1Department of Animal Husbandry and Veterinary Medicine, Jiangsu Polytechnic College of Agriculture and Forestry, Jurong 212400, P. R. China 2Co-Innovation Center for Sustainable Forestry in Southern China, College of Forestry, Nanjing Forestry University, Nanjing 210037, P. R. China 3Key Laboratory of Animal Physiology & Biochemistry, Ministry of Agriculture, Nanjing Agricultural University, Nanjing 210095, P. R. China

Currently, the effects of stress on the body metabolism are attracting more and more attentions. To better understand the probable stress-induced metabolic changes, the changes in plasma indices and hepatic proteome were investigated in pigs fitted with jugular venous catheters administered adrenocorticotropic hormone (ACTH), intramuscularly twice daily for 7 days. In particular, plasma hormones, blood cells and biochemical indicators were analyzed. In addition, the hepatic proteome was analyzed by two-dimensional electrophoresis and matrix-assisted laser desorption ionization time of flight mass spectrometry. The results revealed that after the initial ACTH injection, plasma ACTH, cortisol, triiodothyronine and tumor necrosis factor α increased (P ˂ 0.05). Additionally, lactic dehydrogenase activity increased (P < 0.05), and both leukocyte and lymphocyte count decreased (P < 0.05). In comparison, after the ACTH injection for 7 days, hepatic proteomics analysis identified 27 significantly changed proteins and most of these proteins (49%) are involved in metabolism, but plasma indices showed no obvious change. In conclusion, compared with the obvious changes in plasma indices after the initial ACTH injection, the multiple ACTH injections did not change plasma indices, yet the hepatic proteome profile was significantly altered. The results provide useful information for a better understanding of the effects of stress on metabolism.

The objective of present study was to investigate changes in salivary components during restraint to identify potential markers of stress. Pigs were subjected to a nasal snare stress (Experiment 1) or an immobilization stress (Experiment 2) by being enclosed in a steel cage. Saliva was collected before, during and after the stress, respectively. Salivary cortisol, serum amyloid A (SAA), haptoglobin (HP), chromogranin A (CgA), amylase, K+, Ca2+ and lactoferrin content were detected. The results showed that in Experiment 1, HP and CgA content increased significantly at 10min during the restraint (P<0.05, P<0.05), in agreement with the significantly increased cortisol and SAA levels (P<0.01, P<0.05), while amylase, K+ and lactoferrin concentrations did not significantly change. In Experiment 2, salivary HP and CgA concentrations also changed significantly during the restraint (P<0.01, P<0.01), yet cortisol, SAA, amylase, K+ and lactoferrin levels did not show obvious change. The results confirmed that salivary HP and CgA content may be useful candidate biomarkers to monitor the physical state in pigs during stress.

The identification of useful biological indicators to monitor the body response before the presentation of clinical diseases has practical value in livestock production. To identify potential biomarkers in pigs at the early stage during inflammation, 12 pigs were intramuscularly injected with 2mL of Lipopolysaccharide (LPS, 15μg/kg BW) or saline. Serum protein expression profiles were detected with two-dimensional electrophoresis (2-DE) and matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) techniques. Serum biochemical indicators and acute phase protein (APP) concentrations were analyzed with an automatic biochemical analyzer and ELISA, respectively. Of the serum biochemical indicators, creatinine concentration significantly increased 6h post infection, whereas albumin showed a decreased tendency. The 2-DE and MALDI-TOF mass spectrometry technique detected 17 protein spots representing 10 proteins: α-1-antichymotrypsin, albumin, bovine lactoferrin, serotransferrin, serpin A3-6, immunoglobulin light chain (κ chain, mu chain), complement C3 precursor, zinc-α-2-glycoprotein (ZAG), and ceruloplasmin. Two proteins were selected to confirm the mass spectrometry results, and resulting differences accorded with the proteomics results. Of the four typical acute phase protein (APPs) measured, the C-reactive protein (CRP) and haptoglobin (HP) concentrations increased significantly, while no differences were observed in the pig-major acute phase protein (Pig-MAP) and transthyretin (TTR) contents. The results showed that serum creatinine, CRP, HP, and ten other proteins content changed significantly after LPS injection. Of these proteins, ZAG was first reported in pigs during inflammation. These proteins show great promise as biomarkers to monitor the health status and welfare of pigs during the early stage of LPS-induced inflammation.

BACKGROUND:In the practical commercial pig farms, inflammation is a perennial problem, yet most of studies on inflammation are focused on immune response. Actually, inflammation can induce body metabolism disorder which will finally influence animals' growth. In this study, we investigated the effect of acute inflammation on the triglyceride (TG) metabolism in the liver of growing pigs and the possible underlying mechanisms.METHODS:Twelve male growing pigs were randomly divided into two groups, a control group (received saline) and a LPS group (intramuscular injected with 15 μg/kg LPS). Six hours after LPS injection, the pigs were euthanized and sampled. Biochemical indexes, inflammation factors, lipid metabolism related parameters and mitochondrial function were evaluated. The relationship between glucocorticoid receptor (GR) and the key enzymes of de novo lipogenesis were also investigated by chromatin immunoprecipitation assay (ChIP).RESULTS:LPS induced a serious inflammation in the liver of growing pigs proved by liver morphologic changes, the up-regulated plasma cortisol, tumor necrosis factor-α (TNF-α) content and gene expression of inflammation related genes in liver. For de novo lipogenesis, LPS significantly decreased the gene expression of fatty acid synthase (FAS), Acetyl-CoA carboxylase-1 (ACC-1) and Stearoyl-CoA desaturase-1 (SCD-1), and the protein expression of ACC-1 and SCD-1. For lipolysis, only the gene expression of adipose triglyceride lipase (ATGL) was decreased. LPS did nothing to the gene expression of hormone-sensitive lipase (HSL) and the lipolytic enzymes activities. For β-oxidation, LPS significantly increased the protein expression of CPT-1α, but the gene expression of mitochondrial DNA-encoded genes and the activities of mitochondrial complex IV and V demonstrated no obviously changes. Furthermore, ChIP results showed that LPS significantly decreased the level of GR binding to ACC-1 promoter.CONCLUSION:LPS infection has a profound impact on hepatic TG metabolism. This impact is mainly demonstrated by the significantly deceased de novo lipogenesis, and GR may involve in its regulation.

The diagnostic use of saliva for disease detection and health monitoring is increasing in recent years. To investigate whether the components of saliva change with lipopolysaccharide (LPS) challenge and clarify whether these changes directly relate to the submandibular gland (SMG), 12 pigs were employed in the present study. At 6 h after LPS intramuscular injection, samples including saliva, plasma and SMG were taken. The amylase, cortisol, Immunoglobulin A (IgA), tumor necrosis factor alpha (TNF-α) and interleukin 1 beta (IL-1β) contents were analyzed by colorimetry, radioimmunoassay or ELISA, respectively. Chromogranin A (CgA) and lactoferrin protein concentrations were detected by western blot. Adenosine triphosphate (ATP) was measured by high-pressure liquid chromatography (HPLC) and the related gene expression was analyzed by reverse transcription polymerase chain reaction (RT-PCR) in SMG. The results showed that LPS treatment increased amylase activity and cortisol concentrations, whereas IL-1β, CgA and lactoferrin concentrations showed a decrease in saliva. Toll-like receptor 2 (TLR2), IL-1β and TNF-α gene expression in SMG increased. Decreased ATP concentration in SMG may be the reason for the opposite changes of IL-1β, lactoferrin and CgA content in the saliva and the SMG. The present study for the first time comprehensively demonstrates that the altered parameters in saliva can indicate the body inflammation in pigs, and they may become useful, noninvasive biomarkers for the evaluation of body state.

比较了老淮猪和杜长大三元杂交猪在生长和育肥两个不同年龄阶段血液中血细胞、常规生化指标、皮质醇激素、代谢酶类、急性期蛋白等指标的差异。双因素方差分析结果显示,两品种间红细胞数、红细胞压积及血红蛋白含量差异显著;红细胞数和血小板数目在不同年龄阶段差异显著。老淮猪血清高密度脂蛋白胆固醇显著高于三元杂交猪,但低密度脂蛋白的含量显著低于三元杂交猪。血清中常见的8种代谢酶类,均没有出现品种间差异。不同年龄阶段比较,血清中谷胱甘肽过氧化物酶和超氧化物歧化酶含量在生长阶段极显著高于育肥阶段(P<0.01)。血清触珠蛋白在不同品种及不同年龄阶段间均差异显著(P<0.05)。猪主要急性期蛋白在不同年龄阶段间存在明显差异(P<0.05),但品种间未出现显著性差异。老淮猪血清皮质醇水平极显著高于三元杂交猪(P<0.01)。

<正>本研究分析了杜长大三元杂交猪肌肉注射ACTH后2h内以及连续注射5天后血液中白细胞数目、常规生化指标以及皮质醇、T3、T4、胰岛素等激素,唾液中皮质醇的含量变化,探讨肌肉注射ACTH对生长猪代谢的影响。选用杜长大三元猪16头,8头注射ACTH,8头注射生理盐水,体重20 kg左右时安装颈静脉血管瘘管,恢复一周后进行ACTH肌肉注射实验,采集不同时间点