Bacterial community structure analyses to assess pollution of water and sediments in the Lake Shkodra/Skadar, Balkan Peninsula

- Goal, Scope, Background. Lake Skadar is the largest lake in Balkan Peninsula, located on the Montenegro-Albanian border. The unique features of the lake and wide range of endemic and rare or endangered plant and animal species resulted in the classification of the Skadar as a wetland site of international significance. In spite of its importance the Lake is influenced by inflowing waters from river Morača and other regional rivers contaminated by the industry, municipal and agricultural activities in the area. Therefore, the Lake has been subject of various physical, chemical, biological and toxicological examinations. However, community-level analyses are most relevant to assess the effect of stressors on aquatic ecosystems. In the present study bacterial community structure among differently polluted sites of the lake was compared by genetic fingerprinting technique. Methods Water and sediment samples were collected from five differently polluted sampling sites on the Lake Skadar in spring and autumn of the same year. The bacterial community structure in the samples was characterized and compared by temporal temperature gel electrophoresis (TTGE) analysis of polymerase chain reaction-amplified bacterial 16S rRNA genes. Results and Discussion The TTGE analysis resulted in many distinguishable and reproducible band patterns, allowing reliable comparison of bacterial communities among sampling sites. Results on the bacterial community structure revealed that three of the selected locations can be considered as sites that have not shown any pollution degradation determined by our method, due to similar structure of bacterial community in the sediment samples. On the other hand, significant shifts in bacterial community structure in the mouth of the river Morača and Plavnica were shown. Since the results coincide with some of the bioassays and chemical analysis performed previously, the changes in bacterial community structure are explained as an effect of antropogenic pollution on the lake ecosystem by waters of river Morača and stream Plavnica. Conclusion The TTGE has proven to be an efficient and reliable method to monitor bacterial dynamics and community shifts in aquatic environment, especially in the sediments. Within the variety of environmental quality assessments the use of TTGE analyses of bacterial community is strongly recommended, particularly as an initial investigation. However, in any conclusion on the state of the environment, the TTGE results should be combined to some other biological, chemical and hydrological data. Recommendation and Outlook Since prokaryotes are a crucial group of organisms in the biosphere, the ecosystem function studies are largely based on bacterial communities. Therefore, bacterial community structure analysis should be a part of an integrated weight of evidence approach in pollution assessment. In case of Triad approach, consisting of chemical analyses, bioassays, and community studies in the field, the TTGE bacterial community structure analyses should be placed in the later Triad leg. In comparison to other community studies, based on various biotic indices, the TTGE bacterial community analysis has proven to be very sensitive, reliable and less time consuming.     

环境污染物的分子毒理机制研究进展

以分子生物学技术为基础研究污染物的毒理机制．从分子水平揭示了污染物毒作用的本质。综述了国内外分子毒理机制的研究进展。详细阐述了毒理芯片、单细胞凝胶电泳等分子生物学技术应用与毒理学研究中的特点、应用前景以及存在的问题。提出了分子毒理机制今后的研究方向。     

氯化汞对小鼠睾丸生殖细胞的DNA损伤作用

探索氯化汞对雄性小鼠生殖细胞毒性的分子遗传机制。分别以 0 .0 1、0 .10、1.0 0 mmol/L的氯化汞体外处理小鼠睾丸细胞和以 0 .5、1.0、5 .0 μmol/kg的氯化汞体内暴露小鼠 ,应用单细胞凝胶电泳技术检测生殖细胞 DNA的损伤。体外处理 3种剂量组小鼠睾丸生殖细胞 DNA损伤率显著高于阴性对照组 ( 0 mmol/L组 ,P<0 .0 0 1) ;0 .1、1.0 mmol/L剂量组慧星细胞迁移率显著高于阴性对照组 ( P<0 .0 0 1,P<0 .0 5 )。体内暴露 3种浓度氯化汞组小鼠睾丸生殖细胞 DNA损伤率显著高于阴性对照组 ( 0μmol/kg,P<0 .0 0 1) ,5 .0 μmol/kg组慧星迁移率显著高于阴性对照组 ( P<0 .0 5 )。一定剂量的氯化汞处理引起生殖细胞 DNA损伤作用可能是氯化汞细胞毒性的机制之一。     

Toxicological characterization of the landfill leachate prior/after chemical and electrochemical treatment: A study on human and plant cells

In this research, toxicological safety of two newly developed methods for the treatment of landfill leachate from the Piškornica (Croatia) sanitary landfill was investigated. Chemical treatment procedure combined chemical precipitation with CaO followed by coagulation with ferric chloride and final adsorption by clinoptilolite. Electrochemical treatment approach included pretreatment with ozone followed by electrooxidation/electrocoagulation and final polishing by microwave irradiation. Cell viability of untreated/treated landfill leachate was examined using fluorescence microscopy. Cytotoxic effect of the original leachate was obtained for both exposure periods (4 and 24 h) while treated samples showed no cytotoxic effect even after prolonged exposure time. The potential DNA damage of the untreated/treated landfill leachate was evaluated by the comet assay and cytokinesis-block micronucleus (CBMN) assay using either human or plant cells. The original leachate exhibited significantly higher comet assay parameters compared to negative control after 24 h exposure. On the contrary, there was no significant difference between negative control and chemically/electrochemically treated leachate for any of the parameters tested. There was also no significant increase in either CBMN assay parameter compared to the negative control following the exposure of the lymphocytes to the chemically or electrochemically treated landfill leachate for both exposure periods while the original sample showed significantly higher number of micronuclei, nucleoplasmic bridges and nuclear buds for both exposure times. Results suggest that both methods are suitable for the treatment of such complex waste effluent due to high removal efficiency of all measured parameters and toxicological safety of the treated effluent.     

Bacterial diversity in paclobutrazol applied agricultural soils

The aim of this study was to investigate the bacterial communities on paclobutrazol [(2RS, 3RS)-1-(4-Chlorophenyl)-4, 4-dimethyl-2-(1H-1,2,4-triazol-1-yl) pentan-3-ol]–applied agricultural soils by denaturing gradient gel electrophoresis (DGGE) of polymerase chain reaction (PCR) amplified 16S rDNA gene fragments. Three different agricultural soil samples were collected from paclobutrazol applied mango and waxapple orchards, peanut fields and untreated rice fields as a control for DGGE analysis. The DGGE pattern of PCR- generated 16S rDNA gene fragments indicated that the bacterial populations from four paclobutrazol–applied soils of peanut fields were closely related to each other and two paclobutrazol–applied soils of mango and waxapple orchards harbored closely related bacterial communities. But, paclobutrazol–free agricultural soils comprised relatively a different bacterial group. However, the bacterial populations of mango and waxapple orchard are completely different from the bacterial communities of peanut field. Further purification and sequence analysis of 40 DGGE bands followed by phylogenetic tree assay showed similar results that soil bacteria from paclobutrazol applied mango and waxapple orchard are phylogenetically related. Based on the phylogenetic analysis, the clone M-4 was clad 100 % (bootstrap value) with Mycobacterium sp. The Mycobacterium sp. has been proved to degrade the phenolic compounds such as phenol, 4-chlorphenol, 2,4-dichlorophenol and paclobutrazol molecule containing chlorobenzene ring.     

An impingement plate method to detect deposits of pyrethroid insecticides

Abstract

A silica gel impingement plate for monitoring pyrethroid deposits in environmentally sensitive areas is described. The plate is simple, commercially available, and inexpensive. A residue analysis method is given for deltamethrin deposits with 83.3% mean recovery and a minimum quantifiable limit of 0.005 μg/plate. Pyrethroid deposits are strongly adsorbed to the silica gel, which prevents wash‐off by rainfall and slows photodegradation. In two field experiments, deltamethrin was readily detected 3 weeks after direct spray applications at 6 and 10 g ai/ha. Deltamethrin dissipation on the plates was first‐order with a half‐life of 2.9 ‐ 3.7 d. Photoisomers of deltamethrin were also detected on the plates and the ratio of photoisomers to deltamethrin increased over time. This ratio will indicate the age of deposits in monitoring situations.     

Mapping of tritium emissions using absorption vapour samplers

Püspökszilágy Radioactive Waste Treatment and Disposal Facility (RWTDF) is a typical near-surface engineered repository designated to store low- and intermediate-level wastes from various institutes, research facilities and hospitals in Hungary. Two automatic combined ¹⁴C–tritium sampling units installed at the facility sample the air 2 m above surface. The one installed near the vaults detects tritium (T) activities two orders of magnitude higher than the far reference sampling unit. To localize the T emissions, 19 small absorption vapour samplers filled with silica gel were settled onto the ground surface. After the saturation of the silica gel, the water was recovered and its T concentration was measured with a low-background liquid scintillation counter. The absorption vapour samplers are cheap, simple and easy-to-use. We present the samplers and the T distribution map constructed from the data, which helps to localize the T emission.     

PS缓释处理岩溶水石油污染过程中电子受体的利用

原位化学氧化与生物修复技术在石油污染地下水中具有联合的潜力,但化学氧化对微生物活性具有抑制影响,通过缓释氧化剂技术可能会缓解这种影响。为了更好地认识基于过硫酸钠的缓释技术在修复岩溶地下水石油污染过程中对生物降解作用的影响,本研究利用微元体实验在微氧/厌氧条件下,开展了过硫酸盐缓释处理石油污染修复中不同电子受体利用的研究。实验设置了未投加缓释体和添加缓释体(过硫酸钠/石蜡质量比分别为0.5、1、2和3)的不同工况,通过未添加电子受体、添加100 mg·L⁻¹硝酸盐和添加100 mg·L⁻¹三价铁作为电子受体,探究石油污染物苯、甲苯、二甲苯(简称BTX)以及乙醇(EtOH)处理及电子受体的可利用性。结果表明：BTX的降解速率与缓释体过硫酸钠/石蜡质量比呈正相关,在过硫酸钠/石蜡质量比为2和3时降解速率较高,比乙醇更容易被化学氧化;BTX降解以化学氧化为主,而乙醇的降解则以微生物作用为主。过硫酸盐缓释条件下,生物作用中硝酸盐还原作用较为明显,硫酸盐还原作用较为微弱,铁还原作用不明显;硝酸盐最容易被利用为电子受体,其次为硫酸盐,三价铁被微生物利用的可能性最小。缓释体质量比不同、电子受体类型不同可导致优势菌属存在明显的差异,鞘氨醇单胞菌属和罗尔斯通氏菌属相对丰度普遍较高,二者均可利用多种电子受体降解污染物,在生物降解上起到主要作用。     

The Role of Organic Matter in Natural Aquatic Systems: Analytical Constraints

The dissolved organic matter in aquatic systems consists of a heterogeneous mixture of compounds with wide ranging chemical properties and diverse origins. the presence of organic matter (OM) in surface and ground waters has important implications with regard to the mobility and therefore fate of many potential organic and inorganic pollutants. the majority of the analytical approaches in current use require exhaustive pre-treatment steps which may lead to the formation of artefacts within the sample. an analytical technique based upon gel permeation chromatography is proposed as a more reliable procedure for the characterisation of naturally occurring dissolved organic matter.