Lean Pigs Research Articles

Energy Dense, Protein Restricted Diet Increases Adiposity and Perturbs Metabolism in Young, Genetically Lean Pigs

Animal models of obesity and metabolic dysregulation during growth (or childhood) are lacking. Our objective was to increase adiposity and induce metabolic syndrome in young, genetically lean pigs. Pre-pubertal female pigs, age 35 d, were fed a high-energy diet (HED; n = 12), containing 15% tallow, 35% refined sugars and 9.1–12.9% crude protein, or a control corn-based diet (n = 11) with 12.2–19.2% crude protein for 16 wk. Initially, HED pigs self-regulated energy intake similar to controls, but by wk 5, consumed more (P<0.001) energy per kg body weight. At wk 15, pigs were subjected to an oral glucose tolerance test (OGTT); blood glucose increased (P<0.05) in control pigs and returned to baseline levels within 60 min. HED pigs were hyperglycemic at time 0, and blood glucose did not return to baseline (P = 0.01), even 4 h post-challenge. During OGTT, glucose area under the curve (AUC) was higher and insulin AUC was lower in HED pigs compared to controls (P = 0.001). Chronic HED intake increased (P<0.05) subcutaneous, intramuscular, and perirenal fat deposition, and induced hyperglycemia, hypoinsulinemia, and low-density lipoprotein hypercholesterolemia. A subset of HED pigs (n = 7) was transitioned back to a control diet for an additional six weeks. These pigs were subjected to an additional OGTT at 22 wk. Glucose AUC and insulin AUC did not improve, supporting that dietary intervention was not sufficient to recover glucose tolerance or insulin production. These data suggest a HED may be used to increase adiposity and disrupt glucose homeostasis in young, growing pigs.

Sirt1 Inhibits Akt2-Mediated Porcine Adipogenesis Potentially by Direct Protein-Protein Interaction

Compared with the rodent, the domestic pig is a much better animal model for studying adipogenesis and obesity-related diseases. Currently, the role of Akt2 and Sirt1 in porcine adipogenesis remains elusive. In this study, we defined the effect of Akt2 and Sirt1 on porcine preadipocyte lipogenesis and the regulatory mechanism. First, we found that Akt2 was widely expressed in porcine various tissues and at high level in adipose tissue. Further analysis showed that the expression level of Akt2 was much higher in adipose tissue and adipocytes of the Bamei pig breed (a Chinese indigenous fatty pig) than in that of the Large White pig breed (a Lean type pig), whereas the level of Sirt1 expression was opposite. The expression levels of Sirt1 and Akt2 gradually increased during adipogenic differentiation. Adipogenesis was robustly inhibited in Akt2 deficient fat cells, whereas it was promoted in Sirt1 deficient cells using the lentiviral–mediated shRNA approach. Interestingly, adipogenesis returned to normal in Akt2 and Sirt1 dual–deficient cells, showing that the pro- and anti–adipogenic effects were balanced. Sirt1 inhibited transcriptional activity of Akt2 in a dose-dependent way. Interaction of endogenous Akt2 and Sirt1 was gradually enhanced before day 6 of differentiation, and then attenuated. Akt2 and Sirt1 also interacted with C/EBPα in adipocytes. Moreover, knockdown of Akt2 or/and Sirt1 affected pro–lipogenesis of insulin–stimulated by PI3K/Akt pathway. We further found that Sirt1 respectively interacted with PI3K and GSK3β which were key upstream and downstream components of PI3K/Akt pathway. Based on the above findings, we concluded that the crosstalk between C/EBPα and PI3K/Akt signaling pathways is implicated in Akt2 and Sirt1 regulation of adipogenesis.

Breed, sex and anatomical location-specific gene expression profiling of the porcine skeletal muscles

BackgroundSkeletal muscle is one of the most important economic traits in agricultural animals, especially in pigs. In the modern pig industry, lean type pigs have undergone strong artificial selection for muscle growth, which has led to remarkable phenotypic variations compared with fatty type pigs, making these different breeds an ideal model for comparative studies.ResultsHere, we present comprehensive gene expression profiling for the white (longissimus dorsi muscle) and the red (psoas major muscle) skeletal muscles among male and female fatty Rongchang, feral Tibetan and lean Landrace pigs, using a microarray approach. We identified differentially expressed genes that may be associated the phenotypic differences of porcine muscles among the breeds, between the sexes and the anatomical locations. We also used a clustering method to identify sets of functionally coexpressed genes that are linked to different muscle phenotypes. We showed that, compared with the white muscles, which primarily modulate metabolic processes, the red muscles show a tendency to be a risk factor for inflammation and immune-related disorders.ConclusionsThis analysis presents breed-, sex- and anatomical location-specific gene expression profiles and further identified genes that may be associated with the phenotypic differences in porcine muscles among breeds, between the sexes and the anatomical locations.

Vascular transcriptional alterations produced by juvenile obesity in Ossabaw swine

We adopted a transcriptome-wide microarray analysis approach to determine the extent to which vascular gene expression is altered as a result of juvenile obesity and identify obesity-responsive mRNAs. We examined transcriptional profiles in the left anterior descending coronary artery (LAD), perivascular fat adjacent to the LAD, and descending thoracic aorta between obese (n = 5) and lean (n = 6) juvenile Ossabaw pigs (age = 22 wk). Obesity was experimentally induced by feeding the animals a high-fat/high-fructose corn syrup/high-cholesterol diet for 16 wk. We found that expression of 189 vascular cell genes in the LAD and expression of 165 genes in the thoracic aorta were altered with juvenile obesity (false discovery rate ≤ 10%) with an overlap of only 28 genes between both arteries. Notably, a number of genes found to be markedly upregulated in the LAD of obese pigs are implicated in atherosclerosis, including ACP5, LYZ, CXCL14, APOE, PLA2G7, LGALS3, SPP1, ITGB2, CYBB, and P2RY12. Furthermore, pathway analysis revealed the induction of proinflammatory and pro-oxidant pathways with obesity primarily in the LAD. Gene expression in the LAD perivascular fat was minimally altered with juvenile obesity. Together, we provide new evidence that obesity produces artery-specific changes in pretranslational regulation with a clear upregulation of proatherogenic genes in the LAD. Our data may offer potential viable drug targets and mechanistic insights regarding the molecular precursors involved in the origins of overnutrition and obesity-associated vascular disease. In particular, our results suggest that the oxidized LDL/LOX-1/NF-κB signaling axis may be involved in the early initiation of a juvenile obesity-induced proatherogenic coronary artery phenotype.

Genetic parameters for androstenone and skatole as indicators of boar taint and their relationship to production and litter size traits in Danish Landrace1

Boar taint is an offensive odor that affects the smell and taste of cooked pork, resulting mainly from the accumulation of skatole and androstenone in the back fat of intact males. The aim of the study was to estimate genetic parameters for skatole and androstenone and their genetic relationship to production and litter size traits. Concentrations of skatole and androstenone in the back fat were available for approximately 6,000 and 1,000 Landrace boars, respectively. The concentrations were log-transformed to align phenotypic measures to a normal distribution. Heritability estimates for Log(skatole) and Log(androstenone) were 0.33 and 0.59, respectively. The genetic correlation between the 2 measures of boar taint was 0.37, suggesting that genetic selection against boar taint based on only 1 of the chemical compounds could be insufficient. The boar taint compounds had low and mostly favorable genetic correlations with the production traits. Most noticeable, a favorable genetic correlation of -0.20 between meat percentage and Log(skatole) was estimated and hence continued selection for lean pigs can also slowly reduce the level of boar taint if the desired carcass weight is kept constant. The relationship between litter size traits (measured on sows related to boars) and boar taint compounds was low and not significantly different from 0. In conclusion, skatole and androstenone can be reduced through selection without affecting important economical production and litter size traits. Therefore, animal breeding offers an effective and sustainable solution to surgical castration of male piglets.

Differential effects of reduced protein diets on fatty acid composition and gene expression in muscle and subcutaneous adipose tissue of Alentejana purebred and Large White × Landrace × Pietrain crossbred pigs

The present study assessed the effect of pig genotype (fatty v. lean) and dietary protein and lysine (Lys) levels (normal v. reduced) on intramuscular fat (IMF) content, subcutaneous adipose tissue (SAT) deposition, fatty acid composition and mRNA levels of genes controlling lipid metabolism. The experiment was conducted on sixty intact male pigs (thirty Alentejana purebred and thirty Large White × Landrace × Pietrain crossbred), from 60 to 93 kg of live weight. Animals were divided into three groups fed with the following diets: control diet equilibrated for Lys (17·5 % crude protein (CP) and 0·7 % Lys), reduced protein diet (RPD) equilibrated for Lys (13·2 % CP and 0·6 % Lys) and RPD not equilibrated for Lys (13·1 % CP and 0·4 % Lys). It was shown that the RPD increased fat deposition in the longissimus lumborum muscle in the lean but not in the fatty pig genotype. It is strongly suggested that the effect of RPD on the longissimus lumborum muscle of crossbred pigs is mediated via Lys restriction. The increase in IMF content under the RPD was accompanied by increased stearoyl-CoA desaturase (SCD) and PPARG mRNA levels. RPD did not alter backfat thickness, but increased the total fatty acid content in both lean and fatty pig genotype. The higher amount of SAT in fatty pigs, when compared with the lean ones, was associated with the higher expression levels of ACACA, CEBPA, FASN and SCD genes. Taken together, the data indicate that the mechanisms regulating fat deposition in pigs are genotype and tissue specific, and are associated with the expression regulation of the key lipogenic genes.

Effect of breed on the expression of Sirtuins (Sirt1-7) and antioxidant capacity in porcine brain

Sirtuins, NAD-dependent histone deacetylase (HDAC), are correlated to aging and antioxidant. The aim of this study was to determine breed differences of porcine Sirtuins expression and antioxidant capacity in brain between Jinhua pigs (a fatty breed of China) and Danish Landrace pigs (a leaner breed). Effect of age on Sirtuins’ expression was also investigated. At the age of 180 days, the mRNA levels of Sirt1, as well as Sirt2 and Sirt4, in Jinhua pigs were greater, but the mRNA levels of Sirt3, Sirt5, Sirt6, and Sirt7 of Jinhua pigs were lower compared with Danish Landrace pigs. Likewise, at the same BW of 64 kg, the mRNA levels of Sirtuins, except Sirt5 and Sirt7, in Jinhua pigs were greater than Danish Landrace pigs. Meanwhile, Jinhua pigs possessed higher antioxidants activity than Danish Landrace pigs either at the same age or at the same BW. Furthermore, mRNA levels of Sirtuins were decreased with age in brain of the two breeds from 30 to 120 days. The results indicated that Sirtuins expression in brain was different between fatty and lean pigs, and Sirtuins expression may be correlated to antioxidant capacity. In addition, age could down-regulate Siruins expression in porcine brain.

Fatty Acid and Transcriptome Profiling of Longissimus Dorsi Muscles between Pig Breeds Differing in Meat Quality

Fat and lean pig breeds show obvious differences in meat quality characteristics including the fatty acid composition of muscle. However, the molecular mechanism underlying these phenotypes differences remains unknown. This study compared meat quality traits between Lantang (a Chinese indigenous breed) and Landrace (a typical lean breed). The Lantang pigs showed higher L* values and intramuscular fat content, lower pH45min, pH24h and shear force in longissimus dorsi (LD) muscle than Landrace (P < 0.05). Fatty acid analysis demonstrated the lower monounsaturated fatty acids (MUFA) and higher polyunsaturated fatty acids (PUFA) percentage in Lantang LD than that in Landrace LD (P < 0.05). To further identify candidate genes for fatty acid composition, the transcriptome of LD muscle from the two breeds were measured by microarrays. There were 586 transcripts differentially expressed, of which 267 transcripts were highly expressed in Lantang pigs. After the validation by real-time quantitative PCR, 13 genes were determined as candidate genes for fatty acid composition of muscle, including Stearoyl-CoA desaturase (SCD). Then, a SCD over-expression plasmid was transfected into C2C12 cells to reveal the effect of SCD on the fatty acid composition in vitro. The results showed that SCD over-expression significantly increased PUFA proportion, while reduced that of saturated fatty acids (SFA) in C2C12 cells (P < 0.05). In summary, this study compared the differences of fatty acid composition and transcriptome in two breeds differing in meat quality, and further identified the novel role of SCD in the regulation of PUFA deposition.

Soft tissue calcification in the Ossabaw miniature pig: experimental and kinetic modeling studies

Calcium (Ca) deposition into vascular tissue was measured in Ossabaw miniature pigs with and without metabolic syndrome (MetS) using Ca tracer kinetics and coronary atherosclerosis measured with intravascular ultrasound. Pigs with MetS had higher Ca uptake into coronary arteries than lean pigs. Ca deposition into arteries is a common disease in humans. The Ossabaw pig develops MetS when fed an atherogenic diet. The aim of this study was to measure Ca deposition into arteries of lean vs. MetS pigs. Male pigs were fed for 5 months with chow diet (healthy, lean; n = 7) or atherogenic diet (n = 8) consisting of chow supplemented with 2 % cholesterol, 43 % kcal from fat, and 20 % kcal from fructose. Pigs were verified to have MetS by obesity, insulin resistance, impaired glucose tolerance, dyslipidemia, and hypertension. Two pigs received 50 nCi of (41)Ca i.v. and blood was drawn frequently for 24 h, and 2, 3, 6, 8, 10, 15, 20, and at sacrifice at 28 days after injection. Peripheral arteries were biopsied four times per pig over the 28th day and coronary artery sampled at sacrifice. Tissues were analyzed for (41)Ca:Ca. A compartmental model was used to estimate rates of Ca deposition into the arteries. The MetS swine had higher (41)Ca and atherosclerosis in coronary arteries than lean pigs. This pig model is a suitable model for studying vascular calcification in humans.

Orosomucoid expression profiles in liver, adipose tissues and serum of lean and obese domestic pigs, Göttingen minipigs and Ossabaw minipigs

The acute phase protein orosomucoid (ORM) has anti-inflammatory and immunomodulatory effects, and may play an important role in the maintenance of metabolic homeostasis in obesity-induced low-grade inflammation. Even though the pig is a widely used model for obesity related metabolic symptoms, the expression of ORM has not yet been characterized in such pig models. The objective of this study was to investigate the expression of ORM1 mRNA in liver, visceral adipose tissue, subcutaneous adipose tissue (SAT) from the abdomen or retroperitoneal abdominal adipose tissue (RPAT) and SAT from the neck, as well as the serum concentration of ORM protein in three porcine obesity models; the domestic pig, Göttingen minipigs and Ossabaw minipigs.No changes in ORM1 mRNA expression were observed in obese pigs compared to lean pigs in the four types of tissues. However, obese Ossabaw minipigs, but none of the other breeds, showed significantly elevated ORM serum concentrations compared to their lean counterparts. Studies in humans have shown that the expression of ORM was unchanged in adipose tissue depots in obese humans with an increased serum concentration of ORM. Thus in this respect, obese Ossabaw minipigs behave more similarly to obese humans than the other two pig breeds investigated.

Comparison of the longissimus muscle proteome between obese and lean pigs at 180 days

Production of high-quality meat is important to satisfy the consumer and make the pig industry competitive. Obese and lean breeds of pig show clear differences in adipogenic capacity and meat quality, but the underlying molecular mechanism remains unclear. We have compared protein expression of the longissimus muscle between Lantang (LT, obese) and Landrace (LR, lean) pigs at the age of 180days using two-dimensional fluorescence difference gel electrophoresis. Of the 1,400 protein spots detected per gel, 18 were differentially expressed between the two breeds. Using peptide mass fingerprint and tandem mass spectrometry, 17 protein spots were identified, corresponding to ten different proteins that could be divided into four groups: metabolism-related, structure-related, stress-related, and other (unclassified). Among the metabolism-related proteins, COX5A and ATP5B, which participate in oxidative phosphorylation, were highly expressed in LT, whereas ENO3, which is involved in glycolysis, was highly expressed in LR. These results may contribute valuable information to our understanding of the molecular mechanism responsible for differences between obese and lean pigs, such as growth rate and meat quality.

A field assessment of the effect of pre-slaughter conditions and genetic-stress susceptibility on blood welfare indicators in pigs

AbstractThe effect of pre-slaughter handling conditions and the RYR1 gene on blood Cortisol, lactate and creatine Phosphokinase (CPK) levels at exsanguination were assessed using 2,923 surveyed pigs from 106 deliveries to five Spanish abattoirs across two seasons. The relationship between blood parameters, carcase skin damage and pork quality traits was also assessed. The season influenced blood cortisol, lactate and CPK values. Females always showed higher concentrations of cortisol, lactate, and CPK than males. Pigs carrying the recessive allele of the RYR1 gene exhibited increased lactate and CPK concentrations but not cortisol. The cortisol concentration decreased in lean pigs that were slaughtered in winter after short lairage periods. The lactate concentration decreased with loading time and increased in summer with lairage time and carcase lean content. The CPK concentration increased with lairage time, carcase weight, and carcase lean content, and with the duration of winter transports. Each truck delivery only explained approximately 10% of the variance in blood parameters. Lairage time is the most influential pre-slaughter handling practice on the assessed welfare indicators. In addition, different optimal lairage times might be appropriate depending on season. Blood cortisol, lactate, and CPK concentrations increased concomitantly with skin damage score. Blood parameters were weakly correlated and they also showed low association with pork quality traits.

Effects of low protein diets on lean genotype pigs – growth, feed/ N-intake and fat indicators

The EU nitrate directive and the increasing cost of protein sources are leading farmers to reduce the nitrogen content in livestock feed. UK pig production often employs high protein ration to ensure high growth rate and low fat deposition. The aim was to compare the performances of pigs of a lean genotype fed with a conventional (C) or 2 low protein (LP) diets, lysine supplemented (LP1) or not (LP2). 64 animals on each diet were reared from 40 to 115 kg and fed ad-libitum. Liveweights (LW) and feed intake (FI) were recorded and after slaughter backfat (P2 site) thickness was measured and samples of longissimus muscle were analysed for total fatty acids. Pen-based data were analysed examining diet and batch as main factors. There were no significant diet effects on FI. Pigs on LP2 had a lower average daily weight gain (ADG) and higher feed conversion ratio (FCR) than C or LP1 from 60 kg onwards. Between diet strategies there were no significant differences in backfat thickness, but body fat deposition was higher in the LP2 group, followed by the LP1, and C the lowest. Results confirm that the LP1 strategy allows growth performance similar to the C diet but with 11% higher intramuscular fat (IMF). Pigs on LP2 diet show an increased body fat and IMF, although subcutaneous fat thickness was little affected. LP1 results indicate that reduced nitrogen intake (C vs. LP1 by 11 to 15%) can be achieved without compromising the growth performance, however feed conversion is significantly poorer (−6%) compared to the C diet probably due to amino acids (AA) deficiencies.

Genetic parameters for fatness-related traits in pigs: animal model for human obesity

Heritabilities and genetic correlations were calculated using a BLUP-animal model, taking age (when appropriate), batch and sex as fixed effects into account. An F2 population consisting of 450 pigs was created using a cross between Yorkshire and Duroc * Gottingen minipig. The Duroc and Yorkshire pigs are typically lean pigs, selected for decades for a low fat content, leanness and high growth rate. The Gottingen minipigs are bred principally for its small size and are known to be to obesity.The resulting F2 cross shows a wide range of phenotypic characteristics.

Response analysis of the Iberian pig growing from birth to 150 kg body weight to changes in protein and energy supply1

A total of 251 growing-finishing Iberian (IB) pigs, 32 of which were suckling piglets, were used in 5 separate sets of trials. The comparative slaughter procedure was used to determine nutrient and energy retention at several stages of growth from birth to 150 kg BW. A factorial arrangement was used within each set of trials, involving several concentrations of ideal protein in the diets as 1 factor and 2 or 3 levels of feed intake as the other. The main objective of these studies was to derive the optimal protein-to-energy ratio in the diet to allow for the expression of maximum protein deposition rates. The effect of feed restriction on growth performance, protein deposition, and fat deposition was also assessed. According to allometric equations, empty BW (EBW) was related to whole body components or total chemical constituents of empty body mass (P < 0.001). For pigs receiving solid feed, highly statistically significant multiple regression equations were constructed, which derived nutrient (g/kg) or energy (MJ/kg) composition as a function of EBW, dietary protein-to-energy ratio, and level of feeding (P < 0.001). In pigs offered adequate protein-to-energy diets, ADG at each stage of production was predicted as a function of the average BW and feeding level (P < 0.001). It was observed that the estimates of ME required for maintenance and net efficiency of utilization of ME for growth change were within rather narrow ranges throughout the growth stages studied. Preferred values (413 kJ/kg BW(0.75) × d(-1) and 0.593 for ME(m) and k(g), respectively) were obtained by regressing total energy retention (kJ/kg BW(0.75) × d(-1)) against ME intake (kJ/kg BW(0.75) × d(-1)). A multiple-regression approach revealed that in the IB pig, ME costs for protein deposition and fat deposition reach 60 and 62 kJ/g, which is considerably greater than in conventional or lean pig genotypes. In the IB pig, the maximum daily rate of protein deposition (PD(max), g) seemed to follow a linear-plateau shape with a breaking point at 32.5 kg BW, beyond which PD(max) remained at an average rate of 75 g × d(-1). The marginal efficiency of body protein deposition was estimated at each growth stage. In pigs fed on optimal or suboptimal protein-to-energy diets, the relationship between PD and ME intake declined, following a curvilinear pattern with increasing BW; thus, implying relative increases in lipid gain as BW increased.

Differences in lipid deposition and adipose membrane biophysical properties from lean and obese pigs under dietary protein restriction

Obesity consists in fat accumulation leading to increase in adipose cells number and size. Adipocyte membrane biophysical properties are critical to maintain cellular viability in metabolically healthy obesity. This study investigated the effect of the genetic background and dietary protein restriction on fat tissue lipid composition, adipocyte membrane fluidity and water permeability using the pig as experimental model.Twenty-four male pigs from distinct genotypes, lean and obese, were fed on normal and reduced protein diets within a 2×2 factorial arrangement (two genotypes and two diets). Backfat thickness was twofold higher in obese than in lean pigs but unrelated to dietary protein level. In contrast, total fatty acids in the subcutaneous adipose tissue were dependent on both breed and diet, with increased lipid content promoted by the fatty genotype and by the restriction of dietary protein. Adipose membranes isolated from obese pig’s subcutaneous fat tissue showed higher permeability to water, in line with an increased fluidity. Moreover, the reduced content of dietary protein influenced positively the fluidity of adipose membranes. Neither genotype nor diet affected total cholesterol concentration in the adipose membranes. Membrane-saturated fatty acids’ content was influenced by genotype, while membrane-polyunsaturated fatty acids, particularly from the n-6 family, was influenced by diet. The ratio of oleic (18:1c9)/linoleic (18:2n-6) acids was positively correlated with membrane fluidity. All together, these findings reinforce the genetic background as a determinant player on adipose membrane biophysical properties and point to the dietary protein level as an important factor for subcutaneous lipid deposition as well as for regulation of membrane function, factors that may have impact on human obesity and metabolic syndrome.

Impact of obesity on bone integrity and mesenchymal stem cell activity in Ossabaw swine

Ossabaw swine are an exciting model to study the effects of diet induced obesity. We examined the affects of an obesigenic diet on bone integrity and on mesenchymal stem cells (MSC) activity, which underpins bone integrity. Pigs fed an obesigenic diet had greater (P &lt; 0.05) body fat % and insulin resistance. Obese pigs had larger and heavier bones (P &lt; 0.05), however the percentage of mineral in the bone did not differ between obese and lean pigs. In order to evaluate alterations in MSC activity, MSC from obese and lean pigs were cultured under adipocytic and osteogenic conditions. Under osteogenic conditions, the expression of genes indicative of osteoblastic differentiation (RunX2 and osteocalcin) were greater (P&lt;0.1) in MSC isolated from obese pigs. Under adipogenic conditions, increased gene expression of markers of adipocytic differentiation (PPARG, AP2, and LPL), as well as greater lipid accumulation (Oil Red O staining), were seen in MSC isolated from the lean pigs. While the increased osteogenic activity of MSC from obese pigs supports the bone data, the reduced adipogenic activity of these cells seems counterintuitive. However, the reduced in vitro adipocytic differentiation of MSC from obese pigs may be a result of the insulin resistance seen in these pigs. These results suggest that obesity and insulin resistance may have long‐term effects on bone development and MSC activity.

Soft tissue calcification in the Ossabaw miniature pig: experimental and kinetic modeling studies

Early stage deposition of calcium into soft tissues is a pre‐cursor to several chronic diseases and an appropriate large animal model is needed. The Ossabaw pig develops metabolic syndrome when fed excess calorie atherogenic diet and progresses to type 2 diabetes and atherosclerosis. Male pigs were fed for 5 months with chow diet (health, lean; n=7) or atherogenic diet (n=8) consisting of chow supplemented with 2% cholesterol, 43% kcal from fat, and 20% kcal from fructose. Pigs were verified to have metabolic syndrome (MetS) by obesity, insulin resistance, impaired glucose tolerance, dyslipidemia, and hypertension. All pigs received intravenous injection of 50nCi/kg of 41Ca and allowed to recover for 1 month, then sacrificed. Blood was drawn at time 0, nearly hourly for the first 24 h, and 1, 2, 3, 6, 8, 10, 15, 20, and 28 days after injection. Peripheral artery segments were harvested from each pig at 4 time points over the 28d recovery. At sacrifice the MetS swine had detectable 41Ca in coronary arteries, but the lean controls had extremely low levels of 41Ca. Intravascular ultrasound showed calcification in MetS, but not lean pigs. A compartmental model of calcium metabolism was developed and deposition into arteries estimated. Through tissue acquisition and kinetics, this pig provides a humanoid model for studying vascular calcification.

Solexa Sequencing Identification of Conserved and Novel microRNAs in Backfat of Large White and Chinese Meishan Pigs

The domestic pig (Sus scrofa), an important species in animal production industry, is a right model for studying adipogenesis and fat deposition. In order to expand the repertoire of porcine miRNAs and further explore potential regulatory miRNAs which have influence on adipogenesis, high-throughput Solexa sequencing approach was adopted to identify miRNAs in backfat of Large White (lean type pig) and Meishan pigs (Chinese indigenous fatty pig). We identified 215 unique miRNAs comprising 75 known pre-miRNAs, of which 49 miRNA*s were first identified in our study, 73 miRNAs were overlapped in both libraries, and 140 were novelly predicted miRNAs, and 215 unique miRNAs were collectively corresponding to 235 independent genomic loci. Furthermore, we analyzed the sequence variations, seed edits and phylogenetic development of the miRNAs. 17 miRNAs were widely conserved from vertebrates to invertebrates, suggesting that these miRNAs may serve as potential evolutional biomarkers. 9 conserved miRNAs with significantly differential expressions were determined. The expression of miR-215, miR-135, miR-224 and miR-146b was higher in Large White pigs, opposite to the patterns shown by miR-1a, miR-133a, miR-122, miR-204 and miR-183. Almost all novel miRNAs could be considered pig-specific except ssc-miR-1343, miR-2320, miR-2326, miR-2411 and miR-2483 which had homologs in Bos taurus, among which ssc-miR-1343, miR-2320, miR-2411 and miR-2483 were validated in backfat tissue by stem-loop qPCR. Our results displayed a high level of concordance between the qPCR and Solexa sequencing method in 9 of 10 miRNAs comparisons except for miR-1a. Moreover, we found 2 miRNAs, miR-135 and miR-183, may exert impacts on porcine backfat development through WNT signaling pathway. In conclusion, our research develops porcine miRNAs and should be beneficial to study the adipogenesis and fat deposition of different pig breeds based on miRNAs.

Meat quality of the longissimus lumborum muscle of Casertana and Large White pigs: Metabolomics and proteomics intertwined

Longissimus lumborum muscles from high fat-deposing Casertana and lean meat Large White pigs were assayed for meat quality parameters, including early and ultimate post mortem pH, water holding capacity and Minolta L*a*b*values. These parameters were correlated to results from differential proteomic and targeted metabolomic analyses. Higher levels of glycolytic enzymes and lactate accumulation were related to slow pH drop in Casertana pigs, albeit not to rapid pH lowering in LW counterparts. On the other hand, the individuation of pyruvate kinaseM1 and tropomyosin levels in LW were related to water holding capacity and Minolta values at 24h after slaughter. Bioinformatic analyses strengthened the correlation between over-expression of structural proteins in LW and more accentuated growth aptitude in this breed. Conversely, enzymes taking part into branching glycolytic reactions, such as glycerol 3-phosphate and creatine kinase M, were related to accentuated lipogenesis and slower albeit prolonged glycolytic rate in Casertana, respectively. Breed-specific differences at the protein level were not only related to growth performances and fat accumulation tendency in vivo, but they also affected post mortem performances through a direct influence on the forcedly anaerobic behavior of pig muscles after slaughter.