Effects of source of supplementary trace minerals in pre- and postpartum diets on reproductive biology and performance in dairy cows.

Our objectives were to evaluate the effects of complete replacement of inorganic salts of trace minerals (STM) with organic trace minerals (OTM) in both pre- and postpartum diets on ovarian dynamics, estrous behavior measured by sensors, preimplantation conceptus development, and reproductive performance in dairy cows. Pregnant cows and heifers (n = 273) were blocked by parity and body condition score and randomly assigned to either STM or OTM diets at 45 ± 3 d before their expected calving. Pre- and postpartum diets were formulated to meet 100% of recommended levels of each trace mineral in both treatments, taking into consideration both basal and supplemental levels. The final target concentrations of Co, Cu, Mn, Se, and Zn were, respectively, 0.25, 13.7, 40.0, 0.3, and 40.0 mg/kg in the prepartum diet, and 0.25, 15.7, 40.0, 0.3, and 63.0 mg/kg in the postpartum diet. The STM group was supplemented with Co, Cu, Mn, and Zn sulfates and sodium selenite, while the OTM group was supplemented with Co, Cu, Mn, and Zn proteinates and selenized yeast. Treatments continued until 156 d in milk (DIM) and were assigned to individual cows using automatic feeding gates. Starting at 21 DIM, ultrasonography examinations of the ovaries were performed weekly to determine the presence of a corpus luteum and postpartum resumption of ovarian cyclicity. Cows were presynchronized with 2 injections of PGF2α at 42 and 56 DIM. Estrous behavior was monitored using electronic activity tags that indirectly measured walking activity. Cows detected in estrus after the second PGF2α were inseminated, and those not detected in estrus by 67 DIM were enrolled in a synchronization program. Cows that returned to estrus after artificial insemination (AI) were reinseminated. Pregnancy diagnosis was performed 33 d after AI, and nonpregnant cows were resynchronized. Transcript expression of interferon-stimulated genes in peripheral blood leukocytes was performed in a subgroup of cows (STM, n = 67; OTM, n = 73) on d 19 after AI. A different subgroup of cows (28 STM, 29 OTM) received uterine flushing 15 d after AI for recovery of conceptuses and uterine fluid for analyses of transcriptomics and metabolomics, respectively. In addition, dominant follicle diameter, luteal size and blood flow, and concentration of progesterone in plasma were measured on d 0, 7, and 15 relative to AI. After flushing, PGF2α was given and the dominant follicle was aspirated 2 d later to measure the concentration of trace minerals by mass spectrometry. Estrous behavior, size of the dominant follicle and corpus luteum, concentration of progesterone, time to pregnancy, and proportion of cows pregnant by 100 d of the breeding period did not differ between treatments. A greater proportion of cows supplemented with OTM had a corpus luteum detected before presynchronization (64.3 vs. 75.2%), and primiparous cows supplemented with OTM tended to resume cyclicity earlier than their STM counterparts. Cows supplemented with OTM had a greater concentration of Cu in follicular fluid than cows supplemented with STM (0.89 vs. 0.77 µg/mL, respectively). In pregnant multiparous cows, expression of receptor transporter protein 4 in peripheral blood leukocytes was 42% greater in the OTM group. Conceptuses of the 2 treatments had 589 differentially expressed transcripts, with many indicating advanced conceptus elongation and greater transcript expression of selenoproteins in the OTM group. In pregnant cows, 24 metabolites were more abundant in the uterine fluid of OTM, including spermidine, sucrose, and cholesterol. In conclusion, replacing STM with OTM caused modest improvements to resumption of ovarian cyclicity and important changes in preimplantation conceptus development, but it did not alter conception risk and pregnancy rate.

Bayesian estimation of sensitivity and specificity of a milk pregnancy-associated glycoprotein ELISA test for pregnancy diagnosis between 23 and 27 days after insemination in Holstein dairy cows.

Pregnancy diagnosis using pregnancy-associated glycoprotein (PAG) ELISA technology in blood or milk samples is validated from 28 d after insemination in dairy cows. The objective of this study was to estimate the sensitivity (Se) and specificity (Sp) of a commercial milk PAG-based ELISA in Holstein dairy cows between 23 and 27 d after insemination. Milk samples (n = 268) from 257 Holstein dairy cows 23 to 27 d after AI were submitted for PAG ELISA testing. Pregnancy status was confirmed by either a second milk PAG ELISA test conducted between 28 and 50 d after insemination (n = 200) or transrectal ultrasonography performed between 28 and 59 d after insemination (n = 68). A Bayesian latent class model was used to compare the paired results from the test at 23 to 27 d after AI test to the reference test. The latent class model typically used for comparing 2 or more imperfect tests was extended to include the possibility of pregnancy loss between the 23 to 27 d test and the reference test. Informative priors for the probability of pregnancy loss, and for the Se and Sp of the PAG and ultrasonography reference tests were obtained from the scientific literature. Estimated median Se and Sp of the PAG ELISA test conducted between 23 and 27 d after AI were 0.98 (95% credible interval 0.93 to 1.0) and 0.98 (0.89 to 1.0), respectively, when using a standardized corrected optical density threshold of 0.15. Although the accuracy of the test under investigation was excellent, more data will be needed to confirm the optimal diagnostic cut point for PAG in milk for early pregnancy diagnosis in this time window. The optimal timing of pregnancy diagnosis will depend on herd-specific logistics and the action to be taken to re-inseminate nonpregnant cows.

Effects of replacing inorganic salts of trace minerals with organic trace minerals in pre- and postpartum diets on feeding behavior, rumen fermentation, and performance of dairy cows.

Our objectives were to evaluate the effects of complete replacement of supplementary inorganic salts of trace minerals (STM) by organic trace minerals (OTM) in both pre- and postpartum diets on feeding behavior, ruminal fermentation, rumination activity, energy metabolism, and lactation performance in dairy cows. Pregnant cows and heifers (n = 273) were blocked by parity and body condition score and randomly assigned to either STM or OTM diets at 45 ± 3 d before their expected calving date. Both groups received the same diet, except for the source of trace minerals (TM). The STM group was supplemented with Co, Cu, Mn, and Zn sulfates and Na selenite, whereas the OTM group was supplemented with Co, Cu, Mn, and Zn proteinates and selenized yeast. Treatments continued until 156 days in milk and pre- and postpartum diets were formulated to meet 100% of recommended levels of each TM in both treatments, taking into consideration both basal and supplemental levels. Automatic feed bins were used to assign treatments to individual cows and to measure feed intake and feeding behavior. Rumination activity was monitored by sensors attached to a collar from wk -3 to 3 relative to calving. Blood metabolites were evaluated on d -21, -10, -3, 0, 3, 7, 10, 14, 23, and 65 relative to calving. Ruminal fluid samples were collected using an ororuminal sampling device on d -21, 23, and 65 relative to calving, for measurement of ruminal pH and concentration of volatile fatty acids. Cows were milked twice a day and milk components were measured monthly. Cows supplemented with OTM tended to have longer daily feeding time (188 vs. 197 min/d), and greater dry matter intake (DMI; 12.9 vs. 13.3 kg), and had a more positive energy balance (3.6 vs. 4.2 Mcal/d) and shorter rumination time per kg of dry matter (DM; 40.1 vs. 37.5 min/kg of DM) than cows supplemented with STM during the prepartum period. In the postpartum period, OTM increased DMI in multiparous cows (24.1 vs. 24.7 kg/d) but not in primiparous cows (19.1 vs. 18.7 kg/d). The difference in DMI of multiparous cows was more evident in the first 5 wk of lactation, when it averaged 1 kg/d. Milk yield was not affected by treatment in multiparous cows (44.1 vs. 44.2 kg/d); however, primiparous cows supplemented with OTM had lesser yields than primiparous cows supplemented with STM (31.9 vs. 29.8 kg/d). Cows supplemented with OTM had a greater percentage of protein in milk (3.11 vs. 3.17%), reduced concentration of nonesterified fatty acids in serum (0.45 vs. 0.40 mmol/L), and rumination activity (30.1 vs. 27.8 min/kg of DM) than cows supplemented with STM. At the end of the transition period, cows supplemented with OTM had reduced molar proportion of acetate, reduced pH, and tended to have a greater concentration of total volatile fatty acids in ruminal fluid. In conclusion, complete replacement of STM by OTM caused modest changes in rumen fermentation, feeding behavior, energy metabolism, and performance of dairy cows, improving postpartum DMI in multiparous cows and reducing circulating levels of nonesterified fatty acids. The pre-absorptive effects of TM source and the parity specific responses on performance warrant further research.

Effects of parity, blood progesterone, and non-steroidal anti-inflammatory treatment on the dynamics of the uterine microbiota of healthy postpartum dairy cows.

This study evaluated the effects of treatment with meloxicam (a non-steroidal anti-inflammatory drug), parity, and blood progesterone concentration on the dynamics of the uterine microbiota of 16 clinically healthy postpartum dairy cows. Seven primiparous and 9 multiparous postpartum Holstein cows either received meloxicam (0.5 mg/kg SC, n = 7 cows) once daily for 4 days (10 to 13 days in milk (DIM)) or were untreated (n = 9 cows). Endometrial cytology samples were collected by cytobrush at 10, 21, and 35 DIM, from which the microbiota analysis was conducted using 16S rRNA gene sequence analysis. A radioimmunoassay was used to measure progesterone concentration in blood serum samples at 35 DIM and cows were classified as ˃ 1 ng/mL (n = 10) or ≤ 1 ng/mL (n = 6). Alpha diversity for bacterial genera (Chao1, Shannon-Weiner, and Camargo's evenness indices) were not affected by DIM, meloxicam treatment, parity, or progesterone category. For beta diversity (genera level), principal coordinate analysis (Bray-Curtis) showed differences in microbiota between parity groups. At the phylum level, the relative abundance of Actinobacteria was greater in primiparous than multiparous cows. At the genus level, there was lesser relative abundance of Bifidobacterium, Lactobacillus, Neisseriaceae, Paracoccus, Staphylococcus, and Streptococcus and greater relative abundance of Bacillus and Fusobacterium in primiparous than multiparous cows. Bray-Curtis dissimilarity did not differ by DIM at sampling, meloxicam treatment, or progesterone category at 35 DIM. In conclusion, uterine bacterial composition was not different at 10, 21, or 35 DIM, and meloxicam treatment or progesterone category did not affect the uterine microbiota in clinically healthy postpartum dairy cows. Primiparous cows presented a different composition of uterine bacteria than multiparous cows. The differences in microbiota associated with parity might be attributable to changes that occur consequent to the first calving, but this hypothesis should be investigated further.

Effect of anti-inflammatory treatment on systemic inflammation, immune function, and endometrial health in postpartum dairy cows.

Systemic inflammation (SI) is increasingly studied in several species because it may be central in many metabolic disturbances and be a risk factor for clinical disease. This proof-of-concept study evaluated the effects of the anti-inflammatory drug meloxicam on markers of SI and energy metabolism, polymorphonuclear neutrophil (PMN) function, and endometritis in clinically healthy postpartum dairy cows. Cows received meloxicam (0.5 mg/kg of body weight; n = 20) once daily for 4 days (10-13 days postpartum) or were untreated (n = 22). Blood samples were collected -7, 1, 3, 5, 7, 10, 11, 12, 13, 14, 18, 21, 28, and 35 days relative to calving to measure serum concentrations of metabolic and inflammatory markers. Function of peripheral blood PMN were evaluated at 5, 10, 14, and 21, and proportion of PMN in endometrial cytology were performed at 5, 10, 14, 21, 28 and 35 days postpartum. Meloxicam decreased serum haptoglobin from the second until the last day of treatment, and improved indicators of energy metabolism (lesser ß-hydroxybutyrate and greater insulin-like growth factor-1 during treatment, and greater glucose at the end of treatment than control cows). This improved PMN function at 14 days postpartum, but the endometrial inflammatory status was not affected.

Comparison of cow-side diagnostic techniques for subclinical endometritis in dairy cows.

A limited number of cow-side diagnostic techniques exist for the diagnosis of subclinical endometritis (SCE) in dairy cattle. The objectives of this study were to compare results of endometrial cytology from samples collected by cytobrush (CB) and low-volume lavage (LVL) and to assess leukocyte esterase (LE) test strips and Brix refractometry as surrogate cow-side tests for SCE. Two samples were consecutively collected from 248 Holstein cows between 29 and 35 days postpartum, using CB and LVL techniques. Each sample was analyzed using cytology with a cut-point of ≥5% polymorphonuclear (PMN) cells, LE strips using cut-points of ≥1 and ≥ 2, and a Brix refractometer. Each diagnostic technique was compared intra-sample using the respective cytology as a gold standard and inter-sample using CB samples as the referent. The concordance correlation coefficient (CCC, ρc) for PMN% between CB and LVL was ρc = 0.59 [95% confidence interval (CI): 0.50 to 0.67] and the Kappa (κ) for agreement was κ = 0.35 [sensitivity (Se) = 0.88, specificity (Sp) = 0.45]. The optimal cut-point of LE ≥ 2 resulted in moderate agreement between CB and LVL samples, κ = 0.56 (Se = 0.89, Sp = 0.65). Agreement between LE and cytology using CB (κ = 0.49; Se = 0.89, Sp = 0.57) and LVL (κ = 0.44; Se = 0.77, Sp = 0.67) were similar. The correlation between Brix values from CB and LVL was ρc = 0.12 (CI -0.01 to 0.26). The correlation between CB cytology and Brix was ρc = 0.33 (CI 0.20 to 0.45) but ρc = -0.07 (CI -0.21 to 0.06) between LVL cytology and Brix. While LE strips with a cut-point of LE ≥ 2 had moderate agreement with cytology, Brix refractometry had poor performance for the diagnosis of SCE. Samples taken by CB and LVL produced comparable cow-side diagnostic results and either is a viable method for the diagnosis of SCE.

Exposure to an unpredictable and competitive social environment affects behavior and health of transition dairy cows.

Social factors are important determinants of disease in humans and and laboratory animals, but less research has been done using farm animals. The objective of this study was to determine if an unpredictable and competitive social environment affects behavior and health during the transition period when dairy cows are at high risk of disease. Five weeks before calving, 64 cows were assigned to a predictable and noncompetitive social environment (predictable) or an unpredictable and competitive social environment (unpredictable) using 8 groups of 4 animals per treatment. Each group consisted of 3 multiparous and 1 primiparous cow. At first enrollment (baseline; 5 wk before calving), all groups had access to 4 electronic feed bins. At 4 wk before calving, cows in the predictable groups were given access to 6 feed bins, and cows in the unpredictable groups were moved into a new pen with 4 resident cows each trained to consume feed from one bin. Each cow in the unpredictable group was then provided access to only 1 of the 4 feed bins which they shared with 1 resident cow (resulting in 2 cows/bin), creating a competitive feeding environment. To create an unpredictable environment, access to morning feed was delayed 0, 1, 2, or 3 h every other day. On alternate days, the cows in unpredictable groups were assigned to feed from a new feed bin (and thus had to compete with a new resident partner). Feeding and social behavior were collected electronically from the feed bins. Blood was sampled at baseline (wk -5), wk -2, wk -1, and wk +1 relative to calving to measure inflammatory (haptoglobin and tumor necrosis factor-α) and metabolic (nonesterified fatty acids, ß-hydroxybutyrate, calcium, and glucose) biomarkers. Uterine cytology was performed 3 to 5 wk after calving to diagnose cytological endometritis. Data were analyzed using mixed models including baseline data as a covariate, week as a repeated measure, treatment as a main effect, and a treatment by week interaction. The probability of cytological endometritis at the group level was analyzed using Mann-Whitney U tests. Parity was included in separate models to determine any parity × treatment interactions. Cows from both treatments consumed the same amount of feed, but cows in the unpredictable group spent less time feeding and had a higher rate of feed intake. Cows in the unpredictable groups also visited the feed bins less often, consumed more feed during each visit, and were involved in more social replacements at the feed bin compared with predictable groups. Cows in the unpredictable groups had higher serum concentrations of nonesterified fatty acids and tumor necrosis factor-α, but lower ß-hydroxybutyrate compared with predictable groups. Multiparous cows in unpredictable groups were more likely to be diagnosed with cytological endometritis after calving compared with cows in the predictable groups, but primiparous cows in unpredictable groups showed a tendency for the opposite response. These results suggest that an unpredictable and competitive social environment before calving causes changes in feeding and social behavior, some physiological indicators of metabolism and inflammation, and increases the risk of uterine disease in multiparous cows after calving.

Estrous detection intensity and accuracy and optimal timing of insemination with automated activity monitors for dairy cows.

The objectives of this observational study were to assess the ability of automated activity monitoring (AAM) to detect estrus for first insemination, the accuracy of detection, and the optimum interval from the estrus alert from the AAM system to insemination. Four commercial farms using 1 of 2 commercial AAM systems were studied over 1 yr. Cows were inseminated between 55 and 80 d in milk (DIM) based on AAM only, then by a combination of AAM and timed artificial insemination (AI). Blood progesterone was measured in 1,014 cows at wk 5, 7, and 9 postpartum; purulent vaginal discharge (PVD) was assessed at wk 5; and lameness and BCS at wk 7. Overall, AAM detected 83% of cows in estrus by 80 DIM. Cows that had 3 serum progesterone <1 ng/mL, had PVD, or were both lame and had BCS ≤2.5 has lesser odds of being detected in estrus by 80 DIM (62, 68, and 53%, respectively). Blood samples were collected on the day of 445 AI based on AAM and 323 timed AI. The proportion of cows not in estrus (progesterone >1 ng/mL) on the day of AI was similar between AAM (4 ± 1.8%) and timed AI (3 ± 1.2%). Managers elected, based on subjective criteria, not to inseminate 17% of cows for which an AAM estrus alert was issued, of which 43% were not in estrus. Activity data were extracted from AAM software for 1,399 AI. Onset of estrus was calculated using the same or similar data processing criteria as the AAM system. Producers recorded the time of AI. The interval from onset of estrus to AI was categorized as 0 to 8, 8 to 16, or 16 to 24 h. We found no effect of AAM system on the probability of pregnancy per AI, but noted an interaction of interval with parity. For multiparous cows, the probability of pregnancy per AI was 31%, which did not differ with the interval to AI. For primiparous cows, the odds of pregnancy were greater if AI occurred 0 to 8 h (49%) than 8 to 16 (36%) or 16 to 24 h (31%) after the estrus alert from the AAM. Automated activity monitoring can detect estrus for first AI in just over the length of 1 estrous cycle for over 80% of cows, but the remainder would likely require intervention for timely insemination. For multiparous cows, performing AI based on AAM once per day would not affect pregnancy per AI, but for primiparous cows AI within 8 h of the onset of estrus may be advantageous.

Validation of commercial luminometry swabs for total bacteria and coliform counts in colostrum-feeding equipment.

A sufficient quantity and quality of colostrum must be fed quickly to the newborn calf while minimizing bacterial contamination. Adenosine triphosphate bioluminescence swabs offer a potential rapid on-farm alternative to assess bacterial contamination of colostrum. The objective of this study was to validate the Hygiena (Camarillo, CA) AquaSnap Total (AS), SuperSnap (SS), PRO-Clean (PC), and MicroSnap Coliform (MS) swabs as well as visual hygiene assessment for detection of elevated bacterial counts in or on colostrum-feeding equipment. From April to October 2016, 18 esophageal tube feeders, 49 nipple bottles, and 6 pails from 52 dairy farms in Ontario were evaluated for cleanliness. Following visual hygiene assessment, sterile physiological saline (15 mL) was poured into each piece of equipment, mixed for 2 min to ensure total surface coverage, and poured into a sterile collection container through the feeding end. The fluid was split into equal aliquots, with one being evaluated by conventional culture and the other evaluated using the luminometry swabs. Nonparametric receiver operator curves were used to compare the test performance of the luminescence reading (relative light units; RLU) from each type of swab to conventional bacterial culture. The area under the curve comparing the AS swab to total bacterial count (cut point >100,000 cfu/mL) was 0.89, and using a cut point of 631 RLU correctly classified 84% of samples with a sensitivity of 88% and a specificity of 77%. The area under the curve comparing the MS swab to total coliform count (cut point >10,000 cfu/mL) was 0.85, and using a cut point of 44 RLU correctly classified 89% of samples with a sensitivity of 83% and a specificity of 90%. Visual hygiene assessment, PC and SS swabs were not reliable indicators for feeding equipment cleanliness. The results suggest that the AS and MS swabs can be used as an alternative to traditional laboratory bacterial counts to evaluate cleanliness of colostrum-feeding equipment.

Reproductive tract inflammatory disease in postpartum dairy cows.

Up to half of dairy cows are affected by at least one of metritis, purulent vaginal discharge, endometritis or cervicitis in the postpartum period. These conditions result from inadequate immune response to bacterial infection (failure to clear pathogenic bacteria from the uterus) or persistent inflammation that impairs rather than enhances reproductive function. The degree of mobilization of fat and how effectively it is used as a metabolic fuel is well recognized as a risk factor for metabolic and infectious disease. Release of non-esterified fatty acids has direct effects on liver and immune function but also produces pro-inflammatory cytokines (tumor necrosis factor α and interleukin-6), which contribute to systemic inflammation and to insulin resistance. Therefore, reproductive tract inflammatory disease may be a function of both local and systemic inflammatory stimuli and regulation as well as regulation of fat metabolism. Better understanding of variables associated with insulin resistance and inflammatory regulation in the liver and adipose tissue may lead to improvement of reproductive tract health. This paper reviews factors that may contribute to postpartum reproductive tract inflammatory diseases in dairy cows and their inter-relationships, impacts and treatment.

Definitions and diagnosis of postpartum endometritis in dairy cows.

The objectives of this observational study were to determine and compare diagnostic criteria for postpartum endometritis in dairy cows. Data generated from 1,044 Holstein cows (6 herds) enrolled in a randomized clinical trial were used. Cows were examined for endometritis at 35±3 d (exam 1) and 56±3 d (exam 2) after parturition, using endometrial cytology (cytobrush technique), vaginal discharge scoring (Metricheck device; Simcrotech, Hamilton, New Zealand), and cervical diameter measurement (transrectal palpation). Reproductive data were recorded until 200 d after parturition. Diagnostic criteria for cytological and clinical endometritis were determined based on detrimental effect on subsequent reproductive performance, using logistic regression and Cox proportional hazard models accounting for the effect of herd clustering. Comparison of diagnostic criteria was performed using endometrial cytology as reference test or by quantifying the agreement between diagnostic approaches. At exam 1, diagnostic criteria were ≥6% polymorphonuclear cells and mucopurulent or worse (purulent or foul) vaginal discharge for cytological and clinical endometritis, respectively. At exam 2, diagnostic criteria were ≥4% polymorphonuclear cells and mucopurulent or worse vaginal discharge for cytological and clinical endometritis, respectively. Cows were classified as having cytological endometritis only, clinical endometritis only, or both cytological and clinical endometritis. Prevalence at exam 1 was 13.5, 9.4, and 5.8% for cytological endometritis only, clinical endometritis only, and both cytological and clinical endometritis, respectively. The detrimental effects of cytological and clinical endometritis on reproductive performance were additive. Among cows with clinical endometritis, only 38 and 36% had cytological endometritis at exam 1 and exam 2, respectively. Combination of diagnostic criteria improved neither the accuracy for predicting cytological endometritis nor the agreement between cytological and clinical endometritis. Overall, these results suggested that cytological and clinical endometritis may represent different manifestations of reproductive tract disease. They also suggested that use of the terminology clinical endometritis may not be accurate and that purulent vaginal discharge may be more descriptive.

Synchronization of estrus and pregnancy risk in anestrous dairy cows after treatment with a progesterone-releasing intravaginal device.

Lactating Holstein cows (located in 4 dairy herds) that had failed to display estrus as defined by increased pedometer activity by 63 +/- 3 d in milk, were enrolled to investigate the effect of a progesterone-releasing intravaginal device (PRID, n = 268) relative to a placebo intravaginal device (PID, control, n = 266) on days from device removal to artificial insemination (AI), the probability of pregnancy at first AI, and days from device removal to pregnancy. Cows were assigned randomly to receive a PRID or PID for 7 d and an injection of PGF2alpha at device removal. Upon device removal, a vaginitis score was assigned and AI occurred at observed estrus. Cows failing to display estrus within 14 d of device removal were subjected to a subsequent reproductive exam and were treated with PGF2alpha. Two percent of PRID-treated cows and 11% of control cows displayed estrus during the 7-d exposure period. Among the remaining cows, 93% of the devices were present at the scheduled removal. Cows treated with the PRID were 60% less likely to have purulent debris on the device than control cows. Vaginal reaction, however, was not associated with any of the reproductive outcomes. Investigation of the reproductive outcomes revealed a treatment x parity interaction. Progesterone-treated primiparous cows were inseminated 17 d earlier, with no significant change in the probability of pregnancy at first AI (30.3 vs. 42.0%), and no difference in median time from device removal to pregnancy (52 vs. 53 d) relative to control primiparous cows. Conversely, PRID-treated multiparous cows were inseminated 8 d earlier, with no change in probability of pregnancy at first AI (24.6 vs. 18.8%); however, median time from device removal to pregnancy was reduced by 20 d (67 vs. 87 d). These results support the efficacy of a PRID to induce estrus in previously anestrous cows. The reason, however, for the variable response between primiparous and multiparous cows was not clear.

The effect of a progesterone releasing intravaginal device (PRID) on pregnancy risk to fixed-time insemination following diagnosis of non-pregnancy in dairy cows.

The objective was to compare the probability of pregnancy after fixed-time insemination in cows diagnosed as non-pregnant and re-inseminated following the Ovsynch protocol, with or without exogenous progesterone. Cows (n=415) used in this study originated from 25 farms. Upon diagnosis of non-pregnancy between 30 and 60 days after AI, cows were randomly assigned to receive either a progesterone releasing intravaginal device (PRID; n=208) or a placebo intravaginal device (PID; n=207). All cows received GnRH at enrollment (Day 0), PGF(2alpha) concurrent with intravaginal device removal 7 days later, GnRH on Day 9 and fixed-time insemination 16h later (Day 10). Cows observed in estrus prior to Day 7, had the device removed and were inseminated. Ovaries were examined by transrectal palpation at the time of enrollment and the prominent structures were assessed and recorded. Body condition score, lameness status, interval from previous insemination, and times bred at enrollment were recorded. At intravaginal device removal, the occurrence and intensity of vaginitis was determined according to the amount of debris on the device. Overall, the intravaginal device retention rate was 91%. A total of 5.2% of PID-treated cows and 2.9% of PRID-treated cows were detected in estrus within the 7 days treatment period. Pregnancy status was diagnosed between 30 and 56 days after insemination and all cows were followed for a minimum of 150 days after enrollment. Approximately 28% of cows had evidence of mild vaginitis in response to the intravaginal device, whereas 6% of cows had copious debris associated with the intravaginal device at removal. The probability of pregnancy after fixed-time insemination was 43.8% versus 34.9% in PRID-treated versus PID-treated animals. Exogenous progesterone provided through an intravaginal device to non-pregnant cows that had not displayed estrus improved the probability of pregnancy after fixed-time AI.

Prevalence and risk factors for postpartum anovulatory condition in dairy cows.

The objectives of this research were to determine the prevalence of the anovulatory condition within a temperate region of North America and identify cow-level and herd-level risk factors for this condition. A total of 1,341 cows from 18 herds were classified as cycling or anovular based on skim milk progesterone concentration determined at 46 and 60 +/- 7 d in milk. Calving history, periparturient disease incidence, body condition score, milk ketone concentration in the first 2 wk of lactation, and first 305-d mature-equivalent milk projections were recorded. Reproductive and culling information was retrieved monthly from the Dairy Herd Improvement Association. The cow-level prevalence of anovulation was 19.5%, with a herd-specific range from 5 to 45%. Accounting for the effect of clustering at the herd level, cows experiencing a difficult calving, cows with twin calvings, displaced abomasum, and cows with subclinical ketosis in the first week after calving were at greater risk for diagnosis of anovulation. Anovular cows within herds using ovulation synchronization programs were inseminated at the same time postpartum with a 6-percentage point reduction in the probability of pregnancy relative to cycling herdmates (29.7 vs. 35.9%, respectively), whereas anovular cows in herds breeding based on observed estrus were inseminated 8 d later and suffered a 10-percentage point reduction in the probability of pregnancy at first insemination (20.3 vs. 30.5). Time to pregnancy was delayed in anovular cows by 30 d (156 vs. 126 d). Using survival analysis, the impact of anovulation decreased with time. The daily probability of pregnancy (hazard ratio) was similar to cycling cows by 165 d in milk. The results underline the important associations of peripartum health with reproductive function and performance.

Short communication: presynchronization using a single injection of PGF2alpha before synchronized ovulation and first timed artificial insemination in dairy cows.

Conception to synchronized ovulation (Ovsynch) using injections of GnRH and PGF2alpha and timed artificial insemination has been shown to be maximized when the program is initiated 5 to 12 d after estrus. The objective of this double-blinded field trial was to assess the effect of one injection of PGF2alpha, 10 d before the Ovsynch program, on the probability of pregnancy at first insemination in lactating dairy cows. The hypothesis was that cows that underwent luteolysis in response to PGF2alpha would be between 5 and 8 d postestrus at the start of Ovsynch. In five commercial dairy herds in Ontario, Canada, at 52 +/- 12 d in milk, 506 cows were assigned at random to receive either one i.m. injection of 500 microg of cloprostenol or saline. Ten days later, all cows received 100 microg of GnRH i.m., followed in 7 d by 500 microg of cloprostenol i.m. and 100 microg of GnRH i.m. 48 h later. All cows were artificially inseminated 0 to 20 h after the second injection of GnRH, without regard to detection of estrus. Pregnancy was diagnosed by transrectal palpation at least 35 d after insemination. The probability of pregnancy after first insemination was modeled with logistic regression, accounting for the correlation of cows with herd and the effect of season of calving. There was no difference in pregnancy risk between cows that received PGF2alpha presynchronization and controls (37.3 and 36.6%, respectively; odds ratio = 1.03, 95% confidence interval, 0.88 to 1.20). Parity and days in milk at insemination were not significant covariates.

Measures of estrus detection and pregnancy in dairy cows after administration of gonadotropin-releasing hormone within an estrus synchronization program based on prostaglandin F2 alpha.

The objective of this study was to assess the use of GnRH in a controlled breeding program (7 d prior to the second of two injections of PGF2 alpha, 14 d apart) as a means to improve the expression and detection of estrus, first service conception rate, and overall pregnancy rate of lactating dairy cows. On 17 farms, 348 cows were assigned randomly to either of two breeding programs prior to first insemination. Cows in both programs received PGF2 alpha approximately 2 wk prior to the end of a herd-specific voluntary waiting period for breeding. One group received GnRH 1 wk later, 7 d prior to the second of two injections of PGF2 alpha. Control cows received saline and a second injection of PGF2 alpha at corresponding times. Cows were observed for 7 d and were bred by artificial insemination following detection of estrus. There were no differences between programs in estrus detection rate, observed signs of estrus, conception rate, days to first service, or interval from calving to conception. The means and standard deviations of the interval from PGF2 alpha to estrus were not different between programs. Administration of GnRH 1 wk prior to PGF2 alpha did not alter the expression of estrus or fertility in lactating dairy cows. In this study population, no advantage was found for the addition of GnRH to a controlled breeding program that was based on two administrations of PGF2 alpha at a 14-d interval.