牛荚膜A型多杀性巴氏杆菌强菌株rpoE蛋白的原核表达及其免疫保护效力研究

为研究牛荚膜A型多杀性巴氏杆菌(PM)强菌株rpoE蛋白的免疫保护性,本研究利用PCR方法扩增牛荚膜A型PM的rpoE基因,构建pET-28a-rpoE重组表达质粒,经诱导表达后SDS-PAGE检测结果显示,表达的rpoE蛋白相对分子量约为20 ku,与理论值一致;western blot结果显示该蛋白具有很好的特异性和反应原性;纯化的rpoE蛋白经皮下免疫BALB/c小鼠,间接ELISA检测融合蛋白的免疫原性,结果显示rpoE蛋白可以诱导小鼠产生较高水平的特异性抗体;PM攻毒后,免疫组保护率为70%。本研究获得的牛荚膜A型PM重组rpoE蛋白具有良好的免疫原性,免疫小鼠后能够提供一定的保护,可以作为研发PM亚单位疫苗的候选抗原。     

Immune thrombocytopenia (ITP): Pathophysiology update and diagnostic dilemmas

Immune thrombocytopenia (ITP) is a common autoimmune bleeding disorder. The understanding of ITP pathogenesis is rapidly evolving. We now recognize ITP as a complex and heterogeneous syndrome that results from a combination of humoral and cell‐mediated attacks on platelets peripherally and megakaryocytes in the bone marrow. Autoantibody‐mediated ITP also varies in the pathway used to clear platelets, which depends on the platelet glycoprotein being targeted. Moreover, ITP patients present with variable bleeding severities and treatment responses that do not closely correlate with platelet count. A gold standard diagnostic test for ITP is lacking, and biomarkers to assess disease severity are in their infancy. This review provides an update on the immunopathogenesis of ITP and summarizes currently available tests for ITP diagnosis, prediction of disease severity, and treatment responses. Given the heterogeneous pathogenesis and clinical presentation of ITP, we highlight the need for the development of diagnostic and prognostic tests that would allow for the individualized management of a complex disease.     

Effects of different dietary sources and levels of selenium supplements on growth performance,antioxidant status and immune parameters in Ross 308 broiler chickens

1. Although different impacts of various sources of selenium (Se) on chicken performance have been largely studied, there is a lack of comparative experiments studying the effects of these sources on the immune system and antioxidant indices of broiler tissues. The aim of this study was to examine the effects of various sources and levels of dietary Se supplements on performance, antioxidant status and immune parameters in Ross 308 broiler chickens.

2. A total of 1200 1-d-old male broilers (Ross × Ross 308) were divided into 8 treatments with 6 replicate pens and 25 birds per pen. This experiment was conducted as a completely randomised design with a 4 × 2 factorial arrangement. Main factors included Se sources as sodium selenite (SS), Se-enriched yeast (SY), DL-selenomethionine (SM) and nano-selenium (NS) and levels at 0.1 or 0.4 mg/kg Se.

3. Dietary supplementation of organic Se sources significantly improved average daily gain (ADG), gain: feed ratio and European production efficiency factor (P < 0.05) compared to birds fed on diets supplemented with inorganic source. In addition, ADG was increased in response to increased level of supplemental Se. Based on contrast comparison, there were significant differences in these parameters between organic versus inorganic sources of Se. However, there was no difference between contrast comparisons of NS versus SM and SY.

4. Total anti-sheep red blood cell (SRBC) and Immunoglobulin G (IgG) titres and hypersensitivity were enhanced by increasing supplemental concentration of Se and using organic sources of Se rather than SS (P < 0.05).

5. Oxidation resistance assessment of tissues demonstrated that supplementation of organic sources of Se and increase in supplemental concentration of Se ameliorated glutathione peroxidase activity, total antioxidant capacity and malondialdehyde formation (P < 0.05). Mostly, there were significant differences between organic versus inorganic sources of Se in oxidation resistance.

6. Overall, dietary supplementation of 0.4 mg/kg Se from an organic source resulted in better production performance and immune system response. Moreover, minimum formation of malondialdehyde in broiler tissue was observed in birds fed on diets supplemented with SM at 0.4 mg/kg.

7. It can be concluded that SM is more effective than other sources of Se in reducing lipid oxidation.     

Marginal deficiencies of dietary arginine and methionine could suppress growth performance and immunological responses in broiler chickens

The present study was conducted to investigate the effects of different levels of dietary arginine (Arg) and methionine (Met) on performance, immune responses, and carcass characteristics of broiler chickens. A total of 540 day‐old Ross 308 broiler chicks were randomly assigned into the nine experimental diets, consisting five replicates of 12 birds each. Dietary treatments included three different levels (90%, 100%, and 110% of National Research Council [NRC] specifications) of either dietary Arg or Met, which were fed to the birds according to a 3 × 3 factorial arrangement of treatments during a 42 days feeding trial. Results showed that supplementation of Arg and Met into the deficient‐diets increased (p < .01) weight gains during all trial periods. Although average daily feed intake (ADFI) was not influenced by dietary treatments, increasing Arg up to 100% of NRC recommendations improved (p < .05) feed conversion ratio (FCR) throughout the trial period. Similarly, supplementation of deficient‐diets with Met improved FCR values. There was a significant (p < .01) Arg × Met interaction for ADFI during the starter period; increasing the dietary Arg level increased ADFI when the diets were deficient in Met, while had an opposite effect in diets containing higher dietary Met levels. On the other hand, dietary Met fortification improved (p = .067) FCR values to a greater extent in 110% Arg‐diets during the entire trial period. Although different levels of Arg and Met had no marked effects on carcass yield and abdominal fat percentage, supplemental Arg up to 100% of NRC values increased (p < .01) the relative weights of spleen and bursa of Fabricius. Furthermore, bursa weight was affected by Arg × Met interaction (p < .01), so that supplemental Arg level of 100% of NRC increased the relative bursa weight in birds that were fed diets containing 90% and 110% of Met. Serum uric acid level was decreased (p < .05) as a result of dietary Arg fortification up to 110% of NRC recommended values. Supplementation of deficient‐diets with Met decreased (p < .05) serum cholesterol level. Although Newcastle antibody titer was not affected by dietary Arg or Met levels, Arg fortification of deficient‐diets increased (p < .001) antibody responses against infectious bronchitis (IBV) and bursal (IBD) disease viruses. Similarly, Met supplementation of deficient‐diets increased IBD antibody titer. There were significant (p < .05) Arg × Met interactions for IBV and IBD titers; Met fortification of 110% Arg‐diets was more effective in increasing antibody titers. An increase in dietary Met level up to 100% of NRC values increased (p < .001) serum concentration of γ‐globulins. The present findings imply that supplemental Arg could affect feed efficiency and antibody responses when the diets were already fortified with a sufficient Met level.     

Change in intrarenal Ghrelin expression in immune complex-mediated glomerular disease in dogs

Ghrelin is a peptide hormone that is mainly produced by the stomach. The kidney is a major source of local ghrelin, and maintaining body fluid balance is considered a critical role of renal ghrelin. However, there are no reports on renal ghrelin in small animal medicine. The present study investigated the intrarenal localization of and change in ghrelin expression in dogs with immune complex-mediated glomerulonephritis (ICGN). Ghrelin immunoreactivity (IR) was observed in the distal tubules of normal kidneys. Ghrelin IR was weak in ICGN kidneys, and the quantitative ghrelin IR score was significantly lower in ICGN kidneys than in normal kidneys. In cases of ICGN, plasma creatinine concentrations showed a positive correlation with the ghrelin IR score.     

The protective effect of γ‐aminobutyric acid on the development of immune function in chickens under heat stress

This study aimed to investigate the protective effect of γ‐aminobutyric acid (GABA) on the development of immune function in chicks under heat stress (HS). One‐day‐old male Wenchang chicks were randomly divided into control (CK), HS and GABA+HS groups. The GABA+HS group was fed with 0.2 ml GABA solution (50 mg/kg) daily by oral gavage. The HS and GABA+HS groups were placed in 40 ± 0.5 °C environment for 2 h heat treatment from 13:00 each day. Blood samples were routinely taken at 14, 21, 28, 35 and 42 days respectively, and the contents of T and B lymphocyte subsets in the blood and tissue were analysed by flow cytometry after FITC/PE double staining; the plasma levels of interleukin (IL)‐2, immunoglobulin (Ig)A, IgG and IgM were determined using ELISA. The thymus and the bursa of fabricius were also collected to analyse for organ index and observe for the changes in tissue microstructure. In addition, the chicks received primary and secondary immunizations with attenuated Newcastle disease (ND) vaccine (LaSota strain) at 7 and 28 days respectively; conventional hemagglutination inhibition (HI) assay was performed to monitor the titre changes in plasma antibody against ND virus in the birds. Our results indicated that the indices of both thymus and bursa of fabricius, the intactness of tissue structure and development, the plasma levels of IL‐2, IgA, IgG and IgM, the titres of ND antibody, and the levels of B and T lymphocyte subsets in HS group were all significantly lower than those in CK group (p < 0.05). However, all above indices were significantly improved in GABA+HS group compared with those in HS group (p < 0.05). These results demonstrated that while HS seriously affected the development of immune function in Wenchang chicks, GABA effectively alleviated the damages of HS to the development of immune function in chicks.     

晶维他复合液体维生素对鸡理化性能的影响

本试验旨在研究复合维生素对鸡机体免疫能力、生长指标、新城疫抗体、血液生化指标、体尺性状的影响。试验选用1日龄鸡200只称重后随机分为5个处理组,每组40只,分别为A组(即42d全给药组)、B组(即隔天给药组)、C组(即前21d给药组)、D组(即后21d给药组)和对照E组(即42d全饮水组),试验周期为42d,E组自由饮水,A、B、C和D组在饮水中添加不同浓度的维生素。结果表明,B组、C组和D组鸡较A组、E组鸡的各项指标均有一定程度的提高,但是以B组效果较为明显,说明合理饲喂晶维他复合液体维生素可显著提高鸡的理化特性,但长期过量补充维生素会影响机体发育,降低机体的免疫力。     

2015年甘肃宁县口蹄疫、猪瘟、鸡新城疫免疫抗体检测与分析

为全面掌握甘肃宁县猪瘟、猪和羊 O型口蹄疫、鸡新城疫等主要动物疫病的免疫效果,对宁县18个乡镇共1795份畜禽血清进行了抗体检测。结果显示,2015年度3种主要动物疫病免疫抗体合格率除鸡新城疫外,其余均超过农业部规定标准。表明宁县2015年全县主要动物疫病防控工作总体效果良好。此次检测还表明,全县不同规模养殖场中,规模养殖场主要动物疫病抗体水平普遍好于散养户,说明规模化养殖有利于疫病的防控和净化。不同地域抗体检测结果表明,全县中区免疫效果最好,东区效果不佳,建议加强2015年东部地区的补免工作。     

防御素基因在乌金猪与约大乌猪组织中的表达及东亚飞蝗抗菌活性物质对防御素基因表达的调节作用

本试验旨在检测β防御素-1(p BD-1)、β防御素-2(p BD-2)和β防御素-3(p BD-3)基因在乌金猪和具有乌金猪血缘的约大乌猪不同组织中的表达,并探讨东亚飞蝗抗菌活性物质(LAAS)对猪胎儿皮肤成纤维细胞(PFSF)免疫应激参数和防御素(p BDs)基因表达的调节作用。采用实时荧光定量PCR方法检测2月龄乌金猪与约大乌猪心脏、肝脏、肺脏、肾脏、空肠、回肠、十二指肠、皮肤、肌肉、胰腺、睾丸或卵巢这12种组织中p BD-1、p BD-2和p BD-3基因的表达差异;采用培养至第4~5代的PFSF,通过脂多糖(LPS)诱导建立免疫应激模型,在DMEM/F12培养液中添加不同浓度(0、75、150、300、600和1 200μg/m L)的LAAS,考察LAAS对PFSF免疫应激参数和p BD-1、p BD-2、p BD-3基因表达的影响。结果表明:1)乌金猪血清溶菌酶(LSZ)活性、免疫球蛋白G(Ig G)含量显著高于约大乌猪(P0.05),血清一氧化氮(NO)含量极显著低于约大乌猪(P0.01)。2)约大乌猪大多数组织中p BD-1、p BD-2和p BD-3基因的表达量高于乌金猪;2个品种猪的p BD-1和p BD-3基因均在皮肤和卵巢中表达量较高,p BD-2基因在肝脏和肾脏中表达量较高。3)300μg/m L LAAS极显著降低了培养液中乳酸脱氢酶(LDH)活性(P0.01);LAAS极显著增加了培养液中NO含量(除75μg/m L组正常细胞外)(P0.01),正常细胞和应激细胞均在LAAS浓度为600μg/m L时达到最高;LAAS浓度为0、75和150μg/m L时应激细胞一氧化氮合酶(NOS)活性显著或极显著高于正常细胞(P0.05或P0.01);添加1 200μg/m L LAAS可极显著降低培养液中的LSZ活性(P0.01),LAAS浓度为150、300、600和1 200μg/m L时应激细胞与正常细胞中LSZ活性差异显著或极显著(P0.05或P0.01)。4)应激细胞p BD-1、p BD-2和p BD-3基因的表达量均高于正常细胞;当LAAS浓度为150μg/m L时,p BD-1、p BD-2和p BD-3基因的表达量最高。结果提示,乌金猪和约大乌猪防御素基因的表达存在明显的品种差异性和组织特异性,适宜浓度的LAAS能够降低PFSF中LDH活性,提高NO含量及NOS、LSZ活性,上调应激细胞与正常细胞防御素的表达量。     

断奶应激对幼龄反刍动物免疫系统的影响及其机理

断奶初期,幼龄反刍动物的消化和免疫系统等尚未发育完全,断奶应激会导致幼畜体内激素水平及免疫功能改变,引起免疫系统抑制,诱发炎症反应,阻碍幼畜生长发育,增加患病风险。本文从糖皮质激素、免疫细胞、急性期蛋白和相关细胞因子4个方面阐述了断奶应激对幼龄反刍动物免疫系统的影响及其机理,旨在为相关研究提供科学依据。