehydration/fever (DDF), digestive conditions, lameness and clinical ketosis) was assessed until 60 days in milk. Two indices were developed to explore the association between dRT and disease: (i) Cow index (CIx), based on changes in dRT in the affected cow during the days before the diagnosis of health disorders; (ii) Mates index (MIx), index based on deviations in dRT relative to previous days and healthy pen mate cohorts. Subsequently, an alert model was proposed for each index (ACIx and AMIx) considering the reference alert cut-off values as the differences between average index values in healthy and sick cows for each specific disease. The sensitivity (SE) of ACIx detecting disease ranged from 42% (digestive conditions and DDF) to 80% (clinical hypocalcemia) with 84% specificity (SP). The SE of AMIx ranged from 46% (digestive conditions and DDF) to 100% (clinical hypocalcemia) with 85% SP. Consistent reductions in rumination activity, both within cow and relative to healthy mate cohorts, were observed for each health disorder at the day of diagnosis. However, the ability of the proposed algorithms for detecting each specific disease was variable.

Scale-up of the electrokinetic fence technology for the removal of pesticides. Part II: Does size matter for removal of herbicides?

This work reports results of the application of electrokinetic fence technology in a 32 m3 -prototype which contains soil polluted with 2,4-D and oxyfluorfen, focusing on the evaluation of the mechanisms that describe the removal of these two herbicides and comparing results to those obtained in smaller plants: a pilot-scale mockup (175 L) and a lab-scale soil column (1 L). Results show that electric heating of soil (coupled with the increase in the volatility) is the key to explain the removal of pollutants in the largest scale facility while electrokinetic transport processes are the primary mechanisms that explain the removal of herbicides in the lab-scale plant. 2-D and 3-D maps of the temperature and pollutant concentrations are used in the discussion of results trying to give light about the mechanisms and about how the size of the setup can lead to different conclusions, despite the same processes are occurring in the soil.

Scale-up of the electrokinetic fence technology for the removal of pesticides. Part I: Some notes about the transport of inorganic species.

This work describes the application electrokinetic fence technology to a soil polluted with herbicides in a large prototype containing 32 m3 of soil. It compares performance in this large facility with results previously obtained in a pilot-scale mockup (175 L) and with results obtained in a lab-scale soil column (1 L), all of them operated under the same driving force: an electric field of 1.0 V cm-1. Within this wide context, this work focuses on the effect on inorganic species contained in soil and describes the main processes occurring in the prototype facility, as well as the differences observed respect to the lower scale plants. Thus, despite the same processes can be described in the three plants, important differences are observed in the evolution of the current intensity, moisture and conductivity. They can be related to the less important electroosmotic fluxes in the larger facilities and to the very different distances between electrodes, which lead to very different distribution of species and even to a very different evolution of the resulting current intensity. 2-D maps of the main species at different relevant moments of the test are discussed and important information is drawn from them. Ions depletion from soil appears as a very important problem which should be prevented if the effect of natural bioremediation and/or phytoremediation on the removal or organics aims to be accounted.

Clinical response after chitosan microparticle administration and preliminary assessment of efficacy in preventing metritis in lactating dairy cows.

The objectives were to evaluate the clinical response to intrauterine administration of chitosan microparticles (CM) and to assess efficacy for preventing metritis in dairy cows. Holstein cows (n=104; 40 primiparous and 64 multiparous) at increased risk for metritis (cows that had abortion, dystocia, twins, stillbirth, or retained placenta) were randomly assigned to 1 of 2 treatments at 1d in milk (DIM; 24h postpartum): CM group (n=52), daily intrauterine infusion of 8g of CM dissolved in 40mL of sterile water for 5d; control (CON) group (n=52), daily intrauterine infusion of 40mL of sterile water for 5d. Clinical response was assessed by evaluation of parameters associated with inflammation (rectal temperature and plasma haptoglobin concentration) and metabolism [plasma nonesterified fatty acid (NEFA) and ß-hydroxybutyrate (BHB) concentrations] up to 14 DIM, and daily milk yield up to 30 DIM. Uterine discharge pH was evaluated at 4, 7, 10, and 14 DIM as an indicator of bacterial load and acid byproduct production. The cumulative incidence of metritis was evaluated up to 4, 7, 10, and 14 DIM. Continuous and dichotomous outcomes were evaluated with mixed linear and logistic regression analysis, respectively. Treatment with CM did not affect rectal temperature (39.17±0.04 vs. 39.14±0.04°C), haptoglobin (1.10±0.05 vs. 1.07±0.05mg/mL), NEFA (0.64±0.04 vs. 0.63±0.04mmol/L), BHB (0.61±0.03 vs. 0.57±0.03mmol/L), or milk yield (30.3±0.92 vs. 30.1±0.97kg/d) compared with CON. An interaction between treatment and time showed that NEFA concentrations were lower for CM than CON at 10 DIM (0.46±0.06 vs. 0.64±0.06mmol/L). Treatment with CM resulted in greater uterine discharge pH than CON (6.91±0.03 vs. 6.83±0.02). Cows that developed metritis had increased concentrations of haptoglobin and BHB, and decreased uterine discharge pH and milk yield. Treatment with CM resulted in decreased incidence of metritis up to 7 DIM compared with CON (46.2 vs. 65.4%); however, no differences were found at 4 (11.5 vs. 17.3%), 10 (61.5 vs. 73.1%), and 14 DIM (63.5 vs. 73.1%) for CM versus CON, respectively. In conclusion, CM did not alter clinical parameters of cows at risk for metritis, and may merit further investigation for prevention of metritis. However, the duration of treatment may have to be extended to effectively reduce the incidence of metritis during the high-risk period.

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

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

Effects of oral calcium supplementation on productive and reproductive performance in Holstein cows.

The objectives of this experiment were to determine the effects of oral Ca supplementation on milk yield, body condition, pregnancy per artificial insemination (P/AI), and days to pregnancy in Holstein cows considered to be of low (LRM; no calving assistance, live singleton without retained placenta) or high (HRM; at least one of the following: dystocia, twins, stillbirth, retained placenta, or vulvo-vaginal laceration) risk of developing metritis. The hypotheses were that oral Ca supplementation during the early postpartum period would reduce the loss of body condition and improve lactation performance and reproduction. Four hundred fifty Holstein cows were blocked by parity as primiparous cows (n=174) or multiparous cows (n=276). Within parity, a cow considered at HRM was matched with a cow of LRM and the pair was randomly assigned to control (no Ca supplementation), 86g of Ca on d 0 and 1 postpartum (CaS1), or 86g of Ca on d 0 and 1 postpartum followed by 43g/d on d 2 to 4 postpartum (CaS4). Body condition was scored at calving and 32 d postpartum and estrous cyclicity was evaluated at 38 and 52 d postpartum. Milk yield was recorded daily. Multiparous cows were classified as above or below the mean 305-d mature equivalent milk yield based on production in the previous lactation. Reproductive performance was evaluated for the first 210 d postpartum. Body condition did not differ among treatments, and cows lost on average 0.44 units of body condition in the first month of lactation. Calcium supplementation did not affect milk yield in the first 5 mo postpartum. For multiparous cows, Ca supplementation was beneficial to milk yield in the first 30 DIM in cows of greater production potential, but detrimental to multiparous cows with below average production potential. Calcium supplementation to primiparous cows reduced P/AI at first (control=55.8, CaS1=31.5, CaS4=37.0%) and all artificial inseminations (control=48.5, CaS1=34.6, CaS4=38.5%); however, Ca supplementation to multiparous cows improved P/AI at the first (control=32.1, CaS1=38.6, CaS4=41.3%) and all artificial inseminations (control=28.1, CaS1=35.3, CaS4=40.5%). These responses in P/AI to Ca supplementation resulted in extended median days to pregnancy (control=75, CaS1=100, CaS4=94 d) and smaller proportion of pregnant cows (control=89.3, CaS1=83.9, CaS4=83.9%) in primiparous cows, but shorter days to pregnancy (control=115, CaS1=94, CaS4=94 d) and increased proportion of pregnant cows in multiparous cows (control=67.0, CaS1=77.2, CaS4=74.3%). Risk of metritis depressed most measures of reproduction evaluated in the experiment. Results indicate that responses to oral Ca supplementation are conditional on parity and production potential of cows. Oral Ca supplementation was detrimental to reproduction in primiparous cows. On the other hand, Ca supplementation benefited reproduction in multiparous cows and milk yield in the cohort of multiparous cows of greater production potential.

Effects of oral calcium supplementation on mineral and acid-base status, energy metabolites, and health of postpartum dairy cows.

Two experiments were conducted to characterize blood concentrations of minerals and acid-base status after oral dosing of Ca salts and to determine the effects of oral Ca on mineral and metabolic status and incidence diseases. The hypotheses were that administration of oral Ca as CaCl2 and CaSO4 maintains blood total Ca (tCa) concentrations ≥2.125 mM and reduces the incidence of diseases in early lactation. In experiment 1, 18 Holstein cows on the day of calving were assigned to receive a single dose of 0, 43, or 86g of Ca as an oral bolus. Blood was sampled before and after treatments to characterize acid-base status and concentrations of minerals. In experiment 2, 450 Holstein cows considered of low (LRM; normal calving) or high risk (HRM; dystocia, twins, stillbirth, retained placenta, vulvo-vaginal laceration, or a combination of these) of metritis (primiparous-LRM=84; primiparous-HRM=84; multiparous-LRM=138; multiparous-HRM=138) on the day of calving were blocked by parity and then randomly assigned to control, no Ca supplementation; 86g of Ca on d 0 and 1 postpartum (CaS1); or 86g of Ca on d 0 and 1 postpartum followed by 43g/d on d 2 to 4 postpartum (CaS4). Blood was sampled before and 30 min after treatment on d 0, and 30 min after treatments on d 1 to 4, and d 7 and 10 for determination of concentrations of minerals and metabolites and blood acid-base responses. Disease incidence was evaluated for the first 30 DIM. Concentrations of ionized Ca (iCa) increased for 2h in cows supplemented with 43g of Ca and fewer than 8h in cows supplemented with 86g of Ca. The changes in iCa concentrations from pretreatment to 30 min after 86g of Ca supplemented on d 0 were 0.11±0.03 mM in multiparous cows and 0.25±0.03 mM in primiparous cows. Oral Ca reduced the incidence of subclinical hypocalcemia (SCH; tCa <2.125mM) in the first 4 d in the experiment (control=69.3%; CaS1=57.5%; CaS4=34.2%). Calcium supplementation decreased the prevalence of SCH on d 0 and 1 postpartum in all cows. Stopping oral Ca in CaS1 on d 1 postpartum, however, caused a rebound in SCH on d 2 to 4 postpartum in primiparous cows. Oral Ca increased the incidence of metritis (control=22.7%; CaS1=34.8%; CaS4=32.8%), primarily because of an increase in LRM primiparous cows (control=17.9%; CaS1=35.7%; CaS4=42.9%). Oral Ca increased morbidity in primiparous cows (control=38.1%; CaS1=61.8%; CaS4=60.3%) but had no effect on multiparous cows (control=38.2%; CaS1=35.1%; CaS4=30.1%). Large doses of oral Ca as salts of chloride and sulfate in the first days postpartum should be avoided in primiparous cows and used only in cows at risk of clinical hypocalcemia.

Removal of oxyfluorfen from spiked soils using electrokinetic soil flushing with the surrounding arrangements of electrodes.

This work reports the results of a study in which the remediation of soil that undergoes an accidental discharge of oxyfluorfen is carried out by using electrokinetic soil flushing (EKSF). Two different electrode configurations were tested, consisting of several electrodes surrounding an electrode of different polarity (so-called 1A6C, one anode surrounded by six cathodes, and 1C6A, one cathode surrounded by six cathodes). A pilot plant scale was used (with a soil volume of 175dm(3)) to perform the studies. During the tests, different parameters were measured daily (flowrates, pH, electrical conductivity and herbicide concentration in different sampling positions). Furthermore, at the end of the test, a complete post-mortem analysis was carried out to obtain a 3-D map of the pollution, pH and electrical conductivity in the soil. The results demonstrate that electrode arrangement is a key factor for effective pollutant removal. In fact, the 1A6C configuration improves the removal rate by 41.3% versus the 27.0% obtained by the 1C6A configuration after a period of 35days. Finally, a bench mark comparison of this study of soil remediation polluted with 2,4-D allows for significant conclusions about the scale-up and full-scale application of this technology.

Vulvovaginal laceration as a risk factor for uterine disease in postpartum dairy cows.

The main objective was to evaluate the association between vulvovaginal laceration and uterine diseases in dairy cows. The secondary objectives were to evaluate the association between vulvovaginal laceration and cyclicity, and reproductive performance. The vulvovaginal region of 660 Holstein cows from a 5,000 lactating-cows herd was inspected at 4d in milk (DIM) for the presence of lacerations, and scored (VLS) as follows: 0=no laceration; 1=laceration <2cm at the dorsal commissure of the vulva or lateral walls of the vulva/vagina; 2=laceration ≥2cm at the dorsal commissure of the vulva or at the lateral walls of the vulva/vagina, or both. Vaginal discharge was scored at 4, 6, and 8 DIM for diagnosis of metritis, and then at 32 DIM for diagnosis of purulent vaginal discharge (PVD). Data were analyzed using LOGISTIC and PHREG procedures of SAS. Cows with VLS 2 had greater incidence of metritis than cows with VLS 0 (69.1 vs. 42.4%), and cows with VLS 1 tended to have greater incidence of metritis than cows with VLS 0 (52.0 vs. 42.4%). Cows with VLS 2 had greater incidence of PVD than cows with VLS 0 (56.5 vs. 43.1%). A lower proportion of cows with VLS 2 than VLS 0 were cyclic by 64 DIM (70.0 vs. 86.8%). A lower proportion of cows with VLS 2 than VLS 0 were pregnant at 60 d after first AI (28.7 vs. 33.6%). Proportion of pregnant cows at 60d after AI tended to be lower for VLS 1 than VLS 0 (28.4 vs. 33.6%). Hazard of pregnancy by 300 DIM was not affected by VLS. Hazard of pregnancy was decreased for cows with metritis, PVD, and anovular cows. In summary, vulvovaginal laceration was associated with uterine disease and cyclicity, which were negatively associated with reproductive performance. Vulvovaginal laceration was recognized as a risk factor for postpartum uterine disease.

Associations between pregnancy-associated glycoproteins and pregnancy outcomes, milk yield, parity, and clinical diseases in high-producing dairy cows.

Pregnancy-associated glycoproteins (PAG) are produced by the ruminant placenta and secreted into the maternal circulation throughout pregnancy. The extent to which circulating PAG concentrations predict pregnancy outcomes was examined herein. Also, associations between circulating PAG concentrations and various production parameters and clinical diseases were evaluated. Lactating primiparous and multiparous Holstein cows (n=345) were bred via timed artificial insemination using a standard Ovsynch protocol. Pregnancy was diagnosed by transrectal ultrasonography at d 32, 46, and 74 of gestation. Blood was harvested at d 32 to determine plasma concentrations of PAG and progesterone. Cows pregnant at d 32 that subsequently lost their pregnancy at d 46 and 74 had reduced PAG concentrations. Both artificial insemination service number and parity were associated with plasma PAG concentrations. Concentration of PAG in plasma was greater for cows pregnant from their second or later breeding than those pregnant from the first breeding postpartum, and was increased for primiparous compared with multiparous. In addition, cows with greater milk yield had increased plasma PAG concentrations. No association was detected between body condition score and plasma PAG concentrations. Cows that experienced clinical metritis, metabolic problems, or left displacement abomasum in the early postpartum period preceding breeding had greater plasma PAG concentrations than cows not experiencing these clinical diseases. Also, cows with multiple clinical diseases had increased odds of pregnancy loss when compared with cows not experiencing clinical diseases. Odds ratio testing detected a tendency in the relationship between reduced milk yield and increased pregnancy loss. Collectively, these associations illustrate one feature of the early developing placenta that may predict pregnancy outcomes in dairy cattle. It is unclear if plasma PAG are actively involved with mediating pregnancy outcomes, but modifications in circulating PAG concentrations due to pregnancy loss, milk yield, parity, and clinical disease implicate placental PAG production or PAG release as being responsive to various physiological stimuli.

Electrokinetic flushing with surrounding electrode arrangements for the remediation of soils that are polluted with 2,4-D: A case study in a pilot plant.

This work aimed to evaluate electrokinetic soil flushing (EKSF) technologies for the removal of 2,4-dichlorophenoxyacetic acid (2,4-D) from spiked soils using an electrode configuration consisting of one cathode surrounded by six anodes (1c6a) and one anode surrounded by 6 cathodes (1a6c). Experiments were conducted for over one month in a bench-scale set-up (175 dm(3) of capacity) that was completely automated and operated at a constant electric field (1.0V cm(-1)). The electrical current, temperature, pH, moisture and pollutant concentration in electrolyte wells were monitored daily, and at the end of the experiments, an in-depth sectioned analysis of the complete soil section (post-mortem analysis) was conducted. Despite the geometric similarity, the two strategies led to very different results mainly in terms of water and herbicide mobilization, whereas pH and conductivity do not depend strongly on the electrode configuration. The volume of water extracted from cathodes with 1a6c is seven times higher than that of the 1c6a strategy. Herbicide was transported to the anode wells by electromigration and then dragged toward the cathode wells by electro-osmotic fluxes, with the first process being much more important. The configuration 1c6a was the most efficient and attained a transfer of 70% of the herbicide contained in the soil to flushing water in 35 days. These results outperform those obtained by the configuration 1a6c, for which less than 8% of the herbicide was transferred to flushing fluids in a much longer time (58 days).