不同生活型蚯蚓对污泥处理的效果研究

研究报道了三种不同种蚯蚓接种后污泥的处理效果。28 天的培养实验表明: 接种蚯蚓, 特别是体型较大的威廉环毛蚓和杜拉蚓能提高污泥的pH 值, 减少污泥中有机物的含量。接种蚯蚓减少了污泥中全氮、全磷、全钾的含量, 深层种杜拉蚓显著降低了污泥中全氮和全磷含量, 较对照处理分别减少了14.3%和10.3%。接种威廉环毛蚓的处理较对照处理能显著增加水解性氮和速效钾的含量。接种杜拉蚓的处理吸收重金属的效果最佳, Cr 和Cu 含量均显著低于对照处理, 威廉环毛蚓次之, 但优于赤子爱胜蚓。结果显示, 在大部分项目中, 杜拉蚓和威廉环毛蚓的作用效果是要优于赤子爱胜蚓, 不同生活习性的蚯蚓对污泥净化效果会有着一定的不同。     

两种代表性蚯蚓对设施菜地土壤微生物群落结构及理化性质的影响

不同生态型蚯蚓的取食偏好和生境有所差异,因此蚯蚓的生态型差异可能关乎其对土壤性质的不同影响;有关不同生态型蚯蚓对土壤性质尤其是微生物学性质影响的研究有助于了解蚯蚓生态功能的作用机制。在野外调控试验的第4年采集土壤,研究了牛粪混施和表施处理下内层种威廉腔环蚓(Metaphire guillelmi)和表层种赤子爱胜蚓(Eisenia foetida)对设施菜地土壤微生物群落结构和主要理化性质的影响。结果表明,土壤微生物群落结构同时受到蚯蚓种类和牛粪施用方式的影响。牛粪表施时,两种蚯蚓均显著降低了菌根真菌、真菌生物量和原生动物生物量(P0.05);牛粪混施时,不同蚯蚓的影响有所差异,威廉腔环蚓明显增加了菌根真菌、真菌生物量和放线菌生物量,而赤子爱胜蚓的作用不明显。此外,两种蚯蚓均提高了土壤孔隙度、团聚体稳定性和土壤p H、矿质氮以及微生物生物量碳氮水平,但提高幅度取决于蚯蚓种类和牛粪施用方式。冗余分析表明蚯蚓影响下土壤微生物群落结构的变化与团聚体稳定性、p H、速效磷、矿质氮呈正相关,而与土壤容重呈负相关。     

4种蚯蚓肠道微生物对砷毒性的响应差异研究

蚯蚓肠道是微生物多样性的一个潜在存储库。砷对蚯蚓肠道微生物群落的影响已被证实,但砷在不同蚯蚓肠道菌群中生物转化的差异仍不清楚。为了进一步阐述土壤中广泛存在的低浓度砷(浓度为5,15,25 mg/kg)对不同种类蚯蚓肠道微生物影响的差异,将4种典型蚯蚓暴露于砷污染土壤后,测定其肠道微生物组成变化,并分析砷对不同蚯蚓肠道内砷富集、形态和砷生物转化基因的影响。结果显示,所有蚯蚓组织内均存在明显的砷富集,其富集系数由高到低依次为:安德爱胜蚓(1.93)>加州腔蚓(0.80)>通俗腔蚓(0.78)>湖北远盲蚓(0.52),蚯蚓组织和肠道内砷形态主要以无机砷为主,其中As(III)含量比例> 80%,部分蚯蚓组织内还发现少量有机砷。4种蚯蚓肠道微生物群落在门水平上主要以变形菌、厚壁菌和放线菌为主,并与周围土壤细菌群落组成存在显著差异。同时,在土壤和肠道内共检测到17个砷转化基因,其中蚯蚓肠道内As(V)还原和砷转运相关基因相对丰度较高,而砷(去)甲基化基因丰度较低。此外,低浓度砷污染对蚯蚓生长无显著影响,却能引起蚯蚓肠道微生物群落的紊乱。蚯蚓种类和砷污染是引起蚯蚓肠道微生物...     

白颈环毛蚓（Pheritima catifornica）与壮伟环毛蚓（Amynthas f…

对白颈环毛蚓与壮伟环毛蚓的体壁及内脏器官ＰＴＣ－氨基酸的含量进行了初步分析,结果表明;两种蚯蚓具有１５种氨基酸,８种必需氨基酸含量均较高。而壮伟环毛蚓的必需氨基酸总量均超过白颈环毛蚓,也大大超过国产或进口于粉氨基酸总含量;加上壮伟环毛蚓个体大,分布广,又耐污染,抗病能力强,所以这量种值得开发的动物蛋白的新资源。     

西双版纳热带次生林和橡胶林蚯蚓数量分布及其与环境因子的关系

2006年1月,通过样地调查法比较了西双版纳次生林和橡胶林2种不同土地利用方式的蚯蚓数量分布状况及其与环境因子的关系.结果表明:自然林转变为橡胶林后,土壤养分及植物有机碳输入均明显减少,同时蚯蚓群落发生较大的变化:在橡胶林,热带外来种蚯蚓无围海蚓的密度(52.27 ind·m-2)显著高于林地内本地种蚯蚓远环蚓的密度(0.53 ind·m-2),但在次生林,2种蚯蚓的密度和生物量无显著差异;橡胶林本地种蚯蚓远环蚓的密度(0.53 ind·m-2)和生物量(0.32 g·m-2)与次生林(6.93 ind·m-2、7.76 g·m-1)相比呈下降趋势.无围海蚓的幼蚓在其种群中占有较大比例,类似增长型种群年龄结构,而远环蚓则主要以成蚓为主.相关分析表明,橡胶林的细根生物量与无围海蚓的密度呈显著正相关.由预示蚯蚓群落结构的这种变化可能与橡胶林土地利用引起的植被变化、土壤养分下降等因素密切相关,同时外来种蚯蚓与本地种蚯蚓本身的生态适应对策也不可忽视.     

峨眉山大蚯蚓肠道可培养细菌多样性研究

为探究峨眉山大蚯蚓秉前环毛蚓(Pheretima carnosa)肠道可培养细菌多样性,测定采样点土壤基本理化性质,从蚯蚓肠道中分离可培养细菌并进行16S r DNA系统发育分析,探究分离菌株的相似性、多样性、优势度及均匀度,绘制稀释曲线。结果显示:峨眉山大蚯蚓生活在有机质丰富的土壤环境中,其肠道中分离出的56株细菌分布在3门14属,其中芽孢杆菌属(Bacillus)为明显优势菌群;此外,Shannon-Wiener多样性指数(1.73)、Berger-Parker优势度指数(0.536)、Margalef指数(3.23)、Evenness均匀度指数(0.66)均表明峨眉山大蚯蚓肠道可培养细菌多样性丰富,而且稀释曲线表明尚有大量未分离细菌。上述研究为峨眉山大蚯蚓肠道微生物资源的开发利用奠定了基础。     

四川陆栖寡毛类两新种记述：寡毛纲：链胃蚓科,棘蚓科

记述了采集于四川省酉阳县的链胃科、杜拉属Drawida蚯蚓一新种,双斑杜拉蚓Drawida bimaculata sp.nov.和在四川省汉源县采到的棘蚓科,毛蚓属Plutellus蚯蚓一新种,汉源毛蚓Plutellus hanyuangensis sp.nov.。毛蚓属是我国首次记录。     

毛苔草湿地土壤酶活性及活性有机碳组分对水分梯度的响应

通过设置的W1（15cm）、W2（-5cm）、W3（-5～5cm）、W4（淹没）4种水分梯度的毛苔草（Carex lasiocarpa）盆栽培养实验,研究了湿地土壤酶活性、微生物量碳（MBC）、可溶性有机碳（DOC）及毛苔草地上生物量对水分梯度的响应及土壤酶活性与MBC、DOC、地上生物量的关系。土壤酸性磷酸酶、蔗糖酶和脲酶活性随着土壤水分增加而降低,但过氧化氢酶活性随着土壤水分增加而增加。与持续淹水相比,于湿交替（W3）增加了土壤酸性磷酸酶、蔗糖酶、脲酶和过氧化氢酶活性。土壤微生物量碳（MBC）含量表现为W3〉W1〉W2〉W4,蔗糖酶、脲酶、过氧化氢酶活性与MBC呈显著正相关（P〈0．05）。土壤可溶性有机碳（DOC）含量表现为W4〉W1〉W3〉W2,脲酶和酸性磷酸酶活性与DOC呈极显著负相关（P〈0．01）。毛苔草地上生物量与土壤酶活性呈正相关,其中,蔗糖酶、过氧化氢酶、脲酶活性与毛苔草生长状况密切相关。     

用COI基因确定直隶环毛蚓(Pheretima tschiliensis tschiliensis)的属的地位

目的:由于采用新的分类体系,国内外学者通过形态学特征对直隶环毛蚓(Pheretima tschiliensis tschiliensis,Michealson,1928)的分类地位一直存在争议,该文的目的在于将此种给予明确的划分。方法:用采自不同地区的直隶环毛蚓COI基因(534bp)及结合其相应的氨基酸(178个),以陆正蚓(Lumbricus tereistial)为外群,用最大简约法和贝叶斯法构建进化树的方法,结果:试验结果表明,构建的系统树将直隶环毛蚓与腔蚓属聚为一族,并得到很好的支持(BP>76,pp≥0.94),种间校正基因距离介于0.006~0.263,种内校正基因距离介于0.000~0.162,结论:因具有交配腔,故分子生物学和形态学特征均表明,直隶环毛蚓应属于腔蚓属而不是远盲蚓属。其学名应为Metaphire tschiliensis tschiliensis(Michealson,1928)。     

土壤温度和湿度对外来种蚯蚓Pontoscolex corethrurus 产茧和幼蚓孵化的影响

通过采用主处理为土壤温度,副处理为土壤湿度的裂区区组实验设计,在室内微生态条件下控制土壤温度和湿度培养蚯蚓,探讨了3种土壤温、湿度对热带外来种蚯蚓Pontoscolex corethrurus成蚓产茧和蚓茧孵化的影响。研究结果表明,P. corethrurus成蚓在20—30℃土壤温度范围、25%—35%土壤湿度环境中均可持续产茧和孵化幼蚓,培养时间、土壤温湿度对蚯蚓的产茧和孵化呈现显著影响。20%土壤湿度下成蚓休眠或死亡。35%土壤湿度下呈现最大蚓茧产量和幼蚓孵化量。在较好（35%）土壤湿度下产茧数表现为高温好于低温,反之亦然;在适宜（25%—35%）土壤湿度下幼蚓孵化率随温度升高而增加,但呈现高、低温的限制作用,即25℃土壤温度和35%土壤湿度时出现最高幼蚓孵化率。在适宜的土壤温湿度范围内,湿度较温度对蚯蚓繁殖具有更显著的控制作用,温度的影响在一定程度上可通过土壤湿度加以调节。     

Bt毒蛋白Cry1Ac在人造土壤中对赤子爱胜蚓毒理及生化影响

Bt毒素能通过转基因作物的花粉、根和残株进入土壤.为评估转基因作物对土壤动物的影响,本文模拟转基因棉的Bt毒素进入土壤的发生程度,用含不同浓度Bt毒蛋白Cry1Ac的人造土壤处理蚯蚓,测定蚯蚓存活率、重量变化及体内总蛋白含量和过氧化氢酶(CAT)、乙酰胆碱酯酶(AchE)、谷胱甘肽-S-转移酶(GST)和纤维素酶活性.结果表明,Bt毒蛋白对蚯蚓的生物量和生理水平影响均不明显,不存在急毒性和亚致死毒性影响,对蚯蚓比较安全.     

蚯蚓粪对黄瓜苗期土传病害的抑制作用

试验表明,蚯蚓对农业有机废弃物进行生物降解的产物－蚯蚓粪,在一定程度上能够控制蔬菜类黄瓜苗期土传病害的发生,并表现出明显的促生长效应,蚯蚓粪控制病害的程度与蚯蚓粪的量有一定的关系,当蚯蚓粪与土体积比为20％时,控制病的程度最大,防效达96．1％,这种作用主要与蚯 蚓粪中的微生物性质有关,蚯蚓粪能大大提高土壤中的微生物量和微生物活性,从而大大增强了病土中与病原菌进行能源竞争的微生物的竞争能力,同时从新鲜蚯蚓粪中成功分离到拮抗活性强,抗菌谱广的拮抗微生物,初步研究结果说明,一般性抑制和特殊性抑制两种机制在蚯蚓粪对病害的控制中起作用。     

Comparative toxicity of ten organic chemicals to four earthworm species

Ten organic chemicals were tested for toxicity to four earthworm species: Allolobophora tuberculata, Eisenia fetida, Eudrilus eugeniae and Perionyx excavatus, using the European Economic Community's (EEC) earthworm artificial soil and contact testing procedure. The phenols were the most toxic chemicals tested, followed by the amine, substituted benzenes, halogenated aliphatic hydrocarbon, polycyclic aromatic hydrocarbon and phthalate as the least toxic chemical tested. Correlations among species within each type of test for a given chemical were extremely high, suggesting that the selection of earthworm test species does not markedly affect the assessment of a chemical's toxicity. The correlation between the two tests was low for all test species. The contact test LC50 for a given chemical cannot be directly correlated to an artificial soil test LC50 for the same earthworm species.     

不同有机物料的蚯蚓堆肥及可溶性有机物的三维荧光光谱特征

研究了牛粪、茶渣、药渣和菌渣等有机物料的不同复配比例的蚯蚓生长状况,筛选出适合蚯蚓生长的最佳物料配比,并进一步分析了这些适合蚯蚓生长的物料配比处理的养分含量和三维荧光光谱特征的动态变化.结果表明： 牛粪与药渣、牛粪与茶渣的不同配比物料中的蚯蚓生长情况均较好,而有菌渣复配的物料的蚯蚓生长情况较差;随着堆肥时间的增加,堆肥的pH逐步趋向中性和弱酸性,堆肥的电导率、全氮、全磷、速效氮和速效磷含量逐渐增加,全钾和速效钾呈现出先增加后降低的趋势,而堆肥的有机质含量逐渐减小;三维荧光光谱特征和荧光区域指数结果表明,随堆肥时间增加,可溶性有机物(DOM)中的类蛋白峰逐渐减弱,而类腐殖质峰逐渐增强.蚯蚓堆肥改变了有机废弃物的DOM组成,生成了大量类胡敏酸和类富里酸物质, 性质逐渐趋于稳定.因此,有机废弃物蚯蚓堆肥中DOM物质含量变化可作为蚯蚓堆肥成熟度的评估指标.     

Crystal structure of earthworm fibrinolytic enzyme component B: a novel, glycosylated two-chained trypsin

The earthworm fibrinolytic enzyme (EFE), belonging to a group of serine proteases with strong fibrinolytic activity, has been used in a mixture as an oral drug for prevention and treatment of thrombosis in East Asia. The EFE component b (EFE-b) is one of seven EFE components from Eisenia fetida, and among them it has nearly the highest fibrinolytic activity. Here, we report its crystal structure at a resolution of 2.06A. The structural analysis shows that EFE-b should be classified as a trypsin from earthworm. However, it is distinct from other trypsins. It is a two-chained protease with an N-terminal, pyroglutamated light chain and an N-glycosylated heavy chain. Furthermore, the heavy chain contains a novel structural motif, an eight-membered ring resulting from a disulfide bridge between two neighboring cysteine residues, and a cis peptide bond exists between these two cysteine residues. The crystal structure of EFE-b provides the structural basis for its high level of stability and reveals its complicated post-translational modifications in earthworm. This structure is the first reported for a glycosylated two-chained trypsin, which may provide useful clues to explain the origin and evolution of the chymotrypsin family.     

Earthworms use odor cues to locate and feed on microorganisms in soil

Earthworms are key components of temperate soil ecosystems but key aspects of their ecology remain unexamined. Here we elucidate the role of olfactory cues in earthworm attraction to food sources and document specific chemical cues that attract Eisenia fetida to the soil fungi Geotrichum candidum. Fungi and other microorganisms are major sources of volatile emissions in soil ecosystems as well as primary food sources for earthworms, suggesting the likelihood that earthworms might profitably use olfactory cues to guide foraging behavior. Moreover, previous studies have documented earthworm movement toward microbial food sources. But, the specific olfactory cues responsible for earthworm attraction have not previously been identified. Using olfactometer assays combined with chemical analyses (GC-MS), we documented the attraction of E. fetida individuals to filtrate derived from G. candidum colonies and to two individual compounds tested in isolation: ethyl pentanoate and ethyl hexanoate. Attraction at a distance was observed when barriers prevented the worms from reaching the target stimuli, confirming the role of volatile cues. These findings enhance our understanding of the mechanisms underlying key trophic interactions in soil ecosystems and have potential implications for the extraction and collection of earthworms in vermiculture and other applied activities.     

Isolation and some characterizations of a glycosylated fibrinolytic enzyme of earthworm,Eisenia fetida

Resin coupled with m-aminophenylbornic acid was used to isolate a glycosylated component from homogenate of earthworm (Eisenia fetida). The fraction showed a single band on SDS-PAGE with a molecular weight of 34, 193 Da determined by mass spectroscopy. The N-terminal region is AQVCCPDI, different from those of earthworm fibrinolytic enzymes reported previously (Nakajima et al. 1993). This glycosylated component showed an activity on digesting both Chromozym-TH and fibrin, suggesting that it is a novel fibrinolytic enzyme.     

8-hydroxydeoxyguanosine generated in the earthworm Eisenia fetida grown in metal-containing soil

Heavy metal pollution of soil causes biological problems, such as mutagenicity to living organisms, including human beings. However, few methods have been developed to assess metal mutagenicity in soil. To avoid metal mutagenicity, an adequate bio-monitoring method is required. In the present study, to determine if the analysis of oxidative DNA damage generated in the earthworm is a useful bio-monitoring method for soil mutagenicity, the accumulation of 8-hydroxydeoxyguanosine (8-OH-dG), a major form of oxidative DNA damage, in Eisenia fetida (Savigny, 1826) treated with cadmium chloride (CdCl2) or nickel chloride (NiCl2) was analyzed. E. fetida was treated with Cd (10 or 200 microg/g soil) or Ni (10 or 200 microg/g soil) for 1, 2, and 3 weeks or 3 months. After metal exposure, the metal concentration in E. fetida was analyzed by atomic absorption spectrometry and the 8-OH-dG accumulated in E. fetida was analyzed by HPLC analyses and immunohistochemistry. Atomic absorption spectrometry revealed that Cd, but not Ni, accumulated within E. fetida. The 8-OH-dG levels in the DNA of E. fetida treated with Cd for 3 months were significantly higher than those in control E. fetida. Moreover, immunohistochemical analyses revealed that positive signals for 8-OH-dG accumulation in seminal vesicles were detected only in E. fetida treated with 10 microg of Cd for 3 months. Although some points remain unresolved, a bio-monitoring system analyzing the DNA damage generated in the earthworm might be useful for the assessment of the mutagenicity of soil contaminated with various heavy metals, such as Cd.     

Decomposition of kitchen waste amended with cow manure using an epigeic species (Eisenia fetida) and an anecic species (Lampito mauritii)

An epigeic (surface dweller) earthworm species Eisenia fetida and an anecic (deep burrower) earthworm species Lampito mauritii have been tested for decomposition of kitchen waste plus cow dung. Chemical analyses of worm-worked substrates by both species showed g/kg increases in nitrogen, phosphorus and potassium and decreases in C/N and C/P ratios after 150 days of vermicomposting. However, organic carbon matter showed reduction in their amounts for 3-4 months and afterwards slightly increased up to 150 days. E. fetida produced 0.27%, 156%, 41% and 38% increases in organic carbon, nitrogen, phosphorus, and potassium as well as 61% and 29% decreases in C/N and C/P ratios as compared to control after 150 days of earthworm inoculation. In contrast, L. mauritii produced 14%, 102%, 33% and 42% increases in organic carbon, nitrogen, phosphorus and potassium as well as 43% and 14% decreases in C/N and C/P ratios as compared to control after 150 days of earthworm activity. There was moderate mineralization and faster decomposition by E. fetida in comparison to moderate mineralization and moderate decomposition by L. mauritii. The average numbers of cocoons and adults produced were greater by E. fetida than by L. mauritii after 150 days. These results indicate E. fetida may be a better adapted species for decomposition of kitchen waste plus cow dung under tropical conditions.     

Eisenia fetida中蚯蚓纤溶酶P-Ⅲ组分序列多态性

为了探讨蚯蚓纤溶酶P—Ⅲ组分序列多态性及其与蚓种关系。从Eisenia fetida中提取总RNA,利用P-Ⅲ组分序列两端的保守性设计引物,通过RT—PCR获得P—Ⅲ组分基因,并构建该组分质粒（pUC18）文库,经PCR筛选后进行序列测定及分析。在原已获得的Efp-Ⅲ-1和Efp-Ⅲ-3的基础上,利用中间引物从文库中筛选到Efp-Ⅲ-2基因。序列分析表明,该基因编码序列为720bp,编码239个氨基酸,与GenBank报道的点Efp-Ⅲ-2五条序列整体相似性高达99．02％。通过序列比对,发现Efb-Ⅲ-1、2和3具有完全相同的结构域,差异位点零星分布在序列中间;Lumbricus rubellus的六条序列同样具有Eisenia fetida这个特点,并无明显差异。序列比对证明它们是蚯蚓体内P-Ⅲ组分基因多态性的表现,而且在Lumbricidae科中与蚓种无关。