The Distribution of Bovine Trypanosomosis in Zimbabwe and an Evaluation of the Value of an Anti-trypanosomal Antibody Detection ELISA as a Tool for Monitoring the Effectiveness of Tsetse Control Operations

Tsetse have been cleared from large areas of Zimbabwe during the past 65 years. In most areas, they are prevented from re-invading cleared areas by barriers of odour-baited, insecticide-treated targets. A trypanosomosis survey was conducted to determine the effectiveness of such barriers against re-invasion and to confirm the absence of tsetse in areas where they had previously been eradicated. Parasitological diagnostic methods and an anti-trypanosomal antibody detection enzyme-linked immunosorbent assay (antibody ELISA) were used. The prevalence of trypanosomal infections in the tsetse-cleared areas was generally low. However, the prevalence of anti-trypanosomal antibodies was unexpectedly high in some areas. This high proportion of cattle with antibodies could, in most cases, be explained by recent or historic information on the distribution and density of tsetse. The results from the survey demonstrated the value of anti-trypanosomal antibody detection as an additional sensitive tool for monitoring the effectiveness of tsetse control operations.     

苹果茎沟病毒部分分离物的生物学特性与分子鉴定

通过昆诺藜接种鉴定和ELISA检测,从梨和苹果上分离获得苹果茎沟病毒(Apple stem grooving virus,ASGV)23个分离物.采用TC-RT-PCR对这些分离物进行扩增,均获得特异的扩增片段,PCR产物经5%PAGE电泳,出现大小约500、530和600bp的3种迁移率不同的泳动带型.根据PCR产物电泳迁移率的差异,选取3个来源于梨的分离物P-L4、P-6-1-17和P-3-2-67的PCR产物进行克隆与序列测定.经BLAST搜索,3个分离物的扩增片段与苹果分离物P-209的CP基因3′端核苷酸序列同源性分别为92.2%、90.4%和88.4%.3个分离物间的核苷酸序列也有较大差异,P-L4/P-6-1-17为95.5%、P-L4/P-3-2-67为90.4%、P-6-1-17/P-3-2-67为88.6%.     

动物组织中兽药残留免疫学检测结果的若干影响因素

从受检样品保存条件、样品处理方法及分析方法3个方面阐述了影响动物组织中兽药残留免疫学检测结果的因素.     

杆状病毒表达SARS冠状病毒纤突蛋白及其抗原性分析

本研究构建了SARS冠状病毒纤突蛋白(S)重组杆状病毒(rBac-SS).SDS-PAGE及Western-Blot分析表明约190Ku左右的重组SARS纤突蛋白(rSS)在rBac-SS感染圆昆虫细胞获得表达,并具有特异免疫反应原性.以rBac-SS感染的昆虫细胞裂解物稀释后直接包被ELISA板,与Vero细胞培养的全病毒裂解物比较,检测SARS-CoV康复病人血清特异抗体,表现出同样的敏感性和特异性;rBac-SS感染的昆虫细胞用于间接免疫荧光,快速检测血清特异抗体反应,具有良好的敏感性和特异性.结果显示,杆状病毒表达的rSS有望替代SARS-CoV全病毒,作为安全、敏感和特异的重组诊断抗原,并为探索重组亚单位疫苗的可行性奠定基础.     

Characterization of an endopolygalacturonase Gene cppg1 from Phytopathogenic Fungus Chondrostereum purpureum

Chondrostereum purpureum, a phytopathogenic fungus, produces endopolygalacturonase (endoPG) which has been suggested to have a causal role in the silver-leaf symptom of apple trees. In this paper, we detected C. purpureurn-derived endoPG at the infection sites using ELISA with a polyclonal antibody against endoPG I. A gene encoding endoPG I and its homolog were also isolated from the C. purpureum genome. The endoPG I gene was designated as cppg1. The cppg1 gene is the first fungal endoPG gene reported in the Basidiomycetes. Received 31 May 2000/ Accepted in revised form 13 September 2000     

The Use of a GUS Transformant of Trichoderma Harzianum, Strain T3a, to Study Metabolic Activity in the Spermosphere and Rhizosphere Related to Biocontrol of Pythium Damping-off And Root Rot

The activity of Trichoderma harzianum in the spermosphere and rhizosphere of different plant species was studied by use of a beta-glucuronidase (GUS) transformant (strain T3a). Hereby, direct observation of micro-habitats supporting metabolic activity of T. harzianum is reported. Germination of conidia and mycelial growth were not supported by exudates from healthy roots of various ages. Instead, growth and activity of T. harzianum depended on access to dead organic substrates such as seed coats, decaying roots, and wounds, including those caused by infecting pathogens. A correlation between the GUS activity of T. harzianum and the biomass of Pythium ultimum in infected roots was established. On the basis of our observations, we suggest that the biocontrol ability of T. harzianum involves competition with the pathogen for substrates including the seed coat, and wounded or infected root tissue.     

Evaluation of PCR,IEF and ELISA techniques for the detection and identification of potato cyst nematodes from field soil samples in England and Wales

Effective management of potato cyst nematodes (PCNs) requires simple, rapid and accurate identification and quantification of field populations. Soil samples from a survey of 484 fields in potato rotations in England and Wales were used to compare the identification and quantification of PCNs using IEF, PCR, ELISA and bait plant tests. The cyst counts and bait plant test revealed that 64.3% of field samples contained PCNs. Bait plant tests increased the detection rate of PCNs in field samples by 4–6.4%. This means that some infestations are cryptic and would not normally be detected by standard counts. IEF, PCR and ELISA methods distinguished between Globodera rostochiensis and G pallida and were able to register mixed populations; however they were not in full agreement. All methods suggested that G pallida is the dominant species in the field samples tested. The PCR results indicated that 66% of field samples contained pure G pallida, 8% contained pure G rostochiensis and 26% contained mixtures of the two species. Estimates of the relative process times taken per sample in the PCR, IEF and ELISA techniques are given. © 2001 Society of Chemical Industry     

棉花落叶型黄萎病菌的检测技术

采用Nit硝酸还原酶缺陷型突变体技术，共得到棉花黄萎病菌Nit突变体301个，其中A型155个，B型97个，C型29个。共检测到落叶型棉花黄萎病菌11株，除以前报道的6株外，发现了5个新菌株。针刺接种后，在棉株上均引起典型的落叶型症状。通过聚丙烯酰受凝胶电泳比较落叶型和非落叶型的8个代表菌株的蛋白质电泳图谱的差异，发现在浓度为7.5%的凝胶中，落叶型菌株有两条特异性的蛋白，其迁移率分别为Rf5=0.135,Rf9=0.405。用非落叶型菌株DF4、落叶型菌株SY12菌体的可溶性蛋白制备获得两个抗血清。用间接ELISA法检测，即能区分棉花黄萎病菌的立枯、枯萎、炭疽病菌，落叶型和非落叶型黄萎病菌均呈阳性反应，但落叶型黄萎病菌的OD值高于非落叶型黄萎病菌。     

梨中AVERMECTIN B1残留的酶联免疫吸附测定

本文建立了检测梨中Ａｖｅｒｍｅｃｔｉｎ Ｂ１的间接竞争酶联免疫吸附测定法（ＥＬＩＳＡ）。以结合物４″－Ｏ－（单）琥珀酰Ａｖｅｒｍｅｃｔｉｎ Ｂ１－ＢＳＡ免疫家兔，制备特异性针对Ａｖｅｒｍｅｃｔｉｎ Ｂ１的多克隆抗体。梨样本经甲醇提取后用ＥＬＩＳＡ检测。在６．０￣６０ｍｇ／ｋｇ添加浓度范围内，Ａｖｅｒｍｅｃｔｉｎ Ｂ１的回收率为８６％￣１０５％，变异系数（ＣＶ）为４％￣８％。样本检测限为０．１ｍｇ／     

从麦类种质资源中筛选大麦黄矮病毒(BYDV)抗原

用 ELISA 法鉴定了小麦近缘种赖草属(Leymus)、披碱草属(Elymus)、鹅冠草属(Roegneria)3个属的21个种,其中17个种抗 BYDV。21145份小麦品种中筛选到症状轻、病毒含量高的耐病品种忻县冬麦、江西早等29份。3604份大麦品种中筛选到症状轻、病毒含量低的抗病品种C13208、小麦近缘种(Agropyronintemedium)和普通小麦杂交的异源八倍体中4无芒,中5,远中7,陇远45、46,远中1001,忻4079以及附加系 L1。现已获得抗 BYDV 的以中4无芒、L1为亲本的杂交后代。