Variations of Bacterial Community Structure in Flooded Paddy Soil Contaminated with Herbicide Quinclorac

The denaturing gradient gel electrophoresis (DGGE) method was applied to determine the relative genetic complexity of microbial communities in flooded paddy soil treated with herbicide quinclorac (3,7-dichloro-8-quinoline-carboylic acid). The results obtained showed a significant effect of quinclorac on the development of bacterial populations in soils contaminated with different concentrations of the herbicide at the early time after application. In general, however, the number of populations of the same soil sample treated with the same concentration of the quinclorac differed obviously with increasing incubation time within the early 8 weeks. The scale of differences in banding patterns-showed that the microbial community structures of the quinclorac-treated and non-quinclorac-treated soils were not significantly different after 21 weeks of incubation. Quantification, as demonstrated in this paper, was studied by establishing dose-response relationships. Significant pattern variations were quantified. Prominent DGGE bands were excised, cloned and sequenced to gain insight into the identities of predominant bacterial populations. The majority of DGGE band sequences were related to bacterial genera Clostridium, Sphingobacterium, Xanthomonas and Rhodococcus.     

氨氧化菌群的筛选及发酵条件的优化

利用选择性培养基对活性污泥进行连续驯化,筛选出氨氮去除效率较高且稳定的氨氧化菌群。采用聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)技术分析了氨氧化菌群在连续传代过程中菌群的结构差异,并对菌群的发酵培养基及发酵条件分别进行了正交优化和单因子优化。结果表明,该菌群的最佳发酵培养基为每升培养基中含碳酸盐缓冲液15mmol,硫酸铵4.2 mmol,磷酸盐缓冲液12.5 mmol,硫酸亚铁0.9μmol,氯化钙0.4 mmol,硫酸镁1.5 mmol;最佳发酵条件为:接种量为14%,装液量为60 mL/250 mL,温度为35℃。在此条件下,菌群对氨氮的去除效果较优化前提高了155%。     

高效氨氧化菌群富集、驯化及其动态变化规律分析

利用选择性培养基对氨氧化菌群进行了连续驯化,得到了氨氮去除效率稳定的氨氧化菌群。采用平板菌落计数法结合聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)技术考察了氨氧化菌群在连续传代过程中数量及群落结构的动态变化,并考察了pH值、碳源(HCO3-)浓度和氨氮浓度等因素对氨氧化菌群去除氨氮效率的影响。结果表明,通过连续驯化,氨氧化菌的比例由最初的1.8%提高到了31.3%。在碳源浓度为1.5 mg/L,NH4+-N初始浓度为200 mg/L,pH值为8的条件下,菌群对氨氮的去除率达99%以上。     

Influence of tetracycline exposure on the growth of wheat seedlings and the rhizosphere microbial community structure in hydroponic culture

In this study, the effects of tetracycline exposure on wheat growth and the microbial community structure in the rhizosphere were investigated under hydroponic culture conditions. Exposure to various concentrations of tetracycline resulted in significant suppression of the growth of wheat roots and shoots, with minimum doses of 0.8 mg L^?1 and 4 mg L^?1 resulting in inhibition rates of 32% and 15.4%, respectively. Complete inhibition of the growth of these two parts of wheat plants was observed in response to treatment with tetracycline at 20 mg L^?1 and 100 mg L^?1, respectively. However, the germination of wheat seeds was not sensitive to exposure to tetracycline. The effects of tetracycline exposure on the microbial community in the wheat rhizosphere were evaluated through traditional cultivation and molecular biological analyses. The cultivation results indicated that bacteria were the dominant population, being present in concentrations of 1× 10⁸–2.45× 10⁹CFUs mL^?1, although 39% to 87% inhibition occurred in response to tetracycline. The concentration of fungi increased in all tetracycline treated samples to 2.5 to 15.8 times that of the control. The highest concentration of fungi (4.27× 10⁸ CFU mL^?1) was observed in response to 60 mg L^?1 tetracycline after 15 days of cultivation. In this stage, a large amount of fungal colonies was observed on the surface of the culture solution, the wheat roots became rotted and the plants became atrophic or even died. Molecular biological analysis indicated that the bacterial community structure was significantly different in samples that were exposed to high levels of tetracycline (over 20 mg L^?1) than in samples that were exposed to lower concentrations. As the concentration of tetracycline increased, the diversity of the bacteria decreased. Additionally, several dominant sensitive species such as Sphingobacterium multivorum were suppressed by tetracycline, while some resistant species such as Acinetobacter sp. appeared or were conserved. The bacteria population tended to stabilize when the drug concentration exceeded 40 mg L^?1.     

Effects of fungicides triadimefon and propiconazole on soil bacterial communities

The impact of fungicides triadimefon and propiconazole on soil bacterial populations from a strawberry field was investigated. Two fungicides were applied to the soil at concentrations of 10 mg/kg or 100 mg/kg with soil water contents 20.2% (fresh soil water content) or 26.0% (field capacity). Changes in bacterial communities were assessed using DNA extraction, polymerase chain reaction (PCR) amplification of the 16S rDNA and denaturing gradient gel electrophoresis (DGGE). High performance liquid chromatography (HPLC) was utilized to detect the residue of fungicides in soils. The results showed that propiconazole was more persistent than triadimefon in soils, and the two soil water contents did not cause significant differences in dissipation rates between the two fungicides. A high concentration of propiconazole could inhibit the existence of soil microbes while one of triadimefon might induce the microbial population in the first stage. From unweighted pair-group method using arithmetic averages (UPGMA) dendrograms, the effect of triadimefon and propiconazole at the two applied concentrations on a soil bacterial community could be long term. After triadimefon was applied for 60 days and propiconazole for 75 days, the compositions of microbial communities were not recovered. From the viewpoint of environmental protection, it was of significant importance to pay more attention not only to the residues of pesticide but also to the change in soil microbial communities.     

活性污泥总DNA提取方法的比较

从处理某制药废水的MBR反应器中采集活性污泥,评价不同DNA提取方法对其总DNA提取效率的影响。DNA提取分细胞裂解和DNA纯化2步,对细胞裂解比较了珠磨匀浆法、反复冻融法、十二烷基磺酸钠（sodium dodecyl sulfate,SDS）裂解法等7种方法;对DNA纯化比较了酚/氯仿纯化法和胶回收纯化法。结果表明,SDS裂解法、酚氯仿纯化法最优。通过条件优化实验,确定SDS裂解酚/氯仿纯化法在污泥量1.1 g,10 000 r/min离心5 min的操作条件下,获得的DNA产量（10 774 μg/g泥重）和纯度（OD₂₆₀∶OD₂₈₀=1.84）等综合指标最好。     

Metagenomic assessment of the microbial diversity in ground pork products from markets in the North Central Region of South Korea

The purpose of this study was to characterize the microbial community in ground pork using molecular approaches. Forty six ground pork products were purchased from local stores in the north central area of South Korea. Aerobic plate counts varied 4.23 ± 5.14 × 10⁵ CFU/g with the range between 5.00 × 10³ and 1.85 × 10⁶ CFU/g for ground pork samples. Four ground meat samples were further processed for metagenomic analysis. Pseudomonas species was the most relative abundant with a wide range occurring (1.72 to 77.7%) as part of the microbial genera in ground pork. Bacteria such as Carnobacterium, Yersinia, Photobacterium were also identified in ground pork. Despite the prominence of certain genera across all samples there was still extensive microbial diversity among ground pork products that originated from different slaughter houses and were processed in different markets. Such diversity indicates that designing interventions to extend shelf life may be hampered by the extensive variability in the microbial consortia associated with pork products. However, this diversity may be useful for developing microbial traceability signatures unique to a slaughter house or a particular market.     

Effect of Dursban 480 EC (chlorpyrifos) and Talstar 10 EC (bifenthrin) on the physiological and genetic diversity of microorganisms in soil

This investigation was undertaken to determine the impact of the insecticides Dursban 480 EC (with organophosphate compound chlorpyrifos as the active ingredient) and Talstar 10 EC (with pyrethroid bifenthrin as the active ingredient) on the respiration activity and microbial diversity in a sandy loam luvisol soil. The insecticides were applied in two doses: the maximum recommended dose for field application (15 mg kg^?1 for Dursban 480 EC and 6 mg kg^?1 for Talstar 10 EC) and a 100-fold higher dose for extrapolation of their effect. Bacterial and fungal genetic diversity was analysed in soil samples using PCR DGGE and the functional diversity (catabolic potential) was studied using BIOLOG EcoPlates at 1, 3, 7, 14, 28, 56 and 112 days after insecticide application. Five bacterial groups (α, β, γ proteobacteria, firmibacteria and actinomycetes) and five groups of fungi or fungus-like microorganisms (Ascomycota, Basidiomycota, Chytridiomycota, Oomycota and Zygomycota) were analysed using specific primer sets. This approach provides high resolution of the analysis covering majority of microorganisms in the soil. Only the high-dose Dursban 480 EC significantly changed the community of microorganisms. We observed its negative effect on α- and γ-proteobacteria, as the number of OTUs (operational taxonomic units) decreased until the end of incubation. In the β-proteobacteria group, initial increase of OTUs was followed by strong decrease. Diversity in the firmibacteria, actinomycetes and Zygomycota groups was minimally disturbed by the insecticide application. Dursban 480 EC, however, both positively and negatively affected certain species. Among negatively affected species Sphingomonas, Flavobacterium or Penicillium were detected, but Achromobacter, Luteibacter or Aspergillus were supported by applied insecticide. The analysis of BIOLOG plates using AWCD values indicated a significant increase in metabolic potential of microorganisms in the soil after the high-dose Dursban application. Analysis of respiration demonstrated high microbial activity after insecticide treatments; thus, microbial degradation was relatively fast. The half-life of the active insecticide compounds were estimated within the range of 25 to 27 days for Talstar and 6 to 11 days for Dursban and higher doses stimulated degradation. The recommended dose levels of both insecticides can be considered as safe for microbial community in the soil.     

A sensitive monoclonal antibody-based enzyme-linked immunosorbent assay for chlorpyrifos residue determination in Chinese agricultural samples

A monoclonal antibody-based competitive antibody-coated enzyme-linked immunosorbent assay (ELISA) was developed and optimized for determining chlorpyrifos residue in agricultural products. The IC₅₀ and IC₁₀ of this ELISA were 3.3 ng/mL and 0.1 ng/mL respectively. The average recoveries in six agricultural products were between 79.5% and 118.0%, with the intra-assay coefficient of variation being less than 8 %. The limit of detection for all tested products was 30 ng/g. To the best of our knowledge, this assay has the best specificity among all the published research on ELISAs for chlorpyrifos.     

Persistence and distribution of pesticide residues in fresh agricultural food consumed in the province of Bologna

The presence of pesticide residues in fresh vegetables and fruit have been qualitatively and quantitatively determined at the laboratories of the Regional Agency for Environmental Protection (ARPA), Division of the Province of Bologna. More than 1,700 samples have been tested by routine analyses. The possible risks for consumers have been evaluated by various parameters. The most important ones were: the amount of each residue; the respective ADI (Acceptable Daily Intake) limit; the contemporary presence of different residues; an estimation of the daily intake, based on the amount of fruit and vegetables consumed per person. It has been possible to evaluate that the daily intake of pesticide residues in the province of Bologna during the period 2003–06 resulted lower than the ADI limits concerning the vegetables. According to the information on fruit consumption the daily intake of omethoate (O,O-dimethyl S-methylcarbamoylmethyl phosphorothioate) resulted higher than its ADI limit, of dicofol (2,2,2-trichloro-1,1-bis(trichloromethyl)benzenemethanol) very close to the admitted limit, under the respective limits for all the other residues.     

不同前处理方式对铁屑还原降解土壤中对硝基苯酚的影响

为推动铁屑在治理受硝基酚类化合物污染土壤中的实际应用,常温(25±1℃)常压下,利用不同前处理方式处理的铁屑、还原铁粉对土壤中的对硝基苯酚(p-NP)进行了还原降解研究;分析了土壤部分理化性质对p-NP还原效果的影响;并对反应时间、铁屑用量和土壤含水量3个人工易控因素作了最优化选择。结果表明:铁屑和铁粉对p-NP还原降解效果影响的大小顺序为:酸洗铁屑还原铁粉水洗铁屑碱洗铁屑;适中的土壤含水量、偏酸性的土壤初始pH值及较高的土壤有机质含量均可显著提高铁屑对p-NP的还原率;正交实验结果显示反应时间对p-NP还原效果影响最大,铁屑用量次之,土壤含水量最小;处理1.5 g p-NP浓度约为1.3×10-5mol/g的模拟污染土壤的反应最优化条件为:酸洗铁屑用量26 mg,土壤含水量0.35 mL,反应时间130 m in,还原率可达到96.4%。     

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.     

甘蔗渣修复铬污染土壤的效果

通过室内模拟培养实验,探讨了甘蔗渣修复铬污染土壤的效果、土壤中六价铬初始浓度和甘蔗渣添加量对六价铬还原反应一级动力学的影响、微生物对甘蔗渣修复铬污染土壤效果的影响及甘蔗渣修复铬污染土壤的机理。结果表明,甘蔗渣能有效地降低污染土壤中铬的浸出毒性和去除土壤中的六价铬。当甘蔗渣的添加量为5%,六价铬浓度低于1 740 mg·kg-1时,培养70 d内,土壤样品的浸出液中六价铬未检出,培养90 d内,土壤中六价铬的去除率趋近100%。土壤中六价铬的还原反应速率随六价铬初始浓度的增加而减小,随甘蔗渣添加量的增加而增大。同时,灭菌和未灭菌条件下,甘蔗渣对铬污染土壤的修复效果差异性不显著。甘蔗渣修复铬污染土壤的机理可能是甘蔗渣中的蔗糖和纤维素先降解生成葡萄糖和果糖,接着葡萄糖和果糖将土壤中的六价铬还原成三价铬。     

The formation and fate of chlorinated organic substances in temperate and boreal forest soils

Background, aim and scope  Chlorine is an abundant element, commonly occurring in nature either as chloride ions or as chlorinated organic compounds (OCls). Chlorinated organic substances were long considered purely anthropogenic products; however, they are, in addition, a commonly occurring and important part of natural ecosystems. Formation of OCls may affect the degradation of soil organic matter (SOM) and thus the carbon cycle with implications for the ability of forest soils to sequester carbon, whilst the occurrence of potentially toxic OCls in groundwater aquifers is of concern with regard to water quality. It is thus important to understand the biogeochemical cycle of chlorine, both inorganic and organic, to get information about the relevant processes in the forest ecosystem and the effects on these from human activities, including forestry practices. A survey is given of processes in the soil of temperate and boreal forests, predominantly in Europe, including the participation of chlorine, and gaps in knowledge and the need for further work are discussed. Results  Chlorine is present as chloride ion and/or OCls in all compartments of temperate and boreal forest ecosystems. It contributes to the degradation of SOM, thus also affecting carbon sequestration in the forest soil. The most important source of chloride to coastal forest ecosystems is sea salt deposition, and volcanoes and coal burning can also be important sources. Locally, de-icing salt can be an important chloride input near major roads. In addition, anthropogenic sources of OCls are manifold. However, results also indicate the formation of chlorinated organics by microorganisms as an important source, together with natural abiotic formation. In fact, the soil pool of OCls seems to be a result of the balance between chlorination and degradation processes. Ecologically, organochlorines may function as antibiotics, signal substances and energy equivalents, in descending order of significance. Forest management practices can affect the chlorine cycle, although little is at present known about how. Discussion  The present data on the apparently considerable size of the pool of OCls indicate its importance for the functioning of the forest soil system and its stability, but factors controlling their formation, degradation and transport are not clearly understood. It would be useful to estimate the significance and rates of key processes to be able to judge the importance of OCls in SOM and litter degradation. Effects of forest management processes affecting SOM and chloride deposition are likely to affect OCls as well. Further standardisation and harmonisation of sampling and analytical procedures is necessary. Conclusions and perspectives  More work is necessary in order to understand and, if necessary, develop strategies for mitigating the environmental impact of OCls in temperate and boreal forest soils. This includes both intensified research, especially to understand the key processes of formation and degradation of chlorinated compounds, and monitoring of the substances in question in forest ecosystems. It is also important to understand the effect of various forest management techniques on OCls, as management can be used to produce desired effects.     

Impact of neighbourhood land-cover in epiphytic lichen diversity: analysis of multiple factors working at different spatial scales

The objective of this work was to determine the impact of neighbourhood land-cover in epiphytic lichen diversity. We used geostatistics to analyse the spatial structure of lichen-indicators (number of lichen species and Lichen Diversity Value) and correlate them to land-cover considering different distances from the observed data. The results showed that lichen diversity was influenced by different environmental factors that act in the same territory but impact lichens at different distances from the source. The differences in the distance of influence of the several land-cover types seem to be related to the size of pollutants/particles that predominantly are dispersed by each land-cover type. We also showed that a local scale of analysis gives a deeper insight into the understanding of lichen richness and abundance in the region. This work highlighted the importance of a multiple spatial scale of analysis to deeply interpret the relation between lichen diversity and the underling environmental factors.     

福州市蔬菜基地土壤重金属和多环芳烃的含量及其与土壤磁性指标的关系

蔬菜基地土壤污染状况与人类健康有着密切的关系。本研究对福州市蔬菜基地土壤中6种重金属和15种美国环境保护总署（US EPA）优控的多环芳烃（PAHs）污染状况进行了调查,并分析了污染物含量与土壤磁性指标间的关系。结果表明,土壤中重金属平均含量为Zn 147.0 mg·kg-1、Pb 55.0 mg·kg-1、Cu 45.5 mg·kg-1、Cr 21.3 mg·kg-1、As 16.0 mg·kg-1、Cd 0.6 mg·kg-1;PAHs的总量范围为324.3~1 838.6 μg·kg-1。Cu含量和PAHs总量与土壤磁化率χ都具有显著的相关性（pχfd的结果分析表明,土壤中PAHs主要来源于石油泄漏和生物质的燃烧;重金属来源复杂,为混合来源,包括人类活动排放和成土母质固有含量。污染评价结果表明,土壤中Cr和As的含量都未超标,Cu、Zn和Cd的含量均超标;参考荷兰土壤修复标准中的目标值, PAHs含量超标的土壤为11.1%,而参照我国土壤污染状况评价技术规定,PAHs含量超标的土壤为22.2%。     

Pollution characteristics of 23 veterinary antibiotics in livestock manure and manure-amended soils in Jiangsu province,China

The aim of this study was to investigate the pollution characteristics of typical veterinary antibiotics in manure and soil of livestock farms in Jiangsu province. This investigation employed solid-phase extraction (SPE) coupled with ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). A total of 53 manure and 50 amended soil samples from 16 livestock farms in Jiangsu province were collected for analysis. In the manure samples, the highest detected frequencies and concentrations were those of tetracyclines (TCs, 54.1 ± 5775.6 μgkg^?1), followed by fluoroquinolones (FQs, 8.4 ± 435.6 μgkg^?1), sulphonamides (SAs, 3.2 ± 5.2 μgkg^?1) and macrolides (MACs, 0.4 ± 110.5 μgkg^?1). Statistical analysis was used to illuminate the pollution characteristics of 23 veterinary antibiotics for various animal types and different regions in Jiangsu province. The results showed that the pollution level in cow manure was relatively lower compared with pig and chicken manure due to the relative restriction of medication. Furthermore, contamination was serious in amended soil from chicken farms. The pollution level in manure among different regions was higher to the south and north compared with the centre of the region. The same outcome was found for soil. Antibiotic residues in organic fertilizer were also investigated in this study. We found that although the detected concentration was lower in organic fertilizer than in fresh manure, detection frequencies (10–90%) were high, especially for roxithromycin (90%) in MACs (30–90%). This finding suggests attention should be paid to the pollution levels in organic fertilizer. This study is the first extensive investigation of the occurrence and distribution of many kinds of typical veterinary antibiotics in manure and soil from livestock farms of Jiangsu province. This investigation systematically assesses veterinary antibiotics usage and related emissions in southeast China.