The effects of zearalenone on reproduction in swine. II. The effect on puberty attainment and postweaning rebreeding performance

Eighty-five prepuberal, crossbred gilts received, ad libitum, a diet containing 0 or 10 ppm purified zearalenone for 30 d beginning at 145 to 193 d of age. At the end of this period all gilts were placed on the control diet and exposed daily to a mature boar for 60 d. Within 3 to 5 d of zearalenone ingestion, gilts showed marked vulval swelling and reddening, which continued for the 30-d feeding period. Thereafter symptoms slowly subsided. Zearalenone treated gilts showed first estrus significantly later than controls (P < 0.05), but the proportion of gilts showing estrus within 60 d of boar exposure was similar (P > 0.05). The length of the first estrous cycle was not affected by the ingestion of zearalenone before puberty (P > 0.05). In a second study, 65 multiparous, crossbred sows were full-fed twice daily a ration containing 0 or 10 ppm of purified zearalenone beginning 14 d before weaning. Postweaning, all sows were fed the control diet, were checked for estrus daily, and inseminated at the first postweaning estrus. Neither sows nor gilts from their litters exhibited signs of hyperestrogenism during treatment. Weaning to estrus interval was significantly extended in zearalenone treated sows (P < 0.05), but all other variables of fertility assessed were similar. These data suggest that zearalenone ingestion before puberty delays the stimulation of puberty associated with boar exposure, but does not affect subsequent cyclicity if zearalenone is removed from the ration. Similarly, zearalenone ingestion during lactation delays the return to estrus after weaning, but does not affect subsequent fertility when removed from the ration at weaning.     

Impact of Mated Female Nonproductive Days in Breeding Herd after Porcine Epidemic Diarrhea Virus Outbreak

Porcine epidemic diarrhea virus (PEDV) is an important pathogen that has a significant economic impact on the swine industry by imposing a high rate of mortality in suckling piglets. However, limited information on the productivity values of gilts and sows infected with PEDV is available. Here, we evaluate the productivity index in gilts and sows during the 1-year period before (19 January 2013 to 18 January 2014) and after (19 January 2014 to 18 January 2015) a PEDV outbreak from a 2000-sow breeding herd in Taiwan. The farrowing rate (FR), return rate (RR), total pigs born per litter (TB), pigs born alive per litter (BA), weaning pigs per litter (WPL), pre-weaning mortality, percentage of sows mated by 7 days after weaning, weaning to first service interval (WFSI), mated female nonproductive days (NPDs), replacement rate of sows and sow culling rate were compared using productive records. The FR (-9.6%), RR (+9.8%), TB (-1.6), BA (-1.1), WPL (-1.1), sows mated by 7 days after weaning (-6.9%), WFSI (+0.8 days), NPDs (+6.9 days) and sow culling rate (+7.2%) were significantly different between the 1-year pre-PEDV outbreak period and the post-PEDV outbreak period. Impacts of the PEDV infection on the reproductive performance were more severe in pregnant gilts than in sows. In conclusion, these findings indicate that the outbreak of PEDV caused an increase in the rate of NPDs in breeding herds.     

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.     

Comparative studies on cell lines established from normal and radiation-exposed miniature swine.

Cloned cell lines were established from two swine with radiation-induced myeloproliferative disorders, including one cell culture from an animal with myelogenous leukemia and one from an animal with myeloid metaplasia. A third cloned cell line with similar morphology was established from pooled normal fetal swine cornea to compare the growth characteristics of cells from normal and irradiated swine. All three cell lines grew as foci of aggregated cells and were able to form macroscopic colonies in semisolid agar medium. The lack of normal mechanisms of contact inhibition and the observed aneuploidy indicated that these cells were morphologically transformed. Further, the cloned cells caused tumors in nude mice, clearly indicating that these cells were also malignantly transformed. A major difference between these cell lines was that type C viruses were observed only in the cells derived from swine with myeloproliferative disorders.     

Comparative studies on cell lines established from normal and radiation-exposed miniature swine

Summary Cloned cell lines were established from two swine with radiation-induced myeloproliferative disorders, including one cell culture from an animal with myelogenous leukemia and one from an animal with myeloid metaplasia. A third cloned cell line with similar morphology was established from pooled normal fetal swine cornea to compare the growth characteristics of cells from normal and irradiated swine. All three cell lines grew as foci of aggregated cells and were able to form macroscopic colonies in semisolid agar medium. The lack of normal mechanisms of contact inhibition and the observed aneuploidy indicated that these cells were morphologically transformed. Further, the cloned cells caused tumors in nude mice, clearly indicating that these cells were also malignantly transformed. A major difference between these cell lines was that type C viruses were observed only in the cells derived from swine with myeloproliferative disorders. Work supported by the U.S. Department of Energy under Contract EY-76-C-06-1830.     

Progeny of somatic cell nuclear transfer (SCNT) pig clones are phenotypically similar to non-cloned pigs

Systematic studies of cloned animals generated from adult somatic cell nuclei are critical in assessing the utility of somatic cell cloning in various applications, including the safety of food products from cloned animals and their offspring. Previously, we compared somatic cell derived cloned pigs with naturally bred control pigs on a series of physiological and genetic parameters. We have extended our studies to the F1 progeny of these clones to see whether these phenotypic differences are transmitted to the next generation. There were no differences in the average litter size between litters from cloned gilts and naturally bred controls (7.78 +/- 2.6 and 7.40 +/- 3.0, respectively; mean +/- SD) or in the degree of litter size variation (coefficients of variation of 33.4% and 40.5% for litters of clones and controls, respectively). Similarly there were no statistical differences between sex ratios of cloned litters (51-49%, M:F) and control litters (59-41%, M:F). Blood profiles between cloned pigs, control pigs, and their progeny were compared at two time points (i.e., 15 and 27 weeks) to quantify the effect of cloning on various blood parameters and their transmission to the next generation. Although the range of values for all traits overlapped between different classes, the variability between all the traits in F1 progeny of clones and the control pigs was similar at 15 and 27 weeks, with one exception. Combined, our data and previous results in mice strongly support the hypothesis that offspring of clones are similar to offspring of naturally bred animals, and as such there should not be any increased risks associated with consumption of products from these animals.     

Comparison of milk produced by cows cloned by nuclear transfer with milk from non-cloned cows

Cloning technologies, including embryo splitting and nuclear transfer, were introduced into dairy cattle breeding in the early 1980s. With the recent worldwide attention on the cloning of sheep ("Dolly") and cows ("Gene"), the potential food safety concerns for food products derived from cloned animals needs to be addressed. There has been no study of the composition of milk produced by cloned cows. In this preliminary study, we evaluated the composition of milk from 15 lactating non-embryonic cell cloned cows and six non-cloned lactating cows over a single season. The cloned cows came from five unique genetic lines and three distinct breeds. Milk samples were analyzed for total solids, fat, fatty acid profile, lactose, protein and compared to non-cloned and literature values. Gross chemical composition of milk from cloned cows was similar to that of the non-cloned cows and literature values. Our results lead us to conclude that there are no obvious differences in milk composition produced from cloned cows compared to non-cloned cows.     

Epidemiological Studies of Piglet Diarrhoea in Intensively Managed Danish Sow Herds

The study comprised 70,796 litters in 104 sow herds, observed from 1976 through 1982. Weaning age decreased from approx. 42 days to approx. 30 days during the observation period. Diseases and symptoms were recorded together with production parameters (feeding, barn construction, economic returns etc.). The mean incidence rate of pre-weaning diarrhoea was 6.8 % of litters, with considerable inter-herd differences (incidence rates from 0 to approx. 50%). There was a slight increase in incidence during the autumn (August through October). Incidence rates increased with litter size, with a peak incidence in litters of 11–13 piglets, and decreased with increasing parity of the sow. There was a positive association between occurrence of arthritis and pre-weaning diarrhoea in the litters, and litters from sows with post parturient disease (MMA complex) had 1.8 times higher risk of getting diarrhoea than litters from healthy sows. No important differences among breeds were found. Small herds (< 200 farrowings per year) had higher incidence rates than large herds (400–499 farrowings per year). Herds with a gilt proportion above 30 % had an incidence rate of 12.3 %, i. e. nearly twice as high as the overall mean (6.8 %). There was a trend towards a higher incidence rate in litters kept in traditional pens (i. e. large pens with solid floor and loose sows) than in intensive pens (i. e. small pens with slatted flooring and tethered sows). The overall pre-weaning mortality rate was 16.2 % of pigs born, half of which was due to stillbirths (6.3 %) and overlaid piglets (2.2 %). In litters with pre-weaning diarrhoea, the mortality rate was 19 %, compared to 13 % in litters without occurrence of diarrhoea. This difference accounts for an excess loss of 0.6 piglets from birth to weaning in diarrhoeic vs. non-diarrhoeic litters. Piglets from litters with pre-weaning diarrhoea had reduced weight gain. Thus, on the average, they were 2.2 days older at 25 kg bodyweight and weighed 0.4 kg less at 30 days than piglets from non-diarrhoeic litters. Also, litters with pre-weaning diarrhoea had a significantly increased risk of post-weaning diarrhoea. The present information forms a basis for defining acceptable disease thresholds in suckling litters in intensively managed herds.     

Effects of dietary enrichment with a marine oil-based n-3 LCPUFA supplement in sows with predicted birth weight phenotypes on birth litter quality and growth performance to weaning

The effects of a marine oil-based n-3 long-chain polyunsaturated fatty acid (mLCPUFA) supplement fed to the sow from weaning, through the rebreeding period, during gestation and until end of lactation on litter characteristics from birth until weaning were studied in sows with known litter birth weight phenotypes. It was hypothesized that low birth weight (LBW) litters would benefit more from mLCPUFA supplementation than high birth weight litters. A total of 163 sows (mean parity=4.9±0.9) were rebred after weaning. Sows were pair-matched by parity and litter average birth weight of the previous three litters. Within pairs, sows were allocated to be fed either standard corn/soyabean meal-based gestation and lactation diets (CON), or the same diets enriched with 0.5% of the mLCPUFA supplement at the expense of corn. Each litter between 9 and 16 total pigs born was classified as LBW or medium/high average birth weight (MHBW) litter and there was a significant correlation (P<0.001) between litter average birth weight of the current and previous litters within sows (r=0.49). Sow serum was harvested at day 113 of gestation for determination of immunoglobulin G (IgG) concentrations. The number of pigs born total and alive were lower (P=0.01) in mLCPUFA than CON sows, whereas the number of stillborn and mummified pigs were similar between treatments. Number of stillborns (trend) and mummies (P<0.01) were higher in LBW than MHBW litters. Tissue weights and brain : tissue weight ratios were similar between treatments, but LBW litters had decreased tissue weights and increased brain : tissue weight ratios compared with MHBW litters. Placental weight was lower (P=0.01) in LBW than MHBW litters, but was not different between treatments. Average and total litter weight at day 1 was similar between treatments. mLCPUFA increased weaning weight (P=0.08) and average daily gain (P<0.05) in MHBW litters, but not in LBW litters. Pre-weaning mortality was similar between treatments, but was higher (P<0.01) in LBW than MHBW litters. IgG concentration in sow serum was similar between treatments and litter birth weight categories. In conclusion, litter birth weight phenotype was repeatable within sows and LBW litters showed the benchmarks of intra-uterine growth retardation (lower placental weight and brain sparing effects). As maternal mLCPUFA supplementation decreased litter size overall, only improved litter growth rate until weaning in MHBW litters, and did not affect pre-weaning mortality, maternal mLCPUFA supplementation was not an effective strategy in our study for mitigating negative effects of a LBW litter phenotype.     

Developmental studies of Hanford miniature swine exposed to 60-Hz electric fields

Evaluations of reproductive and developmental toxicology, including teratology, were included as part of a broad screening study in Hanford Miniature swine (HMS) to detect effects of exposure to electric fields. One group (E) was exposed to a uniform, vertical, 60-Hz, 30-kV/m electric field for 20 h/day, 7 days/week; sham-exposed (SE) swine were housed in a separate, environmentally equivalent building. The first generation (F0) gilts were bred after 4 months of study; some were killed for teratologic assays at 100 days of gestation (dg), and the others produced an F1 generation of offspring. The pooled incidence of terata in these litters (teratologic assays and live births) was similar in the E and SE groups. The F0 females, which produced the F1 generation, were bred again after 18 months of exposure and were killed at 100 dg. Malformation incidence in E litters (75%) was significantly greater than in SE litters (29%). No consistent differences in litter size, fetal mass, or mass of fetal organs were detected. The F1 gilts were bred at 18 months of age; defective offspring were found in significantly more of the E litters (71%) than in SE litters (33%). These F1 females were bred again 10 months later and teratologic assays were performed on their second litters at 100 dg. The percentage of litters with malformed fetuses was essentially identical in the E and SE groups (70% and 73%, respectively). There appears to be an association between chronic exposure to a strong electric field and developmental effects in swine, although the change in incidence of malformations between generations and between the first and second breedings makes it impossible to conclude unequivocally that there is a cause-and-effect relation.     

Production of alpha 1,3-galactosyltransferase gene knockout pigs expressing both human decay-accelerating factor and N-acetylglucosaminyltransferase III

Heterozygous alpha 1,3-galactosyltransferase (GT) gene knockout pigs were produced with transgenic pig fetal cells expressing both human decay-accelerating factor (hDAF) and N-acetylglucosaminyltransferase III (GnT-III). In this study, we assessed the gene targeting efficiency in the transgenic pig fetal cells derived from different fetal tissues such as brain, skin, heart, and liver, or fetal carcass. Targeted cell colonies were selected by hygromycin B. The GT-knockout colonies (KO colonies) were obtained equally from the cells derived from all tissues except liver. Staining with five antibodies against intermediate filaments, all examined KO cell lines stained positive for vimentin with the exception of a colony that stained positive for both vimentin and glial fibrillary acidic protein simultaneously. This is the first study to produce KO cells from the astrocytes. Some of these KO cell lines were used for nuclear transfer (NT) to obtain KO pig fetuses. Fourteen fetuses were obtained from two recipients of the embryo transfer and eight of them had normal ploidy. The cells from the KO pig fetuses were also used for NT to produce cloned KO pigs. Two healthy clone pigs were born. These pigs were determined to have a heterozygous knockout GT gene and the two transgenes. The cells collected from the KO pigs were shown to have similar expression levels of hDAF and GnT-III compared to their original transgenic pigs and less than a half levels of the alphaGal epitopes existed in wild-type pig cells.     

Growth and tissue accretion rates of swine expressing an insulin-like growth factor I transgene

The goal of this research was to determine whether directing expression of an insulin-like growth factor I (IGF-I) transgene specifically to striated muscle would alter the growth characteristics in swine. Transgenic pigs were produced with a fusion gene composed of avian skeletal alpha-actin regulatory sequences and a cDNA encoding human IGF-I. Six founder transgenic pigs were mated to nontransgenic pigs to produce 11 litters of G1 transgenic and sibling control progeny. Birth weight, weaning weight, and proportion of pig survival did not differ between transgenic and control pigs. The ADG of pigs as they grew incrementally from 20 to 60 kg, 60 to 90 kg, and 90 to 120 kg, respectively, did not significantly differ between transgenic and control pigs. Efficiency of feed utilization (gain:feed) was also similar for transgenic and control pigs. Plasma IGF-I and porcine growth hormone (pGH) concentrations were determined at 60, 90, and 120 kg body weight. Plasma IGF-I concentrations were 19% higher in transgenic gilts than control gilts and 11.1% higher in transgenic boars than control boars (P=0.0005). Plasma IGF-I concentrations for boars were also higher than for gilts (P=0.0001). At 60, 90, and 120 kg body weight each pig was scanned by dual energy X-ray absorptiometry (DXA) to derive comparative estimates of carcass fat, lean, bone content of the live animal. Control pigs had more fat and less lean tissue than transgenic pigs at each of the scanning periods and the difference became more pronounced as the pigs grew heavier (P<0.005 at each weight). Transgenic pigs also had a slightly lower percentage of bone than control pigs (P<0.05 at each weight). While daily rates of lean tissue accretion did not differ for transgenic and control pigs, daily rates of fat accretion were lower in transgenic pigs than in control pigs (P<0.05). Based on these results we conclude that expression of IGF-I in the skeletal muscles gradually altered body composition as pigs became older but did not have a major affect on growth performance.     

The effect of exogenous estradiol on litter size in a typical swine herd

Fifty-six female pigs (12 gilts, 26 primiparous and 18 multiparous sows) were bred and assigned to receive either corn oil (vehicle; n=28) or estradiol-17beta (2 mg/day; i.m.; n=28) on Days 12 and 13 (Day 0 = first day of estrus) to determine if exogenous estradiol could improve litter size in swine. Though litter size of the females farrowing was not increased with estradiol treatment (vehicle versus estradiol, 9.7 +/- 0.8 versus 10.1 +/- 1.8 live pigs born, respectively), more females in this group remained pregnant (vehicle versus estradiol, 6 versus 1 recycled, respectively; P<0.05). Litter size, expressed as the number of pigs born per-female-bred, was therefore increased with exogenous estradiol (vehicle versus estradiol, 7.6 +/- 0.8 versus 9.8 +/- 0.7 live pigs born, respectively; P<0.05). Some of these estradiol- treated females produced smaller (P<0.05) litters and this may be of concern in herds experiencing low fertilization/conception rates. Overall 60 more pigs were born alive to females in the estradiol-treated group than to the control group females. This experiment demonstrated that supplemental estradiol treatment on Days 12 and 13 assisted in maintenance of pregnancy and thereby increased litter size on a per-female-bred basis.     

Production of cloned pigs from in vitro systems

Here we describe a procedure for cloning pigs by the use of in vitro culture systems. Four healthy male piglets from two litters were born following nuclear transfer of cultured somatic cells and subsequent embryo transfer. The initiation of five additional pregnancies demonstrates the reproducibility of this procedure. Its important features include extended in vitro culture of fetal cells preceding nuclear transfer, as well as in vitro maturation and activation of oocytes and in vitro embryo culture. The cell culture and nuclear transfer techniques described here should allow the use of genetic modification procedures to produce tissues and organs from cloned pigs with reduced immunogenicity for use in xenotransplantation.     

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.     

Ovulation Statuses of Surrogate Gilts Are Associated with the Efficiency of Excellent Pig Cloning

Somatic cell nuclear transfer (SCNT) is an assisted reproductive technique that can produce multiple copies of excellent livestock. However, low cloning efficiency limits the application of SCNT. In this study, we systematically investigated the major influencing factors related to the overall cloning efficiency in pigs. Here, 13620 cloned embryos derived from excellent pigs were transferred into 79 surrogate gilts, and 119 live cloned piglets were eventually generated. During cloning, group of cloned embryos derived from excellent Landrace or Large white pigs presented no significant differences of cleavage and blastocyst rates, blastocyst cell numbers, surrogate pregnancy and delivery rates, average numbers of piglets born and alive and cloning efficiencies, and group of 101–150, 151–200 or 201–250 cloned embryos transferred per surrogate also displayed a similar developmental efficiency. When estrus stage of surrogate gilts was compared, group of embryo transfer on Day 2 of estrus showed significantly higher pregnancy rate, delivery rate, average number of piglets born, average alive piglet number or cloning efficiency than group on Day 1, Day 3, Day 4 or Day 5, respectively (P<0.05). And, in comparison with the preovulation and postovulation groups, group of surrogate gilts during periovulation displayed a significantly higher overall cloning efficiency (P<0.05). Further investigation of surrogate estrus stage and ovulation status displayed that ovulation status was the real factor underlying estrus stage to determine the overall cloning efficiency. And more, follicle puncture for preovulation, not transfer position shallowed for preovulation or deepened for postovulation, significantly improved the average number of piglets alive and cloning efficiency (P<0.05). In conclusion, our results demonstrated that ovulation status of surrogate gilts was the fundamental factor determining the overall cloning efficiency of excellent pigs, and follicle puncture, not transfer position change, improved cloning efficiency. This work would have important implications in preserving and breeding excellent livestock and improving the overall cloning efficiency.     

A procedure for successful nonsurgical embryo transfer in swine

The current study was undertaken to develop a successful procedure for the nonsurgical transfer of pig embryos. A total of 663 embryos were surgically collected on Day 4 or 5 from 55 donors, of which 542 embryos of acceptable quality were nonsurgically transferred to 46 recipients. Nonsurgical recipient gilts were sedated 15 min prior to transfer with 20 mg im acepromazine maleate. A disposable insemination spirette with an attached 3-way stopcock was manipulated into the cervix of each gilt. Embryos were expelled from a tomcat catheter into the spirette, and 10 to 12 ml of Whitten's medium were used to flush embryos through the spirette into the reproductive tract. Sixteen (34.8%) recipient gilts did not return to estrus before Day 36, and 10 (21.7%) gilts farrowed with an average litter size of 4.3 +/- 0.7. Embryos were collected from an additional 20 donors and were surgically transferred to an additional 19 recipients. Surgical transfers conducted at the same time as the nonsurgical transfers resulted in 12 (63.2%) gilts farrowing and 7.1 +/- 0.6 pigs were born per litter. In conclusion, a procedure has been developed for nonsurgical transfer of swine embryos which simplifies the process of embryo transfer and which may increase the potential for utilization of embryo transfer technologies by swine producers.     

Development rates of male bovine nuclear transfer embryos derived from adult and fetal cells

This study compared the nuclear transfer (NT) embryo development rates of adult and fetal cells within the same genotype. The adult fibroblast cells were obtained from a 21-yr-old Brahman bull. The fetal cells were derived from a Day 40 NT fetus previously cloned using cells from the Brahman bull. Overall, similar numbers of blastocysts developed from both adult (53 of 190; 28%) and fetal (39 of 140; 28%) donor cells. Improved blastocyst development rates were observed when fetal cells were serum-starved (serum-fed 12% vs. serum-starved 43%; P < 0.01) whereas there was no similar benefit when adult cells were serum-starved (both serum-fed and serum-starved 28%). Day 30 pregnancy rates were similar for blastocysts derived from adult (6 of 26; 23%) or fetal (5 of 32; 16%) cells. Day 90 pregnancy rates were 3 of 26 for adult and 0 of 32 for the fetal cell lines. One viable bull calf derived from a 21-yr-old serum-starved adult skin fibroblast was born in August 1999. In summary, somatic NT embryo development rates were similar whether adult or fetal cells, from the same genotype, were used as donor cells. Serum starvation of these adult donor cells did not improve development rates of NT embryos to blastocyst, but when fetal cells were serum-starved, there was a significant increase in development to blastocyst.     

Production of viable pigs from fetal somatic stem cells

Fetal somatic stem cells (FSSCs) are a novel type of somatic stem cells that have recently been discovered in primary fibroblast cultures from pigs and other species. The goal of the present study was to produce viable piglets from FSSCs. NT complexes were prepared from both FSSCs and porcine fetal fibroblasts (pFF) to permit comparison of these two donor cell types. FSSCs from isolated attached colonies were compared with pFF in their ability to form blastocysts upon use in NT. Fusion and cleavage rates were similar between the two groups, while blastocyst rates were significantly higher when using pFF as donor cells. FSSCs of three different size categories derived from dissociation of spheroids yielded similar results. The use of FSSCs of 15-20 microm in size yielded similar cleavage and blastocyst rates as fetal fibroblasts. In the final experiment NT complexes produced from FSSCs were transferred to foster mothers. After transfer to prepubertal gilts, three of seven recipients established pregnancies and delivered seven piglets, of which three piglets were viable and showed normal development. Results for the first time demonstrate that FSSCs are able to produce cloned embryos, and that pregnancies can be established and viable piglets can be produced.     

Production of recombinant human lysozyme in the milk of transgenic pigs

In the swine industry pathogenic infections have a significant negative impact on neonatal survival. Piglets fed with human lysozyme, a natural antibiotic, might be more resistant to gastrointestinal infections. Here we describe the generation of transgenic swine expressing recombinant human lysozyme by somatic cell nuclear transfer. Three cloned female pigs were born, one of which expressed rhLZ at 0.32 ± 0.01 μg/ml in milk, 50-fold higher than that of the pig native lysozyme. Both the transgenic gilts and their progeny appear healthy. Introducing human lysozyme into pigs’ milk has a potential to benefit the piglets by enhancing immune function and defending against pathogenic bacteria, thereby increasing the new born survival rate. This advance could be of great value to commercial swine producers.