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The goal of this experiment was to measure the physiological response of individual laying hens exposed to heat stress (HS). Performance, egg quality, body temperature (BT), and blood chemistry of laying hens were individually recorded before and after various intervals of daily cyclic HS. In total, 407 18-week-old W-36 parent-line laying hens (Hy-Line International, Dallas Center, IA) were housed individually in battery cages. After an acclimation period, baseline data were collected from 22 to 24-wk before the hens were subjected to a daily cyclic HS consisting of 7 h at 35°C returning to 30°C for the remaining 17 h/D from 24 to 28-wk of age. Eggs were collected and individually weighed daily. Feed intake (FI), egg production (EP), egg weights, egg mass, BW, and feed efficiency (FE) (g egg/kg FI) were calculated over 2-wk time periods. Eggs were collected for quality assessment the day before HS began, the 2nd day of HS, and on a weekly basis throughout the 4-wk HS. Blood was collected and BT measured the day before heat HS was initiated, on the first day of HS, and again at 2 and 4-wk of HS. Blood PCO2and iCa decreased, and blood pH increased within 4 to 6 h of HS (P≤ 0.01). Shell weights decreased with acute HS, possibly due to the reduction in blood iCa (P≤ 0.01). After 4-wk of HS the blood pH returned to pre-HS levels but iCa remained decreased (P≤ 0.01). Shell weights remained low and Haugh units decreased after 2 and 4-wk of HS (P≤ 0.01). Feed efficiency was increased and FI, EP, and BW decreased by 2-wk of HS and remained low through 4-wk (P≤ 0.01). The cyclic HS had a significant effect on the performance, egg quality, and blood chemistry over the 4-wk HS.

Feet and legs of replacement females were objectively evaluated at selection, i.e., approximately 150 d of age (n= 319) and post first parity, i.e., any time after weaning of first litter and before second parturition (n= 277) to 1) compare feet and leg joint angle ranges between selection and post first parity; 2) identify feet and leg joint angle differences between selection and first 3 wk of second gestation; 3) identify feet and leg joint angle differences between farms and gestation days during second gestation; and 4) obtain genetic variance components for conformation angles for the two time points measured. Angles for carpal joint (knee), metacarpophalangeal joint (front pastern), metatarsophalangeal joint (rear pastern), tarsal joint (hock), and rear stance were measured using image analysis software. Between selection and post first parity, significant differences were observed for all joints measured (P< 0.05). Knee, front and rear pastern angles were less (more flexion), and hock angles were greater (less flexion) as age progressed (P< 0.05), while the rear stance pattern was less (feet further under center) at selection than post first parity (only including measures during first 3 wk of second gestation). Only using post first parity leg conformation information, farm was a significant source of variation for front and rear pasterns and rear stance angle measurements (P <0.05). Knee angle was less (more flexion; P <0.05) as gestation age progressed. Heritability estimates were low to moderate (0.04–0.35) for all traits measured across time points. Genetic correlations between the same joints at different time points were high (>0.8) between the front leg joints and low (<0.2) between the rear leg joints. High genetic correlations between time points indicate that the trait can be considered the same at either time point, and low genetic correlations indicate that the trait at different time points should be considered as two separate traits. Minimal change in the front leg suggests conformation traits that remain between selection and post first parity, while larger changes in rear leg indicate that rear leg conformation traits should be evaluated at multiple time periods.

Heat stress is one of the limiting factors negatively affecting pig production, health, and fertility. Characterizing genomic regions responsible for variation in HS tolerance would be useful in identifying important genetic factor(s) regulating physiological responses to HS. In the present study, we performed genome-wide association analyses for respiration rate (RR), rectal temperature (TR), and skin temperature (TS) during HS in 214 crossbred gilts genotyped for 68,549 single nucleotide polymorphisms (SNP) using the Porcine SNP 70K BeadChip. Considering the top 0.1% smoothed phenotypic variances explained by SNP windows, we detected 26, 26, 21, and 14 genes that reside within SNPs explaining the largest proportion of variance (top 25 SNP windows) and associated with change in RR (ΔRR) from thermoneutral (TN) conditions to HS environment, as well as the change in prepubertal TR(ΔTR), change in postpubertal ΔTR, and change in TS(ΔTS), respectively. The region between 28.85 Mb and 29.10 Mb on chromosome 16 explained about 0.05% of the observed variation for ΔRR. The growth hormone receptor (GHR) gene resides in this region and is associated with the HS response. The other important candidate genes associated with ΔRR (PAIP1, NNT, and TEAD4), ΔTR(LIMS2, TTR, and TEAD4), and ΔTS(ERBB4, FKBP1B, NFATC2, and ATP9A) have reported roles in the cellular stress response. The SNP explaining the largest proportion of variance and located within and in the vicinity of genes were related to apoptosis or cellular stress and are potential candidates that underlie the physiological response to HS in pigs.

Intramuscular fat (IMF) content is an important trait affecting the quality of pork. Two Duroc populations, one under positive selection for IMF and the other selected for decreased backfat but under stabilizing selection for IMF, were used to identify signatures of selection associated with IMF using 60,000 single-nucleotide polymorphism data. The effects of selection were analyzed between 2 lines or groups representing selected and control animals within each population using a discriminant analysis of principal components and Wright's fixation index (FST). Moreover, extended haplotype homozygosity-based approaches were used to examine the changes in haplotype frequency due to recent selection. Each statistical method identified 10–20 selection signatures. A few haplotype-based signatures of selection agreed with results from a genome-wide association study (GWAS), while FSTmeasures showed a better agreement with GWAS results. Agreement of marker-trait associations and signatures of selection was limited, and further examination will be necessary to understand the effect of selection on IMF and why some regions identified by GWAS did not appear to respond to the selection practiced. The genes in 21 consensus selection signatures were examined. Several genes with an effect on overall fatness were identified, but further research is needed to assess whether or not some of them could have a specific effect on IMF.

Marbling and intramuscular fat (IMF) content are commonly used to describe or measure intramuscular fat deposition in meat, which contributes to taste, texture and flavor. Four types of genetic markers, i.e., microsatellite, random amplification of polymorphic DNA (RAPD), amplified fragment length polymorphisms (AFLP) and single nucleotide polymorphisms (SNPs) have been used in genome scans or association studies to detect quantitative trait loci (QTL) for these traits in cattle and swine. For the most part microsatellite markers help define QTL regions but have been used in limited ways to patent gene tests because of the uncertainties associated with the microsatellite marker scans. However, SNPs in candidate genes selected based on physiological, positional or comparative information often lead to patent applications once strong associations have been determined. To date, at least 22 patents have been awarded or under review for genes/markers affecting marbling or IMF in cattle and swine. Unfortunately, similar muscle lipid accumulation in humans has significant negative impacts on health, causing obesity/type 2 diabetes and their associated conditions. Many studies have also been performed on human subjects or on the mouse as a model organism to understand the genetic complexity of these conditions. A collection of over 2,000 reports on genes/markers affecting fat phenotypes in humans, mice, cattle and swine have led to construction of a mammalian concordant QTL map for lipogenesis. The concordant QTL map provides power for fine mapping and narrowing each of these QTL regions to a few genes.

The relationships between quantitative and reproductive fitness traits in animals are of general biological importance for the development of population genetic models and our understanding of evolution, and of great direct economical importance in the breeding of farm animals. Two well investigated quantitative traits—body weight (BW) and litter size (LS)—were chosen as the focus of our review. The genetic relationships between them are reviewed in fishes and several mammalian species. We have focused especially on mice where data are most abundant. In mice, many individual genes influencing these traits have been identified, and numerous quantitative trait loci (QTL) located. The extensive data on both unselected and selected mouse populations, with some characterized for more than 100 generations, allow a thorough investigation of the dynamics of this relationship during the process of selection. Although there is a substantial positive genetic correlation between both traits in unselected populations, caused mainly by the high correlation between BW and ovulation rate, that correlation apparently declines during selection and therefore does not restrict a relatively independent development of both traits. The importance of these findings for overall reproductive fitness and its change during selection is discussed.

Data from an F2 cross between breeds of livestock are typically analysed by least squares line-cross or half-sib models to detect quantitative trait loci (QTL) that differ between or segregate within breeds. These models can also be combined to increase power to detect QTL, while maintaining the computational efficiency of least squares. Tests between models allow QTL to be characterized into those that are fixed (LC QTL), or segregating at similar (HS QTL) or different (CB QTL) frequencies in parental breeds. To evaluate power of the combined model, data wih various differences in QTL allele frequencies (FD) between parental breeds were simulated. Use of all models increased power to detect QTL. The line-cross model was the most powerful model to detect QTL for FD>0·6. The combined and half-sib models had similar power for FD<0·4. The proportion of detected QTL declared as LC QTL decreased with FD. The opposite was observed for HS QTL. The proportion of CB QTL decreased as FD deviated from 0·5. Accuracy of map position tended to be greatest for CB QTL. Models were applied to a cross of Berkshire and Yorkshire pig breeds and revealed 160 (40) QTL at the 5% chromosome (genome)-wise level for the 39 growth, carcass composition and quality traits, of which 72, 54, and 34 were declared as LC, HS and CB QTL. Fourteen CB QTL were detected only by the combined model. Thus, the combined model can increase power to detect QTL and mapping accuracy and enable characterization of QTL that segregate within breeds.

Objectives: To investigate possible obesity candidate genes in regions of porcine quantitative trait loci (QTL) for fat deposition and obesity‐related phenotypes. Research Methods and Procedures: Chromosome mapping and QTL analyses of obesity candidate genes were performed using DNA panels from a reference pig family. Statistical association analyses of these genes were performed for fat deposition phenotypes in several other commercial pig populations. Results: Eight candidate genes were mapped to QTL regions of pig chromosomes in this study. These candidate genes also served as anchor loci to determine homologous human chromosomal locations of pig fat deposition QTL. Preliminary analyses of relationships among polymorphisms of individual candidate genes and a variety of phenotypic measurements in a large number of pigs were performed. On the basis of available data, gene‐gene interactions were also studied. Discussion: Comparative analysis of obesity‐related genes in the pig is not only important for development of marker‐assisted selection on growth and fat deposition traits in the pig but also provides for an understanding of their genetic roles in the development of human obesity.

The past decade has yielded new tools for pig geneticists and breeders thanks to the considerable developments resulting from efforts to map the pig genome. The pig genetic linkage map now has nearly 5000 loci including several hundred genes, microsatellites and amplified fragment length polymorphisms (AFLP) markers. Using tools that include somatic cell hybrid panels and radiation hybrid panels, the physical genetic map is also growing rapidly and has over 4000 genes and markers. Scientists using both exotic and commercial breeds for quantitative trait loci (QTL) scans and candidate gene analyses have identified a number of important chromosomal regions and individual genes associated with growth rate, leanness, feed intake, meat quality, litter size and disease resistance. Using marker-assisted selection (MAS) the commercial pig industry is actively incorporating these gene markers and traditional performance information to improve traits of economic importance in pig production. Researchers now have novel tools including pig gene arrays and advanced bioinformatics that are being exploited to find new candidate genes and to advance the understanding of gene function in the pig. Sequencing of the pig genome has been initiated and further sequencing is now being considered. Advances in pig genomics and directions for future research and the implications to both the pig industry and human health are reviewed.

Analysis of the genetic variation of an endangered population is an important component for the success of conservation. Animals from two local Romanian pig breeds, the Mangalitsa and Bazna, were analysed for variation at a number of genetic loci using PCR-based DNA tests. Polymorphism was assessed at loci which 1) are known to cause phenotypic variation, 2) are potentially involved in trait differences or 3) are putative candidate genes. The traits considered are disease resistance, growth, coat colour, meat quality and prolificacy. Even though the populations are small and the markers are limited to specific genes, we found significant differences in five of the ten characterised loci. In some cases the observed allele frequencies were interesting in relation to gene function and the phenotype of the breed. These breeds are part of a conservation programme in Romania and marker information may be useful in preserving a representative gene pool in the populations. The use of polymorphisms in type 1 (gene) markers may be a useful complement to analysis based on anonymous markers.

The accurate estimation of the probability of identity by descent (IBD) at loci or genome positions of interest is paramount to the genetic study of quantitative and disease resistance traits. We present a Monte Carlo Markov Chain method to compute IBD probabilities between individuals conditional on DNA markers and on pedigree information. The IBDs can be obtained in a completely general pedigree at any genome position of interest, and all marker and pedigree information available is used. The method can be split into two steps at each iteration. First, phases are sampled using current genotypic configurations of relatives and second, crossover events are simulated conditional on phases. Internal track is kept of all founder origins and crossovers such that the IBD probabilities averaged over replicates are rapidly obtained. We illustrate the method with some examples. First, we show that all pedigree information should be used to obtain line origin probabilities in F2 crosses. Second, the distribution of genetic relationships between half and full sibs is analysed in both simulated data and in real data from an F2 cross in pigs.

In this study, the tail muscle microbiota of pacific white shrimp (Litopenaeus vannamei) sourced from five countries across Central and South America and Southeast Asia were determined and compared. The genomic DNA was sequenced at around 10× coverage for each geographical location and was assembled de novofor comparative analysis. The assembled sequences for all the lines were classified based on their similarity to the sequences in the public database. We found that there is high correlation among the microbiota of shrimp from disparate regions, as well as the presence of some DNA from bacteria known to cause food poisoning in humans. Sequencing data has been deposited at NCBI-SRA database and can be found under the BioProject ID PRJNA282154.

Abstract.: Eleven genes were mapped to the porcine genome with the aim of improving the human-porcine comparative gene map. Five of these genes were from regions of the human genome painted by porcine chromosomal probes; of these, two mapped to chromosomes not expected from the painting results. Among the six genes from human regions not painted by porcine chromosomal probes, three genes did not map where expected by the principle of parsimony. Several of the gene assignments indicate the existence of small regions of conserved synteny not detected by heterologous chromosome painting, especially in telomeric regions. We have also detected new rearrangements in gene order within the regions of correspondence between human Chromosome (HSA) 15 and porcine Chromosome (SSC) 1 as well as between HSA4 and SSC8.

The overall order of the regions of the swine major histocompatibility complex (MHC), the SLA complex, was determined by pulsed field gel electrophoresis (PFGE). It was found that the order of the regions is class II-class III-class I. A class I probe hybridized to a 420 kbMlu I and a 420 kbNot I fragment as did a class III probe forC2. None of the class II probes hybridized to these fragments. Thus, linkage of class I to class III was shown. The class IIiC2, Bf, andC4 genes were found to reside in a 190 kbNot I fragment. Linkage of class III and class II genes was shown when both the class IIiC4 and the class IiDR probes hybridized to the same 195 kbSac II and 340 kbNot I fragments. The class I probe did not hybridize to these fragments. The order of the regions, class II-class III-class I, is similar to that of human MHC genes and may have been conserved in evolution so that coordinated expression of MHC genes could be achieved.

Complement regulatory proteins have become important targets to potentially modulate inflammatory reactions or transplant rejection. Since pig into human xenotransplantation could potentially overcome the enormous shortage of donor organs and tissues, characterization of porcine complement regulatory proteins is critical.

Porcine periweaning failure to thrive syndrome (PFTS) can be defined by anorexia, lethargy, progressive debilitation and compulsive behaviours that occur in seemingly healthy pigs within two to threeweeks of weaning in the absence of any known infectious, nutritional, management or environmental factors. A genetic component has been hypothesised for this syndrome. In the present study, 119 commercial pigs (80 cases and 39 controls) were genotyped with the porcine 80K single nucleotide polymorphism‐chip and were analysed with logistic regression and two Fixation Index‐based approaches. The analyses revealed several regions on chromosomes 1, 3, 6 and 11 with moderate divergence between cases and controls, particularly three haplotypes on SSC3 and 11. The gene‐based analyses of the candidate regions revealed the presence of genes that have been reported to be associated with phenotypes like PFST including depression (PDE10A) and intestinal villous atrophy (CUL4A). It is important to increase the effort of collecting more samples to improve the power of these analyses.

Small RNA represent several unique noncoding RNA classes that have important function in the development of germ cells and early embryonic development. Deep sequencing was performed on small RNA from cumulus cells (recovered from germinal vesicle [GV] and metaphase II-arrested [MII] oocytes), GV and MII oocytes, in vitro fertilization-derived embryos at 60 h postfertilization (4- to 8-cell stage), and Day 6 blastocysts. Additionally, a heterologous miRNA microarray method was also used to identify miRNA expressed in the oocyte during in vitro maturation. Similar to the results of expression analysis of other species, these data demonstrate dynamic expression regulation of multiple classes of noncoding RNA during oocyte maturation and development to the blastocyst stage. Mapping small RNA to the pig genome indicates dynamic distribution of small RNA organization across the genome. Additionally, a cluster of miRNA and Piwi-interacting RNA (piRNA) was discovered on chromosome 6. Many of the small RNA mapped to annotated repetitive elements in the pig genome, of which the SINE/tRNA-Glu and LINE/L1 elements represented a large proportion. Two piRNA (piR84651 and piR16993) and seven miRNA (MIR574, MIR24, LET7E, MIR23B, MIR30D, MIR320, and MIR30C) were further characterized using quantitative RT-PCR. Secretory carrier membrane protein 4 (SCAMP4) was predicted to be subject to posttranscriptional gene regulation mediated by small RNA, by annotating small RNA reads mapped to exonic regions in the pig genome. Consistent with the prediction results, SCAMP4 was further confirmed to be differentially expressed at both transcriptional and translational levels. These data establish a small RNA expression profile of the pig cumulus-oocyte complex and early embryos and demonstrate their potential capacity to be utilized for predictions of functional posttranscriptional regulatory events.

In eukaryotes, small non-coding RNA capable of binding to Argonaute (Ago) proteins include microRNA (miRNA), small interfering RNA (siRNA) and 24-30 nucleotide long Piwi-interacting RNA (piRNA). Both miRNA and siRNA, which are produced from long double stranded RNA (dsRNA) precursors by Dicer, associate with the Ago subclade members of the Argonaute protein family and impact posttranscriptional gene regulation through sequence-specific interactions with mRNA molecules. Originally discovered in Drosophila, piRNA belong to the Piwi subclade members of the Argonautes and play essential roles in genome protection against retrotransposon inversion through silencing and gene regulation by targeting protein-coding genes during germ cell development. Although piRNA have been identified in Drosophila, zebrafish and mice, there is little information regarding piRNA expression in domestic animals. Objective of this study was to identify piRNA expression in pig cumulus oocyte complexes (COC) and during early embryo development using high-throughput deep sequencing. Sow ovaries were obtained from a local abattoir and COC were aspirated from 3 to 6-mm follicles. COC were in vitro matured for 42-44 hr followed by in vitro fertilization and culture for 6 days to the blastocyst stage of development. Total RNA was extracted from cumulus cells of germinal vesicle stage (GV) and MII arrested oocytes, GV oocytes (n=1245), MII arrested oocytes (n=1309), 4-8 cell in vitro fertilized embryos (n=787) and day 6 blastocysts (n=395). Small RNA libraries were created using the Small RNA Expression Kit (Ambion) and subjected to deep sequencing using the SOLiD sequencing system (Applied Biosystems). De novo piRNAs in the pig genome were predicted using piRNA version 2 datasets downloaded from a comprehensive mammalian noncoding RNA database (RNAdb) to compare against the pig genome using Megablast as many piRNA are conserved across species. Short sequence reads were aligned back to the pig genome to profile the abundance of piRNA with at least 3 reads from the small RNA library. We found 44,278 piRNA reads corresponding to 71 different genome locations in GV oocytes, 31,311 reads corresponding to 71 locations in MII oocytes, 30,607 reads corresponding to 72 locations in 4-8 cell embryos, 19,321 reads corresponding to 70 locations in blastocysts. In cumulus cells, 48,569 reads corresponding to 84 locations in cumulus from GV oocytes and 30,574 reads corresponding to 82 locations in cumulus cells from MII arrested oocytes. One piRNA, on chromosome 6 (SSC6: 246415-246443), was abundantly expressed in oocytes and embryos, representing 41.6% to 80.7% of the total piRNA reads depending on the stage of development. These data demonstrate piRNA expression in pig oocytes, embryos and cumulus cells, and provide important information for future studies to determine the potential involvement of specific piRNA in female germ cell development in pigs. This project was supported by National Research Initiative Competitive Grant no. 2008-35205-05309 and 2008-35205-18712 from the USDA National Institute of Food and Agriculture.(poster)

Country hams are dry-cured products from the Southeastern region of the USA. This high value product requires quality fresh meat to avoid later processing problems. The marker SREBF1is a transcription factor involved in the regulation of fatty acid synthesis and anti-oxidative enzyme transcription. The SREBF1gene and its regulators, SCAPand MBTPS1, were investigated for associations with several meat quality traits in country hams. After single nucleotide polymorphism (SNP) identification, PCR-RFLP tests were designed for one polymorphism in each of the three investigated genes. Meat quality and physical traits were collected on 299 fresh hams. Significant associations were found with ham yield (MBTPS1, P<0.05 and SREBF1×MBTPS1, P<0.05) and ham circumference (MBTPS1, P<0.05 and SREBF1×MBTPS1, P<0.01), Hunter A colour score on fresh meat (SREBF1, P<0.05), Hunter B colour score on cured meat (MBTPS1, P<0.05 and SREBF1×MBTPS1, P<0.01), moisture (SCAP×MBTPS1, P<0.05) and salt percentage (SREBF1×SCAP, P<0.05). Our findings provide initial evidence that SNPs in SREBF1, SCAPand MBTPS1are associated with some country ham quality traits. Breeders could use these gene tests to improve their animals, which would in turn improve country ham processing and other desired production goals.

The accurate estimation of the probability of identity by descent (IBD) at loci or genome positions of interest is paramount to the genetic study of quantitative and disease resistance traits. We present a Monte Carlo Markov Chain method to compute IBD probabilities between individuals conditional on DNA markers and on pedigree information. The IBDs can be obtained in a completely general pedigree at any genome position of interest, and all marker and pedigree information available is used. The method can be split into two steps at each iteration. First, phases are sampled using current genotypic configurations of relatives and second, crossover events are simulated conditional on phases. Internal track is kept of all founder origins and crossovers such that the IBD probabilities averaged over replicates are rapidly obtained. We illustrate the method with some examples. First, we show that all pedigree information should be used to obtain line origin probabilities in F2 crosses. Second, the distribution of genetic relationships between half and full sibs is analysed in both simulated data and in real data from an F2 cross in pigs.