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。     

春季树麻雀体内几种消化酶活性研究

应用酶学分析法测定了树麻雀Passer montanus春季的腺胃、肌胃、小肠、大肠、肝脏和胰脏组织中淀粉酶、蛋白酶和纤维素酶的活力.结果表明,不同组织中的消化酶活力差异显著,淀粉酶和蛋白酶以胰脏中活力最高,腺胃次之,纤维素酶均较低;同一组织中不同消化酶的活力差异显著,淀粉酶活性最高,蛋白酶次之,纤维素酶活力最低.这些差异提示消化酶活力大小与器官分化有关,并受食物组成的影响,因此产生了不同的酶活力分布.这是树麻雀长期适应东北地区寒冷环境的生存策略之一.     

Molecular cloning and characterization of cDNA encoding fibrinolytic enzyme-3 from earthworm Eisenia foetida

Earthworm fibrinolytic enzyme (EFE), a multi-com-ponent protease purified from some earthworm breeds,belongs to serine protease family with fibrinolytic activity[1]. It has been used in prevention and treatment of cardiacand cerebrovascular diseases in Ch…     

Crystal structure of earthworm fibrinolytic enzyme component a: revealing the structural determinants of its dual fibrinolytic activity

Earthworm fibrinolytic enzyme component A (EFEa) from Eisenia fetida is a strong fibrinolytic enzyme that not only directly degrades fibrin, but also activates plasminogen. Proteolytic assays further revealed that it cleaved behind various P1 residue types. The crystal structure of EFEa was determined using the MIR method and refined to 2.3A resolution. The enzyme, showing the overall polypeptide fold of chymotrypsin-like serine proteases, possesses essential S1 specificity determinants characteristic of elastase. However, the beta strand at the west rim of the S1 specificity pocket is significantly elongated by a unique four-residue insertion (Ser-Ser-Gly-Leu) after Val217, which not only provides additional substrate hydrogen binding sites for distal P residues, but also causes extension of the S1 pocket at the south rim. The S2 subsite of the enzyme was partially occluded by the bulky side-chain of residue Tyr99. Structure-based inhibitor modeling demonstrated that EFEa's S1 specificity pocket was preferable for elastase-specific small hydrophobic P1 residues, while its accommodation of long and/or bulky P1 residues was also feasible if enhanced binding of the substrate and induced fit of the S1 pocket were achieved. EFEa is thereby endowed with relatively broad substrate specificity, including the dual fibrinolysis. The presence of Tyr99 at the S2 subsite indicates a preference for P2-Gly, while an induced fit of Tyr99 was also suggested for accommodation of bigger P2 residues. This structure is the first reported for an earthworm fibrinolytic enzyme component and serine protease originating from annelid worms.     

蚯蚓纤溶酶分子生物学进展

蚯蚓纤溶酶是近年发现的一种新型的溶解血栓物质,属丝氨酸蛋白酶,不同种属的蚯蚓中均可分离到,具纤溶活性和溶栓活性。有较好的热稳定性,多为单体酶,多数兼有纤溶活性和纤溶酶原激活活性。不同种属的蚯蚓分离的纤溶酶性质上有一定差别。已获得多种纤溶酶的N端序列及部分核酸序列,相互之间及与某些蛋白酶之间有一定的同源性。纤溶酶通过降解目的蛋白的特定位点而起作用 。     

Toxicokinetics and Biotransformation of As(III) and As(V) in Eisenia fetida

Earthworms, Eisenia fetida, were exposed to soils spiked with As(III) and As(V), to understand the response of earthworms to As in terms of both toxicity and accumulation using toxicokinetics, and to explain As metabolism and bioavailability. As(III) showed higher toxicity than As(V), in all toxicity endpoints of burrowing time, survival, growth, and cocoon production. Bioconcentration occurred at both As(III) and As(V) treatments during 28-d exposure. Worms did not show the elimination of As for consecutive 28 d of exposure to clean soils. Biotransformation of As was characterized using HPLC-ICP-MS and XANES, showing the reduction of As in worms regardless of the As species to which worms were exposed. Metabolism of As in worms that formed As-thiol complex is thought to limit the excretion of As, and thus induce bioconcentration in worms. Uptake rates by one-compartment model indicated that pore water was the bioaccessible pool of As, and directly controlled the uptake of As by worms. The study suggests that higher uptake rate and bioaccumulation of As(III) than of As(V) are among the factors that make As(III) more toxic than As(V).     

Bioremediation of Agricultural Wastes through Vermicomposting

The post-harvest residues of some local crops, e.g., wheat (Triticum aestivum), millets (Penniseum typhoides and Sorghum vulgare), and a pulse (Vigna radiata) were subjected to recycle through vermicomposting. The crop residues were amended with animal dung, and three types of vermibeds were prepared: (i) millet straw (S. vulgare + P. typhoides in equal quantity) + sheep manure (1: 2 ratio) (MS); (ii) pulse bran (V. radiata) + wheat straw (T. aestivum) + cow dung (1: 1: 2 ratio) (PWC); and (iii) mixed crop residues (mixing of all types crop residues, used in this study) + cow dung in 1:1 ratio (MCR + CD). The fourth treatment was cattle shed manure (CSM). Vermicomposting resulted in a significant increase in total N (97.3% to 155%), available P (67.5% to 123.5%), exchangeable K (38.3% to 112.9%), and exchangeable Ca (23.3% to 53.2%), and decrease in organic C content (20.4% to 29.0%) in the different vermibeds. The earthworm showed the higher biomass gain, growth rate (mg wt. worm^?1 day^?1) and cocoon numbers in the CSM vermibed. The quality of crop residues was directly related to the rate of organic matter mineralization during the vermicomposting. This study suggests that agriculture wastes could be converted into some value-added products, e.g., vermicompost and worm biomass through vermicomposting technology. The higher concentrations of plant nutrients in end products indicate a potential for using agriculture wastes in sustainable crop production.     

蚯蚓纤溶酶基因的克隆与序列分析

目的:获取蚯蚓纤溶酶(EFE)的基因,并对其进行序列分析。方法:从赤子爱胜蚓中提取总RNA,然后利用RT-PCR技术扩增EFE的基因,并将其插入pUCm-T载体,经PCR和酶切鉴定后进行序列测定及分析。结果:DNA序列分析表明,所克隆的EFE基因全长为738bp,其中编码区段为735bp,共编码245个氨基酸残基,成熟肽为238个氨基酸残基。与GenBank中已报道的粉正蚓的EFE序列F-Ⅲ-2的同源性最高,两者在核苷酸序列上有2处不同,即第137位(T→C)和第632位(G→T),密码子也因而分别由GTC、GGT变为GCC、GTT,导致第46位、211位的氨基酸残基分别由缬氨酸、甘氨酸变为丙氨酸和缬氨酸。结论:从赤子爱胜蚓中成功克隆了1条EFE基因F245。序列测定及同源性分析表明,蚯蚓纤溶酶F245与已报道的多条EFE基因序列具有高度同源性,且具备完整的编码区。该序列的克隆为用基因工程的方法生产单一成分的高酶活性EFE奠定了基础。