末端限制性酶切片段长度多态性分析（terminal restriction hagment length polymorphism，T-RFLP）是近年来发展起来的、不依赖于培养的微生物群落分析方法之一。由于其在微生物群落结构分析方面的特点，包括分辨率高、易于实现自动化及互联网海量数据共享等优势，自1997年最先被报道以来得到了广泛的应用，成为环境微生物群落分析的最强有力的工具之一。本文详细介绍了T-RFLP技术的原理，并从环境样品群落DNA的提取、引物设计和PCR扩增、限制性酶切、电泳分离检测和T-RFLP图谱解析等5个方面讨论了用该技术解析环境微生物群落的方法和技巧，简述了近8a来国外T_RFLP技术在群落分析中的研究进展。类似于其他的分子微生物生态学技术，T-RFLP也有自身的缺陷，因此重点分析了该技术的局限性及相应的解决办法。

A moderately halophilic bacterium, designated strain SL014B-85T, was isolated from a crude-oil-contaminated saline soil from Shengli oilfield, Shandong Province, China. Cells were Gram-negative, aerobic, short rods with lateral flagella. Growth occurred at NaCl concentrations of 0-15% (optimum 5-15%), at 10-42℃ (optimum 30℃) and at pH 8.0-9.0 (optimum pH 8.5). The only respiratory quinone was Q9, and the main cellular fatty acids were C18: 1v7c, C16: 0 and C19: 0 cyclo v8c. The G+C content of the DNA was 66.6 mol%. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain SL014B-85T belonged to the genus Halomonas in the Gammaproteobacteria, with highest sequence similarity of 98.1 and 97.8% to Halomonas alimentaria DSM 15356T and Halomonas ventosae DSM 15911T, respectively. DNA-DNA relatedness values were below 40% with members of closely related Halomonas species. Results of phenotypic, biochemical and phylogenetic analyses revealed that strain SL014B-85T could be classified as representing a novel species of the genus Halomonas, for which the name Halomonas shengliensis sp. nov. is proposed. The type strain is SL014B-85T (=CGMCC 1.6444T=LMG 23897T).

采用油井采出液培养基和加入无机盐成分的改良油井采出液培养基，对大庆油田萨北过渡带油井采出液中的细菌进行分离培养及初步鉴定，比较了两种情况下培养出的具有硝酸盐和/或亚硝酸盐还原，以及/或反硝化能力菌群结构的差异。利用采出液培养基培养出一组新的微生物菌株，并且分离的硝酸盐和/或亚硝酸盐还原菌，以及/或反硝化细菌（Nitrate/nitrite reducing bacteria, denitrifying bacteria, NRDB）比例明显高于无机盐- 采出液培养基；但培养基中无机盐成分的添加，提高了可培养NRDB的群落生物多样性。仅仅向油井采出液中直接投加硝酸盐作为电子受体，对其中硝酸盐还原、亚硝酸盐还原和反硝化微生物（NRDB）的激活作用以及产抑制硫化物产生的能力有限，而同时加入分离自采出液的NRDB和硝酸盐则对硫酸盐还原菌（SRB）的生长和产硫化物活性都产生了明显的抑制。但是NRDB与硝酸盐同时投加对不同SRB的抑制效果并不相同，导致了SRB群落结构的变化。

受粪便污染的水体可能含有严重威胁人类和动物健康的致病微生物，由于病原微生物在水环境中往往数量较少、检测比较困难，通常采用检测与病原微生物具有密切关系的指示微生物来指示和估计病原污染。指示微生物包括总大肠菌群及其替代微生物，已被广泛应用于水体污染程度检测等方面。总结了各种指示微生物的特性、应用范围以及检测方法等方面的最新研究进展。初步介绍了指示微生物和水环境之间的关系，着重介绍了判断病原微生物污染来源的指示微生物及其最新检测方法。

Two novel strains, SL014B61AT and SL014B11A, were isolated from an oil-polluted saline soil from Gudao in the coastal Shengli Oilfield, eastern China. Cells of strains SL014B61AT and SL014B11A were motile, Gram-negative and rod-shaped. Growth occurred at NaCl concentrations of between 0 and 15% and at temperatures of between 10 and 45 6C. Strain SL014B61AT had Q9 as the major respiratory quinone and C16: 0 (21.2%), C18: 1v9c (20.3%), C16: 1v7c (7.3%) and C16: 1v9c (6.4%) as predominant fatty acids. The G+C content of the DNA was 57.9 mol%. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain SL014B61AT belonged to the genus Marinobacter in the class Gammaproteobacteria. Strain SL014B61AT showed the highest 16S rRNA gene sequence similarity with Marinobacter bryozoorum (97.9%) and showed 97.8% sequence similarity to Marinobacter lipolyticus. DNA–DNA relatedness to the reference strains Marinobacter bryozoorum and Marinobacter lipolyticus was 35.5% and 33.8%, respectively. On the basis of these data, it is proposed that strains SL014B61AT and SL014B11A represent a novel species, Marinobacter gudaonensis sp. nov. The type strain is strain SL014B61AT (=DSM 18066T=LMG 23509T=CGMCC 1.6294T).