RNA‑seq技术在水生生物生态毒理学中的应用进展

水域是地球环境的重要组成部分,也是最易受污染的生态系统之一。水生态系统中不同营养级别的水生生物可通过摄食、接触等多种途径摄入水体中的污染物。因此,监测水域污染物对水生生物和生态系统的影响,解析污染物对不同水生生物的毒性机制,筛选敏感、有效的生物标志物对生态毒理学研究和环境风险评价具有重要意义。RNA测序（RNA sequencing,RNA?seq）技术因所需样品量少,且不需参考序列,可在整体水平上鉴定基因差异表达,成为水生生物生态毒理学研究的最佳方法之一。基于此,介绍了RNA?seq技术的基本流程与数据分析过程,对该技术在不同生态位的水生生物（如鱼类、两栖类、贝类、甲壳类等）生态毒理学中的应用展开综述,并对RNA?seq技术面临的不足、挑战及发展趋势进行探讨,以期为该技术在水生生物生态毒理学研究中的应用,尤其是水生态环境中污染物胁迫水生生物机制的阐明及污染水域生态环境恢复提供参考。     

污染胁迫下的蚯蚓蛋白质组学研究进展

随着蛋白质组学的发展和每年有大量环境污染物进入土壤环境中,污染胁迫模式动物的相关生物标志物受到日益关注。蚯蚓,作为土壤中最大的无脊椎动物,是研究和评价土壤生态污染良好的模式动物。研究蚯蚓的蛋白质组学,对于寻找环境生态污染相关生物标志物和阐明生态毒理学机制有着十分重要的现实意义。目前已知的污染胁迫下蚯蚓蛋白质组学研究,提供了几个特定污染物胁迫蚯蚓的蛋白表达谱。这些蛋白涉及许多生物学过程,例如信号传导、糖酵解、能量代谢、分子伴侣和转录调节,提示了相关污染物可能的生态毒理学机制,有望成为潜在的生物标志物,用于有毒污染物的监测,但其特异性需要进一步试验的验证。对蚯蚓受污染胁迫的蛋白质组表达谱及潜在生物标志物进行简要综述。     

建立污染土壤修复标准的探讨

随着污染土壤修复技术的发展和污染土壤修复环境工程的实施,污染土壤修复标准的建立已成为检验污染土壤修复工程效果的瓶颈.我国现行的土壤环境质量标准已不能适应污染土壤修复效果评判的需要.本文在分析我国土壤环境质量标准不足的基础上,结合一些发达国家土壤修复标准以及我国土壤污染实际情况,提出建立污染土壤修复标准应从污染物的选择、分析检测方法、修复标准的分类、对地下水的保护和生态毒理学评价方面综合考虑.     

土壤污染的生态毒理诊断研究进展

土壤生态毒理学是污染土壤生态风险评价以及土壤污染控制的理论依据,土壤环境中有毒物质的生态毒理效应及其分子机制的研究是土壤生态毒理学的核心内容.土壤污染生态毒理诊断集合了土壤污染的全部信息,提供了土壤的整体毒性效应.因此,开展土壤污染生态毒理诊断研究有重要的理论意义和现实意义.本文在参考国内外土壤污染生态毒理诊断研究成果的基础上,介绍了常用土壤污染生态毒理效应诊断方法和指标,并探讨了不同方法的利弊.传统毒理诊断方法往往只能表征胁迫的程度,不能解释损伤和响应发生分子机理.污染土壤毒性评估的生物标记物和土壤污染分子诊断技术,可深入探讨外源胁迫下生物的解毒机制.本文对其进行了较为系统的介绍,并对未来的研究趋势进行展望.     

生物标志物（Biomarkers）在海洋环境监测中的研究与进展

污染物的存在给海洋环境和生态健康带来了巨大的压力,如何监测这些污染物并对其毒性进行科学的评价,成为当今环境科学关心的热点问题。生物标志物（Biomarkers）是毒理学研究的重要工具,它能对污染事件进行早期预警,并能在一定程度上评估生态风险。对近十多年来生物标志物的研究与应用进行了回顾,总结了不同水平（分子、细胞、个体、系统）生物标志物的种类、特征、检测方法和应用特点,并对现存挑战和未来发展趋向进行了展望,旨在系统的认识生物标志物,并为其在海洋环境监测中的合理应用提供理论借鉴。     

土壤低剂量芘污染对蚯蚓若干生化指标的影响

通过人工污染土壤的方法,设计芘的暴露浓度为0、60、120、240、480、960μg.kg-1.暴露实验进行1、3、7和14d后,分别检测蚯蚓内脏中细胞色素P450含量、谷胱甘肽转移酶(GST)、超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)活性和丙二醛(MDA)含量.结果表明,在供试浓度范围内,蚯蚓内脏中各生化指标对污染物暴露指示的敏感性存在差异:其中P450含量、GST和SOD活性最为敏感;POD和CAT活性次之;而MDA含量未对低剂量的芘暴露起到明显的指示作用.研究同时发现,低剂量污染物暴露的时间效应要强于剂量效应的影响.因而,在进行生态毒性诊断时,采用多指标和多时段的检测对增强指示的灵敏性和有效性尤为重要.     

基于双壳贝类指示的海洋微塑料污染监测与毒理学研究进展

微塑料(尺寸5 mm的塑料)是海洋环境中一种新型污染物,可被海洋生物误食并对其产生毒性效应。随着海洋微塑料污染化学与生态毒理学研究的深入,利用指示生物开展海洋微塑料污染监测及毒理学研究逐渐成为热点。海洋双壳贝类可通过滤食方式摄食环境中的微塑料,是海洋微塑料污染监测与毒理学研究理想的指示物种。本文综述了海洋双壳贝类体内微塑料的分析方法、不同海域双壳贝类体内微塑料的累积分布以及受控实验条件下微塑料对双壳贝类的生态毒理效应3个方面的研究进展。未来研究方向将主要包括:建立以双壳贝类为指示的海洋微塑料污染生物监测标准方法以及研究环境浓度微塑料对海洋双壳贝类的亚慢性毒性效应等。     

氮磷汇入驱动下的河流轮虫群落演替

【目的】解析污染条件下河流生物区系的演替过程及机制,可以为生物多样性减少提供诊断,为生物多样性保护和水生生态系统的生态安全提供科学支撑。【方法】以对污染具有指示作用的轮虫类群为研究对象,选择覆盖京津冀地区的、受到人类活动干扰的北三河水系为研究区域,分别对该区域富营养化相关水质参数和轮虫群落组成进行分析,在解析污染物排放导致的河流水体水质差异和轮虫群落组成差异的基础上,阐明导致轮虫群落演替的关键污染因子。【结果】水体水质指标的分析结果表明,西部河流与东部诸河具有明显不同的环境条件,其氮、磷指标(包括总氮、溶解性总氮、总磷、溶解性总磷及活性磷)浓度显著增加。轮虫群落的分析结果表明,与东部诸河相比,西部河流的物种数目、生物个体数、物种丰度及香农威纳指数均显著降低,且群落组成显著差异。群落-环境相关分析显示,总氮、溶解性总氮、总磷、溶解性总磷、活性磷以及氨氮是导致群落显著差异的关键环境因子。【结论】北三河水系西部河流的氮、磷含量显著高于东北部河流;在环境条件改变的前提下,北三河水系西部河流比东部河流物种丰度、生物量均显著减少,同时群落组成也显著改变,高耐污型物种出现。氮、磷元素的输入是导致轮虫群落演替的关键环境因子。     

四膜虫：毒理学与生态毒理学研究中的优良模式生物

四膜虫 (Tetrahymena)是毒理学与生态毒理学研究的良好模式生物之一。本文回顾了近年来以四膜虫为实验生物开展的毒理学与生态毒理学研究进展 ,并针对目前产生严重环境压力的持久性有毒化学污染物 ,提出了以四膜虫为模式生物开展相应的生态毒理学研究的前景以及可行的方法。     

沙蚕耐污染的特征及机理研究进展

沙蚕（Nereis diversicolor）是一种栖息于海陆交错带、具有重要生态学意义的无脊椎动物.近年来发现,沙蚕体内能够蓄积大量的重金属等有毒有害污染物,且表现出较强的污染耐性.文中就沙蚕对重金属和有机污染物具有的污染耐性特征和最新国内外研究进展进行了概述,并就其污染耐性形成的机理进行了分析.沙蚕的污染耐性机制可能是通过多种生态化学过程实现的,包括通过大量粘液的分泌,在沙蚕体表形成一层保护膜;与有效态以及溶解于水中的污染物离子结合,从而降低污染物的生物可利用性;加快排泄速率等.更多的研究认为,沙蚕将吸收进入体内的重金属等有毒有害污染物以无毒的形式存贮,达到解毒的目的.这些无毒形式主要包括:与金属结合蛋白MTs和MTLP的结合物,热稳定硫醇化合物（CHSTC）,或者与重金属相互结合形成不溶性的溶酶体/微粒和结石等颗粒物或沉淀物.最后,对沙蚕生态毒理学研究以及耐性机制研究的工作重点进行了展望.     

Ecotoxicological studies with the freshwater rotifer Brachionus calyciflorus

The use of the freshwater rotifer Brachionus calyciflorus as a test organism for ecotoxicological studies was examined. Changes in the feeding-, swimming behavior- and demographic characteristics of the rotifers exposed to increasing concentrations of copper (Cu) were studied and the relationships between the different test parameters and their ecological relevance evaluated.Relatively simple short-term bioassay methods were developed to measure the swimming activity (2 hours) and the filtration (5 hours) rate. Life-table experiments were performed to asses the long-term effects of Cu on the rotifers. Significant reductions in the filtration rate, the swimming activity and the demographic parameter r were observed at 20, 12 and 5 g 1^–1 of Cu, respectively. The implications and the possible ecotoxicological applications of the results are discussed.     

Use of swimming speed and egg ratio as predictors of the status of rotifer cultures in aquaculture

This study evaluated the use of egg ratio (eggs rotifer^–1) and swimming speed (mm min^–1) as prediction criteria for production and culture quality in mass cultures of the rotifer Brachionus plicatilis. Egg ratio was determined to be a suitable predictor of rotifer growth and production in the cultures. Low egg ratios (i.e., 0–0.17 eggs rotifer^–1) indicate reduced rotifer population over time (i.e., negative net population growth rates). However, at this time egg ratio dynamics are not suitably understood to predict in advance a sudden population collapse.Swimming speed of reproductive, egg-carrying females in the exponential growth phase was 40–45 mm min^–1. During exponential growth swimming speed was independent of the food used. Lower swimming speeds were obtained in late stationary phase (10–25 mm min^–1) when yeast was used as a food source. Both environmental factors (e.g., accumulating metabolites) and changes in nutritional state of the rotifers may have affected the swimming speed, but environmental factors appear to be the most important. We believe that swimming speed has the potential of becoming an accurate predictor of culture quality in mass cultures of rotifers.     

Rotifers in ecotoxicology: a review

In the past five years the use of rotifers in ecotoxicologial studies has substantially increased. This greater interest has been due to the central role of rotifers in freshwater planktonic communities, the ease and speed of making quantitative measurements of mortality and reproduction, their sensitivity to common pollutants, the commercial availability of cysts, and the existence of reliable, standardized protocols. The main endpoints used in ecotoxicology studies are reviewed, including mortality, reproduction, behavior, cellular biomarkers, mesocosms, and species diversity in natural populations. For each endpoint, published studies are cited, along with the compounds investigated, duration of exposure, and the LC50s, EC50s or NOECs reported. Rotifers have been included as part of a standardized mesocosm and in several large-scale, outdoor mesocosm studies. A critique of rotifer use in ecotoxicology is offered and it is concluded that the scientific basis for including rotifers as part of a battery of ecotoxicological tests is well established.     

Swimming speed and Reynolds numbers of eleven freshwater rotifer species

We obtained data on the swimming speed of 11 freshwater rotifer species. These data were analyzed in terms of Reynolds numbers given the fact that rotifers are in the evolutionary boundary between the use of cilia and swimming appendages. Swimming speed ranged from 0.174 to 0.542 mm ^–1. Philodina acuticornis odiosa was the fastest rotifer in terms of absolute speed. However, Gastropus hyptopus, the smallest of the rotifers analyzed, was the fastest (2.849) in terms of body-lengths s^–1. Reynolds numbers (Re) among the 11 species analyzed varied from 0.023 to 0.301. Lecane furcata had the smallest Re, while Epiphanes senta had the highest Re. The two bdelloid rotifers analyzed in this work swam five times faster than they crept. The importance of Reynolds numbers and drag coefficients are discussed in view of the present results and data found in the literature.     

珠江三角洲河网浮游轮虫的群落结构

通过2012年4次对珠江三角洲河网浮游动物的生态调查,研究该水域浮游轮虫的群落结构,包括种类组成、优势种、生物量及多样性等的时空分布,探讨了浮游轮虫群落结构与环境因子的相关关系,并阐析了浮游轮虫的聚群结构.结果表明： 共检出53种浮游轮虫.优势种类的季节演替及空间变化明显,针簇多肢轮虫占有较大优势.从季节变化来看,枯水期密度及生物量均大于丰水期,丰水期的生物多样性指数和均匀度要高于枯水期,浮游轮虫生物量和多样性的季节间差异显著;从空间分布来看,平均密度及平均生物量均呈现自西南向东北升高的趋势,最大密度及生物量均出现在市桥,而生物多样性指数和均匀度指数的空间变化趋势则相反,最高值均出现在青岐,浮游轮虫密度在各调查站位间差异显著,生物量及多样性在各调查站位间的差异不显著.浮游轮虫的密度与生物量、生物多样性指数与均匀度指数呈显著正相关,多样性与生物量呈显著负相关,生物多样性指数和均匀度指数随着生物量的增加显著降低.在不同季节,浮游轮虫密度与水温、pH、溶解氧、叶绿素a含量及总磷、总氮等环境因子密切相关.根据浮游轮虫密度对调查站位进行聚群分析得出5种聚群结构,说明相应调查站位的水质状况有显著差异.     

Red porgy (Pagrus pagrus) larval feeding performance and behavior at the onset of exogenous feeding

The feeding performance and behavior at the onset of exogenous feeding, 3 to 4 days after hatching (DAH), were studied in red porgy Pagrus pagrus larvae. Similar feeding efficiency and intensity were achieved for two feeding treatments (live or freeze-dried rotifers) suggesting that prey movement is not decisive for their detection and capture and demonstrating that at first feeding red porgy larvae can ingest inert food. Larvae feeding performance was not affected by a diet shift between treatments. Based on maximum rotifers consumption and gut evacuation time at 18 °C, the daily ration was estimated as 14.035 μg, considering 14 h of feeding and a 25% egg:female rotifer ratio. Larval swimming activity measured by video recording showed a close association with gut fullness and similar swimming patterns for 3 and 4 DAH larvae. However, 20.3% larger mouth gape and 54.6% higher swimming speed of the older larvae should provide a better feeding performance and more energy needed for growth.     

铜绿微囊藻对轮虫生命表参数和表型特征的影响

为评价铜绿微囊藻的有毒(Microcystin-producing Microcystis aeruginosa)、无毒(Microcystin-free M. aeruginosa)品系对轮虫种群增长和表型特征的影响, 研究探讨了萼花臂尾轮虫(Brachionus calyciflorus)在不同微囊藻溶液中的生活史参数及形态变化。实验中各处理组单位体积总含碳量为(20.61±0.15) g C/mL, 以使轮虫获得等碳量的食物供应。实验组轮虫分别用蛋白核小球藻(Chlorella pyrenoidosa)、斜生栅藻(Scenedesmus obliquus)、有毒和无毒微囊藻溶液单独投喂, 并用有毒、无毒蓝藻菌分别与不同绿藻的混合液投喂。生命表实验结果表明, 不同微囊藻混合液投喂的轮虫净生殖率R₀ (F=102.71, df=32, P<0.001)、世代时间T (F=17.05, df=32, P<0.001)和内禀增长率r_m(F=18.89, df=32, P<0.001)与对照组相比降低1.36%—210.34%。侧棘刺长(F=28.18, df=65, P<0.001)和游泳速度(F=181.69, df=65, P<0.001)下降2.63%—39.07%, 轮虫体长(F=690.04, df=65, P<0.001)变化显著。与绿藻投喂的轮虫参数值相比, 轮虫的生命表参数和表型特征变化规律随微囊藻溶液浓度改变。萼花臂尾轮虫受到铜绿微囊藻胁迫时, 生长繁殖受到抑制并通过改变自身形态以抵御不利的生存环境。     

Effects of three phthalate esters on the life-table demography of freshwater rotifer <Emphasis Type="Italic">Brachionus calyciflorus</Emphasis> Pallas

Phthalate esters (PAEs) are mainly used in the polymer industry as external plasticizers in PVC, and tend to migrate slowly out of the plastic, either into the air by volatilization or into water or other solvents by dissolution. Di-n-butyl phthalate (DBP), butyl benzyl phthalate (BBP) and di-2-ethylhexyl phthalate (DEHP) are three members of PAEs, identified as priority controlled hazardous substances by the United States Environmental Protection Agency, and have been shown to have potential for endocrine disrupting effects on vertebrates and humans. The effects of DBP, BBP and DEHP on survival and reproduction of the freshwater rotifer Brachionus calyciflorus were studied using life-table demographic methods. The results showed that all the life-table demographic parameters of B. calyciflorus were markedly affected by DBP and BBP, but not by DEHP. The net reproductive rate representing the output of reproduction was more affected than all the other parameters representing population growth, development or survival of the rotifers. Compared to the solvent control, DBP and BBP, both at 500 μg l⁻¹, significantly increased the net reproductive rate, and prolonged the generation time and the life expectancy at hatching of the rotifers. DBP at 50 μg l⁻¹ markedly decreased the intrinsic rate of population increase of the rotifers, but the reverse was true for BBP at 50 and 500 μg l⁻¹. Among all the parameters, the intrinsic rate of population increase was the most sensitive to DBP and BBP. The levels of PAEs in water from all the studied rivers and lakes in the world did not affect the population growth of rotifers.     

An experimental system for the automatic tracking and analysis of rotifer swimming behaviour

An automatic tracking system for rotifer swimming movements is described. The x and y coordinates of the Center of gravity of the animal are stored in the computer memory every twelfth of a second. Computer programs were developed to analyze the data and to calculate the average and the standard deviation for the following parameters per unit of time: distance traveled, turning angles toward the right and toward the left, distance from the point of departure as the crow flies. Histograms are traced by the computer showing the distribution of these values, as well as their evolution with time. This experimental system is dependable. The first values which we obtained for the swimming speed of rotifers correspond well with those which are given in the literature. The other parameters had not been measured until now.     

Regulation of rotifer densities by crustacean zooplankton in an oligotrophic montane lake in British Columbia

Summary To examine the relative demographic effects of predation and competition for food in rotifers during spring and summer in an oligotrophic lake, predator and competitor densities and food supplies were experimentally altered inside large enclosures. Abundances of rotifer species were positively correlated with experimental densities of fourth instar Chaoborus trivittatus larvae, a major crustacean predator in 1976. Experimental alteration of the densities of Daphnia rosea and Diaptomus leptopus and D. kenai in 1978 produced highly significant increases in rotifer biomass only under Daphnia removal, but not under copepod removal. Inorganic fertilizer additions to enclosures in 1978 and 1979 revealed minimal rotifer increases unless pulsed additions were large or Daphnia were also excluded. Large demographic reponses of rotifers to low fertilizer loadings in the absence of Daphnia confirmed the pre-eminence of competitive food limitation in producing rotifer scarcity in summer.