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The ability of fetal calf serum, new-born calf serum and normal steer serum to promote the in vitro development of bovine morulae.

The objective of this study was to compare fetal calf serum, new-born calf serum and normal steer serum as medium supplements in the development of bovine morulae in vitro . Bovine morulae were cultured in Hams F-10 tissue culture medium (HF-10) supplemented with 5% or 10% (v/v) fetal calf serum (FCS), new-born calf serum (NBCS) or normal steer serum (NSS). Embryos were recovered at slaughter from mixed bred donor cows of mixed breeding following estrus synchronization with prostaglandin and superovulation with follicle stimulating hormone. A total of 88 morulae were recovered, washed in HF-10 + 1% Bovine Serum Albumin and randomly assigned to treatments. Embryos were cultured in microdrops of medium under paraffin oil at 37 degrees C in a 5% CO(2) humidified atmosphere. Observations for stage of development were made every 24 hours. In vitro development was analyzed by assigning to each embryo a value of 0-5 based on the most advanced stage reached (0= no development, 5= development to a hatched blastocyst). Analysis of variance of these data revealed a significant treatment effect (P<.001) while no level effect or treatment x level interaction was apparent. Comparison of treatment means by Duncans new mulitple range test showed that NSS was superior to NBCS (P<.05) which was in turn superior to FCS (P<.05) as supplements of HF-10 in promoting the in vitro development of bovine morulae.

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Improved diagnostic accuracy by repetitive ultrasonic pregnancy testing in sheep.

Operator effects and instrument accuracy in using the Scanopreg ultrasonic pregnancy detector in sheep bred at synchronized estrus were studied in three experiments. In the first study, four operators tested the same 101 ewes at 60 and 80 days after breeding. The only significant difference among the four operators was that one operator consistently underestimated pregnancy. Operators did not differ in their diagnoses between days 60 and 80. In the second study, there were no differences between two operators who tested 239 ewes 90 days after breeding. In the third study, one operator tested 318 ewes 60, 70 and 90 days after breeding. The accuracy of diagnosis of pregnancy was at least 90% on each day tested; the corresponding diagnoses of nonpregnancy were 52, 76 and 79% correct. Some ewes that were initially diagnosed as nonpregnant were correctly recognized as pregnant when tested later than day 60. Most of the missed pregnancies were in ewes carrying a single lamb. A second Scanopreg test on day 90 of ewes not diagnosed pregnant on day 60 or 70 identified additional ewes as pregnant. Paired tests (days 70 and 90) recognized 99% of the ewes that eventually lambed.

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Possibility of reducing the luteolytic dose of cloprostenol in cyclic dairy cows.

The administration of cloprostenol by intravulvosubmucous (i.v.s.m.) injection at 1 2 and 1 4 of the dose usually given by intramuscular (i.m.) injection, was tested in dairy cows for luteolysis and estrus synchronization. The i.m. injection was used in ten adult cows at the usual dose of 500 mug/animal. Eleven adult cows and 11 heifers were treated i.v.s.m. with a dose equivalent to 250 mug/animal and 125 mug/animal, respectively. Two injections of cloprostenol were administered 11 days apart to the cows not detected in oestrus after a single injection. Forty-three out of the total 46 animals were detected to be in dioestrus at the time of at least one of the injections, as reflected by the plasma progesterone concentrations at the time of treatments. Three out of the 43 animals injected during dioestrus were refractory to the luteolytic effect of cloprostenol; this appeared to be independent of the dosage and the route of administration (refractory cows were: one adult cow treated i.m. and two treated i.v.s.m. with 125 mug of cloprostenol). The mean time interval from injection to the onset of heat was 82.8 hours with a confidence limit for 95% of probability between 67.9 hours and 92.7 hours. The difference between treatments is not significant. The results suggest that in heifers and adult cows cloprostenol can be given i.v.s.m. route at a reduced dose of 1 4 of the usual 500 mug i.m. dosage without affecting the luteolytic effect of the drug or fertility.

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Embryo transfer from goats seropositive for caprine arthritis-encephalitis virus.

Twelve attempts were made to isolate caprine arthritis-encephalitis virus (CAEV) from the uterine flushings of serologically positive superovulated does mated to serologically positive bucks. Embryos were transferred to eight serologically negative estrus-synchronized recipient does and the recipients were monitored serologically following embryo transfer. Virus isolation was attempted from colostrum and placental tissues from does that kidded following embryo transfers and the surviving kid was monitored serologically until four months of age. The CAEV was not isolated from any of the uterine flushings, colostrum or placental tissues. All recipients and the kid remained seronegative throughout the trial.

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Real-time ultrasonography for determining pregnancy status and viable fetal numbers in ewes.

The reproductive tracts of 26 estrus synchronized, bred ewes were scanned with a portable 5.0 MHz real-time ultrasound unit within 1 to 6 d postbreeding. Intrarectal scanning was performed on alternate days until Days 28 to 30 and twice weekly until Days 50 of gestation. Transabdominal uterine scans were conducted twice weekly from Days 25 to 65 and continued weekly until parturition. A total of 24 ewes (92%) became pregnant. A nonpregnant ewe was recognized 100% of the time by both methods of ultrasonic screening. Correct identification of a gravid ewe as pregnant was 100% from Days 51 to 150 of gestation using transabdominal real-time ultrasonography. There was a significant association (P < 0.005) between the number of lambs born and the number of fetuses observed using transabdominal real-time ultrasonography after Day 25 of gestation. Accurate differentiation of fetal numbers by transabdominal scanning was 100.0% for ewes carrying one lamb and 97.3% for ewes carrying two lambs at Days 51 to 75 of gestation. Fetal attrition was documented in one ewe at Day 49 of gestation. Hydrops allantois was diagnosed in another ewe at 110 d of gestation. A total of 37 lambs were born to 23 ewes in the project flock. No congenital abnormalities were noted in any of the lambs. Transabdominal real-time ultrasonography is a safe, rapid, accurate and practical method for assessing pregnancy status, fetal number and fetal viability in sheep.

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Superovulation, collection and transfer of embryos and demi-embryos from Boran(Bos indicus ) cows and heifers.

Twenty-three Boran(Bos indicus ) cows and heifers were superovulated with pregnant mare serum gonadotropin (PMSG); a total of four embryos and 4.1 +/- 0.3 (mean +/- SEM) ova per ova-producing donor resulted. Follicle stimulating hormone (FSH-P) was then used to superovulate 49 Boran cows for a total of 106 superovulations, of which 63 (59.4%) produced an average of 3.7 +/- 0.4 (mean +/- SEM) embryos. The embryo production was not influenced by either the season or the number of times(one to five) the cows were superovulated. A higher pregnancy rate was obtained when the selection of Boran recipients was based on their plasma-progesterone values (overall 52.5%, single embryos 63.3%, twin demi-embryos 45.8%) than when they were selected by palpation per rectum only (overall 43.8%, single embryos 50%, twin demi-embryos 36.4%). The twinning rate of twin demiembryos was 62.5%, whereas only single calves were born after transfer of two embryos per recipient. No pregnancies were produced following transfer of twin demi-embryos without zonae pellucidae. Transferring single demi-embryos gave a low pregnancy rate (13.3%). Twelve donor Boran cows (21 superovulations) bred with their fathers resulted in a high rate of early embryonic death; additionally, only 20.9% (overall) of the recipients became pregnant. Estrus synchronization of Boran cows with a progesterone releasing intravaginal device (PRID) for a short period (7 d) combined with one injection of prostaglandin (Day 6) produced a larger number of good quality recipients (70.5%) than using double prostaglandin injections (60%).

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Effects of adrenalectomy on serum concentrations of progesterone during the luteal phase of sheep.

An experiment was conducted to test the hypothesis that the adrenal gland influences luteal activity in sheep. Twelve Finnish Landrace x Southdown ewes were either bilaterally adrenalectomized (n = 6) or sham adrenalectomized (n = 6) during the breeding season. At approximately 37 and 47 d after surgery, all ewes received intramuscular injections of cloprostenol to synchronize estrus. Blood samples were taken via jugular venipuncture at 48-h intervals between 1 and 19 d after the last cloprostenol treatment. Serum concentrations of progesterone were determined in each of these samples. Analysis of variance showed that concentrations of progesterone during the luteal phase were lower (P<0.05) in adrenalectomized ewes than in sham-operated controls, but that patterns of progesterone were similar for both groups of sheep. Based on these results, we conclude that the adrenal gland does not appear to be necessary for initiation of luteal regression in ewes.

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Effect of prostaglandin F2alpha dosage and stage of estrous cycle on the estrous response and corpus luteum function in beef heifers.

Ninety-five normal cyclic crossbred beef heifers were used to determine if the proportions of heifers showing estrus, intervals to estrus and corpus luteum (CL) function were influenced by PGF2alpha dosage and (or) the stage of luteal phase when PGF2alpha was administered. Heifers were assigned randomly to treatments in a 4x3 factorial arrangement. Treatments were 5, 10, 25 or 30 mg PGF2alpha injected either in early (5 to 9 d), mid (10 to 14 d) or late (15 to 19 d) stages of the luteal phase. Jugular samples were taken at 0 h and at 8 h-intervals for 48 h and again at 60 h after PGF2alpha treatment for progesterone assay. Heifers were observed for estrus continuously for 120 h PGF2alpha treatment. The proportion of heifers showing estrus was dependent upon (P<0.05) both dosage of PGF2alpha and stage of luteal phase. Heifers given 5 mg of PGF2alpha showed estrus only if treated during the late stage, while those given 10 mg of PGF2alpha showed a progressive increase of heifers in estrus as stage of luteal phase advanced. The proportion of heifers showing estrus after 25 and 30 mg of PGF2alpha increased from 56% for the early stage to 100% for the mid and late stages. Interval to estrus in heifers showing estrus within 120 h after PGF2alpha treatment did not differ (P>0.05) among dosages but tended (P=0.10) to be longer in heifers treated during the mid luteal stage (67 h) than in heifers treated in the two other stages (56 h). A greater proportion of heifers (P<0.05) showed estrus by 60 h after PGF2alpha when treated during the early and late luteal stages (75.5%) than for heifers treated during the mid luteal stage (30.4%). Patterns of progesterone concentrations were influenced (P=0.08) by the three way interaction of dosage, stage and time. In heifers that showed estrus, rate of decline in progesterone tended (P=0.07) to be slower during the mid luteal stage than during the early and late stages. Progesterone did not drop below 1 ng/ml until 32 h in heifers treated during the mid luteal stage; whereas progesterone dropped below 1 ng/ml by 24 h in heifers treated during the early and late stages. These data may be useful in designing more efficient systems for using PGF2alpha or its analogues in estrus synchronization of beef cattle.

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Effect of treatment with estradiol valerate on endocrine changes and ovarian follicle populations in dairy cows.

A linear-array ultrasound instrument was used to monitor the dynamics of follicular cyst formation following estradiol valerate (EV) administration in postpartum dairy cattle. Twelve cyclic cows were given two intramuscular (i.m.) injections of prostaglandin and F(2alpha) (PGF(2alpha)) 12 d apart to synchronize estrus. On Day 16 (Day 0 = day of estrus) six cows received 10 mg of EV in 1 ml sesame oil; the remaining six cows were treated with 1 ml sesame oil. The ovaries of all cows were scanned rectally each morning from Day 9 until 14 or 30 d post treatment. Plasma concentrations of luteinizing hormone (LH) and progesterone (P(4)) were also determined as objective indices of treatment effects. Day 0 to 16 ultrasound pictures of the ovaries of both control and treated cows were characterized by the presence of a corpus luteum (CL; 19 to 38 mm), several small follicles (<5 mm) and a medium-sized follicle (6 to 28 mm). Following treatment in control cows, the CL regressed gradually, and a preovulatory follicle was identifiable by Day 17 to 18, it increased in size and reached a maximum of 28 to 30 mm by Day 20 after ovulation and was identifiable throughout the rest of the cycle. Administration of 0 mg of EV resulted in a rapid reduction in the size of the CL. Growth of a large follicle was observed in all treated animals around Days 16 to 20, but having reached a maximum diameter of 12 to 24 mm it regressed without resulting in ovulation. Subsequent ultrasound pictures of EV-treated cows were characterized by the absence of a new CL and the presence of medium-sized persistent follicles. Estradiol valerate treatment induced early luteolysis (43 +/- 05 h post EV vs 101 +/- 22 h) and an LH surge (41 +/- 11 h vs 125 +/- 17 h).

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Site of action for endotoxin-induced cortisol release in the suppression of preovulatory luteinizing hormone surges.

The study was conducted to identify the mechanisms of endotoxin/cortisol action in the suppression of preovulatory LH surges in heifers infused with Escherichia coli (E. coli ) endotoxin. The hypotheses tested were that 1) endotoxin stimulates the release of progesterone, possibly from the adrenal leading to the LH blockade; 2) cortisol released in response to endotoxin infusion blocks the synthesis of estradiol at the ovarian level, culminating in a failure of the LH surge. Eight Holstein heifers were given two injections of prostaglandin F(2alpha) (PG), 11 d apart, to synchronize estrus. Starting from 25 h after the second injection of PG (PG-2), the uterus of each heifer was infused either with 5 ml of pyrogen-free water (control, n = 3) or with E. coli endotoxin (5 mug/kg of body weight) in 5 ml of pyrogen-free water (treated, n = 5), once every 6 h for 10 treatments. Blood samples were obtained every 15 min for 1 h before infusion and again 2 h after each infusion, then hourly until 1 h before the next infusion. After the tenth infusion, blood was collected daily until estrus. Serum progesterone concentrations remained at baseline values (< 1 ng/ml) in control and treated heifers. The total amount of progesterone measured starting 24 to 84 h after PG-2 injection was not different between control and treated heifers (P 0.05). In the control heifers, serum estradiol concentrations remained basal (< 10 pg/ml) until 4 h before the LH surge. Serum estradiol concentrations increased to 20 +/- 5.6 pg/ml, 4 h before the LH surge in control heifers (LH surge occurred 60 to 66 h after the PG-2 injection). There were no changes in serum estradiol concentrations in treated heifers during the sampling period, and the concentrations remained < 10 pg/ml. The total amount of estradiol measured in control heifers was higher (P < 0.05) than in treated heifers. The results if this study suggest that increases in cortisol concentrations after the infusion of endotoxin might block the synthesis of estradiol at the ovarian level, resulting in the failure of a preovulatory LH surge to occur.

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Age at puberty, fertility and litter size of ewe lambs reared under different photoregimens.

Age at puberty, fertility and litter size of ewe lambs of synthetic sire and dam strains raised under different photoregimens were determined. The lambs were bred during January, May or September at 30 to 32 weeks of age. Irrespective of birth date, the lambs were reared under continuous light from birth to 5 weeks of age. From 5 to 20 weeks of age, they were kept under 16 hours of light dairy (16L:8D; Treatment A), 8 hours of light daily (8L:16D; Treatment B), or a split photoperiod of 8 hours total light daily (7L:9D:1L:7D; Treatment C). Subsequently, all lambs were exposed to 9 hours of light daily until after breeding. Lambs were exposed to rams for two estrous periods after treatment with fluorogestone acetate-impregnated intravaginal sponges and pregnant mares' serum gonadotropin (PMSG) to induce synchronized estrus. Although the age at puberty (174 days) was similar among treatments, the incidence of puberty prior to progestagen sponge treatment was higher (approximately 50%) for lambs reared under Treatments A and C than under Treatment B. Fertility and litter size of lambs were not influenced by the previous photoperiod history or by sexual maturity, i.e., puberal or prepuberal, at the start of the sponge treatment. However, strain, age and weight of lambs at breeding influenced significantly the reproductive outcome.

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A preliminary study of growth, serum prolactin and reproductive performance of beef heifers grazing Acremonium coenophialum -infected tall fescue.

In an experiment with a 2x2 factorial arrangement of treatments, 32 virgin Angus heifers were used to study effects of tall fescue endophyte Acremonium coenophialum and energy supplementation on reproductive performance. Treatments were feeding of endophyte-free (EF) or endophyte-infected (EI) tall fescue without (0%) and with energy supplementation at one percent group mean body weight (1%). Blood samples were collected at 28-day intervals for prolactin (PRL). Starting 21 days before the breeding season, blood was also collected weekly for progesterone (P(4)) and PRL. After 99 days of grazing, heifers received Syncro-Mate-B (SMB) and were then artificially inseminated over a 30-day period. Heifers on EI-0% had a lower (P<0.05) average final body weight. Highest average daily gain was observed in heifers fed EF-1% (0.98 kg/day), whereas, heifers fed EI-0% had lowest average daily gain (0.53 kg/day; P<0.05). Endophyte-infected fescue depressed serum PRL concentrations (171 and 143 ng/ml vs 70 and 85 ng/ml; EF-0% and EF-1% vs EI-0% and EI-1%, respectively; P<0.05). No differences were observed among treatments in luteal activity before or after SMB, in pregnancy rates after a synchronized estrus or in overall pregnancy rates during the 30-day breeding season. Endophyte-infected fescue decreased average daily gain and depressed serum concentrations of prolactin, but had no effect on reproductive performance as measured in this study.

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Successful culmination of pregnancy and live birth following the transfer of frozen-thawed buffalo embryos.

A total of 141 embryos was recovered by nonsurgical flushing of the uterus of 31 superovulated buffalo. A total of 66 good quality embryos (Grade I and Grade II) was frozen using 1.4 M glycerol. Forty-two of the frozen embryos were thawed randomly over a 1-year period, and a total of 39 embryos (Grades I, II or III post thaw) were transferred into an equal number of estrus synchronized recipients. Of 11 confirmed pregnancies, 9 calves were born live.

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A field investigation of the effects of bovine viral diarrhea virus infection around the time of insemination on the reproductive performance of cattle.

During a study of methods of synchronizing estrus in Bos indicus cattle, blood was collected from 169 heifers and 38 cows 2 to 3 days prior to artificial insemination (AI), and then again at Day 51 and Day 210 after AI to determine the incidence of infection with bovine viral diarrhea (BVD) virus. Prior to insemination 53 and 68% of the cows and heifers, respectively, were seronegative to the BVD virus. At Day 51 after AI, 70 and 32% of the seronegative cows and heifers, respectively, had seroconverted; but between Day 51 and Day 210, only 17 and 3% of the seronegative cows and heifers, respectively, had seroconverted. The Day- 51 pregnancy rate of cows which were immune (seropositive) to BVD virus infection at the time of AI was similar to the rate of the cows which became infected around the time of AI. However, the pregnancy rate of the immune heifers (44%, n=54) was significantly (P=0.04) greater than the rate of the heifers which became infected around the time of AI (24%, n=37). It was concluded that infection of susceptible females with BVD virus around the time of AI may significantly lower the pregnancy rate.

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Effect of sometribove administration on LH concentration in dairy cattle.

Sixteen lactating, normally cycling Holstein cows were used to study 1) the effects of sometribove (recombinant methionyl bST) administration on basal LH concentrations and 2) the pituitary response to a GnRH challenge during the breeding period. The cows received a 3-injection regimen of PGF2a for estrus synchronization between 40 and 50 d post partum, and were assigned to a control group or to sometribove treatment (25 mg/d), which began 54 to 59 d post partum. Duration of the bST treatment was 24 d. On Day 10 of the estrous cycle following the third PGF2a injection, blood samples were collected to determine basal concentrations of LH. Immediately following this collection period, a GnRH challenge (100 mug) was used to measure the responsiveness of the pituitary based on the subsequent LH peak. Milk production of sometribove-treated cows increased 7% (2.1 kg/d) above that of the controls. Sometribove did not affect basal or GnRH-induced LH concentrations. Plasma progesterone concentrations during the luteal phase were similar between treatments. Sometribove increased milk yield with no apparent effects on basal or GnRH-induced LH concentrations during the breeding period.

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Effect of porcine reproductive and respiratory syndrome virus (PRRSV) on development of porcine fertilized ova in vitro.

Fertilized pig ova, collected from 14 estrus-synchronized gilts 32 h after presumed ovulation were microinjected or cultured for 72 h in Beltsville Embryo Culture Medium-3 with or without Porcine Reproductive and Respiratory Syndrome Virus (PRRSV). To detect virus in the samples, virus isolation of swine alveolar macrophage, Reverse Transcriptase Polymerase Chain Reaction and Fluorescent Antibody techniques were employed. Microinjection or incubation of embryos with PRRSV did not significantly inhibit development of the porcine embryos in vitro when compared with that of controls (P = 0.75 and P = 0.14, respectively). Although either 10 to 20 TCID50 were microinjected or large concentrations of virus were used for embryo exposure by incubation, PRRSV was not detected in association with the embryos. It is concluded based on the experiments reported here that 4- to 16-cell stage pig embryos are not susceptible to productive infection with PRRSV in vitro.

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Is nutritional anestrus precipitated by subfunctional corpora lutea in beef cows?

Thirty-four multiparous, lactating, cyclic beef cows which calved in moderate body condition were used to determine effects of restricted nutrition on corpus luteum (CL) development and endocrine status. At 78 d postpartum, six cows were assigned to a control (CON) diet (26.0 Mcal ME), fed to increase bodyweight (BW) and body condition score (BCS), and the remaining 28 cows were fed to lose BW and BCS on a restricted (RES) diet (14.0 Mcal ME). Following a 40-d adjustment period on respective diets, estrous cycles were synchronized and cows bled daily for determination of progesterone (P4), luteinizing hormone (LH) and insulin (INS) beginning at the synchronized estrus. Ultrasonography was used to determine the ovulatory follicle and CL development. Control cows were maintained for one estrous cycle and were ovariectomized on day 11 of their second cycle. Ten cows on restricted diet (RES-C) continued to form a functional CL (P4 > 1.5 ng/ml at day 10 of an estrous cycle) through as many as 5 cycles, after which observations were discontinued. Fourteen cows on restricted diet (RES-A) were ovariectomized on day 11 of a cycle when a CL was identified by ultrasonography, but was subfunctional (P4 < 1.5 ng/ml on day 10 of that cycle). Four additional RES-A cows which had subfunctional CL were not ovariectomized but were bled for an additional 25 d. At ovariectomy, CL and ovarian weights were collected. Luteal tissue was prepared for evaluation of P4 synthesis, LH responsiveness in vitro, and for determination of P4 content and total LH receptors. Bodyweight and BCS increased in CON cows; whereas, RES cows lost BW and BCS (P < .05). In the cycle prior to ovariectomy, serum P4 and LH were not different in 18 RES-A cows which developed subfunctional CL in comparison to CON cows. Four RES-A cows not ovariectomized but bled for an additional 25 d neither exhibited estrus, ovulated, nor had P4 concentrations greater than .3 ng/ml. Serum INS was lower in RES-A cows during the cycle prior to ovariectomy than in CON cows (P < .05). During the 11-d period prior to ovariectomy, mean serum P4 and INS were lower in RES-A cows than in CON cows (P < .05); however, serum LH was not different. Furthermore, CL and ovarian weights, P4 content of CL, secretion of P4 by luteal tissue in response to LH in vitro and LH receptor number were not different between CON and RES-A cows. In conclusion, nutritional anestrus may be preceded by the formation of a CL with lower steroidogenic output in vivo. However, luteal tissue, collected from RES-A cows, did not appear to be subfunctional during in vitro incubation when substrate availability and gonadotropin support were equal between diets.

Anestrus↗

Factors affecting retention of early pregnancy in dairy cattle.

Potential factors affecting retention of pregnancy during weeks 5-9 of gestation were studied in dairy cows and heifers (N = 211) on two farms. Cows were examined by ultrasonography for presence of a viable embryo, and sizes of the corpus luteum (CL) and of follicles > or = 5mm were recorded. Blood samples were taken at each examination and assayed for progesterone and estradiol. Overall pregnancy loss was 11.4%. Cows with two CL did not have greater concentrations of progesterone than cows with one CL and they retained fewer pregnancies (P < 0.01; 73% versus 91%). Pregnancy retention was associated positively with concentrations of progesterone and estradiol during week 5 (P < 0.05). Embryos that were lost apparently died before CL regression. Retention of pregnancy declined in cows with high body condition and as age of the cow increased. Pregnancy retention was lower in cows bred to one of four frequently-used service sires (P < 0.05). Days postpartum, milk production, parity, service number, inseminator, synchronization of estrus, diameter of follicles and size of CL did not affect pregnancy retention. In conclusion, retention of pregnancy during placentation varied with concentrations of progesterone and estradiol, age of cow, body condition and service sire.

Abortion, Veterinary↗