Effect of Pig Domestication on Skeletal Muscle Development, Microstructure, and Genetic Mechanism Involved in Myofibre Type Formation

The wild boar and modern highly selected pigs are phenotypically distant European pig breeds reared in contrasting conditions and present ideal model to better understand the mechanisms behind meat quality deterioration related to domestication and selection pressure, which provoked substantial modifications in the ontogenic development as well as contractile and metabolic properties of skeletal muscles. The skeletal muscle of domestic pigs are less mature at birth and contains a lower number of myofibres compared to wild boars; however, expansive myofibre hypertrophy, protein accretion as well as additional myofibre formation are accelerated in the early postnatal period in some muscles in domestic pigs. A comparative view of the cellular and subcellular mechanisms underlying the skeletal myofibre development could help to design a breeding program that would improve the balance between the growth performance, muscularity and meat quality. This chapter therefore outlines the influence of domestication on myofibre formation and differentiation during growth and provides a comparative view on the developmental expression pattern of the MyHC isoforms, the activity of different metabolic enzymes, and the expression of selected genes responsible for the metabolic diversity of the myofibres. Additionally, there is a special emphasis on the type, composition, and histomorphological traits of myofibres

Effect of Immunocastration on Performance of Slovenian Pig Fatteners

The influence of the immunocastration (immunisation against GnRH) on pig performance (growth, carcass and meat quality traits) was investigated in two parallel experiments (on two farms) with two crossbreeds – G1 (50% Duroc) and G2 (50% Pietrain). Within the crossbreed, the pigs were assigned to three experimental groups; entire males (EM, n=49), immunocastrates (IC, n=45) and surgical castrates (SC, n=45). Those assigned to IC group were vaccinated at the age of 12 and 19 weeks. Pigs were individually housed, their feed intake (ad libitum) and weight (at 12, 19 and 24 weeks) were recorded. At the age of 24 weeks, the pigs were slaughtered and their carcass and meat quality traits were assessed. We hypothesized that treatment response could have been different in two crossbreeds. However the interaction was insignificant, thus the treatment effect is presented on pooled results for both crossbreeds. Until the revaccination, IC were similar to EM pigs, thereafter they exhibited an increase in feed intake and growth rate. Overall, they presented an advantage in growth rate and feed efficiency as compared to SC. They also exhibited better carcass properties as SC without any major effect on meat quality. The present study provides the initial information on the immunocastration effect in Slovenian herds that should further be supported by testing it in usual rearing conditions and group housing

Salivary Gland Adaptation to Dietary Inclusion of Hydrolysable Tannins in Boars

The ingestion of hydrolysable tannins as a potential nutrient to reduce boar odor in entire males results in the significant enlargement of parotid glands (parotidomegaly). The objective of this study was to characterize the effects of different levels of hydrolysable tannins in the diet of fattening boars (n = 24) on salivary gland morphology and proline-rich protein (PRP) expression at the histological level. Four treatment groups of pigs (n = 6 per group) were fed either a control (T0) or experimental diet, where the T0 diet was supplemented with 1% (T1), 2% (T2), or 3% (T3) of the hydrolysable tannin-rich extract Farmatan®. After slaughter, the parotid and mandibular glands of the experimental pigs were harvested and dissected for staining using Goldner’s Trichrome method, and immunohistochemical studies with antibodies against PRPs. Morphometric analysis was performed on microtome sections of both salivary glands, to measure the acinar area, the lobular area, the area of the secretory ductal cells, and the sizes of glandular cells and their nuclei. Histological assessment revealed that significant parotidomegaly was only present in the T3 group, based on the presence of larger glandular lobules, acinar areas, and their higher nucleus to cytoplasm ratio. The immunohistochemical method, supported by color intensity measurements, indicated significant increases in basic PRPs (PRB2) in the T3 and acidic PRPs (PRH1/2) in the T1 groups. Tannin supplementation did not affect the histo-morphological properties of the mandibular gland. This study confirms that pigs can adapt to a tannin-rich diet by making structural changes in their parotid salivary gland, indicating its higher functional activity

Obesity resistant mechanisms in the Lean polygenic mouse model as indicated by liver transcriptome and expression of selected genes in skeletal muscle

<p>Abstract</p> <p>Background</p> <p>Divergently selected Lean and Fat mouse lines represent unique models for a polygenic form of resistance and susceptibility to obesity development. Previous research on these lines focused mainly on obesity-susceptible factors in the Fat line. This study aimed to examine the molecular basis of obesity-resistant mechanisms in the Lean line by analyzing various fat depots and organs, the liver transcriptome of selected metabolic pathways, plasma and lipid homeostasis and expression of selected skeletal muscle genes.</p> <p>Results</p> <p>Expression profiling using our custom Steroltalk v2 microarray demonstrated that Lean mice exhibit a higher hepatic expression of cholesterol biosynthesis genes compared to the Fat line, although this was not reflected in elevation of total plasma or liver cholesterol. However, FPLC analysis showed that protective HDL cholesterol was elevated in Lean mice. A significant difference between the strains was also found in bile acid metabolism. Lean mice had a higher expression of <it>Cyp8b1</it>, a regulatory enzyme of bile acid synthesis, and the <it>Abcb11 </it>bile acid transporter gene responsible for export of acids to the bile. Additionally, a higher content of blood circulating bile acids was observed in Lean mice. Elevated HDL and upregulation of some bile acids synthesis and transport genes suggests enhanced reverse cholesterol transport in the Lean line - the flux of cholesterol out of the body is higher which is compensated by upregulation of endogenous cholesterol biosynthesis. Increased skeletal muscle <it>Il6 </it>and <it>Dio2 </it>mRNA levels as well as increased activity of muscle succinic acid dehydrogenase (SDH) in the Lean mice demonstrates for the first time that changes in muscle energy metabolism play important role in the Lean line phenotype determination and corroborate our previous findings of increased physical activity and thermogenesis in this line. Finally, differential expression of <it>Abcb11 </it>and <it>Dio2 </it>identifies novel strong positional candidate genes as they map within the quantitative trait loci (QTL) regions detected previously in crosses between the Lean and Fat mice.</p> <p>Conclusion</p> <p>We identified novel candidate molecular targets and metabolic changes which can at least in part explain resistance to obesity development in the Lean line. The major difference between the Lean and Fat mice was in increased liver cholesterol biosynthesis gene mRNA expression, bile acid metabolism and changes in selected muscle genes' expression in the Lean line. The liver <it>Abcb11 </it>and muscle <it>Dio2 </it>were identified as novel positional candidate genes to explain part of the phenotypic difference between the Lean and Fat lines.</p

Bone tissue morphology of rat offspring lactationally exposed to polychlorinated biphenyl 169 and 155

Polychlorinated biphenyls (PCBs) are ubiquitous, persistent, organic pollutants also considered endocrine-disrupting chemicals. Our study examined the effects of lactational exposure to nondioxin-like PCB-155 and/or dioxin-like PCB-169 on longitudinal femur growth at the distal epiphyseal growth plate (EGP) in young rats at three different ages [postnatal days (PNDs) 9, 22, and 42]. After delivery, lactating rats were divided into four groups (PCB-169, PCB-155, PCB-155 + 169, and control) and administered PCBs intraperitoneally. The femurs of offspring were used to estimate growth rate (µm/day), and histomorphometric analysis on the distal femur included the thickness of the EGP and zones of proliferation and hypertrophy with calcification. Stereometry was used to determine trabecular bone volume density. In the PCB-169 and PCB-155 + 169 groups, PCB-169 affected longitudinal bone growth in the early postnatal period by interfering with chondrocytes in the EGP zone of proliferation and, to a lesser extent, the zone of hypertrophy. Morphometric alterations in EGP structure diminished until puberty. A slow growth rate persisted in the PCB-169 group until PND 42, while in the PCB-155 group, a fast growth rate between PNDs 9 to 22 was significantly reduced between PNDs 22 to 42. Sterometric assessment showed decreased trabecular bone volume in the PCB-155 + 169 group compared with that in the control on PND 9 and increased in the PCB-169 group compared with that in the PCB-155 group on PND 42. To summarize, studied PCB congeners exerted congener- and age-dependent effects on femur growth rate and its histomorphometric characteristics

Age-Dependent Expression of MyHC Isoforms and Lipid Metabolism-Related Genes in the Longissimus Dorsi Muscle of Wild and Domestic Pigs

This study aimed to compare age-dependent changes in the relative expression of genes encoding myosin heavy chain (MyHC) isoforms and selected lipid metabolism-related genes in the longissimus dorsi muscle of wild pigs (WPs) and domestic pigs (DPs). Muscles sampled from postnatal day one as well as three-week-old and two-year-old animals were used in quantitative polymerase chain reaction (qPCR) assays, histological evaluations of succinate dehydrogenase (SDH) activity, and intra-myofiber lipid (IMFL) assessment. Expression of the MyHC isoforms displayed the most extensive age- and breed-dependent changes within the first three postnatal weeks. The MyHCembry level decreased significantly faster in the WPs than in the DPs. The relative MyHC-I and -IIa expression was significantly higher in the WPs, and MyHC-IIb was substantially higher in the DPs. The differences in MyHC expression corroborated the number of SDH-positive myofibers and IMFLs. Expression of the peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PGC-1α), peroxisome proliferator-activated receptor gamma (PPARγ) and lipoprotein lipase (LPL) genes displayed only age-related variations. In summary, the evidence is provided for accelerated postnatal myofiber transformation directed towards oxidative myofibers in WPs. The SDH activity/staining intensity largely reflected the expression of MyHCs, and not genes involved in lipid uptake and utilization

Effect of immunocastration on performance of Slovenian pig fatteners

The influence of the immunocastration (immunisation against GnRH) on pig performance (growth, carcass and meat quality traits) was investigated in two parallel experiments (on two farms) with two crossbreeds – G1 (50% Duroc) and G2 (50% Pietrain). Within the crossbreed, the pigs were assigned to three experimental groupsentire males (EM, n=49), immunocastrates (IC, n=45) and surgical castrates (SC, n=45). Those assigned to IC group were vaccinated at the age of 12 and 19 weeks. Pigs were individually housed, their feed intake (ad libitum) and weight (at 12, 19 and 24 weeks) were recorded. At the age of 24 weeks, the pigs were slaughtered and their carcass and meat quality traits were assessed. We hypothesized that treatment response could have been different in two crossbreeds. However the interaction was insignificant, thus the treatment effect is presented on pooled results for both crossbreeds. Until the revaccination, IC were similar to EM pigs, thereafter they exhibited an increase in feed intake and growth rate. Overall, they presented an advantage in growth rate and feed efficiency as compared to SC. They also exhibited better carcass properties as SC without any major effect on meat quality. The present study provides the initial information on the immunocastration effect in Slovenian herds that should further be supported by testing it in usual rearing conditions and group housing

Lactational exposure to dioxin-like polychlorinated biphenyl 169 and nondioxin-like polychlorinated biphenyl 155

Exposure to polychlorinated biphenyls (PCBs), which are persistent lipophilic environmental pollutants, has a variety of adverse effects on wildlife and human health, including bone mineralization, growth and mechanical strength. The present study evaluated the effects of lactational exposure to nondioxin-like PCB-155 and dioxin-like PCB-169, individually and in combination, on pubertal rat femur development and its biomechanics. After offspring delivery, Wistar rat mothers were divided into four groups, i.e., PCB-169, PCB-155, PCB-155+169 and control, and were administered PCBs intraperitoneally. Data on bone geometry, biomechanics and mineral composition were obtained by analysis of femurs from 42-day-old offspring by microCT scanning, three-point bending test and inductively coupled plasma mass spectrometry. Decreased somatic mass and femur size, i.e., mass, periosteal circumference and cross sectional area, were observed in the PCB-169 and PCB-155 groups. Additionally, lactational exposure to planar PCB-169 resulted in harder and more brittle bones containing higher amounts of minerals. Combined exposure to structurally and functionally different PCBs demonstrated only mild alterations in bone width and mineralization. To conclude, our results demonstrated that alterations, observed on postnatal day 42, were primarily induced by PCB-169, while toxicity from both of the individual congeners may have been reduced in the combined group

Adipose tissue gene expression of entire male, immunocastrated and surgically castrated pigs

Differences in adipose tissue deposition and properties between pig male sex categories, i.e., entire males (EM), immunocastrates (IC) and surgical castrates (SC) are relatively well-characterized, whereas the underlying molecular mechanisms are still not fully understood. To gain knowledge about the genetic regulation of the differences in adipose tissue deposition, two different approaches were used: RNA-sequencing and candidate gene expression by quantitative PCR. A total of 83 differentially expressed genes were identified between EM and IC, 15 between IC and SC and 48 between EM and SC by RNA-sequencing of the subcutaneous adipose tissue. Comparing EM with IC or SC, upregulated genes related to extracellular matrix dynamics and adipogenesis, and downregulated genes involved in the control of lipid and carbohydrate metabolism were detected. Differential gene expression generally indicated high similarity between IC and SC as opposed to EM, except for several heat shock protein genes that were upregulated in EM and IC compared with SC. The candidate gene expression approach showed that genes involved in lipogenesis were downregulated in EM compared with IC pigs, further confirming RNA-sequencing results