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.     

Cytological endometritis at artificial insemination in dairy cows: Prevalence and effect on pregnancy outcome

The aims of our field study in dairy cows were (1) to consolidate cytotape (CT) as a valid technique to diagnose cytological endometritis (CYTO) during artificial insemination (AI); (2) to establish a cutoff point concerning the polymorphonuclear cells (PMN) proportion to diagnose CYTO at AI; (3) to assess the prevalence of CYTO at AI; and (4) to evaluate the effect of CYTO on the pregnancy outcome of that AI. The investigation was performed using 1,625 AI-CT samples harvested from 873 Holstein-Friesian cows from 18 dairy farms in the Flemish region of Belgium. The CT device consisted of adapting a 1.5-cm piece of paper tape on the top of a conventional AI catheter covered with a double guard sheet, allowing an endometrial cytology sample to be taken when performing an AI. A receiving operator characteristic curve was built to assess the threshold level above which the PMN proportion significantly affected the AI success. Multilevel generalized mixed-effect models were built to identify factors affecting the pregnancy outcome of the AI under investigation. Only 7 samples (0.4%) harvested in 5 cows were discarded because of low-quality parameters. The cutoff point for CYTO at AI was set at ≥1% PMN (sensitivity = 33.8%, specificity = 88.6%). Prevalence of CYTO at AI was 27.8%. The conception rate for CYTO-positive samples was 32.7%, whereas it was 47% for CYTO-negative samples. A CYTO-negative AI had 1.8 [odds ratio (OR)] more chances to become pregnant than a CYTO-positive one. Other factors identified as detrimental for the pregnancy outcome were body condition score ≤1.5 (OR = 0.6), relative 305-d milk yield (OR = 0.9), dystocia (OR = 0.3), parity ≥2 (OR = 0.7), and warm months of the year. In conclusion, CT is a consolidated technique to diagnose CYTO at AI, PMN 1% is the threshold level to diagnose CYTO at AI, around one-quarter of inseminated uteri suffer from CYTO, and affected uteri having a significantly lower chance to become pregnant from that insemination.     

Assessment of the main pathogens associated with clinical and subclinical endometritis in cows by culture and MALDI-TOF mass spectrometry identification

Clinical endometritis (CE) and subclinical endometritis (SCE) are diseases that affect dairy cows during the puerperium, causing negative effects on the animals' milk production and fertility. The objective of this study was to assess the main bacteria related to cases of CE and SCE from uterine samples of dairy cows in Brazilian herds. Selective and differential media were used for isolation of aerobic and anaerobic bacteria and further MALDI-TOF mass spectrometry (MS) identification. A total of 279 lactating dairy cows with 28 to 33 d in milk from 6 commercial farms were evaluated. Initially, cows were classified in 3 groups: cytologic healthy cows (n = 161), cows with CE (n = 83), and cows with SCE (n = 35). Healthy animals presented 97 species, followed by the CE group with 53 identified species, and SCE cows presented only 21 bacterial species. We found a significantly higher isolation rate of Trueperella pyogenes in CE (26.5%) cows compared with healthy and SCE cows. Some anaerobic species were exclusively isolated from the CE group, even though they presented lower frequency. Interestingly, 18.1% of samples from CE cows and 40% of SCE cows were negative to bacterial isolation. Despite the use of culture-dependent methods instead of molecular methods, the present study enabled the identification of a complex community of 127 different species from 48 genera, composed of aerobic and anaerobic bacterial species among the 3 different animal groups. The method of sample collection, culture, and identification by MALDI-TOF MS were essential for the success of the analyses.     

Subclinical hypocalcemia,plasma biochemical parameters,lipid metabolism,postpartum disease,and fertility in postparturient dairy cows

A study was conducted to evaluate the potential association between Ca status at calving and postpartum energy balance, liver lipid infiltration, disease occurrence, milk yield and quality parameters, and fertility in Holstein cows. One hundred cows were assigned to 1 of 2 groups based on whole-blood ionized Ca concentration ([iCa]) on the day of calving [d 0; hypocalcemic [iCa] <1.0 mmol/L (n = 51); normocalcemic [iCa] ≥1.0 mmol/L (n = 49)]. Cows were blocked based on calving date and parity. Blood samples were collected approximately 14 d from expected calving date (d −14), the day of calving (d 0), and on d 3, 7, 14, 21, and 35 postpartum for measurement of plasma nonesterified fatty acid, iCa, total Ca, glucose, and total and direct bilirubin concentrations, and plasma aspartate aminotransferase and gamma glutamyl transferase activities. Liver biopsies were obtained from a subset of cows on d 0, 7, and 35 for quantification of lipid content. Milk samples were collected on d 3, 7, 14, 21, and 35 postpartum for measurement of somatic cell count and percentages of protein, fat, and solids-not-fat. Data for peak test-day milk yield, services per conception, and days open were obtained from Dairy Herd Improvement Association herd records. Disease occurrence was determined based on herd treatment records. Hypocalcemic cows had significantly higher nonesterified fatty acids on d 0. Hypocalcemic cows also had significantly more lipid in hepatocytes on d 7 and 35 postpartum. However, no statistically significant differences were observed between groups for plasma aspartate aminotransferase and gamma glutamyl transferase activities or total and direct bilirubin concentrations. Milk protein percentage was lower in hypocalcemic cows on d 21 and 35. However other milk quality variables (somatic cell count, milk fat percentage, and solids-not-fat) and milk yield variables (peak test-day milk yield and 305-d mature-equivalent 4% fat-corrected milk yield) did not differ between groups. No differences were observed between groups in the occurrence of clinical mastitis, ketosis, displaced abomasum, dystocia, retained placenta, metritis, or fertility measures (percentage cycling at 50–60 d postpartum, services per conception, or days open). These data suggest that early lactation fatty acid metabolism differs between cows with subclinical hypocalcemia and their normocalcemic counterparts.     

Short communication: Efficacy of a second intrauterine cephapirin infusion for the treatment of purulent vaginal discharge and endometritis in postpartum dairy cows

The objective of this study was to quantify the efficacy of a second intrauterine cephapirin treatment administered 14 d after the initial one on subsequent reproductive performance of postpartum dairy cows affected by purulent vaginal discharge (PVD) or endometritis (ENDO). In total, 4,140 Holstein cows from 30 commercial herds were enrolled in a randomized clinical trial. At 36 (±7) d in milk, cows were examined using the Metricheck device to diagnose PVD. An endometrial cytology sample was also collected from each cow to perform a leukocyte esterase test for diagnosing ENDO. Diagnosis of PVD and ENDO was done cow-side. Cows diagnosed with PVD or ENDO were assigned to receive 1 of 2 treatments: (1) a single intrauterine cephapirin infusion (500 mg of cephapirin benzathin; Metricure, Merck Animal Health, Kirkland, QC, Canada) at the time of initial examination or (2) a single intrauterine infusion at the time of initial examination and a second one 14 d later. Subsequent reproductive and culling events were collected until 200 d in milk. Statistical analyses were performed using univariable and multivariable mixed logistic regression models. In cows affected by PVD, a second intrauterine cephapirin infusion increased the pregnancy risk at first insemination in comparison with cows that only received one treatment (28.0 vs. 38.8%). In cows affected by ENDO, a second treatment also increased the pregnancy risk at first insemination compared with cows that only received one treatment (30.3 vs. 39.2%). Overall, these results demonstrate that administering a second intrauterine cephapirin infusion 14 d after the initial treatment in postpartum cows affected by PVD or ENDO did improve their subsequent reproductive performance.     

Relationships between endometritis and metabolic state during the transition period in pasture-grazed dairy cows

The primary objective of this study was to identify relationships between endometritis and metabolic state during the calving transition and early lactation periods. A subset of mixed age and breed dairy cows (n = 78) from a seasonal, pasture-grazed herd of 389 cows was examined. The selected cows were grouped as having endometritis at d 42 postpartum or being unaffected by endometritis. Endometritis was defined as >6% (upper quartile) of uterine nucleated cells being polymorphonuclear cells (H-PMN; n = 38); unaffected by endometritis was defined as ≤1% of nucleated cells being polymorphonuclear (L-PMN; n = 40). Milk yield was determined at each milking, and milk composition (fat and protein) was determined at 2-wk intervals. Blood samples collected on d −14, 0 (d of calving), 4, 7, 14, 28, and 42 were analyzed for indicators of energy status (nonesterified fatty acids, glucose, and urea), liver function (albumin, globulin, glutamate dehydrogenase, and aspartate aminotransferase), inflammation (haptoglobin), and mineral status (Ca and Mg). Samples collected weekly from d 21 to 63 or 70 were analyzed for progesterone content. The postpartum anovulatory interval was defined to end on the first day postpartum that plasma progesterone concentration was ≥1 ng/mL. A greater percentage of H-PMN cows failed to ovulate before d 63 or 70 (34%) compared with L-PMN cows (10%), although the proportions of cows ovulating within either polymorphonuclear group was similar through d 56 postpartum. Plasma concentrations of albumin and the albumin:globulin ratio were consistently lower in H-PMN cows. Plasma Mg was lower, whereas glutamate dehydrogenase and aspartate aminotransferase were higher, in H-PMN cows during early lactation compared with L-PMN cows. Circulating metabolites indicative of energy status (nonesterified fatty acids, glucose, and urea) were not different between polymorphonuclear groups. Among 3- to 5-yr-old cows, daily milk yield for the first 42 d after calving was lower for H-PMN cows than for L-PMN cows. Among cows >5 yr old, protein percentage was lower in H-PMN cows compared with L-PMN cows. In summary, endometritis at 42 d postpartum in the herd studied was associated with an increased likelihood of remaining anovulatory. These cows had lower albumin concentrations throughout the calving transition period, perhaps indicating impaired liver function, with lower plasma Mg and evidence of hepatocellular damage in early lactation. Similar profiles of nonesterified fatty acids and glucose indicated that energy status was not a risk factor for endometritis.     

Individual and combined effects of anovulation and cytological endometritis on the reproductive performance of dairy cows

The objective was to evaluate the individual and combined effect of anovulation and cytological endometritis (CTE) on the reproductive performance of dairy cows. A total of 1,569 cows from 3 data sets were used. In data set 1, 403 Holstein cows from 5 dairies in New York were used. In data set 2, 750 Holstein cows from 2 dairies, one in Florida and one in California were used. In data set 3, 416 dairy cows, 165 Holsteins, 36 Jerseys, and 215 Holstein-Jersey crossbreeds from a grazing dairy in Florida were used. Cyclicity and CTE was determined at 35 ± 3 (data set 2) or 49 ± 3 d in milk (data sets 1 and 3). A variable (VarCycCTE) containing all 4 possible permutations between cyclicity (cyclic = Cyc; anovular = Anov) and CTE (present = CTE; absent = Healthy) was created. In the combined data set (sets 1, 2, and 3), pregnancy per artificial insemination (P/AI) diagnosed at 30 to 38 d after first AI was affected by VarCycCTE, with AnovCTE cows having decreased P/AI compared with CycHealthy cows (21.3 vs. 46.7%), whereas AnovHealthy (37.9%) and CycCTE cows (36.0%) had intermediate P/AI. Pregnancy per artificial insemination for the individual data sets and for pregnancy diagnosed at 63 to 74 d after artificial insemination followed a similar pattern. Pregnancy loss was not affected by VarCycCTE. Hazard of pregnancy up to 300 d in milk was affected by VarCycCTE in the combined data sets 1 and 2, with AnovCTE [hazard ratio (HR) = 0.55], AnovHealthy cows (HR = 0.71), and CycCTE (HR = 0.8) having decreased hazard of pregnancy compared with CycHealthy cows. Median days open were 200, 159, 145, and 121 for AnovCTE, AnovHealthy, CycCTE, and CycHealthy, respectively. Hazard of pregnancy for the individual data sets followed a similar pattern. In summary, both anovulation and CTE were negatively associated with reproductive performance and, when combined, they had an additive negative effect.     

Effects of chromium propionate supplementation during the periparturient period and early lactation on metabolism,performance, and cytological endometritis in dairy cows

Multiparous Holstein cows (n = 61) were used to determine the effects of chromium propionate (Cr-Pro) supplementation during the periparturient period and early lactation on metabolism, performance, and the incidence of cytological endometritis (CE). After a 1-wk preliminary period, cows were assigned randomly to 1 of 2 treatments from 21 d before expected calving through 63 d postpartum: (1) control (n = 31) and (2) Cr-Pro (n = 30) administered by daily topdress at a rate of 8 mg/d of Cr. A tendency was detected for increased dry matter intake (DMI) during the prepartum period for cows fed Cr-Pro. Moreover, cows fed Cr-Pro tended to have lower plasma concentrations of nonesterified fatty acids during the prepartum period. However, effects of Cr-Pro supplementation on postpartum DMI and milk yield were not significant. Cows fed Cr-Pro tended to have higher urea N concentrations in milk. An interaction of treatment and day existed during the postpartum period, such that cows fed Cr-Pro had lower plasma glucose concentrations within the first day postpartum compared with controls. Plasma haptoglobin concentration was not affected by treatment during the postpartum period. Blood neutrophil glycogen concentrations were not affected by treatment when sampled at either 7 d postpartum or on one day between 40 and 60 d (48 d ± 0.44 standard error) postpartum. Evaluation of endometrial cytology by low volume lavage at 7 d postpartum (first lavage) and on one day between 40 and 60 d (second lavage) postpartum revealed that cows fed Cr-Pro tended to have a higher percentage of neutrophils at first lavage and decreased incidence of CE as assessed at second lavage. In conclusion, supplementation with Cr-Pro resulted in trends for increased DMI and lower plasma nonesterified fatty acids prepartum. Postpartum production and energy metabolism were not affected by treatment; however, Cr-Pro supplementation tended to affect the postpartum influx of neutrophils into the uterus and decreased the incidence of CE, suggesting positive effects of Cr-Pro supplementation on uterine health.     

Associations of cytological endometritis with energy metabolism and inflammation during the periparturient period and early lactation in dairy cows

Multiparous Holstein cows (n = 108) were used to determine the associations of cytological endometritis (CE) with plasma nonesterified fatty acids (NEFA) and β-hydroxybutyrate (BHBA) as markers of energy metabolism, calculated energy balance (EB), and plasma haptoglobin (Hp) as a marker of inflammation during the periparturient period and early lactation. Evaluation of endometrial cytology by low-volume uterine lavage was conducted on 1 d between 40 and 60 d postcalving. The incidence of CE among cows sampled was 40%. The area under the curve (AUC) was calculated for both NEFA and BHBA using data collected from 3 wk before to 3 wk after parturition. Data for NEFA and BHBA AUC were stratified into prepartum (wk −3 to parturition) and postpartum (parturition to wk +3) for statistical analysis. Prepartum AUC for neither NEFA nor BHBA was associated with subsequent CE; however, cows that subsequently developed CE tended to have higher postpartum AUC for NEFA and had higher postpartum AUC for BHBA. Consistent with the results for plasma NEFA and BHBA, calculated EB during the prepartum period was not different in cows that did or did not develop CE; however, cows with CE had lower EB during the 6-wk postpartum period compared with cows without CE. Analysis of EB by week (wk −3 to −1 before calving and wk +1 to +6 postcalving) indicated that EB in cows with CE was lower at wk +1, +2, and +3 and tended to be lower at wk +6 than cows without CE. Plasma Hp concentrations were analyzed from wk +1 to +8 of lactation; concentrations of Hp were not different during either wk +1 or the entire postpartum period between cows that did or did not develop CE. These results suggest that lower energy status during the first 3 wk postpartum, but not necessarily systemic inflammation, is associated with subsequent development of CE.     

Evaluation of the Metricheck device and the gloved hand for the diagnosis of clinical endometritis in dairy cows

The objective of this study was to compare 2 surrogate vaginal examination methods (i.e., gloved hand and a vaginal device) with vaginoscopy as a reference method for diagnosing clinical endometritis in dairy cows. Holstein-Friesian cows (n = 1,002) in 2 commercial dairy herds in Germany were examined for endometritis at 21 to 27 d in milk (DIM) by using 1 of 3 vaginal diagnostic methods. Vaginal examinations were performed either with a speculum (reference method), a vaginal device (Metricheck, Simcro, New Zealand), or a gloved hand. Vaginal discharge adhering to the diagnostic tool was classified according to a vaginal discharge score ranging from 0 to 3 (where 0 = translucent mucus, 1 = mucus containing flecks of white or off-white pus, 2 = less than 50% white or off-white mucopurulent material, and 3 = greater than 50% white or yellow pus that may be sanguineous). Cows with vaginal discharge scores of 1 to 3 received 500 μg of cloprostenol after examination and again 14 d later (35 to 41 DIM). The prevalence of endometritis in both herds was 40.6 and 40.3%, respectively. With the Metricheck device, significantly more cows were diagnosed as affected with endometritis than by examination with a speculum or a gloved hand (47.5 vs. 36.9 and 36.8%). Binary logistic regression for the risk of conception after first AI as an outcome variable, with vaginal discharge score, diagnostic method, and farm as covariates, revealed a significant effect of degree of endometritis, but not of the diagnostic methods. Survival analyses for the hazard of insemination and pregnancy within 200 DIM, respectively, revealed a significant effect of degree of endometritis, herd, and parity, but not of the diagnostic tool. It can be concluded that any one of the 3 vaginal examination methods can be used interchangeably, without a negative effect on reproductive performance.     

Comparison of ultrasonography and histopathology for the diagnosis of endometritis in Holstein-Friesian cows

Reproductive diseases are a great barrier to dairy production, as they diminish fertility and cause economic loss. In this context, endometritis plays an important role. Different diagnostic techniques exist for endometritis that vary in sensitivity and specificity. This work aimed to evaluate and compare ultrasonographic and histopathological examinations, and combinations thereof, for the diagnosis of endometritis by determining their sensitivity, specificity, and kappa agreement coefficients using endometrial cytology as the gold standard. We developed an adaptive reading score to perform histological examination of the uterus. In this study, 76 Holstein-Friesian cows, 21 to 47 d postpartum, without vaginal catarrh were submitted to gynecological examinations, including ultrasonographic examination, an endometrial cytology examination, and a uterine biopsy. Our results indicate that ultrasonography is a practical and effective diagnostic method, and a combination of the determination of intrauterine fluid and cervical diameter was the most efficient ultrasonographic method, with 50% sensitivity and 88% specificity. Histopathological examination was a useful diagnostic method that provided rich detail on endometrial inflammation, and allowed evaluation of different components of the tissue such as the epithelium, lamina propria, endometrial glands, and blood vessels; this method had 44% sensitivity and 92% specificity for diagnosis of endometritis.     

Subclinical mastitis in dairy cows in major milk-producing areas of Sri Lanka: Prevalence,associated risk factors,and effects on reproduction

Compared with clinical mastitis, the subclinical form of mastitis (SCM) is more common and thought to cause more economic losses to the dairy industry. The current study aimed to investigate the prevalence, risk factors of SCM, and effects on reproduction of dairy cows in major milk-producing areas of Sri Lanka. A total of 1,357 cows of selected farms in 3 regions were examined in the study. California Mastitis Test was conducted for individual cows, and a score of 2 or more for any quarter without any clinical symptoms and abnormalities in milk was considered as positive for SCM. Samples from infected animals were collected and subjected to bacteriological analysis. A pretested questionnaire was used to collect data on individual cows and herds. Risk factors associated with SCM were analyzed using binary logistic regression in generalized linear mixed models. The effect of SCM on calving to conception interval and days from calving to artificial insemination were analyzed by survival analysis using Cox's proportional hazards regression and Kaplan-Meier survival function estimates, respectively. A Poisson regression model was run to determine the effect of SCM on number of artificial inseminations per conception. The prevalence of SCM was 57.5, 11.8, and 45.5% in the regions A, B, and C, respectively. The most common pathogen was Staphylococcus aureus, with 87.1, 56.5, and 92.3% in the regions A, B, and C, respectively. Logistic regression analysis showed that parity, farming system, milking area, region, and herd significantly affect the prevalence of SCM. Subclinical mastitis during the pre-breeding period was associated with 14% increase in the chance of having a greater number of artificial inseminations per conception. Likewise, median days from calving to artificial insemination was longer in cows with SCM compared with normal cows (79 and 64 d, respectively). Therefore, SCM affected the inseminated proportion of studied cows. However, SCM was not associated with the calving to conception interval. The results revealed that the cow factors and milk hygiene play a significant role in the prevalence of SCM.     

Determination of ceftiofur derivatives in serum, endometrial tissue, and lochia in puerperal dairy cows after subcutaneous administration of ceftiofur crystalline free acid

Puerperal uterine infections are often associated with decreased reproductive performance in dairy cows. Routine treatment protocols include the systemic administration of antibiotics. Antibiotic drugs, however, should be administered daily over at least 5 d. The objective of this study was to determine concentrations of ceftiofur derivatives in serum, endometrial tissue, and lochia after subcutaneous administration of ceftiofur crystalline free acid in 6 clinically healthy puerperal dairy cows with normal parturition. Samples were taken immediately before treatment, 2 h after, and then every 24 h over a 7-d period. Concentrations of ceftiofur derivatives were quantified using an HPLC assay. In serum and endometrial tissue, ceftiofur derivatives could be detected above the reported minimum drug concentrations required to inhibit relevant pathogens such as Escherichia coli and Arcanobacterium pyogenes over a 7-d period. Concentrations of desfuroylceftiofuracetamide at 5 d after administration of ceftiofur crystalline free acid were 1.21 ± 0.61 μg/mL in serum, 0.86 ± 0.61 μg/mg in endometrial tissue, and 0.96 ± 1.15 μg/mL in lochia. In lochia, mean concentrations of ceftiofur derivatives also remained above the minimal inhibitory concentration of relevant pathogens, but showed greater variations between cows.     

Prevalence,risk factors,and effects on fertility of cytological endometritis at the time of insemination in Norwegian Red cows

The present study aimed to assess the occurrence of cytological endometritis (CYTO), a nonsymptomatic inflammation of the endometrium, at first artificial insemination (AI) postpartum in Norwegian Red cows. Further, risk factors for CYTO manifestation and its effect on reproductive success and late embryo loss were evaluated. In total 1,648 cows located in 116 herds were included in the study. On mainly spontaneous estrus, endometrial cytology samples were collected using a cytotape technique, and a total of 300 representative epithelial cells and polymorphonuclear neutrophils (PMN) were counted at 400× magnification. Vaginal mucus obtained by Metricheck (Simcro) and body condition score were recorded. Milk samples for progesterone analysis were collected at AI and 21 d later. Pregnancy was diagnosed by rectal palpation or analysis of pregnancy-associated glycoproteins. Based on the constructions of a receiver operator characteristics curve, the cut-off level for PMN defined as CYTO was set to 3.0%, representing the level at which the PMN occurrence affected pregnancy outcome, with the highest summation of sensitivity (32.4%) and specificity (74.9%). Three logistic models with herd included as random factor were constructed. The outcome for the first model was the likelihood for CYTO based on the endometrial samples, in the second model pregnancy to first AI, and in the third model embryo loss. The proportion of CYTO was 28.0% (461/1,648). The average interval in days to first AI was 71.7 d (standard error ± 0.7) and the overall pregnancy incidence to first AI was 59.8% (866/1,449). The likelihood for CYTO at first AI was associated with AI personnel, calving to first AI interval, vaginal mucus characteristics, amount of red blood cells in sample, season, and barn type. Pregnancy to first AI was lower in CYTO-positive cows (odds ratio = 1.51, confidence interval = 1.17–1.94). Other factors affecting pregnancy to first AI were AI personnel, test day milk yield, barn type, and obstetrical conditions or fertility treatments before first AI. The proportion of late embryo loss and abortion was 8.6% (82/948) and 2.8% (24/866), respectively. Late embryo loss was associated with treatment against fertility disorders before first AI, but not associated with CYTO. Overall, our results suggest that even if Norwegian Red cows show a fairly high prevalence of CYTO in the endometrium at first AI, it does not seem to have a major effect on the reproductive performance. The Norwegian Red breeding program has emphasized fertility and health for decades, and a genetically advantageous uterine immunology might be one of the preserved mechanisms.     

Effects of feeding dry glycerin to early postpartum Holstein dairy cows on lactational performance and metabolic profiles

Effects of feeding a dry glycerin product (minimal 65% of food grade glycerol, dry powder) to 39 multiparous Holstein dairy cows (19 control and 20 glycerin-supplemented; lactation number = 2.2 ± 1.3 SD) on feed intake, milk yield and composition, and blood metabolic profiles were investigated. Dry glycerin was fed at 250 g/d as a top dressing (corresponding to 162.5 g of glycerol/d) to the common lactating total mixed ration from parturition to 21 d postpartum. Individual milk was sampled from 2 consecutive milkings weekly and analyzed for components. Blood was sampled from the coccygeal vein at 4, 7, 14, and 21 (±0.92, pooled SD) d in milk and analyzed for urea nitrogen, glucose, insulin, nonesterified fatty acids, and β-hydroxybutyrate. Urine was tested for the acetoacetate level weekly by using Ketostix. Average feed intake, milk yield and components, blood metabolites, and serum insulin concentrations were not affected by dry glycerin supplementation. Glycerin-supplemented cows experienced a more positive energy status (higher concentrations of plasma glucose, lower concentrations of plasma β-hydroxybutyrate, and lower concentrations of urine ketones), which was observed during the second week of lactation, suggesting that energy availability may have been improved. This glucogenic effect of dry glycerin did not result in an increase in feed intake or milk yield during the first 3 wk of lactation, likely because of the relatively less negative energy status of cows transitioning into lactation. The tendency toward higher milk yield for glycerin-supplemented cows during wk 6 of lactation (52 vs. 46 kg/d) after the supplementation period (dry glycerin was terminated at wk 3 of lactation) suggested a potential benefit of dry glycerin on subsequent milk production, perhaps through changes in metabolism, which requires further investigation.     

Association of immediate postpartum plasma calcium concentration with early-lactation clinical diseases,culling, reproduction,and milk production in Holstein cows

The objective of this study was to evaluate the association of postpartum plasma Ca concentration with early-lactation disease outcomes, culling within 60 d in milk, pregnancy to first service, and milk production. A total of 1,453 cows from 5 commercial dairy farms in New York State were enrolled in a prospective cohort study from February to November 2015. Blood samples were collected within 12 h of parturition, and plasma was submitted to a diagnostic laboratory for total Ca measurement. Early-lactation disease, reproductive performance, and milk production from Dairy Herd Improvement Association (DHIA) test-day data were compiled from each farm's management software. Multivariable Poisson regression models were built to evaluate the association of plasma Ca with the risks of retained placenta (RP), metritis, displaced abomasum (DA), clinical mastitis, culling within 60 d in milk, and pregnancy to first service. Repeated-measures ANOVA were used to evaluate the association of Ca at parturition with milk production across the first 9 DHIA tests. Herd was considered a random effect in all models. Primiparous cows were modeled separately from multiparous cows if differential responses were observed. Calcium was not associated with the risk of RP, metritis, clinical mastitis, or pregnancy to first service in primiparous or multiparous cows. For multiparous cows only, higher Ca concentration tended to be associated with increased culling within the first 60 d in milk. Multiparous cows with Ca ≤1.85 mmol/L had an increased risk of being diagnosed with a DA compared with cows with Ca >1.85 mmol/L. For the milk production models, Ca was not associated with the amount of milk produced within the first 9 DHIA tests in primiparous cows; however, multiparous cows with Ca ≤1.95 mmol/L produced, on average, 1.1 kg more milk per day across the 9 DHIA tests than their multiparous counterparts with Ca >1.95 mmol/L. Our results indicate that plasma Ca concentration measured within 12 h of parturition is a poor predictor of early-lactation health outcomes. Reduced Ca concentration in the immediate postpartum period was associated with higher milk production in multiparous cows. From these results, we caution that studies attempting to categorize subclinical hypocalcemia based on a single sample in the immediate postpartum period could misclassify the disorder.