兽用抗生素磺胺二甲嘧啶对稻田N2O排放的影响及其微生物机制

为研究磺胺类兽用抗生素对稻田N_2O排放的影响及其微生物机制,采用田间原位观测试验,对比分析不同浓度磺胺二甲嘧啶(sulfamethazine,SMZ)对稻田N_2O排放及硝化反硝化过程底物和相关功能基因丰度的影响.本试验共设5个处理,分别为:无肥料无抗生素(CK);猪粪为基肥,尿素为追肥,分别添加0、5、15和30 mg·kg~(-1)的SMZ处理(SMZ0、SMZ5、SMZ15和SMZ30),在整个水稻生长季定期采集和分析土壤和气体样品.结果表明,不同浓度SMZ均未改变稻田N_2O排放的季节性规律,整个观测期N_2O排放通量,与SMZ0处理相比,SMZ15有显著差异(P 0. 05),SMZ30和SMZ5无显著差异(P 0. 05).中、高浓度处理SMZ15和SMZ30在均值水平上增加了N_2O累积排放量,分别是SMZ0处理的3. 47和4. 67倍,且增加了土壤NO_3~--N含量.与SMZ0处理相比,中、高浓度处理对土壤总细菌16S rRNA基因丰度、硝化过程中氨氧化古菌AOA amoA和氨氧化细菌AOB amoA基因丰度以及反硝化过程中的nirK、nirS和nosZ基因丰度均有明显的激活作用(P 0. 05),低浓度处理SMZ5对各基因丰度则有轻微抑制作用.具体表现为SMZ30、SMZ15与SMZ0处理的16S rRNA、AOA amoA、AOB amoA以及nirK、nirS、nosZ基因丰度比值的平均值分别为:1. 58、1. 77、2. 15、1. 38、1. 33、1. 42和1. 24、1. 37、1. 08、1. 65、1. 11、1. 64,而SMZ5与SMZ0处理的6个上述基因丰度比值均小于1,仅分别为0. 80、0. 99、0. 92、0. 76、0. 76和0. 77. N_2O排放通量与nir K基因丰度呈极显著正相关(P 0. 01),表明SMZ通过影响反硝化菌活性进而对N_2O排放产生作用.因此,兽用抗生素对农田的污染不可忽视,应从源头上合理控制使用,以减少其环境生态风险.     

典型油田多环芳烃污染对土壤反硝化微生物群落结构的影响

油田区土壤具有潜在的PAHs(polycyclic aromatic hydrocarbons)污染风险,而以硝酸根为电子受体的反硝化作用可能在PAHs的厌氧代谢中起到重要作用.以具有50多年历史的江汉油田区域为对象,从该油田的油井口附近采集了9个土壤样品,编号为JH-1~JH-9,以反硝化相关的nir K(Cu-亚硝酸还原酶基因)和nirS(细胞色素cd1-亚硝酸还原酶基因)为分子标识,通过定量PCR及克隆文库结合T-RFLP(terminal-restriction fragment length polymorphism)的方法,研究典型油田区土壤反硝化微生物的群落结构,并探讨其与土壤环境因子之间的关系.结果表明,该油田区土壤中nirK基因的丰度高于nirS基因,PAHs含量最高的土壤样品(JH-4)中反硝化功能基因nir K和nir S的丰度均最低,相关性分析表明,土壤nir K及nirS基因的丰度均与土壤PAHs含量呈显著负相关(nirK:R2=0.54,P0.05;nirS:R~2=0.58,P0.05).克隆文库及T-RFLP的结果则表明,该油田土壤中nirK基因的群落组成在不同样品间的变异较大,且PAHs含量最高的JH-4中该基因的群落组成与其它各样品有明显的不同,RDA(redundancy analysis)的分析结果进一步表明除有效氮、有效磷外,土壤PAHs含量也是影响nirK型反硝化微生物群落组成的重要因子.相较于nirK,该油田区土壤中nirS基因的群落组成在不同样品间的差异较小,但发现nirS型假单胞菌的丰度与土壤PAHs含量呈正相关,表明具备较强有机污染物降解能力的假单胞菌属可能在该区域土壤PAHs的反硝化代谢中起到重要作用.     

生物炭对华北农田土壤N2O通量及相关功能基因丰度的影响

为了探寻施用生物炭对农田土壤氧化亚氮(N_2O)的减排效果和机制,于2015年3月27日至6月5日,利用盆栽实验研究了施用生物炭(CK,C1:5%,C2:10%,C3:15%,C4:30%)(质量分数)对华北农田土壤N_2O通量、氨单加氧酶(amo A)、亚硝酸盐还原酶(nir S、nir K)以及氧化亚氮还原酶(nos Z)基因丰度的影响.结果表明:(1)施用低量生物炭(5%)能够促进N_2O排放,施用中、高量生物炭可以起到抑制N_2O排放的效果,且生物炭用量为15%时减排效果最佳.(2)实验初期,施用生物炭对土壤硝化反硝化基因丰度影响较大,AOA和nir S基因丰度与生物炭施用量呈极显著正相关关系,nir K基因丰度与生物炭施用量呈显著正相关关系,AOB和nos Z基因丰度与生物炭施用量呈显著负相关关系;实验末期,AOA丰度与生物炭施用量表现为显著负相关关系,AOB丰度与生物炭施用量表现为显著正相关关系.(3)实验初期,N_2O排放通量与AOA、nir S基因呈现极显著的负相关关系,说明在土壤含水量较高的条件下,N_2O的产生受AOA、nir S基因丰度控制调节;实验末期,N_2O排放通量与nos Z基因呈现极显著正相关关系,说明在土壤含水量较低的条件下,N_2O的产生受nos Z基因丰度控制调节.本研究结果表明施用生物炭能够增加硝化反硝化功能基因丰度,并降低N_2O的排放,为华北农田合理施用生物炭提供了一定的理论依据.     

磷差异性调控水稻根际nirK/nirS型反硝化菌组成与丰度

磷作为一种重要的生命元素,对反硝化微生物的活性和功能有重要影响.反硝化功能基因nir K和nir S是编码亚硝酸还原酶的两种同工酶基因,但是,磷对具有同种功能的nir K和nir S型反硝化菌的调控是否存在差异尚不十分清楚.本文采集严重缺磷红壤性水稻土设置水稻盆栽试验,通过外源添加磷肥设置对照(CK,P:0 mg·kg-1),低磷(P1,P:15 mg·kg-1),高磷(P2,P:30 mg·kg-1)这3个磷添加水平,研究不同磷水平对水稻亚硝酸还原酶基因的组成与丰度的调控作用.结果表明,在长期缺磷土壤上施用磷肥对含亚硝酸还原酶基因nir K和nir S的细菌种群的调控作用有明显差异.不管是根际还是非根际土,含nir S的微生物种群均对施磷水平表现敏感,尤其是高磷(P2)水平,施磷可导致nir S丰度提高2～3倍,同时显著改变nir S型反硝化微生物的组成结构.相比之下,施磷后含nir K基因的微生物组成结构和丰度变异较小.与非根际土壤相比,高磷水平条件下根际土中nir S的基因拷贝数和群落结构均发生了显著变化,缺磷和低磷条件下水稻生长只引起根际土nir S种群组成结构发生显著变化,但其丰度与非根际无显著差异.但不同磷水平条件下nir K的基因丰度和组成结构在根际和非根际土之间几乎无显著变化.综上所述,在严重缺磷水稻土中加施磷肥会显著提高水稻根际和非根际土中nir K和nir S型反硝化菌数量,并改变其种群组成结构,且nir S比nir K型种群响应更敏感.不同磷水平条件下的水稻根系生长均显著改变了根际土壤中nir K和nir S种群组成结构,但除了在高磷水平条件下显著增加了nir S丰度外,对nir K和nir S丰度均影响较小.研究结果可为进一步深入探究施肥对土壤反硝化过程的影响提供理论依据.     

长期施肥对稻田不同土层反硝化细菌丰度的影响

为了探讨长期施肥对稻田不同土层关键反硝化功能种群丰度的影响及核心驱动因子,以湖南宁乡长期施肥定位试验田为平台,选取不施肥（CK）、全量化肥（NPK）和秸秆还田（ST）3个处理,结合实时荧光定量PCR（qPCR）技术,系统分析了稻田不同土层（0~10,10~20,20~30,30~40cm）关键反硝化功能基因（narG、nirK和nirS）的丰度及其与土壤理化性质的内在联系.结果表明,相比于不施肥处理（CK）,施肥处理（NPK和ST）在0~40cm土层土壤SOC、TN、NO₃^--N、NH₄⁺-N和Olsen-P分别显著增加了2.2%~83.6%,3.5%~58.3%,70.8%~222.1%,0.9%~83.7%和16.5%~94.5%,pH值下降了0.31~0.67个单位;长期施用化肥和秸秆使narG、nirK和nirS基因丰度分别增加0.75~7.18倍,1.57~3.02倍和0.53~3.81倍,其中秸秆还田对反硝化细菌数量的影响比单施化肥更显著;稻田narG、nirK和nirS反硝化型细菌的丰度随土层深度增加而逐渐降低,具有明显的垂直分布特征;RDA分析结果显示,土壤养分如SOC和TN是影响水稻土narG、nirK和nirS反硝化型细菌垂直分布的关键因子,而pH值是调控反硝化细菌在稻田底土分布的核心驱动因子.研究结果可为提升稻田土壤肥力和减少稻田氮素损失和温室气体排放提供理论依据.     

CO2倍增和施氮对水稻不同生长期土壤反硝化细菌丰度的影响

随着全球气候变化的不断加剧,大气CO₂浓度呈明显增加趋势,这将间接影响土壤-植物-微生物系统的氮循环过程.为研究典型水稻土壤反硝化细菌对CO₂浓度升高的响应规律和机制,借助水稻密闭培养箱,运用实时荧光定量聚合酶链式反应（Real-Time qPCR）分子技术,设置不施氮（0 mg/kg）和常规施氮（100 mg/kg）2个处理,研究CO₂倍增对水稻不同生长期土壤关键反硝化功能细菌（narG、nirK和nirS型）丰度的影响.结果表明:①在2种施氮水平,CO₂倍增显著促进了水稻分蘖期、孕穗期、扬花期和成熟期水稻根系生长（增幅为2.96%~28.4%）、地上部生物量增加（增幅为7.1%~107.3%）以及成熟期籽粒干质量的增加（增幅为19.5%和38.0%）,具有显著的增产效应.②反硝化细菌丰度对CO₂倍增的响应与生育期及施氮水平有关,CO₂倍增在2个施氮水平均抑制分蘖期反硝化细菌的繁殖,显著增加孕穗期反硝化细菌数量;在水稻扬花期,CO₂倍增促进了施氮处理narG和nirS型反硝化细菌数量的增加,在成熟期抑制未施氮处理下narG、nirK和nirS型反硝化细菌的生长.另外,narG、nirK、nirS型反硝化细菌丰度整体表现为narG > nirS > nirK,且随水稻的生长,其在成熟期的丰度均呈降低趋势.nirK和nirS基因同属亚硝酸还原酶,但nirS基因丰度高于nirK,且对CO₂倍增和施氮的响应有所差异.研究显示,CO₂倍增可显著增加水稻生长和产量,不同施氮水平对稻田土壤反硝化细菌丰度的影响存在差异.      

咸水滴灌对棉田土壤N2O排放和反硝化细菌群落结构的影响

淡水资源短缺是干旱区农业可持续发展所面临的严峻问题,合理利用咸水灌溉是缓解淡水资源不足的重要手段.长期咸水灌溉会导致土壤盐分积累,进而影响氮素的转化和N_2O的排放.本研究通过10 a咸水灌溉试验,探究咸水灌溉对棉田土壤N_2O排放、反硝化细菌丰度和群落结构组成的影响.试验采用灌溉水盐度和施氮量两因子2×2随机区组设计,其中灌溉水盐度(以电导率表示)设置2个水平:0.35 dS·m~(-1)和8.04 dS·m~(-1),施氮量设2个水平:0 kg·hm~(-2)和360 kg·hm~(-2)(分别用SFN0、SHN0、SFN360和SHN360表示).结果表明,长期咸水滴灌棉田土壤盐分、含水量和NH~+_4-N含量显著增加,pH值、NO~-_3-N、有机质和全氮含量显著降低.咸水灌溉处理显著抑制N_2O排放,不施氮肥和施氮肥处理下分别较淡水灌溉降低45.19%和43.50%.氮肥施用显著增加N_2O排放,施肥处理N_2O排放较不施肥处理增加161%.不施肥条件下,咸水灌溉显著降低反硝化酶活性、nirK、nirS和nosZ基因丰度,α多样性.施肥条件下,咸水灌溉对nosZ型反硝化细菌的丰度无显著影响,但显著降低反硝化酶活性和nirK、nirS基因丰度.咸水灌溉和氮肥施用共同改变nirK、nirS和nosZ型反硝化细菌群落结构,灌溉水盐度对于反硝化细菌群落结构的影响要大于施肥.Lefse分析显示nirK、nirS和nsoZ型反硝化细菌差异物种随着灌溉水盐度的增加而增加,咸水灌溉显著改变反硝化细菌群落结构,导致优势种群数量增加.上述结果表明,长期咸水灌溉降低土壤N_2O排放,但会导致土壤盐分的持续上升,nosZ、nirK和nirS丰度的增加会促进N_2O排放.     

生物炭和有机肥对菜地土壤N2O排放及硝化、反硝化微生物功能基因丰度的影响

通过室内培养试验和实时荧光定量PCR技术,研究了田间施用生物炭和有机肥对菜地土壤氧化亚氮(N_2O)排放、氨单加氧酶(amo A)和亚硝酸盐还原酶(nir S、nir K)、氧化亚氮还原酶(nos Z)基因丰度的影响,并探讨功能基因丰度对N_2O排放的影响.试验设置5个处理:CK(对照)、N(尿素)、N+BC(尿素和生物炭)、N+M(尿素和有机肥)和N+BC+M(尿素、生物炭和有机肥).结果表明,与CK处理相比,各施肥处理均降低了土壤氨氧化细菌(AOB)和氨氧化古菌(AOA)丰度,增加了nir K、nir S和nos Z基因丰度,并提高了培养期间N_2O累积排放量.与N处理相比,N+BC处理的土壤p H值提高了11.1%,并增加了AOB、AOA、nir S、nir K和nos Z基因丰度,增幅分别为105.8%、57.3%、22.0%、176.2%和204.9%,同时显著降低了培养期间N_2O累积排放量,降幅为58.1%;N+M处理增加了nir K和nir S基因丰度,增幅分别为58.8%和7.1%,对N_2O排放的影响不显著;N+BC+M处理增加了AOB、nir K、nir S和nos Z基因丰度,增幅分别为30.7%、68.7%、6.5%和84.5%,降低了N_2O累积排放量,降幅为14.4%.生物炭通过增加amo A、nir S和nir K基因丰度间接增加N_2O排放,同时通过增加nos Z基因丰度促进N_2O还原,综合效应表现为降低了菜地土壤N_2O排放.因此,通过施用生物炭改善土壤性质,增加功能基因丰度,降低土壤N_2O排放,是一种较好的N_2O减排措施.施用有机肥可以增加反硝化作用功能基因丰度,但对N_2O减排效果不显著.     

稻田土壤不同水分条件下硝化/反硝化作用及其功能微生物的变化特征

以湖南桃源县一长期种植水稻的酸性土壤为研究对象,在微宇宙培养条件下设置了4个水分梯度处理,分别为田间持水量(water holding capacity,WHC)的30%、60%、90%和淹水2 cm深.考察了水分条件变化对硝化和反硝化作用影响,并结合定量PCR和限制性末端片段长度多态性(T-RFLP)技术研究了硝化-反硝化微生物的响应特征.结果表明,30%WHC处理土壤无明显的硝化和反硝化作用发生,硝化作用主要发生于60%WHC和90%WHC处理土壤,90%WHC处理土壤硝化作用明显强于60%WHC,并检测到明显的N2O释放,表明该水分条件可能发生了硝化-反硝化耦合作用.淹水处理土壤氧化还原势Eh显著低于非淹水处理土壤,无明显的硝化作用发生,但能检测到N2O释放且释放量小于90%WHC处理土壤.除培养初期(7 d)外,反硝化功能基因nirS和nirK,以及氨氧化细菌(AOB)amoA基因的丰度先随着水分增加而增加,并在淹水处理中小幅下降,三者之间呈明显的正相关关系,且AOB amoA、nirS和nirK基因丰度均在90%WHC处理中最高,与该处理中硝化和反硝化活性最高相一致.T-RFLP结果表明,培养2周后,nirS基因为代表的反硝化微生物群落组成对水分梯度变化产生明显响应,Eh和含水率Cw是影响其群落组成的主要因子.     

生物质炭对双季稻田土壤反硝化功能微生物的影响

目前,基于田间条件下生物质炭添加对稻田反硝化微生物的调控效应还不甚明确.为此,本研究采用小区试验,通过在双季稻田添加不同量的小麦秸秆生物质炭(0、24和48 t·hm-2,分别用CK、LC和HC代表),结合实时荧光定量PCR(q PCR)和末端限制性片段长度多态性(T-RFLP)分析技术,研究了生物质炭添加对双季稻田休闲季和水稻季土壤反硝化微生物相关功能基因(调控硝酸还原酶的nar G基因,亚硝酸还原酶的nir K基因和氧化亚氮还原酶的nos Z基因)的影响.由于生物质炭呈碱性,添加到土壤后,可提高稻田休闲季土壤p H 0. 2～0. 8个单位.生物质炭本身含有部分可溶性N,因此,添加生物质炭可增加休闲季土壤铵态氮(NH_4~+-N)和硝态氮(NO_3~--N)含量,增幅分别达21. 1%～32. 5%和63. 0%～176. 0%,但由于其吸附作用,降低了水稻季NH_4~+-N含量48. 8%～60. 1%.生物质炭添加增加了休闲季微生物生物量氮(MBN)含量,这可能是由于生物质炭较大的比表面积为微生物生存提供了适宜的环境,可利用养分的增加促进了微生物的生长.与对照相比,休闲季生物质炭引起的NH_4~+-N和NO_3~--N含量增加,促进NH_4~+-N向NO_3~--N的转化,进而增加nar G和nos Z的基因丰度(P0. 05),同时,生物质炭处理p H的提高促进了nos Z的基因丰度的增加,显著改变了反硝化功能基因nar G和nos Z的群落结构,并以此对反硝化作用产生影响,但未对休闲季氧化亚氮(N_2O)排放产生影响.而在水稻季,生物质炭增加了土壤nos Z的基因丰度(P 0. 05),HC处理增加了nir K基因丰度(P 0. 05),这也是导致水稻季HC处理N_2O排放增加的重要原因.生物质炭通过降低水稻季土壤NH_4~+-N含量,改变了nir K和nos Z基因的群落结构,而nar G基因群落结构的变化影响了土壤N_2O排放.综上所述,生物质炭可通过改变双季稻田土壤性质,来影响参与土壤反硝化作用的相关微生物,进而影响土壤N_2O排放及NO_3~--N的淋失.     

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.     

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.     

Potential of plant polyphenol oxidases in the decolorization and removal of textile and non-textile dyes

In this study an effort has been made to use plant polyphenol oxidases; potato （Solanum tuberosum） and brinjal （Solanum melongena）, for the treatment of various important dyes used in textile and other industries. The ammonium sulphate fractionated enzyme preparations were used to treat a number of dyes under various experimental conditions. Majority of the treated dyes were maximally decolorized at pH 3.0. Some of the dyes were quickly decolorized whereas others were marginally decolorized. The initial first hour was sufficient for the maximum decolorization of dyes. The rate of decolorization was quite slow on long treatment of dyes. Enhancement in the dye decolorization was noticed on increasing the concentration of enzymes. The complex mixtures of dyes were treated with both preparations of polyphenol oxidases in the buffers of varying pH values. Potato polyphenol oxidase was significantly more effective in decolorizing the dyes to higher extent as compared to the enzyme obtained from brinjal polyphenol oxidase. Decolorization of dyes and their mixtures, followed by the formation of an insoluble precipitate, which could be easily removed simply by centrifugation.     

Removal of heavy metals from sewage sludge by low costing chemical method and recycling in agriculture

ＲｅｍｏｖａｌｏｆｈｅａｖｙｍｅｔａｌｓｆｒｏｍｓｅｗａｇｅｓｌｕｄｇｅｂｙｌｏｗｃｏｓｔｉｎｇｃｈｅｍｉｃａｌｍｅｔｈｏｄａｎｄｒｅｃｙｃｌｉｎｇｉｎａｇｒｉｃｕｌｔｕｒｅＷｕＱｉｔａｎｇ，ＮｙｉｒａｎｄｅｇｅＰａｓｃａｓｉｅ，ＭｏＣｅｈｕｉＦ...     

Isolation and preliminary characterization of a 3-chlorobenzoate degrading bacteria

A study was conducted to compare the diversity of 2-, 3-, and 4-chlorobenzoate degraders in two pristine soils and one contaminated sewage sludge. These samples contained strikingly different populations of mono-chlorobenzoate degraders. Although fewer cultures were isolated in the uncontaminated soils than contaminated one, the ability of microbial populations to mineralize chlorobenzoate was widespread. The 3- and 4-chlorobenzoate degraders were more diverse than the 2-chlorobenzoate degraders. One of the strains isolated from the sewage sludge was obtained. Based on its phenotype, chemotaxonomic properties and 16S rRNA gene, the organism S-7 was classified as Rhodococcus erythropolis. The strain can grow at temperature from 4 to 37℃. It can utilize several （halo）aromatic compounds. Moreover, strain S-7 can grow and use 3-chlorobenzoate as sole carbon source in a temperatures range of 10-30℃ with stoichiometric release of chloride ions. The psychrotolerant ability was significant for bioremediation in low temperature regions. Catechol and chlorocatechol 1,2-dioxygenase activities were present in cell free extracts of the strain, but no （chloro）catechol 2,3- dioxygenase activities was detected. Spectral conversion assays with extracts from R. erythropolis S-7 showed accumulation of a compound with a similar UV spectrum as chloro-cis,cis-muconate from 3-chlorobenzoate. On the basis of these results, we proposed that S-7 degraded 3-chlorobenzoate through the modified ortho-cleave pathway.     

Single and joint effects of pesticides and mercury on soil urease

The influence of two pesticides including chlorimuron-ethyl and furadan and mercury （Hg） on urease activity in 4 soils （meadow burozem and phaeozem） was investigated. The soils were exposed to various concentrations of the two pesticides and Hg individually and simultaneously. Results showed that there was a close relationship between urease activity and organic matter content in soil. Chlorimuron-ethyl and furadan could both activate urease in the 4 soils. The maximum increment of urease activity by chlorimuronethyl was up to 14%-18%. There was almost an equal increase （up to 13%-21%） in the urease activity by furadan. On the contrary, Hg markedly inhibited soil urease activity. A logarithmic equation was used to describe the relationship （P〈0.05） between the concentration of Hg and the activity of soil urease in the 4 tested soils. Semi-effect dose （ED50） values by the stress of Hg based on the inhibition of soil urease in the 4 soils were 88, 5.5, 24 and 20 mg/kg, respectively, according to the calculation of the corresponding equations. The interactive effect of chlorimuron-ethyl or furadan with metal Hg on soil urease was mainly synergic at the highest tested concentrations.     

Organochlorine pesticides in soils under different land usage in the Taihu Lake region, China

A field study was conducted in the Taihu Lake region, China in 2004 to reveal the organochlorine pesticide concentrations in soils after the ban of these substances in the year 1983. Thirteen organochlorine pesticides (OCPs) were analyzed in soils from paddy field, tree land and fallow land. Total organochlorine pesticide residues were higher in agricultural soils than in uncultivated fallow land soils. Among all the pesticides, ΣDDX (DDD, DDE and DDT) had the highest concentration for all the soil samples, ranging from 3.10 ng/g to 166.55 ng/g with a mean value of 57.04 ng/g and followed by ΣHCH, ranging from 0.73 ng/g to 60.97 ng/g with a mean value of 24.06 ng/g. Dieldrin, endrin, HCB and α-endosulfan were also found in soils with less than 15 ng/g. Ratios of p,p'-(DDD DDE)/DDT in soils under three land usages were: paddy field ＞ tree land ＞ fallow land, indicating that land usage inlfuenced the degradation of DDT in soils. Ratios of p,p'-(DDD DDE)/DDT ＞1, showing aged residues of DDTs in soils of the Taihu Lake region. The results were discussed with data from a former study that showed very low actual concentrations of HCH and DDT in soils in the Taihu Lake region, but according to the chemical half-lives and their concentrations in soils in 1980s, the concentration of DDT in soils seemed to be underestimated. In any case our data show that the ban on the use of HCH and DDT resulted in a tremendous reduction of these pesticide residues in soils, but there are still high amounts of pesticide residues in soils, which need more remediation processes.     

Alterations of organ histopathology and metallolhionein mRNA expression in silver barb, Puntius gonionotus during subchronic cadmium exposure

Common silver barb,Puntius gonionotus,exposed to the nominal concentration of 0.06 mg/L Cd for 60 d,were assessed for histopathological alterations(gills,liver and kidney),metal accumulation,and metallothionein(MT)mRNA expression.Fish exhibited pathological symptoms such as hypertrophy and hyperplasia of primary and secondary gill lamellae,vacuolization in hepatocytes,and prominent tubular and glomerular damage in the kidney.In addition,kidney accumulated the highest content of cadmium,more than gills and liver.Expression of MT mRNA was increased in both liver and kidney of treated fish.Hepatic MT levels remained high after fish were removed to Cd-free water.In contrast,MT expression in kidney was peaked after 28 d of treatment and drastically dropped when fish were removed to Cd-free water.The high concentrations of Cd in hepatic tissues indicated an accumulation site or permanent damage on this tissue.