富硒蚯蚓的培养及其硒富集作用研究

以赤子爱胜蚓为生物载体,在蚯蚓饲料中添加亚硒酸钠进行富硒蚯蚓培养试验,利用蚯蚓的生物富集作用将无机硒转化为有机硒。以存活率、硒含量为优化指标,研究5种硒浓度(20、40、60、80、100 mg·kg-1)和4种培养时间(第15、30、45、60 d)对蚯蚓生长和硒含量的影响规律,筛选存活率高、硒含量高、培养时间短的富硒蚯蚓培养方法。试验结果表明:蚯蚓饲料中亚硒酸钠含量越高、培养时间越长,蚯蚓的存活率越低,硒含量越高,且当硒添加量为80 mg·kg-1,培养时间为45 d时,培养条件最优,此时蚯蚓体内硒含量可达33.25 mg·kg-1。     

莫能菌素对蚯蚓的生态毒理效应

采用人工土壤法测定了不同浓度莫能菌素对蚯蚓的急性毒性和对蚯蚓生长、生存、繁殖以及超氧化物歧化酶（SOD）、谷胱甘肽-硫转移酶（GSH-S）、腺三磷酸酶（ATPase）、乙酰胆碱酯酶（T-CHE）活性的影响。结果表明,莫能菌素对蚯蚓14d的LC50为75.883mg·kg-1;当莫能菌素含量达到50mg·kg-1时显著影响蚯蚓的生存（P〈0.05）,死亡率达到20%;当莫能菌素含量达到25mg·kg-1时显著影响蚯蚓的生长（P〈0.05）;药物的染毒浓度与对蚯蚓生存和繁殖的抑制存在明显的剂量-效应关系。染毒21d后,低浓度的兽药莫能菌素就能显著影响蚯蚓体内SOD、GSH-S、ATPase、T-CHE的活性,药物浓度和酶活性间具有明显的剂量-效应关系。莫能菌素对土壤动物蚯蚓显示有毒性作用,对土壤环境具有潜在的生态风险。     

菊糖对蚯蚓粪肥性状的影响及调控过程

通过蚯蚓堆肥试验,探讨菊糖对蚯蚓的益生作用及高活力蚯蚓粪肥的形成过程,可为滩涂生态农牧化模式构建及障碍土壤修复提供理论依据.以牛粪、赤子爱胜蚓(Eisenia fetida)和菊糖为材料,设置纯牛粪养殖蚯蚓(EM)和添加菊糖的牛粪养殖蚯蚓(IEM)处理,养殖周期为40 d,每隔10 d采样分析,研究了牛粪–蚯蚓肠道内容...     

铜和草甘膦对蚯蚓的毒性效应研究

污染物的交互作用及其生态毒理效应已成为环境科学研究的主要方向。含铜杀菌剂与除草剂草甘膦[N-(膦酸甲基)甘氨酸]是农业生产中常见的农药,其在土壤中共存可能造成复合污染。草甘膦中含有羧基、氨基、磷酸基等配位基团对重金属阳离子和有机阳离子有很强的络合能力,草甘膦可以影响重金属的生物有效性、毒性以及重金属在生物体内的积累。本文以赤子爱胜蚓(Eisenia fetida)为例,采用室内模拟试验,单一试验设置5个铜浓度25 mg·kg-1、50 mg·kg-1、100 mg·kg-1、200 mg·kg-1和400 mg·kg-1,5个草甘膦浓度25mg·kg-1、50 mg·kg-1、100 mg·kg-1、200 mg·kg-1和500 mg·kg-1,并以去离子水做空白对照;复合试验设置2个铜浓度(低铜浓度25 mg·kg-1和高铜浓度200 mg·kg-1)分别与5个草甘膦浓度(0、25 mg·kg-1、50 mg·kg-1、100 mg·kg-1和200 mg·kg-1)复合;每个处理3次重复,试验期为35 d。研究了铜和草甘膦的单一及复合施用对蚯蚓的亚急性毒性效应。单一毒性试验结果表明,蚯蚓对低浓度铜的响应不敏感,而较高浓度铜使蚯蚓生物量的相对增长率受到显著抑制(α=0.01,r2=0.570),且未出现死亡,并且随着铜浓度的增加蚯蚓体内铜含量显著增加(α=0.01,r2=0.905)。与对照相比草甘膦的添加对蚯蚓的生长无显著影响。复合毒性试验结果表明,草甘膦和铜复合可以降低蚯蚓对铜的吸收,特别是当铜浓度较高时即200 mg·kg-1,草甘膦和铜复合可显著降低蚯蚓对铜的吸收,并且草甘膦能够减轻铜对蚯蚓生物量相对增长率的抑制,但差异并不显著。以上结果表明,铜比草甘膦的毒性大很多,铜与草甘膦复合能够减少蚯蚓对铜的吸收,缓解铜的毒性。因此本研究认为,在草甘膦和重金属共存的污染土壤中,草甘膦能够控制重金属的生物有效性和毒性。     

氯丹和灭蚁灵污染场地土壤对蚯蚓的毒性效应研究

以赤子爱胜蚓（Eisenia fetida）为供试生物,氯丹和灭蚁灵污染场地土壤为供试土壤,以蚯蚓体重及体内蛋白质含量、超氧化物歧化酶（SOD）和过氧化氢酶（CAT）活性为指标,进行了不同暴露时间（1、3、7、14 d）下场地土壤中氯丹和灭蚁灵污染胁迫对蚯蚓的毒性效应研究。结果表明,随着暴露时间的延长,蚯蚓体重在氯丹和灭蚁灵作用下受到显著抑制,蚯蚓体内蛋白质、SOD和CAT活性对氯丹和灭蚁灵响应不同,其敏感性大小为CAT〉SOD〉蛋白质;在一定暴露时间内,根据暴露-剂量效应关系,表明氯丹浓度为14.13 mg.kg-1、灭蚁灵浓度为4.14 mg.kg-1可能是使蚯蚓CAT活性达到最大值的临界浓度,同时也是SOD活性受到抑制的临界浓度,超过该临界浓度可能对蚯蚓产生生态毒性效应,这为场地风险评价和修复提供了基础数据。     

不同类型钾肥对滴灌甜菜产量及钾肥利用的影响

甜菜是我国及世界主要糖料作物之一,但钾肥的不合理利用严重制约了甜菜生产。以不同甜菜品种为研究对象,探究施用不同钾肥类型及不同钾肥用量对土壤钾素迁移、甜菜块根产量、产糖量及钾肥利用率的影响,以期确定适宜甜菜的钾肥类型及施用量。试验设置2个甜菜品种(KWS9899、KWS7748)、2种钾肥类型(氯化钾和硫酸钾)、4个钾肥施用量(以K2O计0、105、210、315 kg/hm²)。结果表明:(1)在距离滴灌带水平10、20 cm处,两种甜菜均以施用钾肥(K2O) 315 kg/hm²时土壤的速效钾迁移量最大,并且氯化钾肥的速效钾迁移量大于硫酸钾肥。同时,两种肥料类型下土壤中的速效钾均在20～40 cm土层中积累。(2)钾肥会提高甜菜块根产量和产糖量,以施用硫酸钾肥(K2O 210 kg/hm²)的产量与产糖量最高,且两种甜菜钾肥利用效率均随氯化钾肥和硫酸钾肥施用量的增加而下降。综上,施用钾肥可以增加耕层土壤速效钾和全钾含量,进而提高了甜菜块根产量和产糖量,增加了钾肥农学利用率。硫酸钾肥(K2O 210 kg/hm²)对甜菜的块根产量和产糖量增加效果最佳。     

蚯蚓（Eiseniafetida）细胞色素P450及抗氧化酶系对环境浓度苯并（a）芘的响应

以赤子爱胜蚓（Eiseniafetida）为供试生物,草甸棕壤为供试土壤,以蚯蚓微粒体细胞色素P450含量、抗氧化酶系以及谷胱甘肽转移酶活性为指标,进行了土壤中苯并（a）芘[B（a）P]暴露与酶活性的量-效关系研究。结果表明,在接近沈抚灌区实地污染状况B（a）P（0．1～2．0mg·kg-1）暴露下,第1、3、7以及第14d取样时,P450和SOD有较好的响应：SOD在第1、3d显著升高,而在第7、14d时降低;P450总体表现为低浓度B（a）P诱导下降低,高浓度下升高的趋势。CAT、POD以及GST的敏感性相对较差。比照其他4个指标,蚯蚓P450的敏感性更为优异,具有较好的应用前景。指标敏感性总体表现为：P450〉SOD〉CAT／POD〉GST。     

莫能菌素对蚯蚓（Eisenia fetida）活体DNA的损伤研究

通过单细胞凝胶电泳实验研究了不同浓度莫能菌素暴露对赤子爱胜蚓（Eisenia fetida）体腔细胞DNA的损伤,结果显示,50mg.kg-1莫能菌素处理组尾部DNA含量值最大、尾长值最大、Olive尾矩值最大,分别为34.539%、107.736μm和29.354;随着莫能菌素暴露剂量的增加,尾部DNA含量、Olive尾矩和尾长损伤频率增加;尾部DNA含量对莫能菌素暴露最为敏感,对照组和各处理组的尾部DNA含量之间均存在显著性差异（P〈0.05）;对照组与15、25、50 mg.kg-1处理组的Olive尾矩和尾长损伤频率之间均存在显著性差异（P〈0.05）;暴露浓度与尾部DNA含量、Olive尾矩和尾长具有良好的剂量-效应关系（P〈0.05）。实验结果表明,蚯蚓体腔细胞DNA损伤可作为指示莫能菌素影响的生物标志物,彗星试验是检测莫能菌素暴露对赤子爱胜蚓活体基因损伤的有效手段。     

蚯蚓在植物修复菲污染土壤中的作用

采用盆栽试验法,研究了蚯蚓（Pheretima sp.）在植物修复菲污染土壤中的作用。结果显示,试验浓度（20.05-322.06 mg.kg-1）范围内,蚯蚓活动促进了菲污染土壤中修复植物黑麦草（Lolium multiforum）的生长,其根冠比明显增大。添加蚯蚓72 d后,种植黑麦草的土壤中菲的去除率高达64.35%-93.40%,其平均去除率（80.92%）比无蚯蚓活动的土壤-植物系统（71.57%）提高9.35%,比无植物对照组（22.58%）提高57.34%。各种生物、非生物修复因子中,植物-微生物交互作用对菲去除的平均贡献率（53.56%）最为突出,比无蚯蚓活动时（47.48%）提高6.08%。说明蚯蚓活动可强化土壤-植物系统对土壤菲污染的修复作用。     

蚯蚓对天竺葵-土壤体系中Cd转运的影响研究

利用石墨炉(GF-990)原子吸收法对蚯蚓诱导下天竺葵-土壤体系中Cd的转移规律进行了研究。研究结果表明:在相同Cd浓度下,随着蚯蚓数量的增加,天竺葵各部位中Cd含量均有增高的趋势,10条蚯蚓对天竺葵-土壤体系中Cd转运的影响最大;在高Cd浓度(150 mg.kg-1)下,随着蚯蚓数量的增加(10~50条),天竺葵地上部分Cd的富集量明显大于地下部分,并且土壤中Cd含量明显出现降低的趋势,说明蚯蚓可以促进土壤Cd向天竺葵植物体中的转移。     

蚯蚓对电刺激的响应及其与蚯蚓粪胁迫分离的参数量化

蚯蚓与蚯蚓粪快速高效分离是规模化蚯蚓生物消解农业废弃物的关键技术环节。针对目前电刺激分离蚯蚓方法在实际生产应用中缺乏相关定量参数等问题，该研究探讨了蚯蚓分离率对电刺激方式、强度、通断电循环周期时长、通电时长及电刺激耦合物料含水率、物料厚度等因子的响应关系，并分析了电刺激对蚯蚓生理生化指标的影响。结果表明，电刺激可促进蚯蚓与蚓粪分离，间歇式电刺激方式较持续式电刺激具有更好的分离效果；间歇式电刺激方式下，蚯蚓分离率与电刺激强度、通断电循环周期时长、通电时长均呈显著的开口向下一元二次抛物线关系，蚯蚓生长的物料含水率适宜值为52.60%，物料厚度越小，越有利于电刺激分离率的提高；电刺激处理对蚯蚓体内过氧化氢酶、超氧化物歧化酶、乙酰胆碱酯酶、纤维素酶、可溶性蛋白含量无显著影响，但显著提高了丙二醛含量，蚯蚓经过5 d的恢复性驯养后，丙二醛含量可恢复至对照处理水平；综合蚯蚓分离率、技术参数易操作性等，采用间歇式电刺激方式分离蚯蚓，设计电刺激强度2 mA、通断电周期时长30 s、通电时长1 s，控制物料含水率50%左右、物料厚度20～25 cm，辅助210 lx白光光照，电刺激作业时间30 min，蚯蚓分离效率可达85%左右。研究可为快速高效分离蚯蚓活体与蚯蚓粪的设备研发与应用提供参考。     

赤子爱胜蚓Eisenia foetida体表形貌及润湿性的研究

以东北蚯蚓常见种赤子爱胜蚓（Eisenia foetida）为实验样本,对其处于收缩、静息和舒张三种状态下的体前部及体中部的体表形貌进行观察拍照和三维扫描,并对其体表接触角进行测量。结果表明收缩态、静息态和舒张态具有不同的非光滑单元密度及表面粗糙度,其接触角也存在显著性差别。收缩态非光滑单元密度和表面粗糙度最大,接触角也最大,即润湿性最差;舒张态非光滑单元密度和表面粗糙度最小,接触角同样最小,即润湿性最好;静息态各参数则处于中间水平。据此,初步建立了蚯蚓体表非光滑单元密度及表面粗糙度与表面润湿性之间的关系,进一步证实了波纹形非光滑结构模型所提出的波形比与润湿性之间的反比关系。     

Behavior and respiration responses of the earthworm Eisenia fetida to soil arsenite pollution

Soil arsenic (As) pollution from mining and industrial sources is a serious issue in China. Earthworms are considered ecosystem engineers and contribute to soil fertility development and maintenance of soil physico-chemical properties. In this study, earthworms were exposed to soils with different sodium arsenite concentrations (0, 5, 20, and 80 mg As kg^-1) for 60 d to investigate the changes in soil properties and the responses of the earthworms (e.g., burrowing activity and respiration). Earthworm burrowing activity decreased with increasing arsenite concentrations, and earthworm respiration was significantly lower in soils with 20 and 80 mg As kg^-1 compared to 0 mg As kg^-1. Changes in soil properties were also observed after incubation of As-amended soil with earthworms. Specifically, soil pH decreased, while soil electrical conductivity and contents of soil NH₃^--N, Olsen-P, and available K increased. Our results suggest that arsenite negatively impacts the metabolic activity of earthworms, leading to reduced burrowing activity, which in turn modifies the effects of earthworms on soil fertility and remediation.     

蚯蚓磨浆发酵产物减轻苹果连作障碍的机制

  【目的】  探究蚯蚓磨浆发酵产物防控苹果连作障碍的可行性。  【方法】  蚯蚓 (Eisenia foetida) 用湿法磨碎后,加入枯草芽孢杆菌 (Bacillus subtilis) 和黑曲霉菌 (Aspergillus niger) 发酵,发酵物用于试验。盆栽试验土壤采自27年龄的苹果园,供试作物为平邑甜茶幼苗。蚯蚓发酵物施入土壤,在苹果幼苗生长4个月后,调查苹果幼苗的生长状况,分析土壤中总微生物量、致病成分含量和有害真菌数量。  【结果】  施用蚯蚓磨浆发酵产物有效提高了苹果连作土壤中的细菌、放线菌数量,显著降低了真菌数量。与连作土对照相比,施用蚯蚓磨浆发酵产物处理连作土壤尖孢镰孢菌、层出镰孢菌、腐皮镰孢菌和串珠镰孢菌这4种有害真菌分别降低了65.6%、13.9%、84.2%和24.1%;有效降低了连作土壤中酚酸类物质的含量,其中根皮苷、根皮素和对羟基苯甲酸含量分别降低了74.2%、48.9%和67.0%;显著促进了植株根系的生长,提高了根系呼吸速率和抗氧化酶活性,根系呼吸速率、SOD、POD和CAT活性分别提高了69.4%、81.2%、81.1%和161.5%。蚯蚓磨浆发酵产物的添加极大促进了平邑甜茶幼苗植株的生长,植株干重是连作对照CK的2.58倍;同时提高了植株叶片的光合效率及叶绿素含量,其中光合效率比对照提高了46.5%,叶片叶绿素a和叶绿素b含量分别是对照的1.38和1.65倍。  【结论】  蚯蚓磨浆发酵产物的施入可有效提高土壤中细菌、放线菌数量,显著降低真菌数量和有害成分,改善苹果连作果园土壤环境,促进苹果幼树的生长,从而降低苹果连作障碍造成的危害。     

蚯蚓生物床工程处理对牛粪性质的影响

利用蚯蚓处理有机固体废弃物比传统技术手段具有更明显的优越性,得到越来越广泛的关注。试验通过蚯蚓生物床工程处理牛粪,采用室内分析的方法研究处理前后性质变化特征。结果表明：牛粪经蚯蚓生物床处理后的通水性和透气性得到明显改善,易干燥,无臭气,含水率和电导率明显下降,pH值趋于中性。蚯蚓生物床工程处理使全氮和有机质含量下降,提高了铵态氮、硝态氮、全钾、全磷、有机磷、无机磷和速效磷的含量,降低了钾的生物有效性,对腐殖酸含量无明显影响。对Cr、Cd和Cu的去除效果较好,对Pb、Mn、Zn和Fe的去除效果不佳。蚯蚓生物床工程处理过程丰富了蚯蚓粪中微生物区系,增加了过氧化氢酶活性,降低了脲酶活性,使牛粪中的碳水化合物发生逐步分解,生成腐殖物质,同时使水溶性有机硅化合物分解为无机硅氧化物。     

Mate choice of an endogeic earthworm revealed by microsatellite markers

Endogeic earthworms are difficult study subjects due to the cryptic medium in which they live; thus, only the behaviour of epigeic and anecic earthworms has been studied before. We used microsatellite markers as a tool to elucidate the mate choice processes of Hormogaster elisae, an endogeic earthworm. It was shown to normally mate with two partners, preferably of the same size that are found in close proximity thereby eliminating the need for long-distance dispersion, which could explain the previously observed high genetic differentiation between populations. The genetic analyses of the sperm within each of its four spermathecae showed a uniform distribution with no signs of differential storage of sperm from different partners.     

根区饲养蚯蚓对椪柑树体营养和根系生长的影响

以4年生桠柑为试验材料,研究根区饲养蚯蚓对椪柑树体营养和根系生长的影响。试验设4个处理,分别是穴施牛粪、穴施牛粪并接种蚯蚓、根系输液和对照。研究结果表明：穴施牛粪并接种蚯蚓很好地诱导了椪柑根系的生长,其诱导的根量比穴施牛粪和对照的分别高出100．62％和126．27％;穴施牛粪并接种蚯蚓处理的椪柑根系吸收和累积的氮、磷、钾元素分别比对照的高出21．13％、35．44％和76．31％;穴施牛粪并接种蚯蚓明显提高植株叶片的氮、磷、钾营养水平和叶绿素含量,在改善植株矿质营养状况方面优于穴施牛粪处理和根系输液处理。根系输液处理能在短时间内明显增加植株叶片营养含量。     

Behavioural endpoints in earthworm ecotoxicology

Background, Aims and Scope  Endpoints in earthworm ecotoxicology scheduled in guidelines are mortality and reproduction rates. However, not only the direct influence of pollutants on population parameters but also changes in behaviour such as substrate avoidance can have an important impact on soil ecosystems. In practice two different avoidance response tests are applied in earthworm ecotoxicology: (i) a six-chamber test system and (ii) a two-chamber test system. Both avoidance response-test systems were compared to establish their respective advantages and disadvantages in order to advance the standardisation of behavioural tests. The earthworm avoidance-response tests were applied in addition to the standard acute and chronic earthworm toxicity tests (ISO 11268) in order to compare the sensitivity of the test endpoints. Methods  Test substrates were contaminated with crude oil and 2,4,6-Trinitrotoluene (TNT), respectively. The test species wasEisenia fetida. The earthworms were exposed to the contaminated substrates and their mortality (14 d), reproduction rates (number of cocoons after 28 d, juvenile hatching after 56 d), and substrate preference (48 h) determined. Results and Discussion  Whereas 1000 mg/kg TPH (Total Petroleum Hydrocarbons) did not show any lethal effects, 100% mortality occurred in soil with comparable TNT concentration. The acute tests consistently produced the highest effect concentrations whereas reproduction and substrate avoidance were the more sensitive test parameters. Both behavioural test systems, when compared, showed similar substrate avoidance after an incubation time of 48 h. The six-chamber test system provides the potential to test six different substrates/concentrations at one time. It was observed, however, that earthworms did not migrate among all test chambers within a test unit in order to select the most appropriate substrate. Orientation was observed only between directly neighbouring test compartments, which complicates the interpretation of the test results. Conclusion  Substrate avoidance and reproduction variables were clearly more sensitive test endpoints than mortality. Therefore avoidance-response tests proved to be useful test methods in detecting effects of sublethal concentrations of pollutants on earthworms. The test duration of the avoidance tests is much shorter compared to the standard acute and chronic earthworm toxicity tests, which makes them a quick screening tool for identifying potential soil toxicity. Both avoidance-response test systems showed comparable results regarding the test sensitivity. Nonetheless, the incomplete substrate use in the six-chamber avoidance test due to the reduced migration possibilities (orientation only to neighbouring chambers) might reduce the distinctness of test results as it allows only reliable information on the most avoided and therefore most toxic substrate but not on 1 a clear dose-response pattern. Thus, to gain valid results, the number of replicates and the arrangement of the different substrates must be adopted. The two-chamber test system is less time-consuming due to easy handling and test results can be quantified more easily. Recommendations and Outlook  In consequence of the better validity of test results, lower expenses for test containers and less time for handling, the use of the two-chamber system is preferred over the six-chamber test system to assess the toxicity of polluted soil. Because of the ecosystem consequences of behavioural effects and the fact that avoidance response tests can reveal the toxic potential of pollutants in low concentrations, such tests should be included into ecotoxicological test protocols.     

Impacts of wetting-drying cycles on short-term carbon and nitrogen dynamics in Amynthas earthworm casts

Effects of earthworm casts on soil nutrient dynamics and their responses to changing moisture availability in subtropical ecosystems remain poorly understood. This study aimed to examine short-term carbon(C) and nitrogen(N) dynamics and their interactions with wetting-drying cycles in three different structural forms(i.e., granular, globular, and heap-like) of Amynthas earthworm casts. The rates of C and N mineralization in the earthworm casts were examined under two different wetting-drying cycles(i.e., 2-d and 4-d wetting intervals) using a rainfall simulation experiment. After three simulated rainfall events, subsamples of the earthworm casts were further incubated for 4 d for the determination of CO_2 and N_2O fluxes. The results of this study indicated that the impacts of wetting-drying cycles on the short-term C and N dynamics were highly variable among the three cast forms, but wetting-drying cycles significantly reduced the cumulative CO_2 and N_2O fluxes by 62%–83% and 57%–85%, respectively, when compared to the control without being subjected to any rainfall events. The C mineralization rates in different cast forms were affected by the amount of organic substrates and N content in casts, which were associated with the food preference and selection of earthworms. Meanwhile, the cumulative N_2O fluxes did not differ among the three cast forms. Repeated wetting and drying of casts not only enhanced aggregate stability by promoting bonds between the cast particles, but also inhibited microbial survival and growth during the prolonged drying period, which together hindered decomposition and denitrification. Our findings demonstrated that the interactions between the structural forms, aggregate dynamics, and C and N cycling in the earthworm casts were highly complex.