动物RNA病毒检测关键试剂—反转录酶(M-MLV RTase)的制备

反转录酶是动物RNA病毒分子诊断用到的关键试剂之一。为获得纯度高、活性高、制备简单经济的M-MLV RTase,本试验将M-MLV RTase的基因序列进行密码子优化,合成并构建表达载体。将表达载体转化到大肠杆菌E.coli Rosetta宿主菌种,经IPTG诱导后实现M-MLV RTase的可溶性表达。利用Ni2+柱亲和层析纯化载有6x His标记的重组MMLV RTase,对重组酶进行Western Blotting试验,鉴定为M-MLV RTase。通过对动物流感病毒RNA的普通RT-PCR和荧光RT-PCR检测来评价重组酶的活性并估测其活性单位。将活性单位相当的重组酶和商品酶进行比较,结果显示,制备的重组酶活性高,对不同浓度病毒RNA的检测结果与商品酶相当。研制的M-MLV RTase可以用于动物RNA病毒的分子诊断。     

Observations on pathogenic organisms in the airborne state

Summary Increasing numbers of human and animal diseases have been shown to be disseminated by micro-organisms in aerial suspension. The viability of such airborne organisms declines progressively as the result of physical and other factors which are summarized in the text. Relative humidity strongly influenced viability in experimental studies, but strain differences also appear to play an important role in certain diseases. Laboratory results were confirmed by observations made in the field on poliomyelitis, smallpox, foot and mouth disease, rinderpest and Newcastle disease; it is probable that Marek's disease, avian infectious bronchitis and numerous other maladies spread in the same manner. Long range dissemination by airstreams may be influenced, to some extent, by the virulence of the causal agent and by the fall-out rate for particles of a size which is optimal for deposition at the predilection site of the specific organism. Furthermore, local geographical and climatic factors may greatly modify distributional patterns of airborne disease during major epidemics or epizootics: specific outbreaks are cited as examples.

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Sumario Se ha demostrado que un número criente de enfermedades tanto en el hombre como en los animales son diseminadas por micro-organismos que se encuentran en suspension en el aire. La viabilidad de tales micro-organismos en el aire declina en forma progresiva como resultado de factores fisicos y otros factores sumarizados en el texto. La humedad relativa influencia de manera especial la viabilidad en estudios experimentales, pero las diferencias entre las cepas tambien parece jugar un importante rol en algunas enfermedades. Resultados de laboratorio fueron confirmados por observaciones hechas en el campo sobre poliomielitis, viruela, fiebre aftosa, peste bovina y enfermedad de Newcastle; Es probable que la enfermedad de Marek. la bronquitis infecciosa aviar y otras numerosas enfermedades seandiseminadas de la misma manera. La diseminación a grandes distancias por corrientes de aire puede ser influenciada, hasta cierto punto, por la virulencia del agente causal y por la tasa de acentamiento para particulas de un tama?o, las cuales son óptimas para su acentamiento en el lugar de predilección del organismo específico. Mas aun, factores locales geográficos y climáticos pueden modificar grandemente los patrones de distribución de enfermedades diseminadas por el aire durante grandes epidemias ó epizootias: Se citan como ejemplos brotes específicos.

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Résumé Il a été démontré qu'un nombre croissant de maladies humaine et animale sont dísséminées par des microorganismes en suspension dans l'atmosphère. La viabilité de tels agents pathogènes décroit progressivement en fonction de l'influence des facteurs physiques et autres qui sont énumérés dans ce texte. L'humidité relative influence fortement cette possibilité de survie comme l'ont montré des études expérimentales, mais des différences de souches semblent aussi jouer un r?le important pour certaines maladies. Les résultats expérimentaux ont été confirmés par des observations faites sur le terrain à propos de la poliomyélite, la variole, la fièvre aphteuse, la peste bovine et la maladie de Newcastle; il est probable que la maladie de Marek, la bronchite infectieuse aviaire et de nombreuses autres infections se répandent de la même manière. La dissémination à longue distance par les courants aériens peut être influencée, dans une certaine mesure, par la virulence de l'agent causal et par le pourcentage de particules dont la taille est optimale pour leur arrivée au site d'élection du microorganisme spécifique. En outre, des facteurs géographique et climatique locaux peuvent grandement modifier le schéma épidémiologique des maladies disséminées par voie aérienne; des foyers spécifiques sont cités en example.

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Based on a paper presented at a Meeting of the Central Canada Veterinary Association on 9th December 1970.     

检测CSFV、JEV、PRRSV三种RNA病毒多重RT-PCR方法的建立

猪瘟病毒(CSFV)、流行性乙型脑炎病毒(JEV)和猪繁殖与呼吸综合征病毒(PRRSV)是引起严重的种猪繁殖障碍的病原,而且经常混合感染,及时准确诊断是防治的前提。根据GenBank发表序列选取3对引物建立检测CSFV、JEV和PRRSV病毒的多重RT-PCR方法,扩增产物分别为508 bp、380 bp、263 bp。经与IDEXX商品化的检测CSW抗原试剂盒比较,二者的符合率为96.7%;扩增JEV和PRRSV PCR产物分别经EcoR V和Sau3A I酶切得到预期的片段。建立的多重RT-PCR检测JEV、PRRSV和CSFV敏感度分别为12.5个TCID_(50)、10个TCID_(50)和10~(-3)ng总RNA。结果表明该多重RT-PCR方法具有很好的特异性和敏感性,可用于临床三种病毒核酸的检测。     

疱疹病毒致病机制的研究进展

<正>疱疹病毒属于疱疹病毒科疱疹病毒属成员,是一类较大的双链DNA病毒,其感染宿主广泛,主要侵害皮肤、黏膜以及神经组织,严重影响着人及其他动物的健康。根据疱疹病毒的基因结构、同源性以及相关特点,可将其分为α、β、γ等3个亚科。α-疱疹病毒亚科包括人疱疹病毒1型(herps simplex virus 1,HSV-1)、水痘-带状疱疹病毒 (varicellazoster virus,VZV)、马立克病毒 (Marek's disease vi     

Observations on the pathogenic effects of Nematodirus helvetianus in dairy calves.

Six Holstein-Friesian calves 3 to 5 months old, were used in a small-scale experiment to determine the pathogenic effects of the thread-necked strongyle Nematodirus helvetianus. Two calves were kept as controls and 4 were infected, 2 with single doses of 25,000, and 2 with 50,000 N helvetianus larvae. Observations made during the 8 weeks of the experiment found that N helvetianus is capable of producing severe pathologic changes in the intestine of previously healthy calves. At 38 days after infection, the calves infected with 25,000 and 50,000 larvae gained 32.9% and 72.4% less weight respectively, than their controls. An increase in total lymphocytes was observed with no significant change in hemoglobin concentration. Adult worms were found to penetrate the intestinal mucosa with considerable destruction, including tunneling. Rectal temperatures were elevated in infected animals. The prepatent period for N helvetianus in single infection was 18 to 19 days. A spontaneous expulsion of the adult parasites was observed in 1 of the calves exposed to 50,000 larvae. Worms recovered in this animal numbered 565, whereas the number of worms recovered in another calf infected with 50,000 larvae was 21,860.     

饲料中动物源性成分的检测技术研究进展

痒病和牛海绵状脑病（疯牛病）都属于传染性海绵状脑病,其病原是蛋白质侵染颗粒——朊毒体（Prion）。痒病是羊的一种比较古老的疾病,而疯牛病是最早在英国出现的一种新病,1987年Weels等正式报道,然后迅速传播到欧洲多个国家和地区,给英国和欧洲的养牛业造成了巨大损失,并影响到相关产业。     

3种抗生素对3种常见鱼类病原菌的抑菌活性研究

为研究盐酸恩诺沙星、乳酸恩诺沙星和乳酸诺氟沙星对嗜水气单胞菌(Aeromonas hydrophila)、鳗弧菌(Vibrio anguillarum)和哈维氏弧菌(Vibrioharveyi)的体外抑菌活性,本实验通过测定抑菌圈直径、最小抑菌浓度(MIC)、最小杀菌浓度(MBC)和杀菌曲线,研究3种常用喹诺酮类抗生素药物对3种水产病原菌的体外药效学特性.结果表明:乳酸诺氟沙星对嗜水气单胞菌和哈维氏弧菌抑菌效果最好,MIC分别为0.244μg/mL和1.954μg/mL:盐酸恩诺沙星对鳗弧菌抑菌效果最好,MIC为0.122μg/mL;8 MIC的乳酸诺氟沙星药液在12 h内完全杀灭嗜水气单胞菌,在24 h内也能全部杀灭哈维氏弧菌,8 MIC的盐酸恩诺沙星药液在24 h内全部杀灭鳗弧菌.本研究为3种抗生素在水产养殖中的应用提供实验依据.     

动物饲料配方系统(AFS)软件研究

主要介绍《动物饲料配方系统》计算机软件的设计环境与工具、结构与内容、功能和使用方法，旨在为进一步推广应用该软件，促进饲料工业的发展，提高动物生产经济效益提供先进的技术。     

警惕动物病毒侵害人类

动物病毒侵害人类由来已久。艾滋病是已危及人类生存的全球性传染病,据世界卫生组织和联合国艾滋病规划处2004年11月23日公布的《艾滋病新疫情》报告中估计,2004年该病死亡310万人,HIV感染者总数为3940万人。现在许多科学家相信,该病是由灵长类动物病毒突变后,才成为广泛流行于人类的病毒。     

Gamma ray inactivation of some animal viruses.

Twenty samples of animal viruses comprising 14 different viruses in 12 families were subjected to varying doses of gamma irradiation from a 60Co source in a Gamma Cell 220 (Atomic Energy of Canada Limited) to determine lethal dose levels. The dose responses appeared linear throughout inactivation. The D10 values, that is the dose necessary to reduce infectivity by one log10, ranged from less than 0.20 Megarads to approximately 0.55 Megarads. There was not a complete inverse correlation between the target size (virion core) and the D10 value.     

饲料中致病性大肠杆菌的分离与鉴定

大肠杆菌可分为致病性大肠杆菌和非致病性大肠杆菌两类,二者在菌落形态、染色反应、培养特性和生化反应等方面无明显差异,但抗原构造不同.迄今已知大肠杆菌有菌体抗原(O)171种,表面抗原(K)103种,鞭毛抗原(H)60种,表现为许多血清型.在引起畜禽肠道疾病的血清型中有肠致病性大肠杆菌(EPEC)、肠产毒性大肠杆菌(ETEC)、肠侵袭性大肠杆菌(EIEC)和肠出血性大肠杆(EHEC)4种.致病性大肠杆菌主要通过畜禽的消化道感染,在畜禽中可引起多种综合征,包括羔羊和犊牛痢疾,马、绵羊流产,家禽败血症,慢性呼吸道疾病和输卵管炎,仔猪水肿病等.随着我国加入WTO和西部大开发计划的实施,我国的畜牧业生产进入了一个全新的发展阶段,各大、中、小饲料厂也相继不断发展和壮大,为了进一步了解青海省饲料中致病性大肠杆菌的分布情况,2004年3～4月,笔者等对西宁市某饲料厂的饲料随机抽样并进行了致病性大肠杆菌的分离与鉴定.现将结果报告如下.     

Observations on an enzyme-linked immunosorbent assay for the detection of antibodies against avian leukosis-sarcoma viruses

In the detection of antibodies against exogenous subgroup A avian leukosis viruses (ALVs) using a representative subgroup A virus, concordance between enzyme-linked immunosorbent assays (ELISAs) and serum neutralizations ranged from 83 to 95%. In ELISAs, subgroup A- and subgroup B-specific neutralizing antisera were equally reactive against ALVs of subgroups A, B, and E. Conversely, little cross-reactivity of high-titered subgroup E antisera was observed against subgroup A viruses. Significant cross-reactivities of spontaneously induced subgroup E-neutralizing antisera were observed when tested against a representative subgroup B ALV. Because some normal chickens spontaneously mount antibodies against infectious endogenous viruses, misleading results may be obtained if subgroup B or E ALVs are the source of target antigens in ELISAs.