4-氯-3-甲基苯酚(PCMC)暴露对成年斑马鱼内源性物质的影响

建立了4-氯-3-甲基苯酚（PCMC）的水生生物斑马鱼暴露毒理学模型,研究了斑马鱼PCMC暴露后内源性代谢物的变化.同时,在斑马鱼PCMC半数致死浓度LC₅₀的基础上,以1/10 LC₅₀为暴露浓度进行代谢组学分析.最后,经超高效液相色谱串联三重四级杆飞行时间质谱（UHPLC-Triple-TOF-MS）检测,结合主成分分析（PCA）和正交偏最小二乘法判别分析（OPLS-DA）进行代谢谱分析.结果发现,空白对照组和暴露组在代谢水平上有明显差异.进一步分析后筛选出9种上调物质和11种下调物质作为潜在生物标志物,主要包括氨基酸、嘌呤、糖类和脂质.根据差异代谢物生理功能分析发现,PCMC主要通过影响嘌呤代谢、甘油磷脂代谢、淀粉和蔗糖代谢、组氨酸代谢、丙酮酸代谢及苯丙氨酸、酪氨酸和色氨酸的生物合成,对斑马鱼的心血管系统、神经系统、蛋白代谢和能量代谢等方面产生毒性效应.本研究建立的代谢组学方法能全面评价氯酚类消毒剂对水生生物的毒性效应,为进一步阐明氯酚类消毒剂的毒性机制提供了理论基础.     

赤子爱胜蚓对不同浓度四溴双酚A代谢的急性与亚急性毒性响应

运用核磁共振一维氢谱研究不同浓度四溴双酚A(TBBPA)急性暴露与亚急性暴露对赤子爱胜蚓(Eisenia fetida)的代谢影响.通过设置不同剂量TBBPA进行急性暴露和亚急性暴露,运用正交偏最小二乘投影分析法(OPLS)比较急性和亚急性暴露下蚯蚓的代谢差异,研究蚯蚓代谢水平与暴露剂量的关系,以找出差异代谢物作为TBBPA毒性效应的代谢标志物.结果表明,急性暴露和亚急性暴露均能很好地区分不同剂量染毒组间的代谢差异,且TBBPA暴露剂量越大,蚯蚓体内代谢与空白组的差异越大.急性暴露和亚急性暴露后,亮氨酸、赖氨酸和甜菜碱等代谢物剂量随TBBPA剂量增加的变化趋势是一致的,亚急性暴露中的变化幅度大于急性暴露.TBBPA暴露剂量为100 mg·kg~(-1)时,赤子爱胜蚓受到毒害作用,体内代谢物变化显著,代谢平衡遭到破坏,差异代谢物对毒性效应的指示作用比较敏感.甜菜碱、亮氨酸和赖氨酸可作为标志性代谢物揭示TBBPA对赤子爱胜蚓在渗透压调节、能量供给以及免疫功能方面的毒性效应,为进一步探索TBBPA毒性机理提供理论基础.     

氯嘧磺隆降解菌的筛选及对污染土壤的生物修复

采用富集培养方法从江苏某激素研究所污水处理池排污口污泥中分离得到1株氯嘧磺隆降解菌,经菌株形态学特征和26S rDNA序列分析,鉴定为胶红酵母菌（Rhodotorula mucilaginosa ）.经降解条件优化,菌株对含100mg/L氯嘧磺隆的无机盐培养基中氯嘧磺隆的最佳降解条件为：接种量2.5%,培养温度28℃,pH 6.0,培养5d后降解率为87.33%;在氯嘧磺隆初始浓度为10mg/kg（干土）的模拟污染土壤中,菌株最佳降解条件为：接种量2.5%,温度25℃,pH 6.0,土壤含水量30%,静息培养30d后降解率为90.74%.土壤修复实验结果表明,施加胶红酵母菌后减轻了氯嘧磺隆对小麦幼苗的药害,在氯嘧磺隆浓度为10mg/kg的土壤中投加降解菌后,小麦的出苗率、株高、根长及鲜重均明显高于未投加降解菌的对照组（P < 0.05）.     

高密度聚乙烯微塑料与氯嘧磺隆对大豆生长和根际细菌群落的复合胁迫效应

随着我国农业的大力发展，塑料地膜和农药被广泛投入到农业生产中，而塑料地膜降解形成的微塑料和农药在土壤中累积也带来诸多环境问题.目前微塑料与农药单一作用的环境生物学效应已有报道，但两者复合胁迫对作物生长和根际土壤细菌群落的影响研究较少.因此，设计高密度聚乙烯微塑料（HDPE，500目）与磺酰脲类除草剂代表品种氯嘧磺隆共处理，研究其对大豆生长的影响，并通过高通量测序技术、互作网络和PICRUSt2功能分析，探究HDPE和氯嘧磺隆复合胁迫对大豆根际土壤细菌群落多样性、结构组成、菌群网络和土壤功能的影响，阐明HDPE和氯嘧磺隆对大豆的复合毒性.结果表明1% HDPE处理延长氯嘧磺隆在土壤中的半衰期（由11.5 d升至14.3 d），并且HDPE和氯嘧磺隆复合胁迫较单一污染物对大豆生长的影响更为明显.HiSeq 2500测序表明复合胁迫下的大豆根际细菌群落由20个门、312个属组成，门和属的组成数量显著少于对照和单一处理，并降低具有潜在生物防治特性、植物促生特性等功能菌属的相对丰度（如Nocardioides和Sphingomonas等）.Alpha多样性表明复合胁迫显著降低大豆根际细菌群落的丰富度与多样性，Beta多样性则表明复合胁迫显著改变大豆根际细菌群落结构.组间样品LEfSe和PICRUSt2功能分析表明复合胁迫调控根际细菌群落的优势菌群，并减弱土壤氨基酸代谢、能量代谢和脂质代谢等二级功能层的丰度占比.由属水平网络分析推测复合胁迫降低土壤细菌间的总连接数和网络密度，使网络结构简单化，维持网络稳定的重要菌群种类也发生变化.研究结果表明HDPE和氯嘧磺隆复合胁迫显著影响大豆生长，并改变大豆根际细菌群落结构、土壤功能和网络结构，相较于单一处理，复合胁迫的潜在危害更大.研究结果可为评价聚乙烯微塑料和氯嘧磺隆生态风险，以及污染土壤修复提供指导.     

三种磺酰脲类除草剂在土壤中的降解及吸附特性

采用室内模拟试验方法,研究了氟胺磺隆、氯吡嘧磺隆和磺酰磺隆3种磺酰脲类除草剂在5种不同种类土壤(江西红壤、太湖水稻土、东北黑土、南京黄棕壤、陕西潮土)中的降解及吸附特性.结果表明,这3种磺酰脲类除草剂在这5种不同土壤中的降解速率均比较快,且降解过程均遵循一级动力学方程,土壤pH值是影响土壤降解速率的主要因素.3种磺酰脲类除草剂在土壤中的吸附均符合Frendlich模型,且具有较低的吸附容量.3种磺酰脲类除草剂在不同土壤中的吸附自由能变化均小于40kJ.mol-1,吸附主要以物理吸附为主,吸附常数随土壤pH值的增大而逐渐减小.     

蚯蚓细胞色素P450亚酶及代谢组学对土壤亚致死剂量草甘膦除草剂的响应

以赤子爱胜蚓（Eisenia fetida）为受试生物,通过外源添加污染物的方法,研究了暴露于亚致死剂量草甘膦（添加剂量分别为5、10、20、40 mg·kg^-1）的土壤中7 d后,蚯蚓生长抑制率、细胞色素P450亚酶（CYP1A2、2C9与3A4）活力及代谢组学对土壤草甘膦的响应,旨在探讨亚致死剂量草甘膦能否对非靶标生物蚯蚓产生不良影响及其毒性作用机制,初步推断其毒性作用阈值,并筛选敏感生物标记物.结果表明：从蚯蚓生长抑制率来看,各处理组与对照组无显著差异;从蚯蚓CYP亚酶活力来看,在高剂量草甘膦（20、40 mg·kg^-1）处理下,CYP亚酶活力均受到显著抑制;基于代谢组学鉴定并筛选出14个小分子代谢物标记物,其中9个小分子代谢物（分别是1,6-二磷酸果糖、β-D-果糖6-磷酸、丙酮酸、组氨酸、赖氨酸、鸟氨酸、延胡索酸、肌酸、甜菜碱）在5~40 mg·kg^-1草甘膦暴露下显著低于对照水平,另外2个小分子代谢物（尿酸与二十-羟-二十烷四烯酸）则在草甘膦处理下显著升高.代谢组学的响应表明,草甘膦暴露（5~40 mg·kg^-1）致使蚯蚓糖酵解与三羧酸循环减弱,氨基酸代谢受损,嘌呤代谢紊乱,细胞渗透功能受损,对蚯蚓产生了明显毒副作用,其毒副作用还与细胞色素P450酶代谢相关.蚯蚓CYP亚酶活力与小分子代谢物标记物作为敏感生物标记物,可联合诊断土壤亚致死剂量草甘膦污染.     

污水地下渗滤系统微生物代谢组学分析

应用超高效液相色谱-质谱技术（UPLC-MS）,基于代谢组学方法,筛选不同进水有机负荷下地下渗滤系统潜在生物标志物.采用偏最小二乘法（PLS-DA）和主成分分析（PCA）模式识别方法对样品进行分型处理,根据模型的变量重要性因子（VIP值）筛选潜在生物标志物,分析代谢产物蕴含的生物学信息,研究代谢通路,通过RDA分析探索代谢产物与环境因子之间的相关性.PLS-DA模型结果表明,当进水有机负荷为250mg/L、400mg/L、500mg/L时,同一高度层代谢产物间存在显著性差异,共筛选出VIP值大于1.5的230种差异代谢物,以有机酸为主;同时也存在一些醇类、酚类等微生物代谢中间产物.此外,微生物代谢产物与模拟土柱高度有显著对应性.在水力负荷为0.14m³/（m²·d）,COD污染负荷为400mg/L条件下,3个高度层H2（500mm）、H4（1000mm）、H6（1500mm）筛选出53种VIP值大于1.5的差异代谢物,以酸类、酮类物质为主.RDA分析表明,随着有机负荷波动和剖面高度的变化,代谢产物受到氧化还原环境（ORP）与硝态氮（NO₃^-）的影响较大,呈负相关.     

基于PLS的火电二氧化硫减排因素分析研究

以四川省为例,以火电二氧化硫排放量作为因变量,对不同时间、不同地区的样本采样,采用偏最小二乘回归分析方法,建立自变量的偏最小二乘回归模型.该模型综合了多元回归分析、主成分分析及典型相关分析,很好地解决了由变量多重相关性、样本点少于自变量个数等引起的模型预测精度不高的问题,同时也避免了使用普通最小二乘回归方法所引起的模型回归系数异常的问题.结果显示,所建模型解释能力强、预测精度高,能为相关部门制定二氧化硫减排措施提供科学的依据.     

氟吡菌胺对斑马鱼的毒性效应

采用斑马鱼胚胎发育技术和传统毒理学方法, 研究了氟吡菌胺对斑马鱼胚胎、成鱼及仔鱼的毒性效应.结果发现,氟吡菌胺对3个阶段斑马鱼均具有致死能力,对仔鱼LC50(48h)值为0.204mg/L,成鱼LC50(96h)为0.286mg/L,幼鱼LC50(96h)为1.489mg/L.研究表明,0.0596mg/L以上浓度的氟吡菌胺对斑马鱼胚胎均有一定程度的致死效应,高浓度处理组胚胎出现心包囊肿、卵黄囊不吸收、黑色素沉积少及鱼体弯曲等症状;氟吡菌胺对斑马鱼成鱼进行14d暴露后,处理组成鱼体重下降,产卵量减少,有效卵量降低; 0.0298mg/L处理组144h子代仔鱼存活率虽然比较高,但存活鱼大部分出现心包囊肿、体弯曲等畸形.上述结果说明,水体中残留的氟吡菌胺对于斑马鱼各生命阶段的生长发育均具有潜在的危害.     

基于代谢组学指标的土壤亚致死剂量汞对蚯蚓的毒性研究

将赤子爱胜蚓(Eisenia fetida)暴露于亚致死剂量(1.0、5.0、25.0 mg·kg~(-1))汞污染土壤中4周,以蚯蚓个体(致死率、体重增长率)及小分子代谢物(代谢组)为指标研究其对汞的动态毒性响应,并采用最小二乘判别分析(OPLSA-DA)对暴露组及对照组的代谢物进行分类,进而识别潜在的标记物.结果表明,蚯蚓对汞的吸收尚未达到稳态,蚯蚓体内代谢物的响应依赖于暴露剂量及暴露时间.暴露1周时,蚯蚓体重略有增长但不显著;最低暴露剂量(1.0 mg·kg~(-1))导致蚯蚓体内亮氨酸、异亮氨酸、色氨酸、组氨酸、酪氨酸、5-氧脯氨酸、2-脱氧肌苷显著低于对照水平,柠檬酸、肌苷酸与腺苷在5.0、25.0 mg·kg~(-1)剂量下显著高于对照水平;暴露4周时,最高暴露剂量(25 mg·kg~(-1))显著抑制了蚯蚓的生长;汞添加组的蚯蚓体内谷氨酸、酪氨酸、马来酸、2-脱氧肌苷水平显著低于对照.上述代谢物对汞的动态变化表明它们可作为潜在生物标记物,用于诊断土壤汞污染.对代谢途径分析发现,1.0～25.0 mg·kg~(-1)汞即可破坏蚯蚓正常氨基酸代谢、三羧酸循环,扰乱能量代谢,对蚯蚓产生氧化损伤.本研究结果表明,相比个体水平的受试终点,代谢组学指标比个体水平指标能更敏感地响应较低剂量汞,是土壤汞污染生态毒性效应诊断的有效指标.另外,本研究结果可为土壤汞污染的风险评估及相关环境标准的修订提供大量基础数据.     

Relationship between metabolic enzyme activities and bioaccumulation kinetics of PAHs in zebrafish (Danio rerio)

Many studies have investigated bioaccumulation and metabolism of polycyclic aromatic hydrocarbons (PAHs) in aquatic organisms. However, lack of studies investigated both processes simultaneously, and the interaction between these two processes is less understood so far. This study investigated the bioaccumulation kinetics of PAHs and metabolic enzyme activities, including total cytochrome P450 (CYPs) and total superoxide dismutase (T-SOD), in zebrafish. Mature zebrafish was exposed to the mixture of phenanthrene and anthracene under constant concentration maintained by passive dosing systems for 16 days. The results showed that PAH concentrations in zebrafish experienced a peak value after exposure for 1.5 days, and then decreased gradually. The bioaccumulation equilibrium was achieved after exposure for 12 days. Both of the uptake rate constants (k_u) and the elimination rate constants (k_e) decreased after the peak value. The variation of PAH concentrations and metabolic enzyme activities in zebrafish had an interactive relationship. CYPs and T-SOD activities increased initially with the increase of PAH concentrations, but decreased to the lowest state when PAH concentrations reached the peak value. When the bioaccumulation equilibrium of PAHs was achieved, CYPs and T-SOD activities also reached the steady state. In general, CYPs and T-SOD activities were activated after exposure to PAHs. The decrease of PAH concentrations in zebrafish after the peak value may be attributed to the great drop of k_u and the variation of CYPs activities. This study suggests that an interactive relationship exists between bioaccumulation kinetics of PAHs and metabolic enzyme activities in aquatic organisms.     

Potential of capillary electrophoresis mass spectrometry for the characterization and monitoring of amine-derivatized naphthenic acids from oil sands process-affected water

Capillary electrophoresis coupled to mass spectrometry(CE–MS) was used for the analysis of naphthenic acid fraction compounds(NAFCs) of oil sands process-affected water(OSPW). A standard mixture of amine-derivatized naphthenic acids is injected directly onto the CE column and analyzed by CE–MS in less than 15 min. Time of flight MS analysis(TOFMS), optimized for high molecular weight ions, showed NAFCs between 250 and 800 m/z. With a quadrupole mass analyzer, only low-molecular weight NAFCs(between 100 and 450 m/z) are visible under our experimental conditions. Derivatization of NAFCs consisted of two-step amidation reactions mediated by 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide(EDC), or mediated by a mixture of EDC and N-hydroxysuccinimide, in dimethyl sulfoxide, dichloromethane or ethyl acetate. The optimum background electrolyte composition was determined to be 30%(V/V) methanol in water and 2%(V/V) formic acid. NAFCs extracted from OSPW in the Athabasca oil sands region were used to demonstrate the feasibility of CE–MS for the analysis of NAFCs in environmental samples, showing that the labeled naphthenic acids are in the mass range of 350 to 1500 m/z.     

Inactivation of Viruses and Bacteriophages as Models for Swine Hepatitis E Virus in Food Matrices

Hepatitis E virus has been recognised as a food-borne virus hazard in pork products, due to its zoonotic properties. This risk can be reduced by adequate treatment of the food to inactivate food-borne viruses. We used a spectrum of viruses and bacteriophages to evaluate the effect of three food treatments: high pressure processing (HPP), lactic acid (LA) and intense light pulse (ILP) treatments. On swine liver at 400 MPa for 10 min, HPP gave log₁₀ reductions of ≥4.2, ≥5.0 and 3.4 for feline calicivirus (FCV) 2280, FCV wildtype (wt) and murine norovirus 1 (MNV 1), respectively. Escherichia coli coliphage ?X174 displayed a lower reduction of 1.1, while Escherichia coli coliphage MS2 was unaffected. For ham at 600 MPa, the corresponding reductions were 4.1, 4.4, 2.9, 1.7 and 1.3 log₁₀. LA treatment at 2.2 M gave log₁₀ reductions in the viral spectrum of 0.29–2.1 for swine liver and 0.87–3.1 for ham, with ?X174 and MNV 1, respectively, as the most stable microorganisms. The ILP treatment gave log₁₀ reductions of 1.6–2.8 for swine liver, 0.97–2.2 for ham and 1.3–2.3 for sausage, at 15–60 J cm^?2, with MS2 as the most stable microorganism. The HPP treatment gave significantly (p < 0.05) greater virus reduction on swine liver than ham for the viruses at equivalent pressure/time combinations. For ILP treatment, reductions on swine liver were significantly (p < 0.05) greater than on ham for all microorganisms. The results presented here could be used in assessments of different strategies to protect consumers against virus contamination and in advice to food producers. Conservative model indicators for the pathogenic viruses could be suggested.     

利用GC/MS代谢组学技术检测纳米TiO2对多头绒泡菌代谢途径的扰动作用

纳米TiO_2(n TiO_2)具有广泛的应用价值.但n TiO_2在阳光或紫外线下具有很强的氧化作用,可对细胞产生强烈的毒害作用,因此其对生态环境的可能破坏作用值得关注.有关n TiO_2在暗环境中对生物的毒性作用报道还不多,还不能全面揭示n TiO_2的毒性作用机理.代谢组学分析是发现代谢差异、识别被干扰的代谢途径、了解n TiO_2毒性作用机理、评估n TiO_2细胞毒性的重要手段.本文采用GC/MS(气相色谱/质谱)技术检测了n TiO_2在暗环境下对多头绒泡菌原质团(单细胞)代谢组的影响,根据多变量识别模式分析发现了60余个被n TiO_2显著干扰的代谢物.这些代谢物涉及糖代谢、氨基酸代谢、核苷酸代谢、多胺生物合成、次生代谢途径等,为全面了解n TiO_2对细胞的毒性作用机理提供了有益补充.     

全国23个城市水源水中邻苯二甲酸酯代谢物浓度调查

采用UPLC-MS/MS方法对中国23个城市的90个自来水厂141个水源水样中5种常用PAEs的8种代谢产物进行检测.结果发现,所有自来水水源水中均检出了MPAEs,邻苯二甲酸单正丁酯（MnBP）检出浓度最高,为74.7ng/L.水源水中邻苯二甲酸单乙酯（MEP）,邻苯二甲酸单异丁酯（MiBP）和MnBP浓度与邻苯二甲酸二乙酯（DEP）,邻苯二甲酸二异丁酯（DiBP）和邻苯二甲酸二正丁酯（DnBP）浓度分别呈显著相关,表明两者可能是同源.邻苯二甲酸二（2-乙基己基）酯（DEHP）二级代谢产物所占DEHP一级、二级代谢产物浓度和（∑DEHP）为4.0%±5.6%,和天然水体中DEHP的微生物降解结果类似,水源水中的MPAEs可能来自PAEs在自然水体中的微生物降解.     

城市生活垃圾产量影响因素的路径分析——以厦门市为例

城市生活垃圾产量日益剧增,已成为制约我国城市健康和可持续发展的瓶颈.深入开展城市生活垃圾产生量的影响因素研究有助于揭示垃圾产生机制,把握其动态变化趋势,从而指导生活垃圾可持续管理及调控.因此,以厦门市为例,基于1979-2010年生活垃圾清运量及经济、生活水平、城市发展水平、社会人口学特征、自然等方面的时间序列数据,运用路径分析方法,开展了城市生活垃圾产量影响因素研究.结果表明,所构建的路径分析模型拟合优度很高,相比较简单的相关分析,很好地解释了因素与垃圾产量的因果关系;城市规模、人均可支配收入、城市化率、GDP的直接影响系数分别为0.58(p ＜0.001)、0.55(p＜0.001)、0.52(p ＜0.001)、0.51(p ＜0.001),均大于0.5,说明正影响作用很大;家庭特征直接影响系数为-0.40(p ＜0.001),为较大负影响;GDP通过旅游人次、城市规模通过家庭特征对垃圾产量有间接作用,影响系数分别为0.13(p ＜0.001)、0.31(p＜0.001);各因素总影响系数排序为城市规模(0.89,p＜0.001)＞ GDP(0.64,p ＜0.001)＞人均可支配收入(0.55,p ＜0.001)＞城市化率(0.52,p ＜0.001)＞家庭特征(-0.40,p＜0.001)＞旅游人次(0.19,p＜0.001)＞年均温度(0.09,p ＜0.001);旅游人次影响作用为中等偏弱,温度只引起生活垃圾产量的年内变动而不影响年际变化.     

Evaluation of joint toxicity of heavy metals and herbicide mixtures in soils to earthworms (Eisenia fetida)

It is widely acknowledged that a simplified and robust approach to evaluating thecombined effects of chemical mixtures is critical for ecological risk assessment (ERA) of contaminated soil. The earthworm (Eisenia fetida) was used as a model to study the combined effects of polymetallic contamination and the herbicide siduron in field soil using a microcosm experiment. The responses of multiple biomarkers, including the activities of catalase (CAT), superoxide dismutase (SOD), glutathione reductase (GR) and acetylcholine esterase (AChE), the concentrations of glycogen, soluble protein (SP), malonaldehyde (MDA), and metallothionein (MT), and the neutral red uptake test (NRU), were investigated. Multivariate analysis, Principal Component Analysis (PCA) and Spearman's Rank Correlations analysis (BVSTEP) revealed that the activities of AChE and CAT and the NRU content were the prognostic biomarkers capturing the minimum data set of all the variables. Internal Cd (tissue Cd) in earthworms was closely related to the health status of worms under combined contamination of heavy metals and siduron. The integrated effect (E_mix) calculated based on the activities of AChE and CAT and NRU content using the stress index method had significantly linear regression with internal Cd (p<0.01). E_mix(10), E_mix(20), and E_mix(50) were then calculated, at 1.27, 1.63 and 2.71 mg/kg dry weight, respectively. It could be concluded that a bioassay-based approach incorporating multivariate analysis and internal dose was pragmatic and applicable to evaluating combined effects of chemical mixtures in soils under the guidance of the top-down evaluation concept of combined toxicity.