A genome-wide association study for gestation length in swine.

Selection for increased litter size in swine has potentially resulted in a correlated increase in preweaning mortality. Additional selection criteria should be considered when selecting for increased litter size to account for associated decreases in piglet quality, specifically piglet survival, initial weight and growth. Traits such as gestation length (GL), which have been associated with piglet performance, could be utilized to improve piglet development and survivability. The objective of this study was to conduct a genome-wide association study to identify genomic regions associated with GL in differing parities in swine (n = 831) from the University of Nebraska-Lincoln reproductive longevity project. Gestation length was calculated as the number of days between last insemination administered and farrowing. Sows were genotyped with the Illumina SNP60 BeadArray, and the data were analyzed using Bayesian mixture models for GL at parity 1, 2, 3 and 4 (GL1, GL2, GL3 and GL4 respectively). Means (SD) for GL1-GL4 were 113 (1.4), 114 (1.2), 114 (1.3) and 115 (1.2) respectively. Posterior mean heritability estimates (PSD) for GL1, GL2, GL3 and GL4 were 0.33 (0.06), 0.34 (0.07), 0.32 (0.08) and 0.20 (0.08) respectively. Rank correlations between genomic estimated breeding values between GL1 and GL2, GL3 and GL4 respectively were moderate: 0.67, 0.65 and 0.60. The top SNP (ASGA0017859, SSC4, 7.8 Mb), located in the top common genomic region associated with GL1, GL2 and GL3, was associated with a difference of 1.1 days in GL1 between homozygote genotypes (P < 0.0001). The results of this study suggest that GL is a largely polygenic trait with relatively minor contributions from multiple genomic regions.

Genomic investigation of piglet resilience following porcine epidemic diarrhea outbreaks.

Porcine epidemic diarrhea virus (PEDV) belongs to the Coronaviridae family and causes malabsorptive watery diarrhea, vomiting, dehydration and imbalanced blood electrolytes in pigs. Since the 1970s, PED outbreaks have become a source of problems in pig producing countries all over the world, causing large economic losses for pig producers. Although the infection in adults is not fatal, in naïve suckling piglets mortality is close to 100%. In this study, we investigated genome-wide differences between dead and recovered suckling piglets from commercial farms after PED outbreaks. Samples from 262 animals (156 dead and 106 recovered) belonging to several commercial lines were collected from five different farms in three different countries (USA, Canada and Germany) and genotyped with the porcine 80K SNP chip. Mean Fst value was calculated in 1-Mb non-overlapping windows between dead and recovered individuals, and the results were normalized to find differences within the comparison. Seven windows with high divergence between dead and recovered were detected-five on chromosome 2, one on chromosome 4 and one on chromosome 15-in total encompassing 152 genes. Several of these genes are either under- or overexpressed in many virus infections, including Coronaviridae (such as SARS-CoV). A total of 32 genes are included in one or more Gene Ontology terms that can be related to PED development, such as Golgi apparatus, as well as mechanisms generally linked to resilience or diarrhea development (cell proliferation, ion transport, ATPase activity). Taken together this information provides a first genomic picture of PEDV resilience in suckling piglets.

Genetic associations for gilt growth, compositional, and structural soundness traits with sow longevity and lifetime reproductive performance.

The objective of this study was to estimate genetic associations for gilt growth, compositional, and structural soundness with sow longevity and lifetime reproduction. Performance and pedigree information from 1,447 commercial females from 2 genetic lines were included in the data analyzed. Growth was expressed as days to 113.5 kg BW (DAYS) and compositional traits included loin muscle area (LMA), 10th rib backfat (BF10), and last rib backfat (LRF). Structural soundness traits included body structure traits [length (BL), depth (BD), width (BWD), rib shape (BRS), top line (BTL), and hip structure (BHS)], leg structure traits [front legs: legs turned (FLT), buck knees (FBK), pastern posture (FPP), foot size (FFS), and uneven toes (FUT); rear legs: legs turned (RLT), leg posture (RLP), pastern posture (RPP), foot size (RFS), and uneven toes (RUT)], and overall leg action (OLA). Lifetime (LT) and removal parity (RP) were considered as longevity traits whereas lifetime reproductive traits included lifetime total number born (LNB), lifetime number born alive (LBA), number born alive per lifetime day (LBA/LT), and percentage productive days from total herd days (PD%). Genetic parameters were estimated with linear animal models using the average information REML algorithm. Second, to account for censored longevity and lifetime reproduction records, genetic parameters were estimated using Markov Chain Monte Carlo and Gibbs sampling methods. Similar estimates were obtained across the analysis methods. Heritability estimates for growth and compositional traits ranged from 0.50 to 0.70 and for structural soundness traits from 0.07 to 0.31. Longevity and lifetime reproductive trait heritability estimates ranged from 0.14 to 0.17 when REML was used. Unfavorable genetic correlations were obtained for DAYS with LT, RP, LNB, LBA, and PD% and for LRF with PD%. However, LMA was favorably associated with LT, RP, and LNB. Moderate to high correlations were obtained for BL and BRS with all longevity and lifetime reproductive traits. Correlations of BWD with LT and RP were moderate. Associations for leg soundness traits with longevity and lifetime reproductive traits were mainly low and nonsignificant (P ≥ 0.10). However, RLP was moderately correlated with LBA/LT and PD%. Current results indicate that selection for fewer DAYS has an antagonistic effect on lifetime performance. Furthermore, great BL, flat BRS, narrow BWD, and upright RLP seem detrimental to sow longevity and lifetime reproduction.

Genetic parameters for growth, body composition, and structural soundness traits in commercial gilts.

The objective of this study was to estimate genetic parameters for growth, body composition, and structural soundness traits in commercial gilt lines. The data included 1,449 gilts: 462 females from a grandparent maternal line and 987 from a parent maternal line. Growth was expressed as number of days to a constant 113.5 kg BW (DAYS) and compositional traits included loin muscle area (LMA), 10th rib backfat (BF10), and last rib backfat (LRF). Subjective structural soundness evaluation was completed using a 9-point scale and included: body length (BL), body depth (BD), body width (BWD), rib shape (BRS), top line (BTL), and hip structure (BHS); front legs: legs turned (FLT), buck knees (FBK), pastern posture (FPP), foot size (FFS), and uneven toes (FUT); rear legs: legs turned (RLT), leg posture (RLP), pastern posture (RPP), foot size (RFS), and uneven toes (RUT); and overall leg action (OLA). Genetic parameters were estimated with multivariate linear animal models, using the average information REML algorithm. Heritability estimates for growth and body composition traits ranged from 0.50 to 0.70, for body structure traits from 0.15 to 0.31, for leg structure traits from 0.07 to 0.31, and the estimate for OLA was 0.12. Several moderate to high genetic correlations were obtained among body structure traits, whereas correlations among leg structure traits were mainly low and nonsignificant. A strong correlation was found between FPP and OLA (P < 0.001); more upright FPP coincided with inferior OLA. Furthermore, FBK and FFS appeared to be favorably associated with OLA (0.05 < P < 0.10). Body structure trait correlations among each other and with leg soundness traits were primarily favorable. Correlations indicated that great BL and high BTL coincided with each other and deterioration of other structural soundness traits. Although genetic correlations obtained for DAYS and backfat measurements with structural soundness traits had an unfavorable trend, they were mainly low to moderate (i.e., simultaneous genetic improvement would be possible, including adversely associated traits). Due to greater heritabilities, faster genetic change could be expected for compositional and body structure traits than leg structure traits. Because of the genetic relationship among the trait groups, using information across traits when making selection decisions could result in genetic improvement among leg soundness traits.

Evaluation of enrichment media for improved PCR-based detection of V. cholerae and V. vulnificus from estuarine water and plankton.

AIMS: Pathogenic Vibrio spp., including V. cholerae and V. vulnificus, are commonly found along the estuaries of the south-east United States; however, it is often difficult to recover these species directly from environmental samples. Pre-enrichment assays are commonly used to improve the detection of pathogenic vibrios from environmental sources. Here, we evaluated a novel enrichment procedure using freshly collected and autoclaved natural estuarine water amended with 1% peptone (designated as estuarine peptone water, EPW) and compared it to traditional alkaline peptone water (APW) for detection by PCR of V. cholerae and V. vulnificus. METHODS AND RESULTS: Of the 50 samples collected in total, V. cholerae DNA was detected in APW 10% of the time and in EPW 40% of the time. Likewise, the cholera toxin gene (ctxA) was detected in 4 vs 18% of the samples using APW and EPW, respectively. Conversely, APW showed improved recovery for V. vulnificus relative to EPW with respective detection frequencies of 46 and 20%. Results showed similar patterns across different sample types (water and plankton). CONCLUSIONS: While enrichment in traditional APW was adequate for the recovery of Vibrio vulnificius, use of sterile estuarine water amended with peptone significantly improved the detection of V. cholerae and the virulence gene ctxA from estuarine sources.

Effect of polymorphisms in candidate genes on reproduction traits in Finnish pig populations.

Reproduction traits play an important role in economically viable piglet production and are closely related to the quality and length of the productive life of the sow. A increased removal rate of young sows is undesirable not only because of the associated financial penalties incurred, but also because of ethical concerns. Candidate genes and gene pathways have been identified for fertility in model species, and recent studies have provided evidence that polymorphisms within these genes are associated with reproduction traits in American Yorkshire/Large White and Landrace populations. In this study we evaluated the impact of single polymorphisms (n = 7) in 7 candidate genes on reproductive efficiency in Finnish Yorkshire (n = 280) and Landrace (n = 271) populations: IGFBP1, IGFBP2, IGFBP3, IGFBP5, CPTIA (carnitine O-palmitoyltransferase I), COX2 (PG-endoperoxide synthase 2, also known as cyclooxygenase-2), and SLC22A5 [organic cation/carnitine transporter 2 (solute carrier family member I), OCTN2]. In the Finnish Yorkshire population, only 4 of the analyzed markers were polymorphic. Significant effects on farrowing time were detected from the Yorkshire data, with polymorphisms within the genes CPT1A [a (allele substitution effect of allele A) = 2.97 d for age at first farrowing)], IGFBP3 (a = 0.54 d for farrowing interval of parities >1), and IGFBP5 (a = 3.22, 1.27, and 0.85 d for age at first farrowing and farrowing interval in the first and later parities, respectively). For the Landrace population, 6 markers were polymorphic, and significant effects were detected for traits affecting litter size. The polymorphism within the COX2 gene had an additive effect of 0.3 piglets for litter size in parities >1, and the IGFBP1 gene had an additive effect of 0.21, 0.26, and 0.11 piglets for litter size in the first parity, parities >1, and stillborn in parities >1, respectively. The additive effect of the SNP within the IGFBP2 gene was 0.16, 0.09, and 0.09 piglets for litter size in parities >1 and stillborn in the first and later parities, respectively. Finally, the IGFBP5 gene had an additive effect of 0.18, 0.07, and 0.07 piglets for litter size in the first parity, stillborn in parities >1, and mortality between farrowing and weaning in the first parity, respectively. These results support the suitability of the candidate gene approach for identification of markers to improve the reproductive performance of sows and to provide potential markers for marker-assisted selection.

An evaluation of four candidate genes for use in selection programmes aimed at increased intramuscular fat in Duroc swine.

A sufficient level of intramuscular fat (IMF) is needed to enhance consumer acceptance of pork products, and is currently receiving greater attention within swine genetic improvement programmes. An examination of previously described and novel genetic variants within candidate genes for IMF deposition was performed to evaluate potential use of genetic markers in marker-assisted selection (MAS). Biological candidate genes implicated to play a role in adipogenesis were investigated within two different lines of purebred Duroc pigs. These included MC4R, FABP3, DLK1, and TCF7L2. Significant variation in IMF within the control line was described by the MC4R genotype and a novel BsrfI single nucleotide polymorphism within the FABP3 gene. Genetic markers for DLK1 and TCF7L2 evaluated in this population are not currently recommended for selection in Duroc swine. Existence of MC4R and FABP3 mutations may be useful markers in MAS aimed at IMF improvement, provided that gene effects are segregating and the presence of an association is detected within the population. However, additional work to confirm the use of the investigated genetic markers in selection programmes is needed.

Identification of genetic markers for productive life in commercial sows.

Escalating replacement rates and production costs warrant attention on sow productive life (SPL). Increasing average SPL by one-tenth of 1 parity would result in an annual revenue increase of over $15 million in the United States. Research in model organisms has revealed conserved genes and gene pathways that lead to longer lifespan. The most prominent gene pathways are those involved in growth, most notably genes in the IGF pathway that serve to mimic the response of caloric restriction. The objective of this research was to test the hypothesis that these well conserved genes and gene pathways could also play a role in SPL, even though the productive life of sows is both a measure of longevity and their reproductive performance. Preliminary research on 3 distinct populations of over 2,000 animals suggested that several genes were associated with components of SPL. Genetic markers were then analyzed against the corresponding records of the sows for reproductive and longevity traits using a validation population of 2,000 commercial females. Right censored data were used to test associations of genetic markers with survival to defined time points. Three distinct models of survival analysis were implemented using nonparametric estimates of the survival distribution in a sequential order, using a parametric accelerated failure time model with a Weibull distribution of the error term, and a Cox proportional hazards model, which is a semiparametric model that uses an unspecified baseline hazard function. The genetic markers CCR7 and CPT1A were significantly associated (P < 0.05) with survival using the nonparametric model and tended (P < 0.1) toward significance using the parametric and semiparametric models with significantly different effects (P < 0.05) between some genotype classes. Genetic markers for MBL2, IGFBP3, and WARS2 also tended (P < 0.1) toward significance for survival traits, but were not consistent. Mixed model analyses were used to determine the associations of these genetic markers with reproductive traits. The genetic markers for IGFBP1, MBL2, CPT1A, CCR7, SLC22A5, and ACE were significantly (P < 0.05) associated with at least 1 reproductive trait. These results show that molecular markers should be considered for use in marker-assisted selection to improve SPL.

A Limousin specific myostatin allele affects longissimus muscle area and fatty acid profiles in a Wagyu-Limousin F2 population.

A microsatellite-based genome scan of a Wagyu x Limousin F(2) cross population previously demonstrated QTL affecting LM area and fatty acid composition were present in regions near the centromere of BTA2. In this study, we used 70 SNP markers to examine the centromeric 24 megabases (Mb) of BTA2, including the Limousin-specific F94L myostatin allele (AB076403.1; 415C > A) located at approximately 6 Mb on the draft genome sequence of BTA2. A significant effect of the F94L marker was observed (F = 60.17) for LM area, which indicated that myostatin is most likely responsible for the effect. This is consistent with previous reports that the substitution of Leu for Phe at AA 94 of myostatin (caused by the 415C > A transversion) is associated with increased muscle growth. Surprisingly, several fatty acid trait QTL, which affected the amount of unsaturated fats, also mapped to or very near the myostatin marker, including the ratio of C16:1 MUFA to C16:0 saturated fat (F = 16.72), C18:1 to C18:0 (F = 18.88), and total content of MUFA (F = 17.12). In addition, QTL for extent of marbling (F = 14.73) approached significance (P = 0.05), and CLA concentration (F = 9.22) was marginally significant (P = 0.18). We also observed associations of SNP located at 16.3 Mb with KPH (F = 15.00) and for the amount of SFA (F = 12.01). These results provide insight into genetic differences between the Wagyu and Limousin breeds and may lead to a better tasting and healthier product for consumers through improved selection for lipid content of beef.

SNP discovery in genes affecting leg health traits in pigs.

Sow reproductive life is influenced by lameness issues. It has been reported that up to 44 percent of sows have locomotive problems. To date, few genome scans or association studies have been conducted to look at genes controlling lameness and other gait traits. In addition to health problems associated with leg and bone disorders, the pig has been suggested to be a good model for human bone disorders. Hence, the present study examined 134 porcine genes affecting skeletal development, mineral metabolism and other candidate genes for single nucleotide polymorphism (SNP) discovery. Atotal of over 370 SNPs have been identified to date and are being mapped. These SNPs are also being investigated for their associations with gait and locomotion problems in approximately 2,000 commercial pigs scored for various leg and locomotion traits. The association analysis of 22 genes revealed that the genes CALCR, HDBP CALCA, MTHFR, OXTR, IHH, ANKH, LRCH1 and OPN were significantly associated with leg and body conformation traits which affect the health and productivity of pigs.

Single nucleotide polymorphism identification, linkage and radiation hybrid mapping of the porcine pituitary adenylate cyclase-activating polypeptide type I receptor gene to chromosome 18.

Pituitary adenylate cyclase-activating polypeptide (PACAP) is a neuropeptide with diverse biological actions. Type I PACAP receptors (PACAPR) are specific for PACAP, whereas type II and III PACAPRs are less restricted. To localize and analyse the variation of this gene, a 559-bp long intronic fragment of the porcine PACAPR gene was amplified by polymerase chain reaction and sequenced in samples from five different pig breeds. One single nucleotide polymorphism was identified and its allele frequency was determined in all five breeds. Linkage analysis in a Berkshire x Yorkshire reference family placed the PACAPR gene on chromosome 18, between SW787 and S0062 (SW787- 8.1 cM -PACAPR- 3.0 cM -S0062). Radiation hybrid mapping confirmed that the PACAPR gene was linked to SW1682 on chromosome 18 (28.8 cR(3000); LOD = 10.4).

Cracking the genomic piggy bank: identifying secrets of the pig genome.

Though researchers are uncovering valuable information about the pig genome at unprecedented speed, the porcine genome community is barely scratching the surface as to understanding interactions of the biological code. The pig genetic linkage map has nearly 5,000 loci comprised of genes, microsatellites, and amplified fragment length polymorphism markers. Likewise, the physical map is becoming denser with nearly 6,000 markers. The long awaited sequencing efforts are providing multidimensional benefits with sequence available for comparative genomics and identifying single nucleotide polymorphisms for use in linkage and trait association studies. Scientists are using exotic and commercial breeds for quantitative trait loci scans. Additionally, candidate gene studies continue to identify chromosomal regions or genes associated with economically important traits such as growth rate, leanness, feed intake, meat quality, litter size, and disease resistance. The commercial pig industry is actively incorporating these markers in marker-assisted selection along with traditional performance information to improve said traits. Researchers are utilizing novel tools including pig microarrays along with advanced bioinformatics to identify new candidate genes, understand gene function, and piece together gene networks involved in important biological processes. Advances in pig genomics and implications to the pork industry as well as human health are reviewed.