Evaluation of systems for collection of porcine zygotes for DNA microinjection and transfer

Crossbred gilts and sows (n=116) were used for the collection of 1-cell zygotes for DNA microinjection and transfer. Retrospectively, estrus synchronization and superovulation schemes were evaluated to assess practicality for zygote collection. Four synchronization and superovulation procedures were used: 1) sows were observed for natural estrous behavior; 1000 IU human chorionic gonadotrophin (hCG) was administered at the onset of estrus (NAT); 2) cyclic gilts were synchronized with 17.6 mg altrenogest (ALT)/day for 15 to 19 days followed by superovulation with 1500 IU pregnant mares serum gonadotropin (PMSG) and 500 IU hCG (LALT); 3) gilts between 11 and 16 days of the estrous cycle received 17.6 mg ALT for 5 to 9 days and PMSG and hCG were used to induce superovulation (SALT); and 4) precocious ovulation was induced in prepubertal gilts with PMSG and hCG (PRE). A total of 505 DNA microinjected embryos transferred into 17 recipients produced 7 litters and 50 piglets, of which 8 were transgenic. The NAT sows had less (P < 0.05) ovarian activity than gilts synchronized and superovulated by all the other procedures. Synchronization treatments with PMSG did not differ (P > 0.05) in the number of corpora hemorrhagica or unovulated follicles, but SALT and PRE treaments had higher ovulation rates than LALT (24.7 +/- 2.9, 24.3 +/- 1.8 vs 11.6 +/- 2.7 ovulations; X +/- SEM). The SALT and PRE treatments yielded 12.3 +/- 2.6 and 17.7 +/- 1.7 zygotes. Successful transgenesis was accomplished with SALT and PRE procedures for estrus synchronization and superovulation.     

Successful nonsurgical deep uterine embryo transfer in pigs

At present, it is possible to transfer pig embryos directly into the uterine body of sows by nonsurgical procedures. The aim of this study was to develop a procedure for nonsurgical embryo transfer (ET) into the upper part of one uterine horn in gilts and sows. In experiment 1, 29 gilts and 43 sows were used. Intrauterine insertions took place for each female at days 4-6 of the estrous cycle (D0 = onset of estrus). An artificial insemination (AI) spirette was inserted into the cervix to assist with the guidance of a modified flexible catheter originally developed for deep intrauterine insemination in pigs. The flexible catheter length inserted anterior to the inserted AI spirette was 43.0 +/- 1.7 cm. The time required to complete the procedure was affected by the type of female (P < 0.001) and by the difficulties encountered for inserting the catheter (P < 0.001). However, when no or minor difficulties were encountered during the insertion of the catheter (in approximately 70 and 80% of gilts and sows, respectively), the time required to complete the procedure did not differ between gilts (2.5 +/- 0.1 min) and sows (2.3 +/- 0.1 min). In experiment 2, 24 to 31 fresh morulae and/or blastocysts were transferred to each of 24 recipients. Seventeen animals (70.8%) farrowed an average of 6.9 +/- 0.7 piglets, of which 0.6 +/- 0.3 piglets were born dead. In conclusion, the procedure described in this study offers new possibilities to transfer embryos nonsurgically to the uterine horn of pigs.     

Effects of parity and synchronization of donors by an oral progestogen on embryo transfer in swine

The aim of this study was to establish whether the quantity or quality of embryos collected is affected by 1) the reproductive status of donors (nulliparous gilts vs parous sows); 2) pretreatment of donors with Oxolven. Embryos were collected from gilts (n=38) and from weaned sows (n=35). Approximately half of each group (gilts and sows) had been subjected to oral treatment with the progestogenic 19-nortestosterone derivative Oxolven for 14 to 21 days. After induction of estrus with pregnant mare serum gonadotropin (PMSG) and human chorionic gonadotropin (hCG) and following artificial insemination, the embryos were recovered surgically. The ovulation rate of donors averaged 21.6, with no significant difference among groups. Most of the ova and embryos collected were at the 4-cell stage (53%). There was a high incidence of uncleaved ova (28%) in both groups of animals. This was particularly so in gilts, both the Oxolven-treated and controls (35 and 32%). The incidence of uncleaved ova was lower, however, in Oxolven-treated (29%) and control sows (14%). Embryos at the 4- to 8-cell stage were transferred to synchronous recipient gilts. Transfer results, expressed by the survival of transferred embryos, were not significantly affected by the progestogen treatment (30% for Oxolven treatment vs 34% for the controls) or by the reproductive status of the donors (33% for gilts vs 28% for sows).     

Effect of exogenous gonadotropins on the weaning-to-estrus interval in sows

The efficacy of PG 600 (400 IU PMSG and 200 IU hCG) for accelerating the onset of estrus was determined for sows weaned during the summer. Yorkshire sows (average parity = 4.6), nursing 8.6 +/- 0.2 pigs (mean +/- SEM) were weaned after 27.7 +/- 0.4 d of lactation and were treated intramuscularly with either PG 600 (n = 35) or with 0.9% saline (n = 35). Sows were checked for estrus once daily in the presence of a mature boar. Treatment with PG 600 increased (P < 0.05) the percentage of sows in estrus within 7 d after weaning (97.1 vs 82.9%). Relative to controls, sows given PG 600 expressed estrus sooner (3.8 +/- 0.1 d vs 4.5 +/- 0.1 d; P < 0.01). Sows exhibiting estrus within 7 d after treatments were artificially inseminated 0 and 24 h after first exhibiting estrus. The percentage of inseminated sows that farrowed tended to be higher (P < 0.07) for control than for PG 600-treated sows (96.6 vs 82.3%). The number of pigs born live was similar (P > 0.1) for sows treated with PG 600 and with saline, and was 12.7 +/- 0.6 and 11.7 +/- 0.7, respectively. Pigs farrowed by saline-treated sows, however, tended to be heavier (P < 0.09) than pigs farrowed by sows treated with PG 600 (1.49 +/- 0.06 kg vs 1.34 +/- 0.06 kg). In summary, PG 600 accelerated the onset of estrus in sows weaned during the summer. Sows mated during the induced estrus, however, tended to have a lower farrowing rate and farrowed lighter pigs than control sows inseminated during a natural estrus occurring within 7 d after weaning.     

Development of porcine embryos from one- and two-cell stages to blastocysts in culture medium supplemented with porcine oviductal fluid

Oviductal fluid (OVF) was harvested chronically from 5 sows beginning on Day 1 of the estrous cycle (Day 0 of estrous cycle = day of detected estrus) and used for embryo culture (Day 3 OVF only). Two experiments were conducted to investigate in vitro development of 1-cell and 2-cell porcine embryos in a modified Kreb's Ringer bicarbonate medium (culture medium, CM), early luteal phase OVF or CM supplemented with OVF (CM-OVF, 25% OVF v/v in CM) with or without transfer to fresh CM. In Experiment 1, 1-cell and 2-cell embryos were harvested from sows (n = 7) approximately 44 h after detected estrus. In Experiment 2, 1-cell embryos were collected from 5 sows treated with altrenogest and gonadotropins, approximately 50 h after injection of human chorionic gonadotropin. The volume of OVF (ml) declined progressively throughout the 4 days of collection (24 h, 8.44 +/- 0.28; 48 h, 6.88 +/- 1.78; 72 h, 4.96 +/- 0.35; 96 h, 4.64 +/- 0.25 after onset of estrus; p less than .01). In both experiments, development to blastocyst stage was lowest among embryos cultured in OVF and highest among those cultured in CM-OVF (Experiment 1: CM, 27.3; OVF, 10; CM-OVF, 63.6; Experiment 2: CM, 26.7; OVF, 0; CM-OVF, 82.4; % blastocyst formation).(ABSTRACT TRUNCATED AT 250 WORDS)     

Control of ovulation with a GnRH analog in gilts and sows

Methods for the control of ovulation with GnRH or the GnRH analog D-Phe6 -LHRH (GnRH-A), were evaluated in gilts and sows as the last step in development of a fixed-time Al protocol. This involved 3 field trials using 2,744 gilts (10 units) and 71,628 sows (33 units). In Trial 1, the GnRH-A (75 microg) was given subsequent to treatment with altrenogest for cycle control or eCG for the stimulation of uniform follicle development in gilts. The release of LH was followed by ovulations which commenced within 36.4 +/- 3.3 hr and were terminated at 39.0 +/- 2.8 hr after administration of GnRH-A. This degree of synchronization of ovulations enabled the use of fixed-time AI. Consequently, subsequent to pretreatment with altrenogest and eCG, in 10 production units 1,285 gilts received 50 microg GnRH-A and 1,459 gilts 500 IU hCG serving as positive controls (Trial 2); all the gilts were inseminated 24 and 42 hr after treatment. Pregnancy rate and piglet index (n of piglets per 100 first inseminations) following GnRH-A vs hCG were 78.8% and 779 vs 74.4% and 728, respectively (P < 0.05). In field trials with first litter gilts and multiparous sows (33 units holding from 250 to 6,000 sows), 1,000 IU eCG was used for estrus control after weaning and 25 microg or 50 microg GnRH-A were given 55 to 58 hours after eCG (n = 19,954 and 20,701) (Trial 3). Sows treated during the same time period with 300 microg GnRH plus 300 IU. hCG (n = 30,973) served as positive controls; all sows were inseminated 24 and 42 hours after treatment. Pregnancy rates for 50 microg GnRH-A, 25 microg GnRH-A and 300 microg GnRH plus 300 IU hCG were 83.0%, 81.7% and 80.7%, and the piglet indices 913, 899 and 880, respectively (P < 0.05). Unit size and parity had significant effects on fertility and productivity. In all studies, results with 50 microg GnRH-A were superior. In year-long studies, highest levels of fertility in response to these treatments were seen from December to May.     

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.     

Nonsurgical embryo collection from sows with surgically shortened uteri

To allow for the nonsurgical collection of swine embryos, the uteri of sows (n=7) were surgically shortened. A section of each uterine horn was resected to facilitate a transcervical flushing procedure. All sows with a shortened uterus exhibited natural estrus at least once after the operation. Four to six days after insemination, embryos were collected with a two-way Foley catheter. Embryos were collected (n=55, 6.3+/-6.0: x +/-SD ) from every treated sow. Although treated sows often did not exhibit estrus beyond 1 to 9 natural estruses, those sows (n=27) with persistent corpora lutea (CL) over a 4 to 5 wk period were given prostaglandin F(2alpha) (PGF(2alpha)) and they returned to estrus in 5.2+/-1.1 d: x +/-SD .     

The effects of administration of gonadotrophins and estrogens on prenatal survival in gilts

In gilts ovulation occurs over a 4 to 8-hour period, with 70% of the ova being shed over a relatively short span of time. These oocytes supposedly give rise to more developed embryos at Days 10 to 12 which advance the uterine environment and reduce survival rates of less developed embryos because of an asynchronous environment. The aim of this experiment was to reduce embryo mortality by influencing the duration and pattern of ovulation. Crossbred gilts (n = 98) were bred at their first observed estrus after being exposed to boars at 200 days of age. Estrus detection was carried out daily at 0000, 0800 and 1600 hours. All gilts were artifically inseminated with fresh semen, with a minimum of 2.7 billion spermatozoa, at both 16 and 32 hours after detection of estrus. Gilts were randomly assigned to one of the following treatments at detection of estrus: 1) 500 IU (2ml) chorionic gonadotrophin (hCG) injected intravenously at the onset of estrus (n = 22); 2) 16 mug (4 ml) gonadotrophin releasing hormone (GnRH) injected intravenously at the onset of estrus (n = 25); 3) 11.5 mug estrogen added to the semen at the time of AI (n = 25); 4) control, untreated gilts (n = 26). All gilts were slaughtered at Day 30 of gestation (Day 0 = day of detected estrus). The mean (+/-SEM) number of ovulations in pregnant gilts per treatment was 13.0 +/- 0.52, 12.6+/-0.51, 13.6+/-0.54 and 13.3+/-0.52, while the mean (+/-SEM) number of normal embryos per treatment was 10.3+/-0.67, 10.5+/-0.66, 10.3 +/- 0.69 and 10.5 +/- 0.67 for hCG, GnRH, estrogen and control groups, respectively, for an embryonic survival rate of 80 +/- 4.2%, 83 +/- 4.1%, 74 +/- 4.3% and 79+/-4.2% in pregnant gilts. If nonpregnant gilts are included, the embryonic survival rate for treatments 1 to 4 was 76+/-7.0%, 73+/-6.5%, 60+/-6.5%, and 64+/-6.4%, respectively. There was no significant difference between treatments for any of these variables. There was no evidence that administration of hCG, or GnRH at the onset of estrus, or the addition of estrogen to semen improved embryonic survival in gilts by Day 30 in this experiment.     

Influence of SLA haplotype on ovulation rate and litter size in miniature pigs

Systemic blood was collected from and surgery performed on sows of 3 strains of miniature swine bred for specific SLA (swine MHC) haplotypes (a, c and d) from Day 2 to Day 6 after mating (first day of mating = Day 0). Ovulation rate was determined by counting corpora lutea and embryos were flushed from the uterus. Progesterone, oestradiol-17 beta and oestrone were quantitated in blood plasma and uterine flushings by RIA. SLAd/d females had a higher ovulation rate than SLAa/a or SLAc/c females (11.50 +/- 0.87 vs 9.11 +/- 0.68 and 8.17 +/- 0.83, respectively; P less than 0.01). Oestrone was higher than oestradiol-17 beta in systemic plasma (56.5 +/- 6.4 vs 33.0 +/- 4.7 pg/ml, P less than 0.01) while oestradiol-17 beta was higher than oestrone in uterine flushings (19.8 +/- 1.4 vs 14.9 +/- 1.5 pg/horn, P less than 0.10). Systemic progesterone concentration was correlated with day after mating (r = 0.93, P less than 0.01). There was no effect of haplotype on any of the hormone concentrations measured. Litter size was analysed from 99 matings amongst SLAa/a, SLAa/c, SLAa/d, SLAd/c and SLAd/d sires and dams. Litter size from -/d and d/d sows or from d/d boars were larger (P less than 0.05) than for all other matings. Although ovulation rate was higher in SLAd/d sows, the significant effect of sire SLA genotype on litter size suggests an additional effect of the d haplotype on embryonic survival.     

In vitro development of embryos from Sinclair miniature pigs: a preliminary report

The goal of this project was to identify conditions that result in development from the zygote or the 2-cell stage Sinclair miniature pig embryos to the blastocyst stage. Four media were selected, 2 that have been shown to result in in vitro development in domestic pigs (Hepes buffered Tyrode's medium and Whitten's medium), 1 that is compatible with similar development in the cow (CR-1), and 1 that is compatible with development in the mouse (CZB). One- and two-cell stage embryos from Sinclair miniature pigs were flushed from oviducts in Hepes buffered Tyrode's medium, allocated to 1 of the 4 media and cultured for 120 h. At the end of the culture period, embryos were morphologically scored and nuclei were counted. Morphology scores were lowest for Hepes buffered Tyrode's medium but were not different for Whitten's medium, CZB or CR-1. The highest (P < 0.07) number of nuclei was present in the oocytes cultured in Whitten's medium (21.3), with CR-1 (15.7) and CZB (16.5) not differing significantly. Similar to the morphology scores, Hepes buffered Tyrode's medium resulted in the lowest number nuclei (5.5). In a parallel experiment, domestic pig embryos were cultured in Hepes buffered Tyrode's medium versus Whitten's medium. The domestic pig embryos, while also developing better in Whitten's Medium, developed better in the Hepes buffered Tyrode's medium than did the embryos from Sinclair pigs. Thus, the Sinclair pig embryo develops best if placed in Whitten's Medium.     

Synchronization and superovulation of mature cycling gilts for the collection of pronuclear stage embryos

An efficient protocol was developed to synchronize and superovulate mature pigs for the collection of pronuclear stage embryos suitable for DNA microinjection. A timed and coordinated regimen of Lutalyse, PG600 and Chorulon along with daily checking for estrus allowed synchronization of groups of gilts having estrous cycles at regular intervals. Pigs 10-16 days after the beginning of standing estrus have been successfully synchronized into estrus using this protocol. A standard dose of each drug was used independent of size or age of the animal. One protocol averaged 38.9 ovulations and 31.1 one-cell embryos recovered per animal.     

The effect of modified eros centre, outdoor raising or conventional group housing on breeding gilts and its effects on reproductive performance over four parities

The present study was conducted in a large Croatian "built up unit". The objective of the study was to determine if an indoor modified eros centre (MEC) compared to indoor or outdoor group housing of gilts, influenced the onset of puberty of gilts and the reproductive performance of the evaluated females (n = 783) over four parities. The gilts were from the same nucleus herd. Gilts of same age (140-150 days of age), body condition (body condition score of 3-4) and similar genetics (four-way cross females), during the same season (January to April 1999), were randomly divided at arrival into three groups and treated as follows:MEC gilts (n = 279): These were placed into indoor MEC pens in groups of 8-10. The gilts had continuous fenceline contact to boars (one boar to two groups of gilts, boars were changed daily) and to shortly weaned oestrous sows. Gilts were regrouped and dislocated at 10-day intervals. Outdoor gilts (n = 263): These were kept in groups of 8-10 on a large pasture (80-100 m2 per group). The animals had fenceline contact to mature boar for 5-10 min daily. Control indoor gilts (n = 241): These were housed indoors in large pens in groups of 8-10. The animals had fenceline contact to mature boars for 5-10 min daily. Each outdoor group had an insulated hut with straw bedding. All gilts were fed ad libitum with the same commercial diet. Housing gilts in MEC resulted in earlier (P < 0.001) onset of estrus (MEC: 174.8 +/- 2.4 days, indoor group housing: 207.6 +/- 4.1 days, outdoor group housing: 187.4 +/- 2.1 days) and lower (P < 0.001) farrowing rate to first service (MEC: 70.97%, indoor group housing: 89.73%, outdoor group housing: 89.62%). Farrowing rate of regularly returning MEC gilts to second service was 95.00%. First total-born litter size, first liveborn litter size, first wean-to-estrus interval (WEI), percent of sows bred after first weaning, second total-born litter size, second liveborn litter size, average third and fourth total-born and liveborn litter size, number of sows having four litters, number of litters per sow, total number of pigs per sow, total number of liveborn pigs per sow showed no significant differences between the groups. More (P < 0.05) sows were culled in outdoor group. Compared to MEC and outdoor housing, indoor housed sows suffered higher (P < 0.05) percentage of anoestrus.     

Piglets born after non-surgical deep intrauterine transfer of vitrified blastocysts in gilts

The aims of this study were: (1) to evaluate the effect of the number of previous estrus of recipient gilts on effectiveness of intrauterine insertion of a flexible catheter designed for non-surgical deep intrauterine catheterization during diestrus in pigs; and (2) to determine the farrowing rate and the litter size after non-surgical deep intrauterine embryo transfer (ET) of porcine blastocysts vitrified by the open pulled straw (OPS) method. In experiment 1, 27 large white hyperprolific gilts (LWh) with 2-6 previous estrus were used. Intrauterine insertions of the flexible catheter were carried out at day 5.5-6 of the estrous cycle (D0=onset of estrus). During insertions, no or only moderate reactions were observed in 88.9% of gilts and was not related (P >0.05) to the number of estrus prior to the insertion periods. The number of the estrus had a significant effect (P <0.05) on the difficulties found during the procedure. In the 100% of gilts with two estrus (N=6) it was not possible to insert the flexible catheter through the cervix. In gilts with three or more estrus, it was possible to pass the cervix and to progress along a uterine horn in 80.9% of the cases. In 86.7% of the gilts, the tip of the flexible catheter achieved the second or third quarter of the uterine horn. In experiment 2, following non-surgical deep intrauterine transfer of 20 vitrified/warmed blastocysts, 9 Meishan recipients (42.9%) farrowed an average of 5.4 +/- 0.8 piglets (range 3-9) of which 0.6 +/- 0.3 piglets (range 0-2) were born dead. In conclusion, this study shows that it is possible to obtain birth of piglets following non-surgical deep intrauterine embryo transfer (ET) of vitrified/warmed blastocysts. Non-surgical deep intrauterine ET and OPS vitrification methods are promising procedures to be used together for the introduction of new genetic material in a farm.     

Development of in vivo-matured porcine oocytes following intracytoplasmic sperm injection

The objective of this study was to assess the development of porcine ova fertilized by intracytoplasmic sperm injection (ICSI). Allyl trenbolone (Regumate) was used to synchronize estrus in 13 postpuberal gilts. Gilts were superovulated with pregnant mare serum gonadotropin and hCG. Ova were aspirated from 5- to 8-mm follicles at 36 h after hCG. Cumulus cells were removed by blunt dissection and pipetting in Beltsville embryo culture medium (BECM) supplemented with 0.1% hyaluronidase. Sperm were washed and resuspended in BECM + 8% polyvinylpyrrolidone. Ova (n = 237) that exhibited a polar body were centrifuged at 15 000 x g for 6 min and injected with a single spermatozoon. One hundred fifty-four ova were cultured in NCSU-23 medium in a 5% CO(2) in air environment for 168 h. Ova were fixed in acetic acid/ethanol and stained with 1% orcein. Sixty-nine ICSI ova were cultured for 24 h and transferred (mean = 23) to three recipients. Eighty-one ova (69%) that survived ICSI cleaved within 48 h. Thirty-eight percent (31/81) of these ova became blastocysts (mean +/- SEM = 24.7 +/- 1.1 cells). One recipient gave birth to three pigs. These results demonstrate that porcine embryos derived from ICSI can develop into live pigs.     

Ovarian response, embryo recovery and results of embryo transfer in a Hungarian native pig breed

The objective of the study was to use embryo transfer (ET) for propagation of the Swallow Belly Mangalica population. Mangalica is a native Hungarian pig breed adapted to extreme climate and housing conditions and distinguished for excellent meat and fat quality. However, due to their weak reproductive characteristics and relatively high fat proportion, Mangalica pigs have been replaced by modern breeds. Now, there is an increased interest again to safeguard the properties of this breed. We conducted two experiments. First, we used a total of 18 puberal Mangalica gilts to determine an optimal superovulatory treatment. Following estrus synchronization with Regumate, we injected gilts with either 750, 1000 or 1250 IU PMSG, followed by 750 IU hCG 80 h later. We scanned ovaries endoscopically 3 days after hCG administration. The application of 1000 and 1250 IU PMSG resulted in a higher rate of ovulation compared to 750 IU (24.2 +/- 3.6 and 21.0 +/- 2.3 vs. 13.7 +/- 2.7 P<0.05). The number of follicular cysts increased after administration of 1250 IU PMSG compared to 750 and 1000 IU (2.0 +/- 1.3 vs. 0.3 +/- 0.7 and 0.2 +/- 0.3, P<0.05). Thus, we chose 1000 IU PMSG for further stimulation of Mangalica gilts. In the second experiment, we induced superovulation in 10 Mangalica donor gilts by 1000 IU PMSG and 750 IU hCG. Gilts were fixed-time inseminated, and then five days later embryo collection was carried out surgically (n=6) or endoscopically (n=4). Out of the 187 ova recovered, 92.5% were at the morula/blastocyst stage. The embryo recovery rate was higher following surgical flushing than following endoscopy (91.5 +/- 4.4% vs. 71.4 +/- 12.7%, P<0.05). Altogether 143 embryos were transferred surgically or endoscopically into 8 Landrace recipients. Surgical and endoscopic transfer of Mangalica embryos into Landrace gilts resulted in pregnancies in 3 and 2 gilts, respectively; thus the overall farrowing rate was 62.5%. The birth of 59 Mangalica piglets from 5 embryo recipients equals an average litter size of 11.8 +/- 1.3, which is two times larger than usual in this breed. Therefore, we concluded that an appropriate inter-breed ET program is a suitable tool to propagate the endangered Mangalica breed.     

In vitro development of embryos from superovulated gilts treated with the progesterone agonist, altrenogest (Regu-Mate) or the prostaglandin analogue, cloprostenol (Planate)

This study was conducted to compare the in vitro development of embryos from superovulated postpubertal gilts synchronized with progesterone agonist altrenogest (REG, Regu-Mate) and those from superovulated prepubertal gilts synchronized with prostaglandin analogue cloprostenol (PLA, Planate). Ten postpubertal gilts that had exhibited estrus at least once were fed 20 mg/d of REG from Day 0 (the first day of treatment, may have been any day of the estrous cycle) to Day 17. The gilts received intramuscularly (im) 1500 IU of equine chorionic gonadotropin (eCG) on the afternoon of Day 17, followed by 1000 IU of human chorionic gonadotropin (hCG) 84 h later. Eight prepubertal gilts received intramuscularly one dose of a combination of 400 IU of eCG and 200 IU of hCG (PG 600) on Day 0 (the first day of treatment), followed by 750 IU of hCG on Day 3. From Day 16 to Day 19, the prepubertal gilts received 350 mg/d of PLA, followed by 1500 IU of eCG on the afternoon of Day 19, then 1000 IU of hCG 84 h later. Gilts were checked for estrus with an intact boar. At estrus, all gilts were artificially inseminated and/or mated twice at 12-h intervals. Then 50 to 54 h after the hCG injection, a mid-ventral laparotomy was performed on each gilt. Corpora albicans (CA) and corpora hemorrhagica (CH) were counted, and oviducts were flushed in situ. The embryos recovered (1- to 2-cell) were cultured in modified Whitten's medium at 38.5 degrees C under an atmosphere of 5% CO2 in air for 144 h. The number of CA per gilt did not differ between the postpubertal and prepubertal gilts (11.9 vs 7.9, respectively; P > 0.05). However, the number of CH per gilt (27.5 vs 18.1, P = 0.05) and the number of embryos per gilt (26.2 vs 15.3, P < 0.05) were higher in postpubertal gilts than in prepubertal gilts. Furthermore, after 144 h of in vitro culture, the percentage of embryos cleaving to the >-16-cell (morula + blastocysts) or > or =32-cell (blastocysts) was greater (P < 0.05) in prepubertal gilts than in postpubertal gilts (85.2 vs 68.5, 55.7 vs 44.2, respectively). The total numbers of embryos examined were 122 and 260 in prepubertal and postpubertal gilts, respectively. These results show that postpubertal gilts treated with REG produced a higher number of embryos. However, better embryo development was noted with zygotes from prepubertal gilts primed with exogenous gonadotrophin, followed by synchronization with prostaglandin before induction of superovulation and insemination.     

Common swine leucocyte antigen (SLA) haplotypes in NIH and Sinclair miniature swine have similar effects on the expression of an inherited melanoma

Mixed lymphocyte culture and serological typing of NIH and Sinclair miniature swine indicate that the two herds share a common SLA haplotype. NIH haplotype a (International Haplotype H10) appears identical to Sinclair haplotype B, which has significant effects on the penetrance of Sinclair swine cutaneous malignant melanoma (SSCM). Offspring of crosses between melanoma-bearing Sinclair swine homozygous for the B haplotype and non-melanoma NIH aa swine have tumour incidence identical to Sinclair melanoma BB× Sinclair non-melanoma BB offspring. Our results provide further support for the involvement of the swine leucocyte antigen (SLA) complex in the inheritance of SSCM, and identify a new source of non-tumour animals that have all of the genes for SSCM except those at the tumour-initiator locus.     

The influence of insulin administration after weaning the first litter on ovulation rate and embryo survival in sows

Primiparous crossbred sows (n = 43), lactating for an average of 21.1 +/- 0.1 d and weaning 8.7 +/- 0.1 pigs, were used to evaluate the influence of insulin on ovulation rate and embryo survival. The sows were maintained on 2.3 kg/head/d of a 14% protein gestation diet during pregnancy, fed ad libitum during lactation, given 2.7 kg/head/d from weaning until re-breeding and fed 2.3 kg/head/d after mating. Beginning the day after weaning (Day 0) sows were treated with 0.4 IU/kg body weight (BW) insulin (n = 21) or were administered an equivalent volume of saline (n = 22) for 4 d. Beginning on Day 3 and continuing until Day 14 after weaning, the sows were checked for estrus twice daily and were artificially inseminated using pooled semen from 2 fertile boars. At slaughter (days 30 to 40 of gestation), ovaries and uteri were collected, and the ovulation rate, embryo number and viability, and uterine weight and length were evaluated and recorded. Use of insulin decreased the average interval from weaning to estrus compared with saline by increasing percentage in estrus by Day 14 after weaning (5.0 +/- 0.57 vs 6.9 +/- 0.56 d, respectively; P < 0.03). Ovulation rate, number of embryos, embryo survival, and average uterine length and weight were not influenced by insulin treatment. Overall, insulin affected reproductive efficiency in primiparous sows by increasing the percentage of sows in estrus.     

Time of the preovulatory LH surge in the gilt and sow relative to the onset of behavioral estrus

Two experiments were conducted to study the time of occurrence of the preovulatory LH surge in pigs. Sampling every ten minutes in six cycling gilts before and after onset of standing estrus revealed the preovulatory surge began from 8 hr before to 12 hr after the lordosis reflex was elicited. Three of six gilts initiated the preovulatory LH release coincident with the onset of estrus. Data from 28 postpartum sows, with samples drawn every six hours commencing with the onset of estrus, indicated maximum LH levels were present at the first observance of estrus. Six of the 28 sows had an LH peak 18-24 hr after the onset of estrus.