添加猪粪对不同施肥历史土壤细菌群落的影响

为探讨粪肥携带的微生物对土壤微生物的影响，以长期施用粪肥和化肥的土壤为研究对象，耦合传统微生物学培养方法和现代分子微生物学方法，分析了猪粪和灭菌猪粪对2种土壤的细菌数量、群落结构和热代谢活性的影响.结果表明，以9.0t/hm²的用量施入时，猪粪和灭菌猪粪对两种土壤中的可培养细菌数量以及细菌16S-rRNA拷贝数均无明显影响；土壤中的土著微生物对猪粪携带的外源细菌在土壤中定殖具有明显的抑制作用，将2种土壤灭菌后添加猪粪，可培养细菌平均数量较未灭菌土壤分别增加45.96和96.21倍.粪肥输入的化学物质未导致土壤细菌群落结构和相对丰度发生明显变化；而粪肥输入的外源细菌Marinilabiaceae、Porphyromonadaceae、Bacteroidale和Pseudomonadaceae则可以在长期施用化肥的土壤中定殖并对相应的细菌丰度造成影响，但是在长期施用粪肥的土壤中，这些细菌在最开始就被抑制.添加粪肥提高了长期施用化肥土壤的微生物热代谢活性，但对长期施用粪肥的土壤无明显影响.长期施用化肥的土壤受粪源细菌的影响较大，而长期施用粪肥的土壤对粪肥的敏感度降低，对粪肥携带的外源细菌的抑制作用增强.     

粪肥和有机肥施用对稻田土壤微生物群落多样性影响

为探究典型粪肥施用对稻田土壤微生物的影响,开展崇明岛稻田粪肥施用现场实验.采用高通量测序技术分析对照组(CK)和鸡粪(CM)、猪粪(PM)和有机肥(OF)施用稻田土壤微生物群落组成及多样性.结果表明,与CK相比,施用有机肥可提高土壤有机质(SOM),施用鸡粪可显著提高土壤氨氮(NH~+_4-N)和总氮(TN)含量(P0.05).PM组土壤微生物多样性显著高于CK组(P0.05),OF组土壤微生物群落丰富度显著高于CM组(P0.05).pH值、总磷(TP)、总氮和Pb是影响稻田土壤微生物群落组成的重要因素,CM组微生物群落结构与其他3组差异较大.与CK相比,OF组增加了硝化螺菌属(Nitrospira)的相对丰度,CM组显著降低了反硝化细菌Ignavibacteriae的相对丰度(P0.01),达40.56%,但显著增加了硝化细菌陶厄氏菌属(Thauera)的相对丰度(P0.05),达203.00%; PM组显著增加了氨化细菌Armatimonadetes的相对丰度(P0.05),达57.51%,还增加了厌氧绳菌属(Anaerolinea)的相对丰度,达102.00%.施用鸡粪和猪粪分别显著增加致病菌假单胞菌属(Pseudomonas)和黄杆菌属(Flavisolibacter)的相对丰度(P0.05),而有机肥施用则降低了黄杆菌属的相对丰度.粪肥的施用增加了参与稻田土壤氮循环过程细菌的丰度,对调节稻田土壤氮平衡起着正向作用,然而鸡粪和猪粪的直接施用会导致病原菌增多,对稻田土壤健康有一定的胁迫.     

畜禽粪肥还田四环素类抗生素(TCs)在土壤-蔬菜系统的分布特征及风险评估

畜禽粪肥还田引起的土壤抗生素污染对蔬菜安全和土壤生态风险的影响应得到足够的重视.四环素类抗生素(TCs)在畜禽粪肥和蔬菜基地土壤中被普遍检出且含量较高.然而,目前针对土壤TCs污染对蔬菜安全和土壤生态风险的研究主要采用盆栽试验和田间调查的研究方法,较少开展田间试验.结合盆栽试验和田间试验的研究方法,调查畜禽粪肥还田引起的TCs在土壤-蔬菜系统中的分布特征,采用日允许摄入量-危害商值法(ADI-HQ法)和物种敏感性分布-风险商值法(SSD-RQ法)分别对蔬菜可食部分TCs污染的人体健康风险和土壤TCs污染的生态风险进行评估.结果表明,盆栽试验和田间试验菜心地上部和地下部TCs含量均以OTC为主;盆栽试验和田间试验菜心地上部(以干重计)ω(OTC)分别高达29.25μg·kg^-1和45.03μg·kg^-1,地下部含量(以干重计)ω(OTC)分别高达87.32μg·kg^-1和135.44μg·kg^-1;且田间试验菜心地上部和地下部3种TCs含量均显著高于盆栽试验.土壤TCs的残留以OTC为主,盆栽试验和田...     

哈尔滨市蔬菜基地四环素类抗生素的污染现状

该文选取高寒地区哈尔滨市为研究对象,建立了超声波提取、固相萃取(SPE)、液相色谱-紫外检测(LC-UV)结合质谱检测(MS)法,检测了哈尔滨4个主要蔬菜基地(双城、五常、呼兰、平房)土壤中4种四环素类抗生素(四环素、金霉素、土霉素、强力霉素)的残留。质谱检测法证明4种抗生素在土壤样品中均有检出,其中双城蔬菜基地土壤中残留抗生素种类最多,分别为四环素、金霉素和土霉素。液相色谱定量分析结果显示残留抗生素的浓度范围为15.68～98.64μg/kg,其中检出量最高的是平房蔬菜基地土壤中残留的强力霉素。结果表明,哈尔滨4个蔬菜基地土壤中均存在四环素类抗生素的残留。     

北京蔬菜地土壤中抗生素抗性基因与可移动元件的分布特征

为揭示北京地区蔬菜土壤中抗生素抗性基因与可移动元件的分布特征应用高通量荧光定量PCR方法(HT-qPCR),选取北京3个区5个蔬菜基地进行调查研究.在蔬菜基地土壤中共检测到92～121种抗生素抗性基因,4～6种可移动元件,抗生素抗性基因及可移动元件按区分开.各蔬菜基地中共有且丰度较高的抗生素抗性基因型为:多重耐药类oprD、acrA-04和acrA-05,大环内酯类-林肯酰胺类-链阳性菌素B类抗生素抗性基因(MLSB)酰胺酶类fox5,万古霉素类vanC-03;共有可移动元件为intI1.蔬菜基地土壤中共检测到7种抗生素,含量较高的抗生素种类为恩诺沙星(ENR)、诺氟沙星(NOR)、土霉素(OTC)、磺胺甲噁唑(SMX).顺义区S1与S2蔬菜基地土壤中抗生素的种类与丰度均最高,依次是通州区T蔬菜基地、昌平区C2与C1蔬菜基地.相关性分析表明,蔬菜基地土壤中抗生素抗性基因丰度与抗生素丰度存在显著正相关(P 0.05).研究结果可为后续控制抗生素抗性基因的传播提供基础理论数据.     

北京地区菜田土壤抗生素抗性基因的分布特征

为研究北京地区菜田土壤抗生素抗性基因(antibiotic resistance genes,ARGs)的污染状况和分布特征,采集了11个长期施用粪肥蔬菜基地的温室土壤和大田土壤,进行了抗生素以及ARGs种类和丰度的检测.结果表明,菜田土壤中四环素类抗生素残留量最高,其次为磺胺类抗生素,温室土壤抗生素残留普遍高于大田土壤.温室和大田土壤磺胺类抗性基因sul1、sul2和四环素类抗性基因tet L的检出率均为100%.温室土壤的Ⅰ类整合子(int I1)检出率比大田土壤高1.5倍.定量PCR的检测结果表明,菜田土壤中sul2和tet L的相对丰度介于10-4~10-2之间,温室土壤sul2和tet L的相对丰度普遍高于大田土壤.sul2的相对丰度与磺胺二甲嘧啶和强力霉素的含量显著正相关(P0.05),tet L相对丰度与抗生素含量无明显相关性(P0.05).本研究结果对于掌握北京地区农田土壤ARGs的污染现状,以及从ARGs角度评估农艺措施具有重要的指导意义.     

长期施用粪肥土壤中喹诺酮类抗生素的含量与分布特征

针对珠江三角洲地区长期施用粪肥的“无公害蔬菜”生产基地,利用固相萃取-高效液相色谱-串联质谱(LC-MS/MS)分析方法,探讨了土壤中喹诺酮类(QNs)抗生素4种化合物的含量与分布特征.结果表明,土壤中4种化合物的总含量(ΣQNs)为3.97~32.03μg/kg,平均17.99μg/kg.各化合物的检出率除了洛美沙星(92%)以外均为100%,最高含量为17.21μg/kg,平均含量为0.17~9.31μg/kg,以环丙沙星为主,其次是诺氟沙星和恩诺沙星.化合物的含量与组成特征无论在平面上还是在剖面上均存在明显的空间分布差异,且随着深度增加含量迅速降低,但在80cm深度仍有检出.珠江三角洲地区长期施用粪肥蔬菜基地土壤中喹诺酮类抗生素含量较低,土壤生态毒性风险较小,但各化合物均很高的检出率而导致的协同生态毒性尤其是耐药性问题值得进一步研究.     

长期施用粪肥蔬菜基地蔬菜中典型抗生素的污染特征

利用高效液相色谱-串联质谱(HPLC-MS/MS)技术,分析了珠三角地区某长期施用粪肥蔬菜基地蔬菜中16种典型抗生素的含量,并初步评价其健康风险.结果表明,绝大部分抗生素不同程度被检出,检出率为11%～100%,大多数含量<5μg·kg-1(干重),最高为23.88μg·kg-1,平均为0.91μg·kg-1,以恩诺沙星、环丙沙星、磺胺二甲基嘧啶和磺胺嘧啶为主.各蔬菜样品中至少检出1种抗生素,最多达10种.蔬菜不同器官中喹诺酮类抗生素含量为地下部>地上部.人体通过食用研究区蔬菜摄入抗生素的量低于日允许摄入量(ADI),表明其健康风险较小,但多种抗生素的联合毒性及其耐药性问题不容忽视.     

动物粪便施肥措施促进耐药基因在粪便-土壤-蔬菜之间的散播

耐药基因（antibiotic resistant gene,ARG）在动物粪肥施用的土壤中被越来越多地检出,已经引起了人们对公共安全的担忧,但目前关于动物粪便施肥对蔬菜内生菌中耐药基因的影响尚不清楚.因此,本研究利用高通量定量PCR技术,探讨鸡粪肥施用对土壤、苦菊根和叶片中内生细菌菌群和耐药基因谱的影响.研究结果发现,鸡粪肥的施用不仅增加了土壤和苦菊根内生菌中ARG的数量,而且增加了土壤、苦菊根和叶片中内生菌的ARG丰度.相关性分析显示,土壤菌群和苦菊内生菌中ARG谱与细菌菌群组成密切相关,主要涉及变形杆菌纲、酸杆菌纲、放线菌纲和蓝藻细菌纲等细菌菌群.此外,苦菊内生菌与土壤菌群之间存在共有的ARG,表明土壤-苦菊之间存在耐药基因的内部传播.综上所述,鸡粪肥施用会通过粪便-土壤-植物路径增加蔬菜内生菌中ARG的多样性和丰度.     

施用不同来源粪肥对土壤中抗生素淋溶的影响

含有抗生素的粪肥进入农田土壤中对土壤生态系统和水体环境产生危害,同时也会造成抗生素耐药性的传播,对环境安全和人体健康存在威胁.本研究采用土柱淋溶法,以添加四环素类抗生素（四环素、土霉素和金霉素）的猪粪（PM-T）、牛粪（CM-T）、鸡粪（CHM-T）及空白处理（CK-T）为实验组,未添加抗生素的处理为对照组,研究了模拟淋溶条件下施用不同来源粪肥对土壤理化指标和细菌丰度的影响,阐述不同处理四环素类抗生素的迁移规律.结果表明,相比于CHM-T和CM-T处理组,四环素类抗生素更容易在PM-T处理中累积,残留量为0.90~6.91 mg·kg^-1,明显高于其他处理（0.33~4.42 mg·kg^-1）;相比于表层土壤（0~4 cm）,四环素类抗生素在下层土壤（16~24 cm）的浓度更高.PM-T处理土壤中TN和NH₄⁺-N含量分别增加44 mg·kg^-1和14.11 mg·kg^-1,明显高于其他处理.在抗生素的抑制作用下土壤中细菌丰度减少,PM-T处理土壤中的细菌丰度降低39.66%,高于其他添加抗生素处理组（12.38%~35.26%）.CHM-T处理中抗生素更容易从土壤中淋出,淋出总量为9.91 mg,明显高于其他粪肥处理组（P<0.05）.TN、NH₄⁺-N、四环素、土霉素和金霉素等指标为第一主成分因子,解释量为54.55%,其土壤中的含量与土层深度正相关.总结得出,以猪粪为原料的粪肥处理中四环素类抗生素更容易累积在土壤中,特别是土壤下层,同时更容易造成土壤中微生物环境的改变;而以鸡粪为原料的粪肥施用后抗生素更容易淋出土壤,增加水体污染的风险.     

Effects of two sludge application on fractionation and phytotoxicity of zinc and copper in the soil

The potential harm of heavy metals is a primary concern in application of sludge to the agricultural land. A pot experiment was conducted to evaluate the effect of two sludges on fractionation of Zn and Cu in soil and their phytotoxicity to pakchoi. The loamy soil was mixed with 0%, 20%, 40%, 60% and 80% (by weight) of digested sewage sludge (SS) and composted sludge (SC). The additions of both sludges caused a significant raise in all fractions, resulting in that exchangeable (EXCH) and organic bound (OM) became predominance of Zn and organic bound Cu occupied the largest portion. There was more available amount of Zn and Cu in SS treatments than SC treatments. During the pot experiment, the concentration of Zn in EXCH, carbonate (CAR) and OM and Cu in EXCH and OM fractions decreased in all treatments, so their bioavailability reduced. Germination rate and plant biomass decreased when the addition rate was high and the best yield appeared in 20% mixtures at the harvest of pakchoi. The two sludges increased tissue contents of Zn and Cu especially in the SS treatments. Zn in pakchoi was not only in relationship to ΔEXCH and ΔCAR forms but also in ΔOM forms in the sludge-soil mixtures. Tissue content of Cu in pakchoi grown on SC-soils could not be predicted by ΔEXCH. These correlation rates between Zn and Cu accumulation in pakchoi and variation of different fractions increased with time, which might indicate that sludges represented stronger impacts on the plant in long-term land application.     

Intraspecific differences in effects of co-contamination of cadmium and

A hydroponic experiment was carried out to study intraspecific differences in the effects of different concentrations of cadmium (Cd)(0-10 mg/L) and arsenate (As(V)) (0-8 mg/L) on the growth parameters and accumulation of Cd and As in six wheat varieties Jing-9428, Duokang-1, Jingdong-11, Jing-411, Jingdong-8 and Zhongmai-8. The endpoints of wheat seedlings, including seed germination,biomass, root length and shoot height, decreased with increasing the Cd and As concentrations. Significant differences in seed germination, biomass, root length, shoot height and the accumulation of Cd and As were observed between the treatments and among the varieties (p ＜ 0.05). The lethal dosage 50% were about 20, 80, 60, 60, 80 and 20 mg As/L for Jing-9428, Duokang-1, Jingdong-11,Jing-411, Jingdong-8 and Zhongmai-8, respectively, and the corresponding values for Cd were about 30, 80, 20, 40, 60 and 10 mg Cd/L, respectively. Among the six varieties, Duokang-1 was found to be the most resistant to Cd and As toxicity, and Zhongmai-8 was the most sensitive to Cd and As co-contamination. The resistance of the six varieties was found dependant on the seedling uptake of Cd and As. Duokang-1 was the most suitable for cultivation in Cd and As co-contaminated soils.     

EDTA-enhanced phytoremediation of lead contaminated soil by Bidens maximowicziana

Phytoremediation is a potential cleanup technology for the removal of heavy metals from contaminated soils.Bidens maximowicziana is a new Pb hyperaccumulator,which not only has remarkable tolerance to Pb but also extraordinary accumulation capacity for Pb.The maximum Pb concentration was 1509.3 mg/kg in roots and 2164.7 mg/kg in overground tissues.The Pb distribution order in the B. maximowicziana was:leaf>stem>root.The effect of amendments on phytoremediation was also studied.The mobility of soil Pb and the Pb concentrations in plants were both increased by EDTA application.Compared with CK(control check),EDTA application promoted translocation of Pb to overground parts of the plant.The Pb concentrations in overground parts of plants was increased from 24.23-680.56 mg/kg to 29.07-1905.57 mg/kg.This research demonstrated that B.maximowicziana appeared to be suitable for phytoremediation of Pb contaminated soil,especially,combination with EDTA.     

Distribution and ecological effect of mercury in Laogang landfill, Shanghai, China

Laogang landfill near Shanghai is the largest landfill in China, and receives about 10000 t of daily garbage per day, Samples of topsoil and plants were analyzed to evaluate mercury pollution from the landfill. For topsoil samples, there were significant correlations among total mercury （HgT）, combinative mercury （Hgc） and gaseous mercury （HgG）, and content of total organic carbon （TOC）, but, no significantly relationship was found between Hg content and filling time. Hg content changes in vertical profiles with time showed that the average Hgv of profiles 1992, 1996, and 2000 was similar, but their average HgG was quite different. HgT was significantly correlated with Hgc in profile 1992 and 2000, and Hgv was significantly correlated with Hg6 in profile 1996. HgG/Hgv ratio in profile samples decreased in the order of （HgG,/HgT）1992〉（HgG/HgT）1996〉〉（HgG/HgT）2000. A simple outline of Hg release in landfill could be drawn： with increasing of filling time, degradation undergoes different biodegradation, accordingly, gaseous mercury goes through small, more, and small proportion to total mercury. Distribution of Hg in plants was inhomogeneous, following the order of leaf〉root〉stem. The highest value of leaf may be associated with higher atmospheric Hg from landfill. Ligneous plants （e.g. Phyllostachys glanca, Prunus salicina and Ligustrum lucidum） are capable of enriching more Hg than herbaceous plants.     

Toxic effects of acetochlor, methamidophos and their combination on nifH gene in soil

Toxic effects of two agrochemicals on nifH gene in agricultural black soil were investigated using denaturing gradient gel electrophoresis （DGGE） and sequencing approaches in a microcosm experiment. Changes of soil nifH gene diversity and composition were examined following the application of acetochlor, methamidophos and their combination. Acetochlor reduced the nifH gene diversity （both in gene richness and diversity index values） and caused changes in the nifH gene composition. The effects of acetochlor on nifH gene were strengthened as the concentration of acetochlor increased. Cluster analysis of DGGE banding patterns showed that nifH gene composition which had been affected by low concentration of acetochlor （50 mg/kg） recovered firstly. Methamidophos reduced nifH gene richness that except at 4 weeks. The medium concentration of methamidophos （150 mg/kg） caused the most apparent changes in nifH gene diversity at the first week while the high concentration of methamidophos （250 mg/kg） produced prominent effects on nifH gene diversity in the following weeks. Cluster analysis showed that minimal changes of nifH gene composition were found at 1 week and maximal changes at 4 weeks. Toxic effects of acetochlor and methamidophos combination on nifH gene were also apparent. Different nifH genes （bands） responded differently to the impact of agrochemicals： four individual bands were eliminated by the application of the agrochemicals, five bands became predominant by the stimulation of the agrochemicals, and four bands showed strong resistance to the influence of the agrochemicals. Fifteen prominent bands were partially sequenced, yielding 15 different nifH sequences, which were used for phylogenetic reconstructions. All sequences were affiliated with the alpha- and beta-proteobacteria, showing higher similarity to eight different diazotrophic genera.     

Enzymatic oxidation of aqueous pentachlorophenol

The influence of the nonionic surfactant Tween 80 on pentachlorophenol （PCP） oxidation catalyzed by horseradish peroxidase was studied. The surfactant was tested at concentrations below and above its critical micelle concentration （CMC）. Enhancement of PCP removal was observed at sub-CMCs. The presence of Tween 80 in the reaction mixture reduced enzyme inactivation which occurred through a combination of free radical attack and sorption by precipitated products. A simple first-order model was able to simulate time profiles for enzyme inactivation in the presence or absence of Tween 80. At supra-CMCs, the surfactant caused noticeable reductions in PCP removal, presumably through micelle partitioning of PCP which precluded the hydrophobic PCP molecule from interacting with the enzyme.     

Decomposition of alachlor by ozonation and its mechanism

Decomposition and corresponding mechanism of alachlor, an endocrine disruptor in water by ozonation were investigated. Results showed that alachlor could not be completely mineralized by ozone alone. Many intermediates and final products were formed during the process, including aromatic compounds, aliphatic carboxylic acids, and inorganic ions. In evoluting these products, some of them with weak polarity were qualitatively identified by GC-MS. The information of inorganic ions suggested that the dechlorination was the first and the fastest step in the ozonation of alachlor.     

Arsenic uptake by arbuscular mycorrhizal maize （Zea mays L.） grown in an arsenic-contaminated soil with added phosphorus

The effects of arbuscular mycorrhizal (AM) fungus (Glomus mosseae) and phosphorus (P) addition (100 mg/kg soil) on arsenic (As) uptake by maize plants (Zea mays L.) from an As-contaminated soil were examined in a glasshouse experiment.Non-mycorrhizal and zero-P addition controls were included.Plant biomass and concentrations and uptake of As,P,and other nutrients,AM colonization,root lengths,and hyphal length densities were determined.The results indicated that addition of P significantly inhibited root colonization and development of extraradical mycelium.Root length and dry weight both increased markedly with mycorrhizal colonization under the zero-P treatments,but shoot and root biomass of AM plants was depressed by P application.AM fungal inoculation decreased shoot As concentrations when no P was added,and shoot and root As concentrations of AM plants increased 2.6 and 1.4 times with P addition,respectively.Shoot and root uptake of P,Mn,Cu,and Zn increased,but shoot Fe uptake decreased by 44.6%,with inoculation, when P was added.P addition reduced shoot P,Fe,Mn,Cu,and Zn uptake of AM plants,but increased root Fe and Mn uptake of the nonmycorrhizal ones.AM colonization therefore appeared to enhance plant tolerance to As in low P soil,and have some potential for the phytostabilization of As-contaminated soil,however,P application may introduce additional environmental risk by increasing soil As mobility.     

Degradation of metabolites accumulated of benzo[a]pyrene by coupling Penicillium Chrysogenum with KMnO4

Several main metabolites of benzo[a]pyrene （BaP） formed by Penicillium chrysogenum, Benzo[a]pyrene-1,6-quinone （BP 1,6- quinone）, trans-7,8-dihydroxy-7,8-dihydrobenzo[a]pyrene （BP 7,8-diol）, 3-hydroxybenzo[a]pyrene （3-OHBP）, were identified by high-performance liquid chromatography （HPLC）. The three metabolites were liable to be accumulated and were hardly further metabolized because of their toxicity to microorganisms. However, their further degradation was essential for the complete degradation of BaP. To enhance their degradation, two methods, degradation by coupling Penicillium chrysogenum with KMnO4 and degradation only by Penicillium chrysogenum, were compared; Meanwhile, the parameters of degradation in the superior method were optimized. The results showed that （1） the method of coupling Penicillium chrysogenum with KMnO4 was better and was the first method to be used in the degradation of BaP and its metabolites; （2） the metabolite, BP 1,6-quinone was the most liable to be accumulated in pure cultures; （3） the effect of degradation was the best when the concentration of KMnO4 in the cultures was 0.01% （w/v）, concentration of the three compounds was 5 mg/L and pH was 6.2. Based on the experimental results, a novel concept with regard to the bioremediation of BaP-contaminated environment was discussed, considering the influence on environmental toxicity of the accumulated metabolites.     

Impact of atrazine and nitrogen fertilizers on the sorption of chlorotoluron in soil and model sorbents

Sorption of chlorotoluron in ammonium sulfate, urea and atrazine multi-solutes system was investigated by batch experiments. The results showed application of nitrogen fertilizers to the soil could affect the behavior of chlorotoluron. At the same concentration of N, sorption of chlorotoluron decreased as the concentration of atrazine increased on the day 0 and 6 in soil, respectively. The sorption of chlorotoluron increased from 0 to 6 d when soils were preincubated with deionized water, ammonium sulfate and urea solution for 6 d. That indicated incubation time was one of the most important factors for the sorption of chlorotoluron in nitrogen fertilizers treatments. The individual sorption isotherms of chlorotoluron in rubbery polymer and silica were strictly linear in single solute system, but there were competition sorption between pesticides or between pesticides and nitrogen fertilizers. That indicated the sorption taken place by concurrent solid-phase dissolution mechanism and sorption on the interface of water-organic matter or water-mineral matter.