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.

Fertility in dairy cows after artificial insemination using sex-sorted sperm or conventional semen.

The aim of this study was to compare pregnancy per artificial insemination (P/AI) after timed AI with sex-sorted sperm (SS) or conventional semen (CS) in lactating dairy cows. Cyclic cows (n = 302) were synchronized by Ovsynch and randomly assigned into two groups at the time of AI. Cows with a follicle size between 12 and 18 mm and clear vaginal discharge at the time of AI were inseminated with either frozen-thawed SS (n = 148) or CS (n = 154) of the same bull. A shallow uterine insemination was performed into the uterine horn ipsilateral to the side of probable impending ovulation. Pregnancy per AI on Day 31 tended (p = 0.09) to be less for SS (31.8%) than CS (40.9%). Similarly, P/AI on Day 62 was less (p = 0.01) for cows inseminated with SS (25.7%) compared with CS (39.0%). The increased difference in fertility between treatments from Days 31 to 62 was caused by the greater (p = 0.02) pregnancy loss for cows receiving SS (19.2%) than CS (4.8%). Cow parity (p = 0.02) and season (p < 0.01) when AI was performed were additional factors affecting fertility. Primiparous cows had greater P/AI than multiparous cows both on Day 31 (41.7% vs 25.0% in SS and 53.0% vs 31.8% in CS groups) and on Day 62 (33.3% vs 20.5% in SS and 48.5% vs 31.8% in CS groups). During the hot season of the year, P/AI on Day 31 was reduced (p = 0.01) in the SS group (19.6%) when compared with the rates during the cool season (38.1%). In conclusion, sex-sorted sperm produced lower fertility results compared to conventional semen even after using some selection criteria to select most fertile cows.

Perosomus elumbis in a Holstein calf infected with bovine viral diarrhea virus.

The detection of bovine viral diarrhea virus (BVDV) in a female Holstein calf presented with perosomus elumbis, a congenital anomaly, is reported here. A cow with dystocia was evaluated and an abnormal dead calf was detected during vaginal examination. The calf was retrieved via caesarean section and exhibited abnormalities characteristic of PE, such as vertebral and pelvic malformations. These abnormalities were further confirmed using radiographic and necropsy examinations. At necropsy cerebellar hypoplasia was an additional finding, which is a typical lesion associated with bovine virus diarrhea (BVD). Several tissue samples from the calf were tested for the presence of antigens of BVDV and bovine herpesvirus-1 (BHV-1) by ELISA. In addition, sera samples from the dam and calf were tested for the presence of antibodies against BVDV, BHV-1, and bluetongue disease virus (BTV) using a virus neutralization assay. Results indicated that the calf was congenitally infected with BVDV, whereas there was no evidence for the presence of BHV-1 and BTV. In the dam's serum no antibodies against BVDV, BHV-1, and BTV were detected. Even though the etiology of perosomus elumbis is unknown, BVDV, which causes fetal anomalies at early gestation in cows, may have been a contributing factor in this case.

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.

Dry period management and optimization of post-partum reproductive management in dairy cattle.

Dry period and early post-partum management are decisive factors for fertility in lactating dairy cows. Previous studies have shown that decreased dry matter intake (DMI) and increased non-esterified fatty acids (NEFA) negatively affect fertility and subsequent milk production. The traditional dry period decreases DMI prior to parturition, resulting in a decrease in energy intake. A negative energy balance increases NEFA concentration, and increased NEFA may impair the immune system, especially by decreasing neutrophil function prior to parturition. Earlier studies have shown that post-partum health disorders, including retained placenta and metritis, were correlated with periparturient neutrophil function. In addition, decreased DMI is also linked to a reduced body condition score (BCS) in dairy cows. These events in the periparturient period negatively affect fertility. Some manipulation, such as shortening the dry period, may be a solution to increased DMI in the periparturient period, preventing post-partum disorders and subsequent fertility issues. This article aims to explain the effects of shortening the dry period on reproduction and early post-partum treatments to improve fertility. In addition, timed artificial insemination protocols will be discussed for use during the post-partum period to improve fertility in dairy cows.

Comparison of responses to Ovsynch between Holstein-Friesian and Swedish Red cows.

The Ovsynch protocol was designed to synchronize ovulation, thereby allowing timed artificial insemination (TAI) of all cows without detection of estrus. However, the effectiveness of Ovsynch in different breeds of dairy cows has not been previously compared. The aim of this study was to compare the response to Ovsynch in cycling lactating Holstein-Friesian (HF) and Swedish Red (SR) dairy cows. A total of 495 cyclic cows (n=347 HF, n=148 SR) were housed together and treated with Ovsynch (GnRH-7 d-PGF2α-56 h-GnRH-16 to 18 h-TAI). Ovulatory responses, synchronization rate, maximal follicle size at the time of AI, and percentage of pregnant cows per AI (P/AI at 31 and 62 d after AI) were compared between breeds. Ultrasonography was performed during Ovsynch at first GnRH, PGF2α, at time of AI, and 7 d after AI. Ovulatory response and synchronization rate were similar in HF versus SR cows (60.2 vs. 62.2%; 88.4 vs. 88.5%, respectively). Cows that ovulated to the first GnRH of Ovsynch had smaller follicle size at AI (15.9±0.1 vs. 16.4±0.2 mm). Maximal follicle size at AI was greater for HF (16.4±2.2 mm) than SR (15.5±2.3 mm) cows. The P/AI was greater for SR than HF cows at the 62-d pregnancy diagnosis (56.1 vs. 46.1%). In addition, pregnancy loss between 31 and 62 d of pregnancy was greater in HF (10.1%) than SR (3.5%) cows. Fertility was less in HF cows during the hot season (57.7 in cold vs. 38.1% in the hot season), whereas such a decrease was not observed in SR (60.0 in cold vs. 53.5% in the hot season) cows. Thus, although the GnRH treatments of Ovsynch were equally effective in SR and HF cows, pregnancy outcomes (P/AI at d 62 and pregnancy survival) were greater in SR than HF cows, and P/AI in SR cows was not compromised during the hot season as was found for HF cows.

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.

Managing the dominant follicle in high-producing dairy cows.

Reduced reproductive efficiency has been reported in high-producing dairy cows. Sources of reproductive inefficiency include decreased expression of estrus, increased diameter of the ovulatory follicle and reduced fertility when cows are inseminated after estrus, increased incidence of double ovulation and twinning, and increased pregnancy loss. To overcome some of these inefficiencies, reproductive management programs have been developed that synchronize ovulation and enable effective timed artificial insemination (AI) of lactating dairy cows. Effective regulation of the corpus luteum (CL), follicles, and hormonal environment are critical for optimizing these programs. Recent programs, such as the 5-day CIDR program, Double-Ovsynch, G-6-G, and estradiol benzoate-CIDR programs were designed to more effectively control one or more physiological events. These events include synchronization of a new follicular wave at the beginning of the program, optimization of the circulating progesterone (P4) concentrations and duration of follicular dominance, optimized reductions in P4 and increases in circulating estradiol (E2) concentrations during the preovulatory period, and tightly synchronized ovulation of a follicle of optimal size and fertility for implementation of timed AI. The success of these programs has been remarkable, although there is substantial variability in effectiveness due to environmental, management, nutritional, genetic, and disease factors as well as potential variability in some aspects of reproductive physiology among commercial dairy farms. Future programs will optimize the reproductive physiology while simplifying the protocol implementation and also match specific reproductive management protocols to specific farms and even specific cows (for example primiparous vs. multiparous).

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.

Reducing dry period length to simplify feeding transition cows: milk production, energy balance, and metabolic profiles.

Sixty-five Holstein cows were used to evaluate management schemes involving altered dry period (DP) lengths on subsequent milk production, energy balance (EB), and metabolic variables. Cows were assigned to one of 3 treatments: traditional 56-d DP (fed a low-energy diet from -56 to -29 d and a moderate energy diet from -28 d to parturition; T), 28-d DP (continuously fed a high energy diet; S), and no planned DP (continuously fed a high energy diet; N). Prepartum DM intake (DMI), measured from 56 d prepartum through parturition, was lower for cows on the T treatment than for cows on the S treatment and was higher for cows on the N treatment than for cows on the S treatment. There were no differences in prepartum plasma glucose, and beta-hydroxybutryric acid; there was a treatment by time interaction for prepartum plasma nonesterified fatty acid (NEFA). There was no difference in prepartum liver triglyceride (TG); postpartum liver TG was decreased for cows on the N treatment compared with cows on the S treatment, but was similar for cows on the T and S treatments. Postpartum NEFA was similar between cows on the T and S treatments, but was greater for cows on the S treatment than for cows on the N treatment. Postpartum glucose was greater for cows on the N treatment compared with cows on the S treatment and tended to be greater for cows on the S treatment than for cows on the T treatment. There was no difference in postpartum solids-corrected milk (SCM) production or DMI by cows on the T vs. S treatment. However, there was a tendency toward lower postpartum SCM production by cows on the N vs. S treatment and a tendency for greater postpartum DMI by cows on the N vs. S treatment. Postpartum EB was greater for cows on the S vs. T treatment and the N vs. S treatment. In general, T and S management schemes had similar effects on DMI, SCM, and metabolic variables in the first 70 d of the subsequent lactation. Eliminating the DP improved energy and metabolic status.

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.