蚕蛹蛋白水解技术研究进展

目前蚕业资源综合利用正作为一个新兴产业得到广泛关注。蚕蛹是缫丝过程中的副产物,富含蛋白质,含有人体需要的18种氨基酸,其中8种必需氨基酸含量达40%以上,其组成和比例非常符合WHO/FAO提出的氨基酸模式,是优质蛋白质及氨基酸来源。我国每年有10多万吨干蚕蛹可供利用,但大多被     

漳浦县中蜂高产措施

养中蜂最主要的目的是追求蜂蜜产量.目前国内有关中蜂技术较全面、系统的著作有杨冠煌老师的<中华蜜蜂>和龚凫羌先生的<中蜂饲养原理与方法>.但至今尚未见到有相当规模的示范蜂场的报道.我的技术措施在我地推广了20多年,现在已经在多个地区全面开花了. 今年春季的荔枝、龙眼花期,我县平均单脾产量为5～6.5千克,中蜂群产量在20～30千克,单脾平均产量为5.5～7千克.     

Immune responses and protective efficacy of a recombinant swinepox virus co-expressing HA1 genes of H3N2 and H1N1 swine influenza virus in mice and pigs

The recombinant swine poxvirus rSPV/H3-2A-H1 co-expressing HA1 genes of H3N2 and H1N1 subtype SIV has been constructed and identified. Inoculations of rSPV/H3-2A-H1 yielded ELISA and neutralization antibodies against SIV H1N1 and H3N2, and elicited potent H1N1 and H3N2 SIV-specific INF-γ response from T-lymphocytes in mice and pigs in this study. Complete protection against SIV H1N1 or H3N2 challenge in pigs was observed.     

海南坡鹿疾病发生的一般规律与防治对策

掌握海南坡鹿疾病发生与发展规律,有利于开展防治工作,促进种群恢复.根据多年的实践经验与观察,阐述了海南坡鹿普通疾病发生的一般规律:不同年龄的坡鹿个体多发性疾病存在差异;不同季节坡鹿的多发性疾病类型和发病率也不相同,并提出了相应的疾病防治措施与保护建议.     

山羊鼻内腺瘤和腺癌的病理学研究

发生于内蒙古自治区某地的山羊鼻内肿瘤呈地方性流行，临床上病羊初期流出大量浆液性鼻液，随后逐渐出现呼吸困难和鼻塞音，肿瘤可侵犯周围组织引起眼球突出和头颅变形，最后死于窒息。１４例山羊鼻内肿瘤的病理学研究表明，肿瘤起源于筛骨迷路粘膜的腺体（包括浆液腺和粘液腺），组织学上为腺瘤，以后可转变为腺癌，并向周围组织作浸润性生长，未见转移。２例作透射电镜检查见瘤组织有异型性，瘤组织和瘤组织内未见病毒样颗粒。发生     

氟对牛脾淋巴细胞膜ATP酶活性的影响

取健康牛脾脏,常规方法分离培养淋巴细胞,用不同浓度氟化钠（NaF）染毒24h。低渗溶胞法提取细胞膜,检测膜上ATP酶活性。结果显示,与对照组相比较,染氟组淋巴细胞膜ATP酶活性降低,组间差异显著或极显著。表明：氟能抑制牛脾淋巴细胞膜ATP酶活性。     

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.     

基于荧光显色的环介导等温扩增技术(LAMP)检测禽呼肠孤病毒

建立了一种基于荧光显色的禽呼肠孤病毒(ARV)环介导等温扩增(LAMP)检测方法.该检测方法使用针对ARV的p10基因8个区域的6条LAMP引物,能够在等温(63℃)条件下1h内完成反应,具有良好的敏感性和特异性,最低能够检出102个拷贝的病毒,敏感性比普通PCR高100倍,而对其他病毒检测结果为阴性.在荧光显色中分别应用SYBR Green Ⅰ和钙黄绿素作为显色剂,其结果与琼脂糖凝胶电泳一致.在对80份临床样本的检测中,使用LAMP和PCR检测出阳性样本的数量分别为68份和55份.因此,基于荧光显色的LAMP方法可以应用于ARV的快速检测,具有在科研机构、基层实验室特别是检测现场推广和应用的潜力.     

蜥蜴利什曼原虫在小鼠体内的分布及增殖研究

通过腹腔途径用体内含有荧光蛋白的蜥蜴利什曼原虫感染BALB/c小鼠,感染后采其脏器做冰冻切片,荧光染料染色后用荧光显微镜观察,并经PCR鉴定,结果显示蜥蜴利什曼原虫感染小鼠后,主要分布于心脏、肝脏、脾脏、肺脏、肾脏。另外,成功建立了利什曼原虫荧光定量PCR检测方法,并采用该方法检测感染蜥蜴利什曼原虫后BALB/c小鼠体内利什曼原虫的增殖情况,结果显示,感染13 d内,BALB/c小鼠体内蜥蜴利什曼原虫呈波浪状增殖。这一结果为研究蜥蜴利什曼原虫感染人和动物的致病机理和免疫方法及疫苗研制等方面提供了基础理论依据。     

PRRSV重组N蛋白表达、纯化及间接ELISA检测试剂盒的研制

采用RT-PCR扩增出大小409bp猪繁殖与呼吸综合征病毒核衣壳蛋白基因（PRRSV N gene）,将该基因克隆到表达载体pGEX-KG,构建重组表达载体pGEX-KG-N,转化表达菌株BL21（DM3）诱导表达,经SDS-PAGE和Western blot分析表明,重组N蛋白获得了高效表达,融合蛋白相对分子质量为41 000,并具有良好的免疫学活性。扩大诱导培养,收集菌体,提取包涵体,用GST-Protein Purtification Kit亲和层析纯化目的蛋白,SDS-PAGE电泳检测纯度达到90%以上,可满足诊断抗原质量要求。用纯化PRRSV重组核衣壳蛋白GST-N为包被抗原,建立检测PRRSV血清的间接ELISA诊断方法,并组装ELISA试剂盒。优化后抗原最适包被质量浓度为2.5mg/L;血清最适稀释度为1∶100;对血清样本检测临界值为0.224;与美国IDEXX公司的HerdChek ELISA试剂盒符合率为92%,特异性为95%,敏感性为90%;与猪细小病毒、猪伪狂犬病病毒、猪乙型脑炎病毒、猪圆环病毒、猪瘟病毒等常见猪繁殖障碍病标准阳性血清无交叉反应。ELISA试剂盒批内和批间变异系数分别为3.44%～6.34%和5.04%～7.64%。置4℃条件保存12个月,试剂盒稳定性无明显改变。该试剂盒对350份临床疑似血清检出率为88.6%。研制的ELISA试剂盒为PRRSV临床血清抗体检测及流行病学调查提供了技术手段。