内蒙古部分地区骆驼和羊寄生蜱虫病毒组学分析

为了解内蒙古地区蜱虫病毒组学的本底数据,采用病毒宏基因组学方法对在内蒙古阿拉善盟左旗、右旗和四子王旗地区3个采样点采集骆驼和羊体表寄生的1789只蜱虫样品进行病毒宏基因组学分析,并对特定病毒进行巢式PCR扩增和测序,通过Clustal W和MEGA7.0等生物信息学软件对获得的病毒基因序列进行遗传进化分析.数据显示,蜱虫样品携带包括植物、脊椎动物和非脊椎动物等来源的17个病毒科和一些未分类的病毒;其中,2株弹状病毒具有丰富的遗传多样性,与新疆地区和长江地区的弹状病毒的同源性达到98.5％和96.26％,提示蜱虫弹状病毒可能是通过羊和骆驼等动物贸易导致了新疆和内蒙古地区,以及内地的跨区域传播;细小病毒仅在羊来源的蜱虫中检测到,与中国河北地区的山羊血清中的细小病毒形成同一进化分支,我们推测蜱虫细小病毒在国内不同地区间可跨区域传播,在进化分析过程中,发现这种病毒与多种的细小病毒的同源性都不低于50％,提示细小病毒可能具有遗传稳定性;Tamdy病毒与来自阿塞拜疆、乌兹别克斯坦和美国的Tamdy病毒均具有极高的同源性,结果显示该病毒在内蒙古地区已经出现,并存在潜在流行的可能,有必要对Tamdy病毒进行进一步的监测;在本研究中,我们鉴定的白蛉病毒与来自新疆的亚洲璃眼蜱所携带的博乐蜱虱病毒形成同一个进化分支,与新型布尼亚病毒和Heartland virus病毒的同源性达到50％以上,该结果提示,我们发现的蜱虫白蛉病毒可能具有潜在的致病性,需要对其流行情况和致病性进行监测和研究.本研究为完善内蒙古部分地区蜱虫病毒的多样性和本底情况提供了重要的基础数据.     

Epstein—Barr病毒载体及其在基因治疗中的应用

EB病毒具有多种生物学特性,据此构建了宿主范围广泛的EB病毒载体。因其较其它病毒载体具有明显优势,所以具有良好的应用前景,本文对EB病毒生物学特性和载体的研制及在基因治疗中的作用作了较全成阐述。     

HIV-1 Gag蛋白的高效表达调控研究

LTR位于HIV前病毒同DNA基因组的两末端,Gag基因位于5′端LTR的下游。LTR对转录的调控作用主要是由5′端LTR进行的。LTR具有启动子的作用,在结构上具有真核启动子的典型特征。另外,HIV-1 Gag蛋白还有能够自我装配成病毒样粒子(virus like particle,VLP)并从细胞中释放出来的重要特性。 我们利用痘苗病毒表达系统,探索了在细胞质环境中LTR中的多种功能结构对Gag蛋     

流式细胞仪检测纳帕海高原湿地浮游病毒和浮游细菌丰度

【背景】浮游病毒是水体微生物群落中重要的组成成分,深入研究浮游病毒的时空分布有助于更好地保护和开发当地的微生物资源。【目的】对采集到的纳帕海高原湿地水样中的浮游病毒和浮游细菌进行计数,揭示纳帕海高原湿地浮游病毒的分布规律。【方法】采用流式细胞仪检测2013年12月和2014年9月纳帕海高原湿地7个水样的浮游病毒与浮游细菌丰度,并对影响浮游病毒丰度的因素,如细菌丰度、叶绿素a含量以及其他环境因子进行了相关性分析。【结果】季节分布上,雨季浮游病毒和浮游细菌丰度高于旱季;水平分布上,原水样品的浮游病毒高于湿地水和淤泥水。旱季水样的浮游病毒丰度受到细菌丰度及叶绿素a浓度的影响较大;雨季水样的浮游病毒丰度受到水体的p H值和温度的影响较大。【结论】纳帕海高原湿地的浮游病毒和浮游细菌是比较活跃的。浮游病毒丰度在不同季节、不同采样点受到细菌丰度和叶绿素a浓度等因素的不同影响。在旱季噬菌体而非噬藻体或浮游植物病毒是纳帕海高原湿地中浮游病毒的优势种群。     

宏基因组测序分析东寨港红树林淤泥和水体微生物的多样性

为深入了解海南东寨港红树林生态系统微生物多样性及其在氮、磷、硫等代谢循环中的功能特点,本研究采用宏基因组测序,从物种注释与丰度、群落功能及多样性指数等角度,分析红树林淤泥和水体生境中微生物群落结构及生态功能的特异性。结果显示,在淤泥中检测到53个门、909个属的微生物类群,有3个占比超过1%的优势门类,其中变形杆菌门为83.78%,处于绝对优势,其下的12个优势属全部来自变形杆菌门;不动杆菌属是聚磷微生物的主要类群,其在淤泥中含量是水体的107.7倍,硫氧化单胞菌属、脱硫杆菌属是硫化物代谢的主要菌属,主要存在于淤泥生境当中。在水体中检测到64个门、1 522个属,包括13个优势门类、7个优势属;Nitrospinae和硝化螺旋菌门是亚硝酸盐氧化代谢的关键类群,两者在水体中占比分别是淤泥中的28.1倍和6.8倍。多样性评估得知,水体样品中的Shannon Wiener指数和Simpson指数均高于淤泥样品,两样品在属分类学单元上的Simpson指数趋近于1,表明红树林生态系统具有非常高的微生物多样性,水体生境的微生物多样性高于淤泥;亚硝酸盐的微生物代谢循环主要发生在水体生境中,微生物对磷的富集作用和硫化合物的氧化还原代谢主要发生在淤泥生境中。本研究有助于认识东寨港红树林湿地生境中的微生物资源状况,为保护红树林生态系统和开发利用其中的微生物资源提供依据。     

用于病毒研究的人源化小鼠研究进展简

人源化小鼠从起初的入-鼠嵌合体到目前具有人体免疫活性的模型不断演进,已用于人免疫缺陷病毒、EB病毒、丙型肝炎病毒和登革病毒等病原体的感染、发病机制和防治的研究,取得了很大进展。我们简要介绍几种有代表性的人源化小鼠模型及其在病毒研究中的应用。     

宏基因组学研究方法及其在病原体检测中的应用

在自然环境中存在着大量的微生物。这些微生物种群,尤其是许多新发性病毒,会对人和动物体的健康产生重大影响,而它们大多数都是不可培养的微生物。宏基因组学(metagenomics)的研究对象主要是非培养微生物。在特定环境中,它可有效地获取微生物种群的遗传组成、群落结构及生态作用等信息。随着高通量测序技术的快速发展,宏基因组学分析方法也已经应用到病毒学领域中,出现了病毒宏基因组学(viral metagenomics)。这一技术在新病毒鉴定等方面都取得了很多重要的研究成果。该综述主要对宏基因组学的发展历史、应用前景、生物信息学分析流程,及其在病毒鉴定中的应用进行简要介绍。     

用于病毒研究的人源化小鼠研究进展

人源化小鼠从起初的人-鼠嵌合体到目前具有人体免疫活性的模型不断演进,已用于人免疫缺陷病毒、EB病毒、丙型肝炎病毒和登革病毒等病原体的感染、发病机制和防治的研究,取得了很大进展。我们简要介绍几种有代表性的人源化小鼠模型及其在病毒研究中的应用。     

鸡咽食管癌的电子显微镜研究I．林县鸡咽食管癌中发现的类B型病毒颗粒和特殊包涵体

我们对新近发现的鸡咽食管癌进行了电镜研究。在瘤细胞浆内发现了非常独特的病毒包涵体,在瘤细胞浆和细胞间隙里发现了大小在70—90毫微米具有偏心核样物特征性成熟瘸毒颗粒。 因为它们的结构与小鼠乳腺癌的B型病毒非常相似,我们给它定名为类B型病毒颗粒。     

病毒介导的细胞融合:癌症发生和发展的新视点

细胞融合(cell fusion)具有重要的生理意义。然而,病毒介导的非生理性细胞融合可能促进癌症的发生和发展。相对于基因突变等细胞癌变的传统诱因,病毒的这种致癌机制能够更合理地解释我们在癌症中发现的许多现象。病毒介导的细胞融合可能诱导癌症的观点对我们重新认识病毒相关肿瘤发生发展的机制,并依此调整癌症治疗的策略具有重要意义。     

Elimination of human enteric viruses during conventional waste water treatment by activated sludge

The present study was undertaken to determine if viruses were selectively eliminated during waste water treatment. Human enteric viruses were detected at all steps of treatment in a conventional activated sludge waste water treatment plant. Liquid overlays and large volume sampling with multiple passages on BGM cells permitted the detection of poliovirus (serotypes 1, 2, and 3), coxsackievirus B (serotypes 1, 2, 3, 4, and 5), and echovirus (serotypes 3, 14, and 22), as well as reoviruses. The mean virus concentration was 95.1 most probable number of infectious units per litre (mpniu/L) in raw sewage, 23.3 in settled water, 1.4 in effluent after activated sludge treatment, and 40.3 mpniu/L in sludge samples. All samples of raw sewage and settled water, 79% of effluent water, and 94% of sludge samples contained viruses. The mean reduction was 75% after settling and 98% after activated sludge treatment. Poliovirus type 3 was rarely isolated after the activated sludge treatment, but was still detected in about one-third of the sludge samples. Reoviruses and coxsackieviruses were detected at similar rates from all samples and appear to be more resistant to the activated sludge treatment than poliovirus type 3. Poliovirus types 1 and 2 were present in almost every sample of raw sewage and settled water and still found in about half of the effluent and sludge samples, indicating a level of resistance similar to that of reoviruses and coxsackieviruses.     

Human pathogenic viruses at sewage sludge disposal sites in the Middle Atlantic region.

Human enteric viruses were detected in samples of water, crabs, and bottom sediments obtained from two sewage sludge disposal sites in the Atlantic Ocean. Viruses were isolated from sediments 17 months after the cessation of sludge dumping. These findings indicate that, under natural conditions, viruses can survive for a long period of time in the marine environment and that they may present potential public health problems to humans using these resources for food and recreation. The isolation of viruses in the absence of fecal indicator bacteria reinforces previous observations on the inadequacy of these bacteria for predicting the virological quality of water and shellfish.     

Human pathogenic viruses at sewage sludge disposal sites in the Middle Atlantic region

Human enteric viruses were detected in samples of water, crabs, and bottom sediments obtained from two sewage sludge disposal sites in the Atlantic Ocean. Viruses were isolated from sediments 17 months after the cessation of sludge dumping. These findings indicate that, under natural conditions, viruses can survive for a long period of time in the marine environment and that they may present potential public health problems to humans using these resources for food and recreation. The isolation of viruses in the absence of fecal indicator bacteria reinforces previous observations on the inadequacy of these bacteria for predicting the virological quality of water and shellfish.     

Survival of enteroviruses and coliform bacteria in a sludge lagoon.

Enteroviruses associate with aerobically and anaerobically digested sludge were determined before the addition of the sludge to a sludge lagoon. The fate of sludge-associated viruses was followed during detention of sludge in the lagoon and after application of sludge to land for disposal. While digested sludge was being added to the lagoon, enteroviruses were readily detected in grab samples of sludge from the lagoon. Sludge-associated viruses dropped to low or undetectable levels after disposal of sludge on land and during periods when addition of digested sludge to the lagoon was suspended. Changes in the levels of fecal coliforms in the lagooned sludge paralleled changes in the numbers of enteroviruses. Enteroviruses were not detected in water from deep wells located on the sludge disposal site or near the lagoon. During the initial part of the study, poliovirus serotypes accounted for greater than 90% of the viruses identified. Later, poliovirus serotypes comprised less than 40% of the virus isolates, and echoviruses and Coxsackieviruses were the most common enteroviruses identified.     

Inactivation of enteric viruses in wastewater sludge through dewatering by evaporation.

The effect of dewatering on the inactivation rates of enteric viruses in sludge was determined. For this study, water was evaporated from seeded raw sludge at 21 degrees C, and the loss of viral plaque-forming units was measured. Initial results with poliovirus showed that recoverable infectivity gradually decreased with the loss of water until the solids content reached about 65%. When the solids content was increased from 65 to 83%, a further, more dramatic decrease in virus titer of greater than three orders of magnitude was observed. This loss of infectivity was due to irreversible inactivation of poliovirus because viral particles were found to have released their RNA molecules which were extensively degraded. Viral inactivation in these experiments may have been at least partially caused by the evaporation process itself because similar effects on poliovirus particles were observed in distilled water after only partial loss of water by evaporation. Coxsackievirus and reovirus were also found to be inactivated in sludge under comparable conditions, which suggests that dewatering by evaporation may be a feasible method of inactivating all enteric viruses in sludge.     

Inactivation of enteric viruses in wastewater sludge through dewatering by evaporation.

The effect of dewatering on the inactivation rates of enteric viruses in sludge was determined. For this study, water was evaporated from seeded raw sludge at 21 degrees C, and the loss of viral plaque-forming units was measured. Initial results with poliovirus showed that recoverable infectivity gradually decreased with the loss of water until the solids content reached about 65%. When the solids content was increased from 65 to 83%, a further, more dramatic decrease in virus titer of greater than three orders of magnitude was observed. This loss of infectivity was due to irreversible inactivation of poliovirus because viral particles were found to have released their RNA molecules which were extensively degraded. Viral inactivation in these experiments may have been at least partially caused by the evaporation process itself because similar effects on poliovirus particles were observed in distilled water after only partial loss of water by evaporation. Coxsackievirus and reovirus were also found to be inactivated in sludge under comparable conditions, which suggests that dewatering by evaporation may be a feasible method of inactivating all enteric viruses in sludge.     

Effect of sludge type on poliovirus association with and recovery from sludge solids

Sludge type was found to affect the degree of association between seeded poliovirus type 1 (LSc) and sludge solids. The mean percent of solids-associated viruses for activated sludge mixed liquors, anaerobically digested sludges, and aerobically digested percent of solids-associated viruses for activated sludge mixed liquors, anaerobically digested sludges, and aerobically digested sludges was 57.2, 70.4, and 94.7, respectively. The degree of association between poliovirus and sludge solids was significantly greater for aerobically digested sludges than for the other two sludge types. Sludge solids associated viruses were eluted using 0.05 M glycine buffer, pH 10.5-11.0, and subsequently concentrated by organic flocculation. The effectiveness of the glycine method in the recovery of solids-associated viruses was also found to be affected by sludge type. Significantly lower mean poliovirus recovery was found for aerobically digested sludges (14.5%) than for mixed liquors or anaerobically digested sludges (72.3 and 60.2%, respectively). The eluent used in the method was not as effective in dissociating the virus from aerobic sludge solids as it was for the other two sludge types. All other virus adsorption-elution steps of the method (i.e., virus concentration steps) were equally effective in poliovirus recovery for all three sludge types. It is suggested that future methods developed for the recovery of viruses from sludges be evaluated for the various sludge types likely to be tested.     

Inactivation of indigenous viruses in raw sludge by air drying

Air drying of raw sludge caused inactivation of indigenous viruses. A gradual loss of infectivity occurred with the loss of water until the solids content reached about 80%. A more rapid decline of viral infectivity occurred with further dewatering.     

Inactivation of Indigenous Viruses in Raw Sludge by Air Drying

Air drying of raw sludge caused inactivation of indigenous viruses. A gradual loss of infectivity occurred with the loss of water until the solids content reached about 80%. A more rapid decline of viral infectivity occurred with further dewatering.     

Enteric viruses in the Danube river water and sludge.

A study of the viral pollution of the Roumanian course of the Danube river was carried out from 1972 to 1974. The presence of enteric viruses was investigated in 123 water samples, taken by the gauze swab method and in 116 sludge samples from the bottom of the river. The samples were collected from 20 sampling points along the river from Moldova Veche to Br?ila, in accordance with the water use and the main pollution sources. Viruses were recovered from 17.5% of the samples. The frequency of virus isolations from water samples was similar to that observed in sludge samples. Therefore, the viral pollution of the Romanian course of the Danube showed a low level in comparison with other rivers, though two concentration techniques into suckling mice and into tissue cultures, were used for each sample.