Comparative analysis of receptor-binding specificity and pathogenicity in natural reassortant and non-reassortant H3N2 swine influenza virus

Genetic reassortment between human and avian influenza viruses can create pandemic viruses. Influenza surveillance of pigs in Jilin Province, in China during 2007–2008 revealed that there were two distinguishable genotypes: a human-like H3N2 genotype and a double-reassortant genotype derived from the human H3N2 and avian H5 viruses. In this study, viral infection potential, replication kinetics, and pathogenicity were compared. The solid-phase binding assay demonstrated that both viruses prominently maintained a preference for the human-type receptor and the reassortant A/swine/Jilin/37/2008 (Sw/JL/37/08) showed relatively higher binding affinities than the non-reassortant A/swine/Jilin/19/2007 (Sw/JL/19/07). Replication kinetics showed that Sw/JL/37/08 had higher replicability in MDCK cells than Sw/JL/19/07. The mouse experiments clearly revealed that Sw/JL/37/08 had higher virulence than Sw/JL/19/07 as measured by more significant body weight loss, higher viral lung load, delayed viral clearance from lungs, and more severe pulmonary lesions. Sequence analysis indicated that the absence of glycosylation sites at residue 126 of HA and 93 of NA, as well as the characteristic NS1 C-terminal PL residues of ESEV may account for the increased replication and pathogenicity of Sw/JL/37/08. These results may imply that human may have infection risk by the reassortant swine influenza virus and emphasize the necessity for enhanced viral surveillance strategies, which monitor reassortment events in nature to reduce the public health threat posed by influenza viruses with the potential for human-to-human transmission currently circulating in pig populations.     

猕猴属实验动物的动物福利要求

为了确保猕猴属实验动物的顺利出口,满足国际上对实验用猴越来越高的动物福利要求,本文从制度建设,人员要求,以及饲养、隔离、运输环节的动物福利要求进行了分析,起草了一套完整的动物福利规范,供国内养殖企业和检验检疫部门参考。     

抗A型产气荚膜梭菌α毒素单克隆抗体杂交瘤细胞系的建立与中和效应

用提取的 Ａ 型产气荚膜梭菌 α毒素包涵体免疫 Ｂ Ａ Ｌ Ｂ／ｃ 小鼠后, 取小鼠脾细胞与 Ｓ Ｐ２／０ 骨髓瘤细胞进行融合和克隆化,经间接 Ｅ Ｌ Ｉ Ｓ Ａ 筛选,共获得 １ Ａ８、１ Ｃ３、１ Ｄ５、１ Ｄ８、１ Ｆ１、１ Ｈ１ 和 ２ Ｅ３ ７ 株稳定分泌单克隆抗体（ Ｍ ｃ Ａｂ）的杂交瘤细胞株。经鉴定,７ 株 Ｍ ｃ Ａｂ 的 Ｉｇ 亚类有 Ｉｇ Ｇ１（１ Ｄ８）、 Ｉｇ Ｇ３（１ Ａ８、１ Ｃ３ 和 ２ Ｅ３）和 Ｉｇ Ｍ（１ Ｄ５、１ Ｆ１ 和 １ Ｈ１）。细胞培养上清和腹水抗体效价分别为 １∶５１２～１∶１ ０２４ 和 １∶１０６ ～１∶１０８ 。尤为重要的是,２ Ｅ３ 杂交瘤细胞株分泌的 Ｍ ｃ Ａｂ 不仅能够中和 α毒素的磷脂酶 Ｃ活性和溶血活性,而且能够对致死性腹腔感染小鼠产生良好的被动保护作用。     

猪蛔虫GST蛋白编码基因克隆及序列分析

为预测并分析猪蛔虫(Ascaris suum)谷胱甘肽S-转移酶(glutathione S-transferase,GST)蛋白编码基因的结构特征及其B细胞抗原表位,采用反转录-聚合酶链反应(RT-PCR)扩增猪蛔虫GST蛋白编码序列(coding sequence,CDS),将纯化的DNA与pMD19-T载体连接并转化入JM109感受态细胞,并将阳性菌液测序后进行序列分析、结构预测和抗原表位预测。结果表明,获得的猪蛔虫GST蛋白CDS序列与参考基因(Y10613.1)相比,在第243和248位核苷酸处发生变异,并导致第83位氨基酸由甲硫氨酸(Met)变异为苏氨酸(Thr);其优势B细胞抗原表位可能位于肽段29-38、43-47、80-87、114-120、129-131、143-145、190-194、201-204区域内或附近,其中最可能位于肽段43-46和201-204区域内或附近。研究结果为猪蛔虫基因工程疫苗的研发奠定了理论基础,同时为后续猪蛔虫GST基因功能和耐药相关性研究提供了基础资料。     

烟酸铬对西门塔尔牛瘤胃液pH值和氨态氮浓度的影响

试验选用4头平均体重(500±20)kg,年龄4岁,装有永久性瘤胃瘘管的西门塔尔牛阉牛,采用4×4拉丁方设计,以混合精料和风干玉米秸秆为基础日粮,研究日粮添加烟酸铬(0、40、801、20 mg/d)对瘤胃液pH值及氨态氮浓度的影响。结果表明:日粮添加烟酸铬对瘤胃pH值无显著差异(P>0.05);80 mg/d组瘤胃液氨态氮浓度显著低于对照组,其他各组间无明显差异(P>0.05)。综合以上结果,烟酸铬适宜添加水平为80 mg/d。     

Concurrent highly pathogenic porcine reproductive and respiratory syndrome virus infection accelerates Haemophilus parasuis infection in conventional pigs

This study was aimed at determining the effect of highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) on Haemophilus parasuis (HPS) in co-infection. A quantitative real-time PCR targeting infB gene, which is conserved among different HPS serotypes, was developed to improve the accuracy and speed of the detection of HPS. A total of 32 four-week-old conventional pigs were distributed randomly into four groups: pigs in group I were intranasally infected with HP-PRRSV first, and were then intraperitoneally inoculated with HPS on 5 days after HP-PRRSV infection; pigs in group II were intranasally inoculated with HP-PRRSV alone; pigs in group III were intraperitoneally inoculated with HPS alone; pigs in group IV were intraperitoneally inoculated with physiological saline. The amount of HPS in serum on 0, 3, 6, 9 and 12 days post-inoculation (dpi) with HPS were detected using the established quantitative real-time PCR. Clinical signs, pathological changes and histopathological lesions were observed. The amount of HPS in serum reached 10(6)copies/μl at 3 dpi with HPS in pigs of group I, while it arrived 10(5.7)copies/μl at 9 dpi with HPS in pigs of group III. The HPS loads in hearts and lungs were much higher than in other tissues. The study showed that HP-PRRSV was able to accelerate HPS infection and loads.     

Cdc2相关蛋白激酶PFTK1单克隆抗体的制备

本试验以Cdc2相关蛋白激酶PFTK1的120～141位多肽段与载体蛋白KLH偶联制备人工抗原,免疫Balb/c小鼠,取高免疫效价的小鼠脾细胞与骨髓瘤细胞SP2/0在PEG4000作用下进行融合,通过有限稀释法最终得到了2株分泌抗PFTK1抗体的杂交瘤细胞,分别命名为4E7和2E3,经鉴定2株抗体均为IgM亚型,轻链均为κ型。经过Western blotting鉴定该抗体能够与PFTK1(1～139肽段)特异性结合。此抗体经过优球蛋白沉淀法纯化后,其纯度达到86.8%。该抗体的制备为研究蛋白激酶PFTK1功能奠定了基础。     

骨外固定器在一例犬小腿骨骨折中的应用

骨外固定是指在骨折两端,经皮穿放钢针再用连接杆及钢针固定夹将裸露在皮外的针端彼此连结起来,从而固定骨折断端的一种治疗方法。论文通过对1例小腿骨折的德牧犬进行X光检查、手术及术后护理,探讨骨外固定在小动物临床上的应用,进一步提高对骨折的治疗水平。