RESUMO
Pregnancy loss (PL) in lactating dairy cows disrupts reproductive and productive efficiency. We evaluated the expression of interferon-stimulated genes (ISG) in blood leukocytes, vaginal and cervical epithelial cells, luteolysis-related genes, progesterone, and pregnancy-associated glycoprotein (PAG) profiles in lactating dairy cows (n = 86) to gain insight about PL. Expression of ISG on d17, d19, and d21 was greater in cows that maintained the pregnancy (P33) compared to nonpregnant with no PL (NP). Greater ISG differences between groups were observed in the cervix (96.7-fold) than vagina (31.0-fold), and least in blood leukocytes (5.6-fold). Based on individual profiles of ISG and PAG, PL was determined to occur either before (~13%) or after (~25%) d22. For cows with PL before d22, ISG expression was similar on d17 but by d21 was lower and OXTR was greater than P33 cows and similar to NP; timing of luteolysis was similar compared to NP cows suggesting embryonic failure to promote luteal maintenance and to attach to the endometrium (no increase in PAG). For cows with PL after d22, ISG expression was similar to P33 cows on d17, d19, and d21 and luteolysis, when it occurred, was later than NP cows; delayed increase in PAG suggested later or inadequate embryonic attachment. In conclusion, PL before d22 occurred due to embryonic demise/failure to signal for luteal maintenance, as reflected in reduced ISG expression by d21. Alternatively, embryos with PL between d22 and 33 adequately signaled for luteal maintenance (ISG) but had delayed/inadequate embryonic attachment and/or inappropriate luteolysis causing PL.
Assuntos
Aborto Espontâneo , Interferons , Gravidez , Feminino , Humanos , Bovinos , Animais , Lactação , Inseminação Artificial/veterinária , Progesterona , GlicoproteínasRESUMO
Our objective was to determine the effect of inducing an accessory corpus luteum (CL) with human chorionic gonadotropin (hCG; 3,300 IU) on d 7 (hCG7) or 2 accessory CL with hCG on d 7 and 13 (hCG7+13) of the estrous cycle in noninseminated lactating Holstein cows. Cows (n = 86) between 39 and 64 DIM were pretreated with an Ovsynch + CIDR protocol, and only synchronized cows were used (n = 64). The day of the last GnRH of Ovsynch was considered d 0 of the estrous cycle. Follicular and luteal dynamics of cows were evaluated daily during an entire estrous cycle by ovarian ultrasonography. Blood samples were collected daily to measure serum concentration of progesterone (P4). Cows were randomly assigned to CON (n = 22, no treatment), hCG7 (n = 20), or hCG7+13 (n = 22) treatments. Two cows from hCG7+13 failed to ovulate after hCG and were removed from the analyses post-hCG treatment. The first day of luteolysis was considered the day that P4 declined to more than 2 SD of the mean for the 4 consecutive P4 concentrations with the greatest mean in late diestrus for each individual cow. The P4 cut-off for complete luteolysis was <1.0 ng/mL. Mean P4 on d 7 (3.23 ± 0.16 ng/mL) did not differ among treatments. Cows treated with hCG had greater total luteal and original CL volume and serum P4 during diestrus than CON. Cows treated with hCG7+13 had greater serum P4 after d 13 of the cycle than hCG7. Cycles were classified as having atypical cycles if the dominant follicle or future dominant follicle at the time of luteolysis did not ovulate (delayed ovulation; CON, n = 2; hCG7, n = 4; hCG7+13, n = 3), had a short cycle (CON, n = 1), delayed (CON, n = 2) or incomplete luteolysis (CON, n = 1; hCG7, n = 4; hCG7+13, n = 5). The remainder of cycles with normal complete luteolysis followed by ovulation were considered to be typical. Based on blood perfusion, the CON cow with incomplete luteolysis had 2 original CL remaining functional after first onset of luteolysis. The rest of the cows with incomplete luteolysis (9/10) had one or more CL regressing and at least one remaining functional after first onset of luteolysis. No specific pattern for CL side (ipsilateral vs. contralateral to a CL with complete regression) was observed for nonregressed CL. Cows with incomplete luteolysis had a second onset of luteolysis to undergo complete functional luteolysis. The proportion of cows with typical cycle was 73% (16/22) for CON, 60% (12/20) for hCG7, and 55% (11/20) for hCG7+13. Cows with typical cycles treated with hCG (hCG7 and hCG7+13) had a later onset of luteolysis, prolonged time to undergo complete luteolysis, and greater proportion of cows with 3 follicular waves than CON, resulting in a longer interovulatory interval for hCG7 and hCG7+13 than CON. In summary, accessory CL induced by hCG during diestrus not only altered follicular and luteal dynamics but also deferred and prolonged the luteolytic process.
Assuntos
Lactação , Luteólise , Animais , Bovinos , Gonadotropina Coriônica/farmacologia , Corpo Lúteo , Dinoprosta/farmacologia , Ciclo Estral , Sincronização do Estro , Feminino , Hormônio Liberador de Gonadotropina/farmacologia , ProgesteronaRESUMO
Because progesterone (P4) is essential for pregnancy establishment and maintenance, we investigated the effect of increased concentrations of P4 on embryonic attachment and concentrations of pregnancy-associated glycoproteins (PAG). Additionally, we investigated the relationships among luteal regression, pregnancy loss, and PAG concentrations in cows undergoing pregnancy loss by d 33 of pregnancy. Lactating dairy cows were allocated into control (n = 40) and human chorionic gonadotropin (hCG; 3,300 IU on d 7 and 13 to promote greater circulating P4; GnRH = d 0; n = 46) groups. Progesterone was measured daily from d 7 to 33, and PAG was measured daily from d 17 to 33; both hormones were also measured on d 47 and 61. An increase in PAG >10% compared with d 17 was considered a marker for pregnancy. The gold standard for pregnancy diagnosis was ultrasound evaluation of embryonic heartbeat on d 33. Statistical analyses were done with PROC MIXED from SAS Institute Inc. Concentrations of P4 were greater from d 8 onward in the hCG group. Concentrations of PAG did not differ between groups from d 17 to 33, suggesting no effect of increased P4 on hastening embryonic attachment and placental development. Nevertheless, PAG was greater in the hCG group on d 47 and 61, suggesting greater placental area or PAG secretory capacity. Pregnancy loss between d 20 and 33 occurred in 24.6% of cows. About 50% of pregnancy loss was due to luteal regression and about 50% was due to conceptus failure; that is, a decrease in PAG in the absence of luteal regression. In conclusion, increased P4 does not hasten embryonic attachment or early placental development but it leads to increased PAG in the second half of the second month of gestation. Additionally, pregnancy loss seems to be initiated by either corpus luteum regression or conceptus failure.
RESUMO
Our objective was to determine the effects of a single treatment of human chorionic gonadotropin (hCG) or GnRH from d 5 to 7 of the estrous cycle on cycle length, expression of estrus and fertility in lactating dairy cows. Lactating Holstein cows (n = 354) located in Farm 1 and lactating Jersey cows located in Farm 2 (n = 210) detected in estrus by an Automated Activity Monitor (AAM) system from 27 to 50 days in milk (DIM) were randomly assigned to receive one of three treatments from d 5 to 7 of the estrous cycle: control (untreated; CON; Holstein, n = 111; Jersey, n = 66), GnRH (86 µg gonadorelin acetate im; Holstein, n = 116; Jersey, n = 75), or hCG (3,300 IU im; Holstein, n = 127; Jersey, n = 69). Ovaries were scanned with ultrasound in a random subgroup of cows (Holstein/Farm 1, n = 147; Jersey/Farm 2, n = 94) on the day of treatment and 3 or 4 d later to determine ovulation. Estrus was detected after treatment by an AAM, and peak activity and heat index were recorded. A random subgroup of cows observed in estrus after treatment received first AI from 51 to 80 DIM (Holstein, n = 208; Jersey, n = 138). Pregnancy diagnoses were performed by transrectal ultrasonography at 37 ± 3 d post-AI. Holstein and Jersey cows treated with GnRH and hCG had an increased (P < 0.05) ovulatory response compared with controls. Human chorionic gonadotropin decreased (74%; P = 0.05) and GnRH tended to reduce (75%; P = 0.07) the proportion of multiparous Holstein cows returning to estrus compared with CON (86%). Cows treated with hCG had a longer (P < 0.01) estrous cycle length (24.6 ± 0.3 d, Holstein; 23.0 ± 0.3 d, Jersey) compared with CON cows (22.7 ± 0.3 d, Holstein; 21.3 ± 0.3 d Jersey) and GnRH (22.9 ± 0.3 d, Holstein; 21.1 ± 0.3 d Jersey). The percentage of cows with high (≥80) peak activity and heat index did not differ (P > 0.50) between treatments, and milk production did not affect (P > 0.65) the duration of estrus. Pregnancy per AI (P/AI) was not affected by treatments in Holstein (P = 0.93; CON: 34.3%, GnRH: 35.4%, and hCG: 31.5%) and in multiparous Jersey cows (P = 0.35; CON: 34.3%, GnRH: 35.4%, and hCG: 31.5%), but hCG had greater (P = 0.03; 55%) P/AI than GnRH (30.0%) and a trend (P = 0.06) for greater P/AI than CON (33.3%) in primiparous Jersey cows. In summary, inducing the formation of an accessory corpus luteum from d 5 to 7 of the estrous cycle with hCG reduced expression of estrus in multiparous Holstein cows. Moreover, hCG increased estrous cycle length in Holstein and Jersey cows, and it did not affect first service P/AI at 37 ± 3 d post-AI in Holstein and multiparous Jersey lactating cows. However, hCG increased P/AI in primiparous Jersey cows. Future research with a larger number of cows is needed to confirm these intriguing fertility results.
Assuntos
Gonadotropina Coriônica , Sincronização do Estro , Hormônio Liberador de Gonadotropina , Animais , Bovinos , Gonadotropina Coriônica/administração & dosagem , Ciclo Estral , Estro , Feminino , Fertilidade , Hormônio Liberador de Gonadotropina/administração & dosagem , Inseminação Artificial/veterinária , Lactação , Fase Luteal , Gravidez , ProgesteronaRESUMO
The objective of this study was to test the effect of low circulating concentrations of progesterone (P4) on pre-ovulatory follicle development in heifers as part of an overarching objective to develop a model to understand this phenomenon in dairy cattle without the confounding factors of lactation. Holstein heifers between 12 and 13 mo of age were pre-synchronized to ensure all heifers were on d 6 of the estrous cycle at the start of the Ovsynch program. Only heifers with CL regression and ovulation to the following pre-treatment strategy were used in the study: 0.5 mg cloprostenol (PGF2α), 2 d later, 0.1 mg GnRH, 6 d later, GnRH (G1; 1st GnRH of Ovsynch). Heifers (n = 159) responding to pre-treatment were randomly assigned to 4 groups and completed the Ovsynch program: high P4 control (HPC), low P4 control (LPC; PGF2α 24 h after G1), PG2 (PGF2α 24 and 48 h after G1) and PG3 (PGF2α 24, 48, and 96 h after G1). Only heifers that had ovulation to G1 remained in the study. Blood samples were collected in all heifers on d 7 (n = 157) and in a subset of heifers on d 1, 2, 3, 4 (n = 82) after G1 to measure serum P4. Pre-ovulatory follicle size at G1 (13.0 ± 0.1 mm; P = 0.53) and mean serum P4 24 h after G1 (d 1; 3.62 ± 0.11 ng/mL; P = 0.46) did not differ among treatments. HPC heifers had greater (P < 0.001) mean serum P4 compared to LPC, PG2 and PG3 on d 2, 3, 4, and 7. On d 2, 3 and 4, mean serum P4 of LPC, PG2 and PG3 heifers did not differ (P > 0.10). On d 7, LPC heifers had greater (P < 0.001) serum P4 compared to PG2 and PG3 heifers. Mean ± SEM serum P4 on d 7 after G1 was 8.43 ± 0.39, 2.55 ± 0.36, 1.58 ± 0.20, and 1.21 ± 0.15 ng/mL for HPC, LPC, PG2, and PG3, respectively. Percentage of heifers with P4 < 0.50 ng/mL on d 7 was greater (P < 0.05) for LPC, PG2 and PG3 (27, 32 and 26%, respectively) compared to HPC (0%). A greater (P < 0.05) proportion of heifers ovulated before G2 in the LPC, PG2 and PG3 than in the HPC. For heifers that ovulated after G2, low serum concentrations of P4 in LPC, PG2 and PG3 induced double ovulations in 6/97 heifers after the final GnRH of Ovsynch compared to 0/33 in HPC. In summary, PGF2α treatments during early CL development reduced circulating P4 concentrations 7 d after G1 compared with both HPC and LPC. However, it did not effectively control CL and follicle function to be utilized as a model to test high vs. low serum P4 on fertility parameters in Holstein heifers.