Influence of amount and type of dietary fat on deposition, adipocyte count and iodine number of abdominal fat in broiler chickens

This study described the relation between the type and amount of dietary fat on the deposition of abdominal fat by broiler chickens. It was hypothesized that at higher fat intakes, the well-known lowering effect of polyunsaturated fatty acids on the deposition of abdominal fat would be diminished. Experimental diets were formulated to contain three levels of added fat (3%, 6% and 9%). Each level had different proportions of the saturated fatty acids (SFA) and unsaturated fatty acids (UFA) by installing the ratios of 1:1, 1:2, 1:3, 1:4 and 1:5 with the use of tallow and soybean oil. Arbor Acres chicks, aged 7 days, were fed one of the 15 experimental diets until they were aged 42 days. Feed and water were provided ad libitum. There was no systematic effect of the dietary fat type and the amount on the weight gain and the feed intake. The lowest SFA:UFA ratio of 1:5 produced the lowest feed conversion rates, irrespective of the amount of the fat in the diet. The abdominal fat deposition was similar in the birds fed on diets containing either 3% or 6% added fat, but deposition was lower than in those fed 9% fat. A decrease in the SFA:UFA ratio of the diet was associated with a dose-dependent decrease in abdominal fat, irrespective of the amount of fat in the diet. This observation leads to the rejection of the hypothesis stated above. A decrease in the dietary SFA:UFA from 1:1 to 1:4 caused a decrease in the number of the fat cells per surface unit of breast meat. It is concluded that an increased intake of soybean oil at the expense of tallow reduced the abdominal fat deposition and the number of fat cells in the breast meat of broiler chickens.     

天然草地放牧牦牛采食行为及食性选择的研究

对青南地区玉树县境内2牧场的草地植被及成年母牦牛的采食行为和选择性采食习性及其关系进行了探讨,结果表明：B牧场禁牧区的生物量显著高于A牧场的禁牧区及A,B牧场的放牧区（P〈0.05）,B牧场的可食植物量显著高于A牧场（P〈0.05）,A牧场的可食植物量只占地上生物量的一半以下;B牧场分别以小嵩草,矮嵩草,披碱草,针茅,蒲公英等优良牧草为优势种,次优势种,而A牧场以线叶嵩草为优势种,凤毛菊,狼毒,黄帚橐吾等毒杂草及早熟禾等为次优势种;B牧场以小嵩草、矮嵩草等莎草科和为主要采食植物,而在A牧场除以莎草科植物为主要采食植物外,由于可食牧草量的不足,还采食一些本不喜食的植物。矮嵩草和小嵩草是牦牛最喜食,选择性最高的植物;斑块内采食口数A牧场为50.6 bite/patch显著少于B牧场的81.1 bite/patch（P〈0.01）;斑块内采食站数A牧场为8.4 FS/patch显著多于B牧场的5.2 FS/patch（P〈0.001）;采食站内采食口数A牧场为6.6 bite/FS显著少于B牧场的17.4 bite/FS（P〈0.001）;采食速度A牧场为1.32 bites/sec显著慢于B牧场的1.46 bites/sec（P〈0.01）。放牧草地的食物资源的匮乏,使放牧牦牛的食物选择性降低,采食行为的内容变劣,A牧场草地处于严重退化状态;牦牛处于缺营养状态,草地的退化对牦牛的生产性能有很大的影响。     

Characterization of porcine circovirus type 2 (PCV2) infection in swine lymphocytes using mitogen-stimulated peripheral blood lymphocytes from healthy PCV2-carrier pigs

Information regarding the susceptibility of swine lymphocytes to PCV2 is rather limited. To further explore and characterize the PCV2 infection in swine lymphocytes, an in vitro model using concanavalin A (Con A)-stimulated peripheral blood lymphocytes (PBLs) obtained from clinically healthy PCV2-carrier pigs was introduced. It was found that the PCV2 antigen-containing rate was below 2% in PBLs from healthy PCV2-free pigs following treated simultaneously with Con A and PCV2. However, significantly higher PCV2 antigen- and nucleic acid-containing rates could be seen in Con A-stimulated PBLs from clinically healthy PCV2-carrier pigs. Prior to Con A treatment, both of the PCV2 antigen- and nucleic acid-containing rates in PBLs from healthy PCV2-carrier pigs were less than 1%; however, they reached 22.1+/-5.7% by flow cytometry and 27.1+/-6.5% by in situ hybridization, respectively, at 4-day post-incubation with Con A. Phenotyping of PCV2 antigen-containing cells revealed that PCV2-positive cells could be detected in both T and B lymphocyte populations within which IgM-positive B lymphocytes appeared to have a relatively higher positive rate. The Con A-stimulated PBLs also displayed a significantly higher viral load by the measurement of either PCV2 DNA copy number or viral titer when compared with the non-treated PBLs from healthy PCV2-carrier pigs. The results indicate that PBLs, especially IgM-bearing B lymphocytes, are indeed susceptible to PCV2 infection and PCV2 is capable of replicating in dividing lymphocytes. This activation-induced replication may explain in part the pathogenesis of lymphoid depletion in PMWS-affected pigs.     

Identification of bovine viral diarrhea virus type 1 in yaks (Bos poephagus grunniens) in the Himalayan region

Since cattle are widely infected by bovine viral diarrhea virus (BVDV) in India, we searched for pestivirus infection in yaks. Of 71 pure and crossbred yaks from Himalayan region, pestivirus antigen was detected by Ag-ELISA in three animals. Pestivirus in leukocyte and cell culture isolated virus samples originating from positive yaks was also confirmed by RT-PCR using panpestivirus specific primers selected from 5'-untranslated region (5' UTR). The 5' UTR, N(pro) and E2 regions were sequenced and used for genetic typing. Phylogenetic analysis revealed that pestiviruses detected in three Himalayan yaks were similar genetically, belonging to BVDV-1. Antigenic characterisation of yak pestivirus also confirmed the typing as BVDV-1. This is the first report on the identification of BVDV type 1 in yaks.     

基于Cap蛋白研制的猪圆环病毒2型疫苗研究进展

猪圆环病毒相关疾病(PCVAD)是危害世界养猪业的主要疫病之一,主要由猪圆环病毒2型(PCV-2)引起。该病毒可引起感染猪免疫系统的破坏,造成严重的免疫抑制,继而引发多种病毒/细菌的混合感染与继发感染,给世界养猪业造成了巨大经济损失。近年来疫苗免疫接种是防控PCVAD的有效手段,由于衣壳蛋白(capside,Cap)是PCV-2的主要免疫保护性抗原,因此常被用作研制PCV-2基因工程疫苗的理想靶抗原。论文就PCV-2Cap蛋白基因工程疫苗的研究进展进行综述,以期为今后PCV-2新型疫苗的研究提供参考。     

猪圆环病毒2d型的分子生物学特征和流行病学研究进展

猪圆环病毒2型(Porcine circovirus Type 2,PCV2)被认为是猪圆环病毒相关疾病(PCVAD)的主要病原体,分为多个基因型和亚型,具有很高的突变率,给猪场的疫病防控带来了巨大的挑战。虽然PCV2不同基因型间存在一定的交叉免疫保护作用,但是,在国内外多个接种了PCV2疫苗的猪群中,报道分离得到了突变型PCV2d毒株(又称PCV2b-1C,PCV2 mutant或m PCV2b)。为更好地防控PCV2d毒株的流行,文中就PCV2d的分子生物学特征和流行病学情况分别进行了综述。     

猪圆环病毒3型Cap蛋白单克隆抗体的制备及其在免疫组化试验中的初步应用

为制备猪圆环病毒3型(PCV3)Cap蛋白单克隆抗体(MAb),并初步应用于感染细胞或组织样品中PCV3抗原的间接免疫荧光试验(IFA)或免疫组化(IHC)检测,本研究将PCV3 Cap蛋白的去核定位信号肽基因克隆于原核表达载体pET-28a中,经IPTG诱导表达了具有免疫原性的融合蛋白。以表达的重组蛋白免疫BALB/c小鼠,共获得8株能够稳定分泌针对PCV3 Cap蛋白的MAb杂交瘤细胞株。Western blot与IFA试验结果显示,8株MAbs均能够与真核表达的重组Cap蛋白反应。选取3株杂交瘤细胞制备3株MAbs,小鼠腹水效价分别为1∶409 600、1∶204 800和1∶409 600。利用3株MAbs对自然感染PCV3临床病猪的腹股沟淋巴结进行IHC检测,结果显示3株MAbs的腹水均能够与感染PCV3的临床样品发生特异性的免疫反应。本研究制备的MAbs具有较好的特异性,为深入研究Cap蛋白的结构和PCV3快速诊断奠定了基础。     

猪腺病毒3型Protease蛋白在大肠杆菌中的表达及其抗体制备

试验旨在研究猪腺病毒3型（PADV3） Protease蛋白在大肠杆菌中的表达,并制备该蛋白的多克隆抗体。利用PCR扩增PADV3 Protease基因,构建重组原核表达载体pET28a-PADV3-Protease和真核表达载体pEGFP-PADV3-Protease,采用双酶切和测序鉴定;将原核表达载体pET28a-PADV3-Protease转化大肠杆菌BL21（DE3）感受态细胞得到重组原核表达菌株,经IPTG诱导收集蛋白,采用SDS-PAGE和Western blotting鉴定,将目的蛋白纯化后与佐剂乳化制备免疫原免疫家兔,制备Protease蛋白多克隆抗体;将真核表达载体pEGFP-PADV3-Protease转染HEK293细胞经G418筛选建立稳定表达EGFP-Protease融合蛋白的细胞系,以该稳定表达细胞系为包被抗原,采用免疫过氧化物酶单层细胞染色法（IPMA）检测抗体的免疫活性与抗体滴度。结果显示,Protease基因开放阅读框（ORF）为615 bp,原核表达系统中Protease蛋白以包涵体形式存在,分子质量大小为23 ku,与真核细胞表达的Protease蛋白分子质量一致,该蛋白在细胞核与细胞质中均有分布,制备的Protease蛋白多克隆抗体能与EGFP-Protease融合蛋白稳定表达真核细胞系发生特异性的反应,与对照细胞无反应。本试验构建了Protease蛋白原核表达菌株和真核表达细胞株,制备的PADV3-Protease蛋白多克隆抗体免疫活性良好,为进一步研究Protease蛋白的生物学功能和PADV3的血清学诊断提供了基础材料。     

2013~2017年江西省猪圆环病毒2型分子流行病学调查

为掌握江西地区猪圆环病毒2型（PCV2）的分子流行病学及其遗传变异情况,本研究运用实验室已建立的PCR方法对2013~2017年采集自江西省南昌市、宜春市、新余市、赣州市、吉安市、九江市、上饶市、抚州市、景德镇市、鹰潭市等10个地区的1 082份疑似PCV2感染猪病料组织进行病原学检测,并通过PCR扩增、克隆和基因测序对PCV2的全基因组序列进行分析。研究表明,1 082份病料中有610份为PCV2阳性,总阳性率为56.4%,其中2013~2017年阳性率分别为52.7%、48.8%、58.5%、71.0%和67.4%。共获得89株PCV2全基因组序列,其中有83株为PCV2b基因型,其中53株归类于PCV2b-1C基因亚群,30株归为PCV2b-1A/1B基因亚群;6株为PCV2a型。测序毒株与参考毒株ORF2基因核苷酸、氨基酸序列同源性分别为88.9%~100.0%和86.0%~100.0%;氨基酸序列分析表明,Cap蛋白存在20个氨基酸突变位点。本研究表明,江西地区猪群中PCV2的主导基因型为PCV2b,其中又以PCV2b-1C基因亚群占据主导地位,同时也存在少量PCV2a基因型。