Search

Early predictions of cows’ probability of survival to different lactations would help farmers in making successful management and breeding decisions. For this purpose, this research explored the adoption of joint models for longitudinal and survival data in the dairy field. An algorithm jointly modelled daily first-lactation sensor data (milk yield, body weight, rumination time) and survival data (i.e., time to culling) from 6 Holstein dairy farms. The algorithm was set to predict survival to the beginning of the second and third lactations (i.e., second and third calving) from sensor observations of the first 60, 150, and 240 days in milk of cows’ first lactation. Using 3-time-repeated 3-fold cross-validation, the performance was evaluated in terms of Area Under the Curve and expected error of prediction. Across the different scenarios and farms, the former varied between 45% and 76%, while the latter was between 3.5% and 26%. Significant results were obtained in terms of expected error of prediction, meaning that the method provided survival probabilities in line with the observed events in the datasets (i.e., culling). Furthermore, the performances were stable among farms. These features may justify further research on the use of joint models to predict the survival of dairy cattle.

[Background]: The comparison of expression QTL (eQTL) maps obtained in different tissues is an essential step to understand how gene expression is genetically regulated in a context-dependent manner. In the current work, we have compared the transcriptomic and eQTL profiles of two porcine tissues (skeletal muscle and liver) which typically show highly divergent expression profiles, in 103 Duroc pigs genotyped with the Porcine SNP60 BeadChip (Illumina) and with available microarray-based measurements of hepatic and muscle mRNA levels. Since structural variation could have effects on gene expression, we have also investigated the co-localization of cis-eQTLs with copy number variant regions (CNVR) segregating in this Duroc population., [Results]: The analysis of differential expresssion revealed the existence of 1204 and 1490 probes that were overexpressed and underexpressed in the gluteus medius muscle when compared to liver, respectively (|fold-change| > 1.5, q-value < 0.05). By performing genome scans in 103 Duroc pigs with available expression and genotypic data, we identified 76 and 28 genome-wide significant cis-eQTLs regulating gene expression in the gluteus medius muscle and liver, respectively. Twelve of these cis-eQTLs were shared by both tissues (i.e. 42.8% of the cis-eQTLs identified in the liver were replicated in the gluteus medius muscle). These results are consistent with previous studies performed in humans, where 50% of eQTLs were shared across tissues. Moreover, we have identified 41 CNVRs in a set of 350 pigs from the same Duroc population, which had been genotyped with the Porcine SNP60 BeadChip by using the PennCNV and GADA softwares, but only a small proportion of these CNVRs co-localized with the cis-eQTL signals., [Conclusion]: Despite the fact that there are considerable differences in the gene expression patterns of the porcine liver and skeletal muscle, we have identified a substantial proportion of common cis-eQTLs regulating gene expression in both tissues. Several of these cis-eQTLs influence the mRNA levels of genes with important roles in meat (CTSF) and carcass quality (TAPT1), lipid metabolism (TMEM97) and obesity (MARC2), thus evidencing the practical importance of dissecting the genetic mechanisms involved in their expression.

We aimed to analyse the genetic diversity of Romanian wild boars and to compare it with that from other wild boar and pig populations from Europe and Asia. Partial sequencing of the mitochondrial encoded cytochrome b (MT-CYB) gene from 36 Romanian wild boars and 36 domestic pigs (Mangalitza, Bazna and Vietnamese breeds) showed that the diversity of Romanian wild boars and Mangalitza pigs is fairly reduced and that most of the members of these two populations share a common MT-CYB haplotype. Besides, in strong contrast with the Bazna animals, Romanian wild boars and Mangalitza swine did not carry Asian variants at the MT-CYB locus. The autosomal genotyping of 18 Romanian wild boars with the Illumina Porcine SNP60 BeadChip revealed that their genetic background is fundamentally European, even though signs of a potential Near Eastern ancestry (~25%) were detectable at K = 4 (the most significant number of clusters), but not at higher K-values. Admixture analysis also showed that two wild boars are of a hybrid origin, which could be explained by the mating of feral animals with domestic pigs. Finally, a number of Romanian wild boars displayed long runs of homozygosity, an observation that is consistent with the occurrence of past population bottlenecks and the raise of inbreeding possibly due to overhunting or to the outbreak of infectious diseases.

In humans, the variability of the melanocortin 1 receptor (MC1R) gene has been associated with geography, being mainly determined by the amount of exposure to sunlight. Studies performed in pigs have also evidenced the existence of a geographic component in the distribution of MC1R haplotypes, probably as a consequence of an ancient split between Asian and European wild boars. Herewith, we have partially resequenced the caprine MC1R coding region in 58 goats from distinct geographic locations i.e. Colombia, Italy, Spain, France, Greece, Romania, Iran and Africa. The resulting dataset was merged with 39 previously published caprine MC1R sequences and a median joining network was built. This phylogenetic analysis did not yield any evidence of a relationship between geography and the clustering of caprine MC1R sequences, a result that was confirmed by performing a Mantel test with a previously published dataset of nine goat breeds (N = 319) with available MC1R genotypes. The majority of caprine MC1R variation was non-synonymous (c.676A > G, c.748G > T, c.764G > A and c.801C > G) and predicted to have functional effects. An analysis of goat MC1R sequences with the PAML 4 software provided evidence that two SNPs (c.764G > A and c.801C > G) might evolve under positive selection. The apparent lack of any link between caprine MC1R variation and geography might be explained by a complex array of factors including artificial selection for pigmentation phenotypes and recent divergence amongst goat breeds.

The adiponectin (ADIPOQ) locus is a positional and functional candidate gene for 2 porcine chromosome 13 (SSC13) QTL influencing cholesterol (CHOL) and low-density lipoprotein (LDL) concentrations in 190-d-old pigs. By sequencing 2.37 kb of the pig ADIPOQ cDNA, we have identified 1 c.*1512G > T 3′ untranslated region polymorphism that has been genotyped in a Duroc pig commercial population with records for serum lipid levels at 45 and 190 d of age. Statistical analysis of the data have revealed significant associations between the ADIPOQ genotype and CHOL (P = 0.0040) and LDL (P = 0.0011) concentrations at 190 d but not at 45 d. In family 3, most of the SSC13 QTL effects on LDL levels at 190 d were explained by the ADIPOQ genotype. We also found an association with triglyceride levels at 45 d (P = 0.0060) but not at 190 d. Measurement of allelic mRNA imbalance demonstrated that the G and T alleles are expressed at very similar levels in muscle and fat tissues, indicating that the c.*1512G > T polymorphism does not affect transcript abundance. As a whole, results obtained in the current work as well as previous data gathered in humans and pigs provide evidence that the magnitude of associations between blood lipid phenotypes and candidate loci genotypes may vary depending on the age of the individual, therefore suggesting the existence of dynamic genotype × environment interactions changing on a temporal scale.

The CD36 molecule plays a key role in the uptake of long-chain fatty acids. In a previous study, we demonstrated that the CD36 gene is duplicated in goats. Moreover, both copies (CD36 and CD36-like) display highly divergent mRNA expression profiles. Herewith, we have analyzed whether four polymorphisms mapping to CD36 (c.394A > G, c.*141C > T and c.*427T > A) and CD36-like (c.390A > C) genes are associated with milk yield and composition. Murciano-Granadina goats with records for dairy traits (N = 309, 1005 registers) and milk fatty acid composition phenotypes (N = 176, 490 registers) were genotyped for these four markers and association analyses were carried out. We found a highly significant association between c.*427T > A CD36 polymorphism and milk palmitoleic content (P < 0.0001). Besides, the c.*141C > T CD36 SNP also showed a suggestive association (P = 0.04) with palmitoleic. These findings are consistent with previous studies showing that CD36 inactivation in adipocytes and neurons involves a decrease in palmitoleic content, thus suggesting a relevant role of this molecule in the specific uptake of this fatty acid. The remaining associations found (CD36 with polyunsaturated fatty acids and CD36-like with oleic and several saturated fatty acids) were significant at the nominal level but not after Bonferroni correction. These results, combined with previously reported expression data, reinforce the interest of investigating the lipid binding properties of CD36 and CD36-like through functional approaches.

The CD36 molecule plays a pivotal role in a variety of immunological and cellular processes, including pathogen recognition, inflammation and apoptosis. Herein, we demonstrate that this gene is duplicated in goats, with two copies (CD36 and CD36-like) that display highly divergent mRNA expression profiles. In this way, CD36 mRNA is mostly expressed in the adipose tissue and heart whilst CD36-like mRNA shows a high expression in the liver. We have also found evidence of the presence of two paralogous CD36 and CD36-like genes in the bovine genome, suggesting that CD36 duplication took place before goat–cattle radiation, i.e. at least 20 MYR ago. Finally, we have characterized the polymorphism of the coding regions of the goat CD36 and CD36-like genes. In doing so, we have identified one synonymous polymorphism at the CD36-like gene (c.390A>C) that displays a significant association (P = 0.04) with milk somatic cell count, a parameter often used to diagnose mastitis in domestic ruminants.

The performance of a genome scan for serum lipid traits at 45 and 190 d in 5 half-sib families of Duroc pigs allowed us to detect several pig chromosomal regions with significant effects on these phenotypes. In the current work, we aimed to refine the position of 1 chromosome 4 (SSC4) genome-wide significant QTL for serum triglyceride concentration at 190 d. Genotyping of 4 additional microsatellites allowed reduction of the 90% confidence interval of this QTL to the genomic interval between markers SW2409 and SW839. Sequencing experiments were performed to characterize the variability of 2 lipid-related genes, the lipoprotein receptor-related protein 12 (LRP12) and tribbles homolog 1 (TRIB1) loci, that map to this region. In this way, 2 (c.771A > G and c.1101A > G) and 1 (c.*156_157del) polymorphisms were identified at the LRP12 coding region and TRIB1 3′ untranslated region, respectively. Association analyses between LRP12 and TRIB1 genotypes did not reveal any significant effect on serum lipid concentrations, suggesting that variation of these two loci does not explain the segregation of the SSC4 QTL. However, highly significant associations were observed for gluteus medius saturated fatty acid content (LRP12 c.1101A > G, P = 0.0006; TRIB1 c.*156_157del, P = 0.0003). In the light of these and other findings, the potential involvement of LRP12 and TRIB1 in muscle lipid metabolism deserves to be further explored.

Solute carrier family 27A (SLC27A) molecules play a fundamental role in the cellular uptake of long-chain fatty acids in a wide variety of tissues and organs. Although polymorphisms at the pig SLC27A1 and SLC27A4 genes have been associated with backfat thickness in pigs, the existence of associations with lipid composition traits has not been investigated yet. Herewith, we have sequenced the coding regions of the SLC27A1, SLC27A2 and SLC27A4 genes in 15 Duroc pigs, revealing the existence of three single nucleotide polymorphisms (SNP) at SLC27A1 (c.441C>T, c.747A>G and c.⁎400C>T) and two additional SNP at SLC27A4 (c.243T>G and c.837C>T). The performance of an association analysis with a wide array of serum and muscle lipid traits revealed the existence of suggestive associations (nominal P-value <0.05, but not significant after correction for multiple testing) between SLC27A1 genotype and gluteus medius saturated fatty acid (SFA) and stearic contents (%), and between SLC27A4 genotype and gluteus medius SFA content (%). Similarly, SLC27A1 genotype was suggestively associated with SFA content (%) at the longisimus thoracis et lumborum muscle. These findings agree from a functional point of view with the essential role of SLC27A family members in cellular fatty acid uptake and reinforce the need of further exploring the role of these molecules in defining muscle lipid composition.

The lack of a Near Eastern genetic signature in modern European porcine breeds indicates that, although domestic pigs from the Fertile Crescent entered Europe during the Neolithic, they were completely replaced by their European counterparts in a short window of time. Whilst the absence of such genetic signature has been convincingly demonstrated at the mitochondrial level, variation at the autosomal genomes of European and Near Eastern Sus scrofa has not been compared yet. Herewith, we have explored the genetic relationships among 43 wild boar from Europe (N = 21), Near East (N = 19) and Korea (N = 3), and 40 Iberian (N = 16), Canarian (N = 4) and Mangalitza (N = 20) pigs by using a high throughput SNP genotyping platform. After data filtering, 37,167 autosomal SNPs were used to perform population genetics analyses. A multidimensional scaling plot based on genome-wide identity-by-state pairwise distances inferred with PLINK showed that Near Eastern and European wild boar populations are genetically differentiated. Maximum likelihood trees built with TreeMix supported this conclusion i.e. an early population split between Near Eastern and European Sus scrofa was observed. Moreover, analysis of the data with Structure evidenced that the sampled Iberian, Canarian and Mangalitza pigs did not carry any autosomal signature compatible with a Near Eastern ancestry, a finding that agrees well with previous mitochondrial studies. © 2013 Manunza et al.

[EN]: Sterol regulatory element binding transcription factor 1 (SREBF1) regulates the expression of genes involved in the biosynthesis of fatty acids and cholesterol. Herewith, we have sequenced the near-complete coding region and part of the 3′UTR of the goat SREBF1 gene. In doing so, we have detected a missense c.353C>T polymorphism causing a proline to leucine substitution at position 118 (P118L). An association analysis with milk composition traits recorded in Murciano-Granadina goats only revealed a statistical tendency linking SREBF1 genotype and milk omega-3 fatty acid content. The lack of significant associations suggests that the P118L substitution does not involve a functional change., [FR]: Le facteur de transcription dénommé Sterol regulatory element binding transcription factor 1 (SREBF1) régule l'expression des gènes impliqués dans la biosynthèse des acides gras et du cholestérol. Dans cette étude, nous avons séquencé la quasi-totalité de la région codante et une partie du la région 3′UTR du gène SREBF1 de la chèvre. Ce travail, nous a permis d'identifier un polymorphisme non-synonyme c.353C> T causant la substitution d'une Proline en Leucine à la position 118. L’étude d'association avec la composition du lait enregistrée en chèvres Murciano-Granadina, a révélé seulement une tendance statistique reliant SREBF1 génotype et l'acide gras oméga-3 du lait. L'absence d'associations significatives suggère que la substitution P118L n'implique pas un changement fonctionnel.

Agouti signaling protein (ASIP) is one of the key players in the modulation of hair pigmentation in mammals. Binding to the melanocortin 1 receptor, ASIP induces the synthesis of phaeomelanin, associated with reddish brown, red, tan, and yellow coats. We have sequenced 2.8 kb of the goat ASIP gene in 48 individuals and identified two missense (Cys126Gly and Val128Gly) and two intronic polymorphisms. In silico analysis revealed that the Cys126Gly substitution may cause a structural change by disrupting a highly conserved disulfide bond. We studied its segregation in 12 Spanish and Italian goat breeds (N = 360) with different pigmentation patterns and found striking differences in the frequency of the putative loss-of-function Gly126 allele (Italian 0.43, Spanish Peninsular 0.08), but we did not observe a clear association with coat color. This suggests that the frequency of this putative loss-of-function allele has evolved under the influence of demographic rather than selection factors in goats from these two geographical areas.

In the lactating mammary gland, prolactin (PRL) stimulates the synthesis of lactose as well as fatty acid uptake, lipogenesis, and triacylglycerol synthesis. Associations between bovine PRL receptor (PRLR) genotype and fat yield have been reported, which illustrates the role of PRL in conveying lipids toward the udder as well as in stimulating their local synthesis during lactation. Conversely, and to the best of our knowledge, the effects of PRLR genotype on milk fatty acid content have not been studied so far in any mammalian species. In this study, we sequenced most of the coding region of the caprine PRLR gene in several individuals from the Malagueña and Murciano-Granadina breeds. This approach allowed us to identify 2 long and short mRNA isoforms, produced by alternative splicing, and 4 single-nucleotide polymorphisms (SNPs), namely, c.177T>C, c.1131G>A, c.1201G>A and c.1355C>T. Two of these SNPs are nonsynonymous and involve G401R (c.1201G>A) and T452I (c.1355C>T) amino acid substitutions in the cytoplasmic domain of PRLR, which plays a fundamental role in signal transduction. Performance of an association analysis with milk composition traits in a Murciano-Granadina goat population revealed highly suggestive effects on palmitoleic acid content, whereas suggestive effects were detected for other fatty acids, such as palmitic and linoleic. These results are consistent with the pleiotropic effects of PRL on mammary gland lipid metabolism and milk composition.

Malic enzyme 1 plays a fundamental role in lipid metabolism because it yields a significant amount of the NADPH necessary for fatty acid biosynthesis. In ruminants, however, its contribution to this biochemical process seems to be less relevant than in other livestock species. In this study, we have partially sequenced the goat ME1 gene with the aim of identifying polymorphic sites. Structural analysis of the goat ME1 amino acid partial sequence evidenced the existence of two dinucleotide-binding motifs, located at positions 158 to 163 (GLGDLG) and 301 to 306 (GAGEAA), and three amino acid residues (E245, D246 and D269) involved in the binding of Mn2+ and strongly conserved among mammalian species. Moreover, multiple sequence alignment allowed us to identify four single nucleotide polymorphisms at exons 5 (c.483C > T), 6 (c.667G > A), 9 (c.927C > T) and 11 (c.1200G > A). The effects of ME1 genotype on milk production and composition traits were investigated in a Murciano–Granadina goat population. None of the associations found reached statistical significance after applying Benjamini–Hochberg correction. At most, associations with uncorrected P-values below 0.01 were observed for C16:0, C18:1n-9t and total conjugated linoleic acids. These negative results reinforce the notion that ME1 plays an ancillary role in ruminant lipogenesis.

Hormone-sensitive lipase (LIPE) plays a fundamental role in the regulation of energy balance by releasing free fatty acids from adipose triacylglycerol stores. These fatty acids can be subsequently transferred to other body compartments to be oxidized or employed in other biochemical reactions. This enzymic function is particularly important in lactating animals because the synthesis of milk components involves the mobilization of lipid depots to satisfy the large energy demands of the mammary gland. In the current study, we partially sequenced the goat LIPE gene in several individuals. In doing so, we identified two synonymous polymorphisms at exons 2 (c.327C>A>T, triallelic polymorphism) and 3 (c.558C>T). Moreover, we found a mis-sense polymorphism at exon 6 (c.1162G>T) that involves an alanine to serine substitution at position 388. Analysis with Polyphen and Panther softwares revealed that this amino acid replacement is expected to be neutral. Performance of an association analysis with a variety of milk traits revealed that goat LIPE genotype has highly suggestive effects on milk yield (P=0·0032) as well as on C18:3 n-6g (P=0·0051), trans-10 cis-12 CLA (P=0·007) and C12:0 (P=0·0084) milk contents. These associations are concordant with the preference of LIPE to selectively mobilize medium-chain and unsaturated fatty acids.

Genetic variability of the caprine stearoyl-CoA desaturase 1 (SCD1) gene has been investigated by sequencing a 4.7-kb cDNA in 6 goats from the Murciano-Granadina and Malagueña breeds. Sequence alignment revealed the existence of one synonymous polymorphism at exon 5 (c.732C > T) and one nucleotide substitution (c.*3504G > A) at exon 6 that encodes the 3′ untranslated region (UTR). Moreover, the existence of a previously reported 3′UTR polymorphism involving a 3-bp indel (c.*1902_1904delTGT) was confirmed. Single nucleotide polymorphism and haplotype-based association analyses revealed suggestive associations between genetic variability of the SCD1 locus and lactose, stearic, polyunsaturated, and conjugated linoleic fatty acid contents. Associations with milk fatty acid composition might be explained by the global effects that SCD1 exerts on mammary gland lipid metabolism through the down-modulation of key transcription factors. Interestingly, the performance of an in silico analysis revealed that the c.*1902_1904delTGT polymorphism involves a considerable change in the secondary structure of the SCD1 mRNA. Gene reporter assays and quantitative PCR analysis would be needed to assess if this mutation has a causal effect on milk polyunsaturated and conjugated linoleic fatty acid levels by altering the amount of SCD1 transcripts in mammary epithelial cells.

The polymorphism of major histocompatibility complex (MHC) class I genes has been often involved in the resistance/susceptibility to a variety of infectious and parasitic diseases. In this work, the complete sequence of the coding region of a major histocompatibility complex (MHC) class I gene in goats (Cahi-N*01701, GenBank accession no. EF569216) is reported. The length of the corresponding open reading frame was 1,077 bp encoding a mature protein of 337 amino acids. Sequencing of additional clones allowed us to identify a second locus (Cahi-NC4*50301, GenBank accession no. EF569217) that, after performing a Bayesian phylogenetic analysis, happened to cluster with a bovine non-classical MHC class I gene. Nonclassical MHC class I molecules display low levels of polymorphism and fulfill an important immunoregulatory role in the placenta to inhibit maternal rejection. This initial description of the gene content of the goat MHC class I region will contribute to the characterization, in this ruminant species, of one of the most important genetic factors in the elicitation of innate and adaptive immune responses against pathogens.