试验方法和接种物来源对苯甲酸钠快速生物降解率的影响

化学品的快速生物降解性直接影响其在环境中的迁移、转化和归宿，是鉴别其环境危害性和持久性的基本指标，是政府管理部门对化学品进行风险管理的重要科学依据，在生态毒理测试中具有非常重要的地位。为探讨试验方法、接种物来源以及两者的交互效应对苯甲酸钠生物降解率的影响，2013~2015年间以广州地区2个主要处理生活污水的污水处理厂(DTS、LD)为接种物来源，采用OECD 301系列试验方法(301B、301D和301F)进行了苯甲酸钠的快速生物降解性试验，共获得89个标准化测试数据。统计分析结果显示，接种物来源、试验方法与接种物来源的交互效应对苯甲酸钠生物降解率影响不显著，但试验方法单一因素的影响显著。本研究表明，当受试物为易生物降解时，接种物来源、试验方法和接种物来源的交互效应对该受试物的生物降解率无显著的影响，但在同一条件下，选择不同的试验方法往往会导致最终生物降解率出现明显差异。     

污水处理厂接种物的多样性及对化学品生物降解的影响

生物降解是化学品在环境中的主要去除途径,接种物来源和特性是快速生物降解性测试结果产生差异的重要因素。从广州和贵阳市6家生活污水处理厂采集活性污泥和二级出水,分别培养得到64和52株细菌,共有菌属为气单胞菌(Aeromonas)、芽孢杆菌(Bacillus)和肠杆菌(Enterobacter)。变性梯度凝胶电泳结果显示,广州市3家污水处理厂活性污泥和二级出水条带数量分别为53~58和38~43条;而贵阳市3家污水处理厂分别为29~42和30~42条。6种来源的接种物对易降解的萘甲酸和难降解的4-硝基苄氯生物降解测试结果较一致,接种物来源差异对中等降解性的联苯生物降解结果影响显著,其接种物多样性与生物降解性呈正相关关系。     

化学品固有生物降解性试验OECD标准方法的适用性检验与改进

在经济合作与发展组织(Organization for Economic Co-operation and Development,OECD)颁布的3项固有生物降解标准测试方法中,改进的MITI(Ⅱ)(OECD 302C)方法因适用于难溶、易吸附和弱挥发等物质的固有生物降解性评估,成为目前适用范围较为广的方法。但OECD 302C对接种物规定特殊,需周期性采集不少于10个位点、不同类型的接种物进行驯养,采集、驯养和维持成本及操作复杂程度相对高,而OECD 302C方法获取的化学品生物降解率并未优于其他2项固有生物降解方法。因此,对10位点采集并驯养不同时间的接种物及污水处理厂曝气池采集的活性污泥进行微生物学特征比较,采用Biolog微生物鉴定系统及磷脂脂肪酸分析系统研究2种不同接种物的代谢活性及群落多样性,同时比较2种接种物对6种化学品的固有生物降解能力。结果表明,驯养接种物在驯养14 d后,活性与多样性开始下降,与真实环境来源接种物(10位点采集且未驯养接种物,即驯养0 d的接种物)的微生物群落差别较大。驯养1个月后的接种物在碳源利用活性、降解能力上与污水处理厂曝气池新鲜采集的活性污泥无明显差异。故笔者认为,在试验条件下,OECD 302C标准方法中的驯养接种物未达到预期目的,未经驯养的活性污泥更能反映真实环境,未来可以活性污泥作为该方法的接种物进行后续降解方法改进的研究。     

稳健统计在快速生物降解性试验质量分析中的应用

快速生物降解性试验在新化学物质的生态毒理测试中占据重要地位,其测试数据的可靠性直接影响后续的测试策略和物质的申报登记、以及新化学物质进入环境后的风险管理。为研究OECD 301系列试验(301B、301D和301F)的质量,选用本机构近5年来积累的标准化物质苯甲酸钠14 d快速生物降解率数据,首次采用稳健统计技术,以稳健总计统计量的Z比分数作为评价指标,对各试验数据和试验系统的质量开展评估。结果显示,当301B、301D和301F试验的苯甲酸钠降解率分别在63.8%~88.0%、63.4%~89.2%和66.0%~99.8%范围内时,表明测试结果满意,该批次试验数据质量良好;若苯甲酸钠降解率不在此范围,则表明测试结果可能存在问题,甚至得到的可能是错误数据,需要进一步验证。据此提出了对实际测试工作具有指导意义的质量分析对照表,以期为测试机构开展新化学物质快速生物降解性试验时的质量分析提供参考。     

BP-4的模拟生物降解特性研究

为研究紫外吸收剂2-羟基-4-甲氧基二苯甲酮-5-磺酸(BP-4)的最大生物降解能力,采用欧洲经济合作与发展组织(OECD)生物降解测试标准方法——303A模拟试验-污水好氧处理:活性污泥单元法,建立并进行了城市污水处理厂曝气池环境模拟试验。研究结果表明:在单一新鲜活性污泥作为接种物条件下,BP-4在第47天达到降解稳定期,溶解性有机碳(DOC)去除率平均值为36.90%±5.68%(n=18)。在以10种不同来源混合接种物条件下,BP-4在第41天达到降解稳定期,去除率平均值为33.11%±4.87%(n=17)。研究结果表明BP-4在环境中的降解能力有限,在本模拟方法试验条件下只能部分降解。     

球孢白僵菌胞外蛋白酶及类枯草杆菌蛋白酶的诱导

研究了接种物和诱导物对球孢白僵菌总胞外蛋白酶及类枯草杆菌蛋白酶（Ｐｒ１）产生的影响。结果表明，球孢白僵菌不同接种物产生的总胞外蛋白酶及Ｐｒ１活性有明显差异；不同诱导物对产酶水平也有较大影响。其中芽管状物在蝉蜕诱导液中的总胞外蛋白酶和Ｐｒ１活性均最高，诱导１２ｈ酶活性达高峰，不同来源的菌丝产酶水平有差异，在诱导液中，接种物总胞外蛋白酶和Ｐｒ１的变化趋势一致，但总胞外蛋白酶活性和Ｐｒ１活力间没有直接的     

农药米满(Tebufenozide)对淡水微型生物群落呼吸作用的影响

以淡水微型生物群落作为受试靶生物,研究了米满(Tebufenozide)对淡水微型生物群落呼吸作用的影响.用五天耗氧量抑制强度(RΔDO5)来表征农药对淡水微型生物群落呼吸作用的影响,并探讨了十二烷基苯磺酸钠、腐殖酸和硝酸盐等共存物质对米满生物效应的影响.本方法的表征参数可以推广到其它农药或化学品的环境安全评价.     

广州市某A2/O工艺城镇污水处理厂出水对大型溞的生殖毒性作用

城镇污水处理厂废水中存在大量生物毒性物质,但该废水对动物的生殖能力影响研究还十分缺乏。利用大型溞暴露实验,检测广州市某采用A~2/O工艺的城镇污水处理厂出水的急性毒性和生殖毒性。48 h急性毒性实验表明,所有水样均未表现出急性毒性效应。慢性生殖毒性(14 d)测试结果表明:(1)从进水到缺氧池出水均能显著提高第一胎产溞数量;(2)进水和沉砂池出水显著增加受试期总产溞数量;(3)从厌氧池到出水工艺段受试溞的第一胎产溞时间均推后;(4)好氧池出水的毒性显著降低,但仍然对大型溞具有生殖毒性。研究表明,好氧池(A~2/O)工艺能够显著去除具有大型溞生殖毒性的物质,但污水处理厂废水的生殖毒性仍需要引起关注。     

7种酚类化学物质对活性污泥的呼吸抑制作用

苯酚类化学物质是废水中常见的有机污染物,其对活性污泥的毒性数据对于污水处理厂稳定运行和化学品危害性评估具有重要意义。本研究采用活性污泥呼吸抑制试验(209)测定7种酚类化学物质对活性污泥的呼吸抑制作用。结果显示,2,6-二叔丁基苯酚和对特辛基苯酚对活性污泥未产生明显的毒性效应;2,4-二氯酚、2-苯基苯酚、4-硝基酚、4-氯酚和对甲酚等5种化学物质都对活性污泥呼吸有不同程度的抑制效应,3 h-EC50值分别为49.7、77.6、102、150.1和462 mg·L-1,构效关系分析结果表明-Cl、-NO2等官能团是导致活性污泥呼吸抑制效应增强的关键因素。在化学品生物降解性测试研究中,要确保有毒化学物质的测试浓度低于EC50值的1/10。     

城市污水厂污泥倾倒对海洋生物的毒性研究

以广州市和佛山市的5个城市污水处理厂的污泥为实验对象,选取费氏弧菌、蒙古裸腹溞、卤虫、裸项栉鰕虎鱼仔鱼4种不同营养级的海洋生物为受试对象,结合化学分析方法,研究广州区域周边污水污泥浸出液的生物毒性效应。结果显示各污水污泥浸出液的毒性均较大,且浸出液的Cu污染浓度较高;广州市污水污泥对4种受试生物的毒性效应要高于佛山市。从污泥毒性对受试生物的选择性方面分析,发现卤虫筛分能力强且灵敏度高,而费氏弧菌则相关性好且方法简捷。研究结果为反映城市污泥的生物毒性强度和选择合适毒性评价受试生物提供基础数据。     

Biodegradability of organic substances by ATP test

The aerobic biodegradability of organic substances in water is characterized by determination of energy change ‐ ATP content in microbial cells during biodegradation. A satisfactory result is obtained under the following conditions: the initial concentration of tested substance is 100mg/l (as DOC), the amount of the inoculum in the biological medium is 500mg/l (as MLSS), and the duration of test time is 14 days. The evaluating system (peak time, peak height and IA index) is proposed to assess the biodegradability of 46 organic compounds and 7 wastewaters.     

Characterization of the dissolved organic matter in sewage effluent of sequence batch reactor: the impact of carbon source

Dissolved organic matter (DOM) transformation in sequence batch reactor (SBR) fed with carbon sources of different biodegradability was investigated. During the biologic degradation process, the low molecular weight (MW) fraction (< 1 kDa) gradually decreased, while the refractory compounds with higher aromaticity were aggregated. Size exclusion chromatography (SEC) and fluorescence of excitation emission matrices (EEM) demonstrated that more biopolymers (polysaccharides or proteins) and humic-like substances were presented in the extracellular polymeric substance (EPS) extracted from the SBR fed with sodium acetate or glucose, while the EPS from SBR fed with slowly biodegradable dissolved organic carbon (DOC) substratestarch had relatively less biopolymers. Comparing the EfOM in sewage effluent of three SBRs, the effluent from SBR fed with starch is more aromatic. Organic carbon with MW>1 kDa as well as the hydrophobic fraction in DOM gradually increased with the carbon sources changing from sodium acetate to glucose and starch. The DOC fractionation and the EEM all demonstrated that EfOM from the effluent of the SBR fed with starch contained more fulvic acid-like substances comparing with the SBR fed with sodium acetate and glucose.     

有机化合物动态定量结构—生物降解关系（QSBR）模型研究

本文通过分析影响有机物生物降解的基本因素，指出ＱＳＢＲ模型应综合反映影响有机物生物降解的四个方面：取代化合物的摄入、取代化合物的诱导作用，毒性物质的形成和基础酶的缺乏，在此基础之上给出了动态ＱＳＢＲ的概念模型及方法学基础，并利用其研究了氯代芳香化合物的生物降解性。     

Estrogenic Effects of Effluents from Sewage Treatment Works

The occurrence of hermaphrodite fish in the lagoons of sewage treatment works led us to hypothesize that sewage effluent might contain a substance, or substances, estrogenic to fish. to test this hypothesis, we placed cages containing rainbow trout in the effluent from sewage-treatment works, and one to three weeks later measured the vitellogenin concentration in the plasma of the fish. Vitellogenin is a protein synthesized by the liver of oviparous fish in response to estradiol stimulation; it is then conveyed by the blood to the ovary, where it is sequestered by oocytes to form the yolk. Thus, the presence of vitellogenin in the plasma is indicative of estrogenic stimulation of the liver. an initial study, at a sewage-treatment works, showed that plasma vitellogenin concentrations rose rapidly and very markedly (over 1000-fold in three weeks) when trout were maintained in the effluent. an extensive nationwide survey was then conducted. Results were obtained from fifteen sewage-treatment works distributed throughout England. in all cases, exposure of trout to effluent resulted in a very pronounced increase (500 to 100,000-fold, depending on site) in the plasma vitellogenin concentration. Induction of vitellogenesis was also observed in carp, but to a much lesser extent than in trout.

The identity of the estrogenic substance is unknown. It is suggested that the two most likely possibilities are ethynylestradiol, originating from pharmaceutical use, or alkylphenol-ethoxylates (APE), originating from the biodegradation of surfactants and detergents during sewage treatment.

Laboratory studies on the potency of ethynylestradiol demonstrated that levels as low as 1 to 10 ng 1^-1 could generate the response shown by the caged fish and that positive responses may arise at 0.1 to 0.5 ng 1^-1. Further work is in progress on the potency of APE.     

Influence of environmental factors on biodegradation of quinalphos by <Emphasis Type="Italic">Bacillus thuringiensis</Emphasis>

Background

The extensive and intensive uses of organophosphorus insecticide—quinalphos in agriculture, pose a health hazard to animals, humans, and environment because of its persistence in the soil and crops. However, there is no much information available on the biodegradation of quinalphos by the soil micro-organisms, which play a significant role in detoxifying pesticides in the environment; so research is initiated in biodegradation of quinalphos.

Results

A soil bacterium strain, capable of utilizing quinalphos as its sole source of carbon and energy, was isolated from soil via the enrichment method on minimal salts medium (MSM). On the basis of morphological, biochemical and 16S rRNA gene sequence analysis, the bacterium was identified as to be Bacillus thuringiensis. Bacillus thuringiensis grew on quinalphos with a generation time of 28.38 min or 0.473 h in logarithmic phase. Maximum degradation of quinalphos was observed with an inoculum of 1.0 OD, an optimum pH (6.5–7.5), and an optimum temperature of 35–37 °C. Among the additional carbon and nitrogen sources, the carbon source—sodium acetate and nitrogen source—a yeast extract marginally improved the rate of degradation of quinalphos.

Conclusions

Display of degradation of quinalphos by B. thuringiensis in liquid culture in the present study indicates the potential of the culture for decontamination of quinalphos in polluted environment sites.

     

土壤和地下水中多环芳烃生物降解研究进展

多环芳烃是一类普遍存在于环境中的难降解的危险性"三致"有机污染物。受污染的土壤和地下水中的多环芳烃,生物降解是其归宿的主要途径。研究表明,对于土壤中低分子量多环芳烃类化合物,微生物一般以唯一碳源方式代谢;而大多数细菌和真菌对四环或四环以上的多环芳烃的降解作用一般以共代谢方式开始。文章重点论述了多环芳烃的来源、降解多环芳烃的微生物、生物降解机理、影响生物降解的因素以及生物修复方法。认为今后的研究方向是高分子量多环芳烃的降解机理与降解途径,基因工程技术在多环芳烃生物降解方面的应用,以及生物表面活性剂产生的机理及其在实际处理中的应用等。