Associations of somatic cell count with milk yield and reproductive performance in grazing dairy cows.

Poor udder health status can have a detrimental effect on milk yield and reproductive performance, leading to reductions in the dairy farm profit. The objective of this retrospective longitudinal study was to assess the associations of somatic cell count (SCC) with daily milk yield and reproductive performance. A database with 1,930,376 lactations from 867 Argentinean grazing dairy herds records collected for 14 years was used. The association of the evolution of SCC (healthy vs. new case vs. cured vs. chronic; with 150,000 SCC/mL as threshold) and of the severity of SCC [mild (150,000--400,000 SCC/mL) vs. moderate (400,000-1,000,000 SCC/mL) vs. severe (>1,000,000 SCC/mL)] with the odds for conception were estimated. Finally, the associations of the linear score of SCC (LS-SCC) with daily milk yield were estimated depending on parity and milk production quartile. The odds ratios (CI 95%) for conception at first service were 0.921 (0.902-0.941), 0.866 (0.848-0.884), and 0.842 (0.826-0.859) for the new case, cured, and chronic cows compared with healthy cows, respectively. Also, the odds ratios (CI 95%) for conception were 0.902 (0.881-0.925), 0.837 (0.808-0.866) and 0.709 (0.683-0.736) for mild, moderate and severe cases compared with healthy cows, respectively. An increase of one point of LS-SCC was associated with decreases of 0.349, 0.539, and 0.676 kg in daily milk yield for first-, second-, and third-lactation cows, respectively. In conclusion, SCC is negatively associated with the risk for conception and with daily milk yield in grazing dairy cows. This negative relationship with conception is higher when SCC increase occurs after the service date and it is influenced by severity of mastitis, and in the case of milk yield, the negative association is influenced by parity, milk production quartile, and severity of mastitis.

Association between progesterone concentration and endometrial gene expression in dairy cows.

The objective of this study was to evaluate the association between progesterone concentration on Days 4 and 9 of the estrus cycle and endometrial transcriptome at Day 9 in lactating grazing dairy cows. Blood samples were obtained on Days 0, 4, and 9 for progesterone measurement by chemiluminescence. Cows were assigned to one of the following groups (n = 3 per group): cows with low physiological progesterone on Day 4, cows in anestrous, cows with high physiological progesterone on Day 4, and superovulated cows. Endometrial biopsy samples were obtained on Day 9 for RNA sequencing. Quality control and determination of differentially expressed genes (false discovery rate <0.05) were determined using the edgeR package for R software. We identified 3,042 differentially expressed genes among the 4 groups. Cows having high physiological progesterone and superovulated cows showed high similarities and clustered apart from those in anestrus or having low physiological progesterone. Functional analysis using Database for Annotation, Visualization, and Integrated Discovery revealed that endometrial genes upregulated by low progesterone concentration are enriched genes involved in the immune system and inflammatory response. Conversely, cows with high physiological progesterone concentration presented an endometrial transcriptome with similarities to cows with good genetic merit for fertility, showing upregulation of genes related to uterine relaxation-contraction, focal adhesion, GnRH signaling pathway, and epidermal growth factor-like related terms, suggesting a favorable embryo environment. In conclusion, our results support the concept that there is a threshold of progesterone concentration at the beginning of the luteal phase associated with endometrial expression of critical genes involved in the preparation of the uterine environment for embryo implantation.

Association between prepartum metabolic status and resumption of postpartum ovulation in dairy cows.

Cows transitioning from late gestation to early lactation experience an increase in energy demands, which lead to a negative energy balance (NEB) because the greater energy requirement is not fully synchronized with the intake of dry matter. In this context, there is an increase in plasma NEFA and ghrelin concentrations and a decrease in plasma insulin and glucose-dependent insulinotropic polypeptide (GIP) concentrations. This situation could have a negative impact on the return to cyclicity because some of these variables have been associated with reduced GnRH and LH pulsatility (high NEFA and low insulin concentrations). However, there are no studies showing the relationship between ghrelin or GIP and reproductive performance. It is known that these hormones are related with lipolysis and NEB, with NEB being one of the main determinants of GnRH pulse generator activity. Thus, the objective of the present study was to evaluate the association between plasma NEFA concentration and metabolic hormones (insulin, ghrelin, and GIP) before parturition and their associations with the resumption of postpartum ovulations in dairy cows. A completely randomized block design was used in a commercial dairy herd with sampling day (visit to farm) as the blocking criteria. Holstein cows (n = 92) were screened for plasma NEFA concentration -5 d (±2 d) relative to the expected parturition day, and top and bottom quartiles were considered as high (H-NEFA) and low (L-NEFA) NEFA groups. Data were analyzed with correlation, linear regression, and proportional hazard regression models. Plasma NEFA concentration (H-NEFA mean = 294 µM, SD = 141.2; and L-NEFA mean = 122 µM, SD = 25.3) was correlated (P < 0.01) with plasma insulin (r = -0.374) and ghrelin (r = -0.346) concentrations but not with plasma GIP concentration (P = 0.64). The greater the concentration of insulin, the lesser the prepartum NEFA concentration (for each 1 µU/mL of plasma insulin increase, there is a decrease of 1.223 ± 0.62 µM of NEFA). Plasma ghrelin and GIP concentrations were not associated with plasma NEFA concentration. Finally, H-NEFA prepartum cows were less likely to resume ovulation than L-NEFA cows (hazard ratio [HR] = 0.563, 95% confidence interval [CI] = 0.314-1.011), whereas high ghrelin cows were more likely to resume ovulation than low ghrelin cows (HR = 1.873, 95% CI = 0.846-4.145). Conversely, resumption of ovulation was not associated with prepartum insulin and GIP concentrations. Prepartum NEFA and possibly ghrelin are associated with the return to postpartum cyclicity; however, insulin and GIP are not related to the resumption of ovulation in dairy cows.

Parenteral Zinc Supplementation Increases Pregnancy Rates in Beef Cows.

Zinc (Zn) is required for normal reproductive performance in cattle. The aim of this study was to evaluate the effect of subcutaneous injection of 400 mg Zn at the beginning of fixed-time artificial insemination (FTAI) on preovulatory follicle and corpus luteum (CL) size, plasma estradiol (E2) and progesterone (P4) concentrations, and pregnancy rates in beef cows. Copper (Cu) concentration and alkaline phosphatase (ALP) activity in plasma were also evaluated. Zinc supplementation at the beginning of the FTAI protocol (day 0) increased the area of preovulatory follicle (APF, day 9; P = 0.042) and plasma P4 concentration (day 16; P = 0.01), whereas plasma E2 concentration (day 9) and area of CL (ACL; day 16) were not modified by Zn supplementation in cows with adequate plasma Zn concentration. Zinc supplementation in Zn-deficient cows increased ACL with respect to controls (P = 0.048) but did not modify plasma E2 concentration. Pregnancy rate on day 41 after FTAI was higher in cows supplemented with Zn compared with controls (80.95% and 51.61%, respectively; P = 0.042). Plasma Zn and Cu concentrations on days 7, 9, and 16 were not affected by Zn supplementation. In conclusion, the results obtained in the present study determined that parenteral Zn supplementation at the beginning of the FTAI protocol increased preovulatory follicle size, plasma P4 concentration, and pregnancy rates in beef cows.

Effect of milk production on reproductive performance in dairy herds.

The objective of the present study was to assess the relationship between individual cow milk yield and fertility, accounting for the contextual effect of the herd. A data set including 657,968 lactations from 677 dairy herds in Argentina from 2001 to 2012 was used. The odds of pregnancy by 100 d in milk (DIM) were assessed by a multilevel logistic model (with cow as the first and herd as the second hierarchical level), and time to pregnancy was assessed by a proportional hazards regression model. Multilevel logistic models included the fixed effects of milk yield by 80 DIM, parity, year, and calving season at cow level and quartiles of herd milk yield by 80 DIM as a contextual effect. The proportional hazards model included the effect of daily cow-level milk yield as time-dependent variable, with milk yield at herd level as the stratification variable. Cows producing 1 standard deviation over the mean milk yield of their herd had 1.3 percentage point lower pregnancy by 100 DIM (from 31.4 to 30.1%; odds ratio = 0.942) when in herds in the top quartile of milk yield, whereas they increased 0.5 percentage points (from 27.9 to 28.4%) when in herds in the lowest quartile of milk yield. Only 4% of the observed variation in pregnancy by 100 DIM was explained by the random effect of the herd. Similarly, cows producing 1 standard deviation (8 kg/d) greater than the herd mean daily milk had 1.3% lower hazard of pregnancy (hazard ratio = 0.987) at 63 DIM in herds in the top quartile of milk yield, whereas they had 14.8% higher hazard (hazard ratio = 1.148) in herds in the lowest quartile of milk yield. The magnitude of the negative association between the cow's daily milk yield and the hazard of pregnancy increased with DIM. In conclusion, the relationship between milk yield and reproductive performance is statistically significant, but the effect size is practically small and is modulated by herd production level.

Purulent vaginal discharge in grazing dairy cows: Risk factors, reproductive performance, and prostaglandin F2α treatment.

The objectives of this study were to assess the association of a 4-point scale of vaginal discharge score (VDS) with time to pregnancy to define criteria for a practical case of purulent vaginal discharge (PVD) in dairy cows, to test the risk factors for PVD, and, finally, the effect of a dose of PGF2α on cure and reproductive performance. In experiment 1, grazing Holstein cows (n = 2,414) had their vaginal discharge scored at ∼32 d in milk (DIM) on a 4-point scale, the effect of VDS on the hazard of pregnancy by 300 DIM was then assessed to derive a case definition of PVD. Risk factors for PVD and self-cure were also assessed. In experiment 2, grazing Holstein cows (n = 6,326) from 5 herds were checked for PVD at ∼30 DIM. Cows with PVD were assigned to receive one dose of 500 µg of PGF2α analog (Cloprostenol; Ciclase, Syntex SA, Buenos Aires, Argentina) per cow (odd ear tag number) or to remain untreated (even tag number). Cure was declared if cows presented clear normal vaginal discharge (VDS-0) at visit 2 (∼62 DIM). Data were analyzed with Cox's regression and mixed logistic models. In experiment 1, cows with VDS ≥1 had lower hazard of pregnancy and longer calving to pregnancy interval than cows with VDS-0. This finding was not affected by the time at which the diagnosis was performed. Therefore, a cow ≥21 DIM and having VDS ≥1 was used to define a case of PVD. The odds of PVD were greater in primiparous cows compared with multiparous, in cows with abnormal calving compared with those with normal calving, and in those losing BCS peripartum. In experiment 2, PGF2α treatment tended to slightly increase the hazard of pregnancy (adjusted hazard ratio = 1.13). Conversely, PGF2α had no effect on the odds of cure of PVD [adjusted odds ratio (AOR) = 1.19], pregnancy at first service (AOR = 1.03), or pregnancy by 100 DIM (AOR = 0.89) or 200 DIM (AOR = 1.27). In conclusion, cows with VDS ≥1 can be considered to have PVD because of their lower hazard of pregnancy and longer calving to pregnancy interval (up to 48 d). Important risk factors are parity, calving, and body condition score loss peripartum. Optimal time of diagnosis is ≥28 to 35 DIM because cows experience a high self-cure rate. Self-cure is also affected by parity, prepartum BCS, and VDS. Finally, as treatment with one dose of PGF2α had a small effect on the hazard of pregnancy and no effect on clinical cure, its therapeutic use in grazing dairy cows with PVD is not recommended.

Short communication: Repeat breeder cows with fluid in the uterine lumen had poorer fertility.

The objective of the present study was to test the association between the presence of fluid in uterine lumen and the fertility of repeat breeder cows (RBC). Grazing Holstein cows that were clinically normal and were not pregnant after at least 3 consecutive inseminations were defined as RBC and included in the study (n = 358). A cut-point ≥2 mm of uterine lumen visible by ultrasonography was used as indicative of the presence of fluid in the uterus. The effect of this fluid on the fertility of RBC was assessed by mixed logistic regression. Twenty percent of the RBC had ≥2 mm of fluid in their uterus, and this finding was associated with a marked reduction in the odds of conception (odds ratio = 0.46). Additional intrauterine sampling revealed that 24.0% of RBC showed signs of subclinical endometritis [≥5% neutrophils in endometrial cytology (SCE)]. However, we found no agreement between the finding of uterine fluid and the diagnosis of SCE (kappa coefficient = 0.03, 95% confidence interval = -0.07 to 0.14). In conclusion, the presence of fluid (≥2 mm) in uterine lumen, detected by ultrasonography, was associated with lower fertility in RBC, but was not related to the diagnosis of SCE. Thus, ultrasonographic evaluation is an important tool to assist in assessment of RBC.

Application of a bacteriological on-farm test to reduce antimicrobial usage in dairy cows with purulent vaginal discharge.

The objective of this study was to assess the effect of a selective antibiotic treatment strategy based on a quick bacteriological on-farm test (Petrifilm, 3M Corp., St. Paul, MN) compared with the conventional antibiotic treatment of all cows having clinical endometritis (CE) defined by the presence of purulent vaginal discharge on both clinical cure rate and reproductive performance. The study was simultaneously conducted with dairy cows reared under a highly supplemented rotational grazing system in Argentina and in a freestall system in Slovakia. Cows having an abnormal vaginal discharge (VD, indicative of clinical endometritis) on 21 to 35 d in milk (DIM) were randomly allocated to 1 of 2 study groups: selective treatment (ST) or conventional treatment (CT). All cows in the CT group (n = 174) received a single intrauterine administration of 500 mg of cephapirin. In the ST group (n = 178), treatment decision was made according to the results of the bacteriological on-farm test. For this test, we collected intrauterine samples with the cytobrush technique and stroke the brushes onto 2 different Petrifilm plates, one for aerobic count and another for Enterobacteriaceae count, incubated the plates, and counted the number of colonies after 24 h. Positive cows (≥5 colonies in one or both plates) received a single intrauterine treatment with 500 mg of cephapirin, whereas negative cows (<5 colonies) remained untreated. Clinical cure rate was assessed by direct vaginal inspection at 14 d after treatment (VD-0). The odds for conception at first artificial insemination, artificial insemination by 80 DIM, pregnancy by 100 DIM, and for nonpregnancy by 200 DIM were estimated with mixed logistic regression models. The hazard of conception was also assessed with proportional hazard regression model. The selective antibiotic treatment strategy based on the outcome of Petrifilm test reduced the number of required treatments (57%) and maintained similar efficacy in terms of clinical cure and reproductive performance as the conventional antibiotic treatment of all endometritic cows.

Uterine infection with bovine herpesvirus type 4 in dairy cows.

Diseases of the reproductive tract are a frequent problem in dairy herds. Herpesviruses are uterine pathogens also involved in other clinical diseases; for example, bovine herpesvirus type 4 BoHV-4 induces abortion, enteritis, metritis, pneumonia and vaginitis, but it can also be detected in healthy cows. The role of BoHV-4 in the development of clinical endometritis (CE) or subclinical endometritis (SE) has not clearly been described. Therefore, the objective of this study was to describe the prevalence of uterine BoHV-4 infection and its relationship with clinical, bacteriological and cytological findings in dairy cows 20-30 days after calving. The experiment was performed as a completely randomized block design, with farm (n = 10) as blocking criterion and with cow (n = 397) as the experimental unit. Logistic regression models were used to assess the effect of BoHV-4 infection on CE, SE and reproductive performance. Proportion of cows infected with BoHV-4 was 5.8% (n = 23/397). BoHV-4 was isolated in 11.0% (n = 12/109), 4.8% (n = 4/84) and 3.6% (n = 7/194) of cows diagnosed as CE, SE or healthy, respectively. A logistic model revealed that BoHV-4 infection showed a tendency to increase the risk for CE (AOR = 2.17; p = .10) but significantly reduced both, the odds for artificial insemination within 80 days post-partum (dpp) (AOR = 0.37; p = .035) and for pregnancy within 200 dpp (AOR = 0.13; p = .004). Furthermore, BoHV-4 infection increased the chance for intrauterine infection with Trueperella pyogenes (AOR = 5.55; p < .001) and vice versa (AOR = 5.79, p < .001). In conclusion, BoHV-4 infection is associated with reduced chances for insemination and pregnancy by 200 dpp and showed a trend to be associated with increased risk for CE. Furthermore, BoHV-4 and Trueperella pyogenes infections are strongly related.

Endometrial cytology, biopsy, and bacteriology for the diagnosis of subclinical endometritis in grazing dairy cows.

The objectives of this study were to assess the agreement between endometrial cytology and uterine biopsy for the diagnosis of subclinical endometritis (SEND) in grazing dairy cows, the interobserver agreement of the biopsy's readings, and the bacterial population isolated from the uterus of cows having SEND. In experiment 1, lactating Holstein cows (n=44) 31 to 59 d in milk (DIM) at sampling were enrolled. Clinical endometritis was diagnosed by direct evaluation of vaginal discharge and SEND by endometrial cytology evaluation. Two hundred cells per smear were counted to determine the percentage of polymorphonuclear neutrophilic leukocytes (PMNL). Cut-off values used were ≥8% PMNL at ≤33 DIM, ≥6% PMNL at 34 to 47 DIM, and ≥4% PMNL at ≥48 DIM. Biopsies were assessed blindly by 2 observers who categorized them into 4 groups according to their inflammatory changes: none, minimal, moderate, and severe inflammatory changes. Data were analyzed using the kappa coefficient and logistic regression. In experiment 2, lactating Holstein cows (n=60) 21 to 62 DIM were enrolled. Clinical endometritis and SEND were diagnosed as previously described. Samples were cultured for aerobic and anaerobic bacteria by routine methods of bacteriological testing. Data were analyzed with logistic regression. In experiment 1, little agreement was observed between cytology and biopsy outputs (kappa=0.151), and strong agreement between the 2 operators (kappa=0.854). The likelihood of having a normal biopsy (no inflammatory change) was greater for healthy cows than for those having SEND (odds ratio=13.145). The probability for getting normal uterine tissue decreased 2.1% for every increasing percentage point in PMNL. In experiment 2, no bacteria were isolated from cows with SEND, coagulase-negative staphylococci were commonly isolated from healthy cows, and Trueperella pyogenes was frequently isolated from cows with clinical endometritis. The likelihood of isolating T. pyogenes from uterine samples increased with the percentage of PMNL (odds ratio=1.100). In conclusion, biopsy showed low agreement with cytology for the diagnosis of SEND. Nevertheless, fertility trials using uterine biopsies to predict pregnancy outcomes are needed to determine its diagnostic usefulness. Finally, bacteriology would not be recommended as a diagnostic tool because no bacteria were isolated from cows with SEND.

The relationship between endometrial cytology during estrous cycle and cutoff points for the diagnosis of subclinical endometritis in grazing dairy cows.

The objectives of this study were to assess the effect of the stage of estrous cycle on the percentage of endometrial polymorphonuclear cells (PMN) obtained by cytobrush to determine cutoff values for the diagnosis of subclinical endometritis under pastoral conditions, to measure the prevalence of subclinical endometritis 21 to 62d in milk (DIM), and to evaluate the effect of subclinical endometritis on reproductive performance in grazing dairy cows. The first experiment was conducted on a commercial dairy farm in Buenos Aires province (Argentina), where 17 postpartum cyclic dairy cows without clinical endometritis were selected and synchronized by Ovsynch protocol. Endometrial cytology (cytobrush technique) and blood (tail vessels) samples were obtained on d0, 4, 11, and 18 of the estrous cycle (corresponding to estrus, metestrus, diestrus, and proestrus, respectively) and used for measuring percentage of PMN and P4 concentration, respectively. The percentage of PMN was determined 3times by blinded count by 2 operators. Data were analyzed with PROC MIXED, PROC GENMOD, and PROC FREQ from SAS 9.1. The percentage of PMN did not vary with the stage of the estrous cycle. In addition, PMN counts were below any of the reported thresholds in this study (4%) for most of the cows. Therefore, the risk for false positive test results as a consequence of physiological changes in the counts of PMN during estrous cycle is low. The second experiment was conducted on 4 commercial dairy farms in Buenos Aires province (Argentina), where lactating Holstein dairy cows (n=418) 21 to 62 DIM without clinical endometritis were studied. Samples of endometrial cytology were collected with the cytobrush technique. Data were analyzed with receiver operator characteristic curves with Sigmaplot 10.0, and with PROC GLIMMIX, PROC PHREG, and PROC LIFETEST from SAS 9.1. Cutoff values for the diagnosis of subclinical endometritis in grazing dairy cows are 8% PMN for 21 to 33 DIM, 6% PMN for 34 to 47 DIM, 4% PMN for 48 to 62 DIM, and overall 5% PMN for 21 to 62 DIM; the prevalence of subclinical endometritis 21 to 62 DIM was 17%. Finally, subclinical endometritis diagnosed at 21 to 62 DIM decreases the hazard for pregnancy (hazard ratio=0.668; 95% confidence interval=0.492-0.909) and increases the calving to conception interval by d30 compared with normal cows (median 95% confidence interval=133 vs. 93, respectively).

Metritis in dairy cows: risk factors and reproductive performance.

The objectives of this study were to assess the risk factors for metritis, its effects on milk yield and on reproductive performance, and the efficacy of ceftiofur therapy in Holstein dairy cows. Cows (n=303) from a commercial dairy herd in Argentina were studied. Cows were scored for body condition, and blood samples were collected on d -14, 7, 21, 31, 41, and 50 relative to parturition. Cows having a watery, purulent, or brown, and fetid vaginal discharge (VD) and rectal temperature ≤ 39.2°C were diagnosed as having clinical metritis, and those having a similar VD and rectal temperature >39.2°C were diagnosed as having puerperal metritis. Both clinical and puerperal metritis cows were randomly assigned to control (no treatment) or ceftiofur group (2.2mg/kg×3 consecutive days). Cure was declared if clear VD was observed at 21 d in milk (DIM). Blood samples were analyzed for nonesterified fatty acids, ß-hydroxybutyrate, and blood urea nitrogen using commercial kits, and for insulin-like growth factor-1, insulin, and leptin by RIA. Data were analyzed with PROC MIXED, GENMOD, PHREG, and LIFETEST from SAS (SAS Institute Inc., Cary, NC). The risk for metritis increased with dystocia, retained fetal membranes, and dead calf [AOR (adjusted odds ratio)=2.58, 95% CI: 1.189-5.559], and as prepartum nonesterified fatty acids levels increased (AOR=1.001, 95% CI: 0.999-1.002). Conversely, risk decreased as prepartum insulin-like growth factor-1 increased (AOR=0.65, 95% CI: 0.349-1.219). Cows having either clinical or puerperal metritis produced less milk by 90 DIM than did healthy cows (2,236 ± 172 vs. 2,367 ± 77 vs. 2,647 ± 82 kg, respectively). Cows with puerperal metritis had lower risk for pregnancy by 100 DIM (AOR=0.189, 95% CI: 0.070-0.479) and a lower hazard rate for pregnancy by 150 DIM (hazard rate: 0.753, 95% CI: 0.621-0.911), and took longer to get pregnant (129 vs. 111 vs. 109 d, for puerperal metritis, clinical metritis, and healthy cows, respectively). Ceftiofur treatment was not associated with cure rate or milk yield but was related to increased risk for pregnancy at timed artificial insemination (AOR=2.688, 95% CI: 0.687-10.832), and for lower risk of reproductive cull (AOR=0.121, 95% CI: 0.014-1.066). In conclusion, abnormal calving and negative energy balance are associated with increased risk for metritis. Metritis, especially puerperal metritis, correlates with reduced milk production and poor reproductive performance. Finally, the likelihood for having a normal VD (indicative of cure) increased 2.6% for every day of increase in postpartum time and was 2 times higher for cows with clinical metritis than for those with puerperal metritis.

Clinical endometritis in an Argentinean herd of dairy cows: risk factors and reproductive efficiency.

The objectives of this study were to assess the clinical and metabolic risk factors for clinical endometritis, the likelihood for having a normal vaginal discharge during postpartum, and the effects of endometritis on milk yield, reproductive efficiency, and metabolic status in Holstein cows. The study was conducted in a commercial dairy herd (Cordoba, Argentina) where 303 Holstein cows were enrolled. Cows were body condition scored (1 to 5) and tail bled on -14, 7, 21, 31, 41, and 50 d relative to parturition. Cows having a vaginal discharge with presence of pus between 21 and 41 d postpartum (dpp) were diagnosed as having clinical endometritis. Plasma blood samples were analyzed for nonesterified fatty acids (NEFA), ß-hydroxybutyrate (BHBA), and blood urea nitrogen using commercial kits and insulin-like growth factor 1, insulin, and leptin by RIA. Data were analyzed with PROC MIXED, PROC GENMOD, and PROC PHREG of SAS (SAS Institute Inc., Cary, NC). Abnormal calving and puerperal metritis increased the risk for endometritis [adjusted odds ratio (AOR)=2.21 for both]. High prepartum NEFA and high postpartum BHBA increased the risk for endometritis (AOR=1.003 and 1.001, respectively), whereas high prepartum blood urea nitrogen reduced it (AOR=0.853). Cut-offs of 456.6 µM NEFA and 402.5 µM BHBA had sensitivities of 0.69 and 0.58, and specificities of 0.88 and 0.86, respectively. The likelihood for having normal vaginal discharge increased with time (∼1% × dpp) and with normal calving. Cows with endometritis had higher milk yield than normal herdmates (27.8±0.9 vs. 25.7±0.4 kg/d), lower risk for pregnancy by 100 dpp (AOR=0.10), higher nonpregnancy risk by 200 dpp (AOR=2.87), and higher risk for culling than normal cows (AOR=2.28). Cows with endometritis had a lower hazard rate (0.44) for pregnancy and had approximately 70 d longer calving-to-conception intervals. Finally, endometritis had no effect on metabolic hormones. In conclusion, the risk for clinical endometritis increases with abnormal calving and puerperal metritis, as prepartum NEFA and postpartum BHBA concentrations increase. Prepartum NEFA and postpartum BHBA could be useful for the prediction of endometritis. Last, clinical endometritis has detrimental effects on reproductive efficiency, as affected cows take longer to get pregnant and are at higher risk for culling.

Acute copper intoxication after a Cu-Ca EDTA injection in rats.

It has been claimed that Cu-Ca EDTA releases the Cu slowly into the blood when injected as a therapeutic agent for Cu deficiency, but when overdosing occurs, animals may die in a few hours after the injection, suggesting that Cu release is faster than expected. A total of 30 rats were divided into six equal groups and injected with Cu-Ca EDTA at 4, 6, 13, 33 or 64 mg/kg body weight, the sixth group acted as control. All rats were sacrificed 24 h later, with the exception of rats from groups 33 and 64 that died 7 and 2 h postinjection respectively. The rate of Cu transference to the liver increased linearly with the dose, from (X +/- S.E.) 14.88 +/- 3.14 in group 4, to 134.47 +/- 21.86 microg/h in group 64. Injected animals developed liver and kidney degeneration, hemorrhage and necrosis, which increased in severity with the Cu dose. Liver Cu accumulation was highest for group 13 (866.8 +/- 84.23 ppm DM), while in group 64 reached only 167.0 +/- 14.99 ppm. It is concluded that during acute Cu poisoning, the rate of transfer is responsible for the liver and kidney damage, rather than the total amount of transferred Cu to those organs, as it is the case in chronic Cu poisoning. The high rate of Cu transfer possibly surpasses the intrinsic mechanisms of Cu deposition and free Cu is left into the cell with the development of necrosis.