Comparison of reproductive management programs for submission of Holstein heifers for first insemination with conventional or sexed semen based on expression of estrus, pregnancy outcomes, and cost per pregnancy.

Our objective was to evaluate reproductive management programs for submission of Holstein heifers for first insemination with conventional or sexed semen. In experiment 1, nulliparous Holstein heifers (n = 462) were submitted to a 5-d progesterone-releasing intravaginal device (PRID)-Synch protocol [d 0, GnRH + PRID; d 5, PGF2α - PRID; d 6, PGF2α; d 8, GnRH + TAI] and were randomly assigned for PRID removal on d 5 or 6 of the protocol followed by timed artificial insemination (TAI) with conventional semen. Delaying PRID removal decreased early expression of estrus before scheduled TAI (0.9 vs. 12.2%), and pregnancies per AI (P/AI) did not differ between treatments. In experiment 2, nulliparous Holstein heifers (n = 736) from 3 commercial farms were randomized within farm to 1 of 3 treatments for first AI with sexed semen: (1) CIDR5 [d -6, GnRH + controlled internal drug release (CIDR); d -1, PGF2α - CIDR; d 0, PGF2α; d 2, GnRH + TAI]; (2) CIDR6 (d -6, GnRH + CIDR; d -1, PGF2α; d 0, PGF2α - CIDR; d 2, GnRH + TAI); and (3) EDAI (PGF2α on d 0 followed by once-daily estrous detection and AI). Delaying CIDR removal decreased early expression of estrus before scheduled TAI (0.004 vs. 27.8%); however, CIDR5 heifers tended to have more P/AI at 35 (53 vs. 45 vs. 46%) and 64 (52 vs. 45 vs. 45%) days after AI than CIDR6 and EDAI heifers, respectively. Overall, CIDR5 and CIDR6 heifers had fewer days to first AI and pregnancy than EDAI heifers which resulted in less feed costs than EDAI heifers due to fewer days on feed until pregnancy. Despite greater hormonal treatment costs for CIDR5 heifers, costs per pregnancy were $16.66 less for CIDR5 than for EDAI heifers. In conclusion, delaying PRID removal by 24 h within a 5-d PRID-Synch protocol in experiment 1 suppressed early expression of estrus before TAI, and P/AI for heifers inseminated with conventional semen did not differ between treatments. By contrast, although delaying CIDR removal by 24 h within a 5-CIDR-Synch protocol in experiment 2 suppressed early expression of estrus before TAI, delaying CIDR removal by 24 h tended to decrease P/AI for heifers inseminated with sexed semen. Further, submission of heifers to a 5-d CIDR-Synch protocol for first AI tended to increase P/AI and decrease the cost per pregnancy compared with EDAI heifers.

Patients' and rheumatologists' perspectives on the efficacy and safety of low-dose glucocorticoids in rheumatoid arthritis-an international survey within the GLORIA study.

OBJECTIVE: To evaluate the current perspectives of patients and health professionals regarding the efficacy and safety of low-dose glucocorticoids (GCs) in RA. METHODS: Two online surveys were disseminated to patients and health professionals, in their native language, through national patient organizations and national rheumatology medical societies, respectively. SurveyMonkey®, MediGuard.org and the Glucocorticoid Low-dose Outcome in RA Study (GLORIA) website were used to offer and deliver these surveys. RESULTS: A total of 1221 RA patients with exposure to GCs, and 414 rheumatologists completed the surveys. Patients and rheumatologists reported high levels of agreement regarding the efficacy of low-dose GCs: at least 70% considered that they are very rapid and effective in the control of signs and symptoms of RA. However, half of the patients also reported having suffered serious adverse events with GCs, and 83% described concerns about safety. The majority of rheumatologists estimated that endocrine, ophthalmologic and cutaneous adverse events affect >4% of all patients treated with low-dose GCs for 2 years, based on a heat map. CONCLUSIONS: RA patients with self-reported exposure to GCs express high levels of satisfaction with low-dose GCs efficacy, as do rheumatologists. However, both expressed excessive concerns regarding the safety of GCs (greatly exceeding the published evidence data), which may compromise the optimal use of this medication. This study indicates that there is an unmet need for appropriately designed prospective trials that shed light on the real risk associated with low-dose GCs, as well as a need for renovated educational programs on the real benefits and harms of low-dose GCs, for both patients and physicians.

Effect of manipulating progesterone before timed artificial insemination on reproductive and endocrine outcomes in high-producing multiparous Holstein cows.

Our objective was to evaluate the effect of manipulating progesterone (P4) concentrations before timed artificial insemination (TAI) on reproductive and endocrine outcomes in high-producing Holstein cows. Multiparous lactating Holstein cows (n = 80) were synchronized for first TAI using a Double-Ovsynch protocol and were randomly assigned to receive 25 mg of PGF2α 1 d after the first GnRH treatment of the Breeding-Ovsynch protocol that included a once-used P4 insert (low-P4 group) or to receive 2 new P4 inserts during the Breeding-Ovsynch protocol (high-P4 group). Blood samples were collected thrice weekly from -10 to 32 d relative to TAI for all cows and from 32 to 67 d after TAI for pregnant cows and were analyzed for P4 and pregnancy-specific protein B (PSPB) concentrations. Expression of IFNτ-stimulated gene 15 (ISG15) was assessed in blood leukocytes 18 and 20 d after TAI. As expected, P4 concentrations were greater for high-P4 cows than for low-P4 cows from 3 to 8 d before TAI. Incidence of double ovulation was 3-fold greater for low-P4 cows than for high-P4 cows (33 vs. 10%), which resulted in more twin pregnancies 32 d after TAI for low-P4 cows than for high-P4 cows (29 vs. 0%). Low-P4 cows had larger preovulatory follicles at the last GnRH treatment of the Double-Ovsynch protocol and greater P4 concentrations than high-P4 cows after TAI. Relative expression of ISG15 mRNA 18 and 20 d after TAI was greater for low-P4 cows than for high-P4 cows and for pregnant cows than for nonpregnant cows. Overall, PSPB concentrations tended to be greater for low-P4 cows than for high-P4 cows, and pregnant cows had greater P4 concentrations than nonpregnant cows. In summary, cows with low P4 before TAI had increased preovulatory follicle diameter, PSPB concentrations, relative expression of ISG15 mRNA 18 and 20 d after TAI, double ovulations, and twinning compared with cows with high P4 before TAI. Increasing P4 before TAI may effectively decrease double ovulation and twinning in high-producing multiparous Holstein cows.

Ovulation and fertility response to commercially available GnRH products in lactating cows synchronized with the Double-Ovsynch protocol.

This study was designed to evaluate whether commonly used gonadorelin products that are commercially available in the United States results in comparable ovulation and pregnancy per AI (P/AI) in synchronized lactating dairy cows. A total of 1411 Holstein cows receiving a Double-Ovsynch protocol (DOV) for conducting the first postpartum AI were randomized to receive one of the following GnRH products throughout the Double-Ovsynch: 1) Cystorelin® (CYS, gonadorelin diacetate tetrahydrate, n = 484); 2) Factrel® (FAC, gonadorelin hydrochloride, n = 482) or; 3) Fertagyl® (FER, gonadorelin diacetate tetrahydrate, n = 515). A subgroup of cows (n = 487) received ovarian ultrasound exams and collection of blood samples for progesterone (P4) analysis. Proportion of cows ovulating following the 3rd GnRH of DOV tended (P = 0.07) to differ between GnRH salts (hydrochloride = 61.5% vs. diacetate = 72.7%) but was similar for GnRH products (FER = 74.1% vs. FAC = 61.5% vs. CYS = 72.2%). Interestingly, a logistic regression analyses that considered the circulating P4 at the time of GnRH treatment indicated lower ovulation responses to FAC compared to FER and CYS; although greater circulating P4 decreased ovulation response to all GnRH products. Results for P/AI at 60 d post-insemination differed between GnRH salts (P = 0.02) as well as GnRH products (FER = 47.8% vs. FAC = 42.0% vs. CYS = 49.8%; P = 0.04). In conclusion, fertility following use of the Double-Ovsynch was less following a hydrochloride-based GnRH product likely due to lesser ovulatory responses throughout the synchronization protocol.

Effect of treatment with human chorionic gonadotropin 7 days after artificial insemination or at the time of embryo transfer on reproductive outcomes in nulliparous Holstein heifers.

Our objective was to assess the effect of treatment with human chorionic gonadotropin (hCG) 7 d after artificial insemination (AI) or at the time of in vitro-fertilized (IVF) embryo transfer on reproductive outcomes, including progesterone (P4), interferon-tau stimulated gene 15 (ISG15), pregnancy-specific protein B (PSPB), and pregnancies per AI (P/AI) or pregnancies per embryo transfer (P/ET), in nulliparous Holstein heifers. Heifers in experiment 1 were randomly assigned to receive no treatment (control; n = 129) or 2,000 IU of hCG 7 d after AI to a detected estrus (estrus = experimental d 0; hCG; n = 132). Heifers in experiment 2 were randomly assigned to receive no treatment (control; n = 143) or 2,000 IU of hCG (hCG; n = 148) at transfer of an IVF embryo 7 d after the last GnRH treatment of a 5-d controlled internal drug release-synch protocol (last GnRH = experimental d 0). Blood samples were collected from a subgroup of heifers (experiment 1, n = 82; experiment 2, n = 104) at d 7, 11, 18, 20, 25, 28, and 32, and blood samples from heifers diagnosed pregnant were collected on d 35, 39, 46, 53, 60, and 67. Blood samples were assayed for P4 by RIA and for PSPB by ELISA, and expression of ISG15 was assessed in mRNA isolated from blood leukocytes on d 18 and 20. Data were analyzed by ANOVA and logistic regression using the MIXED and GLIMMIX procedures. In both experiments, treatment with hCG increased P4 concentrations from d 11 to 32; however, treatment did not affect P/AI or P/ET at d 32 or 67, PSPB concentrations from d 11 to 67 of pregnancy, or relative ISG15 mRNA concentrations on d 18 or 20. Heifers diagnosed not pregnant at d 32 in experiment 2 with an extended luteal phase (>20 d) and treated with hCG had greater relative ISG15 mRNA concentrations on d 20 than control heifers. Treatment with hCG did not affect pregnancy loss in experiment 1, whereas heifers treated with hCG at the time of IVF embryo transfer had fewer pregnancy losses from d 32 to 67 than control heifers. We concluded that treatment with 2,000 IU of hCG 7 d after AI or at the time of embryo transfer increased P4 concentrations without affecting P/AI or P/ET in nulliparous Holstein heifers.

Short communication: Effect of adding a second prostaglandin F2α injection during the Ovsynch protocol on luteal regression and fertility in lactating dairy cows: A meta-analysis.

Incomplete luteal regression after treatment with a single dose of prostaglandin F2α during an Ovsynch protocol decreases fertility to timed artificial insemination (TAI). To increase the proportion of cows with complete luteal regression and subsequently pregnancy per artificial insemination (P/AI), an additional treatment with PGF2α 24 h after the first has been recommended. A systematic review of the literature and meta-analytical assessment were performed with the objective of evaluating the effects of adding a second PGF2α treatment during the Ovsynch protocol on luteal regression and reproductive performance in lactating dairy cows. Based on the heterogeneity among the experimental treatments, a fixed or a random effects meta-analysis was conducted. Reproductive outcomes of interest were luteal regression at the end of the Ovsynch protocol, and P/AI measured 32 to 39 d after TAI. Seven randomized controlled experiments from 6 published manuscripts including 5,356 cows with the primary objective to evaluate the effect of an additional treatment with PGF2α during the Ovsynch protocol on P/AI were used. Information regarding luteal regression at the end of the Ovsynch protocol was available for 1,856 cows. Adding a second PGF2α treatment on d 8 during the Ovsynch protocol increased the relative risk (RR) of complete luteal regression at the end of the Ovsynch protocol (RR = 1.14; 95% confidence interval = 1.10 to 1.17) using a fixed effects model and the RR for pregnancy (RR = 1.14; 95% confidence interval = 1.06 to 1.22) 32 d after TAI using a fixed effects model. No heterogeneity was observed among the 6 manuscripts regarding complete luteal regression and P/AI. In summary, there was a clear benefit of an additional PGF2α treatment during the Ovsynch protocol on luteal regression (+11.6 percentage units) and on P/AI (+4.6 percentage units).

Development of fertility programs to achieve high 21-day pregnancy rates in high-producing dairy cows.

Our objective is to overview the research that lead to the development of fertility programs for high-producing lactating dairy cows using only GnRH and Prostaglandin F2α (PGF2α), such as Ovsynch with a focus on the role of progesterone in fertility. A key factor affecting fertility to timed-AI is the response to each hormonal treatment of the Ovsynch protocol. Although not required for fertility, cows ovulating to the first GnRH treatment of the Ovsynch protocol (G1) have greater P/AI than cows failing to ovulate. The association between progesterone concentrations at each treatment during the Ovsynch protocol and fertility is reviewed, and data from 7792 cows from 14 studies are presented. Overall, medium P4 (between 0.5 and 6 ng/mL) at G1, high P4 concentrations (>1.0 ng/mL) at the PGF2α, and low P4 concentrations (<0.4 ng/mL) at final GnRH (G2) are associated with greater P/AI. The use of presynchronization treatments that increase the percentage of cows initiating the Ovsynch protocol during early diestrous increases the percentage of cows with medium P4 concentrations at G1 and with high P4 concentration at PGF2α. Some cows, however, fail to completely undergo luteal regression after a single PGF2α treatment, particularly cows that initiate the Ovsynch protocol in a low P4 environment and cows with a young (d 6) CL at the time of treatment with PGF2α. Addition of a second PGF2α treatment increased the percentage of cows with complete luteal regression and P/AI. The use of fertility programs that include the concepts described in this review have resulted in more P/AI than inseminating cows after an induced estrus at first insemination. In addition, extending these concepts to strategies for resynchronization of ovulation and the implementation of an aggressive reproductive management program for first and subsequent inseminations results in reproductive performance that is unprecedented for high-producing Holstein dairy cows.

Effect of dose and timing of prostaglandin F2α treatments during a Resynch protocol on luteal regression and fertility to timed artificial insemination in lactating Holstein cows.

Our objective was to evaluate the effect of a second PGF2α treatment (25 mg of dinoprost) or a double dose of PGF2α (50 mg of dinoprost) during a Resynch protocol on luteal regression and pregnancies per artificial insemination (P/AI) in lactating dairy cows. Lactating Holstein cows (n = 1,100) were randomly assigned at a nonpregnancy diagnosis to receive (1) Ovsynch (control: 100 µg of GnRH; 7 d, 25 mg of PGF2α; 56 h, 100 µg of GnRH), (2) Ovsynch with a second PGF2α treatment (GPPG: 100 µg of GnRH; 7 d, 25 mg of PGF2α; 24 h, 25 mg of PGF2α; 32 h, 100 µg of GnRH), or (3) Ovsynch with a double dose of PGF2α (GDDP: 100 µg of GnRH; 7 d, 50 mg of PGF2α; 56 h, 100 µg of GnRH). All cows received timed artificial insemination (TAI) approximately 16 h after the second GnRH treatment (G2). Pregnancy diagnosis was performed by transrectal palpation 39 ± 3 d after TAI, and pregnancy status was reconfirmed 66 d after TAI. Blood samples collected from a subset of cows in each treatment at the first PGF2α treatment (n = 394) and at G2 (n = 367) were assayed for progesterone (P4). Data were analyzed by logistic regression using the GLIMMIX procedure of SAS (SAS Institute Inc., Cary, NC). At 39 d after TAI, GPPG cows tended to have more P/AI than control cows [35% (137/387) vs. 31% (107/349)], whereas P/AI for GDDP cows [32% (118/364)] did not differ from that for control cows. Pregnancy loss from 38 to 66 d did not differ among treatments and was 8% (30/362). The percentage of cows with complete luteal regression (P4 <0.4 ng/mL at G2) tended to differ among treatments and was greater for GPPG cows than for GDDP and control cows (94% vs. 88% vs. 88%, respectively). Overall, cows with P4 <1 ng/mL at the first PGF2α treatment had fewer P/AI than cows with P4 ≥1 ng/mL (27% vs. 38%), whereas cows with P4 ≥0.4 ng/mL at G2 had fewer P/AI than cows with P4 <0.4 ng/mL (15% vs. 38%). We conclude that adding a second PGF2α treatment 24 h after the first within a Resynch protocol tended to increase the proportion of cows undergoing complete luteal regression and P/AI, whereas treatment with a double dose of PGF2α at a single time did not.

Characterization of luteal dynamics in lactating Holstein cows for 32 days after synchronization of ovulation and timed artificial insemination.

Approximately 20 to 30% of cows diagnosed not pregnant 32 d after timed artificial insemination (TAI) lack a corpus luteum (CL), and cows submitted to a resynchronization protocol in the absence of a CL have about 10% fewer pregnancies per AI (P/AI) than cows with a CL. An understanding of luteal dynamics after synchronization of ovulation and TAI may help refine strategies for reinseminating cows failing to conceive. Lactating Holstein cows (n = 141) were synchronized for first TAI using a Double-Ovsynch protocol. Thrice weekly from 4 to 32 d after TAI, blood samples were collected for evaluation of plasma progesterone (P4) concentrations, and CL diameter was measured using transrectal ultrasonography. Pregnancy status was determined using transrectal ultrasonography 32 d after TAI. Nonsynchronized cows (n = 4) were removed from the study. For cows diagnosed pregnant 32 d after TAI (n = 57), P4 increased from 4 to 15 d and then remained constant until 32 d after TAI, whereas CL volume increased from 4 to 11 d and then remained constant until 32 d after TAI. For cows diagnosed not pregnant 32 d after TAI (n = 80), P4 profiles were evaluated using statistical cluster analysis based on the day after TAI that P4 decreased to <1 ng/mL, resulting in 5 clusters: (1) CL regression 15 d after TAI (1.3%), (2) CL regression 18 to 22 d after TAI (55.0%), (3) CL regression 25 to 27 d after TAI (17.5%), (4) CL regression 29 to 32 d after TAI (5.0%), and (5) CL maintained until 32 d after TAI (21.3%). Plasma pregnancy-associated glycoprotein (PAG) levels at 25 and 32 d after TAI differed among clusters and were below the cut-off value of the assay for the classification of cows as not pregnant for cows in clusters 2, 3, and 4, whereas more than half of the cows in cluster 5 had increased plasma PAG levels. We conclude that at least half of the nonpregnant cows that maintained their CL until 32 d after TAI were initially pregnant but underwent early pregnancy loss based on increased plasma PAG levels at 25 and 32 d after TAI.

Fertility of lactating Holstein cows submitted to a Double-Ovsynch protocol and timed artificial insemination versus artificial insemination after synchronization of estrus at a similar day in milk range.

Our objective was to compare the AI submission rate and pregnancies per artificial insemination (P/AI) at first service of lactating Holstein cows submitted to a Double-Ovsynch protocol and timed artificial insemination (TAI) versus artificial insemination (AI) to a detected estrus after synchronization of estrus at a similar day in milk range. Lactating Holstein cows were randomly assigned to receive their first TAI after a Double-Ovsynch protocol (DO; n = 294) or to receive their first AI after a synchronized estrus (EST; n = 284). Pregnancy status was determined 33 ± 3 d after insemination and was reconfirmed 63 ± 3 d after insemination. Data were analyzed by ANOVA and logistic regression using the MIXED and GLIMMIX procedures of SAS (SAS Institute Inc., Cary, NC). By design, days in milk at first insemination did not differ between treatments (76.9 ± 0.2 vs. 76.7 ± 0.3 for DO vs. EST cows, respectively), but more DO cows were inseminated within 7 d after the end of the voluntary waiting period than EST cows (100.0 vs. 77.5%). Overall, DO cows had more P/AI than EST cows at both 33 d (49.0 vs. 38.6%) and 63 d (44.6 vs. 36.4%) after insemination, but pregnancy loss from 33 to 63 d after insemination did not differ between treatments. Primiparous cows had more P/AI than multiparous cows 33 and 63 d after insemination, but the treatment by parity interaction was not significant. Synchronization rate to the hormonal protocols was 85.3%, which did not differ between treatments; however, synchronized DO cows had more P/AI 33 d after insemination than synchronized EST cows (54.7 vs. 44.5%). In summary, submission of lactating Holstein cows to a Double-Ovsynch protocol and TAI for first insemination increased the percentage of cows inseminated within 7 d after the end of the voluntary waiting period and increased P/AI at 33 and 63 d after first insemination resulting in 64 and 58% more pregnant cows, respectively, than submission of cows for first AI after detection of estrus at a similar day in milk range. We conclude that, because the proportion of synchronized cows did not differ between treatments, DO cows had more P/AI than EST cows because of an intrinsic increase in fertility after submission to a fertility program.

Association of changes among body condition score during the transition period with NEFA and BHBA concentrations, milk production, fertility, and health of Holstein cows.

Our objective was to evaluate the association between body condition score (BCS) change during the transition period with fertility, non-esterified fatty acids (NEFA) and beta-hydroxybutyrate (BHBA) concentrations, milk yield, and health problems of Holstein cows in a retrospective cohort study. Holstein cows (n = 232) were assessed for BCS (5 point scale; 0.25 point increments) and had blood collected at 21 and 7 d before, on the day of, and 7 and 21 d after calving. Blood samples were assayed for NEFA and BHBA concentrations. All cows received a timed artificial insemination (TAI) at 65 ± 3 days in milk (DIM) following a Presynch-Ovsynch protocol with a progesterone implant during the Ovsynch protocol. Cows were grouped based on BCS change after calving as to whether they: 1) lost (L), 2) maintained (M), or 3) gained (G) BCS. Data were analyzed by logistic regression with GLIMMIX and ANOVA with repeated measures using the MIXED procedures of SAS. Both NEFA and BHBA concentrations after calving differed (P < 0.01) for cows that lost, maintained, or gained BCS from 21 d before to 21 d after calving (NEFA: 0.51 ± 0.01; 0.45 ± 0.01; 0.42 ± 0.01 mmol/L; BHBA: 0.73 ± 0.02; 0.70 ± 0.02; 0.68 ± 0.02 mmol/L; respectively; mean ± SEM). By design, BCS change after calving differed (P < 0.01) among groups and was -0.38; 0.00; and 0.35 for cows in groups L, M, and G, respectively. At 21 d before and 21 d after calving, BCS differed (P < 0.01) among groups and was [before (2.97, 2.70, and 2.57) and after (2.54, 2.70, and 2.90)] for cows in groups L, M, and G, respectively. Between evaluated days, higher circulating NEFA and BHBA concentrations were observed 7 d after calving. Change in BCS affected (P < 0.01) pregnancy/AI (P/AI), days to first ovulation, and percentage of cyclic cows at 50 DIM. At 32 d after TAI, P/AI differed (P < 0.01) for cows that lost [18% (11/84)], maintained [33% (26/80)], or gained [47% (32/68)] BCS. Cows that lost BCS during the transition period had more health events (P < 0.01), than cows that gained or maintained BCS. In conclusion, changes in BCS during the transition period affected NEFA and BHBA concentrations, fertility, and occurrence of health problems during the lactation.

Temporarily decreasing progesterone after timed artificial insemination decreased expression of interferon-tau stimulated gene 15 (ISG15) in blood leukocytes, serum pregnancy-specific protein B concentrations, and embryo size in lactating Holstein cows.

Our objective was to evaluate the effects of temporarily decreasing progesterone (P4) after timed artificial insemination (TAI) on embryonic growth in dairy cows. Lactating Holstein cows (n = 80) were submitted to a Double-Ovsynch protocol for first TAI and were assigned randomly to receive 12.5 mg of PGF2α 5 d after the last GnRH treatment (LowP4) or remain untreated (control). Blood samples were collected thrice weekly from 5 to 29 d after TAI for all cows and from 32 to 67 d for pregnant cows, and were analyzed for P4 and pregnancy-specific protein B concentrations. Expression of interferon-tau stimulated gene 15 (ISG15) was assessed in blood leukocyte mRNA 18 and 20 d after TAI. Pregnancy diagnosis was performed weekly using ultrasound from 32 to 67 d after TAI, and embryonic crown-rump length was measured 32, 39, and 46 d after TAI. Data were analyzed by ANOVA and logistic regression using the MIXED and GLIMMIX procedures of SAS. The LowP4 cows had less P4 than control cows from 6 to 11 d after TAI; however, pregnancy outcomes 32 d after TAI and pregnancy loss from 32 to 67 d after TAI did not differ between treatments. Control cows diagnosed pregnant 32 d after TAI had greater expression of ISG15 20 d after TAI than LowP4 cows diagnosed pregnant 32 d after TAI, and pregnant control cows had greater pregnancy-specific protein B concentrations from 25 to 67 d after TAI than pregnant LowP4 cows. Embryo size did not differ between treatments 32 and 39 d after TAI, but control cows had larger embryos 46 d after TAI. In conclusion, temporarily decreasing P4 after TAI decreased embryonic growth during early pregnancy in lactating Holstein cows but did not affect pregnancies per artificial insemination or pregnancy loss.

Effect of manipulating progesterone before timed artificial insemination on reproductive and endocrine parameters in seasonal-calving, pasture-based Holstein-Friesian cows.

Fertility to timed AI (TAI) is profoundly affected by progesterone (P4) levels during hormonal synchronization protocols. Holstein-Friesian dairy cows managed in a seasonal-calving, pasture-based production system were randomly assigned to 2 treatments to manipulate P4 before TAI during growth of the preovulatory follicle. Cows in the first treatment (High P4; n=30) were submitted to a Double-Ovsynch protocol {Pre-Ovsynch [GnRH; 7 d, PGF2α; 3 d, GnRH] followed 7 d later by Breeding-Ovsynch [GnRH (G1); 7 d PGF2α; 24 h, PGF2α; 32 h, GnRH (G2); 16 h, TAI]}. Cows in the second treatment (n=30; Low P4) received the same Double-Ovsynch protocol but with an additional PGF2α treatment 24 h after G1. Overall, synchronization rate did not differ between treatments and was 92% (55/60). Unexpectedly, 37% of Low P4 cows were detected in estrus ~24 h before scheduled TAI and were inseminated ~16 h before scheduled TAI. Overall, P4 did not differ between treatments at G1, whereas High P4 cows had greater P4 concentrations at PGF2α and G2 than Low P4 cows. High P4 cows had the smallest mean follicle diameter at G2, whereas Low P4 cows with no estrus before TAI had intermediate mean follicle diameter at G2, and Low P4 cows with estrus before TAI had the largest mean follicle diameter. Low P4 cows with estrus before TAI had larger corpora lutea 15 d after TAI than Low P4 cows without estrus before TAI or High P4 cows. In accordance with corpus luteum size on d 15, High P4 cows and Low P4 cows without estrus before TAI had lower P4 from 4 to 46 d after TAI than Low P4 cows with estrus before TAI. Relative mRNA levels of the interferon-stimulated genes ISG15, MX1, MX2, and OAS1 were greater for Low P4 than for High P4 cows, whereas relative mRNA levels of RTP4 were greater for High P4 than for Low P4 cows 18 d after TAI. Treatment did not affect plasma pregnancy-associated glycoprotein concentrations after TAI; however, pregnancy-associated glycoprotein concentrations were affected by pregnancy status and parity. Treatment did not affect pregnancy per artificial insemination at 29, 39, or 60 d after TAI, and no pregnancy losses were observed from 39 to 60 d after TAI. We concluded that (1) Low P4 cows were more likely to express estrus than High P4 cows; (2) the subpopulation of Low P4 cows that expressed estrus had larger preovulatory follicles and greater P4 concentrations after TAI; and (3) regardless of estrus before TAI, all Low P4 cows had greater mRNA expression for 5 of 6 interferon-stimulated genes than High P4 cows 18 d after TAI.

Effect of Comedication With Conventional Synthetic Disease-Modifying Antirheumatic Drugs on Retention of Tumor Necrosis Factor Inhibitors in Patients With Spondyloarthritis: A Prospective Cohort Study.

OBJECTIVE: To evaluate whether use of comedication with conventional synthetic disease-modifying antirheumatic drugs (csDMARDs) influences the retention of tumor necrosis factor inhibitors (TNFi) in patients with spondyloarthritis (SpA). METHODS: Patients with SpA from the Rheumatic Diseases Portuguese Register who started treatment with their first TNFi between 2001 and 2014 were included in this study. Cox regression analysis was used to estimate the effect of comedication with csDMARDs on TNFi retention in 2 types of models: a model in which baseline (time-fixed) variables were included, and a second model incorporating time-varying variables, including sociodemographic features, measures of disease activity, measures of physical function, and cotreatment with other drugs (nonsteroidal antiinflammatory drugs and oral steroids). To control for possible confounding by indication, the effect of csDMARD comedication on TNFi retention was also tested after adjustment for the treatment propensity score. RESULTS: In total, 954 patients were included in the study, of whom 289 (30.3%) discontinued treatment with their first TNFi after a median follow-up time of 2.5 years (range 0.08-13 years). Inefficacy was the most common reason for TNFi discontinuation (55.7% of patients). In the multivariable analyses, comedication with csDMARDs had no measurable effect on TNFi retention, neither in the baseline model (hazard ratio [HR] 0.83, 95% confidence interval [95% CI] 0.59-1.16) nor during follow-up in the model adjusted for time-varying covariates (HR 1.07, 95% CI 0.68-1.68). The effect of csDMARD comedication remained nonsignificant after propensity score adjustment. CONCLUSION: Comedication with csDMARDs does not prolong TNFi retention in patients with SpA in clinical practice, suggesting that there is no benefit conferred by the concomitant use of these drugs.

[Dermatomyositis-like syndrome in x-linked agammaglobulinemia]. / SÍNDROMA DERMATOMIOSITE-LIKE EM DOENTE COM AGAMAGLOBULINÉMIA LIGADA AO CROMOSSOMA X.

Primary immunodeficiencies (PIDs) encompass more than 250 different pathological conditions. X-linked agammaglobulinemia (XLA) has been occasionally associated with cutaneous and muscular manifestations resembling dermatomyositis, often termed dermatomyositis-like syndrome (DLS). This syndrome has been associated with cutaneous, muscular and central nervous system manifestations, accompanying a persistent infection by an Echovirus. According to sixteen previously reported cases, this syndrome has a poor prognosis. We report the case of a 27-years old male, with XLA and DLS, successfully treated with 6 cycles of human immunoglobulin and methotrexate. Clinical symptoms improved dramatically with a complete resolution of the musculoskeletal manifestations. Despite this clinical response, prognosis should remain reserved. The evolution of this syndrome remains unpredictable and therapeutic options are limited. To the best of our knowledge, there are only a few reports of similar cases which have survived so many months after the diagnosis.

Adding a second prostaglandin F2α treatment to but not reducing the duration of a PRID-Synch protocol increases fertility after resynchronization of ovulation in lactating Holstein cows.

Our objective was to evaluate the effect of a second PGF2α treatment and duration of an Ovsynch protocol that included a progesterone-releasing intravaginal device (PRID) on progesterone (P4) concentrations and pregnancies per artificial insemination (P/AI) after resynchronization of ovulation and timed artificial insemination (TAI). Lactating Holstein cows (n=821) were assigned randomly at a nonpregnancy diagnosis (d 0) to 3 resynchronization protocols: (1) GnRH, d 0; PGF2α, d 7; GnRH, d 9.5 (7D1PGF); (2) GnRH, d 0; PGF2α, d 7; PGF2α, d 8; GnRH, d 9.5); (7D2PGF); or (3) GnRH, d 2; PGF2α, d 7; PGF2α, d 8; GnRH, d 9.5 (5D2PGF). All cows received a PRID at the first GnRH treatment of the resynchronization protocol, which was removed at the first PGF2α treatment, and all cows received TAI approximately 16h after the second GnRH treatment. Blood samples were collected from a subgroup of cows at each treatment of the resynchronization protocols. At 32 d after TAI, cows receiving a second PGF2α treatment (7D2PGF + 5D2PGF cows) had more P/AI (42.6 vs. 35.7%) than cows receiving a single PGF2α treatment (7D1PGF cows). For cows treated with a second PGF2α treatment, decreasing the duration of the protocol did not increase P/AI (41.4 vs. 43.8% for 7D2PGF vs. 5D2PGF cows). At 60 d after TAI, P/AI did not differ between cows treated with the 1 PGF2α (7D1PGF cows) or 2 PGF2α (7D2PGF + 5D1PGF cows) treatments (32.5 vs. 37.9%, respectively). In addition, reducing the duration of the protocol did not increase P/AI at 60 d after TAI (37.8 vs. 38.5% for 7D2PGF vs. 5D2PGF cows). Pregnancy loss from 32 to 60 d after TAI was not affected by the number of PGF2α treatments (8.5 vs. 10.6%, for 7D1PGF vs. 7D2PGF + 5D2PGF cows) or the duration of the protocol (9.1 vs. 12.1%, for 7D2PGF vs. 5D2PGF cows). The percentage of cows with incomplete luteal regression at the second GnRH treatment tended to differ among treatments and was lowest for 7D2PGF cows, intermediate for 5D2PGF cows, and greatest for 7D1PGF cows (1.9 vs. 6.9 vs. 11.0%, respectively). In conclusion, addition of a second PGF2α treatment tended to decrease the percentage of cows with incomplete luteal regression and increased P/AI 32 d after AI, whereas decreasing the duration of the Ovsynch protocol did not increase P/AI.

Short communication: Field fertility in Holstein bulls: Can type of breeding strategy (artificial insemination following estrus versus timed artificial insemination) alter service sire fertility?

The aim of this study was to compare pregnancy per artificial insemination (P/AI) from service sires used on artificial insemination after estrus detection (EAI) or timed artificial insemination (TAI) breedings. Confirmed artificial insemination outcome records from 3 national data centers were merged and used as a data source. Criteria edits were herd's overall P/AI within 20 and 60%, a minimum of 30 breedings reported per herd-year, service sires that were used in at least 10 different herds with no more than 40% of the breedings performed in a single herd, breeding records from lactating Holstein cows receiving their first to fifth postpartum breedings occurring within 45 to 375 d in milk, and cows with 1 to 5 lactations producing a minimum of 6,804 kg. Initially 1,142,859 breeding records were available for analysis. After editing, a subset of the data (n=857,539) was used to classify breeding codes into either EAI or TAI based on weekly insemination profile in each individual herd. The procedure HPMIXED of SAS was used and took into account effects of state, farm, cow identification, breeding month, year, parity, days in milk at breeding, and service sire. This model was used independently for the 2 types osires f breeding codes (EAI vs. TAI), and service sire P/AI rankings within each breeding code were performed for sires with >700 breedings (94 sires) and for with >1,000 breedings (n=56 sires) following both EAI and TAI. Correlation for service sire fertility rankings following EAI and TAI was performed with the PROC CORR of SAS. Service sire P/AI rankings produced with EAI and TAI were 0.81 (for sires with >700 breedings) and 0.84 (for sires with >1,000 breedings). In addition, important changes occurred in service sire P/AI ranking to EAI and TAI for sires with less than 10,000 recorded artificial inseminations. In conclusion, the type of breeding strategy (EAI or TAI) was associated with some changes in service sire P/AI ranking, but ranking changes declined as number of breedings per service sire increased. Future randomized studies need to explore whether changes in P/AI ranking to EAI versus TAI are due to specific semen characteristics.

Manipulation of progesterone to increase ovulatory response to the first GnRH treatment of an Ovsynch protocol in lactating dairy cows receiving first timed artificial insemination.

Ovulation to the first GnRH (G1) treatment of the Ovsynch protocol improves synchronization rate and pregnancies per AI (P/AI). Elevated progesterone (P4) concentrations at G1 decrease the ovulatory response by decreasing the magnitude of the GnRH-induced LH surge. Therefore, our objective was to evaluate the effect of temporarily decreasing P4 concentrations before initiation of an Ovsynch protocol on ovulatory response to G1 and P/AI. Lactating Holstein cows (n=800) at 53±3 (herd A) or 51±3 (herd B) d in milk (DIM) were synchronized using a modified Double-Ovsynch protocol [pre-Ovsynch protocol (d 0, GnRH; d 7, PGF2α; d 10, GnRH) followed 7 d later by an Ovsynch-56 protocol (d 0, G1; d 7, PGF2α; d 8, PGF2α; d 9.5, GnRH)] to receive first timed artificial insemination (TAI; 80±3 DIM) 16h after the last GnRH treatment. Cows were randomly assigned to receive 12.5mg of PGF2α (a half-dose of dinoprost tromethamine) 2 d before G1 (low-P4) or serve as untreated controls (high-P4). Overall, high-P4 cows had greater P4 concentrations at G1 compared with low-P4 cows (3.0 vs. 1.3ng/mL, respectively). Ovulatory response to G1 was greater for low-P4 than high-P4 cows [81.1 vs. 60.3%, respectively]. Premature luteal regression during the second Ovsynch protocol did not differ between treatments [15.0% vs. 10.7%; for low-P4 vs. high-P4 cows, respectively]. Overall, P/AI did not differ between treatments 32 d after TAI [56.3 vs. 52.9%, for low-P4 vs. high-P4 cows, respectively] or 67 d after AI [50.5 vs. 47.6%, for low-P4 vs. high-P4 cows, respectively]. Pregnancy loss from 32 to 67 d after TAI did not differ between treatments [9.9 vs. 9.3%, for low-P4 vs. high-P4 cows, respectively]. Overall, cows that ovulated to G1 had more P/AI than cows that did not ovulate [58.2 vs. 41.8%, respectively]. The increase in P/AI for cows that ovulated to G1 (16.4%) combined with the observed increase in ovulation to G1 due to treatment (20.8%; low-P4 - high-P4) resulted in the expected numerical increase in P/AI of 3.4% in low-P4 versus high-P4 cows observed in this experiment. We conclude that administration of a half-dose of PGF2α 2 d before G1 during a Double-Ovsynch protocol decreased P4 at G1 and increased ovulatory response to G1.

Modifications to Ovsynch improve fertility during resynchronization: Evaluation of presynchronization with gonadotropin-releasing hormone 6 d before initiation of Ovsynch and addition of a second prostaglandin F2α treatment.

Lactating Holstein cows (n=897) were randomly assigned to a 2×2 factorial arrangement of treatments to compare the main effects of presynchronization with GnRH 6 d before beginning an Ovsynch protocol (±GnRH) and a second PGF2α treatment 24h after the first (1 vs. 2 PGF2α) on pregnancies per artificial insemination (P/AI). This resulted in the following 4 treatments: (1) an Ovsynch protocol (GPG, control); (2) presynchronization with GnRH followed by an Ovsynch protocol (GGPG); (3) an Ovsynch protocol with a second PGF2α treatment (GPPG); and (4) presynchronization with GnRH followed by an Ovsynch protocol with a second PGF2α treatment (GGPPG). All cows were submitted for first timed artificial insemination (TAI) using a Presynch Ovsynch protocol, and cows detected in estrus after the second PGF2α treatment of the Presynch portion of the protocol were inseminated and removed from the experiment. Nonpregnant cows were resynchronized using an Ovsynch protocol initiated 32±3 d after artificial insemination. Blood samples were collected at the first GnRH treatment (G1), at the PGF2α treatment (PGF), and at the last GnRH treatment (G2) of the Ovsynch protocol and were assayed for progesterone (P4) concentrations. Overall, P/AI tended to be greater for cows receiving a second PGF2α treatment compared with cows not receiving the second PGF2α treatment (40 and 37% for GGPPG and GPPG treatments, respectively, vs. 33 and 32% for GGPG and GPG treatments, respectively). Interestingly, treatment effects on P/AI were detected only for resynchronized cows receiving second and greater TAI, but not for cows receiving first TAI. Fewer cows presynchronized with GnRH had low (<0.5ng/mL) P4 at G1 than cows not presynchronized (13 vs. 25%), and P4 was greater at PGF2α for cows presynchronized with GnRH (4.4 vs. 4.0ng/mL). Cows receiving 2 PGF2α treatments had lower P4 at G2 than cows receiving 1 PGF2α treatment (0.15 vs. 0.35ng/mL). Differences in P4 at PGF2α were detected only for resynchronized cows and not for cows submitted for first TAI. We conclude that presynchronization with GnRH 6 d before beginning an Ovsynch protocol tended to increase P4 at the first GnRH treatment of an Ovsynch protocol, and that a second PGF2α treatment 24h after the first decreased P4 at TAI, thereby increasing P/AI in cows resynchronized for second and greater TAI.

Factors associated with pregnancy-associated glycoprotein (PAG) levels in plasma and milk of Holstein cows during early pregnancy and their effect on the accuracy of pregnancy diagnosis.

Lactating Holstein cows (n = 141) were synchronized to receive their first timed artificial insemination (TAI). Blood and milk samples were collected 25 and 32 d after TAI, and pregnancy status was determined 32 d after TAI using transrectal ultrasonography. Cows diagnosed pregnant with singletons (n = 48) continued the experiment in which blood and milk samples were collected and pregnancy status was assessed weekly using transrectal ultrasonography from 39 to 102 d after TAI. Plasma and milk samples were assayed for pregnancy-associated glycoprotein (PAG) levels using commercial ELISA kits. Compared to ultrasonography, accuracy was 92% for the plasma PAG ELISA test and 89% for the milk PAG ELISA test 32 d after TAI. Plasma and milk PAG levels for pregnant cows increased from 25 d to an early peak 32 d after TAI. Plasma and milk PAG levels then decreased from 32 d after TAI to a nadir from 53 to 60 d after TAI for the plasma PAG assay and from 46 to 67 d after TAI for the milk PAG assay followed by an increase from 74 to 102 d after TAI. Overall, plasma PAG levels were approximately 2-fold greater compared with milk PAG levels, and primiparous cows had greater PAG levels in plasma and milk compared with multiparous cows. The incidence of pregnancy loss from 32 to 102 d after TAI based on ultrasonography was 13% for cows diagnosed with singleton pregnancies, and plasma and milk PAG levels decreased to nonpregnant levels within 7 to 14 d after pregnancy loss. Both plasma and milk PAG levels were negatively correlated with milk production for both primiparous and multiparous cows. We conclude that stage of gestation, parity, pregnancy loss, and milk production were associated with plasma and milk PAG levels after TAI similarly. Based on plasma and milk PAG profiles, the optimal time to conduct a first pregnancy diagnosis is around 32 d after AI, coinciding with an early peak in PAG levels. Because of the occurrence of pregnancy loss, all pregnant cows should be retested 74 d after AI or later when plasma and milk PAG levels in pregnant cows have rebounded from their nadir.