施用城市污泥堆肥对土壤微生物群落结构变化的影响简

在温室条件下进行了15周的盆栽实验,考察了施用城市污泥堆肥后,土壤中养分含量的变化规律,重点研究了施用城市污泥堆肥对土壤微生物群落结构变化的影响。实验发现,污泥堆肥能改善土壤养分,有机质和氮、磷含量得到显著提高。经PCR-DGGE分析,施肥1周后土壤中细菌和真菌的群落结构均发生了较大的变化。随着施肥时间的延长,细菌在富含有机质及氮、磷等养分的土壤环境下大量生长,多样性提高,其优势菌群属于γ变形菌、α变形菌和芽单胞菌;随着有机质的不断消耗,细菌的生长活性受到抑制,最终由于养分的缺乏,细菌种群多样性呈现小幅度的降低,优势菌群变为绿弯菌门、γ变形菌亚纲和厚壁菌门。对于真菌,其多样性指数在堆肥前3周逐渐提升,在第3~12周的监测中呈现相对稳定的变化趋势,优势菌群主要为座囊菌纲和散囊菌纲。     

超声波-缺氧／好氧污泥消化反应器中氨氧化细菌群落结构的演变

为了揭示超声波-缺氧／好氧污泥消化反应器中氨氧化细菌群落结构多样性的演变过程,采用变性梯度凝胶技术（PCR—DGGE）研究不同运行时期氨氧化细菌群落结构的变化。DGGE分析表明,反应器中氨氧化细菌群落比较丰富。在反应器运行的不同时期,氨氧化细菌的群落结构发生了一定的变化,反映了种群的动态演变。UPGMA聚类分析将DGGE图谱分为3大类群并对应于各自的运行时期。测序结果表明,反应器中的优势种群属于变形菌门（Proteobacteria）,包括α-proteobacteria,β-proteobacteria,γ-proteobacteria和ε-proteobacteria4个纲,其中β-proteobacteria占45％,γ-proteobacteria占40％。在始终保持明显优势地位的种属中,5株为反硝化细菌,它们对提高反应器脱氮效率具有重要作用。     

城市污泥堆肥产品施用对沙荒地土壤理化性质及高羊茅生长的影响

通过盆栽实验研究了城市污泥堆肥产品4种施用比例(0、10%、25%和50%)对沙荒地土壤理化性质及高羊茅生长的影响,结果表明:污泥堆肥产品的施加使混配土壤p H值向更利于高羊茅生长的p H范围(6.0~7.0)变化,土壤电导率随着施肥比例的增大有显著的增加,当施肥比例为10%、25%、50%时,土壤电导率分别为不施肥沙土的11.93、12.66和19.81倍;城市污泥堆肥产品施用后可显著提高沙荒地土壤中全氮、全磷、全钾及有机质含量,施肥比例为10%、25%和50%的沙土中有机质含量分别是不施肥沙土的3.69、4.22和5.65倍。污泥堆肥产品的施入有利于高羊茅对营养元素氮、磷和钾的吸收,进而促进了高羊茅的生长,但是这种促进作用在实验初期较为缓慢,直到实验后期方才显现出来。当施肥比例为25%时,高羊茅对营养物质的利用率达到最高,确定其为沙荒地土壤改良较为适宜的施肥比例。     

污泥施用对根际和非根际石灰性土壤中细菌多样性的影响

为研究污泥施用对石灰性土壤细菌多样性的影响,在连续2年种植夏玉米大田系统中,施用不同量的腐熟污泥,采集根际和非根际土壤进行16S rDNA-V3-V4区高通量测序,分析污泥施用对石灰性土壤细菌丰富度、多样性指数、群落结构、功能基因在代谢途径的影响,同时对细菌群落和环境因素之间的相关性进行分析。结果表明:污泥用量分别在3.5～37.5 t·hm~(-2)和3.5～7.5 t·hm~(-2)有利于非根际和根际土壤OTU数量增加;但污泥在75 t·hm~(-2)过量施用时,根际和非根际土壤细菌的Shannon多样性指数显著降低,Simpson指数显著增加,Ace,Chao丰富度指数显著降低。主成分和门水平物种丰度分析表明:污泥施用量不同会造成根际和非根际细菌群落结构差异,并且当污泥施用量为75 t·hm~(-2)时,明显降低了非根际土壤放线菌门的丰度;污泥施用量在3.75～37.5 t·hm~(-2)时,非根际土壤中拟杆菌门丰度会明显增加。与对照相比,当污泥用量为75 t·hm~(-2)时,根际土壤拟杆菌门有明显的增加,增加丰度达1.45倍,但酸杆菌门和放线菌门丰度降低了49.74%和80.57%。冗余分析(RDA)表明,土壤TN、Cd、Cu、TP是影响土壤细菌群落最主要的因素,其中TN、Cd、Cu和细菌Shannon、Simpson多样性指数呈现显著相关性(P0.05)。由此可见,连续过量施入污泥会对根际和非根际石灰性土壤中细菌多样性造成不利影响,而且上述微生物多样性变化可作为污泥合理施用的判断依据。     

ABR-MBR工艺反硝化除磷微生物群落特征分析

为了揭示ABR-MBR组合工艺中反硝化除磷微生物种群演替规律,采用Miseq高通量测序技术考察了该工艺在不同运行阶段除磷功能区的微生物群落结构。结果表明,硝化液回流比逐步从150%提升至300%可促进反硝化除磷菌大量富集,促进系统的启动和稳定运行;系统在运行过程中始终保持较高的微生物多样性;优势微生物种群均以变形菌门(Proteobacteria)和拟杆菌门(Bacteroidetes)为主,最大丰度分别为55.13%和7.76%,且变形菌门功能性微生物主要集中在γ-变形菌纲(Gamaproteobacteria);功能性除磷菌属主要为气单胞菌属(Aeromonas),假单胞菌属(Pseudomonas)和黄杆菌属(Flavobacterium);其中在逐步提升硝化液回流比过程中气单胞菌属(Aeromonas)被大量富集,其在γ-变形菌纲(Gamaproteobacteria)的相对丰度由5.30%上升至41.49%并在系统后续运行中维持主导地位。系统除磷效果与功能性除磷微生物相对丰度的变化密切相关。系统中微生物种群的多样性和功能微生物的结构稳定性为ABR-MBR工艺的稳定运行和高效处理提供了保证。     

饮用水管网生物膜细菌群落特征及其对腐蚀的影响

通过454高通量测序对北方A和B两实际饮用水管网系统中生物膜细菌群落特征进行表征,并研究了其对管网腐蚀产物组成的影响。结果表明,A管网生物膜细菌群落丰度和多样性高于B管网生物膜。在门水平上,两管网生物膜细菌群落主要为变形菌门Proteobacteria,但B管网生物膜中的相对丰度(67.10%)高于A(46.50%)。在纲水平上,A管网生物膜主要为β-变形菌纲Betaproteobacteria,B管网生物膜中主要为β-变形菌纲Betaproteobacteria和δ-变形菌纲Deltaproteobacteria。在属水平上,A管网生物膜中腐蚀相关菌群主要为硝酸盐还原菌和铁还原菌,其腐蚀产物主要为α-Fe OOH和Fe_3O_4,而B管网生物膜中硫酸盐还原菌特别是脱硫弧菌属Desulfovibrio含量较高,其可能与管垢中绿锈含量高有关。     

外源菌剂对餐厨垃圾和污泥联合堆肥酶活性及微生物的影响

以餐厨垃圾和污泥(质量比1∶1)混合物为底物,以不接种微生物为对照,以添加外源菌剂和腐熟堆肥为两个处理,研究外源菌剂对联合堆肥过程中酶活性和腐熟期微生物的影响。结果表明,外源菌剂可促进堆体升温并延长高温期,可显著提高纤维素酶、过氧化氢酶、脲酶、蛋白酶的活性,4种酶的峰值分别为377.294 U/g、83.107 mL/(g·h)、0.763 mg/(g·d)和147.411 U/g。采用高通量测序技术分析发现,添加外源菌剂降低了腐熟期细菌、真菌的群落多样性,但提高了真菌群落丰富度。门水平上,各堆体细菌和真菌的优势群落种类差异较小,仅相对丰度变化较大;属水平上,添加外源菌剂使堆体中优势细菌占比更均匀,优势真菌主要为土壤伊萨酵母菌(Issatchenkia)、念珠菌(Candida)、未分类真菌(Unclassified Fungi)等。采用FAPROTAX工具对细菌功能分组,各堆体相对丰度较高的功能菌以化能异养、有氧化能异养、硝酸盐还原等为主,然而添加外源菌剂和腐熟堆肥使芳香族化合物的降解功能菌相对丰度分别提高了8.0倍、7.4倍。     

陕北石油污染区土壤细菌群落结构解析

为了解陕北地区石油污染对土壤微生物群落结构的影响,采用高通量测序对石油污染土壤中细菌群落结构进行研究,考察了石油污染土样的理化性质、可培养微生物数量及细菌群落结构,探讨石油污染程度与细菌群落结构间的关系。结果表明,石油污染土样中残油量为28.7~870.0mg/kg,相对于未污染土样,石油污染土样的总碳随残油量的增加呈增加趋势。石油污染土样中可培养细菌数量大于未污染土样,而可培养放线菌数量则相反。细菌群落结构分析表明,石油污染土样中细菌群落数量显著低于未污染土样,操作分类单元(OTU)数从未污染土样的7 707个最低下降到2 034个,且物种丰富度显著降低,其中厚壁菌门、放线菌门及变形菌门为优势菌群,随土壤残油量增加,厚壁菌门、变形菌门及拟杆菌门比例显著上升,而酸杆菌门、芽单胞菌门等比例则显著降低。     

规模化奶牛养殖粪水还田对土壤理化性质及微生物群落特征的影响

规模化奶牛养殖粪水还田可实现资源化利用,但其对土壤环境的影响尚待明晰。通过完全随机区组设计试验研究了粪水还田量分别为0、0.32、0.64、0.96 t/m²时对土壤理化性质及微生物群落特征的影响。结果表明：养殖粪水还田可以提高土壤养分和作物生物量,最合适的粪水还田量应为0.64 t/m²,此时对土壤微生物的丰富度和多样性影响也最小。土壤细菌中的优势细菌属斯克尔曼氏菌属(Skermanella)可生物固氮,减少土壤氮素流失;芽孢杆菌属(Bacillus)能分泌纤维素酶,有利于土壤中纤维素类的降解。子囊菌门(Ascomycota)为最主要的优势真菌门,它们有助于植物纤维素和木质素的降解。     

抗生素菌渣堆肥进程中微生物群落的变化

将青霉素菌渣、林可霉素菌渣与牛粪等原料分别进行好氧堆肥实验,以考察堆肥过程中不同菌渣对微生物群落的影响。在堆制的41d里,根据温度变化分阶段采集堆肥样品,采用稀释倒平板法测定细菌、放线菌和真菌的数量。结果表明,菌渣不同,其堆肥中的微生物群落变化趋势不同。青霉素菌渣堆肥中细菌数量变化趋势为高一低,真菌数量变化趋势为高一低．高,放线菌数量为逐渐增加;林可霉素菌渣堆肥过程中细菌数量变化趋势为低一高一低,放线菌和真菌数量变化趋势为高．低．高。依据真菌菌落形态观察,菌渣堆肥中的真菌种类比对照牛粪堆肥单一,表明两种菌渣对堆肥中的微生物多样性均产生了不利影响。林可霉素菌渣堆肥初始时的细菌数量比对照低1个数量级,放线菌数量在整个堆肥进程中都明显低于对照,堆肥结束时,随着菌渣含量的增加,放线菌数量逐渐下降,高温期真菌数量下降幅度随着菌渣含量增加而加大,表明林可霉素菌渣对细菌、放线菌和真菌均有不同程度的抑制。堆肥化后菌渣中林可霉素残留量的减少表明,在一定条件下堆肥处理可以将抗生素菌渣无害化和资源化。     

Revegetation of lagoon ash using the legume species Acacia auriculiformis and Leucaena leucocephala

A greenhouse study was conducted to evaluate the potential use of two legume species, Acacia auriculiformis and Leucaena leucocephala for growth on ameliorated lagoon ash with or without nitrogen (N(2))-fixing bacteria inoculation. Even though amendments of 30% (w/w) vermiculite or with sewage sludge compost were added to improve the chemical and physical limitations of lagoon ash, significant suppressions in biomass and plant nutrient content were found with ameliorated lagoon ash in comparison to an agricultural soil. The high proportion of clay-sized (<53 microm) ash particles limited root growth. In addition, heavy metal toxicity was a possible factor contributing to poor seedling growth. Higher plant productivity resulted from the sewage sludge compost-amended lagoon ash than with vermiculite due to a greater contribution of plant nutrients in the compost. Nodulation was inhibited in ameliorated lagoon ash but not in agricultural soil. High pH and electrical conductivity and elevated toxic metals may be important parameters that limit bacterial activity. Both species showed potential to establish on amended lagoon ash, with Acacia auriculiformis being the best adapted.     

Carbon/nitrogen ratio as a major factor for predicting the effects of organic wastes on soil bacterial communities assessed by DNA-based molecular techniques

Background, aim, and scope  

Current Australian legislation permits the beneficial application of grease trap waste (GTW) to agricultural soil, viewing it as a beneficial source of organic matter and soil conditioner containing no/low amounts of metals or pathogenic organisms. However, little is known about the influence of GTW on soil bacterial community. A field experiment was established at Menangle in south western Sydney in Australia to quantitatively assess the impacts of different types (GTW CO and GTW CL) and amounts of GTW application on the soil bacterial community and diversity. Furthermore, a municipal solid waste (MSW) compost was simultaneously examined to compare against the other organic wastes. Knowledge about the shifts in microbial community structure and diversity following the applications of organic wastes could help to evaluate the ecological consequences on the soil and thus to develop sound regulatory guidelines for the beneficial reuse of organic wastes in agricultural lands.     

有机碳源条件下厌氧氨氧化SBBR中的微生物多样性

研究有机碳源对SBBR厌氧氨氧化菌群等微生物的影响。采用16S rDNA序列与PCR-DGGE分析技术相结合的方法,对稳定运行的反应器内的活性污泥和生物膜样品,进行细菌多样性图谱分析,同时采用巢式PCR-DGGE技术对浮霉状菌属(Planctomycetes)细菌进行分析。结果表明,在有机碳源反应系统细菌条带数和多样性指数均高于无机系统,与活性污泥相比,生物膜表尤为明显。当进水不含有机碳源时,氨氧化细菌(ammonia oxidizing bacteria,AOB),厌氧氨氧化菌(anaerobic ammonia oxidizing bacteria,ANAMMOX)为优势功能菌;当进水含有机碳源时,系统中存在的AOB以亚硝化单胞菌(Nitrosomonas sp.)为优势菌群,同时存在反硝化菌,如索氏菌(Thauera sp.)以及厌氧氨氧化菌,它们共同作用完成N的去除。此外,与无机碳源系统相比,有机碳源的存在,有利于浮霉状菌的积累,但压缩了ANAMMOX的生存空间。本研究可为厌氧氨氧化工艺处理低C/N比有机废水提供了理论依据。     

The impact of sewage sludge compost on tree peony growth and soil microbiological,and biochemical properties

In order to assess the suitability of sludge compost application for tree peony (Paeonia suffruticosa)–soil ecosystems, we determined soil microbial biomass C (C_mic), basal respiration (R_mic), enzyme activities, and tree peony growth parameters at 0–75% sludge compost amendment dosage. Soil C_mic, R_mic, C_mic as a percent of soil organic C, enzyme (invertase, urease, proteinase, phosphatase, polyphenoloxidase) activities, and plant height, flower diameter, and flower numbers per plant of tree peony significantly increased after sludge compost amendment; however, with the increasing sludge compost amendment dosage, a decreasing trend above 45% sludge compost amendment became apparent although soil organic C, total Kjeldahl N, and total P always increased with the sludge compost amendment. Soil metabolic quotient first showed a decreasing trend with the increasing sludge compost application in the range of 15–45%, and then an increasing trend from compost application of 45–75%, with the minimum found at compost application of 45%. As for the diseased plants, 50% of tree peony under the treatment without sludge compost amendment suffered from yellow leaf disease of tree peony, while no any disease was observed under the treatments with sludge compost application of 30–75%, which showed sludge compost application had significant suppressive effect on the yellow leaf disease of tree peony. This result convincingly demonstrated that ?45% sludge compost application dosage can take advantage of beneficial effect on tree peony growth and tree peony–soil ecosystems.     

水解溶菌酶污泥减量过程中的微生物群落变化

利用水解溶菌酶对SBR系统中的剩余污泥进行减量。通过与未加水解溶菌酶的相同系统对比,研究了水解溶菌酶作用下的SBR系统中剩余污泥的减量效果与微生物群落结构的变化。结果表明,在50 d的运行期内,水解溶菌酶作用下的SBR系统中剩余污泥减量总计达到76.3%,同时该系统对COD与TN的平均去除率分别为88.2%与53.8%。通过PCR-DGGE分析可知,随着运行时间的增加两系统微生物群落结构的差异逐步明显,SBR系统中原有的部分优势微生物在水解溶菌酶的作用下逐渐减弱。另外,对微生物群落的部分优势细菌进行克隆测序和系统发育树分析,通过鉴定获得的7条细菌的16S rDNA序列,它们分别与放线菌和杆菌同源性在97%以上。     

Examination of environmental quality of raw and composting de-inking paper sludge

Paper sludges were traditionally landfilled or burned. Over the years, the use of paper sludges on soils has increased, as well as the concerns about their environmental effects. Therefore, the chemical characterization of paper sludges and their young (immature) compost needed to be investigated, and over 150 inorganic and organic chemicals were analyzed in de-inking paper sludge (DPS). In general, nitrogen, phosphorus and potassium contents were low but variable in raw DPS and its young compost. The contents of arsenic, boron, cadmium, cobalt, chromium, manganese, mercury, molybdenum, nickel, lead, selenium, and zinc were also low and showed low variability. However, the copper contents were above the Canadian compost regulation for unrestricted use and required a follow-up. The fatty- and resin acids, and polycyclic aromatic hydrocarbons were the organic chemicals measured at the highest concentrations. For resinic acids, care should be taken to avoid that leachates reach aquatic life. For polycyclic aromatic hydrocarbons, naphthalene should be followed until soil content reaches 0.1 microg g(-1), the maximum allowed for soil use for agricultural purposes according to Canadian Environmental Quality Guidelines. In young compost, the concentration of these chemical families decreased over time and most compounds were below the detection limits after 24 weeks of composting. In raw DPS, among the phenol, halogenated and monoaromatic hydrocarbons, dioxin and furan, and polychlorinated biphenyl families, most compounds were below the detection limits. The raw DPS and its young compost do not represent a major threat for the environment but can require an environmental follow-up.     

Effect of flocculant on microbial populations and function in Chem-Bioflocculation (CBF) treatment process using PCR-DGGE

The group-specific primers were applied to investigate microbial communities in Chem-Bioflocculation (CBF) reactor by PCR-DGGE method. α-proteobacteria, β-proteobacteria, actinomycetes and acidobacterium, were analysed by using diversity index at different dosage of Polyaluminium Ferric Chloride (PAFC) based on DGGE patterns. Clustering analysis showed that community structure of specific bacteria groups changed when flocculant dosage was adjusted, but flocculant dosage had dissimilar effect on the community structure of different bacteria group. α-proteobacteria possibly was the most influential populations relative to biological function in CBF reactor. The biological function should be inhibited when the flocculant dosage exceeded 80 mg/L.     

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.     

Effect of Sphingobium yanoikuyae B1 inoculation on bacterial community dynamics and polycyclic aromatic hydrocarbon degradation in aged and freshly PAH-contaminated soils

Sphingobium yanoikuyae B1 is able to degrade a range of polycyclic aromatic hydrocarbons (PAHs) and as a sphingomonad belongs to one of the dominant genera found in PAH-contaminated soils. We examined the ecological effect that soil inoculation with S. yanoikuyae B1 has on the native bacterial community in three different soils: aged PAH-contaminated soil from an industrial site, compost freshly contaminated with PAHs and un-contaminated compost. Survival of S. yanoikuyae B1 was dependent on the presence of PAHs, and the strain was unable to colonize un-contaminated compost. Inoculation with S. yanoikuyae B1 did not cause extensive changes in the native bacterial community of either soil, as assessed by denaturing gel electrophoresis, but its presence led to an increase in the population level of two other species in the aged contaminated soil community and appeared to have an antagonistic affect on several members of the contaminated compost community, indicating niche competition.