Abschätzung der Eignung einer molekularbiologischen Methode zur Charakterisierung der Grundwasserbiozönose

Assessing the Suitability of a Molecularbiological Method To Characterise the Microbial Populations in Groundwater A molecularbiological technique was used to characterise the bacterial community structure of groundwater habitats. This method consists of the isolation of bacterial DNA from the samples, amplification of 16S rDNA by PCR (polymerase chain reaction), and separation of the amplified DNA by DGGE (denaturing gradient gel electrophoresis). By using more specific primer combinations in the PCR instead of universal eubacterial primers, also groups of microorganisms (Proteobacteria, sulfate reducer, Archaea) were determined. The resulting DGGE patterns that reflect the microbial diversity are compared and differences or similarities evaluated. In the present studies, groundwater from different sites (bank filtrate, artificially recharged groundwater, and natural groundwater) and with changing redox milieus (aerobic, anaerobic) were investigated as well as the solid aquifer material. Besides, samples were taken from the different stages of artificial groundwater recharge, i.e., from surface water to the drain tile. Samples from groundwater derived from sites with different hydrogeochemical or hydrological conditions like bank filtrate and recharged groundwater revealed great differences in DGGE patterns indicating a characteristic species composition in these habitats, while samples taken at different times from the same groundwater showed only small seasonal variations. Clearly different patterns were also found for groundwater and the adjacent solid material as well as for anaerobic and aerobic groundwaters. Looking at artificial groundwater recharge, almost identical patterns were found in raw water and samples from gravel and sand filtration. DGGE patterns from the resulting groundwater indicated a total change in community structure during underground passage. By using group specific primers, Desulfovibrionaceae, Desulfobacteriaceae, and Archaea could be detected in anaerobic groundwaters.The molecularbiological approach described here gives an increasingly comprehensive and more precise picture of the microbial population of different environments. It is especially suitable to compare the community structure from different habitats or to analyse changes for example due to environmental stress at the same site.     

Biogeographical distribution and diversity of bacterial and archaeal communities within highly polluted anoxic marine sediments from the marmara sea

Physicochemical and microbiological characterization of anoxic sediments taken from seven highly polluted sites of the Marmara Sea was carried out. The 16S rRNA based microbial community structure analyses were performed using domain-specific PCR followed by denaturant gradient gel electrophoresis (DGGE) and sequencing of characteristic bands. The results showed that the microbial communities in these sediments were diverse and evenly distributed. Relating the prokaryotic and geochemical variables through statistical tools revealed that the microbial diversity in the sediments significantly related to depth, and S, Mn and Fe content of the sediments. Fermentative bacteria, denitrifying bacteria and hydrogenotrophic methanogens were dominant whereas sulfate reducing bacteria were absent in the DGGE patterns. This unusual microbial community structure implied that the newly discovered anaerobic methane oxidation coupled to denitrification process may occur in these subseafloor environments.     

Denaturing gradient gel electrophoresis shows that bacterial communities change with mid-ocean ballast water exchange

Ships carry ballast water for better stability and to control trim. However, the discharge of ballast water near ports is known to transport invasive species from one coastal area to another. The exchange of ballast water on the high seas is supposed to reduce such invasions of exotic species. In this study, we used denaturing gradient gel electrophoresis (DGGE) to analyze the composition of the bacterial community in ballast water before and after such a mid-ocean exchange, and we also measured total bacterial counts. Our findings confirmed that the ballast water was replaced by the mid-ocean exchange, as indicated by the marked change in the composition of the bacterial community. There was also a significant decrease in bacterial abundance after the mid-ocean exchange. Finally, our findings support the incubation hypothesis, because the composition of the bacterial communities changed over time within the same ballast water.     

人工回灌过程中地下水微生物群落变化

人工回灌过程中,回灌水的注入使目的含水层地下水环境发生变化,微生物条件也会随之改变,从而影响地下水环境质量及水文地球化学作用。以上海市某人工回灌试验场为例,在分析人工回灌过程中水化学演化特点的基础上,应用DGGE技术对场地回灌过程中地下水中的微生物群落结构变化进行研究,为评价人工回灌对地下水水质安全的影响提供科学的理论依据。结果表明:人工回灌作用使目的含水层地下水中的Eh值及DO质量浓度升高,分别由64.0 mV、1.12 mg/L升至534.4 mV、1.44 mg/L;同一位置处微生物群落结构与原始地下水状态的相似性随时间降低;同一时刻距离回灌井越远的监测井的微生物群落结构越接近于原始地下水状态。随着回灌的进行,目的含水层地下水中优势菌属(种)共有7种,其中Rubrivivax gelatinosus和Candidatus Accumulibacter phosphatis clade IIA str.的反硝化能力以及Rhodoferax ferrireducens对Fe³⁺的还原能力,对地下水水化学组分产生影响。     

PAHs contamination and bacterial communities in mangrove surface sediments of the Jiulong River Estuary, China

Sixteen sediment samples collected from eight transects in a mangrove swamp of the Jiulong River Estuary, Fujian, China were investigated for their content of polycyclic aromatic hydrocarbons (PAHs) and the biodegradation potential of the indigenous microorganisms. The bacterial community structures in the mangrove sediments and in enrichment cultures were also investigated. The results showed that the total PAHs concentration of mangrove sediments ranged from 280 to 1074 ng g(-1) dry weight, that the PAHs composition pattern in the mangrove sediments was dominated by high molecular weight PAH components (4-6 rings), and that Benzo[ghi]perylene and Indeno[1,2,3-cd]pyrene were the most dominant at different stations. Abundant PAH-degrading bacteria were found in all the stations, the values of phenanthrene-degrading bacteria ranged from 5.85 x 10(4) to 7.80 x 10(5) CFU g(-1) dry weight, fluoranthene-degrading bacteria ranged from 5.25 x 10(4) to 5.79 x 10(5) CFU g(-1) dry weight, pyrene-degrading bacteria ranged from 3.10 x 10(4) to 6.97 x 10(5) CFU g(-1) dry weight and the benzo(a)pyrene-degrading bacteria ranged from 5.25 x 10(4) to 7.26 x 10(5) CFU g(-1) dry weight. DGGE analysis of PCR-amplified 16S rDNA gene fragments confirmed that there was a remarkable shift in the composition of the bacterial community due to the addition of the different model PAH compound phenanthrene (three ring PAH), fluoranthene(four ring PAH), pyrene(four ring PAH) and benzo(a)pyrene(five ring PAH) during enrichment batch culture. Eleven strains were obtained with different morphology and different degradation ability. The presence of common bands for microbial species in the cultures and in the native mangrove sediment DNA indicated that these strains could be potential in situ PAH-degraders.     

氨氮污染源区包气带剖面微生物生态分布特征的研究

包气带土壤中氨氮污染迁移规律已成为近年来国内外学者研究的重点,目前在氨氮污染迁移规律研究中采用柱实验模拟以及软件模拟的方法较多,土壤生物技术则广泛应用于污染物降解领域,而在污染物迁移规律研究中应用较少。本文采用变性梯度凝胶电泳(DGGE)技术、16S rRNA序列分析技术以及典范对应分析相结合,对华北平原3个典型氨氮污染区土壤表层至包气带剖面微环境的细菌垂直分布特征及群落结构进行研究。结合污染区土壤理化性质分析,认为包气带土壤剖面的细菌群落中存在着与氮循环、硫酸盐代谢等过程偶联的优势细菌类群,说明土壤微环境中细菌群落分布明显受氨态氮、硝态氮、亚硝态氮和硫酸盐的分布影响,进一步表明污染土壤优势菌群的群落结构信息是描述包气带土壤环境氨氮污染物迁移规律的重要参数。     

Metabolic and Phylogenetic Profile of Bacterial Community in Guishan Coastal Water（Pearl River Estuary）, South China Sea

Characteristics of a microbial community are important as they indicate the status of aquatic ecosystems. In the present study, the metabolic and phylogenetic profile of the bacterioplankton community in Guishan coastal water（Pearl River Estuary）, South China Sea, at 12 sites（S1–S12） were explored by community-level physiological profiling（CLPP） with BIOLOG Eco-plate and denaturing gradient gel electrophoresis（DGGE）. Our results showed that the core mariculture area（S6, S7 and S8） and the sites associating with human activity and sewage discharge（S11 and S12） had higher microbial metabolic capability and bacterial community diversity than others（S1–5, S9–10）. Especially, the diversity index of S11 and S12 calculated from both CLPP and DGGE data（H 3.2） was higher than that of others as sewage discharge may increase water nitrogen and phosphorus nutrient. The bacterial community structure of S6, S8, S11 and S12 was greatly influenced by total phosphorous, salinity and total nitrogen. Based on DGGE fingerprinting, proteobacteria, especially γ- and α-proteobacteria, were found dominant at all sites. In conclusion, the aquaculture area and wharf had high microbial metabolic capability. The structure and composition of bacterial community were closely related to the level of phosphorus, salinity and nitrogen.