Regression of induced corpora lutea by human chorionic gonadotropin in prepuberal gilts

The effect of daily injections of human chorionic gonadotropin (HCG) on luteal maintenance in hysterectomized prepuberal gilts induced to ovulate and in hysterectomized mature gilts was studied. Twenty-four pre-puberal gilts, 120 to 130 d of age, were induced to ovulate with 1,000 IU pregnant mare serum gonadotropin followed 72 h later with 500 IU HCG. Nine of the 24 prepuberal gilts (bred controls) were artificially inseminated on d 0 (d 0 = d after HCG). Mature gilts that had displayed one or more estrous cycles of 17 to 22 d were used (d 0 = onset of estrus). All gilts, except the bred controls, were totally hysterectomized on d 6 to 9 and their corpora lutea (CL) marked with charcoal. From d 10 through 29, eight prepuberal and 10 mature hysterectomized gilts received daily injections of 500 IU HCG in saline while seven prepuberal and eight mature hysterectomized gilts received daily injections of saline vehicle. Jugular blood samples were quantitated by radioimmunoassay for estrogen and 13,14-dihydro-15-keto prostaglandin F2 alpha (PGFM), a metabolite of prostaglandin F2 alpha. One bred control gilt was pregnant on d 30, indicating that the prepuberal gilts used in the experiment were prepuberal. All mature gilts and six of seven prepuberal gilts that received saline had maintained CL to d 30. Eight of 10 mature gilts that received HCG had maintained CL to d 30, while only two of eight (P less than .05) prepuberal gilts that received HCG maintained CL to d 30. All gilts receiving HCG had numerous follicles and accessory luteal structures.(ABSTRACT TRUNCATED AT 250 WORDS)     

Precocious estrus and reproductive ability induced by PG 600 in prepuberal gilts

A total of 29 SPF Large White prepuberal gilts (mean age 152 days at treatment) were examined for estrous and ovulatory responses after PG 600 treatment. After treatment, 85.2% of the gilts showed standing estrus within 6 days. Whereas the treatment-to-estrus interval and duration were 3.7 and 1.9 days respectively. As ovulation occurred on Day 5 to 6, appropriate timing of artificial insemination would be about 4 days after treatment. Fertility of gilts revealed to be excellent, giving rise to a high percentage of normal embryos, 85.3%. Meanwhile, development and growth of fetuses were mostly normal. Other reproductive performances recorded were: mean litter size 6.8; mean birth weight 1.26 kg; weaning-to-return estrus interval 5 to 8 days. In conclusion, PG 600 was found to be useful in inducing fertile estrus in prepuberal gilts, a result which will be of interest for commercial pig farms.     

Comparison of induced corpora lutea from prepuberal gilts and spontaneous corpora lutea from mature gilts: in vitro progesterone production

Prepuberal (P) gilts were induced to ovulate with pregnant mare serum gonadotropin followed 72 h later by human chorionic gonadotropin (hCG). Three P gilts and three mature (M) gilts each were ovariectomized on d 10, 14, 18, 22 and 26 (d 0 = day of hCG for P gilts and onset of estrus for M gilts). Gilts ovariectomized on d 14, 18, 22 and 26 were hysterectomized on d 6 to ensure maintenance of the corpora lutea (CL). Two to five grams of minced luteal tissue were dispersed using collagenase and hyaluronidase in HEPES buffered salt solution supplemented with glucose and bovine serum albumin. Dispersed cells were rinsed in Dulbecco's Modified Eagle Medium (DMEM), counted (ratio of large to total number of luteal cells determined) and then incubated for 1 h in DMEM. With aliquots standardized to 2.5 X 10(4) viable, large cells (greater than 25 micron diameter) were incubated in 1 ml DMEM for 2 h in the presence of either 10, 50, 100 or 1,000 ng luteinizing hormone (LH); .1, 1, 10 or 100 ng hCG; 10, 100 or 1,000 ng norepinephrine (NE) or either .75, or 1.5 mM dibutyrl cyclic adenosine monophosphate (dbcAMP). Progesterone (P4) in the medium was quantified by radioimmunoassay. Basal P4 production (no P4 stimulator added to the medium) on d 10, 14, 18, 22 and 26 for P gilts was 246 +/- 9, 66 +/- 4, 64 +/- 6, 41 +/- 3 and 69 +/- 6 ng/ml medium, respectively, and for M gilts was 281 +/- 12, 128 +/- 8, 53 +/- 4, 82 +/- 6, 101 +/- 5 ng/ml medium, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)     

Study of follicular development and heredity of morphological types of ovaries in prepuberal gilts

Two different morphological types of ovaries ("grape" and "other" types) previously described by microscopic examination were studied on 107 live gilts. The ovarian type and distribution of macroscopic follicles during the prepuberal period were determined at 140, 160 and 180 d of age by laparoscopic examination. The incidence of grape-type (GT) ovaries was higher (P less than .05) at 140 d of age (50.5%) than at 160 d (38.3%) or 180 d (35.5%). At maturity (d 19 of the first postweaning estrus), 37.7% of the ovaries were classified as GT. The proportion of gilts that maintained their initial type (at 140 d of age) at 160 d, 180 d and at maturity was higher (P less than .05) for the gilts having the other-type (OT) ovaries (49% and 25% for OT and GT, respectively). The mean number of small follicles (1 to 3 mm in diameter) was higher (P less than .05) for the OT ovaries at 140 d (71.5 vs 18.0), at 160 d (79.3 vs 26.2) and at 180 d (77.0 vs 32.8). The number of follicles of 4 to 5 mm was higher (P less than .05) for GT ovaries at 140 d (16.4 vs 4.8), at 160 d (20.9 vs 4.8) and at 180 d of age (21.4 vs 5.7). The large follicles (greater than or equal to 6 mm) were more numerous for the GT at 140 d (7.0 vs .3), at 160 d (5.7 vs .5) and at 180 d (4.4 vs .6). No statistical difference was observed at maturity in the distribution of follicles between the two types.(ABSTRACT TRUNCATED AT 250 WORDS)     

Comparison of induced corpora lutea from prepuberal gilts and spontaneous corpora lutea from mature gilts: hydroxysteroid dehydrogenase activity

The activity of hydroxysteroid dehydrogenases was histochemically quantified in corpora lutea (CL) from prepuberal gilts induced to ovulate and mature gilts. Prepuberal (P) gilts, 120 to 130 d of age were induced to ovulate with 1,500 IU pregnant mare serum gonadotropin (PMSG) followed 72 h later by 500 IU human chorionic gonadotropin (hCG). Three P gilts and three mature (M) gilts each were ovariectomized on d 10, 14, 18, 22 and 26 (d 0 = day of hCG for P gilts and onset of estrus for M gilts). Gilts ovariectomized on d 14, 18, 22 and 26 were hysterectomized on d 6 to ensure luteal maintenance. At the time of ovariectomy, CL were frozen in liquid nitrogen and then stored at -80 C until analysis. Cryostat sections (12 microns) were histochemically analyzed for delta 5-3 beta-hydroxysteroid dehydrogenase (3 beta OHSD), 17 alpha-hydroxysteroid dehydrogenase (17 alpha OHSD) and 20 alpha-hydroxysteroid dehydrogenase (20 alpha OHSD). The intensity of staining (greater enzyme activity resulted in darker staining) was quantified using a Zeiss SF microscope integrated with a Zonax photometer, which measured the percentage of light transmitted through a given area (22,500 microns 2) of the tissue section. Data were subjected to analysis of variance using the general linear models procedure of Statistical Analysis System (SAS). The 3 beta OHSD activity did not change over days, but the mean activity (throughout all days) in the P gilts (32.6 +/- 1.8) tended (P less than .08) to be elevated above that of M gilts (27.9 +/- 1.7).(ABSTRACT TRUNCATED AT 250 WORDS)     

Luteinizing hormone receptor number and affinity in corpora lutea from prepuberal gilts induced to ovulate and spontaneous corpora lutea of mature gilts

This study was designed to determine if luteal cell receptors for luteinizing hormone/human chorionic gonadotropin (LH/hCG) contribute to the previously demonstrated abnormal function of induced corpora lutea (CL) in gilts. Twenty-five prepuberal (P) gilts, induced to ovulate with 1,500 IU pregnant mare serum gonadotropin followed 72 h later with 500 IU hCG (d 0 = day of hCG), and 22 mature (M) gilts that had displayed two or more estrous cycles were ovariectomized (OVX) on d 10, 14, 18, 22 or 26 after the onset of estrus. All gilts except those OVX on d 10 were hysterectomized between d 6 and 9 to ensure luteal maintenance. The CL were stored at -196 degrees C until determination of LH/hCG receptor number and dissociation constant (KD) by saturation analysis. Receptor number was greater for M than for P gilts on d 14 (P less than .07) and d 18 (P less than .01). The KD was greater in M than in P gilts on d 14 (P less than .01) and d 18 (P less than .0001). The LH/hCG receptor number and KD of P gilts remained the same throughout the days studied. The LH/hCG receptor number (fmol/mg protein) of M gilts was elevated on d 10, 14, and 18 (50.8, 50.4 and 51.4, respectively) and decreased on d 22 (26.5) and d 26 (25.4) to values similar to those of P gilts. In M gilts, KD increased on d 14, remained high on d 18 and decreased on d 22. We suggest that abnormal function of induced CL in P gilts may be due to an elevated LH receptor number.     

Effects of gonadotropin treatment on ovarian follicle growth and granulosal cell aromatase activity in prepuberal gilts

This experiment was conducted to compare the ability of USDA porcine FSH-B-1 (pFSH), USDA porcine LH-B-1 (pLH), and pregnant mare's serum gonadotropin (PMSG) to grow large follicles and induce granulosal cell aromatase activity in prepuberal gilts. Twenty-four gilts (164 d old) received one of four treatments by i.m. injection: 1) saline once, n = 8; 2) pFSH (8 micrograms/kg BW, nine times at 8-h intervals), n = 5; 3) pLH (2 micrograms/kg BW, nine times at 8-h intervals), n = 6; or 4) PMSG (15 IU/kg BW, once), n = 5. At slaughter, 72 h after the first injection, the ovaries to saline-treated gilts contained an average of 104 surface antral follicles 1 to 3 mm in diameter. compared to treatment with saline, pFSH increased (P less than .05) the number of follicles 46%, whereas pLH or PMSG decreased (P less than .05) the number by 70 and 84%, respectively. Compared with saline, treatment with PMSG or pLH induced growth of large follicles (7 to 9 mm) (10.8 and 4.8 follicles/gilt, respectively), increased plasma estrogen, increased granulosal cell aromatase activity, and decreased plasma FSH by 51 and 69%; treatment with pFSH had no significant effect on these traits. Results indicate that injected pFSH did not cause growth of large follicles or induce granulosal cell aromatase activity in prepuberal gilts. In contrast, LH initiated growth and increased granulosal cell aromatase activity in a small number of follicles and accelerated atresia among the remaining follicles.     

Effects of exogenous insulin and body condition on metabolic hormones and gonadotropin-induced follicular development in prepuberal gilts

To determine influences of insulin and body condition on follicular growth, prepuberal gilts (n = 16) treated with pregnant mare's serum gonadotropin (PMSG) were used in a 2 X 2 factorial experiment with main effects of insulin (0 or .4 IU/kg every 12 h beginning at 1800 on the day before PMSG) and backfat depth (moderate, 25 +/- .8; high, 32 +/- .7 mm; P less than .0001). Body weights were similar. Blood sampling was at 6-h intervals for analyses of LH, FSH, growth hormone (GH), glucagon, cortisol, insulin, insulin-like growth factor-I (IGF-I), plasma urea nitrogen (PUN), nonesterified fatty acids (NEFA), testosterone, estradiol-17 beta, and progesterone. Ovaries were removed 75 h after PMSG treatment, and visible small (less than or equal to 3 mm), medium (4 to 6 mm), large (greater than or equal to 7 mm), and macroscopically atretic follicles were counted. Administration of insulin increased IGF-I in fluid of medium follicles (108.8 vs 60.7 ng/ml; SEM = 13.3; P less than .05). Neither insulin nor fatness affected hCG binding by granulosa cells (12.5 +/- 1.6 ng/10(6) cells) or numbers of large (16.7 +/- 2.6) and medium (10.4 +/- 2.3) follicles. However, insulin increased the number of small follicles (58.9 vs 29.9; SEM = 9.7; P less than .05) and reduced the number of atretic follicles (3.8 vs 11.3; SEM = 1.1; P less than .05). The predominant effect of insulin on reducing number of atretic follicles was in the small size class (.6 vs 6.9; SEM = .6, P less than .01). Follicular fluid estradiol and progesterone were not affected by treatments; however, testosterone concentrations in large follicles were lower in gilts with higher backfat (32.5 vs 59.9 ng/ml; SEM = 4.0; P less than .05). Systemic LH, FSH, glucagon, cortisol, PUN, NEFA, estradiol, and testosterone were not affected by insulin or level of feeding. However, GH was lower in gilts that had higher backfat (overall average of 3.2 vs 2.8 ng/ml; SEM = .1; P less than .05). Insulin reduced atresia and altered intrafollicular IGF-I independently of body condition and without sustained effects on other hormones.     

In vitro fertilisation of in vivo matured porcine oocytes obtained from prepuberal gilts at different time intervals after hCG injection

The goal of the present study was to find out the best interval after hCG injection in PMSG primed prepuberal gilts for retrieval of in vivo matured oocytes for in vitro fertilisation (IVF). Altogether 66 gilts were superovulated with 1500 IU PMSG and 500 IU hCG 72 h later. Ovum pick up was performed endoscopically 24, 28, 32 or 36 h after hCG and a total of 869 cumulus-oocyte-complexes (COCs) were aspirated from 1400 follicles. COCs were tested for quality, and an aliquot was immediately fixed and stained to determine meiotic configuration. The remaining COCs were fertilised in vitro using frozen-thawed epididymal semen. Quality and developmental stage of embryos were tested after IVF, and the number of nuclei was counted. At 24 to 32 h after hCG only few oocytes have entered the second meiotic cycle (18 to 25% vs. 58% at 36 h, p < 0.05). The overall cleavage rate was significantly influenced by insufficient maturation rate at the early collection times (14% at 24 h vs. 49% at 36 h). Additionally, when oocytes were collected 24 to 32 h vs. 36 h the cleavage rate based on mature oocytes was lower (26 vs. 62%, p < 0.05). Once embryonic development has been initiated, the further in vitro development to blastocyst stages did not differ between groups. However, the number of cells was lower at collection times 24 to 32 h as compared to 36 h after hCG (12 to 15 cells vs. 22 cells, p < 0.05). The results indicate that the time of COC collection affects the in vitro developmental competence up to the blastocyst stage and should not be performed earlier than 36 h after hCG treatment.     

Induction of fertile estrus in prepuberal gilts by treatment with a combination of pregnant mare's serum gonadotropin and human chorionic gonadotropin

Ten trials involving 678 presumed prepuberal gilts (5.5 to 7.5 mo old) were conducted in North Carolina, Illinois and Missouri to evaluate the reproductive performance of gilts given a combination of 400 IU of pregnant mare's serum gonadotropin and 200 IU of human chorionic gonadotropin (P. G. 600). Gilts that were presumed to be prepuberal received P. G. 600 or no treatment (control) on the day of movement from finishing facilities to pens for breeding. Detection of estrus, with the aid of mature boars, was conducted daily for 28 d; gilts in estrus were mated naturally. Treatment with P. G. 600 increased the percentage in estrus within 7 (57.5 vs 40.9%) or 28 d (72.9 vs 59.5%); average interval to estrus was reduced (P less than .05) from 10.4 to 7.5 d. Farrowing rate (78.5 +/- 3.1%), number of pigs born alive (8.6 +/- .2) or dead (.26 +/- .06) and number of pigs weaned (8.0 +/- .2) were unaffected by treatment. Gilts that were heavier than the median for each farm were in heat sooner and more were detected in heat, but no other reproductive traits differed between heavy and light gilts. Overall, the results reveal that P. G. 600 was useful for induction of fertile estrus in prepuberal gilts.     

Follicular development during the prepuberal period of different morphological types of ovaries in Hampshire and Yorkshire gilts fed two planes of nutrition

Changes in follicular development of the ovary at 105, 140 and 175 d of age were observed on 48 prepuberal Yorkshire and Hampshire gilts raised on two planes of nutrition. The follicles from the left ovary were counted and measured by histological techniques. Both the nonatretic and the atretic (more than four pyknotic bodies) antral follicles were classified into six categories according to size, and each category was expressed as percentage of the total number. The proportion of nonatretic follicles belonging to the first category (.19 to .36 mm in diameter) was higher in Hampshire than Yorkshire gilts at d 105 (65.4 vs 47.6%; P less than .05), but not at d 140 and 175. The proportion of atretic follicles to the total number observed was higher in Yorkshire at d 175 than at d 140 and 105 (38.6 vs 21.9 vs 11.6%, respectively; P less than .05), whereas in Hampshire, significant differences were observed only between d 140 and 105 (25.2 vs 3.9%, respectively). The low-plane regimen reduced the percentage of nonatretic follicles of the third (.63 to 1.12 mm) and fourth (1.13 to 2.00 mm) categories in Yorkshire gilts from 14.4 and 7.2% to 9.7 and 4.0%, respectively (P less than .05). According to grouping, size and number of antral follicles, three morphological types of ovaries could be distinguished: honey-comb, grape-like and an intermediate class. Reanalyzing the data accounting for types resulted in radical reduction in the residual variation, hence many significant differences between the two breeds could be identified. At 140 and 175 d of age, Hampshires with honey-comb- and grape-like ovaries had greater numbers of medium and large follicles than Yorkshires (P less than .001). It is concluded that the slower follicular development observed at 105 d of age in Hampshire, as compared with Yorkshire gilts, allows accumulation of greater numbers of antral follicles at 140 and 175 d for both morphological types of ovaries.     

Insulin sensitivity during pregnancy, lactation, and postweaning in primiparous gilts

The objectives were to examine changes in the insulin response during pregnancy, lactation, and postweaning in an experiment involving 10 primiparous Landrace x Large White gilts. Gilts were catheterized at 50 d of pregnancy, and tests were conducted at approximately 59 d of pregnancy (midpregnancy; MP), 106 d of pregnancy (end of pregnancy; EP), 17 d of lactation (L), and 9 d after weaning (PW), respectively. Changes in plasma glucose, insulin, and NEFA concentrations were studied after 3 different tests: ingestion of 1.3 kg of feed (meal test); a glucose tolerance test; and 2 euglycemic, hyperinsulinemic clamp tests, in which 20 and 55 ng of insulin x kg of BW(-1) x min(-1) were infused during 150 min. Fasting concentrations of plasma glucose were less during L than during the other stages (P < 0.001). Concentrations of glucose and insulin increased after ingestion of the meal and decreased thereafter. Plasma insulin returned to basal concentrations at all stages, whereas glucose reached basal concentrations before the end of the meal at the PW test only. Postprandial concentrations of plasma glucose and area under the curve for insulin were greater during L than at the other stages (P < 0.05); both tended to be greater during EP than during MP or after weaning. Concentrations of NEFA were greater during L than at other stages before as well as after a meal (P < 0.001). Glucose half-life was greatest during L, least during MP and PW, and intermediate during EP. Compared with other stages, insulin secretion during the tolerance tests seemed to be delayed during L and, to a lesser extent, at EP. Irrespective of insulin dose, glucose infusion rates during the clamps did not differ between MP and PW, and were greater than during EP and L (P < 0.001). Plasma concentrations of NEFA decreased less rapidly during L than during the other stages. Gilts from EP developed a state of insulin resistance that was further accentuated during L. Changes in insulin responsiveness at MP, EP, and L may be an adaptation that allows gilts to acclimate to the increasing demand of glucose by the growing conceptus and the even greater demands of lactation.     

Effects of increased ambient temperature and supplemental altrenogest before pregnancy establishment in gilts

Heat stress (HS) mitigation strategies are critically needed to combat the substantial economic effects on animal agriculture. The manifestations of seasonal infertility include delayed puberty onset, reduced conception rates, decreased litter size, and increased wean to estrus interval. To assess the effects of HS during early gestation and evaluate the benefit of supplemental altrenogest (ALT) as a mitigation strategy, 30 crossbred postpubertal gilts (157 ± 11 kg body weight) were subjected to estrous synchronization via 14 d oral administration of ALT. Artificial insemination during estrus was performed, and gilts were then placed into one of four treatment groups: HS (35 ± 1 °C for 12 h/31.60 ± 1 °C for 12 h) with (HSALT, n = 7) or without (HSCON, n = 7) 15 mg/d ALT supplementation or thermal neutral (TN; 20 ± 1 °C) conditions with (TNALT, n = 8) or without (TNCON, n = 8) 15 mg/d ALT supplementation until 12 d post-estrus (dpe). Administrating ALT occurred at 0600 hours from 3 to 12 dpe, and rectal temperatures (T_R) and respiration rates (RR) were recorded. Blood was collected via jugular venipuncture on 0, 4, 8, and 12 dpe. Gilts were euthanized humanely at 12 dpe followed by the collection of ovarian tissue, and uterine flushing for conceptus collection. In HS compared with TN gilts, RR and T_R were increased (P < 0.01) but unaffected by ALT supplementation. Feed intake was reduced (P < 0.01) by HS but unaltered by the ALT treatment. Corpora lutea (CL) weight was reduced (P < 0.01) in HSCON gilts when compared with TNCON and HSALT gilts despite progesterone concentrations in serum and luteal tissue not being affected by treatment (P ≥ 0.10). CL diameter was reduced (P ≤ 0.05) in HSALT gilts compared with other treatments. Interleukin-1β (IL1B) uterine flush concentration was not affected (P > 0.20) by environment or ALT supplementation, although moderate (P = 0.06) interaction between environment and ALT existed, as IL1B concentration in TNALT was increased (P = 0.03) compared with TNCON gilts. While environment did not affect conceptus development (P = 0.90), ALT supplementation advanced conceptus elongation (P < 0.01). Collectively, these data demonstrate that HS may affect luteal development before pregnancy establishment, and ALT increases conceptus elongation by 12 dpe.