Effect of feeding three lysine to energy diets on growth, body composition and age at puberty in replacement gilts.

This study evaluated the effect of diets differing in standard ileal digestible (SID) lysine on lysine intake, growth rate, body composition and age at puberty on maternal line gilts. Crossbred Large White×Landrace gilts (n=641) were fed corn-soybean diets differing in SID lysine concentration (%, g SID lysine:Mcal ME); diets were not isocaloric. Gilts received three grower, finisher diet combinations: low (0.68% lysine grower, 0.52% lysine finisher), medium (0.79% lysine grower, 0.60% lysine finisher) or high (0.90% lysine grower, 0.68% lysine finisher). Grower diets were fed from 100 until 142days of age, and finisher diets were fed until they reached 220days of age. Body weight (BW), backfat thickness (BF), and loin depth (LD) were recorded every 28days. From 160-220days of age, gilts were exposed daily to vasectomized boars and observed for behavioral estrus. Gilts fed the low lysine diet had lower average daily gain and BW (P<0.05), but not fat depth:LD ratio. The percentage of gilts that displayed natural estrus by 220days of age was low but not different among dietary treatments (low 27.7%, medium 31.0% and high 37.7%, respectively; P=0.1201). Gilts fed the high and medium diets reached puberty 10 and 6days earlier, however, than gilts fed the low lysine diet (P<0.05). The rate of puberty attainment may have been less because all gilts contracted porcine epidemic diarrhea (PEDv) just as boar exposure was to begin for the first group of gilts. Results from the present study indicate that growth rate and age at puberty can be altered by ad libitum fed diets that differ in SID lysine concentration.

Relationships between the genes expressed in the mesenteric adipose tissue of beef cattle and feed intake and gain.

Mesenteric fat, a depot within the visceral fat, accumulates in cattle during maturation and finishing and may be a potential source of production inefficiency. The aim of this study was to determine whether the genes expressed in the mesenteric fat of steers were associated with body weight gain and feed intake. Sixteen steers chosen by their rank of distance from the bivariate mean for gain and feed intake were used for this study. Mesenteric fat was obtained and evaluated for differences in gene expression. A total of 1831 genes were identified as differentially expressed among steers with variation in feed intake and gain. Many of these genes were involved with metabolic processes such as proteolysis, transcription and translation. In addition, the Gene Ontology annotations including transport and localization were both over-represented among the differentially expressed genes. Pathway analysis was also performed on the differentially expressed genes. The superoxide radical degradation pathway was identified as over-represented based on the differential expression of the genes GPX7, SOD2 and TYRP1, suggesting a potential role for oxidative stress or inflammatory pathways among low gain-high intake animals. GPX7 and SOD2 were in lower transcript abundance, and TYRP1 was higher in transcript abundance among the low gain-high feed intake animals. The retinoate biosynthesis pathway was also enriched due to the differential expression of the genes AKR1C3, ALDH8A1, RDH8, RDH13 and SDR9C7. These genes were all more highly expressed in the low gain-high intake animals. The glycerol degradation and granzyme A signaling pathways were both associated with gain. Three glycerol kinase genes and the GZMA gene were differentially expressed among high vs. low gain animals. Mesenteric fat is a metabolically active tissue, and in this study, genes involved in proteolysis, transcription, translation, transport immune function, glycerol degradation and oxidative stress were differentially expressed among beef steers with variation in body weight gain and feed intake.

A sandwich ELISA for porcine alpha-1 acid glycoprotein (pAGP, ORM-1) and further demonstration of its use to evaluate growth potential in newborn pigs.

A simple, reproducible sandwich, ELISA was developed to measure porcine alpha-1 acid glycoprotein (pAGP, ORM-1) in pig plasma. Porcine AGP isolated from serum was purchased and a polyclonal antisera was prepared in rabbits using the whole pAGP molecule as immunogen. The antiserum was affinity purified, and a portion of the purified antibody fraction was labeled with horseradish peroxidase. Porcine AGP protein was used as a standard, whereas commercially available buffers and reagents were utilized throughout the assay. The assay was specific for pAGP, had a lower limit of detection of 3.2 ng/mL, and could be used to quantify pAGP in plasma or serum. Using this ELISA, we corroborated our previous findings obtained by RID assay, which demonstrated that the AGP concentration in newborn piglets is negatively associated with preweaning growth rate. The current data were obtained using piglets from a different geographical location and genetic background and showed that elevated AGP at birth was associated with reduced preweaning growth rate (P < 0.001, r = 0.433, n = 19 litters). In addition, litters with a greater average AGP at birth were at a growth disadvantage compared with litters with reduced average AGP plasma concentrations (P < 0.001, r = 0.708, n = 19 litters). Litter average plasma AGP was a better predictor of litter preweaning growth rate than average litter birth weight. The data represent further support for using perinatal AGP concentrations as a tool to identify potential slower growing pigs and as a plasma biomarker for predicting litter growth rate.

Selection for uterine capacity improves lifetime productivity of sows.

Selection for 11 generations for uterine capacity (UC) increased litter size in gilts by 1.6 more fully formed pigs at birth compared to an unselected control line (CO) despite averaging one less ovulation. The objective of the present study was to quantify line by parity interactions and characterize litter performance traits of sows in each line at later parities. Gilts farrowed in contemporary groups of 19 litters and were maintained through four parities if successfully mated in that contemporary group. A total of 243 litters and 2639 piglets were analyzed. Fixed effects of farrowing group, line, parity (1-4), and two-way interactions involving line were fitted. Sire (n=57) of the sow within farrowing group and line was fitted as a random effect. No significant line by parity interactions were observed. Parity effects were detected (P<0.01) for individual piglet birth weight, pre-weaning gain, and weaning weight. Parity effects were also detected (P<0.05) for total number born, average and total litter birth weight, and average and total litter weaning weight. Selection line differences for litter traits were detected (P<0.05) for number stillborn piglets and approached significance (P=0.06) for number of piglets weaned. Retention of sows in the herd was greater (P<0.05) with an average of 2.33 parities for the UC line females compared to 1.87 parities for the CO line. This resulted in favorable cumulative lifetime productivity of the UC line for total number of piglets born, number of piglets born alive, litter birth weight, number of piglets weaned and litter weaning weight.

Litter-of-origin trait effects on gilt development.

The preweaning litter environment of gilts can affect subsequent development. In a recent experiment designed to test the effects of diet on gilt development, litter-of-origin traits including individual birth weights, immunocrits (a measure of colostrum intake), sow parity, number weaned, and individual weaning weights were collected for approximately 1,200 gilts that were progeny of approximately 300 sows. Subsequently, BW, LM area, and backfat were measured at 100 d of age and at 28-d intervals until slaughter (260 d of age). From 160 d of age to slaughter, gilts were observed daily for estrus. At slaughter, the reproductive tract and 1 mammary gland were recovered. The reproductive tract was classified as cyclic or prepubertal; the number of corpora lutea was counted. Uterine horn lengths and ovarian dimensions were measured. Uterus and ovary samples from every 10th gilt were prepared for histological evaluation of uterine gland development and follicle counts, respectively. Mammary gland tissue protein and fat were assayed. Day of the estrous cycle at slaughter was calculated using the first day of the most recent standing estrus (d 0) recorded previous to slaughter. Each gilt development trait was analyzed for association with each litter-of-origin trait, after adjusting for dietary treatment effects. Uterine length, ovarian dimensions, mammary gland protein and fat, and uterine gland development were also adjusted for day of the estrous cycle at slaughter. All litter-of-origin traits were associated ( < 0.05) with growth traits. Top-down (backward elimination) multiple regression analysis indicated that BW and LM accretion in gilts was positively associated with immunocrit ( < 0.01), birth weight ( < 0.01), preweaning growth rate ( < 0.01), and parity ( < 0.01). Backfat accretion was positively associated with preweaning growth rate ( < 0.01), number weaned ( < 0.05), and parity ( < 0.05). Age at puberty was associated with birth weight (positive; < 0.01) and preweaning growth rate (negative; < 0.01). Total uterine length was positively associated with only birth weights ( < 0.05). Mammary gland protein was negatively associated with preweaning growth ( < 0.01). Mammary gland fat was positively associated with birth weight and number of piglets weaned ( > 0.05). These results indicate that colostrum consumption, birth weights, preweaning growth rate, number weaned, and parity are associated with gilt development traits during later life.

Age at puberty, ovulation rate, and uterine length of developing gilts fed two lysine and three metabolizable energy concentrations from 100 to 260 d of age.

The objective of this study was to determine the effect of ad libitum feeding diets differing in standard ileal digestible (SID) lysine and ME concentrations that bracket those fed to developing gilts in U.S. commercial settings. Average SID lysine and ME concentrations in diets currently fed to developing gilts were obtained from a poll of the U.S. commercial swine industry. Crossbred Large White × Landrace gilts (n = 1,221), housed in groups, were randomly allotted to 6 corn-soybean diets in a 2 × 3 factorial arrangement formulated to provided 2 SID lysine and 3 ME concentrations. Gilts received grower diets formulated to provide 1.02% (control = survey average) or 0.86% (control minus 15%) SID lysine and 2.94, 3.25, or 3.57 (survey average ME ± 10%) Mcal of ME/kg from 100 d of age until approximately 90 kg BW. Then, gilts were fed finisher diet containing 0.85% (control = survey average) or 0.73% (control minus 15%) SID lysine and 2.94, 3.26, or 3.59 (control ± 10%) Mcal of ME/kg until 260 d of age. Gilts were weighed, and backfat thickness and loin muscle area were recorded at the beginning of the trial and then every 28 d. Starting at 160 d of age, gilts were exposed daily to vasectomized boars and observed for behavioral estrus. At approximately 260 d of age, gilts were slaughtered and their reproductive tract was collected. Each reproductive tract was examined to determine whether the gilt was cyclic, the stage of estrus cycle, ovulation rate, and uterine length. Data were evaluated for normality and analyzed using mixed model methods. Average age at puberty was 193 d of age with a range from 160 to 265 d. When all gilts on trial at 160 d of age were included in the analysis, 91.0% reached puberty as determine by observation of standing estrus. Differences between dietary treatments on age at puberty or measurements of the reproductive tract were not detected. Growth rates to 160 d were not limiting for attainment of puberty in response to daily boar stimulation from 160 d.

Relationships between day one piglet serum immunoglobulin immunocrit and subsequent growth, puberty attainment, litter size, and lactation performance.

Colostrum affects gut and uterine gland development in the neonatal piglet, suggesting that subsequent growth and reproductive performance may be affected. Measuring immunoglobulin in piglet serum using the immunoglobulin immunocrit on Day 1 of age provides a simple, inexpensive indication of the amount of colostrum acquired by the piglet in the first day of life. Relationships between serum immunoglobulin immunocrit measures and subsequent growth rates, age at puberty, incidence of puberty failure, litter size, and lactation performance were examined in pigs born and subsequently farrowing between 2009 and 2013. Immunoglobulin immunocrit measures were collected on 16,762 piglets on Day 1 of age. Of these piglets, BW measurements were available from 15,324 (7,684 males and 7,640 females) piglets at a range of ages from weaning to 200 d of age, allowing an assessment of growth rates. Age at puberty was recorded from a subset of 2,857 of the females after observing them for estrous behavior from approximately 170 to 250 d of age. To examine relationships between d 1 immunocrit and puberty failure, gilts with immunocrit measures that failed to reach puberty (n = 119) were matched with littermate gilts with immunocrit measures that achieved puberty (n = 167). Similarly, number born alive was collected on a subset (n = 799) of females from first to fourth parities for which d 1 immunocrits were measured on them as neonates. Finally, d 1 immunocrit effect on adult lactational competence was assessed by measuring litter average (offspring of 440 females) and litter average piglet preweaning growth rate (offspring of 774 females) in females where d 1 immunocrits were available from them as neonates. Results indicated that low d 1 immunocrits were subsequently associated with reduced growth (P < 0.01), increased age at puberty (P < 0.01), reduced number born alive (P < 0.05), reduced litter average immunocrit (P < 0.05), and reduced litter average preweaning growth rate during lactation (P < 0.05). This suggests that management efforts to improve the amount of colostrum ingested by neonatal piglets would result in beneficial changes in production efficiency, particularly for gilts destined for the breeding herd. It also suggests that the immunoglobulin immunocrit can be useful in monitoring colostrum ingestion to maximize the beneficial effects of colostrum on subsequent performance.

Optimal dietary energy and amino acids for gilt development: Growth, body composition, feed intake, and carcass composition traits.

The objective of this study was to determine if body composition of developing gilts could be altered at the onset of estrus by ad libitum feeding diets differing in standard ileal digestible (SID) lysine and ME using levels that are within those used in practice by pig producers in the United States. Crossbred Large White × Landrace gilts ( = 1,221), housed in groups, were randomly allotted to 6 corn-soybean diets in a 2 × 3 factorial arrangement formulated to provide 2 SID lysine and 3 ME levels. Gilts received grower diets formulated to provide 0.86 (low) or 1.02% (high) SID lysine and 2.94 (low), 3.25 (medium), or 3.57 (high) Mcal of ME/kg from 100 d of age until approximately 90 kg BW. Then, gilts were fed finisher diets containing 0.73 (low) or 0.85% (high) SID lysine and 2.94 (low), 3.26 (medium) or 3.59 (high) Mcal of ME/kg until 260 d of age. The medium SID lysine and medium-ME diets were based on an informal survey from the U.S. commercial swine industry to obtain average levels that are currently being formulated for developing gilts. Gilts were weighed and backfat thickness and loin area were recorded at the beginning of the trial and then every 28 d. Feed intake (FI) was recorded as feed disappearance within the pen at 2-wk intervals. Lysine (g) and ME (Mcal) consumed were calculated based on diet formulations. At approximately 260 d of age, gilts were slaughtered and warm carcass weight and fat thickness were recorded. There were no differences between lysine or ME levels for growth and body composition, except for backfat, which was slightly greater for gilts fed a high-ME diet. Gilts fed high-ME diets had a lower FI but a greater ME intake compared with gilts fed low ME ( < 0.05). Additionally, gilts fed the high-ME diet had lower FI and lysine intake per kilogram of BW gain when compared with gilts fed low- or medium-ME diets ( < 0.05). However, there was no difference in the megacalories consumed per kilogram of BW gain among treatments ( > 0.05). Carcasses from gilts fed the high-ME diet were 3.3 and 2.5 kg heavier than those from gilts fed the low- or medium-ME diets ( < 0.05). Despite significant differences in the lysine:ME ratio in the diets, no changes in growth or body composition occurred, likely due to compensatory changes in FI in response to dietary ME content. Caloric efficiency (Mcal to deposit 1 kg of BW) was similar among treatments.

Contributions of the maternal uterine environment and piglet genotype on weaning survivability potential: II. Piglet growth, lactation performance, milk composition, and piglet blood profiles during lactation following reciprocal embryo transfers between Meishan and White crossbred gilts.

Crossbreeding studies between Meishan (MS) and Large White (LW) pigs have illustrated that increased piglet growth before weaning is attributed to the maternal genotype of LW dams. The objective of this study was to determine the contributions of the maternal uterine environment (MUE), piglet genotype (PigG), piglet age (PA), and their interactions on piglet growth, lactation performance, milk composition, and piglet blood profiles during lactation following reciprocal embryo transfers between MS and White crossbred (WC) gilts. Twenty-five successful pregnancies were generated by embryo transfer in 2 farrowing years representing all MUE × PigG combinations: MS × MS (n = 4 litters), MS × WC (n = 7 litters), WC × MS (n = 7 litters), and WC × WC (n = 7 litters). At d 1 and 10 and at weaning, piglets (n = 147, n = 96, and n = 94, respectively) were weighed and blood samples were collected and measured for hematocrit, hemoglobin, glucose, nitrogen, NEFA, albumin, lactate, and cortisol. In addition, sows were manually milked from a medial mammary gland to determine milk composition. All data were analyzed by ANOVA using MIXED model procedures with the fixed effects of MUE, PigG, PA, and their interactions. Piglet weight was greater (P < 0.001) in piglets from WC dams compared to MS dams at d 10 and weaning but not at d 1. In addition, ADG were greater (P < 0.05) from piglets from WC dams compared to MS dams throughout lactation. However, milk composition was greater (P < 0.05) for GE and fat content from MS dams compared to WC dams, illustrating differences in milk quality between the breeds. There were significant MUE × PigG × PA interactions for hematocrit and hemoglobin levels in which greater (P < 0.001) levels were observed in MS piglets, irrespective of MUE, at d 1 of lactation and in MS piglets from MS dams at d 10 of lactation. Blood glucose was greater (P = 0.01) at d 1 in piglets from WC dams regardless of PigG but, at weaning, glucose was greater (P = 0.01) in WC piglets regardless of MUE. Serum NEFA levels were greater (P = 0.02) in piglets from MS dams throughout the lactation period. This study demonstrated that WC dams were superior to MS dams for piglet growth during lactation, in agreement with previous crossbreeding studies. However, blood components measured displayed complex interactions between the piglet and maternal breed, which signify possible mechanisms for improved preweaning survivability but slower lactational growth of MS piglets.

Genomewide association analysis for average birth interval and stillbirth in swine.

Reproductive efficiency has a great impact on the economic success of pork production. Stillborn pigs and average birth interval contribute to the number of pigs born alive in a litter. To better understand the underlying genetics of these traits, a genomewide association study was undertaken. Samples of DNA were collected and tested using the Illumina Porcine SNP60 BeadChip from 798 females farrowing over a 4-yr period (all first parity). Birth intervals and piglet birth status (stillborn or alive) were determined by videotaping each farrowing event. A total of 41,148 SNP were tested using the Bayes C option of GenSel (version 4.61) and 1-Mb windows. These 1-Mb windows explained proportions of 0.017, 0.002, 0.032, 0.029, and 0.030 of the total variation, respectively, for litter average birth interval after deletion of the last piglet born, last birth interval in the litter, number of stillborn piglets ignoring the last piglet born, number of stillborns in the last birth position, and percent stillborn ignoring the last piglet. Significant 1-Mb nonoverlapping SNP windows were identified by using a conservative approach requiring 1-Mb windows to have a genetic variance ≥1.0% of genomic variance and these were considered to be QTL. Quantitative trait loci were located for number of stillborn piglets ignoring the last piglet born (1 QTL), number of stillborns in the last birth position (1 QTL), and percent stillborn ignoring the last piglet (3 QTL). In addition, 2, 13, 3, and 6 suggestive 1-Mb nonoverlapping SNP windows were identified for litter average birth interval after deletion of the last piglet born, number of stillborn piglets ignoring the last piglet born, number of stillborns in the last birth position, and percent stillborn ignoring the last piglet, respectively. Possible candidate genes affecting both birth interval and stillbirth included () and (). Possible genes affecting only birth interval included (), and (), and those affecting only stillbirth included (), LOC100518697 (a nostrin-like gene), and (). The QTL and the suggestive 1-Mb nonoverlapping SNP windows may lead to genetic markers for marker assisted selection, marker assisted management, or genomic selection applications in commercial pig populations.

Genomewide association and identification of candidate genes for ovulation rate in swine.

Reproductive efficiency has a great impact on the economic success of pork production. Ovulation rate is an early component of reproduction efficiency and contributes to the number of pigs born in a litter. To better understand the underlying genetics of ovulation rate, a genomewide association study was undertaken. Samples of DNA were collected and tested using the Illumina Porcine SNP60 BeadChip from 1,180 females with ovulation measurements ranging from never farrowed to measurements taken after parity 2. A total of 41,848 SNP were tested using the Bayes C option of GenSel. After the Bayes C analysis, SNP were assigned to sliding windows of 5 consecutive SNP by chromosome-position order beginning with the first 5 SNP on SSC1 and ending with the last 5 SNP on SSCX. The 5-SNP windows were analyzed using the Predict option of GenSel. From the Predict analysis, putative QTL were selected having no overlap with other 5-SNP window groups, no overlap across chromosomes, and the highest genetic variation. These putative QTL were submitted to statistical testing using the bootstrap option of GenSel. Of the putative QTL tested, 80 were found to be statistically significant (P < 0.01). Ten QTL were found on SSC1, 12 on SSC2, 4 on SSC3, 8 on SSC4, 3 on SSC5, 3 on SSC6, 3 on SSC7, 4 on SSC8, 2 on SSC9, 4 on SSC10, 1 on SSC12, 4 on SSC13, 2 on SSC14, 4 on SSC15, 4 on SSC16, 6 on SSC17, 4 on SSC18, and 1 on SSCX. Sixteen QTL were found to be statistically significant at the P < 0.001 level. Six additional QTL were significant at the P = 0.001 level. These 22 QTL accounted for 71.10% of the total genetic variance. The most compelling candidate genes in these regions include Estrogen receptor 1, growth differentiation factor 9, and inhibin ßA. These QTL, when combined with information on genes found in the same regions, should provide useful information that could be used for marker assisted selection, marker assisted management, or genomic selection applications in commercial pig populations.

Effect of essential fatty acid and zinc supplementation during pregnancy on birth intervals, neonatal piglet brain myelination, stillbirth, and preweaning mortality.

Omega fatty acids and zinc contribute to physiological pathways that could affect the farrowing process, stillbirth, preweaning mortality, and postweaning return to estrus. To determine effects of omega fatty acids and zinc on these reproductive traits, gilts were mated and fed either a control diet, a diet supplemented with 1.09% Gromega, a diet supplemented with 0.07% zinc sulfate, or a diet supplemented with both Gromega and zinc sulfate from d 80 of gestation until farrowing. Farrowings were video recorded to obtain birth intervals for each piglet, and the number of live and stillborn piglets was recorded. On d 1 after farrowing, piglets were weighed, and the smallest piglet in each litter was sacrificed. A blood sample was collected to measure the immunoglobulin immunocrit ratio, and brain, cerebellum, brain stem, full and empty stomach (to calculate stomach content weight), and heart weights were recorded. Because myelination of specific brain regions may affect preweaning mortality, brain stem, cerebellum, and spinal cord tissues were measured for content of myelin basic proteins and myelin lipids. For remaining piglets, survival to weaning and weaning weights were recorded. Results indicated a weak positive correlation (r = 0.23, P < 0.05) between immunocrit values and brain stem high molecular weight myelin basic protein. There was also a Gromega × zinc supplementation interaction (P < 0.05) on brain stem high molecular weight myelin basic protein in which the combined treatment was greater than the control or each supplement alone. Zinc treatment decreased stillbirth rate during prolonged farrowing and subsequent preweaning survival of low birth weight piglets. Gromega increased overall stillbirth rate and increased the stillbirth rate during prolonged farrowing. There were no relationships between myelin measurements and preweaning survival. In conclusion, combined Gromega and zinc supplementation appeared to improve myelination, but zinc alone improved stillbirth and preweaning survival.

Identifying genetic loci controlling neonatal passive transfer of immunity using a hybrid genotyping strategy.

Colostrum intake is critical to a piglet's survival and can be measured by precipitating out the γ-immunoglobulins from serum with ammonium sulfate (immunocrit). Genetic analysis of immunocrits on 5312 piglets indicated that the heritabilities (se) for direct and maternal effects were 0.13 (0.06) and 0.53 (0.08) respectively. To identify QTL for direct genetic effects, piglets with the highest and lowest immunocrits from 470 litters were selected. Six sets of DNA pools were created based on sire of the litter. These 12 DNA pools were applied to Illumina Porcine SNP60 BeadChips. Normalized X and Y values were analyzed. Three different SNP selection methods were used: deviation of the mean from high vs. low pools, the deviation adjusted for variance based on binomial theory and ANOVA. The 25 highest ranking SNPs were selected from each evaluation for further study along with 12 regions selected based on a five-SNP window approach. Selected SNPs were individually genotyped in the 988 piglets included in pools as well as in 524 piglets that had intermediate immunocrits. Association analyses were conducted fitting an animal model using the estimated genetic parameters. Nineteen SNPs were nominally associated (P < 0.01) with immunocrit values, of which nine remained significant (P < 0.05) after Bonferroni correction, located in 16 genomic regions on 13 chromosomes. In conclusion, the pooling strategy reduced the cost to scan the genome by more than 80% and identified genomic regions associated with a piglet's ability to acquire γ-immunoglobulin from colostrum. Each method to rank SNPs from the pooled analyses contributed unique validated markers, suggesting that multiple analyses will reveal more QTL than a single analysis.

Genome-wide association with delayed puberty in swine.

An improvement in the proportion of gilts entering the herd that farrow a litter would increase overall herd performance and profitability. A significant proportion (10-30%) of gilts that enter the herd never farrow a litter; reproductive reasons account for approximately a third of gilt removals, with anestrous and failure to conceive the most common reasons for culling. Tools to select gilts for reproductive longevity through genomics or alternative phenotypes would be of great benefit to the producer. Ninety-one gilts that failed to display behavioral estrus by 240 days (cases) and 127 pubertal littermates (controls) were genotyped with the Illumina Porcine SNP60 Beadchip. One hundred and seventy-four SNPs with the most significant associations were genotyped in an additional 86 cases and 103 controls. Twelve of these associations were significant in the final analysis. The most significant (P < 1.5 × 10(-14) ) region associated with failure to attain puberty was on chromosome 4 surrounding the NHLH2 gene. Delayed pubertal development and age at first estrus have been associated with NHLH2 in mice. Because attainment of puberty is a complex trait, identifying genes that affect pubertal age would greatly contribute to our knowledge of reproductive development as well as overall fertility.

Triennial Reproduction Symposium: limitations in uterine and conceptus physiology that lead to fetal losses.

Conceptus losses in livestock occur throughout gestation. The uterus and the embryo-placenta-fetus play interconnected roles in these losses, the details of which depend on the period of gestation and the species. Studies in sheep and pigs have indicated that the uterine glands are essential for full fertility, based on experiments where gland development was reduced through the use of exogenous hormones. In sheep and cattle, normally the uterus is well able to support more than a single fetus although these species differ in the consequences of multiple births. When 2 conceptuses are present, the placentas of cattle often anastomose, putting 1 fetus at risk if the other is lost. One likely reason this does not occur in sheep is because sheep embryos undergo intrauterine migration, similar to pigs. In pigs, the relatively equidistant separation of conceptuses is likely to be essential for optimizing conceptus survival as is the simultaneous and uniform elongation of blastocysts that occurs during the time of maternal recognition of pregnancy. Other studies in pigs have indicated that the size of the uterus influences litter size and therefore fetal losses. In response to crowded intrauterine conditions in the pig, increased conceptus losses begin to occur between d 30 and 40 of pregnancy, and further losses occur sporadically during later gestation. There is evidence that improved fetal erythropoiesis can reduce these losses. Other studies indicated that profound changes in placental development occurred under crowded intrauterine conditions that may contribute to losses during late gestation. Reductions in placental stroma formation may compromise the ability of the pig placenta to adapt to reduced uterine space. Consistent with this, both hyaluronan and hyaluronidase activity are decreased in the placentas of small compared with large fetuses. These results indicate that improvements in placental stroma formation could improve placental ability to compensate for reduced intrauterine space, resulting in increased placental function and reduced fetal losses during late gestation.

Effect of creatine supplementation during the last week of gestation on birth intervals, stillbirth, and preweaning mortality in pigs.

We hypothesized that creatine supplementation would reduce birth intervals, stillbirth rate, and preweaning survival in pigs because of its reported improvement of athletic performance in humans. In Exp. 1, gilts (n = 42) and first parity sows (n = 75) were mated at estrus. Beginning on d 110 of gestation, dams received either no treatment or 20 g creatine daily until farrowing. At farrowing in November 2008, pigs were monitored by video camera to determine individual piglet birth intervals. On d 1, piglets were weighed, euthanized, and the cerebellum, brain stem, and spinal cord were collected from the largest and smallest piglets in each litter to measure myelin basic proteins, myelin cholesterol, glucocerebrosides, phosphatidylethanolamine, phosphatidylcholine, and sphingomyelin. Preweaning mortality of the remaining piglets was recorded, including whether a piglet had been overlayed by the dam. A second experiment was performed using gilts (n = 90), farrowing in July 2010, to test differential effects of creatine supplementation during hot, humid weather when dams typically have more difficulty farrowing. Once again, gilts were provided either no supplementation or 20 g creatine daily from d 110 to the day of farrowing. Gilts were video recorded during farrowing, piglets were weighed on d 1, and preweaning mortality (including overlays) was recorded. In Exp. 1, creatine supplementation had no effect on birth intervals or stillbirth rate. Creatine supplementation improved the amount of myelin lipids in brain regions of piglets, particularly the brain stem. Creatine supplementation also reduced overlays of low birth weight piglets from gilts but not second parity sows. Data from Exp. 2 were combined with gilt data from Exp. 1 to examine the effect of creatine, season, and their interaction. There were no effects of treatment or season on birth intervals, stillbirth rates, or overall preweaning mortality. Creatine treatment reduced the incidence of overlays in low birth weight piglets in the combined data set. These results suggest that creatine supplementation improved myelination and may reduce the incidence of low birth weight piglets being crushed by the dam.

A simple novel measure of passive transfer of maternal immunoglobulin is predictive of preweaning mortality in piglets.

Two factors that contribute to preweaning mortality in piglets are the initiation of lactation by sows and their ability to nurse their piglets. The objective of this study was to determine if the quantification of the transfer of immunoglobulins (Igs) from sow to piglet could act as a measure of these sow factors in terms of their influence on preweaning mortality. To measure passive transfer, a simple, rapid Ig immunocrit method was developed. For validation, the smallest piglets from 204 gilts were sacrificed on day 1 after birth and blood was collected. Piglet serum Ig concentrations were measured three ways: (1) by protein A sepharose precipitation, SDS-PAGE, and densitometry of the IgG heavy chain; (2) by precipitation of Ig with (NH(4))(2)SO(4) followed by spectrophotometric measurement; and (3) by precipitation of Ig with (NH(4))(2)SO(4) and measurement of the ratio of precipitate to sample volume using a hematocrit microcapillary (the Ig immunocrit method). Results from the (NH(4))(2)SO(4) methods correlated (r=0.86) with those obtained using SDS-PAGE. Day 1 weights and immunocrit ratios and preweaning survival data were then collected from every piglet from first (n=90), and second (n=145) parity sows. Bodyweight and immunocrit ratios accounted for 4.8% and 16.7% (P<0.01) of the variation in preweaning mortality, respectively. Litter average immunocrit ratios were not correlated with preweaning mortality. In conclusion, the Ig immunocrit method is a simple, rapid measure of passive transfer from sow to piglet, and is useful in assessing the initiation of colostrum and the nursing ability of sows, and the effect of these events on preweaning piglet mortality.

Lactation Biology Symposium: lactocrine signaling and developmental programming.

Lactocrine signaling is defined as transmission of bioactive factors from mother to offspring as a consequence of nursing. Lactocrine transmission of signaling molecules may be an evolutionarily conserved process through which bioactive factors necessary for support of neonatal development are delivered postnatally. Dependence on maternal resources for development in eutherian mammals extends into neonatal life for at least that period of time when nutrition is obtained solely from first milk (i.e., colostrum). Data for the pig (Sus scrofa domesticus) provide evidence of lactocrine mediated effects on development of the female reproductive tract and other somatic tissues. Porcine uterine gland development, an estrogen receptor-alpha (ESR1)-dependent process, begins within 2 d of birth [postnatal day (PND) 0]. A lactocrine-driven, ESR1-mediated process was proposed as a regulatory mechanism governing onset of uterine gland development and endometrial maturation in the neonatal pig. Gilts maintained in a lactocrine-null state for 2 d from birth by milk-replacer feeding displayed altered patterns of endometrial gene expression and retarded uterine gland development by PND 14. In lactocrine-null gilts, inhibition of endometrial and cervical ESR1 and vascular endothelial growth factor (VEGFA) expression observed on PND 2 persisted to PND 14, even after gilts were returned to nursing on PND 2. Collectively, data support a role for lactocrine signaling in regulation of critical neonatal developmental events. Maternal lactocrine programming of postnatal development may help to insure healthy developmental outcomes. A systems biology approach will be required to define and understand mechanistic dynamics of lactocrine signaling events that may ultimately connect genotype to phenotype and establish the parameters of reproductive potential.

Assessment LOPU-IVF in Japanese sika deer (Cervus nippon nippon) and application to Vietnamese sika deer (Cervus nippon pseudaxis) a related subspecies threatened with extinction.

In mammals, recovery of oocytes by laparoscopic ovum pick-up (LOPU) coupled with in vitro production (IVP) of embryos represents a promising strategy for both amplification and genetic management of sparse animals from captive endangered wild species. As integrated technique developed mainly for domestic livestock, LOPU-IVP requires several studies to set up protocols for follicular stimulation or optimization of IVP before envisaging successful transposition to wild species. In deer, many endangered subspecies would be potentially concerned by applying such an approach using common subspecies for protocols optimization. The aim of the present study was to assess efficiency of follicle stimulation using ovine FSH (oFSH) for recovery of oocytes by LOPU in common sika deer (Cervus nippon nippon) before transposition of an optimized methodology for IVP of embryos from endangered Vietnamese sika deer hinds (Cervus nippon pseudaxis). In common sika deer, two doses of oFSH (0.25 and 0.5 U) and two frequencies of administration (12 and 24 h) were compared by monitoring of subsequent ovarian response, quality of oocytes recovered by LOPU, and in vitro developmental competence. In a first experiment, the dose of oFSH administered did not significantly affect the total number of follicles aspirated per hind per session (8.6 ± 1.0 vs. 8.2 ± 1.6 with 0.5 vs. 0.25 U oFSH, respectively; not significant). In a second experiment, frequency of 0.25 U oFSH administration did not affect ovarian response. Efficiency of IVP determined on blastocysts rates after in vitro maturation, fertilization, and development in oviduct epithelial cells coculture was increased when FSH was administered at 12-h intervals. Immune response after several follicular stimulations was detected against exogenous oFSH in plasma from the majority of sika deer hinds but was not associated with decreased ovarian response. When 0.25 U oFSH was administered at 12-h intervals to Vietnamese sika deer (N = 4), good quality cumulus oocyte complexes with complete and compact cumulus investments were recovered allowing a high cleavage rate after in vitro maturation and fertilization. Development to the blastocyst stage occurred in a high proportion (30% of oocytes) after coculture with ovine epithelial cells allowing cryobanking of transferable embryos from Vietnamese sika deer. These results confirm that LOPU-IVF after ovarian stimulation with oFSH may be a successful tool for cryobanking transferable embryos from endangered sika deer subspecies.

Comparison of myelination between large and small pig fetuses during late gestation.

We compared myelination of the cerebellum, brain stem, and spinal cord in the largest and smallest pig fetuses within a litter during late gestation. Gilts were killed on Days 92, 100, and 110 of gestation and these neural tissues were obtained from the largest and smallest fetuses in each litter. Myelin basic protein (MBP) mRNA was quantified in each tissue using real time reverse transcriptase polymerase chain reaction (rtPCR). Myelin was recovered from each tissue and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and thin layer chromatography (TLC) was used to measure MBP and lipids, respectively. MBP mRNA increased with advancing gestation in all three tissues examined (P≤0.05) and was less in brain stem of small piglets compared to large piglets (P<0.01). Two coomassie stained protein bands (HMBP and LMBP) were observed by SDS-PAGE. Six prominent lipid bands were obtained by TLC (cholesterol, hydroxy(h)-cerebroside, nonhydroxy(nh)-cerebroside, phosphatidylethanolamine, phosphatidylcholine, and sphingomyelin). Significant day by fetal size interactions for cerebellar MBP and lipids indicated that cerebellar myelination in the smallest fetuses was less compared to the largest fetuses on Days 100 and 110 of gestation. Myelin MBP and lipid obtained from brain stem increased with advancing gestation and LMBP and lipids were less in small piglets compared to large piglets. In contrast, myelination in spinal cord increased with day of gestation but was not different between smallest and largest fetuses. These results confirm that myelination of the cerebellum and brain stem, but not spinal cord, is reduced in small fetuses during late gestation.