This study examined the effect of a single administration of cephapirin iu or cloprostenol im on the reproductive performance of dairy cows with subclinical endometritis. Cows (n=228) at 20–33 days in milk (DIM) from two commercial dairy farms, determined to be normal for clinical endometritis (based on absence of abnormal uterine discharge on vaginoscopic examination) were enrolled. At enrollment, a thorough reproductive examination was performed, including rectal palpation, ultrasonography (US) and endometrial cytology (EC). The case definition for subclinical endometritis was the presence of >18% neutrophils on EC examination or fluid in uterus (FIU) on US examination. All cows were randomly assigned to receive one of three treatments: 500 mg benzathine cephapirin iu, 500 μg cloprostenol im, or control (no treatment). Reproductive performance was monitored for a minimum of 8 months after treatment. Cows with subclinical endometritis treated with cephapirin or cloprostenol had a significantly increased relative pregnancy rate compared to control [hazard ratios 1.89 (P=0.01) and 1.70 (P=0.05), respectively]. In conclusion, a single treatment with cephapirin or cloprostenol at 20–33 DIM significantly improved the reproductive performance of cows with subclinical endometritis.

This study compares cytobrush and lavage techniques for the assessment of endometrial cytology (EC) in clinically normal postpartum dairy cows. The EC samples were collected from Holstein cows (n = 35) during visit 1 (V1) at 20 to 33 d in milk (DIM) and 2 wk later during visit 2 (V2) at 34 to 47 DIM by using both techniques. A minimum of 100 cells were counted to determine the percentage of cells that were neutrophils (%PMN). The mean %PMN was significantly different between the techniques at V1 (P = 0.001), but not at V2 (P = 0.474). Overall, the %PM N decreased with time postpartum (r2 = 0.36; P = 0.001), but not within V1 (P > 0.05) or V2 (P > 0.1). Uterine diameter was negatively correlated with fluid recovery by the lavage technique (r2 = 0.41; P = 0.002). The mean %PMN was not influenced by the volume of fluid recovered in successful attempts, but 17% (12/70) of attempts yielded no fluid. In conclusion, the cytobrush technique is a consistent and reliable method for obtaining endometrial samples for cystologic examination from postpartum dairy cows.

Poor reproductive outcome associated with chromosome anomalies is well documented in humans and domestic animals. In cattle, female carriers of Robertsonian and X-autosome translocations tend to be repeat breeders, probably due to synaptic difficulties during meiotic prophase of gametogenesis. Although viable offspring have been obtained through somatic cell nuclear transfer (NT) using adult or fetal cells from normal animals in various species, there have been no reports to date on the application of this technology to translocation carriers. Since NT circumvents meiotic problems encountered by translocation carriers, we used this approach to generate cloned embryos, fetuses and calves from a subfertile Limousin-Jersey crossbred cow previously identified as a carrier of an X-autosome translocation. Primary cultures were established from ear skin biopsies, and used at the 5th or 6th passage for NT. Recipient oocytes were enucleated at 19h post-maturation (hpm), fused with individual fibroblasts by a single electrical pulse (1.4KVcm−1, 40μs) and activated at 24hpm with a combination of ionomycin (5μM, 5min) and cycloheximide (10μgmL−1). Reconstructed eggs were cultured in SOF at 39°C in a humidified low oxygen atmosphere. In 33 runs involving 2470 oocyte-donor cell complexes, cleavage and blastocyst rates were 88% (2173/2470) and 36% (889/2470), respectively. Two or three blastocysts (Day 7±1) were transferred into each recipient, previously synchronized with a combination of CIDR, GnRH and PGF2α. Ultrasonography was performed at Days 28 to 60 and at Days 90 and 150. Pregnancy was confirmed on Day 28 in 10 of a total of 22 recipients, 2 of which were later found to be carrying twin fetuses. Of 60 embryos transferred, 11 (18.3% of embryos) survived to Day 42, 6 (10%) to Day 60, and 4 (6.6%) to Day 90. A Day-94 fetus was surgically retrieved to examine the synaptic pattern of meiocytes in fetal ovaries. The fetus and internal organs were normal in appearance, and of normal size (16.5cm C-R length). The X-autosome translocation was confirmed in blood cultures, and synaptic anomalies involving chromosome 23 and the X chromosome were detected in fetal ovaries. Another clone was delivered by C-section at 276 days but died within 1h of delivery, while one singleton pregnancy is still ongoing at >200 days. These results demonstrate that NT can be used to produce embryos, fetuses and offspring from an X-autosome translocation carrier, with the potential to facilitate study of synaptic behaviour of female germ cells, and X-inactivation in different cell lineages of cloned blastocysts, and to generate individuals with otherwise poor reproductive prospects. [Supported by NSERC, Canada and OMAFRA]

The objective of this research was to determine if PGF2alpha-induced milk letdown (ML) is an accurate indicator of luteolysis, allowing cows to be synchronized to begin the Ovsynch protocol (GnRH-7d-PGF2alpha-2d-GnRH-24h-AI) at the most beneficial time of the estrous cycle (days 5-9), and determine if this would improve pregnancy rate (PR). Lactating Holstein cows between 55 and 70 days in milk were used to evaluate the ML test and PR after the Ovsynch protocol, when initiated on the basis of the test result (PROSYNCH). PROSYNCH cows (n = 60) had one teat cannulated to test for ML and were treated with 500 microg cloprostenol, PGF2alpha analogue (PG). Cows with ML were started on Ovsynch 10 days later, and those without started 3 days later. Cows in the control group (OVSYNCH, n = 64) were injected with physiological saline and observed for ML. This group was started on Ovsynch 10 days after saline treatment. Milk samples were collected thrice weekly to determine progesterone concentrations. ML indicated luteolysis with a sensitivity of 98% and a specificity of 60%. The positive and negative predictive values were 83 and 92%, respectively. Pregnancy rates were 48% for PROSYNCH and 52% for OVSYNCH (P = 0.72). When data from both groups were combined, PR was greater in cows that started the Ovsynch protocol in stage 2 of the estrous cycle (days 5-9, 67%) than all other stages (stage 1: days 1-4, 35%; stage 3: days 10-16, 45%; stage 4: days 17-21, 42%; P < 0.01). The proportion of animals with ovulation after GnRH#1, luteolysis after PGF2alpha, and ovulation after GnRH#2 were all greater in the PROSYNCH group (77% versus 55%, P < 0.02; 83% versus 66%, P < 0.03; 97% versus 84%, P < 0.03, respectively). Therefore, the ML test indicated luteolysis with sufficient precision to time the initiation of the Ovsynch protocol between days 5 and 9 of the cycle, however, this did not alter PR compared to starting the protocol randomly throughout the cycle. Initiating the Ovsynch protocol between days 5 and 9 of the cycle increased PR, and improved the efficacy of each injection.

The objectives of the study were to validate the use of endometrial cytology (EC) and ultrasonography (US) to diagnose subclinical endometritis in clinically normal postpartum dairy cows, and to measure the impact of subclinical endometritis on reproductive performance. Holstein cows from two dairy farms were examined at Visit 1 (V1) at 20–33 days in milk (DIM), and clinically normal cows (n=228), based on the absence of abnormal discharge on external inspection and vaginoscopy, were selected. The reproductive tract of selected cows was evaluated by transrectal palpation, US and EC. All cows in the study were re-examined at Visit 2 (V2) at 34–47 DIM (2 weeks after V1) and were subsequently followed for a minimum of 8months (until pregnant or culled). Survival analysis was used to derive a case definition of subclinical endometritis, based on factors associated with decreased relative pregnancy rate. Positive EC at V1 (>18% polymorphonuclear leukocytes; PMN) or fluid in uterus at V1 (FIU1) were associated with a significant reduction in the relative pregnancy rate and identified cows with subclinical endometritis. Similarly, a positive EC (>10% PMN) at V2 or fluid in the uterus at V2 (FIU2), identified cows with subclinical endometritis. Cows with subclinical endometritis at V1 and at V2 had a relative pregnancy rate of 41 and 51% (hazard ratio for pregnancy of 0.59 and 0.49), respectively, compared to cows without subclinical endometritis. Given EC or US findings, no diagnostic criteria based on transrectal palpation of the uterus had predictive value for risk of pregnancy. In conclusion, subclinical endometritis, diagnosed by EC or US, was associated with reduced relative pregnancy rate.

The differences in the embryo production potential of four rams used in a commercial embryo transfer program were examined in both in vivo and in vitro embryo production systems. Processing frozen-thawed spermatozoa through Percoll density gradients prior to in vitro insemination eliminated differences in the estimates of sperm viability between the four rams, and yet, differences in embryo production persisted throughout the in vitro culture period. However, there was no effect of ejaculate within ram on embryo production rates. In addition, the timing of the onset of the differences between the rams at each stage of in vitro embryo development were revealed. Ram 2 differed from ram 4 in the proportion of fertilized oocytes at 17 h post-insemination (pi) and by 52 h, ram 3 differed from ram 4 in the proportion of cleaved embryos, and the observed differences between ram 1 and ram 2 in their blastocyst production were initiated prior to activation of the embryonic genome. Once differences in embryo development rates were detected among the four rams, they persisted throughout the in vitro culture period. The reduced in vitro fertilization (IVF) rates from ram 2 compared with the other rams was paralleled in vivo by the significantly lower proportion of embryos recovered from ewes mated to ram 2, and this was further exacerbated by a significantly lower embryo survival rate after transfer. However, the subtle differences observed in the timing of the contribution of each sire to embryo development during in vitro culture were not able to be detected in vivo. However, the higher proportions of transferable quality blastocysts obtained from ewes mated to ram 4 did not result in increased embryo survival throughout the remainder of gestation. Therefore, in this study, the blastocyst production potential for a particular sire, either in vitro or in vivo, does not necessarily reflect the potential for the production of live offspring.

The objective of this study was to investigate parenteral vitamin E for the prevention of peripartum disease in dairy cows. A randomized clinical trial was conducted in 21 commercial dairy herds. Cows (n = 1142) were randomly assigned to receive either a single subcutaneous injection of 3000 IU of vitamin E, or placebo, 1 wk before expected calving. Serum alpha-tocopherol was significantly increased in treated cows at 7 and 14 d, but not at 21 d after injection. Overall, there were no significant differences between treatment groups in the incidence of retained placenta, clinical mastitis, metritis, endometritis, ketosis, displaced abomasum, or lameness. However, there was a conditional benefit of treatment for reduction of the incidence of retained placenta. Cows with marginal pretreatment vitamin E status (serum alpha-tocopherol to cholesterol mass ratio < 2.5 x 10(-3)) that received an injection of vitamin E tended to have reduced risk of retained placenta. However, in cows with adequate serum vitamin E, there was no reduction in the incidence of any disease. For clinical application, primiparous animals were most likely to benefit from prepartum injection of vitamin E.

The objective of this field trial was to compare the effect of intrauterine (i.u.) antibiotic or intramuscular (i.m.) prostaglandin F2 alpha (PGF2 alpha) on time to pregnancy in dairy cows diagnosed with clinical endometritis between 20 and 33 days in milk (DIM). The case definition of endometritis was the presence of purulent uterine discharge or cervical diameter > 7.5 cm, or the presence of muco-purulent discharge after 26 DIM. There were 316 cows with endometritis from 27 farms assigned randomly within herd to receive 500 mg of cephapirin benzathine intrauterine (i.u.), 500 micrograms of cloprostenol i.m., or no treatment. The rate of resolution of clinical signs 14 d after treatment was 77% and was not affected by treatment. Reproductive performance was monitored for a minimum of 7 mo after treatment. Survival analysis (multivariable proportional hazards regression) was used to measure the effect of treatment on time to pregnancy. There was no benefit of treatment of endometritis before 4 wk postpartum. Administration of PGF2 alpha between 20 and 26 DIM to cows with endometritis that did not have a palpable corpus luteum was associated with a significant reduction in pregnancy rate. Between 27 and 33 DIM, cows with endometritis treated with cephapirin i.u. had a significantly shorter time to pregnancy than untreated cows (hazard ratio = 1.63). In this time period, there was no difference in pregnancy rate between PGF2 alpha and untreated cows, but the difference in pregnancy rate between cows treated with cephapirin i.u. and with PGF2 alpha was not statistically significant. Treatment of postpartum endometritis should be reserved for cases diagnosed after 26 DIM, based on criteria that are associated with subsequent pregnancy rate.

The objectives of this study were to validate diagnostic criteria for clinical endometritis in postpartum dairy cows and to measure the impact of endometritis on reproductive performance. Data were collected from 1865 cows in 27 herds, including history of dystocia, twins, retained placenta, or metritis. All cows were examined once between 20 and 33 d in milk (DIM) including external inspection, vaginoscopy, and transrectal palpation of the cervix, uterus, and ovaries. All cows were followed for a minimum of 7 mo or until pregnancy or culling. Survival analysis was used to derive a case definition of endometritis based on factors associated with increased time to pregnancy. The significance of clinical findings depended on the interval postpartum when examination took place. The presence of purulent uterine discharge or cervical diameter > 7.5 cm after 20 DIM, or mucopurulent discharge after 26 DIM identified cows with clinical endometritis. Given vaginoscopy, no diagnostic criteria based on palpation of the uterus had predictive value for time to pregnancy. The prevalence of clinical endometritis was 16.9%. Vaginoscopy was required to identify 44% of these cases. Accounting for parity, herd, and ovarian status, cows with clinical endometritis between 20 and 33 DIM had a hazard ratio of 0.73 for pregnancy (took 27% longer to become pregnant), and were 1.7 times more likely to be culled for reproductive failure than cows without endometritis.

To select rams suitable for ovine in vitro embryo production (IVP), the predictive values of the screening tests used to identify unsuitable rams need to be established. The present study examined some characteristics of frozen-thawed ram spermatozoa that might be evaluated routinely in a commercial breeding programme. These included sperm motility, plasma membrane integrity, morphology, and acrosome and capacitation status of the sperm population. Cryopreserved spermatozoa from four Dorset rams, which had previously satisfied the selection criteria for inclusion in a commercial breeding programme, were used for IVP. The overall contribution of the four rams and the ejaculates within each ram to the variability (R2) in the production of blastocysts was very small (2.1% and 2.5% respectively). The analysis of the sperm characteristics by logistic regression revealed a significant and positive association between total post-thaw sperm motility, viability and longevity with in vitro blastocyst production. However, there was no association between the other surface characteristics of the spermatozoa measured in this study with embryo production. Despite the absence of differences between the rams in the low incidence of polyspermic fertilization, the significant and detrimental effects of polyspermic fertilization on in vitro blastocyst production rates were quantified by logistic regression analysis. A large proportion of the variability within the IVP system was unaccounted for by the analysis of sperm and oocyte characteristics evaluated in this study. Thus, the identification of other factors contributing to the variability in the production of embryos in vitro warrants further investigation. No single sperm characteristic was sufficient to predict the ultimate outcome of blastocyst production. Rather, assessments of multiple characteristics within the IVP system are required to make accurate predictions.

Lactating Holstein cows (n=288) were grouped as pairs at parturition and randomly assigned to two treatments (control, C vs intervenient treatment, T). The reproductive management of the Group C cows (n=130) consisted of the intramuscular administration of 500 microg PGF2alpha analogue (PG) on Days 28 and 63 postpartum and breeding on the basis of estrus signs with the a.m.-p.m. rule after Day 63. Cows that were not bred by 77 d postpartum received another injection of PG and were bred at estrus or 84 h after PG treatment. Pregnancy diagnoses were perfomed by palpation of the uterus per rectum 42 to 48 d after AI. Cows in the T group (n=139) received intramuscular injections of 100 microg GnRH 14 d and PG 28 d after calving. On Day 56 postpartum, cows were given a second dose of GnRH followed by PG on Day 63 postpartum and a third GnRH injection 48 h after PG (OvSynch). Cows were inseminated at a fixed time (22+/-1 h) after GnRH. Five days after the fixed-time insemination cows were given 1500 IU hCG i.m.. Group C and T cows that returned to service or were diagnosed as non-pregnant continued to receive PG at intervals of 14 d with breeding at estrus or 84 h after the second PGF2alpha dose. A sustained increase in milk progesterone concentration was observed in 59.0% of T cows after GnRH administration on Day 14. A similar rise in milk progesterone concentrations was observed in 53.8% of C cows. The PG on Day 28 induced luteolysis more in Group T cows (53.2%) than in Group C cows (36.9%). The PG on Day 63 reduced milk progesterone concentrations to basal levels in 50.7% of T and 49.2% of Group C animals. The first service pregnancy rates (T, 40.3% vs C, 36.2%) and the overall pregnancy rates (all services, T, 83.5% vs C, 86.9%) were not different between the two groups. The two treatments did not differ in the interval from first service to pregnancy, calving to pregnancy or in calving interval, number of services per pregnancy or culling rates.

Holstein heifers were used to study effects of exogenous administration of oxytocin on luteal function and ovarian follicular development. Twelve heifers were monitored for 1 estrous cycle to confirm normal ovarian function. At the subsequent estrus, these animals were randomly assigned to 1 of 3 treatments: saline control, (Group 1, n = 4), oxytocin (Group 2, n = 4) and saline pregnant (Group 3, n = 4). Group 2 received continuous infusion of oxytocin (1.9 mg/d) from Days 14 to 26 after estrus, while Groups 1 and 3 received saline infusion during the same period. Group 3 were artificially inseminated at estrus. Daily blood samples were collected for oxytocin and progesterone assay. Ovarian follicles and corpus luteum (CL) development were monitored daily by transrectal ultrasonography until Day 32 after estrus. Plasma progesterone (P4) concentrations prior to initiation of infusion were 7.6 ± 1.3 ng/mL on Day 14. They then decreased to <1 ng/mL on Day 19 for Group 1 and on Day 28 for Group 2. The interestrous interval was longer (P<0.05) for heifers that received oxytocin infusion. During the infusion period P4 concentrations were not different (P >0.05) between Group 2 and 3 but declined gradually from Day 20 in Group 2 despite the presence of high plasma oxytocin concentrations. Control heifers had 2 waves of follicular growth, with the second dominant follicle ovulating. Three of the 4 oxytocin-infused animals had an additional wave, with the third dominant follicle ovulating. Oxytocin infusion had no effect on size of the ovulating follicle (P>0.05) and the number of Class 1 follicles (3 to 5 mm, P>0.1). Differences in the number of Class 2 follicles (6 to 9 mm) among treatments on Days 15 to 22 after estrus were not detected (P>0.1) except on Days 23 to 26, when Group 2 had fewer follicles than Group 3 (P <0.05).The results show that continuous infusion of oxytocin during normal luteolysis delays luteal regression without inhibiting follicular development.

The profitability of beef breeding farms in New Zealand depends principally on optimal reproductive performance. The aim of this study was to estimate the impact of four major pathogens, bovine viral diarrhea virus (BVDV), Neospora caninum (N. caninum), Leptospira borgpetersenii serovar Hardjo (Hardjo), and Leptospira interrogans serovar Pomona (Pomona), on rates of fetal loss in commercial beef breeding herds. Farms reporting fetal loss were recruited, and a blood sample from aborting cows (cases) was collected. Controls were normally calving cows from the same farm. At least four controls were selected from each farm contributing cases. Samples were tested using ELISA for detection of antibodies against BVDV and N. caninum, and microscopic agglutination test (MAT) for detection of antibody against Hardjo and Pomona. A selection of titer cut-offs was conducted to evaluate the relationship between fetal loss and seropositivity to each pathogen using conditional logistic regression. The cut-off titer with the strongest association with fetal loss was included in the multivariate model. A significant increased risk of fetal loss was found for animals seropositive to N. caninum (odds ratio (OR) = 3.36; 95% confidence interval (95% CI) = 1.27–8.89), Hardjo (OR = 1.84; 95% CI = 1.01–3.33), and Pomona in non-vaccinated cows (OR = 14.91, 95% CI = 1.73–128.84) at the ELISA titer ≥30, and MAT titers of ≥1:384 and ≥1:768 for a positive sample, respectively. A marginally non-significant increased risk of fetal loss was found for animals exposed to BVDV (OR = 2.01; 95% CI = 0.99–4.11) at the ELISA titer of ≤1. Vaccination did not affect ORs for Hardjo or BVDV and no herd vaccinated against N. caninum. Approximately 14.0% of all fetal loss in the beef breeding cattle population in New Zealand may be attributable to BVDV (3.5%), N. caninum (3.0%), Hardjo (4.7%), and Pomona (3.6%).

This study was designed to evaluate the amounts of coagulation factors and to determine whether the protein profile in pre-ovulatory ovarian follicular fluid aspirated from ovaries collected from mares at slaughter are representative of that in follicular fluid collected from live animals. The proteins evaluated included, (i) albumin, ceruloplasmin and fibronectin, (ii) the procoagulant plasma proteins, Factor V (FV), Factor VII (FVII), Factor X (FX) and prothrombin, and (iii) the anticoagulant plasma proteins, antithrombin and alpha2-macroglobulin. The amounts of the individual proteins were similar in both types of follicular fluid. There was no correlation between the activity of FV, FVII, FX or prothrombin in follicular fluid and their molecular size although a correlation was found for the other proteins. These results suggest that the procoagulant proteins in follicular fluid are not likely derived from plasma. The total protein content of follicular fluid samples collected from both sources was similar and the results determined with the Biuret, Lowry and Biorad methods were also not significantly different (P>0.05).

Twenty-five normally cyclic Holstein heifers were used to examine the effects of oxytocin on cloprostenol-induced luteolysis, subsequent ovulation, and early luteal and follicular development. The heifers were randomly assigned to 1 of 4 treatments: Group SC-SC (n=6), Group SC-OT (n=6), Group OT-SC (n=6) and Group OT-OT (n=7). The SC-SC and SC-OT groups received continuous saline infusion, while Groups OT-SC and OT-OT received continuous oxytocin infusion (1:9 mg/d) on Days 14 to 26 after estrus. All animals received 500 microg, i.m. cloprostenol 2 d after initiation of infusion (Day 16) to induce luteolysis. Groups SC-OT and OT-OT received oxytocin twice daily (12 h apart) (0.33 USP units/kg body weight, s.c.) on Days 3 to 6 of the estrous cycle following cloprostenol-induced luteolysis, while Groups SC-SC and OT-SC received an equivalent volume of saline. Daily plasma progesterone (P4) concentrations prior to cloprostenol-induced luteolysis and rates of decline in P4 following the induced luteolysis did not differ between oxytocin-infused (OT-OT and OT-SC) and saline-infused (SC-SC and SC-OT) groups (P >0.1). Duration of the estrous cycle was shortened in saline-infused heifers receiving oxytocin daily during the first week of the estrous cycle. In contrast, oxytocin injections did not result in premature inhibition of luteal function and return to estrus in heifers that received oxytocin infusion (OT-OT). Day of ovulation, size of ovulating follicle and time of peak LH after cloprostenol administration for oxytocin and saline-treated control heifers did not differ (P >0.1). During the first 3 d of the estrous cycle following luteal regression, fewer (P <0.01) follicles of all classes were observed in the oxytocin-infused animals. Day of emergence of the first follicular wave in heifers treated with oxytocin was delayed (P <0.05). The results show that continuous infusion of oxytocin during the mid-luteal stage of the estrous cycle has no effect on cloprostenol-induced luteal regression, timing of preovulatory LH peak or ovulation. Further, the finding support that an episodic rather than continuous administration of oxytocin during the first week of the estrous cycle results in premature loss of luteal function. The data suggest minor inhibitory effects of oxytocin on follicular growth during the first 3 d of the estrous cycle following cloprostenol-induced luteolysis.

Follicular growth and ovulation in response to FSH, progesterone and hCG were evaluated in postpartum beef cows. In Experiment 1, on Day 21 post partum, cows received an injection of either saline (control; n=6), FSH (200 mg; n=6), or a PRID (n=5) for 10 d. Both FSH and PRID prolonged maintenance of a dominant follicle (15.5 ± 1.16 and 14.4 ± 1.29 d, respectively, vs 8.4 ± 1.22 d in control; P < 0.01), and increased the maximum diameter of the dominant follicle (14.0 ± 0.91 and 16.4 ± 1.01 mm, respectively, vs 10.9 ± 0.95 mm in control; P < 0.05). The PRID-maintained dominant follicle ovulated in 60% of cows, followed by normal estrous cycles (vs 0% in control; P=0.01), whereas the dominant follicle ovulated in 33 % of FSH-treated cows (P=0.08). The PRID regimen shortened the interval to first ovulation preceding a normal cycle and continued cyclicity (44 ± 4.1 vs 60 ± 4.4 d in control; P=0.02). In Experiment 2, on Day 21 post partum, cows received either saline (control), saline+PRID, or FSH+PRID (n= 16/group). Sixty hours after PRID withdrawal, cows received either saline or hCG (1,500 IU, n=8/treatment). The FSH+PRID regimen increased the number of large (>10 mm in diameter) follicles (3.6 ± 0.43 vs 1.9 ± 0.39 in control; P=0.005). Both PRID and FSH+PRID prolonged maintenance of the largest follicle (11.0 ± 0.82 and 11.2 ± 0.91 d, respectively, vs 8.7 ± 0.81 d in control; P < 0.05). The PRID-maintained dominant follicle ovulated in 50% of cows, followed by normal estrous cycles. The FSH+PRID-maintained largest follicle had become atretic at PRID withdrawal and was anovulatory. The FSH+PRID+hCG regimen increased the incidence of ovulation preceding a cycle of normal duration and continued cyclicity (100 vs 50% in PRID; P=0.03), and reduced the interval to first ovulation preceding a cycle of normal duration and continued cyclicity (38 ± 6.5 vs 58 ± 6.3 d in control; P=0.04). The area under the progesterone curve during the induced cycle was reduced after (PRID ± FSH)+hCG than after PRID ± FSH (P=0.002). These results indicate that PRID alone or with FSH/hCG has the potential to modify the dominant follicle and initiate cyclicity in postpartum beef cows.

The objectives of this study were to evaluate ovarian changes in cows with follicular ovarian cysts following treatment with either GnRH or saline. The parameters determined were the intervals from treatment to observation of a CL and from treatment to disappearance of the cyst, and the association between serum concentrations of LK, FSK and the LH/FSH ratio, before and after treatment, with the test intervals. Thirty-nine cows were identified as having follicular cysts. The GnRH treatment induced a significant increase in LH and the LH/FSH ratio. The gonadotropin response was not associated with the intervals from treatment to CL detection or to disappearance of the cyst. Survival curves for the intervals from treatment to CL detection and cyst disappearance indicate that treatment with GnRH or saline did not yield significantly different results for either parameter. The results question the efficacy of treating cystic ovarian disease with GnRH. (C) 1999 by Elsevier Science inc.

Segmental aplasia of the left uterine horn in a multiparous Holstein cow was diagnosed by palpation and ultrasonography. Treatment with prostaglandin was unsuccessful in eliminating the fluid from the distended uterine horn. Segmental aplasia should be included in the list of differential diagnoses for cows with nonresponsive uterine enlargement.

This study assessed pronuclear formation, the chromosomal constitution, and the developmental capacity of bovine zygotes formed by intracytoplasmic injection of oocytes with sperm, treated or not with dithiothreitol (DTT). Oocytes were matured in vitro for 22-24 h and then centrifuged so that sperm, prepared by swim-up in the presence or absence of 5 mM DTT, could be injected into the cleared area of the ooplasm. Injected oocytes were activated by treatment with 5 microM ionomycin (5 min) and, after a 3-h interval, with 1.9 mM 6-dimethylaminopurine (DMAP) for 3 h. They were then cocultured with bovine oviductal epithelial cells in M199. Sperm treatment resulted in a significantly higher proportion of male pronucleus formation 16 h after injection (40% vs. 11%; p < 0.0001) and a significantly higher rate of blastocyst development (24% vs. 10%; p < 0.005). Sixty-one percent of blastocysts produced with treated sperm were diploid. Of 12 blastocysts produced with treated sperm and sexed by a polymerase chain reaction, 4 were male and 7 female, and in one a definite diagnosis could not be made. Embryo transfer (2 embryos per heifer) resulted in pregnancies in 6 of 16 recipients at Day 49, but none was carried to term. These results show that the efficiency of bovine intracytoplasmic sperm injection can be improved by sperm pretreatment with DTT and by oocyte activation with ionomycin plus DMAP, although the developmental capacity of the resulting embryos remains limited.