基于T-RFLP和因子分析的香蒲根际细菌群落研究

以北京市永定河王平湿地为研究对象,应用末端限制性片段长度多态性(T-RFLP)对王平湿地再生水补水口及其远离补水口的香蒲根际细菌和氨氧化细菌(AOB)的群落结构及多样性变化进行分析,在此基础上结合MiCA3比对、典范对应分析(CCA)和多元统计中因子分析(FA),解析湿地不同空间细菌群落功能特性的变化.结果显示再生水补水口细菌群落各多样性指数均明显低于远离补水口细菌群落,而氨氧化细菌群落则呈现相反趋势.基于传统的T-RFLP片段MiCA3比对、CCA和FA相结合的分析表明:对细菌群落,占再生水补水口细菌群落总丰度55.6%的菌群与150bp代表的鞘氨醇单胞菌(Sphingomonas sp.)具相似净化功能,并与总有机碳生物化学循环过程具密切关系,其次占再生水补水口上游细菌群落总丰度75.5%的菌群与81bp为代表的假单胞菌属(Pseudomonas sp.)和Geitlerinema sp.具相似净化功能,并与氮的生物化学循环过程具密切关系.占再生水补水口下游细菌群落总丰度68.7%的菌群与138bp代表的小单孢菌属(Micromonospora sp.)具相似净化功能,同时与重金属Cu、V、和Ti生物化学循环具密切关系.对氨氧化细菌,266bp受铵态氮影响较大,占再生水补水口氨氧化细菌群落总丰度65.5%的菌群与266bp代表的Nitrosomonas sp.具相似的生境特征,适合在高氨环境中生长;与58bp所代表的Nitrosospira sp.具相似功能特性菌群丰度在远离补水口样点中较前者呈现显著增加趋势,表征该类菌群适合在相对低氨环境中生长.     

污泥负荷对生物处理系统耐药细菌的影响研究

为了研究污水生物处理工艺中抗药性细菌生长和分布特性及污泥负荷的影响,构建了不同处理负荷的活性污泥工艺,并以磺胺嘧啶抗性异养菌为例,阐述了污泥负荷对活性污泥系统中典型抗药细菌的生长及排放特性的影响.结果表明,污泥负荷增大有利于磺胺嘧啶抗性异养菌的生长繁殖,负荷提高后净比生长速率和细菌产量分别由0.32d-1和2.3×106CFU/d提高至0.33d-1和3.1×106CFU/d,活性污泥、出水和剩余污泥中抗药菌的浓度也均显著提高(P < 0.05),但对抗药细菌的相对丰度无显著改变.低污泥负荷下[0.24kg COD/(kg MLSS?d)]抗药细菌主要通过剩余污泥形式排放,排放量比(泥中排放量/水中排放量)为28.4;负荷提高至0.4kg COD/(kg MLSS?d)后,出水抗药细菌排放量显著提高,排放量比为1.1.处理相同水量,高污泥负荷下排放的抗药细菌总量明显降低,提高污泥负荷有利于活性污泥系统抗药性风险的控制.     

聚乙烯醇凝胶包埋固定化细菌联合植物的除氮研究

为了解聚乙烯醇(PVA)凝胶包埋固定化菌在富营养化水体生态修复中的效果,利用PVA凝胶包埋固定硝化和反硝化细菌并联合水芹(Oenanthe clecumbens)和蕹菜(Ipomoea aquatica)2种植物进行21 d的除氮试验。结果表明,加菌试验组TN、NH4+-N、NO3--N、NO2--N及TP去除率显著高于无菌试验组(P0.05);各试验组N的去除效率与添加PVA凝胶包埋固定硝化和反硝化细菌呈显著正相关(P0.05)。添加PVA凝胶固定硝化和反硝化细菌的蕹菜和水芹试验组植物的平均生长速率(RGR)分别为38.5和19.6 mg·g-1·d-1,而单纯蕹菜和水芹试验组RGR分别为29.9和9.5 mg·g-1·d-1。     

光合细菌对2,4,6-三氯苯酚的降解特性研究

研究了混合光合细菌PSB-DR在不同光照、接种量和pH值下对2,4,6-三氯苯酚(2,4,6-TCP)的生物降解特性,确定了PSB-DR生物降解2,4,6-TCP的优化控制条件.结果表明,光照培养下,接种量30%,初始pH值7.0时2,4,6-TCP降解效率最高.在此条件下,50mg/L的2,4,6-TCP经5d后降解率达到82.3%.培养基中醋酸钠的加入对2,4,6-TCP降解有明显的抑制作用.PSB-DR静息细胞对2,4,6-TCP的降解符合高浓度底物抑制的酶促反应类型,其降解动力学参数rmax=1.746h-1,Km=38.333mg/L,Ki=260.87mg/L.     

Application of rotating cylinder method for ecotoxicological evaluation of antifouling paints

The environmental impact of two biocide-free antifouling paints, fluoropolymer and silicone types, painted on a test cylinder was assessed using a battery of ecotoxicity test and chemical analyses for organic micro-pollutants such as perfluoroalkyl substances (PFAS). A biocide paint containing zinc pyrithione (ZnPT₂) and cuprous oxide was assessed as a positive control. A standardized laboratory rotating-cylinder method using each test cylinder with artificial seawater was performed for 45 days. After 1?h rotation, the leaked seawater was subjected for bioassay and chemical analyses twice weekly. The seawater extracts from the biocide paint showed adverse effects on bacteria, algae, and crustaceans, but those from the biocide-free paints did not. The leakage seawater from biocide-free paints, after 7-day continuous rotation, contained the same concentration levels of PFAS as blank seawater used to conduct the tests. Thus, no significant toxicities of the biocide-free paints were found under the conditions of this study. Simultaneous analysis of ZnPT₂ and copper pyrithione (CuPT₂) was developed using an HPLC with a polymeric resin column, showing that ZnPT₂ was converted to CuPT₂ by trans-chelation in the leakage seawater from the positive control paint. The experimental results using a laboratory rotating-cylinder method demonstrated that biocide-free paints did not markedly affect three species tested and no PFAS was detected. In contrast, biocide paint was significantly toxic to test species and toxicity of the extract on bacteria was partly responsible for CuPT₂ produced in leakage seawater. Thus, a laboratory rotating-cylinder method may be applied for ecotoxicological assessment of antifouling paints.     

Genetic toxicology of metal compounds: An examination of appropriate cellular models †

Bacterial systems have not had success predicting metal carcinogenicity. Hypotheses explaining this failure are examined. Using a broad genetic endpoint, λ prophage induction, under sub‐toxic growing conditions, genotoxicity is seen for compounds of chromium, manganese, lead, molybdenum and tungsten. Copper, manganese, arsenic and molybdenum compounds enhanced UV mutagenesis in E. coli WP2.

The toxicity of metal compounds to cultured mammalian cells correlates well with rat oral LD₅₀ values. Whereas insolubility can present problems in bacterial studies, concentrations of metal compounds toxic to mammalian cells can be determined even in the presence of precipitate, and sometimes [Pb(NO₃)₂, BaCl₂ and BeCl₂] occurs only in its presence. PbS and MnS, which are insoluble, are much more toxic than the more soluble compounds Pb(NO₃)₂ and MnCl₂. These results demonstrate the importance of cellular phagocytosis of insoluble metal compounds as a factor in studying the toxicity and genotoxicity of metal compounds.     

Large plasmids governing multiple resistances to heavy metals: A genetic approach∗

Gram negative bacteria classified as Alcaligenes eutrophus and carrying large resistance plasmids (generally two) were found in various industrial sites highly contaminated by heavy metals (Zn⁺⁺, Cu⁺⁺, Co⁺⁺,...). These strains were detected by DNA hybridization with a probe made with a 9kb fragment (ccz⁺ fragment) encoding for resistances to Cd⁺⁺, Co⁺⁺ and Zn⁺⁺, and cloned from plasmid pMOL30. This plasmid was isolated from the representative strain A. eutrophus CH34 which harbours the plasmids pMOL30 (240 kb) and pMOL28 (165 kb). Phenotypes related to pMOL28 and pMOL30 include the tolerance to Cd⁺⁺, Co⁺⁺, Cr0₄ ⁼, Cu⁺⁺, Hg⁺⁺, Ni⁺⁺, Pb⁺⁺ and Zn⁺⁺. The described genetic properties of these plasmids refer to some cloned or mapped functions and to some plasmid rearrangements. Plasmid pMOL85 (250 kb) which is related to pMOL30 was also described. Its host (A. eutrophus DS185) was isolated from a zinc desert. pMOL85 can efficiently self transfer in plasmidfree derivatives.     

多基质时厌氧氨氧化菌、异养反硝化污泥活性及抑制特征

以厌氧氨氧化污泥(AAOB)和各种有机物混合,AAOB分别与异养甲醇反硝化菌和城市污水处理厂活性污泥相混合为研究对象,考察了多种基质(醇类、糖类)时自养、异养反硝化菌群的活性及其相应的抑制特征.结果表明,醇类有机物对AAOB的活性有明显抑制作用,甲醇的抑制性最强,当甲醇为5.48mmol/L时,AAOB活性了损失2/3.而乙酸钠对AAOB有一定的促进作用,另外,葡萄糖、乳糖和蔗糖等糖类有机物对AAOB的影响较小.此外,试验发现经驯化的甲醇反硝化菌能利用各种醇类完成异养短程反硝化.对于活性污泥,利用乙酸钠实现短程反硝化的能力优于甲醇、乙醇、葡萄糖和乳糖.混合试验中,正丙醇导致AAOB和甲醇反硝化菌混合菌群中AAOB活性下降,并且在与甲醇反硝化菌的竞争中处于劣势.乙酸钠对于AAOB和活性污泥混合菌群中AAOB的影响较小.     

湖泊沉积物中氨氧化微生物的amoA基因数量

采用定量PCR方法测定了4个湖泊沉积物中氨氧化微生物的amoA基因数量,并分析了其与环境因子之间的关系. 结果表明:小南湖AOA(氨氧化古菌)和AOB(氨氧化细菌)的amoA基因数量最多,分别达2.1×104和2.8×103copies/g(以干质量计,下同);梁子湖仅检测到了AOA amoA基因的存在,平均值为4.9×103copies/g. 东湖和汤逊湖的AOA amoA基因数量比较接近,约为3.0×103copies/g,然而AOB的amoA基因数量在这2个湖泊中仅分别为37和86copies/g;在这些采样点中,AOA的amoA基因数量是AOB的3~278倍. 统计分析发现,随着湖泊营养水平和间隙水中ρ(NH₄+)的上升,AOA和AOB的amoA基因数量均呈增加趋势,但ρ(NH₄+)增加对AOB的促进作用要大于AOA,导致AOA和AOB的amoA基因数量比值与间隙水中ρ(NH₄+)呈显著负相关. pH上升对2类氨氧化微生物的抑制作用则与ρ(NH₄+)增加对它们的促进作用相反. 沉积物中amoA基因数量与间隙水中ρ(NO₂-)无显著相关性,但与ρ(NO₃-)呈显著正相关. 由于ρ(NH₄+)与ρ(DO)之间呈显著负相关,因此认为ρ(DO)与氨氧化微生物amoA基因数量之间的显著负相关可能更多的是对ρ(NH₄+)与氨氧化微生物amoA基因数量之间紧密关系的一种间接反应.      

Characterisation and identification of carbofuran-utilising bacteria isolated from agricultural soil

A total of 96 bacterial cultures were isolated from soil. Seventeen bacterial isolates were selected following their cultivation on solid media containing 100 mg · L^?1 carbofuran as the sole source of carbon and nitrogen. Of the 17 isolates, 10F, 11M, 17N, 23B and 26M were specifically chosen because of their relatively higher growth efficiency and genetic diversity based on Box-polymerase chain reaction analysis. These bacterial cultures had 16S rRNA gene sequences that were most similar to Acinetobacter baumannii (10F), Agrobacterium tumefaciens (11M), Ochrobactrum anthropi (17N), Escherichia coli (23B) and Agrobacterium tumefaciens (26M) with 97, 95, 93, 95 and 94% similarity in their 16S rDNA gene sequence, respectively. Degradation rates of carbofuran in soil inoculated with these isolates were 1.9, 1.5, 1.6, 1.7 and 1.6 times, respectively, faster in comparison with uninoculated soil after 10 days of incubation. The maximum degradation rates of carbofuran (45 and 91%) were detected in soil inoculated with A. baumannii (10F) after 10 and 20 days’ incubation, respectively. These data indicate that these isolates may have the potential for use in bioremediation of pesticide contaminated soil.