Allelic variation in the erythropoietin receptor gene is associated with uterine capacity and litter size in swine

A single nucleotide polymorphism (SNP; C vs. T) that creates an extra GATA-1 site (T allele) in intron 4 of the swine erythropoietin receptor (EPOR) gene was discovered and a genotyping assay for this SNP was developed. A total of 402 gilts from lines selected either at random (control), for ovulation rate (OR) or for uterine capacity (UC) for 11 generations were unilaterally hysterectomized-ovariectomized (UHO) at 160 days of age, mated at approximately 250 days of age and slaughtered at 105 days of pregnancy. Blood samples and spleens were collected from each foetus and the numbers of corpora lutea (CL) and live foetuses, the weights of each foetus and placenta, and each foetal haematocrit were recorded. In addition, intact gilts from the OR line or from a Yorkshire, Landrace, Duroc, crossbred line (BX) were mated and farrowed. At farrowing, the numbers of fully formed and live piglets were recorded for each litter. Genomic DNA was isolated for both the UHO and intact gilts, from foetuses from the UHO gilts that were heterozygous for the EPOR SNP, and from the boars from the BX line and were then used to determine EPOR SNP genotypes. Only CC and CT gilts were observed in the control, OR and UC selected lines. Presence of the EPOR T allele was associated (P < 0.05) with increased UC in these gilts. The number of heterozygous and homozygous foetuses did not differ within UHO litters, or did EPOR genotype influence foetal haematocrit. In intact gilts from the OR line, litter size was significantly associated (P < 0.05) with EPOR SNP genotype. Finally, results from intact gilts of the BX line, in which both the gilt and the boar genotypes were known, allowed an analysis to determine the effect of the gilt and/or the foetal genotype on litter size. This analysis indicated that the predicted foetal genotype (with gilt genotype as covariate) was associated with litter size (an increase of 2.6 +/- 1.0 piglets born alive predicted for homozygous T litters compared with homozygous C litters, P < 0.01) whereas the effect of the gilt genotype (adjusted for foetal genotype) on litter size was not significant. These results indicate that the EPOR SNP is associated with UC and litter size in two distinct populations and could be useful in increasing litter size in swine that are not limited in OR.     

Peripheral plasma prolactin concentrations during oestrous cycles in different types of primitive gilt

Plasma prolactin concentrations were determined by radioimmunoassay during oestrous cycles and around the time of oestrus in different types of primitive gilts: Vietnamese, Zlotnicka and wild-boar X domestic pig hybrids. The animals were bled without stress from an indwelling arterial catheter. The following results were obtained: (1) in all gilts the main prolactin peak was observed at Day 15 or 16 of the oestrous cycle; (2) Vietnamese and hybrid gilts showed a second smaller prolactin surge after (Day 2) or before (Day 17) oestrus; (3) base levels of prolactin during the oestrous cycle were 14.8 +/- 0.93 ng/ml (Vietnamese gilts), 13.2 +/- 1.05 ng/ml (Zlotnicka gilts) and 15.6 +/- 2.01 ng/ml (hybrid gilts). The 15-16-day prolactin peaks reached maximum values of 36.4, 43.4 and 56.5 ng/ml respectively.     

The effect of peri-conception nutrition on embryo quality in the superovulated ewe

Evidence indicates that oocyte/embryo quality in the sheep is affected by nutrient status during the cycle of conception. This study aimed to determine, in the superovulated ewe, if there are stages during the peri-conception period (-18 days to +6 days relative to the day of ovulation [Day 0]) when quality is more likely to be influenced by nutrition. In Experiment 1, ewes were provided with either a 0.5 x maintenance (L), 1.0 x maintenance (M) or 1.5 x maintenance (H) diet (in terms of daily energy requirements) during the peri-conception period. Diet did not affect the mean ovulation rate (range: 15.4+/-1.47 to 16.1+/-1.55) nor the mean number of embryos collected per ewe (range: 10.9+/-2.05 to 12.4+/-1.82) but there was an increase (P<0.05) in the mean number of cells per blastocyst in the L diet (74.7+/-1.45) compared with either the M (66.4+/-1.29) or H (62.0+/-0.84) diets. This increase was due to an increase in the number of trophectoderm (Tr) cells, resulting in a shift (P<0.05) in the Tr:inner cell mass (ICM) cell ratio (range 0.69+/-0.03 to 0.73+/-0.04). In Experiment 2, six diets (HHH, MHH, MHL, MLH, MLL and LLL) were imposed during three 6-day periods commencing 12 days before and continuing until 6 days after ovulation. Although diet had minimal effect on the superovulatory response, both the mean number of cells per blastocyst and the Tr:ICM ratio were increased (P<0.05) when the L diet was provided after Day 0 (diets MHL, MLL and LLL). It is concluded that the ewe is able to respond to acute changes in nutrition imposed immediately after ovulation, resulting in changes in embryo development including cell lineage differentiation. The significance of these findings, in terms of fetal development, embryo-maternal signalling and the nutritional management of the ewe is discussed.     

The effects of PG600 and boar exposure on oestrus detection and potential litter size following mating at either the induced (pubertal) or second oestrus

Within gilt pools, incidences of delayed puberty attainment, failure to exhibit regular oestrous cycles and low first litter size are often high. Boar exposure is an effective method of accelerating puberty; however, the timing of gilt response can vary greatly. Although, PG600 (400 IU of PMSG and 200 IU of hCG; Intervet) can induce a rapid and synchronous ovulatory response, thus providing an alternative to boar contact, the quality of the response is often variable. This study compared the effect of PG600, either alone (NBC) or in conjunction with boar exposure (BC), on puberty attainment and maintenance of oestrous cyclicity. The effects of first mating these gilts at the hormonally induced (pubertal) or second oestrus on ovulation rate and early embryo survival were also studied. Eighty Large White cross terminal (Duroc) line gilts were used in this study. The study was conducted in two blocks, with 10 gilts allocated to each of the four treatments in each block. Gilts were artificially inseminated at the allocated oestrus, with the reproductive tracts collected at 26.5+/-0.29 days after first mating (mean+/-S.E.M.), and the number of corpora lutea and viable embryos recorded. Mean days-to-puberty was significantly reduced (P<0.05) when gilts received both PG600 and boar exposure as opposed to PG600 alone (5.7+/-0.15 versus 6.9+/-0.37 days; P<0.01). The proportion of gilts exhibiting an ovulatory response to PG600 was similar for the BC and NBC treatment groups (0.88 and 0.84); however, the proportion of gilts exhibiting visible signs of oestrus in response to PG600 was significantly higher for the BC compared to the NBC treatment groups (0.81 versus 0.49; P<0.05). Boar contact resulted in a numerical, but not significant, increase in the proportion of gilts exhibited a second oestrus (1.00 versus 0.76). There was no significant effect of boar contact on ovulation rate, embryo number or survival. Although ovulation rate was unaffected by oestrus at mating, embryo number was significantly increased (P<0.05) following mating at the second compared to the first oestrus (11.2+/-0.96 versus 7.8+/-1.17). In conclusion, the current data indicate that the timing of puberty attainment and oestrus detection are significantly improved when PG600 treated gilts receive full boar contact. Further, it is evident that mating gilts at their second as opposed to the hormonally induced oestrus significantly increases embryo number at day 26 post-mating.     

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)     

Effect of dietary intake on pattern of growth of dominant follicles during the oestrous cycle in beef heifers

Friesian x Hereford heifers (n = 19; mean +/- s.e.m. body weight (BW) = 375 +/- 5 kg) were used in a randomized incomplete block design. Heifers were fed 0.7 (n = 7; L), 1.1 (n = 7; M) or 1.8% (n = 5; G) of BW in dry matter (DM)/day for 10 weeks. Ovaries were examined by ultrasound, for one oestrous cycle, from week 5 of treatment. Maximum diameter of dominant follicles was smaller (P less than 0.05) in L (11.8 +/- 0.1 mm) than in M (13.7 +/- 0.2 mm) or G (13.2 +/- 0.3 mm) heifers. Growth rate (mm/day) of dominant follicles during the oestrous cycle was not affected (P greater than 0.05) by dietary intake. Persistence of dominant follicles was shorter (P less than 0.05) in L (9.8 +/- 0.2 days) than in M (11.9 +/- 0.3 days) or G (12.7 +/- 0.4 days) heifers. Three dominant follicles were identified during the oestrous cycle of 5 of 7 L, 3 of 7 M and 1 of 5 G heifers (P less than 0.10); 2 dominant follicles were identified in the remaining heifers (n = 2 of 7, 4 of 7 and 4 of 5, respectively). Length of the luteal phase and luteal-phase concentrations of progesterone were not affected (P greater than 0.05) by treatment. Low dietary intake reduced the diameter and persistence of dominant follicles during the oestrous cycle of beef heifers and tended to increase the proportion of oestrous cycles with 3 dominant follicles.     

Effects of circulating progesterone and insulin on early embryo development in beef heifers

The aims of this study were to determine the effect on early embryo development of feeding a diet formulated to enhance circulating insulin concentrations and secondly to investigate the association between early embryo development and maternal progesterone concentrations in beef heifers. The study was carried out in 32 Simmental x Holstein Friesian heifers 22-25 months of age weighing 506+/-7kg and in condition score 3.1+/-0.1. Animals were fed two diets that were isoenergetic and isonitrogenous, but that would encourage either propionate (diet A) or acetate (diet B) production in the rumen. The rationale was that propionate would induce a greater insulin release in response to feeding. Animals were fed a 50:50 mix of the two diets for 14 days at 0.8x maintenance, with straw provided ad libitum. Animals were then fed one of the experimental diets for 3 weeks prior to synchronisation of oestrus and insemination and for a further 16 days following mating. All heifers were blood sampled daily from oestrus synchronisation and eight animals on each diet underwent daily transrectal real-time ultrasonography to determine the day of ovulation. All heifers were slaughtered at Day 16 after mating. While feeding of diet A (propionic) caused a significant (P<0.05) increase in the plasma insulin to glucagons ratio differences in insulin were not significantly different. This is probably due to the fact that insulin concentrations were quite high as the heifers used in the present study were in good body condition making further increases in insulin difficult to achieve. Diet did not affect size of ovulatory follicle (DIET A: 15.1+/-0.7mm; diet B: 14.6+/-0.7mm), day of ovulation (diet A: 3.5+/-0.2 days; diet B: 3.4+/-0.2 days), mean plasma progesterone concentration (diet A: 4.7+/-0.4ng/ml; diet B: 5.2+/-0.3ng/ml), corpus luteum weight (diet A: 6.0+/-0.2g; diet B: 6.0+/-0.2g) or pregnancy rate (diet A: 81.3%; diet B: 81.3%). However, the proportion of well-elongated (>10cm) embryos on Day 16 was higher in animals fed diet A than in those fed diet B (84.6% versus 38.5%; P<0.05). While progesterone concentration did not differ between pregnant and non-pregnant heifers, progesterone did show an earlier post-ovulatory rise in heifers with well-elongated (>10cm) embryos with levels in these animals significantly higher on Days 4 and 5 than in heifers with small (<10cm) embryos at slaughter. This study demonstrated an enhancement in early embryo development in animals fed a diet generating an increased insulin:glucagon ratio that was not related to circulating maternal progesterone concentrations. However, across diets, enhanced embryo development was associated with elevated plasma progesterone on Days 4 and 5 following mating.     

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.     

Critical periods for foetal mortality in gilts identified by analysing the length distribution of mummified foetuses and frequency of non-fresh stillborn piglets

The objective of this study was to investigate the timing of foetal mortality in gilts of a segregating F2 cross of Large White and Meishan pigs on the basis of the length distribution of mummified foetuses and the frequency of non-fresh stillborn piglets in order to establish whether critical periods for foetal mortality exist. All expelled conceptuses and placentae of 192 farrowing gilts with a normal health status were meticulously investigated to recover all mummified foetuses. The length of each mummified foetus was measured. The predicted number of foetuses present per gilt at the early foetal stage of gestation was calculated as the sum of numbers of mummified foetuses and non-fresh stillborn, fresh stillborn and liveborn piglets. Foetal loss was calculated as the sum of mummified foetuses and non-fresh stillborn piglets. The average foetal mortality rate per gilt was 8.7%. In total 162 mummified foetuses were found (average 0.84 per litter), ranging in length from 0.4 to 33.0 cm. This indicates a range in foetal age at death of approximately 35-100 days. Although mummified foetuses of all lengths within the above mentioned range were found, relatively many had a length of less than 4 cm or of 10-21 cm. The total number of non-fresh stillborn piglets (i.e. late foetal deaths) was 58 (average 0.30 per litter). It can be concluded that foetal mortality occurred in these gilts throughout the period from day 35 to term, with relatively high incidences at the early foetal stage (days 35-40), shortly after mid-pregnancy (days 55-75) and after approximately day 100 of gestation. These three periods coincide with reported periods of change in porcine placental growth.     

Effects of dietary l-arginine supplementation to gilts during early gestation on foetal survival, growth and myofiber formation

The effects of l-arginine on porcine foetal development and myogenesis were determined. Twenty Swiss Large White gilts were randomly allocated to either the control (C) or l-arginine treatment (A). In addition to the standard gestation diet, A-sows received 26 g l-arginine daily from days 14 to 28 of gestation. At day 75 of pregnancy, sows were sacrificed and the number and weight of foetuses were recorded. From each litter, the lightest, heaviest and the ones with an average foetal weight (FtW) were selected. Primary (P), secondary (S) and total myofiber number as well as S/P ratio were determined in the semitendinosus (ST) and rhomboideus (RH) muscles. In A-sows, the number of viable foetuses (13.0 v. 9.3) and total FtW (4925 v. 3729 g) was greater (P ⩽ 0.04) than in C-sows. Compared to C-sow foetuses, the ST of A-sow foetuses had 7% more (17 699 v. 16 477; P = 0.04) P myofibers and the S/P ratio in both muscles was lower (ST = 20.3 v. 21.5; RH = 24.1 v. 27.1; P ⩽ 0.07). Regardless of the maternal diet, the S myofiber number and the S/P ratio in both muscles were greater (P ⩽ 0.01) in foetuses with a high FtW compared to low FtW. These data suggest that l-arginine supplemented to gilts during early gestation enhanced foetal survival and in the ST positively affected the primary phase of myofiber formation.     

Dietary energy source and feeding levels during the rearing period affect ovarian follicular development and oocyte maturation in gilts

Fifty-four Landrace × Yorkshire gilts (59.0 ± 4.2 kg and 147 ± 3 d old) were used to examine the effects of dietary energy source (starch or mixed fat) at high [112.5% of energy requirements recommended by NRC (1998)], normal (100%), and low (87.5%) energy feeding levels on ovarian follicular development and oocyte maturation. Forty-seven estrus gilts were slaughtered at Day 19 after the second estrus; oocytes were recovered from follicles >4 mm in diameter, and matured in vitro for 44 h. Gilts fed high-energy diets had more follicles >4 mm (mean, 25.8 vs. 19.1, P < 0.05) and more oocytes that reached metaphase II (80.3 vs. 64.0%, P < 0.05) than those fed the low-energy diet. Furthermore, gilts fed starch-rich diets had enhanced oocyte nuclear maturation relative to those fed fat-rich diets (75.4 vs. 68.0%, P < 0.05). Compared to the lower-energy feeding groups, high-energy feeding groups had higher (P < 0.05) blood concentrations of postprandial insulin (1562.4 vs. 990.0 ng/4 h), IGF-I (321.2 vs. 256.9 ng/mL), and LH pulses (2.7 vs. 1.4 pulses/6 h). Follicular fluid concentrations of IGF-I (198.5 vs. 143.1 ng/mL) and estradiol (152.6 vs. 124.8 ng/mL) were higher (P < 0.05) in the high-energy group than in the normal group. Compared with gilts fed the high-energy diet supplemented with fat, gilts fed the high-energy diet supplemented with starch had a tendency (P < 0.10) towards increased IGF-I concentration in both blood and follicular fluid, and improved oocyte nuclear maturation during culture in vitro. We inferred that starch-rich, high-energy diets during rearing may improve ovarian follicular development and oocyte maturation in replacement gilts.     

Concentrations of progesterone in the backfat of pigs during the oestrous cycle and after ovariectomy

Small samples of backfat were taken daily during one oestrous cycle and more frequently after ovariectomy from 12 gilts by means of a simple biopsy technique and the levels of progesterone were determined. Compared to the levels of progesterone in peripheral plasma changes in backfat levels during the oestrous cycle were delayed by 1-2 days. Maximal levels with 89.7 +/- 9.2 (mean +/- s.e.m) ng progesterone/100 mg backfat were recorded on Day 15 of the oestrous cycle. It was estimated that, on this day, a total amount of about 36 mg progesterone is stored in the adipose tissue, which is approximately 200 times that present in total blood and corresponds to the daily production of the corpora lutea of the sow on Day 11. Initial half-life of progesterone in backfat after ovariectomy was estimated to be about 34 h compared to an initial half-life of plasma progesterone of about 120 min. The exact calculation of half-lives was, however, confounded by an obvious effect of anaesthesia or surgery on progesterone levels. Changes in backfat or plasma progesterone concentrations were not affected by the fat-to-lean ratio of the gilts. Fat progesterone levels determined in 44 additional pregnant and non-pregnant sows 17 or 20 days after mating indicated that reliable diagnosis of non-pregnant sows was possible on Day 20. It is concluded that the endocrinology of the oestrous cycle in pigs is related to the enormous storage of progesterone in the fat.     

Adenylate cyclase activity of the corpus luteum during the oestrous cycle of the pig

Basal adenylate cyclase values for corpora lutea (CL) removed from cyclic gilts on Days 3, 8, 13 and 18 were 178 +/- 61, 450 +/- 46, 220 +/- 25 and 208 +/- 18 pmol cAMP formed/min/mg protein, respectively. Basal activity was significantly elevated on Day 8 (P less than 0.001). LH-stimulatable adenylate cyclase values for CL from Days 3, 8, 13 and 18 were 242 +/- 83, 598 +/- 84, 261 +/- 27 and 205 +/- 17 pmol cAMP formed/min/mg protein respectively. Serum progesterone concentrations of 12 gilts bled every 2 days through one complete oestrous cycle ranged from 1.1 to 26.9 ng/ml with highest values between Days 8 and 12. The decline in serum progesterone concentrations was coincident with the decrease in basal adenylate cyclase activity. There was no LH-stimulatable adenylate cyclase activity present in the CL at the specific times of the oestrous cycle examined. We conclude that progesterone secretion by the pig CL is apparently dependent on basal activity of adenylate cyclase.     

Prolactin administration during the luteal and follicular phase of the oestrous cycle of gilts

In Exp. I infusions of prolactin (0.5 mg in 2 ml sterile saline) were repeated every 2 h for 36 h on Days 12-13 of the cycle. In Exp. II infusions of prolactin were administered from Days 17 to 19 (60 h) at 2-h intervals. Control gilts were given 2 ml sterile saline at similar intervals during the same period. Basal prolactin concentrations before initiation of infusions ranged from 1.3 +/- 0.1 to 5.6 +/- 2.2 ng/ml in both experiments. By 5 min after a prolactin infusion, mean plasma prolactin concentration ranged from 74.9 +/- 5.8 to 113.0 +/- 9.5 ng/ml, but then declined to approximately equal to 10 ng/ml just before the next infusion of prolactin. Administration of prolactin during the luteal phase of the oestrous cycle of the gilts had no effect on basal levels of progesterone, oestradiol or LH. During the follicular phase there were no differences (P greater than 0.05) between control and prolactin-treated gilt progesterone and LH concentrations, but oestradiol plasma values were decreased (P less than 0.05) on the 2nd and 3rd day of prolactin treatment. Our results would indicate that prolactin does not play a major role in the regulation of the oestrous cycle of the pig.     

The effects of zearalenone on reproduction in swine. I. The relationship between ingested zearalenone dose and anestrus in non-pregnant, sexually mature gilts

Ninety-nine sexually mature, non-pregnant gilts were checked for estrus daily with a mature boar and then allocated at estrus (D O) to receive 2 kg/d of a diet containing 0, 1, 5 or 10 ppm purified zearalenone between D 5 and 20 of the estrous cycle during two seasons of the year (winter and summer). None of the gilts exhibited any visual signs of "hyperestrogenism" and there was no effect of season on interestrous interval (P > 0.05). A significant effect of zearalenone dose on inter-estrous interval was detected (P < 0.001). Gilts receiving 0 or 1 ppm had similar inter-estrous intervals (21.0 +/- 0.3 and 21.5 +/- 0.8 d, respectively) whereas gilts receiving 5 and 10 ppm had extended cycles (29.2 +/- 2.9 and 32.7 +/- 3.3 d, respectively). Plasma progesterone concentrations at D 19 to 21 were higher in gilts with extended cycles (P < 0.001) and corpora lutea (CL) were present at laparotomy. Some 86% of these retained CL underwent spontaneous regression resulting in the onset of estrus within the next 30 d. Fecal zearalenone concentrations rose during ingestion of contaminated diets and declined to pretreatment values within 2 d (1 ppm) to 8 d (10 ppm) of the cessation of treatment. These data show that feeding zearalenone at concentrations of 5 to 10 ppm from D 5 to 20 of the estrous cycle causes luteal maintenance and extended inter-estrous intervals. Spontaneous regression of these CL usually occurs within 30 d after zearalenone is removed from the diet. Fecal zearalenone analysis does not appear to be an effective method for determining prior exposure to zearalenone when carried out more than a few days following the last ingestion of zearalenone.     

Consequences of different dietary energy sources during follicular development on subsequent fertility of cyclic gilts

The objective of the present study was to investigate the effects of dietary-induced insulin enhancement during the late luteal phase on subsequent fertility of gilts. Fifty-two littermate cyclic gilts were subjected to dietary treatments where two energy sources were tested: corn starch (T1) and soybean oil (T2). The experimental diets were supposed to provide similar amounts of dietary energy, but from different sources. Gilts were fed ad libitum, starting day 8 of the estrous cycle, until the next standing heat. Blood sampling was performed in a subgroup of 20 gilts on days 14 and 21 of the cycle for analyses of glucose and insulin, and after ovulation detection until 18 h after ovulation for progesterone. All gilts were slaughtered on day 28 of pregnancy and the reproductive tracts recovered for further analysis. T1 gilts showed higher postprandial insulin peak on days 14 and 21 and lower glucose levels 4 h after feeding on day 14 (P<0.05), however, there were no treatment effects on plasma progesterone concentrations. Dietary energy sources did not affect average daily feed intake, body weight and backfat on day 28 of pregnancy. Estrous cycle length, estrus duration and time of ovulation were not affected by previous nutritional treatments either. T1 gilts showed higher ovulation rates, number of embryos, embryo weight and placental weight (P<0.05). There were no treatment effects on pregnancy rate, embryo survival rate and volume of amniotic fluid. A positive correlation between progesterone concentration 18 h after ovulation and ovulation rate was observed (r=0.75; P<0.01). These results suggest that it is possible to manipulate dietary insulin response in cyclic gilts and, thus, improve reproductive efficiency when feeding starch as the main energy source during the late luteal and follicular phases of the cycle.     

Litter-size-dependent intrauterine growth restriction in sheep

Regulation of foetal development in sheep depends on interactions between the intrinsic capacity of the foetus for growth and the maternal environment. Lambs born in multi-foetus litters have relatively small placentae with fewer cotelydons, and lower birth weights. Litter-size-dependent intrauterine growth restriction (IUGR) is evident at mid gestation when metabolic needs of the conceptus are moderate, and overnutrition of ewes with multiple foetuses does not promote growth of their foetuses to the size of singletons. Those observations suggest that placental and conceptus growth in multi-foetus pregnancies is reprogrammed at mid gestation by an as yet undefined mechanism to attenuate foetal growth. This may protect the foetus from severe nutritional insult during late gestation, when its daily growth rate is at a maximum. In that way, lambs born in large litters with relatively lower birth weights may not experience the long-term physiological insults that can be observed in small lambs born to undernourished ewes.     

Embryo yield and quality following dietary supplementation of beef heifers with n-3 polyunsaturated fatty acids (PUFA)

The objective of this study was to examine the effects of dietary n-3 polyunsaturated fatty acid (n-3 PUFA) supplementation on embryo yield and quality in heifers. Animals were individually offered barley straw and concentrate diets supplemented with either palmitic acid (C16:0; CON) or a partially rumen protected n-3 PUFA-enriched supplement. Following oestrous cycle synchronisation, superovulation was induced using FSH. Blood samples were collected for the measurement of fatty acids, metabolites, insulin and IGF-1. On day 7 post-insemination the number of ovulations was estimated and embryos recovered non-surgically and quality graded. At embryo recovery 50ml of the uterine flushing was collected from each horn for fatty acid analysis. Grade 1 embryos were isolated, snap frozen in liquid nitrogen and stored at -80 degrees C. mRNA expression for six genes, LIF, BAX, Cx43 and E-CAD associated with embryo development, and PPAR-alpha and -delta, associated with lipid metabolism was analysed. The n-3 PUFA supplementation increased plasma n-3 PUFA concentration (P<0.05) and reduced n-6:n-3 PUFA ratio (P<0.05). Uterine concentration of the n-3 PUFA, eicosapentaenoic acid was increased (P<0.05) and the concentration of arachidonic acid decreased (P<0.05) following n-3 PUFA supplementation. While CON increased triglyceride concentrations, diet did not affect the other plasma metabolites, insulin or IGF-1 (P>0.05). Similarly, there was no effect of diet on superovulation rate, embryo recovery rate, embryo quality (P>0.05) or mRNA expression of the genes examined (P>0.05).     

Effect of intrauterine infusion of Escherichia coli on hormonal patterns in gilts during the oestrous cycle

The aim of this study was to determine the effect of intrauterine Escherichia coli infusion on the patterns of plasma LH, prolactin, progesterone, androstenedione, testosterone, oestrone, oestradiol-17beta, cortisol and 13,14-dihydro-15-keto-prostaglandin F2alpha (PGFM) in gilts during the oestrous cycle. On day 4 of the oestrous cycle (day 0), 25 mL of saline or 25 mL of Escherichia coli suspension, containing 10(7) colony forming units x mL(-1), was infused once into the each uterine horn in group I or II respectively. The control gilts developed a new oestrous cycle at the expected time but not bacteria-treated. Endometritis and vaginal discharge developed in all gilts after Escherichia coli infusion. The administration of Escherichia coli resulted in a reduction of plasma levels of LH, prolactin, oestrone and oestradiol-17beta (P < 0.05-0.001), mainly on days 15-18 after treatment (expected perioestrous period). During this time, the plasma androstenedione level was elevated (P < 0.05-0.001) after bacteria infusion. In the gilts receiving bacteria, progesterone concentration decreased from day 8 after treatment and was low until the end of the study (P < 0.05-0.001). On days 8-12 after bacteria administration, the level of PGFM was higher (P < 0.001) than that found in the control group. These results suggest that the developing inflammatory process of the endometrium in gilts following Escherichia coli infusion significantly affects the pituitary-ovarian axis function as well as prostaglandin production leading to anoestrus.     

Embryo survival and recipient pregnancy rates after transfer of fresh or vitrified,in vivo or in vitro produced ovine blastocysts

The aim of this study was to assess the effect of production system and of cryopreservation of ovine embryos on their viability when transferred to recipients. The experimental design was an unbalanced 2 x 2 factorial design of two embryo production systems (in vivo versus in vitro) and two embryo preservation conditions prior to transfer (transferred fresh versus transferred after vitrification/warming). For the production of blastocysts in vivo, crossbred donor ewes (n=30) were synchronised using a 13-day intravaginal progestagen pessary. Ewes received 1500 IU equine chorionic gonadotropin (eCG) 2 days before pessary withdrawal, and were mated 2 days after pessary withdrawal and embryos were recovered surgically (6 days after mating). Blastocysts were produced in vitro (IVP) using standard techniques. Recipients (n=95) were synchronised using a progestagen pessary and received 500 IU eCG at pessary removal and were randomly assigned to receive (two per recipient) in vivo fresh (n=10), in vivo vitrified (n=10), in vitro fresh (n=35) or in vitro vitrified (n=40) blastocysts. Recipients were slaughtered at day 42 of gestation and foetuses recovered. Pregnancy and embryo survival rates were recorded and analysed using CATMOD procedures. Foetal weights and crown-rump lengths were recorded and analysed using generalised linear model (GLM) procedures. There were no statistically significant interactions between the effects of embryo production system and preservation status at transfer on pregnancy rate and embryo survival. The pregnancy rate following transfer of fresh IVP blastocysts was lower (P<0.07) than that of in vivo embryos (54.3% versus 90.0%, respectively). Vitrification resulted in a decrease in pregnancy rate, the effect being more pronounced in the case of IVP embryos (54.3-5.0%, P<0.001) compared with in vivo embryos (90.0-50.0%), although the absolute change was similar (49.3% versus 40.0%). Transfer of fresh IVP blastocysts resulted in a higher proportion of single (78.9% versus 33.3%) and lower proportion of twin (21.1% versus 66.7%) pregnancies than those produced in vivo. This was reflected in a significant difference in embryo survival rate (fresh: 32.8% versus 75.0%, P<0.01; vitrified: 2.5% versus 35.0%, P<0.001, for IVP and in vivo blastocysts, respectively). Similarly, all pregnancies resulting from the transfer of vitrified/warmed IVP blastocysts were single pregnancies, while 40% of those from vitrified/warmed in vivo blastocysts were twin pregnancies; this was reflected in an embryo survival rate of 35.0% versus 75.0%, respectively. There was a significant effect (P=0.0184) of litter size on foetal weight but not on foetal length (P=0.3304). Foetuses derived from the fresh transfer of IVP blastocysts were heavier (6.4+/-0.2g versus 5.8+/-0.2g, respectively, P<0.05) and longer (5.2+/-0.1cm versus 4.8+/-0.1cm, respectively, P<0.01) than those derived from fresh in vivo blastocysts. There was no difference in these parameters as a consequence of vitrification of IVP embryos. However, in vivo blastocysts subjected to vitrification resulted in heavier (6.6+/-0.3g versus 5.8+/-0.2g, respectively, P=0.055) and longer (5.2+/-0.1cm versus 4.8+/-0.1cm, respectively, P<0.05) foetuses than their counterparts transferred fresh.