Early pregnancy diagnosis by transrectal ultrasonography in ewes

Two studies were performed to evaluate early pregnancy diagnosis by transrectal ultrasonography (TRUS), using a 7.5-MHz transducer in ewes. In the first study, the objectives were to determine the earliest date that a reliable pregnancy diagnosis could be made (percentage of ewes detected pregnant between days 15 and 20 after mating). In the second study, the objective was to confirm the findings of the first study using a randomized controlled trial. In both studies, the ewes were restrained in dorsal recumbency, using a special chute that maintained the pelvis at an angle of 30–35° lower than the head. In the first study, 30 Suffolk ewes were synchronized and maintained with 2 rams for 5 days. Each ewe was subjected to the first TRUS on day 15 after mating and daily thereafter until day 20 (estrus = day 0). Pregnancy was defined as the presence of an embryo or extra-embryonic membranes. The percentage of ewes detected pregnant at days 15, 16, 17, 18, 19 and 20 were 0% (0/30), 26.7% (8/30), 86% (24/30), 90% (27/30), 96.7% (29/30) and 100% (30/30), respectively (P < 0.001). In the second study, 390 TRUS examinations (TRUS-1) were performed on ewes from 10 to 50 days after mating in a breeding program (group mating, hand mating, cervical and intrauterine AI; breeding group; n = 270) or with vasectomized rams (vasectomized group; n = 120). The breeding date and the status of breeding were unknown to the operator. Thirty of these ewes were mated with vasectomized rams and used repetitively four times as the non-pregnant control group. All females had a subsequent TRUS (TRUS-2) between 7 and 30 days after the TRUS-1 examination. The second TRUS was used as the standard test against which the performance of the TRUS-1 was compared. The percentage of ewes correctly diagnosed at day 15 or less, days 16, 17, 18, and 19 in the breeding group were 0% (0/29), 31.3% (5/16), 40% (8/20), 70% (7/10), and 100% (14/14), respectively (P < 0.001). All the diagnoses of ewes more than 20 days following mating in the breeding group, were correctly predicted (n = 181), as well as all ewes from the vasectomized group (n = 120). It could thus be concluded that the earliest pregnancy diagnosis using a 7.5-MHz transducer by transrectal route based on the presence of positive signs of pregnancy is at day 16 and the maximum sensitivity and negative predictive value was reached at day 20 following breeding.     

Early pregnancy diagnosis by palpation per rectum: influence on embryo/fetal viability in dairy cattle

The objective was to estimate the effect of palpation per rectum (for early pregnancy diagnosis) on embryo/fetal viability in dairy cattle. A controlled, randomized block-design experiment with two blocks, one by category, and the other by number of embryos, was conducted. Five-hundred-and-twenty pregnant dairy cows and heifers with a viable embryo detected by transrectal ultrasonography (TRUS) between days 29 and 32 after AI were included. The pregnant females were randomly allocated into two nearly equal groups: palpation per rectum (PAL group; n=258) and no palpation per rectum (NPAL group; n=262). The PAL group was submitted to palpation per rectum (PPR) using the fetal membrane slip (FMS) technique once between days 34 and 41 of pregnancy. The fetal membrane slip consisted of compressing the pregnant uterine horn and allowing the chorioallantoic membrane to slip between the fingers. Both groups were submitted to two additional TRUS at days 45 and 60 of pregnancy, to monitor the potential immediate and delayed deleterious effects of PPR on embryo and fetal viability, respectively. A diagnosis of embryo/fetal death was made when there was no embryo/fetal heart beat or the absence of positive signs of pregnancy in an animal previously diagnosed pregnant, or the presence of signs of embryo/fetal degeneration. The overall rate of embryo/fetal death was 14.0% (73/520). Embryonic death (10%; 52/520) was higher than fetal death (4.5%; 21/468; P<0.001). Embryo/fetal mortality was higher in cows (16.4%; 59/360) than in heifers (8.8%; 14/160; P<0.025) and in cattle with twin (25.5%; 12/47) versus singleton pregnancies (12.9%; 61/473; P<0.025), but was not different (P>0.05) between PAL (14.7%; 38/258) and NPAL (13.4%; 35/262). In conclusion, PPR between days 34 and 41 of pregnancy using the fetal membrane slip technique did not affect embryo/fetal viability.     

Effects of maturity of the potential ovulatory follicle on induction of oestrus and ovulation in cattle with oestradiol benzoate

The effect of maturity of the dominant follicle (DF) on the capacity of oestradiol benzoate (ODB) to induce oestrus and ovulation was examined in cattle. In experiment 1, 31 prepubertal heifers each received an intravaginal progesterone insert (IPI) and 1mg ODB i.m./500kg BW (ODB1). Daily ovarian ultrasonography detected emergence of a new follicular wave 3.1+/-0.1 days after ODB1. The IPI was removed when newly emerged DF were "young" (1.3+/-0.1 days after emergence; YDF; n=15) or "mature" (4.2+/-0.1 days; MDF; n=16), and 24h later, heifers received 0.75mg ODB/500kg BW (ODB2; n=16) or no further treatment (NoODB2; n=15). Most of the heifers receiving ODB2 were observed in oestrus (15/16) and ovulated (12/16), as compared to 0/15 and 1/15 in the NoODB2 group, respectively (P<0.01). In experiment 2, 32 heifers received ODB1 on day 6 of the oestrous cycle, and new follicular wave emergence was detected 3.2+/-0.1 days later. Heifers received an injection of prostaglandin-F2alpha (PGF) when the DF was young (1.1+/-0.1 days after emergence; YDF; n=16) or mature (4 days; MDF; n=16), and then ODB2 24h later or no further treatment (NoODB2). The interval from PGF to oestrus was greater (P<0.01) in the YDF-NoODB2 (70+/-3.9h) as compared to MDF-NoODB2 group (57+/-1.8h). Inclusion of ODB2 reduced (P<0.01) this interval to 47.0+/-0.7h without regard to the maturity of the DF (maturityxODB2, P<0.05) and also reduced (P<0.05) the interval to ovulation. In experiment 3, 21 suckling anoestrous cows received an IPI and ODB1 at 29.3+/-1.7 days postpartum. The IPI were removed either 1 day (YDF; n=9) or 3.9+/-0.1 days (MDF; n=9) after emergence of a new follicular wave and every cow received ODB2. Oestrus was subsequently detected in all but one animal. Ovulation of the newly emerged DF was detected within 48h of ODB2 in nine of nine cows of the MDF group, and in four of nine of the YDF group (P<0.05). During the subsequent ovulatory cycle, luteal size and plasma concentrations of progesterone were greater (P<0.01) in the MDF group compared to the YDF group. We conclude that behavioural oestrus is readily induced by 0.75mg ODB i.m./500kg BW. Maturity of the DF appeared to have little influence on the ability of the DF to ovulate in heifers. In contrast, young DF in lactating anoestrous cows were less likely to respond to the ovulatory cue provided, and luteal development was compromised in those that did ovulate.     

Intravaginal progesterone device and/or temporary weaning on reproductive performance of anestrous crossbred Angus x Nelore cows

The objective of this experiment was to evaluate effects of treatments with intravaginal progesterone (P4) device and/or 48h temporary weaning (TW) on reproductive performance of postpartum anestrous crossbred AngusxNelore cows, throughout the breeding season (BS). Anestrous cows (n=286; 53+/-5 DPP, body condition score between 2.5 and 3.5, on a 1-5 scale) were assigned randomly to four treatments (2x2 factorial design): (1) Control Group (no treatment; n=73); (2) TW Group (48h TW beginning on day 0 of BS; n=70); (3) P4 Group (Intravaginal P4 device between days -7 and 0 of BS; n=73); and (4) P4+TW Group (Intravaginal P4 device between days -7 and 0 of BS followed by 48h TW; n=70). On day 0 of BS (when P4 devices were removed and TW was initiated), cows had their ovaries evaluated by ultrasonography aiming to evaluate size of the largest follicle. Signs of behavioral estrus were observed twice a day (morning and evening), between days 0 and 25 of BS, and cows observed in estrus were inseminated 12h later. Between days 26 and 80 of BS, cows were submitted to natural service. Pregnancy diagnosis was conducted by transrectal ultrasonography on days 32 and 53 (pregnancies by AI) and 114 (pregnancies by natural service). Rates of detection of estrus during the first 3 days and within 25 days of BS were greater in cows receiving TW (45.0% compared with 9.6% and 50.0% compared with 29.5%, respectively; P<0.01). Conception rate in cows inseminated in the first 3 days of BS was greater in cows treated with P4 (54.8% compared with 11.1%; P<0.01). Combining P4+TW increased pregnancy rate in the first 3 days (Control: 1.4%, TW: 4.3%, P4: 8.2% and P4+TW: 24.3%; P<0.01) and in 25 days of BS (Control: 10.9%, TW: 7.1%, P4: 13.7% and P4+TW: 28.6%; P<0.05). Pregnancy rate at the end of BS did not differ among treatment groups. In conclusion, TW alone increased estrous behavior whereas P4 alone benefited conception. Combining P4 with TW improved pregnancy rate with direct benefits to behavioral estrus and conception.     

Embryo yield and plasma progesterone profiles in superovulated dairy cows and heifers.

This study was conducted to compare the superovulatory (SOV) response of dairy cows (n=172) and heifers (n=172), with two SOV treatments started at the mid-luteal-phase of the estrus cycle. Donors were randomly treated either with equine chorionic gonadotrophin (eCG) plus neutra-eCG serum (eCG+N group, n=167) or follicle stimulating gonadotrophin (FSH-P group, n=177).No significant differences were observed among groups in the percentage of superovulatory responsive donors (SR donors; corpora lutea (CL) >/=2), the mean number of total ova, fertilized ova and viable embryos recovered. Cows yielded significantly less total ova and less fertilized ova (P<0.05) and tended to yield less viable embryos (P<0.06) than heifers.Plasma progesterone (P4) concentrations (n=135 donors) on the day of PGF(2alpha) (PGF) injection and on the day of SOV estrus were significantly higher (P<0.01) in eCG+N than in FSH-P donors and, the increase between those 2 days was also significantly higher (P<0.05) in group eCG+N than in group FSH-P, suggesting a higher luteotrophic effect of eCG than FSH-P. SR donors had P4 levels significantly higher (P<0.001) than non-SR donors only on day 5 after the SOV estrus and on the day of embryo recovery. Plasma P4 concentrations at 5 days after the SOV estrus and at embryo recovery correlated significantly (r=0.76, P<0.001).Heifers had significantly higher P4 levels than cows at gonadotrophin injection (P<0.01), PGF injection (P<0.001), 5 days (P<0.01) and 7 days (P<0.001) after the SOV estrus. At day 7 after the SOV estrus, P4 concentrations per ova recovered were significantly higher in heifers than in cows (P<0.01). The increase of plasma P4 per ova recovered, between days 5 and 7 after the SOV estrus, was significantly (P<0.01) higher in heifers than in cows. Also, the increase of plasma P4 between injections of gonadotrophin and PGF was significantly higher (P<0.05) in heifers than in cows.These results suggest that heifers have higher plasma P4 concentrations at diestrus (either before or after the SOV treatment) and this is associated with a higher embryo yield and quality, as compared to lactating cows. These higher plasma P4 concentrations reflect not only differences in ovulation rate as well as the competence of the corpus luteum, which is potentialized by gonadotrophin stimulation.     

Effects of dietary energy level on luteinizing hormone and adrenal function in the post partum beef cow

The effects of dietary energy and suckling on adrenal function and luteinizing hormone (LH) concentrations were investigated in primiparous postpartum cows. Ten heifers were assigned at calving to either high (22.8 Mcal/day) or low (15.2 Mcal/day) energy diets. Blood samples were collected every 15 minutes for 8 hours on 28, 42, and 56 days post partum. Calves were allowed to suckle ad libitum during sampling periods. Serum samples were analyzed by radioimmunoassay for LH and cortisol. Concentrations of catecholamines were quantified by reverse-phase HPLC. Body weights were decreased (P<0.01) by low energy intake. In addition, low energy diet cows had lower mean LH concentrations (0.97 +/- 0.09 vs 1.57 +/- 0.07 ng/ml), P<0.05) than high energy diet cows. Luteinizing hormone concentrations in high energy diet cows increased with days post partum, resulting in a treatment-by-time interaction (P<0.005). Treatment did not affect mean cortisol concentrations. However, within 15 minutes of suckling cortisol release was significantly above baseline in 77% of the observed suckling events. Dihydroxyphenylalanine (DOPA) increased in high energy diet cows compared with that of low energy diet cows (2,833 +/- 243 vs 1,294 +/- 243 pg/ml, P<0.01). Norepinephrine (NE) and 3,4-dihydroxyphenylacetic acid (DOPAC) were not influenced by treatment. Plasma NE decreased during the postpartum interval (P<0.005). These data suggest that reduced energy intake may prevent the increase in LH associated with increasing days post partum and alter adrenal function. In addition, spontaneous suckling events elicit a release of cortisol.     

Fertility in heifers and cows after low dose insemination with sex-sorted and non-sorted sperm under field conditions

The present study was performed to test fertility after low dose insemination with sexed and non-sexed sperm in dairy cattle under field conditions in Switzerland. Spermatozoa were stained with Hoechst 33342 and sorted by flow cytometry. A total of 132 heifers and cows were inseminated with 2 x 10(6) X-bearing, frozen-thawed sperm (A) and 91 animals were inseminated with the same dose using non-sorted, frozen-thawed sperm (B). Pregnancy examination by ultrasound was performed twice, 30-40 days (PE1) and 70-90 days (PE2) after insemination. The pregnancy rates after PE1 were 33.3% (9/27) and 59.3% (16/27) in heifers (P=0.05) and 27.6% (29/105) and 28.1% (18/64) in cows (P>0.05) for groups A and B, respectively. Embryonic losses between PE1 and PE2 in heifers were 11.1% (1/9) and 0% (0/16) and in cows 17.2% (5/29) and 5.6% (1/18), the differences between groups A and B not being significant (P>0.05). Calving rates in heifers were 29.6% (8/27) and 57.8% (15/26), whereas in cows 22.1% (23/104) and 23.4% (16/63) gave birth to calves (for both groups P>0.05). The sex ratio was different (P<0.05) between A (85.3%) and B (58.6%). From our results it can be concluded that conception rates of sorted and non-sorted semen are similar using an insemination dose of 2 x 10(6). Fertility may be increased by improving sexing technology and animal management.     

Effects of the presence of a dominant follicle and exogenous oestradiol on the duration of the luteal phase of the bovine oestrous cycle.

The presence of a developing dominant follicle may be a factor in the control of the luteolytic cascade mechanism and the number of follicular waves during the bovine oestrous cycle. In this study, ovaries of all animals were examined once a day by transrectal ultrasonography. It was expected that heifers (n = 18) would have two follicular waves if the second wave occurred later than day 10 after oestrus (Expt 1) and that cows (n = 14) would have three waves if the second wave occurred on or before day 10 (Expt 2). The objective of Expt 1 was to determine if absence of a large follicle late in the luteal phase delays luteal regression in heifers that are expected to have two follicular waves. Nine heifers were injected i.v. with 10 ml charcoal-treated bovine follicular fluid three times a day for 4 days, starting on the day after initiation of the second follicular wave, to delay growth of the second wave dominant follicle. Nine heifers were injected with 0.9% NaCl as controls. The duration of the luteal phase (calculated as the number of days that serum progesterone was > 0.5 ng ml-1) was greater (P < 0.01) in the follicular fluid-treated group compared with the controls (18.7 versus 14.1 days). FSH and follicular growth were suppressed during the period of injection of follicular fluid (P < 0.01 and 0.03, respectively). The objective of Expt 2 was to determine the effect of increased oestradiol on the duration of the luteal phase in cows that were expected to have three follicular waves. Seven cows were injected i.m. three times a day for 4 days with 1 ml oestradiol (100 micrograms ml-1 in corn oil) and seven cows were similarly injected three times a day with 1 ml 0.9% NaCl (control) starting the day after cessation of growth of the second wave dominant follicle. Luteal phase duration was shorter in oestradiol-treated animals than in the controls (14.0 versus 19.0 days; P < 0.04). Serum oestradiol concentrations were higher in the oestradiol-treated group during the period of injection (P < 0.01). In summary, luteolysis was delayed when follicular growth was suppressed with follicular fluid (Expt 1). Exogenous oestradiol administration during the development of uterine oestradiol responsiveness initiated luteolysis earlier compared with control animals (Expt 2).     

Estrus synchronization and pregnancy rates in cyclic and noncyclic beef cows and heifers treated with syncro-mate B or Norgestomet and Alfaprostol

Postpartum lactating cows (N=118) and virgin heifers (N=60) were treated with subcutaneous Norgestomet implants for nine days and received either an intramuscular injection (im) of 5 mg estradiol valerate and 3 mg Norgestomet at the time of implant insertion or an im injection of 5 mg Alfaprostol 24 hr before implant removal. Animals were artificially inseminated 12 hr after detection of estrus. Of the cows and heifers, 78% and 88%, respectively, were in estrus within five days after implant removal (P<0.09). There was no difference between treatments in the proportion of animals in estrus or in the timing of estrus (P<0.85). Estrus was detected in a greater (P<0.05) proportion of animals that were cyclic prior to treatment (88%) than among those that were anestrous prior to treatment (77%). Pregnancy rates after five days were similar between heifers that were cyclic (42%) or anestrous (47%) prior to treatment; however, the five-day pregnancy rate in cows that were anestrous prior to treatment was 38% lower than that in cows that were cyclic prior to treatment (17 vs 55%, P<0.01). Although the treatments synchronized or induced estrus in both cyclic and anestrous animals, marked variability in estrous response and fertility among previously cyclic or anestrous postpartum cows limited the effectiveness of the treatments.     

Estrus synchronization and superovulation with a subcutaneous gestagen implant (Norgestomet, Intervet) in suckler cows and heifers

Four trials were conducted to investigate the suitability of a gestagen implant (Norgestomet, Intervet) for estrus synchronization and superovulation in suckler cows and heifers kept under field conditions in Germany. In trial 1 out of 17 heifers treated 12 responded with one ovulation each. In trial 2 57 suckler cows were treated at the University experimental farm. Of 23 cows treated sooner and 34 cows treated later than 50 days post partum one (4 %) and 16 (47 %) respectively calved at the expected time. Trial 3 was a field trial involving 126 suckler cows and 21 heifers. Of 24 cows treated sooner and 102 cows treated later than 50 days post partum 17 % and 52 % respectively calved at the expected time. Of the 21 heifers only 19 % calved at the expected time. In trial 4 superovulation of 13 cows and 17 heifers resulted in 62 % and 94 % responding with 10.0 +/- 2.5 and 11.1 +/- 3.0 (x +/- SD ) ovulations per animal respectively.     

Influence of a CIDR prior to bull breeding on pregnancy rates and subsequent calving distribution

We determined whether insertion of a CIDR for 7 days prior to the breeding season enhanced pregnancy rates and altered the date of conception in suckled beef cows mated naturally. Suckled beef cows (n=2033) from 15 locations were randomly assigned to one of two treatments: (1) cows received a CIDR 7 days prior to the breeding season for 7 days (CIDR; n=999); (2) cows received no treatment (Control; n=1034). On the first day of the breeding season bulls were introduced to herds at a rate of 15-25 cows per yearling bull or 20-30 cows per mature bull. Pregnancy status and the date of conception were determined via transrectal ultrasonography at 56 and 120 days after initiation of the breeding season. Overall pregnancy rates ranged from 59.3 to 98.9% among the 15 locations. The percentage of cows becoming pregnant during the first 30 days of the breeding season was similar between CIDR (68.2%) and Control (66.7%) cows, and overall pregnancy rates were similar between CIDR (88.9%) and Control (88.6%) cows. The average day of conception after initiation of the breeding season was shorter (P<0.01) for CIDR (20.1+/-0.8 days) compared to Control cows (23.2+/-0.8 days). Of cows conceiving during the breeding season, more (P<0.05) CIDR cows (35.9%) conceived during the first 10 days of the breeding season than Control cows (30.8%). Neither body condition score and nor parity affected pregnancy rates or days to conception, whereas pregnancy rates and days to conception were affected (P<0.01) by location and days postpartum. Days to conception were greater for cows that calved within 40 days (31.6+/-1.2 days) of initiation of the breeding season compared to cows calving between 40 and 50 days (25.3+/-1.2 days) prior to initiation of the breeding season, which were greater than those cows calving between 50-60 days (20.0+/-0.8 days) and 60-70 days (21.3+/-1.0 days) prior to initiation of the breeding season. Cows calving greater than 70 days (17.3+/-1.5 days) from initiation of the breeding season had the shortest interval to conception. We concluded that insertion of a CIDR prior to the breeding season failed to increase overall pregnancy rates, but did influence the average day of conception.     

Effect of restricted suckling on pregnancy rates and calf performance in Brahman cows

Seventy-six Brahman cows and first-calf heifers were assigned to one of two groups: 1) normal suckling (34 cows) or 2) twice-daily suckling (45 minutes of suckling each time; 42 cows). Twice-daily suckling was carried out from 30 days postcalving until weaning (seven months). All animals were maintained under artificial insemination for a four-month breeding period. Mean pregnancy rate was 63.06 +/- 0.06% and was influenced by suckling group (P<0.01) and number of parturitions (P<0.05). The pregnancy rates were 33% higher in twice-daily suckled cows. Forty-four percent of the first calf heifers in the twice-daily suckling group became pregnant compared to 9% in the normal suckling group (P<0.01). Twice-daily suckling improved pregnancy rate without depressing preweaning calf performance.     

The increase in plasma C19Delta5 steroids in subcutaneous abdominal and jugular veins of dairy cattle during pregnancy is unrelated to estrogenic activity

Plasma concentrations of dehydroepiandrosterone (DHEA), 5-androstene-3beta,17beta-diol (AED), and 17beta-estradiol (E2) in dairy cows and heifers and AED binding to uterine cytosolic estrogen receptor (ER) were studied. Plasma samples were collected from the subcutaneous abdominal (SA) and jugular (J) veins of heifers and cows in the non-pregnant state and at 15-45, 90-120, 180-210, and 250-280 days of pregnancy (N = 5-12). Plasma DHEA, AED, and E2 were determined by RIA. DHEA and AED significantly increased (P < 0.001) in heifers and cows throughout pregnancy. The stage of pregnancy significantly (P < 0.001) affected the three steroids in heifers and cows. Plasma DHEA increased throughout pregnancy in both heifers and cows, and in heifers it was significantly greater in SA than in J veins at 90-120 days (P < 0.01). Plasma AED was greater in heifers than in cows in J veins at 90-120 days (P < 0.01) and 180-210 days (P < 0.05), and in SA veins, at 15-45 days (P < 0.01) and 90-120 days (P < 0.05). In heifers, circulating AED showed concentration values significantly greater than those in non-pregnant animals from 90 to 120 days (P < 0.05) and was significantly greater in SA than in J veins at 90-120 days (P < 0.05). In cows, plasma AED was significantly greater than in non-pregnant animals at 250-280 days (P < 0.01). In heifers, plasma E2 was significantly greater in the SA than in the J veins from 180-210 to 250-280 days (P < 0.01). In cows, differences between E2 plasma concentrations in J and SA veins were observed only at 250-280 days of pregnancy. At 250-280 days, in both animal types plasma E2 was significantly greater than in non-pregnant animals (P < 0.001). We suggest that AED originates primarily from the feto-placental unit, while mammary E2 synthesis near term can affect plasma concentrations. Binding data showed that AED is a weak competitor for cytosolic ER (IC50 range: 1.44 x 10(-5) to 3.71 x 10(-5) M). These results suggest that a direct estrogenic activity for AED is unlikely in dairy cattle, and the physiological role of AED needs to be elucidated.     

Uterine blood flow during the first 3 weeks of pregnancy in dairy cows

Transrectal color Doppler sonography was used to compare changes in uterine blood flow between cyclic and early pregnant lactating dairy cows. Examinations were carried out in 53 multiparous lactating cows on days 3, 6, 9, 11, 13, 15, 18 and 21 (day 0=estrus). Fourteen cows were examined during the estrous cycle and thirty-nine cows after insemination with frozen/thawed sperm. Uterine blood flow was reflected by the time-averaged maximum velocity (TAMV) and the pulsatility index (PI) in the uterine artery ipsilateral to the corpus luteum. Twenty-one cows that were not pregnant on day 25 were excluded from the study. There was high inter-individual variability in PI- (CV: 22-62%) and TAMV-values (CV: 22-42%) at all examinations. In cyclic cows, TAMV values increased between days 13 and 18 and PI values decreased between days 15 and 21 (P<0.05). In pregnant cows (n=18), TAMV values increased from days 9 to 11 and then decreased to minimal values by day 18 (P<0.05). The PI values decreased between days 3 and 11 and then increased to maximum levels on day 18 (P<0.05). On day 21, both variables reached (P<0.05) values that did not differ (P>0.05) from those on day 11. The changes in TAMV were correlated with estrogen and progesterone concentrations (r=0.69 and -0.70, respectively; P<0.05) in cycling cows, but not in pregnant cows (P>0.05). The PI values did not correlate with steroid hormone levels (P>0.05). Differences in uterine blood flow between cycling and early pregnant cows were observed only on day 18 (P<0.05). The results show that in pregnant cows changes in uterine blood supply can be detected already in the second week after insemination; these changes do not occur in the second week of the cycle.     

云南省奶牛胚胎移植试验

实验用12头黑白花奶牛（3-9岁）,PGF[2α]作同步发情处理后,周期第10-12天,总剂量36 mg的FSH-P按递减方式每日肌肉注射2次,间隔12小时,共注射4天,在第3天加注PGF[2α]35 mg,进行超数排卵处理。12头供体牛共得胚135枚,可用胚数118枚,为总胚数的87.47%。平均每头得胚11.25枚。胚胎分别移植给20头同步发情处理的黑白花奶牛和20头黄牛受体（有3头为自然发情）,每头移植1枚胚胎。结果,奶牛受孕率为35%（受孕7头,产犊7头）;黄牛受孕率为40%（受孕8头,流产2头,产犊6头）。奶牛和黄牛总受孕率为37.5%。     

Effect of bleeding stress and variable suckling intensity upon serum luteinizing hormone in Brangus heifers

In the first experiment, the effect of the stress of blood collection (via tail vessel puncture) on serum luteinizing hormone (LH) was evaluated in six nonsuckled first calf Brangus heifers. The animals were bled on days 22 and 31 postpartum at 15 minute intervals for a period of two hours. Blood was processed to yield serum and analyzed for LH via radioimmunoassay (RIA). There were no significant differences or fluctuations in serum LH levels between bleeding periods or between cows. Serum LH concentrations in nonsuckled cows were not affected by the stress of blood collection. In the second experiment, 24 first calf Brangus heifers were randomly assigned to one of four treatment groups. Treatment 1 cows were suckled once daily for approximately 30 min starting day 21 postpartum. Treatment 2 cows were suckled twice daily for approximately 30 min each time, starting 21 days postpartum. Treatment 3 cows were suckled once daily for approximately 30 min starting 30 days postpartum. Treatment 4 cows were suckled twice daily for approximately 30 min each time starting 30 days postpartum. Each cow was bled via tail vessel puncture on days one and nine following the start of each treatment. The blood sampling regime was similar to that used in Experiment 1 and consisted of four presuckling samples taken at 15 min intervals, one midsuckling sample (the calf was allowed to suckle for 15 min) and four postsuckling samples taken at 15 min intervals. Blood was collected, processed to yield serum and assayed for LH via RIA. Suckling intensity (SI) was found to have a significant effect on serum LH levels. The once daily suckled cows had higher (P<.01) mean serum LH levels than did the twice daily suckled cows (1.70 +/- .03 and 1.53 +/- .03 ng/ml, respectively). The LH concentrations decreased (P<.01) from the first to last bleeding time (BT). The mean serum LH levels for the presuckling, midsuckling and the first postsuckling samples were higher (P<.05) than the last postsuckling sample. The mean serum LH level for the first time period prior to suckling was higher (P<.05) than the last postsuckling sample. The mean serum LH level for the first time period prior to suckling was higher (P<.05) than the last two periods after suckling (1.73 +/- .08 ng/ml vs 1.51 +/- .06 and 1.41 +/- .06 ng/ml). Bleeding day (BD) and weaning day (WD) did not alter serum LH levels. The interactions found to be significant (P<.01) were SIxBD, SIxWD, BDxWD and BTxSIxBDxWD.     

Pregnancy rate following GnRH + PGF 2alpha treatment of low body condition, anestrous Bos taurus by Bos indicus crossbred cows during the summer months in a tropical environment

Anestrous and lactating Bos taurus by Bos indicus crossbred cows with minimum body condition were studied to determine the efficacy of GnRH+PGF 2alpha combinations for induction of estrus and/or ovulation on pregnancy rate during the months of the year when temperatures are greater. On day 0 (start of treatment), cows were assigned randomly to either treatment or control groups. Treated cows (n = 74) received i.m. 200 microg of GnRH on day 0 and 150 microg of PGF 2alpha 7 days later (day 7). On day 7, treated cows were equally distributed to each of three protocols: (1) Select Synch (n = 25), artificial insemination (AI) 12 h after exhibiting estrus from day 7 (PGF 2alpha injection) until day 12; (2) Ovsynch (n = 24), 200 microg of GnRH at 48 h after PGF 2alpha (day 9) + timed-AI (TAI) 16-20 h later; (3) CO-Synch (n = 25), 200 microg of GnRH + TAI at 48 h after PGF 2alpha (day 9). Control cows (n = 25) received no treatment + AI 12 h after exhibiting estrus from days 0 to 12. Detection of estrus was performed daily during the early morning and evening hours from days 0 to 7 in all the cows, and from days 7 to 12 in the cows treated with Select Synch and in the control group, with the aid of a sterilized bull. Palpation per rectum and transrectal ultrasonography were used on days -30, -20, -10 and 0 to confirm anestrus (absence of CL and no signs of estrus at each evaluation) but with ovarian follicles > or = 10 mm on day 0. Pregnancy rate was 0% for Select Synch, 21% for Ovsynch and 28% for CO-Synch (P < 0.05). In conclusion, the Ovsynch and CO-Synch protocols resulted in greater pregnancy rates compared with the Select Synch protocol in Bos taurus/Bos indicus cows with minimum body condition that were anestrous and lactating during the summer months in a tropical environment.     

Fertility of swamp buffalo following the synchronization of ovulation by the sequential administration of GnRH and PGF2alpha combined with fixed-timed artificial insemination

This study evaluated fertility in swamp buffalo after synchronization of ovulation combined with fixed time artificial insemination. At the start of the study, designated day 0, from a group of 98 female Thai swamp buffalo, 55 buffalo (heifers n° = 20 and cows n° = 35) were selected to be synchronized with GnRH (Day 0) followed by PGF₂alpha (Day 7) and a second treatment with GnRH (Day 9). All buffalo were inseminated at two fixed times 12 h and 24 h after the second injection of GnRH (Ovsynch+TAI group); a second group of 43 buffalo (heifers n° = 19 and cows n° = 24) were not treated and were artificially inseminated (AI) at natural estrus (AI group). Blood samples were taken 22 days after insemination to evaluate progesterone plasma levels. In the Ovsynch+TAI group, overall conception rate (CR; i.e. the number of cows with progesterone >4.0 ng/ml on day 22 after AI divided by the number of animals inseminated), was 38.1% and overall pregnancy rate (PR; i.e. the number of cows that were pregnant at day 50-60 after insemination divided by the number of animals inseminated), was 32.7%. In the AI group overall CR and PR was 34.9%.Within the Ovsynch+TAI group, CR and PR were reduced (P < 0.05) in heifers compared with cows (CR 15.0% vs. 51.4% for heifers and cows, respectively; PR 15.0% vs. 42.9% for heifers and cows, respectively). Within the AI group the efficacy of treatment was similar between heifers and cows (CR and PR 31.6% for heifers and 37.5% for cows).In conclusion, this study indicates that in swamp buffalo it is possible to synchronize ovulation and use timed artificial insemination with the Ovsynch+TAI protocol.     

Fertility response of yearling beef heifers after prebreeding energy manipulation, estrous synchronization and timed artificial insemination

The effect of dietary energy and weight class on the fertility of yearling beef heifers (Angus, Hereford, and Angus x Hereford) was investigated over 2 years. In year 1, heifers (n=58) were classed as heavy (HW; > or =340 kg) or light weight (LW; <340kg) and then assigned to receive either a low (LE; 0.23 kg/day) or high energy (HE; 0.68 kg/day) diet. In year 2, heifers (n=60) were also classified as heavy (> or =335kg) or light weight (<335 kg), but the energy content of the diet was raised so that heifers on the LE and HE were targeted to gain 0.46 and 0.79 kg/day, respectively. Heifers in the four groups, LELW (n=14 and 12), LEHW (n=16 and 17), HELW (n=13 and 15), and HEHW (n=15 and 16) received restricted amounts of concentrate (HE > LE) and free choice hay over 47 or 42 days (year 1 and year 2, respectively). To synchronize estrus, heifers were fed capsules containing MGA (0.5 mg/animal each day) beginning 11 days before the end of the feeding trial (day 0), PGF(2alpha) (25mg i.m.) and estradiol benzoate (Ebeta; 400 microg i.m.) was given on days 8 and 10, respectively. Estrous behavior was observed (days 10 and 11) and all heifers were inseminated on day 11. Following AI, heifers were re-grouped and a bull was introduced (days 27-39) for the second service in both years. Pregnancy diagnosis for the first (days 41-42) and second services (days 69-97) was performed by transrectal ultrasonography. Transrectal ultrasonic observations of ovarian follicle number and size were completed for a subset of heifers (n=5-8) from each experimental group at the end of the feeding trial. The effect of year was not significant for any of the reproductive performance variables measured. The mean ADG was (0.72 +/- 0.04 kg/day) and was greater in LW than HW heifers and in heifers in the HE than LE treatment groups (P <0.05). In heifers receiving the LE diet, ADG was lower in HW than LW heifers (weight x diet; P=0.02; 0.54 +/- 0.04 and 0.62 +/- 0.03 kg/day for HW and LW heifers, respectively). The diameter of the largest follicle was greater in heifers receiving the HE diet (P < 0.05; 11.3 +/- 0.4 mm) than those on the LE diet (10.3 +/- 0.3), and in LW (P <0.05) compared to HW heifers. The HE diet increased the size of the largest follicle in LW but not HW heifers (diet x weight, P <0.05). The percentage of pubertal heifers at the end of the feeding period (59.3%), estrous response (56.4%), conception rate (47.7%), ovulation rate (88.9%), and first service pregnancy rate (36.2%) were not significantly affected by initial weight or diet. There was a tendency for first service pregnancy rates to be greater in LW than HW heifers consuming the LE diet (diet x weight, P <0.1; 54.2 +/- 15 and 30.3 +/- 10% for LELW and LEHW heifers, respectively). Pregnancy rate after two services was greater (P=0.01) in LW (82 +/- 10%) than in HW (64.5 +/- 10%). The LE diet achieved moderate rates of gain and allowed high level of reproductive performance in LW but not HW heifers.     

Effects of dry season nutritional supplementation on growth, onset of puberty and subsequent fertility in Boran and Boran x Friesian heifers in Ethiopia

Shortly after weaning at 8 months of age, 43 Boran and 46 Boran x Friesian crossbred heifers were randomly divided to either receive a supplementary feed containing 16% crude protein during the dry season or to serve as controls. Heifers were examined monthly, and data on body weight, wither height and body condition score were recorded. Ovarian size and structures were determined per rectum and heifers were kept under continuous observation for standing estrus. Blood samples were collected at 10-day intervals every month for determination of plasma progesterone levels. All heifers were exposed to intact bulls for 4 months after they attained 18 months of age. Average daily weight gain to puberty was significantly (P<0.01) higher in heifers given supplementation than in the controls (360 vs 326 g/day). The average daily weight gain in Boran heifers (296 g/day) was significantly (P<0.01) lower than in crossbred (392 g/day) heifers. Mean age at puberty differed significantly (P<0.001) between heifers given supplementation (573 days) and the controls (627 days). Boran heifers attained puberty at a significantly later age (660 vs 540 days; P<0.001) than crossbred heifers. Body weight at puberty did not differ between supplemented and control heifers (226 vs 222 kg); while the difference between Boran and crossbred heifers was significant (216 vs 232 kg; P<0.01). Pelvic size and body condition score were not influenced by supplementation but differed significantly between genotypes. Pregnancy rate after 4 months of breeding was higher in supplemented (79%) than in control (64%) heifers. Boran heifers had a significantly (P<0.001) lower pregnancy rate than crossbred heifers (52 vs 91%). Mean age and body weight at conception were not affected by supplementation but differed significantly (P<0.05) between genotypes. The results indicate that prepubertal supplementary feeding during the dry season increased growth rate, reduced age at puberty and improved fertility in both Boran and Boran x Friesian heifers.