Detection of antibiotic resistance and tetracycline resistance genes in Enterobacteriaceae isolated from the Pearl rivers in South China

This study investigated antibiotic resistance profiles and tetracycline resistance genes in Enterobacteriaceae family isolates from the Pearl rivers. The Enterobacteriaceae isolates were tested for susceptibility to seven antibiotics ampicillin, chloramphenicol, ciprofloxacin, levofloxacin, sulphamethoxazole/trimethoprim, tetracycline and trimethoprim. In Liuxi reservoir, with an exception to ampicillin resistant strains (11%) no other antibiotic resistance bacterial strains were detected. However, multiple drug resistance in bacterial isolates from the other sites of Pearl rivers was observed which is possibly due to sewage discharge and input from other anthropogenic sources along the rivers. Four tetracycline resistance genes tet A, tet B, tet C and tet D were detected in the isolates from the rivers. The genes tet A and tet B were widely detected with the detection frequencies of 43% and 40% respectively. Ciprofloxacin and levofloxacin resistant enteric bacteria were also isolated from the pig and duck manures which suggest a wider distribution of human specific drugs in the environment. This investigation provided a baseline data on antibiotic resistance profiles and tetracycline resistance genes in the Pearl rivers delta.     

油田污染土壤中石油烃类微生物及其降解基因的分布

以原油为唯一碳源,运用传统分离培养法,从中国江苏省淮安市金南油田石油污染土壤中筛选出9株具有原油降解功能的菌株并分别命名为JN1~JN9。对其进行形态观察、生理生化实验和分子生物学鉴定,最终确定其分别隶属于Lysinibacillus、Pseudomonas、Raoultella等5个菌属;在适宜条件下进行降解实验,9株菌对原油的降解率最高可达到32.63%;进一步对9株菌进行质粒和基因组上原油降解基因的分析,结果表明,GSTs基因广泛分布在其中8株菌的基因组上,同时JN1、JN3、JN4的质粒上也检测到了GSTs基因; alkB基因分别在7株菌的基因组中和2株菌的质粒检出;LmPH基因仅在JN9的基因组上检出。利用分子生物学手段对石油降解菌降解基因的分布的初步研究结果,可为原油污染场地的微生物修复提供理论依据。     

堆肥对污泥中四环素类抗生素及抗性基因的影响

近年来,抗生素及抗性基因的污染与扩散成为全球关注的热点。污泥,作为污水处理系统的副产物,是环境中抗生素抗性基因的重要存储库。污泥经堆肥后可作为肥料施用于农田,然而关于污泥堆肥过程中抗性基因丰度变化的研究却很少。对污泥堆肥过程中四环素类抗生素(四环素、土霉素和金霉素)和四环素类抗性基因(tet(A), tet(C), tet(M), tet(O)和tet(X))的动态变化进行了定量研究。结果表明,四环素、土霉素和金霉素的浓度经堆肥处理后分别减少85.6%, 91.4% 和 85.3%。高温是导致四环素类抗生素降解的主要因素。经堆肥处理后,tet(A)和tet(X)的相对丰度增加,tet(C)、tet(M)和tet(O)的相对丰度略微下降,说明堆肥处理对污泥中四环素类抗性基因的削减效果有限。     

Effects of soil properties on food web accumulation of heavy metals to the wood mouse (Apodemus sylvaticus)

Effects of soil properties on the accumulation of metals to wood mice (Apodemus sylvaticus) were evaluated at two sites with different pH and organic matter content of the soil. pH and organic matter content significantly affected accumulation of Cd, Cu, Pb and Zn in earthworms and vegetation. For Cd, Cu and Zn these effects propagated through the food web to the wood mouse. Soil-to-kidney ratios differed between sites: Cd: 0.15 versus 3.52, Cu: 0.37 versus 1.30 and Zn: 0.33-0.83. This was confirmed in model calculations for Cd and Zn. Results indicate that total soil concentrations may be unsuitable indicators for risks that metals pose to wildlife. Furthermore, environmental managers may, unintentionally, change soil properties while taking specific environmental measures. In this way they may affect risks of metals to wildlife, even without changes in total soil concentrations.     

鸡粪有机肥对土壤中抗生素抗性基因和整合酶基因的影响

为探究施用有机肥对土壤中抗性基因和整合酶基因分布的影响及其与土壤环境因子的相关关系,利用高通量PCR技术监测了鸡粪有机肥施用120 d后土壤中四环素类抗性基因、大环内酯类抗性基因、整合酶基因和土壤理化性质的变化情况。研究结果表明,施肥土壤的电导率、pH、有机质含量均明显增加,而土壤氧化还原电位由217.27 mV降低到154.47 mV。施肥土壤中钾、氮、磷、铅、铜和锌含量均上升。施加鸡粪有机肥120 d后,土壤中tetM、tetQ、tetW、tetG和tetX 5种四环素类抗性基因相对丰度分别增加了2.90、0.97、6.80、0.98和0.94倍,而erm35、ermB、ermT、ermX和ermF 5种大环内酯类抗性基因相对丰度分别增加了0.98、136.68、0.95、2.89和2.89倍;但erm36相对丰度降低了0.75倍,其中施肥后土壤中ermB丰度最高而erm36丰度最低。施用鸡粪有机肥后,土壤中intI1和intI3的丰度分别降低了4.71 × 10-5和2.57 × 10-7,而intI2的丰度增加了3.74 × 10-6。Network分析发现intI3与除ermB外的基因均呈显著负相关关系(R=-1.00,P<0.05)。     

铅锌冶炼厂周边重金属的空间分布及生态风险评价

以关中西部某铅锌厂周边农田为对象,研究了厂区周边土壤中7种重金属含量的水平及垂直分布特征,用Lars Hakanson潜在生态危害指数对重金属的危害程度进行了评价.研究表明,厂区周围土壤中重金属在水平分布上具有局部高度富集的特征.以厂区为中心,重金属水平分布主要在西北-东南方向上,地势低的土壤中重金属含量明显较高;重金属在垂直分布上主要富集在0～ 25 cm的表层土壤中,且随着深度的进一步增加,土壤中重金属浓度变化幅度减弱,基本趋于稳定.厂区周边Cd和Hg污染水平为重度污染,Pb污染水平为中度污染,潜在生态危害程度为重度.     

某电子垃圾拆解园周边农田土壤中多环芳烃的污染特征及风险评估

研究了某电子垃圾拆解园周边151个农田土壤样品中16种多环芳烃(PAHs)的污染特征和环境风险。结果表明,125个表层土壤样品中PAHs总质量浓度在149.0~2.0×104μg/kg,均值为1 805.5μg/kg,随着剖面土壤深度增加,PAHs含量总体呈递减趋势。通过来源解析,电子拆解园周围土壤中PAHs污染主要由废弃的电子电器元件的粗放燃烧和汽车尾气排放共同引起。土壤风险评估表明,7种类二噁英毒性PAHs的毒性当量(TEQPAH)在6.000×10-5~0.689pg TEQ/g,平均值为0.015pg TEQ/g;苯并(a)芘、二苯并(a,h)蒽、苯并(a)蒽、苯并(b)荧蒽、茚并(1,2,3-cd)芘致癌风险率超出百万分之一的样本比例分别为20.53%、6.62%、1.99%、2.65%、2.65%,其中采样点1、68两个点位表层土壤的苯并(b)荧蒽致癌风险率超过了万分之一。     

Comparing the sensitivity of soil invertebrates to pesticides with that of Eisenia fetida

The sole routine testing of the standard earthworm Eisenia fetida for the terrestrial risk assessment of pesticides has been under much debate since other soil invertebrates may be more sensitive than this standard test species. However, the very low availability of laboratory toxicity data for taxa other than E. fetida has greatly hampered sensitivity comparisons. In the present study, the relative tolerance (T_rel) approach was used to enable comparing toxicity thresholds obtained from the US-EPA ECOTOX database, for main terrestrial taxonomic groups and pesticidal types of action (insecticides, fungicides, herbicides, and other) separately. Analyses confirmed previously reported lower and higher sensitivity of collembolans to fungicides and insecticides, respectively. However, various other discrepancies in susceptibility relative to E. fetida were encountered as indicated by species sensitivity distributions and/or calculated 95% confidence intervals of T_rel values. Arachnids and isopods were found to be more sensitive to insecticides, and nematodes to fungicides, as compared to E. fetida. Implications of study findings for the terrestrial risk assessment of pesticides are discussed.     

Change in the antimicrobial resistance profile of Pseudomonas aeruginosa from soil after exposure to herbicides

The extensive use of pesticides represents a risk to human health and to the environment. This study aimed to investigate if the exposure to atrazine and diuron, two herbicides widely used in Brazil, could induce changes in the susceptibility profile to aztreonam, colistin and polymyxin B antimicrobials in isolates of P. aeruginosa obtained from soil samples by using the determination of minimum inhibitory concentration (MIC) test. Three isolates had an increase of MIC to aztreonam after exposure to both herbicides and one isolate did not show any MIC change. The MexAB-OprM efflux pump has already been upregulated in these isolates and the herbicides atrazine and diuron did not increase MexAB-OprM overexpression. Therefore, the decrease in aztreonam susceptibility was not directly related to this pump, suggesting that probably other mechanisms should be involved.     

Determination of the residue dynamics and dietary risk of thiamethoxam and its metabolite clothianidin in citrus and soil by LC-MS/MS

A modified quick, easy, cheap, effective, rugged and safe (QuEChERS) method was developed for the determination of thiamethoxam and its metabolite clothianidin in citrus (including the whole citrus, peel and pulp) and soil samples by liquid chromatography-tandem mass spectrometry. The sample was extracted with acetonitrile and purified with octadecylsilane. The detection limits of both compounds were 0.0001–0.0002?mg kg^–1, while the limit of quantification of thiamethoxam was 0.002?mg kg^–1 and the limit of quantitation of metabolites was 0.001?mg kg^–1. The recovery was 70.37%–109.76%, with inter-day relative standard deviations (RSD) (n?=?15) values ≤9.46% for the two compounds in the four matrices. The degradation curve of thiamethoxam in whole citrus and soil was plotted using the first-order kinetic model. The half-life of the whole citrus was 1.9–6.2?days, and the half-life of the soil was 3.9–4.2?days. The terminal residue of thiamethoxam (the sum of thiamethoxam and clothianidin, expressed as thiamethoxam) was found to be concentrated on the peel. The final residual amount of thiamethoxam in the edible portion (pulp) was less than 0.061?mg kg^–1. The risk quotient values were all below 1, indicating that thiamethoxam as a citrus insecticide does not pose a health risk to humans at the recommended dosage.     

我国土壤环境管理名录制度实施中的问题分析和对策

我国污染地块土壤环境管理的基本制度包括疑似污染地块名单制度、污染地块名录制度、建设用地土壤污染风险管控和修复名录制度。分析了3项制度的主要内容和执行现状,提出了实施中存在的6个问题,阐述了6个方面的对策建议。分析认为,目前疑似污染地块名单和污染地块名录制度未明确“退出”程序和要求;制度的实际执行情况与管理办法的要求不一致;各地纳入疑似污染地块名单的地块数量明显偏少;信息公开的渠道与内容未做统一要求;初步调查报告的编制和评审要求在现行技术规范中缺乏足够阐述;风险管控和修复名录制度对工程实施的变更程序、是否明确设置工程设计环节等未做出明确规定。最后,提出在污染地块环境管理办法修订过程中应增加“退出”疑似污染地块名单和污染地块名录的条件和要求、信息公开渠道与内容,明确由项目业主单位自行确定是否开展工程设计,将疑似污染地块名录的制定和信息公开责任调升为市级生态环境主管部门负责,并细化《建设用地土壤环境调查评估技术指南》中的初步调查布点要求和采样深度要求,加快省级风险评估报告和效果评估报告评审细则文件的制定等建议,以期为我国土壤环境管理部门提供决策参考。     

Abundance and removal of antibiotic resistance genes (ARGs) in the rearing environments of intensive shrimp aquaculture in South China

Although research regarding antibiotic resistance genes (ARGs) in aquaculture environments has gained increasing scientific interest, further studies are required to understand the abundances and removal mechanisms of ARGs during the entire rearing period of shrimp aquaculture. Thus, in this study, abundances, distributions and removal rates of ARGs in different environmental compartments of intensive shrimp farms in South China were investigated during the entire rearing period. The results indicated that sul1 and cmlA were the predominant ARGs in the water and sediment samples. Additionally, the total abundance of ARGs was higher in shrimp pond water than in the source water and farm effluent. Moreover, sediment samples indicated significantly higher ARG abundances than water samples from the shrimp ponds (P?<?0.05). Environmental factors were found to significantly affect the distribution of ARGs in shrimp rearing environments. Furthermore, stable ponds aided the removal of ARGs from shrimp pond water. This study accounted for temporal variations in ARG abundances as well as removal of ARGs in different environmental compartments during the entire shrimp rearing period. However, additional research is required to optimize the water treatment process for removal of ARGs from the aquaculture.     

复配钝化剂对稻田重金属有效性及其水稻吸收的影响

针对红壤稻米中重金属Cd超标的问题,通过筛选钝化剂并将其复配用于大田实验,以研究复配钝化剂对土壤有效态重金属(Cd、Pb和Cu)及水稻吸收重金属的影响及其作用机理。室内筛选实验表明,在11种无机和有机钝化剂中,海泡石、生石灰和聚丙烯酰胺对土壤重金属钝化效果较好,使有效态Cd、Pb和Cu分别降低了32.4% ~ 89.2%、19.5% ~ 99.1%和49.4% ~ 92.4%,并将它们确定为复配钝化剂的成分。大田实验结果表明,生石灰、聚丙烯酰胺和海泡石复配对土壤中重金属钝化效果最佳,使有效态Cd、Pb和Cu分别降低了28.6%、20.2%和23.5%(p<0.05),其钝化机制为离子交换和络合作用。而且,该复配钝化剂对土壤化学性质影响最小。复配钝化剂使稻米中Cd的质量分数降低了21.7% ~ 93.1%(p<0.05)。另外,复配钝化剂对土壤微生物丰度和多样性没有显著影响。考虑到土壤的安全性和稳定性,推荐将生石灰、聚丙烯酰胺和海泡石复配钝化剂用于降低红壤稻米对Cd的吸收以确保粮食安全生产。本研究结果可为重金属污染红壤稻田的安全利用提供参考。     

The profile of antibiotics resistance and integrons of extended-spectrum beta-lactamase producing thermotolerant coliforms isolated from the Yangtze River basin in Chongqing

The spreading of extended-spectrum β-lactamases (ESBL)-producing thermotolerant coliforms (TC) in the water environment is a threat to human health but little is known about ESBL-producing TCs in the Yangtze River. We received 319 ESBL-producing stains obtained from the Chongqing basin and we investigated antibiotic susceptibility, bla gene types and the presence of integrons and gene cassettes. 16.8% of TC isolates were ESBL-producing bacteria and bla_TEM+CTx-M was the predominant ESBL type. 65.2% of isolates contained class 1 integrons, but only 3 carried intI 2. Gene cassettes were amplified and sequenced. aadA, drfA, cmlA, sat1, aar3 and two ORF cassettes were found. In conclusion, Yangtze River is heavily polluted by ESBL-producing TC bacteria and the combined bla gene type could enhance antibiotic resistance. Class 1 integrons were widespread in ESBL-producing isolates and play an important role in multi-drug resistance. Characterization of gene cassettes could reveal the dissemination of antibiotic resistance genes.     

微量磺胺甲恶唑对饮用水管网生物膜群落及抗性基因的影响与控制

针对饮用水管网系统可能存在的微生物风险问题,采用模拟不同处理条件下的输配水管道系统,通过宏基因组学分析探究微量磺胺甲恶唑以及次氯酸钠消毒对管道中生物膜与抗性基因组成的影响。结果表明,2 μg·L⁻¹磺胺甲恶唑的添加对微生物群落以及抗性基因组成无明显影响,而浮霉菌门细菌表现出很强的抗次氯酸钠消毒能力。在未消毒条件下丰度前十的抗性基因与携带差异性抗性基因的细菌在消毒后丰度均明显有所下降,次氯酸钠消毒使ARGs总量下降了91.9%,因此,次氯酸钠消毒通过控制携带抗性基因物种从而有效控制群落抗性基因的传播。同时,通过组间显著性差异的功能基因与组间显著性差异的抗性基因相关性分析,功能基因的变化情况与抗性基因变化情况一致,因此,长期消毒改变了细菌群落组成及其功能,并最终影响抗性基因传播。这项研究有助于控制长期运行的饮用水管网输配系统中可能存在的包括抗药基因在内的微生物相关风险问题。     

Distribution, availability, and sources of trace metals in different particle size fractions of urban soils in Hong Kong: Implications for assessing the risk to human health

The concentration and loading distribution of trace metals (Cu, Zn, Pb, Co, Ni, Cr, and Mn) and major elements (Al, Ca, Fe, and Mg) in different particle size fractions (2000-280, 280-100, 100-50, 50-10, 10-2, and <2 μm) of surface soils from highly urbanized areas in Hong Kong were studied. The enrichment of Pb, Cu, and Zn in the urban soils was strongly influenced by anthropogenic activities, and Pb accumulated in fine particles was mainly derived from past vehicular emissions as shown by Pb isotopic signatures. Trace metals primarily accumulated in clay, fine silt, and very fine sand fractions, and might pose potential health risks via the inhalation of resuspended soil particles in the air (PM₁₀ or PM_2.5), and ingestion of adhered soils through the hand-to-mouth pathway. The mobility, bioavailability, and human bioaccessibility of Pb and Zn in bulk soils correlated significantly with metal concentrations in fine silt and/or very fine sand fractions.     

The effect of alkyl chain length on the degradation of alkylimidazolium- and pyridinium-type ionic liquids in a Fenton-like system

Background, aim, and scope  Ionic liquids are regarded as essentially “green” chemicals because of their insignificant vapor pressure and, hence, are a good alternative to the emissions of toxic conventional volatile solvents. Not only because of their attractive industrial applications, but also due to their very high stability, ionic liquids could soon become persistent contaminants of technological wastewaters and, moreover, break through into natural waters following classical treatment systems. The removal of harmful organic pollutants has forced the development of new methodologies known as advanced oxidation processes (AOPs). Among them, the Fenton and Fenton-like reactions are usually modified by the use of a higher hydrogen peroxide concentration and through different catalysts. The aim of this study was to assess the effect of hydrogen peroxide concentration on degradation rates in a Fenton-like system of alkylimidazolium ionic liquids with alkyl chains of varying length and 3-methyl-N-butylpyridinium chloride. Materials and methods  The ionic liquids were oxidized in dilute aqueous solution in the presence of two different concentrations of hydrogen peroxide. All reactions were performed in the dark to prevent photoreduction of Fe(III). The concentrations of ionic liquids during the process were monitored with high-performance liquid chromatography. Preliminary degradation pathways were studied with the aid of ¹H NMR. Results  Degradation of ionic liquids in this system was quite effective. Increasing the H₂O₂ concentration from 100 to 400 mM improved ionic liquid degradation from 57–84% to 87–100% after 60 min reaction time. Resistance to degradation was weaker, the shorter the alkyl chain. Discussion  The compound omimCl was more resistant to oxidation then other compounds, which suggests that the oxidation rates of imidazolium ionic liquids by OH· are structure-dependent and are correlated with the n-alkyl chain length substituted at the N-1-position. The level of degradation was dependent on the type of head group. Replacing the imidazolium head group with pyridinium increased resistance to degradation. Nonetheless, lengthening the alkyl chain from four to eight carbons lowered the rate of ionic liquid degradation to a greater extent than changing the head group from imidazolium to pyridinium. 1H-NMR spectra show, in the first stage of degradation, that it is likely that radical attack is nonspecific, with any one of the carbon atoms in the ring and the n-alkyl chain being susceptible to attack. Conclusions  The proposed method has proven to be an efficient and reliable method for the degradation of imidazolium ionic liquids by a Fenton-like reagent deteriorated with lengthening n-alkyl substituents and by replacing the imidazolium head group with pyridinium. The enhanced resistance of 1-butyl-3-methylpyridinium chloride when the resistance of imidazolium ionic liquids decreases with increasing H₂O₂ concentration is probably indicative of a change in the degradation mechanism in a vigorous Fenton-like system. H-NMR spectra showed, in the first stage of degradation, that radical attack is nonspecific, with any one of the carbon atoms in the ring and the n-alkyl chain being susceptible to attack. Recommendations and perspectives  Since ionic liquids are now one of the most promising alternative chemicals of the future, the degradation and waste management studies should be integrated into a general development research of these chemicals. In the case of imidazolium and pyridinium ionic liquids that are known to be resistant to bio- or thermal degradation, studies in the field of AOPs should assist the future structural design as well as tailor the technological process of these chemicals