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The objective of this series of studies was to investigate the effects of inflammatory diseases occurring before breeding on the developmental biology and reproductive responses in dairy cows. Data from 5 studies were used to investigate different questions associating health status before breeding and reproductive responses. Health information for all studies was composed of the incidence of retained fetal membranes, metritis, mastitis, lameness, and respiratory and digestive problems from parturition until the day of breeding. Retained placenta and metritis were grouped as uterine disease (UTD). Mastitis, lameness, digestive and respiratory problems were grouped as nonuterine diseases (NUTD). Study 1 evaluated the effect of disease before artificial insemination (AI), anovulation before synchronization of the estrous cycle, and low body condition score at AI on pregnancy per AI, as well as their potential interactions or additive effects. Study 2 investigated the effect of site of inflammation (UTD vs. NUTD) and time of occurrence relative to preantral or antral stages of ovulatory follicle development, and the effect of UTD and NUTD on fertility responses of cows bred by AI or by embryo transfer. Study 3 evaluated the effect of disease on fertilization and embryonic development to the morula stage. Study 4 evaluated the effect of disease on preimplantation conceptus development as well as secretion of IFN-τ and transcriptome. Study 5 investigated the effect of diseases before AI on the transcript expression of interferon-stimulated genes in peripheral blood leukocytes during peri-implantation stages of conceptus development after first AI postpartum. Altogether, these studies demonstrated that inflammatory disease before breeding reduced fertilization of oocytes and development to morula, and impaired early conceptus development to elongation stages and secretion of IFN-τ in the uterine lumen. Diseases caused inflammation-like changes in transcriptome of conceptus cells, increased risk of pregnancy loss, and reduced pregnancy or calving per breeding. Moreover, the effects on reproduction were independent of cyclic status before synchronization of the estrous cycle and body condition score at breeding, which all had additive negative effects on fertility of dairy cows. Occurrence of disease at preantral or at antral stages of ovulatory follicle development had similar detrimental effects on pregnancy results. The carryover effects of diseases on developmental biology might last longer than 4 mo. Reduced oocyte competence is a likely reason for carryover effects of diseases on developmental biology, but impaired uterine environment was also shown to be involved.

The objectives of this study were to evaluate the effects of an intravaginal insert containing progesterone (CIDR, controlled internal drug releasing) administered in presynchronization and resynchronization protocols on cyclicity, detection of estrus, pregnancy rate, and pregnancy loss to first AI; reinsemination patterns; and pregnancy rates to second postpartum AI before and after the time of first-service pregnancy diagnosis in dairy cows. Holstein cows (n = 1,052) were blocked by parity and BCS at 3 +/- 3 d in milk (study d 0 = day of calving) and assigned randomly to 1 of 3 presynchronization treatments. During the presynchronization programs, all cows received 2 injections of PGF2alpha, on study d 35 and 49. Cows enrolled in the control presynchronization treatment received AI after detected estrus from study d 49 to 62. Cows enrolled in the CIDR estrus-detection (CED) presynchronization treatment received a CIDR insert from study d 42 to 49 and AI on detection of estrus from d 49 to 62. Cows enrolled in the CIDR timed AI (CTAI) presynchronization treatment received the same treatment as CED, but were subjected to timed AI on study d 72 after the Ovsynch (GnRH, 7 d PGF2alpha, 2 d GnRH, 24 h timed AI) protocol. The control and CED cows not inseminated by study d 62 were enrolled in the Ovsynch protocol on the same day and received timed AI on study d 72. After first AI, cows were assigned to no resynchronization (RCON) or resynchronization with a CIDR insert (RCIDR) between 14 and 21 d after AI. Blood samples collected on study d 35, 49, and 62 were analyzed for concentrations of progesterone and cows were classified as anestrous when progesterone was < 1.0 ng/mL in the first 2 samples. On study d 62, anestrous cows with progesterone > or = 1.0 ng/mL were classified as having resumed cyclicity. Pregnancy was diagnosed at 31 and 60 d after first AI and at 42 d after second AI. A greater proportion of anestrous cows in CED and CTAI became cyclic by d 62 postpartum than control cows. Resynchronization with the CIDR insert increased the pregnancy rate at 31 d after first AI in CED and CTAI, and at 60 d after AI in all cows because of reduced pregnancy loss. These results indicate that presynchronization with the CIDR insert increased induction of cyclicity in anestrous cows and that resynchronization with the CIDR insert did not affect the reinsemination rate but did reduce pregnancy loss and increased the pregnancy rate at 60 d after first AI.

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The objective of this study was to determine risk-factors affecting increase in physical activity during estrus and pregnancy per artificial insemination (P/AI) in lactating dairy cows. Cows were monitored continuously by 2 automated activity monitors [a collar-mounted accelerometer (HT; Heatime, SCR Engineers, Netanya, Israel) and a leg-mounted pedometer (BO; Boumatic Heat-seeker-TX, Boumatic Dairy Equipment, Madison, WI)]. When an increase in activity was detected, body condition score (BCS) and blood samples were collected, ovaries were scanned by ultrasonography, and, if the cow was eligible for breeding, artificial insemination was performed. Milk production and health-related data were recorded throughout the experimental period. Pregnancy diagnosis was performed at 42 ± 7 d of gestation. Data were analyzed using Pearson correlation, ANOVA, and logistic regression. A total of 1,099 true events of estrus from 318 lactating Holstein cows were recorded, averaging 3.46 ± 1.1 events per cow. Positive predictive value for estrus episodes detected by the HT and BO systems were 89.6 and 85.5%, respectively. Mean peak activity at estrus (PA) recorded by the HT system was 71.6 ± 20.7 index-value, and 334.3 ± 155.7% relative increase by the BO system. Compared with primiparous, multiparous cows expressed estrus with lower PA (69.3 ± 0.8 vs. 75.9 ± 1.1 index for HT; 323.9 ± 6.0 vs. 354.8 ± 8.48% for BO) and shorter duration (DU; 10.7 ± 0.2 vs. 12.0 ± 0.3 h); DU was measured by HT only. Lower BCS was associated with decreased PA measured by both systems, estrus DU, and P/AI. Peak activity was weakly correlated with milk production on the day of artificial insemination (r = -0.20); however, when categorized into quartiles, the highest-yield cows had lower PA and DU. Follicle diameter was not correlated with PA or DU, but cows with greater concentrations of estradiol had higher PA. Cows with greater PA in both systems had greater P/AI than those with lower PA (36.5 vs. 24.6% for HT, 33.5 vs. 21.4% for BO). In conclusion, measurements of estrus events captured by automated activity monitors are correlated with BCS, parity, and secondary behavior signs related to estrus. Surprisingly, estrus intensity and duration were only weakly correlated with milk production, preovulatory follicle diameter, and concentrations of estradiol at estrus. Cows that had measurements of high-intensity estrus were significantly more fertile than low-intensity estrus.

The objectives of this study were to determine the effects of incorporating a progesterone intravaginal insert (CIDR) between the day of GnRH and PGF2alpha treatments of a timed AI protocol using estradiol cypionate (ECP) to synchronize ovulation on display of estrus, ovulation rate, pregnancy rate, and late embryonic loss in lactating cows. Holstein cows, 227 from Site 1 and 458 from Site 2, were presynchronized with two injections of PGF2alpha on study d 0 and 14, and subjected to a timed AI protocol (100 mixrog of GnRH on study d 28, 25 mg of PGF2alpha on study d 35, 1 mg of ECP on study d 36, and timed AI on study d 38) with or without a CIDR insert. Blood was collected on study d 14 and 28 for progesterone measurements to determine cyclicity. Ovaries were scanned on d 35, 37, and 42, and pregnancy diagnosed on d 65 and 79, which corresponded to 27 and 41 d after AI. Cows receiving a CIDR had similar rates of detected estrus (77.2 vs. 73.8%), ovulation (85.6 vs. 86.6%), and pregnancy at 27 (35.8 vs. 38.8%) and 41 d (29.3 vs. 32.3%) after AI, and late embryonic loss between 27 and 41 d after AI (18.3 vs. 16.8%) compared with control cows. The CIDR eliminated cows in estrus before the last PGF2alpha injection and decreased (P < 0.001) the proportion of cows bearing a corpus luteum (CL) at the last PGF2alpha injection because of less ovulation in response to the GnRH and greater spontaneous CL regression. Cyclic cows had greater (P = 0.03) pregnancy rates than anovulatory cows at 41 d after AI (33.8 vs. 20.4%) because of decreased (P = 0.06) late embryonic loss (16.0 vs. 30.3%). The ovulatory follicle was larger (P < 0.001) in cows in estrus, and a greater proportion of cows with follicles > or = 15 mm displayed estrus (P< 0.001) and ovulated (P = 0.05) compared with cows with follicles <15 mm. Pregnancy rates were greater (P < 0.001) for cows displaying estrus, which were related to the greater (P < 0.001) ovulation rate and decreased (P = 0.08) late embryonic loss for cows in estrus at AI. Cows that were cyclic and responded to the presynchronization protocol (high progesterone at GnRH and CL at PGF2alpha) had the highest pregnancy rates. Incorporation of a CIDR insert into a presynchronized timed AI protocol using ECP to induce estrus and ovulation did not improve pregnancy rates in lactating dairy cows. Improvements in pregnancy rates in cows treated with ECP to induce ovulation in a timed AI protocol are expected when more cows display estrus, thereby increasing ovulation rate.

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