RAPD Marker and Substrate Utilization Pattern Applied to Study Microbial Community Diversity in the Soil Affected by Agricultural Chemicals

Abstract

Present analyses of random amplified polymorphic DNA (RAPD) and Biolog GN substrate utilization pattern are combined to further study the diversity of microbial communities in four soils affected by agricultural chemicals. The results showed that the four soil microbial communities were apparently distinguishable in the diversity at RAPD level in terms of the richness and modified richness in the summer, which supports our previous report using the same soils in winter. A significant difference for the average well color development (AWCD) at 72 h incubation was found among the soils in winter using Biolog GN substrate utilization pattern, but this difference was not found among the soils in summer. However, Shannon-Weaver indices for microbial communities in the summer soils polluted by agricultural chemicals were significantly higher than those in winter at metabolic level; in contrast, no significant difference existed between the two seasons for microbial communities in the soil without chemical pollution. Present results suggest that the combined approach using RAPD and substrate utilization pattern could be used to effectively quantify microbial community diversity and its changes among the seasons in the soils affected by agricultural chemicals, simultaneously at molecular and physiological levels.     

Studies on Formation of Liquid and Gaseous Formaldehyde-Induced DNA-Protein Crosslinks in Rat Marrow Cells

甲醛是一种遗传毒物,流行病学研究表明甲醛可能具有诱导白血病的作用,然而甲醛诱导白血病的机制目前还不清楚. 以不同浓度液态和气态甲醛对大鼠骨髓细胞进行染毒,采用KCl-SDS 法检测了骨髓细胞 DNA-蛋白质交联程度,并采用单细胞凝胶电泳技术(彗星实验)检测了骨髓细胞 DNA 链断裂程度. 研究结果表明:与对照组相比,低浓度甲醛(液态甲醛浓度为:5μmol·L-1和25μmol·L-1;气态甲醛浓度为:0.5mg·m-3 和 1.0mg·m-3)可以引起 DNA断裂水平显著增高 (p<0.01);而高浓度甲醛 (液态甲醛浓度为:125μmol·L-1 和 625μmol·L-1;气态甲醛浓度为:3.0mg·m-3)则可以引起 DNA-蛋白质交联水平显著增高(p<0.01; p<0.05). 研究结果提示:甲醛染毒可以导致大鼠骨髓细胞DNA的损伤,暗示甲醛诱导白血病具有高度的可能性.     

Genotoxicity monitoring of freshwater environments using caged crayfish (Astacus leptodactylus)

Genotoxicity of freshwater pollution was assessed by measuring DNA damage in haemocytes of caged freshwater crayfish Astacus leptodactylus by the means of Comet assay and micronucleus test, integrated with the measurements of physiological (total protein concentration) and immunological (total haemocyte count) haemolymph parameters as biomarkers of undergone stress. Crayfish were collected at the reference site (River Mre?nica) and exposed in cages for 1 week at three polluted sites along the Sava River (Zagreb, Sisak, Krapje). The long term pollution status of these locations was confirmed by chemical analyses of sediments. Statistically significant increase in DNA damage measured by the Comet assay was observed at all three polluted sites comparing to the crayfish from reference site. In addition, native crayfish from the mildly polluted site (Krapje) cage-exposed on another polluted site (Zagreb) showed lower DNA damage than crayfish from the reference site exposed at the same location indicating adaptation and acclimatisation of crayfish to lower levels of pollution. Micronuclei induction showed similar gradient of DNA damage as Comet assay, but did not reach the statistical significance. Observed increase in total haemocyte count and total protein content in crayfish from polluted environments in the Sava River also confirmed stress caused by exposure to pollution. The results of this study have proved the applicability of caging exposure of freshwater crayfish A. leptodactylus in environmental genotoxicity monitoring using Comet assay and micronucleus test.     

水中细菌内毒素污染特性及检测方法研究进展

细菌内毒素是革兰氏阴性菌和某些蓝藻细胞壁的脂多糖复合物,主要由菌体死亡解体后释放,是常见的外源性致热原,具有强免疫刺激能力,与人类多种疾病密切相关.内毒素分子的耐受力好,耐热性强,不易被常规方法去除.饮用水和水环境中的内毒素可通过多种暴露途径导致机体潜在的健康风险,成为近年来研究的热点.本文概述了细菌内毒素的理化特性和生物活性,综述了细菌内毒素多种检测方法的研究进展,并对细菌内毒素的法定检测方法——鲎试验用于检测水体环境样品的干扰问题和排除方法进行分析.     

15N Metabolic test for the determination of phytotoxic effects of chemicals and contaminated environmental samples

A stable isotope¹⁵N-nitrogen test (ESIMA = Ecotoxicological Stable Isotope Metabolic Assay) was developed to assess biological effects and the potential toxicological hazard of chemicals and contaminated environmental samples on plant metabolism. The assay measures the effect of toxicants on the incorporation of a¹⁵N labelled tracer into the total nitrogen fraction (both the nonprotein and protein fraction) of plants. Segments ofPisum arvense epicotyls are used as test substrates because of their high metabolic activity. The plant material is incubated under standardised conditions for two hours; subsequently¹⁵N incorporation is analysed by determining the¹⁵N abundance (¹⁵N atom-%) in the epicotyl segments. The effects of toxicants are evaluated by comparing the¹⁵N incorporation rates of control tissue and epicotyl segments exposed to individual chemicals or complex environmental samples. The specificity and sensitivity of effects as indicated by ESIMA were compared with effects as measured by two established ecotoxicological bioassays, the pollen tube growth test using pollen ofNicotiana sylvestris and the bacterial luminescence inhibition test using pollen ofPhotobacterium phosphoreum. The results of the study clearly indicate the suitability of ESIMA for assessing toxic impacts on plant nitrogen metabolism. Prof. Dr. habil. Hans Faust dedicated to his 70^th birthday.     

一株生物素依赖型菌株bio-5:分离,初步鉴定及其用于生物素浓度测定

从西红柿根圈土壤中分离纯化到一株生物素依赖型菌株bio - 5 ,经测定其内源抗生素抗性和脂肪酸特征 ,初步定名为假单胞菌bio - 5 .这一菌株对生物素非常敏感 .建立了以此菌株用于测定生物素浓度的标准方法 .利用这一方法 ,测定了不同作物叶子中生物素的含量 ,并将这一方法成功用于筛选转stv基因烟草植物 .与传统生物素测定菌株LactobacillusplantarumATCC80 14比较 ,我们所分离的菌株具有易于获得 ,易于培养 ,快速用于生物素浓度测定的特点 .图 7表 2参 2 2     

植物病原细菌的BIOLOG系统鉴定及其多元统计初析

Ｂｉｏｌｏｇ细菌鉴定系统已广泛应用于细菌快速鉴定 ,通过比较菌株碳源利用的代谢指纹 ,还可用于细菌分类研究[1～ 4 \〗.该系统具有自动化和标准化程度高、结果快速准确和操作简便等优点 ,它主要根据细菌对 95种碳源的利用情况 ,以四唑紫 (Ｔｅｔｒａｚｏｌｉｕｍ)为指示剂 ,Ｂｉｏｌｏｇ代谢指纹经计算机处理并与数据库内已知细菌比较 ,实现对待测菌的快速鉴定 .Ｂｉｏｌｏｇ系统用于细菌分类研究中 ,除已广泛采用聚类分析并以树状图表示菌株间的相似关系外 ,近年来 ,国外许多学者已应用多元统计方法 (主成分分析、因子分析和方差分析 )…     

Development of an immunochromatographic assay for the rapid detection of bromoxynil in water

A rapid immunochromatographic one-step strip test was developed to specifically determine bromoxynil in surface and drinking water by competitive inhibition with the nano colloidal gold-conjugated monoclonal antibody (mAb). Bromoxynil standard samples of 0.01–10 mg L⁻¹ in water were tested by this method and the visual limit was 0.06 mg L⁻¹. The assay only required 5 min and one-step by dispensing a drop of sample solution onto a strip. Parallel analysis of water samples with bromoxynil showed comparable results from one-step strip test and ELISA. Therefore, the one-step strip test is very useful as a screening method for qualitative detection of bromoxynil in water.     

Comparing differential tolerance of native and non-indigenous marine species to metal pollution using novel assay techniques

Recent research suggests anthropogenic disturbance may disproportionately advantage non-indigenous species (NIS), aiding their establishment within impacted environments. This study used novel laboratory- and field-based toxicity testing to determine whether non-indigenous and native bryozoans (common within marine epibenthic communities worldwide) displayed differential tolerance to the common marine pollutant copper (Cu). In laboratory assays on adult colonies, NIS showed remarkable tolerance to Cu, with strong post-exposure recovery and growth. In contrast, native species displayed negative growth and reduced feeding efficiency across most exposure levels. Field transplant experiments supported laboratory findings, with NIS growing faster under Cu conditions. In field-based larval assays, NIS showed strong recruitment and growth in the presence of Cu relative to the native species. We suggest that strong selective pressures exerted by the toxic antifouling paints used on transport vectors (vessels), combined with metal contamination in estuarine environments, may result in metal tolerant NIS advantaged by anthropogenically modified selection regimes.     

微波消解逐时络合光度法快速测定聚合铝溶液中的总铝含量

聚合铝中的3种铝形态及其总铝含量的传统测定方法分别为Al-Ferron逐时络合光度法和EDTA络合滴定法．这2种方法各自存在着许多不足之处．将微波消解技术应用于逐时络合比色法以便快速测定聚合铝溶液中的总铝含量．研究表明：（1）Ferron混合显色液配制好后存放与否对测定结果的影响很大,用微波辐射处理Ferron混合显色液30s后即可使用,克服了传统光度法中必须存放5d的缺点;（2）微波消解极大地缩短了聚合铝与Ferron之间的解聚／络合反应时间,聚合铝显色溶液微波消解2min即可方便地定量测定铝盐水处理剂中的总铝含量,克服了传统络合滴定法的不足和烦琐．同时用改进的Al-Ferron逐时络合比色法验证了这种总铝测定方法的可行性．