The extent and timing of prenatal loss in gilts

The potential litter size of gilts that is based on the ovulation rate is much higher than the actual litter size, which depends on the fertilization rate and subsequent prenatal mortality. Prenatal mortality is divided into embryonic mortality (before Day 30) and fetal mortality (after Day 30). Prenatal loss includes both fertilization failure and prenatal mortality. Crossbred gilts (n = 149) were bred at the first observed estrus after being exposed to the boar at 200 days of age. Time of the first insemination after estrus detection was determined by measurement of vaginal conductivity using a Walsmeta meter. A second insemination was administered either 8 or 16 hours later. Artificial insemination with fresh semen (0 to 3 days old) was used throughout the experiment. Gilts were slaughtered on Day 3 (n = 26), Day 10 (n = 42), Day 30 of gestation (n = 45) or they were allowed to farrow (n = 36). Gilts slaughtered on Day 3 were used to estimate the fertilization rate. Gilts slaughtered on Day 10 and Day 30 were used to calculate embryonic mortality, while fetal mortality was calculated from the gilts that farrowed. The mean (+/-SEM) number of corpora lutea (CL) was 13.15+/-0.46, 13.36+/-0.37 and 12.97+/-0.39 for gilts slaughtered at Days 3, 10 and 30, respectively (P>0.05), and the mean (+/-SEM) number of normal embryos recovered was 11.12+/-0.69, 9.46+/-0.55 and 9.33+/-0.58, respectively. Litter size at parturition was 9.10+/-0.54. There was a significant difference between the number of normal embryos on Day 3 and Day 30 (P=0.05) and also between the number of normal embryos at Day 3 and the number of piglets at term. Ninety percent of the ova were recovered at Day 3. The fertilization rate was calculated either 1) assuming that unrecovered ova had a similar fertilization rate as the recovered ova (FRER=94.5+/-2.0%) or 2) assuming that unrecovered ova were unfertilized (FROR=84.5+/-2.5%). It was concluded that FRER was a more accurate estimation of the fertilization rate. Based on this fertilization rate, embryonic mortality between Day 3 and Day 10 was 20.8+/-8.3%, with an additional 12.5+/-7.1% loss between Day 10 and Day 30, when all gilts were included (P = 0.308). Thus the total prenatal loss, including fertilization failure, up to Day 10 was 26.3% and to Day 30 it was 38.8%. Fetal mortality was 2.2%, giving a total prenatal mortality (excluding fertilization failure) of 35.5% and a prenatal loss of 41%. Most of the prenatal loss was due to embryonic mortality. In those gilts that remained pregnant most of the embryonic loss occurred before Day 10 (19.0+/-6.3%; P=0.003). There was no further loss between Day 10 and 30 of pregnancy. There was a significant difference between the loss from Day 3 to Day 10 compared with the loss from Day 10 to Day 30 (P=0.05); therefore, most of the embryonic loss in pregnant gilts occurred before Day 10. Since fetal mortality was 3.2+/-6.3%, most of the prenatal loss was due to embryonic mortality.     

The effects of low levels of PMSG and flush feeding upon embryonic survival in gilts

This experiment was designed to measure the effects of a dose of PMSG (600 IU) or a five-day flush feeding, singly or in combination, on ovulation rate, early embryonic survival, litter size at term and plasma hormone levels during early gestation in 64 Duroc gilts. They were assigned to four treatments, with half of each treatment killed on day 25 of pregnancy and the other half continuing to term.Although treatment with 600 IU of PMSG increased the number of ovulations compared to the controls (P < .05), it failed to increase the number of viable embryos at day 25 of gestation (P > .05). Flush feeding for five days before breeding yield similar results. Fewer embryos were present at day 25 of gestation in those animals receiving a combination $\text{PMSG}$/flush feeding treatment than in any other treatment group (P < 0.01). At day 25 of gestation, the average prenatal survival rate was 71% for all treatments. At parturition, there were no differences between treatment groups in the average number of pigs born per litter; nor were there any differences present for the average number of pigs weaned per litter at four weeks postpartum. Plasma progesterone levels at the onset of treatment, mating and during early gestation were found to be similar among the four treatment groups. Plasma estradiol-17β levels were also similar among the four groups. Based on this experiment, it would appear that factors other than the mere number of viable eggs shed determines the litter size at parturition in swine.     

Synchronization of oestrus in gilts with altrenogest: effects on ovulation rate and foetal survival

Previous studies have shown that use of altrenogest resulted in a high rate of fertility and increased litter size compared with controls under conditions of practical pig production. The present study was designed to evaluate whether ovulation rate and/or foetal survival were increased by altrenogest using crossbred gilts derived from one herd (n = 227) and introduced in the same piggery over 12 months. Each gilt was allocated to a treated group (n = 103) receiving an individual daily dose of 20 mg of altrenogest for 18 days in its feed or a control group (n = 124) after puberty had been diagnosed, (197 ± 1 day; mean ± SEM). They were inseminated (double AI) at the second induced or natural oestrus. Pregnancy was diagnosed by ultrasonography at Days 22 and 42 post-insemination in the absence of return to oestrus. Pregnant gilts were slaughtered at 48 ± 3 days of pregnancy following the second examination. The number of living and dead foetuses were recorded before uterine contents (foetuses and placentae) were weighed and the number of corpora lutea (CL) per ovary counted.Precise synchronization of oestrus was observed after the end of the progestogen administration, with 93% of the gilts in oestrus by Days 5 to 7. For the controls, the interval from first to second oestrus ranged from 17 to 25 days in 93% of the control gilts. The pregnancy rate was 89.3% for treated gilts and 77.4% for controls (P < 0.05). The ovulation rate was increased by the treatment (15.4 ± 0.3 vs 14.6 ± 0.3; mean ± SEM, P < 0.02). Although the altrenogest group had more foetuses (11.1 vs 10.6), this difference was not significant (P > 0.14). The percent of foetal survival was similar in both groups (64.9%; P > 0.27). The foetal and placental weights differed only between dams and increased with stage of gestation. The increase in litter size through feeding altrenogest was associated with an increased ovulation rate.     

Effect of hCG administration at the onset of oestrus on early embryo survival and development in Meishan gilts

A study was designed to advance the time of the ovulatory luteinizing hormone (LH) surge in Meishan gilts by human chorionic gonadotrophin (hCG) administration at the onset of oestrus and assess the effect on embryo survival and development. Twelve Meishan gilts were observed six times daily for oestrous behaviour and bred at 24, 36 and 48 h after observed oestrous onset. Six of those gilts were administered an ovulatory dose of hCG (500 IU) at observed oestrous onset. Blood samples were collected at oestrous onset (Day —2) and on Days 0, 2, 6, 9, 13, 16, 20, 23, 27 and 30 of gestation. All gilts were slaughtered on Day 30 of gestation and embryo survival and conceptus development assessed. Ovulation rate did not differ between control and hCG treated gilts (18.5 and 17.7 respectively; P>0.1) while the number of live conceptuses per gilt (17.2 and 12.8 respectively; P<0.08) and embryo survival rate (92.1 and 75.8% respectively; P<0.1) both tended to be reduced by the hCG treatment. Placental weight (17.2 and 23.1 g; P<0.01) was significantly increased in hCG treated gilts, while embryo weight (1.2 and 1.4 g; P<0.06) and placental length (42.8 and 47.2 cm; P<0.07) both tended to be increased in hCG treated gilts. Crown rump length (P>0.1) and allantoic fluid volume (P>0.1) did not differ between the treatment groups. Serum progesterone concentrations did not differ with treatment overall (P>0.1) but were significantly elevated (P< 0.05) at 48 h postoestrus in the hCG treated gilts compared to control gilts. Overall, these results indicate that advancing the time of the LH surge to oestrous onset, as in European breeds, compromised embryo survival and suggests that the longer time interval between oestrous onset and ovulation is important for the high rate of embryo survival in the Meishan pig.     

The relationship between ovulation rate and embryonic survival in gilts

Norwegian Landrace gilts were inseminated on the second day of their second oestrus and slaughtered 28 to 34 days after insemination. The number of corpora lutea (ovulation rate) and normal embryos was counted and the embryonic survival rate was calculated for the 306 pregnant gilts. Mean (+/-S.D.) ovulation rate, number of normal embryos and embryonic survival rate were 14.17+/-2.48, 10.55+/-3.30 and 74.8%+/-20.7%, respectively. The significant (P<0.001) curvilinear regression of embryonic survival rate on ovulation rate gives a maximum embryonic survival rate at 13.2 ovulations. Increased ovulation rate gives increased number of normal embryos up to 18.1 ovulations. Ovulation rate should be considered when assessing factors affecting embryonic survival in pigs.     

The cellular effects of estrogens on neuroendocrine tissues

Estrogen action on sensitive neurons in the rat diencephalon has been studied by morphologic techniques; evidence of estrogen action at every level is presented, including tracts, cells, circuitry and subcellular organelles. The demonstration in the arcuate nucleus of estrogen-induced synaptic remodelling, estrogen-induced postsynaptic membrane phenotypes, changes in intracellular membranes and rapid estrogen actions on neuronal endo-exocytosis indicates that cellular estrogen actions may underlie the neuronal control of reproduction.     

Donor and recipient genotype and heterosis effects on survival and prenatal growth of transferred mouse embryos

Reciprocal embryo transfers amongst two inbred strains (C3HeB/FeJ and SWR/J) and their F1 cross (C3SWF1) were used to examine donor and recipient genotype and heterosis effects on survival and prenatal growth of mouse embryos. Among inbred strains, significant recipient genotype effects were detected for both embryo survival (P less than 0.01) and prenatal growth (P less than 0.05), while no donor genotype effects were observed. The recipient effect on overall embryo survival was due to a higher proportion of C3H recipients maintaining pregnancy to term than SWR recipients (P less than 0.01), rather than survival within litters. Irrespective of their own genotype, embryos developing in C3H uteri achieved larger body weights (P less than 0.01) and longer tail lengths (P less than 0.05) at birth than did embryos developing in SWR uteri. Recipient heterosis was not significant, while donor heterosis was significant for prenatal growth traits (P less than 0.001).     

Gonadotropin and prolactin secretion following intraventricular administration of morphine in gilts

The effects of central nervous system administration of morphine on secretion of luteinizing hormone (LH), follicle-stimulating hormone, and prolactin were investigated in ovariectomized gilts stereotaxically implanted with lateral ventricular cannulas. In Experiment 1, mean serum LH and follicle-stimulating hormone concentrations and serum LH pulse frequency were unaffected by artificial cerebrospinal fluid administration (P greater than 0.1), but decreased (P less than 0.01) in 8 of 11 gilts when 500 micrograms of morphine were given 3 hr later. Serum LH pulse amplitude was unaffected (P greater than 0.1) by cerebrospinal fluid or morphine injection. In Experiment 2, luteinizing hormone concentrations decreased (P less than 0.0001) and prolactin concentrations increased (P less than 0.0001), but follicle-stimulating hormone concentrations did not change (P greater than 0.1) after 500 micrograms of morphine. Gonadotropin responses to 10 micrograms of gonadotropin-releasing hormone, given 2 hr after intraventricular injection, were similar (P greater than 0.1) for morphine- and cerebrospinal fluid-treated gilts. These results indicate that morphine inhibits LH secretion at the level of the central nervous system, and are consistent with the concept that endogenous opioid peptides participate in the regulation of gonadotropin and prolactin release in pigs.     

Increased ovulation rate in gilts after oral administration of epostane

In Phase I of this study to enhance ovulation rate and hence litter size, gilts received 0 (sham control), 0.625, 1.25, 2.5 or 5.0 mg epostane/kg body weight on Days 10, 11 and 12 of the oestrous cycle (5 gilts/group). After epostane treatment, plasma progesterone concentrations were reduced (P less than 0.01) in a dose-related manner, % progesterone decline = 21.30 x square root of (dose) + 10.45, R2 = 0.70, but recovered to pretreatment levels by 24 h. In Phase II the effects of epostane on ovulation rate and litter size were tested at two study centres. At each centre 108 gilts were treated with the same doses of epostane as used in Phase I and the doses were given for 7 days (Days 15-21) or 12 days (Days 10-21) during the first oestrous cycle. Gilts were inseminated twice during the oestrus after treatment and were slaughtered 30 days later. Mean (+/- s.d.) ovulation rate was 16 +/- 2.7 (N = 8) and 21 +/- 4.0 (N = 61) for control and epostane-treated gilts in Centre A and 12 +/- 2.4 (N = 5) and 17 +/- 3.8 (N = 55) respectively in Centre B (P less than 0.01 for both) and was dose related (ovulation rate = 3.38 x square root of (dose) + 16.17, R2 = 0.31). The effects of 7- or 12-day epostane treatment on ovulation rate were not different (P greater than 0.05), indicating that effects of treatment after Day 14 of the oestrous cycle are most important to subsequent ovulation frequency.(ABSTRACT TRUNCATED AT 250 WORDS)     

Genotoxic effects of estrogens

Estrogens are associated with several cancers in humans and are known to induce tumors in rodents. In this review a mechanism of carcinogenesis by estrogens is discussed which features the following key events: (1) Steroid estrogens are metabolized by estrogen 2- and 4-hydroxylases to catecholestrogens. Target organs of estrogen-induced carcinogenesis, hamster kidney or mouse uterus, contain high levels of estrogen 4-hydroxylase activity. Since the methylation of 4-hydroxyestradiol by catechol-O-methyltransferase is inhibited by 2-hydroxyestradiol, it is proposed that a build up of 4-hydroxyestrogens precedes estrogen-induced cancer. (2) The catecholestrogen or diethylstilbestrol (DES) are oxidized to semiquinones and quinones by the peroxidatic activity of cytochrome P-450. The quinones are proposed to be (the) reactive intermediates of estrogen metabolism. (3) The quinones may be reduced to catecholestrogens and DES and redox cycling may ensue. Redox cycling of estrogens has been shown to generate free radicals which may react to form the organic hydroperoxides needed as cofactors for oxidation to quinones. (4) The quinone metabolites of catechol estrogens and of DES bind covalently to DNA in vitro whereas DNA binding in vivo has only been examined for DES. When DES is administered to hamsters, the resulting DES-DNA adduct profile in liver, kidney, or other organs closely matches that of DES quinone-DNA adducts in vitro. In vitro, DES-DNA adducts are chemically unstable and are generated in incubations with organic hydroperoxide as cofactor. It is proposed that the instability of adducts and the lower sensitivity of previous assay methods contributed to the reported failures to detect adducts. Steroid estrogen-DNA adducts in vivo are currently under investigation. (5) Tumors are postulated to arise in cells rapidly proliferating due to the growth stimulus provided by the estrogenic activity of the primary estrogen or of hormonally potent metabolites such as 4-hydroxyestradiol. The covalent modification of DNA in these cells is temporary because of the chemical instability of adducts and will result in altered genetic messages in daughter cells, whereas in non-proliferating cells there may be no lasting genetic damage. The sequence of events described above is a plausible mechanism for tumor initiation by estrogens and is partially substantiated by experimental evidence obtained in vitro and/or in vivo.     

The effects of prenatal tertiary butanol administration in CBA/J and C57BL/6J mice

Pregnant mice of the CBA/J and C57BL/6J strains were given either tertiary butanol (10.5 mmoles/kg, p.o.) or an equivalent volume of tap water twice daily from day 6 through day 18 of gestation. Examination on day 18 revealed significantly more resorptions per litter in the t-butanol-treated animals but no interstrain difference. Tertiary butanol did not significantly affect the body weight of the survivors nor produce significant abnormalities in either strain. Subsequent blood concentration profiles in female C57BL/6J mice indicated that the treatment regimen produced blood levels equivalent to teratogenic ethanol treatment. Mice receiving 3 days of t-butanol treatment did not eliminate the drug more rapidly than control animals, indicating that tolerance was not a factor in the treatment regimen. Since t-butanol shares membrane disordering effects with ethanol but is not metabolized by the same pathway, a role for acetaldehyde or the process of ethanol metabolism is suggested in ethanol teratogenicity.     

Mixing gilts in early pregnancy does not affect embryo survival

There is general acceptance that mixing sows during the first 3 weeks of gestation is detrimental to embryo development and survival. However, there is a paucity of data describing the influence of group housing and remixing during the first 14 days of gestation on pregnancy outcomes. Using 96 purebred maternal (Large White)/terminal (Duroc) line gilts, the current study determined the effects of regrouping, and the timing of regrouping, during the pre-implantation period on embryo mortality. The study was conducted in 2 blocks, with 12 gilts allocated to each of 4 treatments in each block. At 175 days of age, the combination of PG600 and 20 min of daily physical boar contact was used to stimulate puberty, with boar contact resuming 12 days after first detection of oestrus and gilts receiving two artificial inseminations (AIs), 24 h apart, at their second oestrus. After their first AI gilts were allocated to one of four treatment groups (n=12 gilts/treatment). Gilts in one treatment group were housed individually in stalls (STALL). The remaining gilts continued to be housed in their pre-AI groups and were either not remixed (NOMIX), or remixed to form new groups on day 3/4 (RMIXD3/4) or day 8/9 (RMIXD8/9) of gestation (day 0=day of first detection of second oestrus and first insemination). Group-housed gilts were housed in groups of 6, with a space allowance of 2.4 m2/gilt. All gilts were fed once a day (2.2 kg/gilt). Reproductive tracts were collected on day 26.6+/-0.13 of gestation, and the number of corpora lutea (CL) and viable embryos counted. Pregnancy rate was similar across all treatments, averaging 94.5% across the four treatment groups. The number of embryos present on day 26 of gestation was unaffected by housing treatments (P>0.05); gilts in the STALL, NOMIX, RMIXD3/4 and RMIXD8/9 groups possessed 13.2+/-0.67, 12.9+/-0.66, 14.1+/-0.46 and 13.8+/-0.57 embryos, respectively. Similarly, embryo survival rates were 0.91+/-0.04, 0.85+/-0.04, 0.91+/-0.02 and 0.87+/-0.05 for the STALL, NOMIX, RMIXD3.4 and RMIXD8/9 groups, respectively (P>0.05). In conclusion, the current data indicate that individually housing gilts immediately after their first AI does not improve embryo survival. There also appear to be no adverse effects on embryo development or survival when group-housed, mated gilts are remixed during the first 10 days of gestation.     

The effects of branches on prepartum nest building in gilts with access to straw

Sows farrowing in a semi-natural environment terminate nest building 1-7 h prior to parturition after having built a nest for which a variety of materials are used. No nest-building behaviour occurs during parturition and the sows remain lying in the nest throughout most of the farrowing. In contrast, many intensively housed sows are restless during farrowing. To investigate whether gilts housed indoors would use branches for nest building and whether access to branches would affect the termination of nest building and parturient behaviour, we studied gilts housed individually in pens designed to stimulate natural nest building. The control group (n=21) had unlimited access to straw and the experimental group (n=21) had unlimited access to straw and branches. During nest building all the gilts used straw and all the experimental gilts also used branches. In the experimental group the interval from termination of nest building to birth of the first piglet (BFP) was significantly longer than in the control group (132 versus 58 min, P=0.04). In the experimental group, nest-building behaviour was also performed by fewer individuals during the interval from BFP until 2 h after than in the control group (38% versus 71% of the gilts, P=0.03). Gilts that performed nest building during this interval carried out more postural changes (P<0.001) and spent less time in lateral recumbency (P=0.001) than gilts which did not perform nest building. On average, gilts that performed nest building behaviour after BFP (n=26) spent 54% of the first 2 h of parturition in lateral recumbency and carried out 16 postural changes. Gilts that did not perform nest building behaviour during this interval (n=16) spent 85% of the time in lateral recumbency and carried out five postural changes. In 10 gilts that were selected randomly from the experimental group nest building was studied in more detail. In these gilts nest building peaked between 17 and 6 h prepartum. There was no difference in amount of behaviour directed towards straw and amount of behaviour directed towards branches.The results indicate that the termination of nest building in sows is under environmental feedback control. When only straw was provided the nests did not have much of a lasting structure. However, when gilts had access to straw and branches more structured and functional nests could be built. These nests may have been more effective in reducing the motivation for nest building prior to the onset of parturition.     

Effects of in-vivo administration of GnRH on the release of gonadotrophins in the female rabbit

Developing female rabbits were studied weekly from Day 22 of life to Day 100. At all ages GnRH (1.5 micrograms/kg) induced a large increase in LH release 15 min later. By contrast, FSH was significantly increased only on Days 22, 29 and 72 and no significant increase was detected up to 2 h after GnRH administration at other ages. Functional corpora lutea were absent at the start of all treatments as indicated by circulating concentrations of progesterone less than 2 ng/ml. It is concluded that the immature rabbit pituitary is functionally capable of responding to GnRH with an increase in LH secretion, whereas the control of FSH secretion may be regulated by other factors.     

Disentangling prenatal and postnatal maternal genetic effects reveals persistent prenatal effects on offspring growth in mice

Mothers are often the most important determinant of traits expressed by their offspring. These "maternal effects" (MEs) are especially crucial in early development, but can also persist into adulthood. They have been shown to play a role in a diversity of evolutionary and ecological processes, especially when genetically based. Although the importance of MEs is becoming widely appreciated, we know little about their underlying genetic basis. We address the dearth of genetic data by providing a simple approach, using combined genotype information from parents and offspring, to identify "maternal genetic effects" (MGEs) contributing to natural variation in complex traits. Combined with experimental cross-fostering, our approach also allows for the separation of pre- and postnatal MGEs, providing rare insights into prenatal effects. Applying this approach to an experimental mouse population, we identified 13 ME loci affecting body weight, most of which (12/13) exhibited prenatal effects, and nearly half (6/13) exhibiting postnatal effects. MGEs contributed more to variation in body weight than the direct effects of the offsprings' own genotypes until mice reached adulthood, but continued to represent a major component of variation through adulthood. Prenatal effects always contributed more variation than postnatal effects, especially for those effects that persisted into adulthood. These results suggest that MGEs may be an important component of genetic architecture that is generally overlooked in studies focused on direct mapping from genotype to phenotype. Our approach can be used in both experimental and natural populations, providing a widely practicable means of expanding our understanding of MGEs.