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Fontanesi L.,DIPROVAL | Speroni C.,DIPROVAL | Buttazzoni L.,Associazione Nazionale Allevatori Suini ANAS | Scotti E.,DIPROVAL | And 3 more authors.
Meat Science | Year: 2010

The aim of this work was to identify and analyse DNA markers in two cathepsin genes, cathepsin L (CTSL) and cathepsin S (CTSS), and study their association with meat quality parameters (including cathepsin activity of post mortem muscle) and several production traits in Italian Large White pigs. In silico analysis of the porcine CTSL and CTSS genes identified two new single nucleotide polymorphisms (SNPs), one located in exon 5 (CTSL) and one positioned in the 5′-untranslated region (CTSS). Allele frequencies of these two SNPs were evaluated in 7 different pig breeds. In addition, radiation hybrid mapping localized the CTSS gene on porcine chromosome 4, in an important QTL region for meat and fat deposition. Association analysis with meat quality traits, carried out in 268 Italian Large White pigs, indicated association of the CTSL polymorphism with average daily gain (ADG), weight of lean cuts (LC) and back fat thickness (BFT) estimated breeding values (P < 0.05). Results for LC and BFT were also confirmed using a selective genotyping approach in other Italian Large White pigs (P < 0.01). In the 268 Italian Large White pigs, the CTSS polymorphism was associated with feed:gain ratio and ADG (P < 0.05). No association was observed between the analysed markers and meat quality parameters (pH1, pHu, lactate, glycogen, glycolytic potential and cathepsin activity, all measured on Musculus semimembranosus). Overall these results, together with those previously reported for other genes of this family, suggest that cathepsin genes play a role in defining economically important traits in pigs. © 2010 Elsevier Ltd. All rights reserved.


Davoli R.,University of Bologna | Braglia S.,University of Bologna | Valastro V.,University of Bologna | Annarratone C.,University of Bologna | And 7 more authors.
Meat Science | Year: 2012

The melanocortin-4 receptor (MC4R) gene codes for a G protein transmembrane receptor playing an important role in energy homeostasis control. In pig a single nucleotide polymorphism c.1426G>A has been identified and associated to average daily gain, feed intake and fatness traits but a lack of agreement on the effects of the gene on carcass traits in different breeds comes out from many studies. In the present study the c.1426G>A polymorphism is analysed in two Italian pig breeds, Large White and Duroc to study the association of the MC4R gene with some carcass traits. The results show that the c.1426G>A polymorphism affects daily gain, feed conversion ratio and ham weight in both breeds, lean cuts in the Italian Duroc and backfat thickness in the Italian Large White. The presence of MC4R mRNA transcript in different porcine tissues was analysed. © 2011 Elsevier Ltd.


Zambonelli P.,University of Bologna | Davoli R.,University of Bologna | Bigi M.,University of Bologna | Braglia S.,University of Bologna | And 4 more authors.
BMC Genetics | Year: 2013

Background: The pH is an important parameter influencing technological quality of pig meat, a trait affected by environmental and genetic factors. Several quantitative trait loci associated to meat pH are described on PigQTL database but only two genes influencing this parameter have been so far detected: Ryanodine receptor 1 and Protein kinase, AMP-activated, gamma 3 non-catalytic subunit. To search for genes influencing meat pH we analyzed genomic regions with quantitative effect on this trait in order to detect SNPs to use for an association study. Results: The expressed sequences mapping on porcine chromosomes 1, 2, 3 in regions associated to pork pH were searched in silico to find SNPs. 356 out of 617 detected SNPs were used to genotype Italian Large White pigs and to perform an association analysis with meat pH values recorded in semimembranosus muscle at about 1 hour (pH1) and 24 hours (pHu) post mortem. The results of the analysis showed that 5 markers mapping on chromosomes 1 or 3 were associated with pH1 and 10 markers mapping on chromosomes 1 or 2 were associated with pHu. After False Discovery Rate correction only one SNP mapping on chromosome 2 was confirmed to be associated to pHu. This polymorphism was located in the 3'UTR of two partly overlapping genes, Deoxyhypusine synthase (DHPS) and WD repeat domain 83 (WDR83). The overlapping of the 3'UTRs allows the co-regulation of mRNAs stability by a cis-natural antisense transcript method of regulation. DHPS catalyzes the first step in hypusine formation, a unique amino acid formed by the posttranslational modification of the protein eukaryotic translation initiation factor 5A in a specific lysine residue. WDR83 has an important role in the modulation of a cascade of genes involved in cellular hypoxia defense by intensifying the glycolytic pathway and, theoretically, the meat pH value. Conclusions: The involvement of the SNP detected in the DHPS/WDR83 genes on meat pH phenotypic variability and their functional role are suggestive of molecular and biological processes related to glycolysis increase during post-mortem phase. This finding, after validation, can be applied to identify new biomarkers to be used to improve pig meat quality. © 2013 Zambonelli et al.


Davoli R.,University of Bologna | Gandolfi G.,University of Bologna | Braglia S.,University of Bologna | Comella M.,University of Bologna | And 3 more authors.
Molecular Biology Reports | Year: 2011

PLIN2 (perilipin 2) is a cytosolic protein that promotes the formation and stabilization of the intracellular lipid droplets, organelles involved in the storage of lipid depots. Porcine PLIN2 gene represents a biological and positional candidate for fat deposition, a polygenic trait that affects carcass and meat quality. The aim of the present study was to screen PLIN2 gene for polymorphisms, to evaluate the association with carcass quality traits, and to investigate the gene expression in skeletal muscle. Six new single nucleotide polymorphisms (SNP) were detected by sequencing 32 samples from five pig breeds (Italian Large White, Italian Duroc, Italian Landrace, Belgian Landrace, Pietrain). Two SNP localized in introns, two in the 3′-untranslated region (UTR), and two missense SNP were found in exons. A 3′-UTR mutation (GU461317:g.98G>A), genotyped in 290 Italian Duroc pigs by High Resolution Melting, resulted significantly associated (P < 0.01) with average daily gain, feed conversion ratio, lean cuts and hams weight estimated breeding values. PLIN2 gene expression analysis in skeletal muscle of Italian Large White and Italian Duroc pigs divergent for backfat thickness and visible intermuscular fat showed a trend of higher expression level in pigs with higher intermuscular fat. These results suggest that PLIN2 can be a marker for carcass quality in pigs. Further investigation at both gene and protein level could elucidate its role on fat deposition. © 2010 Springer Science+Business Media B.V.


Fontanesi L.,University of Bologna | Scotti E.,University of Bologna | Buttazzoni L.,Associazione Nazionale Allevatori Suini ANAS | Dall'Olio S.,University of Bologna | And 4 more authors.
Molecular Biology Reports | Year: 2010

We recently showed that a polymorphism in the fat mass and obesity associated (FTO) gene (AM931150: g.276T > G) is associated with fat deposition traits in pigs. To confirm this result, we genotyped this polymorphism in an Italian Duroc population made up by 313 performance tested pigs with known estimated breeding values (EBVs) for average daily gain, back fat thickness (BFT), feed:gain ratio, lean cuts (LC), and visible intermuscular fat (VIF, a measure of intermuscular fat in the hams). In addition, we genotyped 148 commercial heavy pigs for which several fat deposition traits and lean meat percentage were measured. The results of the association analyses confirmed the effect of the FTO mutation on obesity-related traits (VIF, BFT and LC) in the Italian Duroc pigs (P < 0.01) and in the commercial pigs (intramuscular fat content of different muscles, P < 0.05 or P < 0.10; lean meat content, P < 0.05; BFT, P < 0.05; intermuscular fat content in the hams, P < 0.05). © 2009 Springer Science+Business Media B.V.


Fontanesi L.,University of Bologna | Scotti E.,University of Bologna | Buttazzoni L.,Associazione Nazionale Allevatori Suini ANAS | Dall'Olio S.,University of Bologna | And 2 more authors.
Molecular Biology Reports | Year: 2010

Cathepsin K (CTSK) was selected as a candidate gene for fat deposition in pigs because recently, in human and mouse, it was shown that this lysosomal proteinase is an obesity marker. A single nucleotide polymorphism (SNP) was identified in intron 4 of the porcine CTSK gene (g.15G>A; FM209043). Allele frequencies of this polymorphism were analysed in seven pig breeds. Radiation hybrid mapping confirmed the localization of CTSK to porcine chromosome 4, close to the FAT1 QTL region. Three populations of pigs (one Italian Large White and two Italian Duroc groups of pigs) were selected for association analysis. In the Italian Large White breed the g.15G>A SNP was not informative. Association analysis including all Italian Duroc pigs showed that the CTSK marker was associated with back fat thickness and lean cuts (P < 0.01), and average daily gain and feed:gain ratio (P < 0.05) estimated breeding values. © 2009 Springer Science+Business Media B.V.


Fontanesi L.,University of Bologna | Speroni C.,University of Bologna | Buttazzoni L.,Associazione Nazionale Allevatori Suini ANAS | Buttazzoni L.,Italian Agricultural Research Council | And 4 more authors.
Molecular Biology Reports | Year: 2012

We genotyped single nucleotide polymorphisms (SNPs) in 5 cathepsin or cystatin genes (cathepsin F, CTSF; cathepsin L, CTSL; cathepsin S, CTSS; cathepsin Z, CTSZ; cystatin B, CSTB) in two groups of Italian Duroc pigs: the first group (n. 100) was chosen using a selective genotyping approach with extreme estimated breeding value (EBV) for visible intermuscular fat (VIF); the second group (n. 218) was made of performance-tested Duroc pigs not selected by any criteria. CTSL marker showed a tendency towards association (P < 0.10) with VIF (first group) and back fat thickness (BFT) and average daily gain (ADG; second group). In the second group, the CTSL polymorphism was associated with weight of lean cuts (LC; P < 0.05). Additive effects for all mentioned traits in the second group was significant (P < 0.05). The results we obtained in the Italian Duroc pigs confirmed the results and the direction of the effects already reported for the Italian Large White breed. © 2011 Springer Science+Business Media B.V.


Fontanesi L.,University of Bologna | Bertolini F.,University of Bologna | Dall'Olio S.,University of Bologna | Buttazzoni L.,Italian Agricultural Research Council | And 2 more authors.
Animal Biotechnology | Year: 2012

In pigs, susceptibility to enterotoxigenic Escherichia coli (ETEC) K88 strains (locus F4bcR) is determined by a dominant allele, with the recessive allele determining resistance. The susceptible allele also appeared to be associated with higher growth rate even with discordant results. A single nucleotide polymorphism (SNP) in exon 7 of the mucin 4 (MUC4) gene (DQ848681:g.8227C>G), shown to be in close linkage disequilibrium with the F4bcR locus, has been used as marker to identify susceptible pigs, substituting invasive villous adhesion tests. We herein analyzed this SNP in Italian local breeds and applied a selective genotyping approach in Italian Large White, Italian Landrace, and Italian Duroc comparing allele frequency distribution in groups of pigs with extreme estimated breeding values (EBV) for average daily gain (ADG) and backfat thickness (BFT) to evaluate if this marker is associated with these traits. Allele G (associated with susceptibility to ETEC) was associated with higher ADG and BFT in Italian Large White (P=6.66E-04 and P=0.012, respectively) and higher ADG in Italian Landrace (P=7.23E-12). This polymorphism was poorly informative in Italian Duroc. Antagonistic associations of the MUC4g.8227C>G alleles on susceptibility to ETEC and growth performances evidence the complexity of applying marker assisted selection in pig breeding. © 2012 Taylor and Francis Group, LLC.


Fontanesi L.,University of Bologna | Speroni C.,University of Bologna | Buttazzoni L.,Associazione Nazionale Allevatori Suini ANAS | Scotti E.,University of Bologna | And 4 more authors.
Journal of Animal Science | Year: 2010

The objective of this study was to evaluate the effects of mutations in 2 genes [IGF2 and cathepsin D (CTSD)] that map on the telomeric end of the p arm of SSC2. In this region, an imprinted QTL affecting muscle mass and fat deposition was reported, and the IGF2 intron3-g.3072G>A substitution was identified as the causative mutation. In the same chromosome region, we assigned, by linkage mapping, the CTSD gene, a lysosomal proteinase, for which we previously identified an SNP in the 3′-untranslated region (AM933484, g.70G>A). We have already shown strong effects of this CTSD mutation on several production traits in Italian Large White pigs, suggesting a possible independent role of this marker in fatness and meat deposition in pigs. To evaluate this hypothesis, after having refined the map position of the CTSD gene by radiation hybrid mapping, we analyzed the IGF2 and the CTSD polymorphisms in 270 Italian Large White and 311 Italian Duroc pigs, for which EBV and random residuals from fixed models were calculated for several traits. Different association analyses were carried out to distinguish the effects of the 2 close markers. In the Italian Large White pigs, the results for IGF2 were highly significant for all traits when using either EBV or random residuals (e.g., using EBV: lean cuts, P = 2.2 × 10-18; ADG, P = 2.6 × 10-16; backfat thickness, P = 2.2 × 10-9; feed:gain ratio, P = 2.3 × 10-9; ham weig ht, P = 1.5 × 10-6). No effect was observed for meat quality traits. The IGF2 intron3-g.3072G>A mutation did not show any association in the Italian Duroc pigs, probably because of the small variability at this polymorphic site for this breed. However, a significant association was evident for the CTSD marker (P < 0.001) with EBV of all carcass and production traits in Italian Duroc pigs (lean content, ADG, back-fat thickness, feed:gain ratio) after excluding possible confounding effects of the IGF2 mutation. The effects of the CTSD g.70G>A mutation were also confirmed in a subset of Italian Large White animals carrying the homozygous genotype IGF2 intron3-g.3072GG, and by haplotype analysis between the markers of the 2 considered genes in the complete data set. Overall, these results indicate that the IGF2 intron3-g.3072G>A mutation is not the only polymorphism affecting fatness and muscle deposition on SSC2p. Therefore, the CTSD g.70G>A polymorphism could be used to increase selection efficiency in marker-assisted selection programs that already use the IGF2 mutation. However, for practical applications, because the CTSD gene should not be imprinted (we obtained this information from expression analysis in adult skeletal muscle), the different modes of inheritance of the 2 genes have to be considered. © 2010 American Society of Animal Science.


Fontanesi L.,University of Bologna | Colombo M.,University of Bologna | Tognazzi L.,University of Bologna | Scotti E.,University of Bologna | And 4 more authors.
Molecular Biology Reports | Year: 2011

TBC1D1 [TBC1 (tre-2/USP6, BUB2, cdc16) domain family, member 1] is a Rab-GTPase-activating related protein implicated in regulating the trafficking of glucose transporter 4 (GLUT4 or SLC2A4) storage vesicles to the cell surface in response to insulin and AMPK-activating stimuli in skeletal muscle. Mutations in the human and mouse TBC1D1 genes confer risk of obesity or leanness. We identified five single nucleotide polymorphisms (SNPs) in the porcine TBC1D1 gene. One of them (FN677935:g.219G>A) was genotyped either by high resolution melting and PCR-RFLP analyses to study allele frequencies in a few pig breeds and evaluate association with meat production and carcass traits in five groups of sib-tested pigs of Italian Large White and Italian Duroc breeds. The g.219G>A SNP was associated (P < 0.05) with ham weight, back fat thickness and lean cuts content in Italian Large White and with visible intermuscular fat in Italian Duroc pigs. © 2010 Springer Science+Business Media B.V.

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