长江江豚监测现状及展望

长江江豚(Neophocaena asiaeorientalis)是唯一淡水生活的鼠海豚类,近年来种群数量严重下降,2013年被世界自然保护联盟红色名录列为“极度濒危”,2021年升级为我国国家一级重点保护野生动物,2022年种群有所恢复。长江江豚是长江生态系统健康的指示物种,我国科学家自20世纪50年代开始监测至今,监测方法不断更新完善,对长江江豚现状的了解也越来越充分。该文回顾了截线抽样法、水下被动声学及自动实时监测系统、无人机、环境DNA等监测方法的应用及取得的成效,分析监测方法存在的不足,预测技术发展趋势,提出改进建议,为长江江豚的监测及保护提供基本参考。     

环境DNA技术在环境监测与动物生产管理中的应用进展

概述了环境DNA技术目前的研究方向和分析原理,介绍了该技术在水生、土壤和植物生态系统中的应用进展,以及在监测畜舍环境、促进畜禽粪污无害化处理、检测动物营养与健康状况、调查动物群体遗传多样性、监测和预防畜禽疾病等动物生产管理中的应用前景。分析了环境DNA技术的优势与局限,并在此基础上提出了建立标准操作流程、提高检测精确度、积极推广应用环境DNA技术等建议。     

Characterization of microbial communities from coastal waters using microarrays

Molecular methods, including DNA probes, were used to identify and enumerate pathogenic Vibrio species in the Chesapeake Bay; our data indicated that Vibrio vulnificus exhibits seasonal fluctuations in number. Our work included a characterization of total microbial communities from the Bay; development of microarrays that identify and quantify the diversity of those communities; and observation of temporal changes in those communities. To identify members of the microbial community, we amplified the 16S rDNA gene from community DNA isolated from a biofilm sample collected from the Chesapeake Bay in February, 2000. The resultant 75 sequences were 95% or more similar to 7 species including two recently described Shewanella species, baltica and frigidimarina, that have not been previously isolated from the Chesapeake. When the genera of bacteria from biofilm after culturing are compared to those detected by subcloning amplified 16S fragments from community DNA, the cultured sample exhibited a strong bias. In oysters collected in February, the most common bacteria were previously unknown. Based on our 16S findings, we are developing microarrays to detect these and other microbial species in these estuarine communities. The microarrays will detect each species using four distinct loci, with the multiple loci serving as an internal control. The accuracy of the microarray will be measured using sentinel species such as Aeromonas species, Escherichia coli, and Vibrio vulnificus. Using microarrays, it should be possible to determine the annual fluctuations of bacterial species (culturable and non-culturable, pathogenic and non-pathogenic). The data may be applied to understanding patterns of environmental change; assessing the health of the Bay; and evaluating the risk of human illness associated with exposure to and ingestion of water and shellfish.     

大亚湾浮游细菌的变性梯度凝胶电泳分析

于2010年12月以及2011年6月采集了大亚湾9个站点的表层水样,利用PCR-变性梯度凝胶电泳(PCR-DGGE)技术对浮游细菌DNA指纹进行了分析.结果显示,大亚湾海域浮游细菌DNA指纹比较丰富,冬季和夏季样品中分别共得到32条和33条条带,每个样品得到的条带数为18~25条.DNA指纹的Shannon-Weaver多样性指数(H')和Pielou均匀度(J)的变化范围分别为2.48~2.88和0.80~0.93.DNA指纹条带数与温度和盐度明显正相关,而与p H、DO、透明度呈明显负相关关系.结果说明PCR-DGGE技术可以在一定程度上反映细菌的群落结构和多样性,但需要与其他技术相结合方能较全面反映细菌的结构与功能.     

Molecular prenatal diagnosis: the impact of modern technologies

Originally prenatal diagnosis was confined to the diagnosis of metabolic disorders and depended on assaying enzyme levels in amniotic fluid. With the development of recombinant DNA technology, molecular diagnosis became possible for some genetic conditions late in the 1970s. Here we briefly review the history of molecular prenatal diagnostic testing, using Duchenne muscular dystrophy as an example, and describe how over the last 30 years we have moved from offering testing to a few affected individuals using techniques, such as Southern blotting to identify deletions, to more rapid and accurate PCR-based testing which identifies the precise change in dystrophin for a greater number of families. We discuss the potential for safer, earlier prenatal genetic diagnosis using cell free fetal DNA in maternal blood before concluding by speculating on how more recent techniques, such as next generation sequencing, might further impact on the potential for molecular prenatal testing. Progress is not without its challenges, and as cytogenetics and molecular genetics begin to unite into one, we foresee the main challenge will not be in identifying the genetic change, but rather in interpreting its significance, particularly in the prenatal setting where we frequently have no phenotype on which to base interpretation. Copyright © 2010 John Wiley & Sons, Ltd.     

阴离子粘土(LDH)对DNA吸附行为的研究

以4种阴离子粘土做吸附剂,研究了阴离子粘土对DNA的吸附行为.同时,采用XRD、FTIR、UV-vis等表征手段对吸附前后的材料进行研究.吸附结果显示,二元阴离子粘土对DNA的吸附量高于三元阴离子粘土;3:1型阴离子粘土对DNA吸附力强于2:1型阴离子粘土.4种材料对DNA的吸附均符合Langmuir、Freundlich两种吸附等温模型,且Langmuir吸附等温模型拟合度更高,说明阴离子粘土对DNA的吸附为单层吸附.XRD结果显示,吸附前后阴离子粘土基本结构并未发生改变,晶形完好,层间距未有明显变化,表明阴离子粘土对DNA的吸附仅发生在表面,DNA并未进入阴离子粘土层间结构中.UV-vis及电泳结果显示,吸附前后DNA的构型并未发生改变,阴离子粘土的吸附并未对DNA产生较大的影响.     

一种活性污泥总DNA提取方法的优化

对3种活性污泥DNA提取方法———传统蛋白酶K-SDS-氯仿异戊醇法(CPSCI法)、液氮研磨法和脱腐处理法进行了对比,并针对CPSCI法从污泥量、保温时间、裂解方式及沉淀时间等4个方面进行了优化。结果表明,优化的蛋白酶K-氯仿-异戊醇法(OPSCI法)采用污泥量0.10 g、37℃静置10 min,加SDS常温旋涡振荡及异丙醇直接离心等条件可获得长度在23.1 kb左右的DNA,OD260nm/OD280nm为1.86,稀释10倍后即可进行16S rDNA PCR。该方法重复性好,提取得率高,纯度好,操作简便,为常规实验室开展活性污泥微生物多样性研究提供了帮助。     

Genotoxicity of ferric oxide nanoparticles in Raphanus sativus: Deciphering the role of signaling factors, oxidative stress and cell death

We have studied the genotoxic and apoptotic potential of ferric oxide nanoparticles(Fe_2O_3-NPs) in Raphanus sativus(radish).Fe_2O_3-NPs retarded the root length and seed germination in radish.Ultrathin sections of treated roots showed subcellular localization of Fe_2O_3-NPs,along with the appearance of damaged mitochondria and excessive vacuolization.Flow cytometric analysis of Fe_2O_3-NPs(1.0 mg/m L) treated groups exhibited 219.5%,161%,120.4% and 161.4% increase in intracellular reactive oxygen species(ROS),mitochondrial membrane potential(ΔΨm),nitric oxide(NO) and Ca2+influx in radish protoplasts.A concentration dependent increase in the antioxidative enzymes glutathione(GSH),catalase(CAT),superoxide dismutase(SOD) and lipid peroxidation(LPO) has been recorded.Comet assay showed a concentration dependent increase in deoxyribonucleic acid(DNA) strand breaks in Fe_2O_3-NPs treated groups.Cell cycle analysis revealed 88.4% of cells in sub-G1 apoptotic phase,suggesting cell death in Fe_2O_3-NPs(2.0 mg/m L) treated group.Taking together,the genotoxicity induced by Fe_2O_3-NPs highlights the importance of environmental risk associated with improper disposal of nanoparticles(NPs) and radish can serve as a good indicator for measuring the phytotoxicity of NPs grown in NP-polluted environment.     

水库水氯化消毒副产物毒性研究

为了证实水库出水的氯化消毒副产物CHClBr2和CHBr3具有一定的遗传毒性,将水库出水进行浓缩富集研究其对小鼠脾脏淋巴细胞DNA的影响。同时分别与水库枯水期和丰水期出水的小鼠彗星实验结果进行了对比,发现氯化消毒副产物CHClBr2和CHBr3的DNA损伤效应同时高于两个水期的出厂水。