Intermittent suckling enables estrus and pregnancy during lactation in sows: effects of stage of lactation and lactation during early pregnancy

Previously we demonstrated that pre-ovulatory LH and post-ovulatory progesterone (P4) concentrations in plasma were low and embryo development was retarded when sows were induced to ovulate during lactation by submitting them to intermittent suckling (IS). The present study investigated whether this was due to: (1) stage of lactation when IS was initiated, and (2) continuation of IS post-ovulation. Multiparous Topigs40 sows were studied under three conditions: conventional weaning at Day 21 of lactation (C21; n = 30), intermittent suckling from Day 14 of lactation (IS14; n = 32), and intermittent suckling from Day 21 of lactation (IS21; n = 33). Sows were separated from piglets for 12 h daily during IS. IS sows were either weaned at ovulation or 20 d following ovulation. One-third (21/63) of the IS21 and C21 sows had already ovulated or had large pre-ovulatory follicles at Day 21 and were excluded from further study. Initiation of IS at Day 14 instead of Day 21 of lactation tended to reduce P4 at 7 d post-ovulation (P = 0.07), did not affect pregnancy rate, and tended to reduce embryo survival (P = 0.06). Continuation of IS during pregnancy resulted in lower P4 at 7 and 12 d post-ovulation, tended to reduce embryo weight and pregnancy rate (P < 0.10), whereas embryo survival was not affected. This study presents data for a population of sows in which follicle growth and ovulation are easily triggered under suckling conditions. Further, when these sows are bred during lactation, initiation of IS at 21 rather than 14 d of lactation with weaning at ovulation yields the most desirable reproductive performance.     

Impact of buserelin acetate or hCG administration on day 12 post-ovulation on subsequent luteal profile and conception rate in buffalo (Bubalus bubalis)

The present study investigated the impact of gonadotropic hormone administration on day 12 post-ovulation on subsequent luteal profile and conception rate in buffaloes. All the buffaloes (n = 48) were estrus synchronized by a synthetic analogue of prostaglandin F_2α (PGF_2α), administered 11 days apart, followed by insemination during mid to late estrus. To examine the effect of mid-luteal phase hormonal treatment, buffaloes were randomly divided into control (normal saline, n = 14), d12-BA (buserelin acetate, 20 μg, n = 17) and d12-hCG (hCG, 3000 IU, n = 17) groups. Ovaries were scanned on the day of induced estrus to measure the preovulatory follicle (POF) diameter and on days 5, 12, 16 and 21 post-ovulation to examine the alterations in corpus luteum (CL) diameter. On the day of each sonography, blood samples were collected for the estimation of plasma progesterone. In treatment groups, luteal profile (CL diameter and plasma progesterone) on day 16–21 post-ovulation was better (P < 0.05) as well as first service conception rate was higher (52.9% in each treatment group vs. 28.6%, P > 0.05) compared to controls. All the pregnant buffaloes exhibited higher (P < 0.05) plasma progesterone on various post-ovulation days than their respective non-pregnant counterparts. Treatment-induced accessory corpus luteum (ACL) formation was observed in 58.8 per cent and 70.6 per cent buffaloes of d12-BA and d12-hCG group, respectively, that also had higher (P < 0.05) plasma progesterone compared to controls. Compared to the spontaneous CL, the diameter of ACL was less (P < 0.05) in the treatment groups. In conclusion, buserelin acetate and hCG administration on day 12 post-ovulation leads to accessory CL formation, improves luteal profile and consequently increases conception rate in buffaloes.     

Relationships of age to uterine function and reproductive efficiency in mares

The uterine function and reproductive efficiency of 31 nonlactating pony mares were compared for two age groups: young (5 to 7 years, n=9) and old (>/=15 years, n=22). For pregnant mares, differences between age groups were not significant for the diameter of the largest follicle, cross-sectional area of the corpus luteum, growth profile of the embryonic vesicle or embryo mobility characteristics. Uterine contractility scores were lower (P<0.05), day of fixation of the embryonic vesicle was later (P<0.05), and uterine tone tended (P<0.10) to be lower in the old than the young mares. Endometrial biopsies in old mares had more (P<0.05) inflammatory cell infiltrations, more (P<0.05) fibrotic changes, and less dense (P<0.05) endometrial glands than in young mares. Ultrasonically detected intrauterine fluid collections were more extensive (P<0.05) in the old than the young mares. The pregnancy rate on Day 12 (Day 0=ovulation) was lower (P<0.05) and embryo-loss rate (Days 12 to 39) was greater (P<0.05) in old (32 and 62%, respectively) than in young (100 and 11%, respectively) mares. The results confirmed previous reports that old age was associated with increased endometrial inflammation, reduced pregnancy rate and increased embryo-loss rate. The results also indicated that uterine contractility and uterine tone were reduced and the fixation of the embryonic vesicle occurred later in old than in young mares.     

Factors affecting pregnancy loss for single and twin pregnancies in a high-producing dairy herd

Our objective was to determine the magnitude of, and factors affecting, pregnancy loss for lactating Holstein cows on a commercial dairy farm when diagnosed with twin (n = 98) or single (n = 518) pregnancies using transrectal ultrasonography. Pregnancy losses were assessed with records of non-viable embryos at first pregnancy examination and embryo losses between the first (25-40 d after AI) and second (48 and 82 d after AI) post-breeding pregnancy examinations. Among cows diagnosed with single pregnancies, 3.7% were diagnosed with a non-viable embryo at first pregnancy examination, and 4.6% of those diagnosed with a viable embryo underwent pregnancy loss by the second examination. A total of 11.2% of cows diagnosed with twins experienced a single embryo reduction, whereas 13.3% lost both embryos. Overall, the total proportion of cows experiencing pregnancy loss or experiencing embryo reduction was greater for cows diagnosed with twin than single pregnancies (odds ratio; OR = 3.6), resulting in an embryo survival rate of 91.9% for cows diagnosed with single compared to 75.5% for cows diagnosed with twin pregnancies. Season of breeding and milk production were associated with pregnancy loss for single pregnancies, whereas CL number was associated negatively with embryo reduction and pregnancy loss for twin pregnancies. The risk of twinning and double ovulation among pregnant cows increased with days in milk (DIM), and the risk of double ovulation was greater for cows diagnosed with ovarian cysts and lacking a CL at initiation of an Ovsynch protocol. We concluded that in this herd, embryo reduction and pregnancy loss during early gestation was greater for lactating Holstein cows diagnosed with twin compared to single pregnancies. In addition, cows diagnosed with ovarian cysts and lacking a CL had an increased risk for double ovulation.     

Embryo loss in cattle between Days 7 and 16 of pregnancy

Embryo loss between embryonic Days 7 and 16 (Day 0 = day of IVF) in nonlactating cattle, Bos taurus, was analyzed using transfer of 2449 (in groups of 3 to 30) in vitro-produced (IVP) blastocysts. In 152 transfers, pregnancy losses attributable solely to recipient failings amounted to between 6% (beef heifers) and 16% (parous dairy cows), of which 3% were caused by uterine infections. Neither season, year, nor the age of the embryos on retrieval affected pregnancy rates. The latter observation indicated that the reason that a recipient failed to retain embryos was already present at the time of transfer. Notably, the proportion of embryos recovered decreased (P = 0.03) as more embryos were transferred, particularly at later stages (Day 14, P < 0.01). The average length of embryos decreased by approximately 5% for every additional embryo transferred (P < 0.0001). These effects may be linked to embryonic migration. Embryo mortality inherent to the embryo during the second week of pregnancy was 24%. Additionally, 9% of Day 14 embryos were of inferior quality, as they did not contain an epiblast. Combining embryo and recipient causes but excluding infection effects, embryonic loss of IVP embryos during the second week of pregnancy amounted to 26% (heifers) or 34% (parous dairy cows). The length of embryos doubled every day between Days 9 and 16, with a 4.4-fold range in sizes representing two thirds of the variation in length. Embryos retrieved from heifers were twice the size of those incubated in parous cows (P < 0.0001), indicating faster embryonic development/trophoblast proliferation in heifers. Whereas season did not affect embryo recoveries, length was lower (50%) in winter (winter-autumn, P < 0.05; winter-spring, P < 0.001). Lastly, transuterine migration in cattle, when transferring multiple embryos, commenced at Day 14 (4%) and had occurred in all recipients by Day 16 (38% of embryos found contralaterally).     

Nonsurgical embryo transfer in the common marmoset monkey

Abstract: A technique for nonsurgical embryo transfer in common marmosets was developed. Transfers were either synchronous (ST) or asynchronous (AT). Synchronous transfers (embryo donor and the embryo recipient ovulated on the same day) were performed 5 to 8 days post-ovulation. Asynchronous transfers (embryo donor had ovulated at least 2 days before the embryo recipient) were performed when the recipient was 2 to 4 days post-ovulation (donor was 6 to 8 days post-ovulation). Four pregnancies from nine transfers (44%) were established by AT, and three pregnancies were carried to term. Only 1 of 11 transfers (9%) from ST resulted in a pregnancy, which was lost by Day 40 of gestation. Significantly more infants were born from AT (6 infants from 17 embryos; 35%) than from ST (0 infants from 22 embryos; 0%; p<0.005). This technique allows experimental analysis of primate postimplantation development and provides a tool for conservation of endangered Callithrichid species.     

The GnRH antagonist acyline prevented ovulation, but did not affect ovarian follicular development or gestational corpora lutea in the domestic cat

Two experiments were conducted to investigate the effects of the GnRH antagonist acyline (330 μg/kg, given sc) on ovarian follicular development and ovulation, as well as on pregnancy maintenance in domestic cats. In the first experiment, seven queens in proestrus (total of 24 proestrus periods), were randomly assigned to treatment with either acyline (ACY; n = 17) or a placebo (PLC; n = 7). All queens were mated with a fertile tomcat. In the ACY and PLC groups, cessation of estrus occurred (mean ± SEM) 7.0 ± 1.3 and 7.0 ± 1.7 d after treatment (P > 0.1), ovulation occurred in 2 of 17 and all seven estrus periods (P < 0.05), and pregnancy rates were 1 of 16 and 7 of 7 (P < 0.05), respectively. In the ACY and PLC groups, intervals from treatment to the onset of the ensuing proestrus were 18.4 ± 1.7 and 120 ± 17.2 d. In the second experiment, 14 pregnant queens were randomly allocated, according to their mating date, to treatment with acyline in early pregnancy (from 20 to 25 d, n = 3), mid pregnancy (from 26 to 45 d; n = 4), late pregnancy (> 45 d; n = 3), or injection of a placebo in early (n = 1), mid (n = 2), or late pregnancy (n = 1). Ultrasonographic assessments of the uterus were done every second day for 2 wk post treatment, and serum progesterone (P₄) concentrations were determined before treatment, and at 7 and 14 d after treatment. No pregnancies were prematurely terminated and post-treatment P₄ concentrations did not differ among treatment groups (P > 0.1). In conclusion, in the domestic cat, GnRH withdrawal by acyline prevented ovulation when given in early follicular phase (proestrus), but did not significantly affect luteal function during pregnancy.     

Effect of progesterone supplementation in the first week post conception on embryo survival in beef heifers

Progesterone is essential for establishment and maintenance of pregnancy in mammals. The objective of this study was to examine the effect of elevating progesterone during the different physiological stages of early embryo development on embryo survival. Estrus was synchronized in cross-bred beef heifers (n = 197, ∼2-years old) and they were inseminated 12-18 h after estrus onset (=Day 0). Inseminated heifers were randomly assigned to 1 of 3 treatments: (1) Control, n = 69; (2) progesterone supplementation using a Controlled Internal Drug Release Device (CIDR) from Day 3 to 6.5, n = 64; or (3) progesterone supplementation using a CIDR from Day 4.5 to 8, n = 64. Body condition (BCS) and locomotion scores (scale of 1-5) were recorded for all animals. Animals with a locomotion score ≥4 (very lame) were excluded. Embryo survival rate was determined at slaughter on Day 25. Conceptus length and weight were recorded and the corpus luteum (CL) of all pregnant animals was dissected and weighed. Supplementation with exogenous progesterone increased (P < 0.05) peripheral progesterone concentrations, but did not affect embryo survival rate compared with controls. Mean CL weight, conceptus length and conceptus weight were not different between treatments. There was a positive relationship (P < 0.04) between the increase in progesterone concentrations from Days 3 to 6.5 and embryo survival rate in treated heifers and a similar trend existed between the increase from Days 4.5 to 8 (P < 0.06). There was also a positive relationship (P < 0.05) between the progesterone concentration on Day 6.5 and the embryo survival rate in treated heifers. A direct correlation was seen between locomotion score and embryo survival rate, with higher (P < 0.05) early embryo survival rates in heifers with a lower locomotion score. In conclusion, supplementation with progesterone at different stages of early embryo development increased peripheral progesterone concentration and resulted in a positive association between changes in progesterone concentration during the early luteal phase and embryo survival rate. Supplementation with progesterone had no effect on either CL weight or conceptus size in pregnant animals. Lameness had a significant negative effect on early embryo survival.     

Immunization against inhibin enhances both embryo quantity and quality in Holstein heifers after superovulation and insemination with sex-sorted semen

The objective was to investigate the feasibility of improving embryo yield in superovulated cows following insemination with sex-sorted semen by prior immunization against inhibin. Twenty-eight heifers were allocated into three groups: High (n = 10), Low (n = 10), and Control (n = 8). The High group received one primary (1 mg) and two booster (0.5 mg) vaccinations (28-d intervals) with a recombinant inhibin α-subunit in 1 mL of white oil adjuvant, whereas the Low group received half that dose, and the Control group received only adjuvant. After the last immunization, all heifers underwent a standard superovulation treatment (decreasing doses of pFSH for 4 d), followed by two AI with 2 × 10⁶ sex-sorted semen after the onset of estrus. Inhibin-immunized heifers had higher (P < 0.01) plasma antibody titres, and an earlier onset of estrus (P < 0.05) than Control heifers. The total number of embryo/ova, transferable, and grade 1 embryos in the High group (15.4 ± 1.9, 5.7 ± 0.7, and 3.8 ± 1.0, respectively) was significantly greater than that of the Control group (9.1 ± 1.2, 3.1 ± 0.5, and 0.6 ± 0.2), but was intermediate (P > 0.05) in the Low group (13.0 ± 2.3, 4.4 ± 0.7, and 1.2 ± 0.3). There were no significant differences among groups in number of unfertilized ova and degenerated embryos. The High group also had higher (P > 0.05) plasma progesterone concentrations on the day of embryo collection. In conclusion, immunization against inhibin improved both embryo quantity and quality following superovulation and insemination with sex-sorted semen.     

Effect of MEM vitamins and forskolin on embryo development and vitrification tolerance of in vitro-produced pig embryos

The aims of this study were (1) to determine the effect of in vitro maturation (IVM) medium supplementation with MEM vitamins on in vitro embryo development and sensitivity to vitrification of Day 6 blastocysts and (2) to evaluate whether the addition of forskolin to in vitro culture (IVC) medium enhances blastocyst survival following Super Open Pulled Straw (SOPS) vitrification. Cumulus–oocyte complexes (COCs; n = 4000) were matured with 0.0% or 0.05% (v/v) MEM vitamins. After 44 h of IVM, the oocytes were in vitro fertilized, and presumptive zygotes were cultured. At Day 5 of IVC, embryos from both experimental groups were cultured for 24 h with 0 or 10 μM forskolin, achieving a 2 × 2 factorial design. The blastocyst formation rate was assessed on Day 6, and subsets of samples from the four experimental groups were vitrified (n = 469) or kept fresh (n = 546). Fresh and vitrified-warmed blastocysts were cultured for 24 h prior to embryo survival and total blastocyst cell number assessment. The MEM vitamins increased (P < 0.001) the blastocyst formation rate at Day 6, but they did not affect embryo survival after vitrification. In contrast, the addition of forskolin to the culture medium enhanced (P < 0.05) the blastocyst vitrification tolerance. The total blastocyst cell number was similar among the groups. In conclusion, supplementation with 0.05% MEM vitamins improved the blastocyst formation rate, and the addition of 10 μM forskolin to the culture medium increased survival in Day 6 in vitro-produced blastocysts after SOPS vitrification.     

Reproductive toxicologic evaluations of Bulbine natalensis Baker stem extract in albino rats

The effects of oral administration of aqueous extract of Bulbine natalensis Baker stem at daily doses of 25, 50, and 100 mg/kg body weight on the reproductive function of Wistar rats were evaluated. The indices of mating and fertility success as well as quantal frequency increased after 7 days of treatment in all the dose groups except the 100 mg/kg body weight group. The number of litters was not statistically different (P > 0.05) from the control. Whereas the absolute weights of the epididymis, seminal vesicle, and prostate were not affected, that of the testes was significantly increased. The epididymal sperm count, motility, morphology, and viscosity were not different from the control after 7 days of treatment. The male rat serum testosterone, progesterone, luteinizing hormone, and follicle-stimulating hormone significantly increased in the 25 and 50 mg/kg body weight groups, whereas the estradiol concentration decreased significantly at all the doses. The extract dose of 100 mg/kg body weight decreased the serum testosterone and progesterone levels in male rats. The prolactin concentration was not affected by all the doses. All the indices of reproduction, maternal, embryo/fetotoxic, teratogenic, and reproductive hormones in the female rats were not statistically different from that of their control except the resorption index, which increased at the dose of 100 mg/kg body weight of the extract. Histologic examination of the cross section of rat testes that received the extract at all the doses investigated revealed well-preserved seminiferous tubules with normal amount of stroma, normal population of spermatogenic and supporting cells, as well as normal spermatocytes within the lumen. The results revealed that the aqueous extract of Bulbine natalensis stem at doses of 25 and 50 mg/kg body weight enhanced the success rate of mating and fertility due to increased libido as well as the levels of reproductive hormones in male rats. The absence of alterations in the reproductive parameters of female rats at doses of 25 and 50 mg/kg body weight of Bulbine natalensis stem extract suggest that the extract is “safe” for use at these doses by females during the organogenic period of pregnancy, whereas the extract dose of 100 mg/kg body weight portends a negative effect on some reproductive functions of male and female rats.     

Effects of treatment with a prostaglandin analogue on developmental dynamics and functionality of induced corpora lutea in goats

The aim of this study was to compare morphological and functional features of spontaneous and induced corpora lutea (CLs) in goats. Fourteen adult and cycling Anglo Nubian goats (Argentina) were randomly allocated to two groups: Group N (n = 7) included goats with natural spontaneous oestrus and Group PG (n = 7) included does in which oestrus was synchronized by the administration of two i.m. cloprostenol doses, 10 days apart. In both groups, oestrous behaviour was checked twice daily (Day of oestrus = Day 0) and daily transrectal ultrasonographies were performed for evaluating CLs and follicles dynamics through the complete subsequent oestrous cycle; the luteal activity was determined directly, in terms of progesterone (P4) secretion, and indirectly, by assessing effects of CL on follicular dynamics. All goats exhibited oestrous behaviour and ovulation without differences in ovulation rate (N: 1.67 ± 0.2, PG: 2.0 ± 0.1). The total luteal tissue area showed linear growth from Day 4 to Day 15 of oestrous cycle in all goats, but the developmental dynamics differed between groups, treated goats had larger area (P < 0.01). Plasma P4 concentrations also increased from Day 0 to Day 15 in all the does; however, from Day 5 to Day 15, treated does had a lower concentrations than the untreated group (P < 0.001). There were differences in the development of follicular waves between groups; assessment of size-distribution showed that treated group had a higher number of small and larger follicles (P < 0.05). The largest follicles recorded in treated goats had a higher maximum diameter both at the first (PG: 7.6 ± 0.8 mm; N: 4.9 ± 0.7 mm, P < 0.05) and second follicular waves (PG: 6.3 ± 1.4 mm; N: 5.0 ± 0.4 mm, P < 0.05) and a longer growth phase during the second wave (PG: 6.5 ± 1.7 days; N: 4.6 ± 0.7 days, P < 0.05), coincident with the period of maximal luteal secretion. In conclusion, synchronization of oestrus and ovulation by the administration of a prostaglandin analogue causes differences in developmental dynamics and functionality of induced corpora lutea when compared to natural spontaneous ovulation.     

Effect of addition of hyaluronan to embryo culture medium on survival of bovine embryos in vitro following vitrification and establishment of pregnancy after transfer to recipients

Two experiments were conducted to determine whether addition of hyaluronan to culture medium could improve survival of bovine embryos after vitrification or following embryo transfer. In Experiment 1, embryos were produced in vitro and cultured for 7 days in modified synthetic oviductal fluid (SOF) containing one of four concentrations of hyaluronan (0, 0.1, 0.5, or 1 mg/mL), with or without 4 mg/mL of bovine serum albumin (BSA). On Day 7 after insemination, blastocysts and expanded blastocysts were vitrified using open-pulled straws. At a concentration of 1 mg/mL, hyaluronan increased (P < 0.05) the percentage of oocytes that were blastocysts and re-expansion rate at 24 h after warming. At 0.5 mg/mL, hyaluronan tended (P < 0.10) to increase re-expansion rate at 48 h after warming and increased (P < 0.05) embryo hatching rate at 24 and 72 h. Treatment with BSA caused a slight reduction in cleavage rate (P < 0.05), but only for cultures containing hyaluronan (BSA × hyaluronan, P = 0.10), an increase in the percentage of oocytes that became blastocysts (P < 0.001), and a reduction in re-expansion rates (P < 0.001) and hatching rates (P < 0.05 or P < 0.01) at all times examined. In Experiment 2, embryos were produced in vitro and cultured in modified SOF containing 4 mg/mL BSA, with or without 1 mg/mL hyaluronan. At 159-162 h after insemination, grade 1 morula, blastocysts and expanded blastocysts were harvested for embryo transfer. Harvested embryos were transferred individually to lactating Holstein recipients with a palpable corpus luteum on Day 7 after presumptive ovulation. There was an interaction (P < 0.05) between hyaluronan and embryo stage on pregnancy rate. Recipients that received morula and blastocyst stage embryos treated with hyaluronan had a higher pregnancy rate than recipients that received control embryos of the same stage. There was no effect of hyaluronan on pregnancy rates of recipients that received expanded blastocysts. In conclusion, addition of hyaluronan to embryo culture enhanced blastocyst yield, improved survival following vitrification, and enhanced the post-transfer survival of fresh morula and blastocyst stage embryos.     

Effects of embryo size at transfer (whole versus demi) and early pregnancy progesterone supplementation on embryo growth and pregnancy-specific protein bovine concentrations in recipient dairy heifers

The objectives of this study were to evaluate embryonic size and survival, plasma progesterone (P4) and pregnancy-specific protein bovine (PSPB) concentrations in early pregnancies (n = 99) following the transfer of one whole (n = 66) or one demi (n = 33) embryo to recipient virgin dairy heifers. The experiment was designed to evaluate the fixed effects of embryo size at transfer (whole or demi embryo) on Day 7 of the estrous cycle (Day 0 = estrus) and P4 supplementation between Days 7 to 19 through an intravaginal device (yes or no) on plasma P4 and PSPB concentrations and on embryo measurements. Plasma P4 concentrations were measured by RIA on Days 0, 7, 14, 19, 21, 25, 35, 42, 49, 56 and 63 of pregnancy and, PSPB concentrations were measured by ELISA on Days 7, 21, 25, 35, 42, 49, 56 and 63. The presence of an embryonic vesicle was detected on Day 25, embryonic/fetal movements and heartbeat were evaluated on Days 42 and 63 and embryo measurements [crown-rump length (CRL) and width at mid body] were obtained on Day 42 through ultrasonography.In non-supplemented pregnancies, Day 42 whole embryos had higher (P < 0.05) CRL and width than demi embryos, but the difference averaged only 1 to 2 mm. In P4 supplemented pregnancies, whole and demi embryos attained a similar size on Day 42 of pregnancy. Embryo size at transfer, early exogenous P4 supplementation and their interactions had no effects (P > 0.05) on plasma P4 concentrations. However, the post-hoc LSD evaluation showed that plasma P4 concentrations on Day 25 were higher (P < 0.001) in whole than in demi embryo derived pregnancies and, that exogenous P4 supplementation increased (P < 0.05) plasma P4 concentrations on Day 19 of pregnancy. The plasma PSPB detection rate on Days 7 to 63 of pregnancy was similar in pregnancies resulting from the transfer of whole and demi embryos. From a total of 93 recipients remaining pregnant until Day 63, plasma PSPB was constantly undetectable on Day 7, was detected in 4% of Day 21 samples, 41% of Day 25, 95% of Day 35, 96% of Day 42, 99% of Day 49 and in 100% of samples of Days 56 and 63. Concentrations of PSPB increased (P < 0.05) from Days 21 to 42 and from Days 56 to 63, with a plateau between Days 42 to 56. Demi embryo pregnancies had higher (P < 0.05) plasma PSPB concentrations on Days 35 and 42 than whole embryo pregnancies. Progesterone supplementation had a positive effect (P < 0.01) on PSPB concentrations from Days 35 to 63. Concentrations of PSPB were similar in non-supplemented whole and demi embryo pregnancies from Days 7 to Day 63. In contrast, in supplemented recipients, demi embryo pregnancies had higher (P < 0.05) PSPB concentrations on Days 25 to 42 than whole embryo pregnancies. No significant correlation was found between P4 and PSPB concentrations or between the concentrations of these hormones and embryonic measurements on Day 42. In conclusion, demi embryos experienced a compensatory growth until Day 42 of pregnancy, attaining a similar size to that of whole embryos and originating conceptuses producing similar plasma PSPB concentrations to those of whole embryo derived conceptuses. Embryonic growth and conceptus secretion of PSPB were positively stimulated by early pregnancy exogenous P4 treatment.     

Morphometric analysis of terminal villi and gross morphological changes in the placentae of term idiopathic intrauterine growth restriction

The aim of this study is to compare the gross morphology of the placentae and the morphometry of terminal villi and terminal villous capillaries in pregnancies complicated by idiopathic intrauterine growth restriction (IUGR) with those of normal pregnancies. 75 placentae were collected between April 2010 and March 2011. 50 placentae were associated with idiopathic IUGR and 25 were from controls. Insertion of cords, placental weights and diameters were noted. Hematoxylin and eosin-stained wax sections were analyzed stereologically. Growth of terminal villi and fetal capillaries was assessed by estimating total and mean surface areas. Villous capillarization was monitored using capillary:villus surface ratio. Measurements were done using image analysis system. In comparison with the control group, idiopathic IUGR placentae are significantly smaller (p = 0.000) and lighter (p = 0.000). In majority of IUGR (68%) and control (60%) cases, eccentric insertion of cord is noted. In idiopathic IUGR group, there is a significant decrease in the total areas of both terminal villi (p = 0.048) and their capillaries (p = 0.000) and a significant decrease in number of both terminal villi (p = 0.000) and their capillaries (p = 0.001), also, capillarization index is significantly smaller (p = 0.038). Idiopathic IUGR is associated with reduced growth of placental terminal villi and fetal capillaries and this is accompanied by changes in measures of villous capillarization as compared with those of control placentae. Further investigations of idiopathic IUGR placentae are necessary, especially considering the histopathological changes that could affect the fetomaternal exchange, with a note that strict distinction should be made between idiopathic and nonidiopathic IUGR placentae.     

Variation in gestation length among captive reindeer (Rangifer tarandus tarandus)

An estimated 90% of reindeer females are mated in a 10- to 21-d interval and give birth in an equally synchronized manner. Reported gestation length in reindeer is highly variable (range, 203 to 240 d), almost twice the reindeer estrous cycle length. Previously, we identified a significant, negative relationship between gestation length and conception date in a small group of reindeer. In the current study, the negative relationship was investigated in a switchback design, where reindeer were divided into two groups synchronized for early and late mating over a 2-yr trial. Regression analysis of 11 paired observations produced a negative (P < 0.001) association between gestation length and conception date (slope = − 0.31). Dam weight at breeding and prior to parturition, calf birth weight, and calf sex were not significant variables in the regression. Regression analysis of a larger data set from two University of Alaska Fairbanks reindeer herds, where conception date (verified by systemic progesterone) and gestation length were recorded (historical data set), supported previous conclusions (n = 70; slope = −0.37; P < 0.001). Although the calf sex ratio did not differ with gestation length, there was a positive relationship (r² = 0.19; P = 0.014) between male birth weight and gestation length in the larger data set. The negative relationship between conception date and gestation length enhanced calving synchrony, though the limits of gestation plasticity and underlying mechanisms are not clear. The potential role of photoperiod on early embryonic development is discussed.     

Early pregnancy diagnosis and embryonic and fetal development using real time B mode ultrasound in goats

Real time B mode ultrasound scanning was used for the diagnosis of early pregnancy and monitoring of embryonic and fetal development in goats. Estrus in 8 cyclic cross-bred does was synchronized using a single injection of PGF2α followed by mating with a buck. All the does were subjected to weekly transrectal (from day 14 of mating) and transabdominal ultrasonography (from day 28) up to the 4th month of gestation and once during the 5th month of gestation. Of the 8 animals, 6 does were confirmed pregnant. The embryonic vesicle was detected on day 21 and day 28 and the embryo proper on day 28, day 35 using the transrectal and transabdominal methods, respectively. Heart beat was observed as early as day 21 of gestation and it was recordable by day 28 using the transrectal approach. However, via transabdominal scanning the detection and recording of the heart beat was possible only on day 35 of gestation. Singles and twins were differentiated on day 35 and day 42 by the transrectal and transabdominal approaches, respectively. Placentomes as a circular ‘C’ shape structure was detectable on day 42 with an average diameter of 0.97 cm, using the rectal probe. Using the abdominal probe, placentomes were detectable on day 50 (average diameter of 1 cm of gestation). Skeletal structures such as the skull, rib cage and vertebral column were first viewed on day 56 in both approaches. A significant (P < 0.001) high correlation (r = 0.99) was recorded between the gestational age and head, thoracic and placentome parameters. It may concluded that the earliest identification of an embryonic vesicle (day 21) by transrectal scanning and confirmation of its viability by fetal heart beat (day 28) were the most reliable ultrasonographic features for early pregnancy diagnosis in goats. Transabdominal ultrasonographic measurements of placentome, head and thoracic diameter may be of use in the assessment of fetal age.     

The evaluation of corpus luteum blood flow using color-flow Doppler ultrasound for early pregnancy diagnosis in bovine embryo recipients

The objective of this experiment was to evaluate corpus luteum blood flow (CLBF) as an early indicator of pregnancy status in bovine embryo recipients. Fifty crossbred beef cows were submitted to embryo transfer on Day 7 after estrus. On Days 7, 11, 13, 15, 17, 19, 21, 26, 33, and 40, a blood sample was taken, the CL examined using a color-flow Doppler ultrasound scanner, and video was recorded of each scanning session. Ultrasound data were grouped by the first day progesterone concentrations were <1 ng/mL (indicating early embryo loss, EEL) through Day 21 (EEL-17, n = 3; EEL-19, n = 9; EEL-21, n = 3), absence of an embryo on Days 26, 33, or 40 (late embryo loss; LEL; n = 12), or remained pregnant (P; n = 23). The first decrease in CLBF of EEL-17, EEL-19, and EEL-21 cows compared to P cows occurred on Days 17, 19, and 21, respectively (P < 0.05). There was no difference in CLBF between LEL and P cows on Days 17, 19, and 21. Six evaluators diagnosed pregnancy from randomized video clips on Days 17, 19, and 21. Evaluators made more (P < 0.004) correct diagnoses on Day 19 than Day 17. Sensitivity (82.9 ± 10.1%) was not affected by day. From Days 17 to 19, diagnostic specificity increased (P = 0.046) from 43.2 ± 3.0 to 54.3 ± 3.0% but remained unchanged thereafter. Due to low specificity and sensitivity, evaluation of CLBF alone was insufficient for early pregnancy diagnosis.     

The relationship between consecutive pregnancies in Thoroughbred mares. Does the location of one pregnancy affect the location of the next, is this affected by mare age and foal heat to conception interval or related to pregnancy success

Delayed uterine involution is a major cause of early reproductive failure in mares. Involution is affected by mare age, and foaling to covering interval. Involution rates vary between the previously non-gravid horn (PNGH), which recovers the quicker, and the previously gravid horn (PGH). Location of a pregnancy and its likely success may, therefore, be affected by its location relative to the previous pregnancy. This study aimed to determine: (i) the location of concepti in consecutive pregnancies; (ii) whether this varies with mare age or foaling to conception interval; (iii) whether location in relation to the previous pregnancy affects success. 1383 Thoroughbred mares were monitored by ultrasonic scanning during oestrus and early pregnancy. Significantly (p < 0.01) more pregnancies were located in the PNGH (79.2%) than the PGH (20.8%). The number of pregnancies in PGH significantly increased with mare age (p < 0.01) and foaling to conception interval (p < 0.05). Significantly (p < 0.001) more pregnancies located in the PGH (16.5%) failed, than those in the PNGH (4.6%). It can be concluded that most pregnancies locate in the PNGH where their chances of success are greatest. The larger number of pregnancies locating in the PGH in older mares and those with shorter foaling to conception intervals may in part account for the higher conceptus mortality rates in such mares. Hence breeding older mares on alternate years and maximising foaling to conception interval may improve reproductive success. Alternatively termination of pregnancies located in the PGH followed by timely recovering may be justifiable as might ET in older mares covered close to foaling.