Influence of reproductive status on the daily rhythms of oxidative stress markers in <Emphasis Type="Italic">Ovis aries</Emphasis>

Changes in circadian rhythms of dROMs, Oxy-ads and SHp during reproductive stages were studied in Comisana ewes. Twelve ewes were divided in two equal groups. The experimental group consisted of ewes undergoing gestation and lactation following artificial insemination and the control group consisted of non-pregnant ewes. Blood samples were collected every 3 h over a 24 h period, 20 days before insemination, on days 100 and 140 of pregnancy, on days 10, 30 and 200 post-partum and during the dry period. In the control group, blood samples were collected on the same days and with the same procedures as those used for the experimental group. A significant effect of time on all parameters studied was observed in the experimental group. Daily rhythms of the parameters studied were observed in the control group in all experimental conditions, and in the experimental group during pre-pregnancy and dry periods. We conclude that the reproductive status of sheep affects oxidative stress markers in blood and their circadian rhythms.     

Reproductive response of ewes synchronized with different lengths of MGA treatments in intrauterine insemination program

A total of 415 fat tailed ewes were randomly assigned to two groups to assess the effect of duration of melengestrol acetate (MGA) (9 versus 12d) administration on reproductive parameters associated with laparoscopic artificial insemination. At the end of MGA treatment, ewes in each group were subdivided and inseminated with one of two different insemination doses (10×10(7) or 20×10(7) sperm per 0.5 ml insemination dose) of fresh diluted semen. Inseminations were carried out 11-18 h after first detected estrus. Ewes were screened for their return to oestrus from 10 to 21 days post AI and inseminated at their returned oestrus. Pregnancy diagnosis was done from approximately 55 days after insemination in both synchronized and return estrus. For short (9-day) and long (12-day) term MGA treated groups, estrus rates were 62% versus 89% (P<0.0001), respectively. Ewes (n=115) that returned to estrus were inseminated (7-11h after estrus detection) with fresh diluted semen at different doses (20×10(7) or 40×10(7) or 60×10(7) sperm per 0.5 ml insemination dose). Pregnancy rates were 41% and 44% for short term and long term MGA treated ewes, respectively. Pregnancy rate of ewes which returned to oestrus was 53.4%. There was a significant (P<0.05) increase in pregnancy rates (38-52% for 11-16 h; 63% for 17-18 h) when insemination was held at 17-18 h after first detected estrus following MGA treatments. Pregnancy rates were found to be similar in ewes inseminated with 10×10(7) (36%) or 20×10(7) (47%) motile spermatozoa at first AI, and 20×10(7) (44%) or 40×10(7) (59%) or 60×10(7)(48%) at second AI. It was concluded that short term MGA treated ewes were recorded with lower estrus rates but was similar to pregnancy rates with long term MGA treatment. Acceptable pregnancy rates were achieved in MGA induced estrus when insemination is conducted at 17-18 h after estrus onset and with 20×10(7) sperm per insemination dose.     

Determination of gestational age in sheep and goats using transrectal ultrasonographic measurement of placentomes

Three experiments were conducted to determine gestational age in the ewe and doe by measuring placentomes with a B-mode ultrasonograph and a 5 MHz transducer. Transrectal measurements were obtained by placing the female over a bale of hay. In Experiment 1, ewes (n = 12) and does (n = 15) were examined by transrectal ultrasonography every week from breeding to parturition to determine the growth pattern of placentomes during pregnancy. In Experiment 2, placentomes from 132 ewes and 169 does were measured between 30 and 90 d of gestation. A linear regression relationship between fetal age in days and placentome size in mm was calculated and adjusted for does (gestational age = 28.74 + 1.80PL + e, r(2) = 70.34) and for ewes (age = 47.98 + 0.62PL + e, r(2) = 15.59). In Experiment 3, the placentomes of 63 does were measured to validate this relationship by using linear regression. Gestational age was determined correctly in 66% of the does, with a range of +/- 7 d and in 96% with a margin of +/- 14 d. In conclusion, transrectal ultrasonography allowed for the measurement of placentome size, which increased rapidly during the first 70 to 90 d of gestation in ewes and does. In ewes, however, there was a poor correlation of placentome size with gestational age, while in goats, measurement of placentomes could be used along with pregnancy diagnosis by transrectal ultrasonography as an indication of gestation age.     

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.     

Effect of reproductive disorders previous to conception on pregnancy attrition in dairy cows

This study was undertaken to determine whether reproductive disorders previous to conception influence pregnancy attrition in dairy cows. Reproductive disorders were registered and pregnancy diagnoses were performed as a part of a reproductive health program at 9 commercial dairy herds in northeastern Spain. Data from 3022 diagnosed pregnant lactating cows were used. Pregnancy diagnosis by palpation per rectum was performed from 33 to 70 d post insemination. Pregnancy attrition was registered when the pregnancy diagnosis was negative at the second palpation carried out between 120 and 150 d following insemination. Data analysis was performed by multiple logistic regression methods. Pregnancy attrition rates were 2.6 and 1.8 times higher in cows with previous pyometra and retained placenta, respectively, than in cows without these disorders. No effect of endometritis, ovarian cysts and repeat breeding was shown on pregnancy attrition. Our results suggest that additional efforts to reduce the risk of retained placenta and pyometra should decrease the incidence of pregnancy attrition in dairy cows.     

Effect of eCG on the pregnancy rate of ewes transcervically inseminated with frozen-thawed semen outside the breeding season

An experiment was conducted to determine whether factors affecting pregnancy rate out-of-season are associated more with transcervical artificial insemination (T-AI) procedures or with the reproductive state of the ewe. Twenty Finncross ewes were treated with progesterone sponges, and at sponge removal (0 h) 10 ewes were treated with eCG. Blood samples were collected for LH and progesterone analyses, and follicular development was monitored using ultrasonography. Ewes were inseminated from 48 to 52 h with 200 million motile frozen-thawed spermatozoa. The incidence of estrus, LH surges and ovulation was greater (P < 0.01) and intervals to these responses were shorter (P < 0.01) in the eCG-treated ewes. The number of follicles > 5 mm was higher (P < 0.05) in eCG-treated than control ewes. Progesterone concentrations increased and remained elevated through Day 19 in 7 eCG-treated and in 1 control ewe, and these ewes were pregnant based upon ultrasonographic examination. The results demonstrate that the T-AI technique using frozen-thawed semen produces a relatively high (70%) pregnancy rate out-of-season. The pregnancy rate was found to reflect primarily the reproductive condition of the ewe.     

Short-duration insemination with frozen semen increases fertility rate in nulliparous dairy goats

Standard artificial insemination (AI) using a speculum in dairy goats does not result in acceptable fertility rates in nulliparous does. An explanation might be the difficulties to pass the cervical canal in nulliparous females with the insemination gun, increasing the time needed for semen deposition. Nulliparous Alpine dairy goats were used to evaluate whether time interval from insertion to withdrawal of the speculum is a factor influencing pregnancy rates to first AI with frozenthawed semen. Oestrus was synchronized using fluorogestone acetate intravaginal sponges (FGA, 40 mg) for 11 days, associated with 50 mg i.m. of cloprostenol and 250 IU i.m. eCG 48 ± 2 h before sponge removal. In the first experiment (n = 52; 3 herds), the average duration of the AI procedure was 42 ± 10 s, with a median of 39 s. AI performed in less than 39 s resulted in higher pregnancy rates (75%, n = 28) than AI lasting for more than 39 s (46%, n = 24). In the second experiment, does (n = 325; 5 herds) were randomly assigned into two treatment groups according to a short (20 s) or long (60 s) AI procedure. We showed that the duration of AI affected fertility after a first insemination, and that pregnancy rate was significantly improved using a short-duration AI (61.2%; n = 169) compared with a long-duration AI (44.2%; n = 156). We have previously shown in the ewe that genital stimulation during AI enhanced uterine motility. Other authors reported a negative correlation between increased uterine motility at the time of AI and fertility rates in small ruminants. The results of this study suggest that rapid semen deposition may limit the reflex activation of uterine contractions provoked by the speculum and the movement of the insemination gun, and thus ameliorates reproductive performance to first AI in nulliparous goats.     

Progesterone in bovine milk fat

The progesterone (P(4)) concentration (ng/5 mul) in the fat portion of cow's milk was measured on days 0, 12, 20, 24, and 30 after insemination in an effort to assess the ability of this analysis to judge the reproductive status of dairy cows. Pregnancy was determined by rectal palpation at approximately 40 days after insemination. The mean P(4) concentration (ng/5 mul) +/- SD in the pregnant cows (n=17) was 0.14+/-.07, 1.31+/-.36, 1.41+/-.52, 1.22+/-.21, and 1.30+/-.43 on post-insemination days 0, 12, 20, 24, and 30 respectively. At these same intervals the P(4) concentration in the non-pregnant (NP) cows (n=18) was 0.14+/-.06, 1.26+/-.37, 0.56+/-.52, 0.57+/-.42, and 1.08+/-.59. On days 20 and 24 after insemination, mean P(4) levels were significantly (P<.001) elevated in the pregnant cows. It was concluded that an accurate assessment of the reproductive status of a cow, milk samples from days 0, 20, and 24 post-insemination would have to be analyzed for milkfat P(4) concentrations. In order to determine the percentage of cows inseminated out of the periestrual period, milkfat P(4) concentrations were ascertained in milk collected from 165 cows at the time of insemination. Cows that conceived (n=38) had a mean milkfat P(4) concentration at the time of insemination of 0.16+/-.09. The upper limit for P(4) in the milkfat at the time of insemination in cows that conceived was calculated to be 0.43 ng/5 mul milkfat. Subsequently, it was found that of the 14 cows that had P(4) levels above this upper limit at time of insemination, nine were inseminated at a period other than in the periestrual period and five were inseminated while already pregnant.     

Observed and expected combined effects of clinical mastitis and low body condition on pregnancy loss in dairy cows

The objective was to compare the observed and expected combined effects of clinical mastitis before timed artificial insemination (TAI) and low body condition at 70 d postpartum (dpp) on pregnancy loss in dairy cows. Cows were examined for pregnancy by ultrasonographic examination 28-32 d after TAI; the presence of an embryo with a heartbeat was the criterion used to determine pregnancy. Cows diagnosed pregnant were re-examined by transrectal palpation of the uterus and its contents 28 d later to confirm pregnancy status and to identify pregnancy loss. Eighty-eight (17%) of 512 cows were diagnosed with pregnancy loss. Cows affected with clinical mastitis before insemination and a body condition score (BCS) ≤ 2.75 at 70 dpp were 2.03 times more likely to experience pregnancy loss, compared to cows without clinical mastitis and with a BCS > 2.75 (RR = 2.03; 95% CI = 1.15, 3.60; P = 0.01). This observed combined effect for pregnancy loss (RR = 2.03) was higher than the expected combined effect based on adding (RR = 1.39) or multiplying (RR = 1.42) absolute independent excesses due to clinical mastitis or low body condition.     

Evaluation of false transrectal ultrasonographic pregnancy diagnoses in sheep by measuring the plasma level of pregnancy-associated glycoproteins

The present study was undertaken to investigate to what extent pregnancy diagnoses made by transrectal ultrasonography could be confirmed by measurements of plasma concentration of ovine pregnancy-associated glycoproteins (ovPAG). A total of 424 Awassi x Merino ewes were synchronized for estrus and examined by transrectal ultrasonography. In Experiment 1, the ewes (n = 156) were repeatedly scanned in a standing position on d 29, 36 and 50 of gestation. Similarly, the ewes (n = 268) in Experiment 2 were scanned on d 24, 29 and 34 of gestation, but these ewes were fasted for 12 h prior to the examination and the abdominal wall of each animal was lifted up by the hands of the assistant during the scanning. Blood samples were withdrawn after each transrectal ultrasonographic examination in both experiments. Ovine PAG concentrations were measured in plasma by a heterologous radioimmunoassay and the cut-off value for pregnancy was > or = 1 ng.mL-1. Based on the lambing performance, in Experiment 1, altogether 47 false negative and 38 false positive diagnoses were made by transrectal ultrasonography in 24 and 33 ewes, respectively between d 29 and 50 of gestation. In Experiment 2, altogether 8 false negative and 13 false positive diagnoses both were made in 7 ewes between d 24 and 34 of gestation. In both experiments, all ewes with false negative diagnoses had ovPAG concentrations higher than the threshold level for pregnancy diagnosis and all ewes with false positive diagnoses had ovPAG concentrations lower than the threshold of pregnancy. Furthermore, by the PAG-RIA test all lambed or aborted ewes (n = 63) were correctly diagnosed as pregnant and with three exceptions, all non-lambed ewes (n = 361) were correctly diagnosed as non-pregnant during the examined periods of both experiments.     

Transuterine sperm transport is not affected by bilateral asymmetry of the reproductive system in dairy cows

Effects of right versus left side activities of the reproductive organs on sperm transport after deep cornual insemination were evaluated in 1686 Friesian cows in their first lactational period. Only single ovulating animals were used. At insemination, semen was deposited deep into the uterine horn ipsilateral or contralateral to the preovulatory follicle. Pregnancy rates were used as measurement of the success of sperm transport. The reproductive history of each cow included that of the side of the previous gestation. A higher activity of the right versus left side was registered. Following calving and after uterine involution, activities of the right versus left side reproductive organs remained constant independently of the side of previous pregnancy, and did not affect transuterine sperm transport. The data indicate that after uterine involution sperm transport is not affected by bilateral asymmetry of the reproductive system in cattle.     

A large-scale program in laparoscopic intrauterine insemination with frozen-thawed semen in Australian Merino sheep in Argentine Patagonia

This report shows the results of a large-scale laparoscopic intrauterine insemination program on a flock of Australian Merino sheep in Argentine Patagonia. The study was carried out on a total of 1824 ewes (3-to-7-yr-old) and 480 ewe hoggets (19-20 months old) on 2 farms in the southeastern region of Santa Cruz Province, in April and May 1996. The animals, divided into 15 groups, were synchronized with vaginal sponges containing 60 mg medroxyprogesterone acetate for 14 d and injected with 200 IU PMSG upon sponge removal. Estrus was screened every 12 h by means of vasectomized marker rams. The animals were inseminated laparoscopically by the intrauterine route using 2 schemes: 1) at a fixed time (12 h) after estrus detection, or 2) at a fixed time (60 h) after sponge removal irrespective of estrus. Pregnancy was determined at 30 d by transrectal ultrasound imaging. The results showed that 1) the onset of estrus occurs most often between 24 and 48 h after sponge removal, 2) ewe hoggets undergo estrus significantly earlier than sexually mature ewes, 3) in those animals showing estrus, there appears to be no relationship between fertility (as assessed by pregnancy outcome) and time of estrus, 4) there is a significant association between the percentage of estrus occurrence and pregnancy rate, 5) fertility is significantly higher in ewes than in hoggets, 6) for practical purposes insemination at a fixed time after the onset of estrus has no advantage over that of to insemination at a fixed time after sponge removal. It is concluded that large-scale laparoscopic intrauterine insemination can be successfully applied in Australian Merino ewes and ewe hoggets in low-productivity areas such as that of Argentine Patagonia and that estrus detection is unnecessary when insemination is performed at 60 h after sponge removal.     

Embryonic loss from 30 to 60 days post breeding and the effect of palpation per rectum on pregnancy

This study was conducted over a 12-mo period to determine the rate of bovine embryo death between 30 and 60 d of gestation. In addition, palpation per rectum as a means of pregnancy detection was evaluated as a possible cause of embryo death. Estrus was synchronized in Holstein heifers (n = 1358), weighing > or = 385 kg, with a single intramuscular injection of 25 mg prostaglandin F(2alpha). Estrus was primarily detected by the absence of paint marks on the tailhead. The heifers were artificially inseminated with semen from 5 Holstein sires. Blood was collected between 30 and 45 d after breeding, and sera were evaluated for the presence of bovine pregnancy-specific protein B (bPSPB) by RIA to determine pregnancy. Palpation for fetal membrane slip was conducted by an experienced technician in approximately one-half of the inseminated heifers. To determine embryonic survival, a second blood sample was collected at approximately 60 d from 862 heifers that were determined to be pregnant at the first blood sampling. Embryonic loss averaged 5.3% during the interval between the initial detection of pregnancy at 30 to 45 d and the subsequent detection of pregnancy at 60 d of gestation. Embryo loss in heifers that were palpated was 6.5% compared with that of 4.3% in the control heifers (X(2): P = 0.145). These findings establish that there was substantial loss of embryos between 30 and 60 d post breeding but that embryo loss was not affected by palpation per rectum.     

Factors affecting pregnancy loss from gestation Day 38 to 90 in lactating dairy cows from a single herd

The present study was designed to establish whether factors such as previous estrus synchronization, corpus luteum and embryo number at the time of pregnancy diagnosis, changes in body condition score, milk production, clinical disease (mastitis or lameness) and the inseminating bull affect pregnancy loss from 38 to 90 days of gestation. We derived data from 601 pregnant lactating dairy cows from a single herd. Pregnancy diagnosis was performed by ultrasonography between Day 38 and 44 following insemination. We also recorded corpus luteum and embryo number at this time. Pregnancy loss was defined as a negative pregnancy diagnosis on the second palpation per rectum undertaken between 90 and 96 days after insemination. Data were analyzed using multiple logistic regression methods. Cows that had an additional corpus luteum were eight times less likely to miscarry. The risk of pregnancy loss was 3.1 times higher in cows bearing twins. A one unit reduction in body condition score from previous partum to 30 days postpartum resulted in a 2.4-fold increase in pregnancy loss. We noted a higher incidence of pregnancy loss in cows inseminated using semen from one of the six bulls used. This particular bull led to a 3.4-fold increase in the rate of pregnancy loss. Logistic regression analysis showed no significant effects of previous estrus synchronization, milk production, clinical disease, body condition at previous partum or at pregnancy diagnosis, or body condition change between previous partum and pregnancy diagnosis. Our findings indicate a positive relationship between the presence of an additional corpus luteum and the maintenance of gestation. Risk factors for pregnancy loss were twin pregnancy, reduced body condition after previous parturition and the inseminating bull.     

Increase of pregnancy rate in dairy cattle after preovulatory follicle palpation and deep cornual insemination

A total of 334 first-service lactating cows in natural estrus were used in the study. Semen was deposited into the uterine body of 174 cows and deep into the uterine horn ipsilateral to the side of impending ovulation of 160 cows. In both groups, insemination was performed within the interval of 50 to 100 d postpartum at 8 to 15 h after estrus detection and after preovulatory follicle palpation. Pregnancy rates were determined by palpation per rectum 50 d post insemination. The pregnancy rate was higher (P < 0.05) for deep uterine horn insemination (70.62%) than for uterine body insemination (60.34%).     

Pregnancy associated glycoproteins (PAG) in postpartum cows, ewes, goats and their offspring

Determination of plasma concentrations of pregnancy associated glycoproteins (PAG) has been used for early pregnancy diagnosis in cows. However, this is complicated by the presence of PAG in plasma for an extended period postpartum. The main objective of the present study was to investigate the postpartum elimination rates of pregnancy associated glycoproteins (PAG) in sheep, goats and cows in order to gain background information applicable to the use of PAG for pregnancy diagnosis in domestic ruminants. A second objective was to investigate whether PAG are transferred to the foetus and newborn, by measuring plasma PAG concentrations in calves, lambs and goat kids before and after colostrum feeding. PAG in the blood at parturition were eliminated by a first order process in the cows and ewes, while a two-step log-linear decline occurred in the goats. Estimated postpartum half-life of plasma PAG in the cows and ewes was 9 and 4.5 days, respectively. In the goats, half-lives were 3.6 and 7.5 days in the initial fast and terminal slow phase. Basal levels were reached 80-90 days postpartum in cows. Plasma PAG concentration can be used for pregnancy diagnosis from day 28 after AI, provided that the time interval from calving to AI is >60 days. Using a heterologous antibody RIA, we found 4 ng/mL to be the appropriate cut-off. Due to the presence of PAG residues from the previous gestation, the interval from AI to pregnancy diagnosis should increase by approximately 0.5 days beyond 28 days for each day of AI closer to calving than 60. Measurements in newborn ruminants suggested that PAG enter the foetal blood in utero and that colostral PAG are transferred to the newborn. Following the peak plasma concentration observed 1 day after birth in most of the animals, PAG were rapidly eliminated in a log-linear fashion.     

Reproductive capacity of Merino ewes fed a high-salt diet

An option to increase the productivity of saline land is to graze sheep on salt-tolerant plants, which, during the summer/autumn period, can contain 20% to 25% of their dry matter as salt. This study assessed the impact of coping with high dietary salt loads on the reproductive performance of grazing ewes. From the time of artificial insemination until parturition, 2-year-old maiden Merino ewes were fed either a high-salt diet (NaCl 13% of dry matter) or control diet (NaCl 0.5% of dry matter). Pregnancy rates, lamb birth weights, milk composition and the plasma concentrations of hormones related to salt and water balance, and energy metabolism were measured. Leptin and insulin concentrations were lower (1.4 ± 0.09 v. 1.5 ± 0.12 ng/ml; (P < 0.05) and 7.2 ± 0.55 v. 8.2 ± 0.83 ng/ml; P < 0.02) in response to high-salt ingestion as was aldosterone concentration (27 ± 2.7 v. 49 ± 5.4 pg/ml; P < 0.05), presumably to achieve salt and water homeostasis. Arginine vasopressin concentration was not significantly affected by the diets, but plasma concentration of T3 differed during gestation (P < 0.02), resulting in lower concentrations in the high-salt group in the first third of gestation (1.2 ± 0.18 v. 1.3 ± 0.14 pmol/ml) and higher concentrations in the final third of gestation (0.8 ± 0.16 v. 0.6 ± 0.06 pmol/ml). T4 concentration was lower in ewes ingesting high salt for the first two-thirds of pregnancy (162 ± 8.6 v. 212 ± 13 ng/ml; P < 0.001). No substantial effects of high salt ingestion on pregnancy rates, lamb birth weights or milk composition were detected.     

Influence of management factors on pregnancy attrition in dairy cattle

The present study was conducted to determine the influence of management factors on pregnancy attrition in dairy cattle. Data from 3162 diagnosed pregnancies in parous cows and 1050 in heifers at 9 commercial dairy herds in northeastern Spain were used. Pregnancy diagnosis by palpation per rectum was performed from 30 to 70 d post insemination. Pregnancy attrition was registered when pregnancy diagnosis resulted negative in a second palpation carried out between 120 and 150 d following insemination. Overall proportion of pregnancy losses was 7.9% (9.6% in parous cows and 2.8% in heifers). Data analysis was performed by multiple logistic regression methods. For all animals, effect of time of pregnancy diagnosis was shown (Odds ratio = 0.97 for 1 d increase; P = 0.0042). Conceptus loss in heifers was lower than in parous cows (Odds ratio = 0.28; P = 0.0001), and a higher proportion of pregnancy attritions was detected in animals inseminated in spring, summer and winter, compared to those inseminated in autumn (P < 0.04). Herd effect on pregnancy attrition was also significant. Similar results were observed in the subanalysis for parous cows and, furthermore, no effect of lactation number and of interval from previous calving to pregnancy was shown in this group. In heifers, no effect of time of pregnancy diagnosis and of insemination season on pregnancy attrition was shown, and only a herd effect was observed. Our data suggest that the influence of parity status (heifer vs cow) could affect the proportion of pregnancy attrition rather than early diagnosis, and, in pregnant cows, adaptation to seasonal changes associated with temperature decreases seem more efficient.     

Developmental changes in polyamine levels and synthesis in the ovine conceptus

Polyamines (putrescine, spermidine, and spermine) are essential for placental growth and angiogenesis. However, little is known about changes in polyamine synthesis associated with development of the ovine conceptus (embryo/fetus and associated placental membranes). We hypothesized that rates of placental polyamine synthesis were maximal during the rapid placental growth that occurs in the first half of pregnancy. This hypothesis was tested using ewes between Days 30 and 140 of gestation. Columbia cross-bred ewes were hysterectomized on Days 30, 40, 60, 80, 100, 120, or 140 of gestation (Day 0 = mating; n = 4 ewes/day) to obtain placentomes, intercotyledonary placenta, intercaruncular endometrium, and allantoic as well as amniotic fluids. The tissues were analyzed for ornithine decarboxylase (ODC) and arginase activities; arginine, ornithine, and polyamine concentrations; and polyamine synthesis using radiochemical and chromatographic methods. Maximal ODC and arginase activities and the highest rates of polyamine synthesis were observed in all tissues on Day 40 of gestation. Concentrations of ornithine and polyamines in placentomes and intercaruncular endometrium also peaked on Day 40 of gestation. In ovine allantoic and amniotic fluids, polyamines were most abundant during early (Days 40-60) and late (Days 100-140) gestation, respectively. Amniotic fluid spermine increased progressively with advancing gestation. Results of the present study indicate metabolic coordination among the several integrated pathways that support high rates of polyamine synthesis in the placenta and endometrium during early pregnancy. Our findings may have important implications for both intrauterine growth retardation and fetal origins of diseases in adults.     

High fertility using artificial insemination during deep anoestrus after induction and synchronisation of ovulatory activity by the "male effect" in lactating goats subjected to treatment with artificial long days and progestagens

The response to the male effect was studied in two Saanen and two Alpine flocks over 5 consecutive years. Adult male and female goats were exposed to artificial long days (16h light and 8h darkness, 16L:8D) in open barns for approximately 3 months (between December 1 and April 15) followed by a natural photoperiod. Goats were treated for 11 days with fluorogestone acetate (FGA) or progesterone (CIDR) immediately before joining. Bucks carrying marking harnesses with adapted aprons joined females 49-63 days after the end of the long-day treatment (between April 30 and June 5) and were left with them for 5 days. In experiment 1 (n=142), FGA- and CIDR-treated goats were inseminated at a time based on the detection of oestrus. Two insemination groups were distinguished by the occurrence of marking over a 48-h period. Earlier (group 1) and later (group 2) buck-marked goats received one single insemination 12-24h or 0-12h after marking, respectively. Unmarked goats were inseminated along with group 2. In experiment 2 (n=344), FGA-treated goats were inseminated 52 and 70 h (52 h:70 h group) or 52 and 75 h (52 h:75 h group) after joining. In experiment 3 (n=285), FGA-treated goats were inseminated 52 h (1-AI group) or 52 and 75 h (2-AI group) after joining. In all experiments, an external control group given the "classical" insemination program was analysed. Over the 5-year period, 92% of the goats exhibited an LH surge during days 1-4 after joining and 98% of them ovulated. Eighty-seven percent of the LH surges detected in milk occurred during the 33-57 h interval after joining, indicating that ovulation took place around 45-69 h. In experiment 1, 96% of the goats were marked 22-70 h after joining. Kidding rate (KR; 78%) was similar between insemination groups and between FGA- and CIDR-treated goats (p>0.05). Most of the goats (95%) were inseminated during the interval between 15h before and up to 4h after ovulation. KR was not affected by the time between detection of marking and insemination or between insemination and ovulation (p>0.05). In experiment 2, KR (75%) was similar in both insemination groups (p>0.05). In experiment 3, KR was higher (p<0.05) in the 1-AI (71%) than the 2-AI group (57%). In all experiments, KR of the control group (68-73%) was similar to that achieved in goats induced to ovulate by the male effect. Prolificity (2.1+/-0.7) was not affected by any of the factors examined (p>0.05). In conclusion, high fertility can be achieved during anoestrus when 1 or 2 inseminations are performed over a 24h period, determined by oestrus or by the introduction of the buck, if light-treated goats receive 11-day FGA or CIDR treatment and are then induced to ovulate by the male effect.