Association among lipopolysaccharide, the transforming growth factor-beta superfamily, follicular growth, and transcription factors in spontaneous bovine ovarian cysts.

The aim of this study was to investigate some of the growth and transcriptional factors originating from oocytes and granulosa cells in follicular fluid and to identify the relationships between the basic blood metabolite-metabolic hormones and intrafollicular lipopolysaccharide (LPS) concentrations. Thirty cows included in the study were allocated into 2 groups comprising 15 cows with healthy preovulatory follicles (cyclic cows) and 15 cows with confirmed cystic follicles. The ovaries and uteri of all cows were assessed by transrectal ultrasonographic examination. Blood serum samples were collected at 15, 25, 35, 45, and 55 d after calving for analysis of nonesterified fatty acids, ß-hydroxybutyrate, insulin, glucose, IGF-I, ACTH, and cortisol. Ovaries and uteri were examined using transrectal ultrasound. Vaginal discharge was evaluated on the same days. Follicular fluid was also aspirated on days 35-55 from the healthy preovulatory follicles and cystic follicles using a transvaginal ovum pickup method. The densitometric levels of inhibin-α, growth and differentiation factor (GDF-9), bone morphogenetic protein (BMP-6), and GATA-4 and GATA-6 proteins were analyzed by the Western blotting technique; the concentrations of antimullerian hormone (AMH), IGF-I, estradiol-17 beta (E2), and progesterone (P4) were determined by ELISA; and the concentrations of LPS in the follicular fluid were measured by the Limulus amebocyte lysate test. The serum insulin, ACTH, and cortisol concentrations were higher in cystic cows than cyclic cows, but serum IGF-I concentrations were lower in cystic cows. The IGF-I concentrations of cystic follicular fluids were lower, whereas AMH levels were significantly greater than those of healthy preovulatory follicular fluids. The cystic follicles had significantly lower expression levels of GDF-9, BMP-6, GATA-4, and GATA-6; in contrast, inhibin-α expression and LPS concentrations were significantly higher than in healthy preovulatory follicles. The proportion of pathologic vaginal discharge within 25 d postpartum in cystic cows were higher than in the cyclic group. In conclusion, it is suggested that intrafollicular dysregulation of the transforming growth factor-ß superfamily, growth, and transcriptional factors is affected by high intrafollicular LPS concentrations and systemic metabolic changes and these disturbances may be responsible for the generation of ovarian cysts.

Reducing the duration between gonadotropin-releasing hormone (GnRH) and prostaglandin F2α treatment in the Ovsynch protocol to 6 days improved ovulation to second GnRH treatment, but inclined to reduce fertility.

The aim of this study was to test Ovsynch (OVS) versus modified OVS (decreasing the interval between first GnRH and PGF2α to 6d) protocols on pregnancy per artificial insemination (AI) and OVS outcomes in cyclic dairy cows. Cyclic cows (n=920) were assigned to 1 of 2 groups: the OVS7 group (n=459) received the OVS protocol [GnRH treatment, PGF2α treatment 7d later, a second GnRH (GnRH2) treatment 56 h later, and timed AI (TAI) 16 to 18 h after the GnRH2 treatment], and the OVS6 group (n=461) received a modified OVS protocol, in which the interval between the first GnRH and PGF2α was decreased to 6d (GnRH treatment, PGF2α treatment 6d later, GnRH2 treatment 56 h later, and TAI 16 to 18h after the GnRH2 treatment). The response to the first GnRH of OVS was similar between OVS7 (54.5%, 250/459) and OVS6 (54.2%, 250/461) groups. The ovulatory response to GnRH2 of OVS was higher in OVS6 (91.3%, 421/461) than OVS7 (84.5%, 388/459). The follicle size (mean ± standard error of the mean) at the time of TAI was smaller in OVS6 (15.23 ± 0.11 mm) than OVS7 (16.04 ± 0.11 mm). When all cows were evaluated, the pregnancy per AI at 31 d tended to be lower in OVS6 (38.0%, 175/461) than in OVS7 (43.8%, 201/459). Moreover, the pregnancy per AI at 31 d was lower in OVS6 (40.9%, 172/421) compared with OVS7 (50.3%, 195/388) in synchronized cows. In conclusion, although the modified OVS protocol decreased the follicle size at the time of AI and increased the ovulatory response to GnRH2 of OVS, it unexpectedly reduced fertility in cyclic lactating dairy cows.

Effects of presynchronization with gonadotropin-releasing hormone-prostaglandin F2α or progesterone before Ovsynch in noncyclic dairy cows.

The aim of this study was to evaluate the efficiency of presynchronization with GnRH and PGF(2α) or with progesterone on overall Ovsynch (OVS) outcomes in noncyclic dairy cows. Cows were scanned 7 d apart with ultrasonography to determine cyclicity. Noncyclic cows (n=281; no corpus luteum on ovaries at both examinations) were randomly divided into 3 groups. In the GP group (n=108), the cows received GnRH and PGF(2α) (PGF) administrations 7 d apart, and OVS was started 11 d after PGF (GnRH-7 d-PGF-11 d-OVS). In the P4 group (n=90), the cows were treated for 7d with an intravaginal progesterone (P4) implant (PRID), and then OVS was started 11 d after removal of the implant (7d PRID-11 d-OVS). The control group (CON, n=83) did not receive any presynchronization, and OVS was started at the same time as in the other groups (18 d-OVS). The percentage of cows that became cyclic at the beginning of OVS was lower in the CON group (38.6%; 32/83) than in the presynchronization groups (66.7%, 72/108 in GP; 71.1%, 64/90 in P4). The response to the first GnRH of OVS did not differ among groups (63.9%, 53/83 in CON; 67.6%, 73/108 in GP; 63.3%; 57/90 in P4), and synchronization rates were similar among the groups (74-82%). The cows that responded to presynchronization treatments (GP or P4) had higher pregnancy per artificial insemination (P/AI) than did nonresponding cows. Pregnancy per AI at 31 d did not differ between groups (30.1%, 25/83 in CON; 43.5%, 47/108 in GP; and 35.6%, 32/90 in P4). However, CON cows (24.1%, 20/83) had lower P/AI at 62 d than GP cows (41.7%, 45/108). Embryonic loss was higher in CON (20%, 5/25) compared with the P4 group (3%, 1/32). The administration of GnRH followed by PGF or exogenous progesterone (PRID) similarly increased the percentage of cows that became cyclic before Ovsynch in noncyclic cows, but fertility did not improve. However, the cows that responded to presynchronization had higher fertility rates than the nonresponding cows.

Effect of presynchronization strategy before Ovsynch on fertility at first service in lactating dairy cows.

The aim of this study was to evaluate the effect of presynchronization with or without the detection of estrus on first service pregnancy per artificial insemination (P/AI) and on Ovsynch outcome in lactating dairy cows. A total of 511 cows were divided randomly but unevenly into 3 treatment groups at 44 to 50 days in milk (DIM). Ovsynch was started at the same time (69 to 75 DIM) in all three groups. Cows in the Ovsynch group (CON, N = 126) received no presynchronization before Ovsynch, and all cows were bred by timed AI (TAI). Cows in the presynchronization with estrus detection (PED) and the presynchronization with only TAI (PTAI) groups received two doses of prostaglandin F(2α) (PGF) 14 days apart, starting at 44 to 50 DIM. Ovsynch was initiated 11 days after the second PGF treatment. Cows in the PED group (N = 267) received AI if estrus was detected after either PGF injection. Cows that were not determined to be in estrus after PGF injection received Ovsynch and TAI. Cows in the PTAI group (N = 118) were not inseminated to estrus, with all cows receiving TAI after Ovsynch. The ovulatory response to the first GnRH injection administered as part of Ovsynch differed (P = 0.002) among treatment groups (83.1% in PTAI, 72.6% in PED, and 62.7% in CON). However, the ovulatory response to the second injection of GnRH during Ovsynch did not differ among treatment groups. Of the 267 PED cows, a total of 132 (49.4%) exhibited estrus and were inseminated. The P/AI at the 31-day pregnancy diagnosis was similar between the cows in the PED group with AI after estrus detection (37.9%; 50/132) and those bred with TAI (34.1%; 46/135). The P/AI in the CON group (46.8%; 59/126) was greater (P < 0.05) than that in the PED group (36.0%; 96/267). In addition, the P/AI in the CON group was greater (P = 0.04) than that in the PED cows receiving TAI (34.1%; 46/135) but less than that in the PED cows bred to estrus (37.9%; 50/132) (P = 0.16). At the 31-day pregnancy diagnosis, the cows in the PTAI group had greater P/AI (55.9%; 66/118) than both those in the PED group (P < 0.01; either estrus or TAI) and those in the CON group (P = 0.08). Thus, presynchronization with PGF (PTAI) increased the ovulatory response to Ovsynch and improved P/AI in dairy cows. Interestingly, the breeding of cows to estrus during presynchronization reduced fertility to the TAI and overall fertility, including cows bred to estrus and TAI. These results indicate that maximal fertility is obtained when all cows receive TAI after the presynchronization protocol.

Managing the dominant follicle in lactating dairy cows.

Reproductive efficiency is not optimal in high-producing dairy cows. Although many aspects of ovarian follicular growth in cows are similar to those observed in heifers, there are numerous specific differences in follicular development that may be linked with changes in reproductive physiology in high-producing lactating dairy cows. These include: 1) reduced circulating estradiol (E2) concentrations near estrus, 2) ovulation of follicles that are larger than the optimal size, 3) increased double ovulation and twinning, and 4) increased incidence of anovulation with a distinctive pattern of follicle growth in anovular dairy cows. The first three changes become more dramatic as milk production increases, although anovulation has not generally been associated with level of milk production. To overcome reproductive inefficiencies in dairy cows, reproductive management programs have been developed to synchronize ovulation and enable the use of timed AI in lactating dairy cows. Effective regulation of the CL, follicles, and hormonal environment during each part of the protocol is critical for optimizing these programs. This review discusses the distinct aspects of follicular development in lactating dairy cows and the methodologies that have been utilized in the past two decades in order to manage the dominant follicle during synchronization of ovulation and timed AI programs.

Ultrasonographic evaluation of endometrial thickness near timed AI as a predictor of fertility in high-producing dairy cows.

The objectives were to evaluate changes in endometrial thickness (ET) near the time of a synchronized ovulation and to assess the relationship of ET and fertility in lactating Holstein cows, with or without estrogen supplementation near timed ovulation. In Experiment 1, eight cows were examined with transrectal ultrasonography, once daily for 5 d, starting concurrent with PGF(2α) (PGF) treatment during an Ovsynch protocol (GnRH - 7d - PGF - 72h - GnRH). The ET increased rapidly after PGF (from ∼7 to ∼9.5 mm), remained > 9 mm for the next 2 d, then decreased to ∼8 and 7.4 mm, 1 and 2 d, respectively, after the second GnRH. In Experiment 2,642 cows (total of 758 breedings) were subjected to an Ovsynch protocol (GnRH - 7d - PGF - 56h - GnRH - 16h - timed AI); cows received either no further treatment (Ovsynch) or 1 mg of estradiol-17ß im 8 h before the second GnRH (Ovsynch + E2). For both uterine horns, ET was measured (∼2 cm from the internal uterine body bifurcation) before E2 treatment (48 h after PGF). In cows with ET ≤ 8 mm vs > 8 mm, rates of ovulation were 86.0% (n = 136) vs 98.1% (n = 472; P < 0.01), respectively, and percentage pregnant per AI (P/AI) were 26.7% (n = 146) vs 42.7% (n = 524; P < 0.01). Treatment with E2 increased P/AI in cows with lower ET (Ovsynch + E2 = 37.0% vs Ovsynch = 23.3%; P = 0.07), but did not significantly improve P/AI in cows with ET > 8 mm (Ovsynch + E2 = 43.4% vs Ovsynch = 42.1%). In conclusion, a single ultrasonographic evaluation of ET in Holstein cows 48 h after PGF treatment in an Ovsynch program was a good predictor of ovulation failure and pregnancy success. Perhaps poor fertility in cows with reduced ET was low peripheral E2 concentrations near AI, poor P4 priming, or luteolysis failure during timed AI procedures.

Effect of hCG vs. GnRH at the beginning of the Ovsynch on first ovulation and conception rates in cyclic lactating dairy cows.

Ovulatory response to the first GnRH of Ovsynch is a very important factor for determining the outcome of a successful synchronization. The aim of the present study was to develop a protocol to increase the percentage of cows that ovulated in response to the first administration of Ovsynch. This study was designed to compare ovulation rates in response to GnRH or hCG at the beginning of Ovsynch and to evaluate the effects of this manipulation on pregnancy. Cows (n = 371) with corpus luteum (CL) and at least one follicle greater than 10 mm diameter size on either ovary were included in the study. Cows were divided into two groups. The Ovsynch protocol began with GnRH (10 mug) in the GPG group (n = 161; GnRH-7d-PGF2alpha-56h-GnRH-18h-AI), whereas in the HPG group, the first GnRH of the Ovsynch was replaced with 1500 IU hCG (n = 210; hCG-7d-PGF2alpha-56h-GnRH-18h-AI). Ovarian ultrasonography was performed at the times of GnRH or hCG and of PGF2alpha administration, at the time of artificial insemination (AI) and seven days after AI, to determine ovulation. Maximal follicle size at the beginning of the Ovsynch did not affect on response to the first GnRH/hCG treatment. Conception rate (31 d) was 0.6 times more likely to be higher (P < 0.001) in cows that responded to the first hormonal administration of Ovsynch than in those that did not respond (95% CI = 0.29-0.71). Conception rate was found to be different between the HPG (37.6%, 79/210) and the GPG groups (48.4%, 78/161). Thus, beginning of the Ovsynch protocol with hCG did not increase ovulation and conception rate in lactating dairy cows, suggesting that hCG is not a suitable replacement of the first GnRH of Ovsynch. However, our results do show that increasing the ovulation rate in response to the first hormonal administration of Ovsynch can have a significant effect on conception rate.

Comparison of gonadorelin products in lactating dairy cows: efficacy based on induction of ovulation of an accessory follicle and circulating luteinizing hormone profiles.

This study evaluated whether the four gonadorelin products that are commercially available in the United States produce comparable ovulation responses in lactating cows. Dairy cows at 7 d after last gonadotropin-releasing hormone (GnRH) treatment of Ovsynch (Day 7), with a corpus luteum (CL) > or =15 mm and at least one follicle > or =10mm, were evaluated for response to GnRH treatment. Selected cows were randomized to receive (100 microg; im): (1) Cystorelin (n=146); (2) Factrel (n=132); (3) Fertagyl (n=140); or (4) Ovacyst (n=140). On Day 14, cows were examined for ovulation by detection of an accessory CL. Circulating luteinizing hormone (LH) concentrations were also evaluated in some cows after treatment with 100 microg (n=10 per group) or 50 microg (n=5 per group) GnRH. Statistical analyses were performed with the procedures MIXED and GLIMMIX of the SAS program. Percentage of cows ovulating differed (P<0.01) among groups, with that for Factrel being lower (55.3%) than that for Cystorelin (76.7%), Fertagyl (73.6%), or Ovacyst (85.0%). There was no effect of batch, parity, or follicle size on ovulation response, but increasing body condition score decreased ovulation response. There was a much greater LH release in cows treated with 100 microg than in those treated with 50 microg, but there were no detectable differences among products in time to LH peak, peak LH concentration, or area under the LH curve and no treatment effects nor treatment by time interactions on circulating LH profile. Thus, ovulation response to Factrel on Day 7 of the cycle was lower than that for other commercial GnRH products, although a definitive mechanism for this difference between products was not demonstrated.

Supplementation with estradiol-17beta before the last gonadotropin-releasing hormone injection of the Ovsynch protocol in lactating dairy cows.

The aim of this study was to determine whether an increase in circulating estrogen concentrations would increase percentage pregnant per artificial insemination (PP/AI) in a timed AI protocol in high-producing lactating dairy cows. We analyzed only cows having a synchronized ovulation to the last GnRH of the Ovsynch protocol (867/1,084). The control group (n = 420) received Ovsynch (GnRH--7 d--PGF(2alpha)--56 h--GnRH--16 h--timed AI). The treatment group (n = 447) had the same timed AI protocol with the addition of 1 mg of estradiol-17beta (E2) at 8 h before the second GnRH injection. Ovarian ultrasound and blood samples were taken just before E2 treatment of both groups. In a subset of cows (n = 563), pressure-activated estrus detection devices were used to assess expression of estrus at 48 to 72 h after PGF(2alpha) treatment. Ovulation was confirmed by ultrasound 7 d after timed AI. Treatment with E2 increased expression of estrus but overall PP/AI did not differ between E2 and control cows. There was an interaction between treatment and expression of estrus such that PP/AI was greater in E2-treated cows that showed estrus than in E2-treated or control cows that did not show estrus and tended to be greater than control cows that showed estrus. There was evidence for a treatment by ovulatory follicle size interaction on PP/AI. Supplementation with E2 improved PP/AI in cows ovulating medium (15 to 19 mm) but not smaller or larger follicles. The E2 treatment also tended to improve PP/AI in primiparous cows with low (< or =2.5) body condition score, and in cows at first postpartum service compared with Ovsynch alone. In conclusion, any improvements in PP/AI because of E2 treatment during a timed AI protocol appear to depend on expression of estrus, parity, body condition score, and size of ovulatory follicle.

Comparison of artificial insemination versus embryo transfer in lactating dairy cows.

Conception rates (CR) are low in dairy cows and previous research suggests that this could be due to impaired early embryonic development. Therefore, we hypothesized that CR could be improved by embryo transfer (ET) compared with AI. During 365 days, 550 potential breedings were used from 243 lactating Holstein cows (average milk production, 35 kg/day). Cows had their ovulation synchronized (GnRH-7d-PGF(2alpha)-3d-GnRH) and they were randomly assigned for AI immediately after the second GnRH injection (Day 0) or for transfer of one embryo 7 days later. Circulating progesterone concentrations and follicular and luteal size were determined on Days 0 and 7. Pregnancy diagnosis was performed on Days 25 or 32 and pregnant cows were reevaluated on Days 60-66. Single-ovulating cows with synchronized ovarian status had similar CR on Days 25-32 with ET (n = 176; 40.3%) and AI (n = 160; 35.6%). Pregnancy loss between Days 25-32 and 60-66 also did not differ (P = 0.38) between ET (26.2%) and AI (18.6%). When single (n = 334) and multiple (n = 57) ovulators were compared, independent of treatment, multiple ovulators had greater (P < 0.001) circulating progesterone concentrations on Day 7 (2.7 ng/ml versus 1.9 ng/ml) and there was a tendency (P = 0.10) for a greater CR in multiple ovulators (50.9% versus 38.1%). However, there was no difference in CR between AI and ET cows with multiple ovulations (50.0% versus 51.7%). In single-ovulating cows, CR tended to be lower for AI than ET in cows ovulating smaller follicles (diameter < or = 15 mm; 23.7% versus 42.3%; P = 0.06) but not average-diameter follicles (16-19 mm; 41.2% versus 37.3%; P = 0.81) or larger (> or =20 mm; 34.3 versus 51.0%; P = 0.36) follicles. Thus, although ET did not improve overall CR in lactating cows, follicle diameter and number of ovulating follicles may determine success with these procedures.

Production of lymphocyte-activating factors by mouse macrophages during aging and under the effect of short peptides.

Age-specific characteristics of production of lymphocyte-activating factor by mouse peritoneal macrophages and modulation of this production by short synthetic peptides (Vilon, Epithalon, and Cortagen) were studied. The production of lymphocyte-activating factors by macrophages stimulated with lipopolysaccharides in vitro was lower in old animals. The opposite modulating effects of short peptides on the production of lymphocyte-activating factors by resident and lipopolysaccharide-stimulated macrophages in young and old mice were demonstrated for the first time. This is a possible mechanism of immune system dysfunction during aging, which opens new vistas for its correction with short synthetic peptides.

Reduced dry periods and varying prepartum diets alter postpartum ovulation and reproductive measures.

There has been substantial recent interest in shortening dry periods; however, the effects of this management change on reproduction have not been adequately evaluated. Holstein cows (n = 58) were assigned in a randomized block design to 1 of 3 treatments: 1) traditional (T) dry period (approximately 56 d) in which cows were fed a low energy diet from 56 to 29 d prepartum followed by a moderate energy diet for 28 d; 2) shortened (S) dry period (approximately 28 d) in which cows were fed continuously a high energy diet; or 3) no planned (N) dry period in which cows were fed continuously a high energy diet. All cows received a high energy lactation diet after calving. Ovaries were evaluated by ultrasound and blood samples collected 3 times weekly beginning at d 6 or 7 postpartum until 7 d after second ovulation. Average days from calving until first detection of a 10-mm follicle were fewer in N (8.0 d) and S (8.9 d) than in T (10.5 d) cows. Time from calving to first ovulation was earlier in N (13.2 d) than in S (23.8 d) and T (31.9 d) cows. A greater percentage of follicles of the first follicular wave ovulated in N (89%; 16/18) than in T (42%; 8/19), with S (62%; 13/21) cows being intermediate. Double ovulation rate at the first ovulation was greater in T (61%) than N (16%), with S (35%) intermediate. No difference was detected in double ovulation rate at second ovulation (13/56). Number of cows with persistent corpus luteum (>30 d; 15/56) was not different among groups; however, short luteal phases were greater in N (28%; 5/18) than S (0%; 0/20) cows. Days to first artificial insemination were fewer in N (69.4 d) and S (68.0 d) than in T (75.0 d). First-service conception rate was greater in N (55%; 11/20) than in T (20%; 4/20), with S (26%; 6/23) cows being intermediate. Days open in pregnant cows were fewer in N (93.8 d) than in T (145.4 d), with S (121.2 d) cows being intermediate. Thus, shortening or eliminating the dry period leads to earlier postpartum ovulation and the results highlight the need for future large field studies to accurately evaluate the effect of dry period length on reproductive performance of lactating dairy cows.

Follicular size and response to Ovsynch versus detection of estrus in anovular and ovular lactating dairy cows.

In a commercial dairy herd, 316 lactating Holsteins were studied to determine the percentage of anovular cows, to examine follicular sizes in anovular cows, and to compare synchronized ovulation (Ovsynch) versus detection of estrus on fertility of ovular and anovular cows. Ultrasonography examinations at 47 to 53 d and at 54 to 60 d postpartum were used to measure follicles and to classify cows as ovular or anovular. Anovular cows were identified as those with no detectable luteal tissue by ultrasonography and by low progesterone in blood samples collected weekly. Anovular cows included 28% of 122 primiparous cows and 15% of 194 multiparous cows. Of 64 anovular cows, 20% had follicles > or = 25 mm that might be considered cystic (4% of total cows), 58% had 15- to 24-mm follicles, and 22% had 9- to 14-mm follicles. Cows identified as ovular and anovular were randomly assigned within cyclic status to one of two artificial insemination (AI) strategies: 1) AI after detected estrus during 21 d, or 2) timed AI after a 10-d Ovsynch protocol. Weekly ultrasonography continued for 21 d to detect ovulations. For the Ovsynch sub-groups, 97% of ovular and 94% of anovular cows ovulated after the second GnRH injection. Within 21 d, spontaneous ovulations for the detection of estrus sub-groups were 42% of anovular cows vs. 89% of ovular cows. Conception rates were greater for ovular cows regardless of treatment, but conception rates between respective Ovsynch and estrus detection groups for ovular (32%, 35%) or anovular (9%, 11%) cows were similar. Although 20% of lactating cows were not cyclic by about 60 d postpartum, nearly all ovulated following Ovsynch. However, anovular cows had lower conception than ovular cows whether inseminated after detected estrous or after Ovsynch.

A GnRH/LH surge without subsequent progesterone exposure can induce development of follicular cysts.

Our hypothesis was that follicular cysts would develop if cows experienced an estradiol-induced GnRH LH surge in the absence of an ovulatory follicle. Further, we hypothesized that estradiol would fail to induce a subsequent GnRH/LH surge in these cows until they were treated with progesterone. In experiment 1, seven cows were synchronized with a controlled internal drug releasing device (CIDR) for 9 d and each received 500 microg of cloprostenol on d 7. All follicles (> or = 5 mm in diameter) were aspirated at the time of CIDR removal using transvaginal follicular aspiration. Two days after aspiration, cows were treated with 5 mg of estradiol benzoate (EB) to induce a GnRH/LH surge in the absence of an ovulatory-sized follicle. All cows had an LH surge following the estradiol treatment and three of seven developed an anovulatory condition that resembled follicular cysts. The four cows that did not develop follicular cysts luteinized remaining cells from one aspirated follicle each. Thus, all cows with a progesterone elevation after the estradiol/GnRH/LH surge had subsequent ovulatory cycles, whereas the absence of progesterone was followed by follicular cysts. After 49 d, the anovulatory cows were induced back to normal cyclicity by insertion of a CIDR for 7 d. In two subsequent experiments, nine of 26 cows were induced to have follicular cysts by follicular aspiration followed by 5 mg of EB. After 26 d of observation, all cystic cows received a second treatment with 5 mg of EB and none of the cows showed an LH surge or ovulation. Cystic cows were untreated (n = 4 controls) or treated for 7 d with a CIDR (n = 5). All cystic cows were subsequently treated for a third time with 5 mg of EB. All CIDR-treated cows had an LH surge and ovulated, whereas none of the control cows had an LH surge or ovulation after the estradiol treatment. Thus, a large follicle anovulatory condition, similar to follicular cysts, can be induced by estradiol induction of a GnRH/LH surge in the absence of subsequent luteinization, and this condition prevents a GnRH/LH surge in response to high doses of estradiol. Progesterone eliminates this condition by reinitiation of GnRH/LH surges in response to estradiol.

Physiological classification of anovulatory conditions in cattle.

Evaluation of follicular growth patterns by ultrasound combined with measurement of circulating reproductive hormones has allowed designation of three functionally critical follicular sizes during the final stages of follicular growth: emergence (-4 mm), deviation (-9 mm), and ovulation (variable from 10 to 20 mm). Classification of anovulatory conditions on the basis of these three critical points is logical and provides for rational diagnosis and treatment of the underlying physiological condition. In extreme undernutrition, there is growth of follicles to emergence but not to deviation; however, the underlying pathophysiology is not defined because of relatively few scientific investigations of this condition. Anovulatory conditions with growth of follicles to deviation but not to ovulatory size have been extensively studied. Undernutrition and/or suckling can cause this anovulatory condition. It is characterized by a greater negative feedback effect of estradiol on GnRH/LH pulses than found in normally cycling cows. Another anovulatory condition that is common in high producing lactaing dairy cows is characterized by growth of follicles to larger than ovulatory size, such as is observed in cows with follicular cysts. This condition is characterized by an insensitivity of the hypothalamus to the positive feedback effects of estradiol. Thus, these last two common anovulatory conditions appear to be primarily due to changes in the responsiveness of the hypothalamus to estradiol. Treatments that increase circulating progesterone concentrations can help in the treatment of these two conditions by potentially altering GnRH/LH pulses and allowing the final stages of follicular growth or resetting the hypothalamic responsiveness to the positive feedback effects of estradiol.

Extending the postpartum anovulatory period in dairy cattle with estradiol cypionate.

Estradiol cypionate (ECP), a long-acting estrogen, has been used therapeutically in early postpartum (PP) dairy cows. In experiment 1, effects of ECP on circulating reproductive hormones, cyclicity, and ovarian function in early PP dairy cows were investigated. Lactating Holsteins received 10 mg of ECP (ECP; n = 17) or placebo (CON; n = 16) on d 7 PP. Serum and ultrasound data were acquired from 5 to 90 d of lactation. Compared to CON, ECP cows had greater serum estradiol for 10 d and lower serum FSH for 15 d posttreatment. After ECP, the appearance of follicles > or = 10 mm and time to first ovulation were delayed. Nevertheless, by 90 d PP, normal estrous cycles were found in only 50% of CON versus 88% of ECP cows. Primiparous, but not multiparous, cows receiving ECP had higher milk yields. Experiment 2 investigated effects of ECP on reproduction and milk production on a commercial dairy. Cows were blocked by parity and randomized to three treatments: 0 (n = 85), 4 (n = 85), or 10 (n = 86) mg of ECP on d 5 to 8 PP. Data included cycling status (two ultrasound examinations at 30 to 33 d PP and 7 d later), individual reproduction records, and daily milk yields from 10 to 90 d PP. In primiparous cows, ECP had no significant effects on ovulatory status or milk yields. By 40 d PP, a greater percentage of multiparous cows receiving 10 mg of ECP remained anovulatory compared with those receiving 0 or 4 mg. Milk yields were highest for multiparous cows receiving 4 mg of ECP, intermediate for the 10-mg dose, and lowest for controls. Lower conception was observed in multiparous cows receiving 4 mg of ECP. In summary, ECP delayed time to first ovulation particularly in multiparous cows, an effect associated with observed inhibition of circulating FSH. Milk yield responses to ECP were inconsistent within parity groups across the two experiments.