蚯蚓金属硫蛋白定量PCR检测方法及其分子诊断

根据GenBank中蚯蚓(Eisenia fetida)金属硫蛋白(MT)基因序列设计合成引物,采用RT-PCR扩增出目的基因片段,将目的基因片段插入pEASY-Blunt中制备质粒标准品.通过不同梯度稀释倍数的质粒标准品,建立了目的基因MT与内参基因β-actin的SYBR Green I荧光定量RT-PCR检测方法.通过土壤染毒培养实验,将蚯蚓分别暴露于50,500,1000mg/kg 重金属Cd染毒的自然土壤中培养14,28d,研究蚯蚓体内MT mRNA转录水平的表达变化.结果表明,重组质粒测序后显示目的片段序列同源性好,证明所设计的MT、β-actin引物能成功用于蚯蚓MT mRNA的检测.两基因构建的荧光定量标准曲线线性关系分别为0.994,0.999,且PCR扩增效率也皆接近于100%.染毒实验发现,Cd可诱导蚯蚓体内MT mRNA的表达,其表达水平与Cd污染暴露呈现出剂量-效应和时间-效应关系,表明蚯蚓MT基因可作为潜在分子标志物,诊断环境中Cd污染及其暴露水平.     

Earthworm Invasion as the Driving Force Behind Plant Invasion and Community Change in Northeastern North American Forests

Abstract:  Identification of factors that drive changes in plant community structure and contribute to decline and endangerment of native plant species is essential to the development of appropriate management strategies. Introduced species are assumed to be driving causes of shifts in native plant communities, but unequivocal evidence supporting this view is frequently lacking. We measured native vegetation, non-native earthworm biomass, and leaf-litter volume in 15 forests in the presence and absence of 3 non-native plant species ( Microstegium vimineum, Alliaria petiolata, Berberis thunbergii ) to assess the general impact of non-native plant and earthworm invasions on native plant communities in northeastern United States. Non-native plant cover was positively correlated with total native plant cover and non-native earthworm biomass. Earthworm biomass was negatively associated with cover of native woody and most herbaceous plants and with litter volume. Graminoid cover was positively associated with non-native earthworm biomass and non-native plant cover. These earthworm-associated responses were detected at all sites despite differences in earthworm species and abundance, composition of the native plant community, identity of invasive plant species, and geographic region. These patterns suggest earthworm invasion, rather than non-native plant invasion, is the driving force behind changes in forest plant communities in northeastern North America, including declines in native plant species, and earthworm invasions appear to facilitate plant invasions in these forests. Thus, a focus on management of invasive plant species may be insufficient to protect northeastern forest understory species.     

利用蚯蚓处理城市污水厂剩余污泥的初步试验研究

采用城市污水处理厂的剩余污泥（滤泥）和牛粪直接配置养殖蚯蚓的饲料,按照不同比例的配料分组进行了饲养蚯蚓的试验研究。在适宜的环境条件下,经过21 d培养,大平二号蚯蚓的最少增重为54％,最高增重达100％;平均产卵茧的数量为0．687个／条蚯蚓。研究结果表明,滤泥不需经过发酵即可用于养殖蚯蚓,牛粪与滤泥配置的比例为1∶1或0．3：0．7时,有利于蚯蚓的代谢和繁殖,并可确保蚯蚓养殖的安全。     

土壤甲苯、乙苯和二甲苯对蚯蚓及小麦的毒性效应

以赤子爱胜蚓(Eisenia fetida)和小麦(Triticum aestivum L.)为受试生物,研究了土壤中甲苯、乙苯和二甲苯等苯系物(TEX)的毒性效应.结果表明,3种污染物对赤子爱胜蚓和小麦毒性影响呈明显的剂量-效应关系,甲苯、乙苯和二甲苯对蚯蚓的24hLC50分别为583.6,346.8,192.4mg/kg;48h LC50分别为454.3,167.1,127.2mg/kg.甲苯、乙苯和二甲苯对小麦芽伸长的10%抑制率(IC10)分别为342.2,195.4,45.9mg/kg;对小麦根伸长的10%抑制率(IC10)分别为206.7,134.5,26.3mg/kg.小麦芽长、根长均可用于指示土壤被甲苯、乙苯及二甲苯污染的程度,但小麦种子根长对3种污染物胁迫的响应较芽长更为敏感,根长抑制率与芽长抑制率之间呈明显的相关关系.     

Single and Joint Effects of Acetochlor and Urea on Earthworm Esisenia Foelide Populations in Phaiozem

Much attention is paid to soil health and environmental safety. Earthworms are an important indicator of soil ecosystem health and safety. Ecological toxicity of acetochlor and excessive urea, in both their single and joint effects, on earthworm Esisenia foelide was thus studied using the soil-culture method. Acetochlor had an enhanced toxicity from low concentration to high concentration. The mortality of earthworms after a 6-day exposure was changed from 0 to 86.7%, and the weight change rate ranged from 7.86 to –30.43%, when the concentration of acetochlor was increased from 164 to 730 mg kg^–1. Urea expressed its positive and beneficial effects on earthworms when its concentration was lower than 500 mg kg^–1. Strongly toxic effects took place when the concentration of urea was higher than 1000 mg kg^–1. The mortality of earthworms exposed to urea reached 100% when its concentration was more than 1500 mg kg^–1. When the concentration of urea was lower than 500 mg kg^–1, there were antagonistic effects between the two agrochemicals on earthworms; when the concentration of urea was higher than 500 mg kg^–1, joint toxic effects of acetochlor and excessive urea on earthworms were synergic. In any case, excessive urea application is very harmful to the health of soil ecosystems.     

硫丹对蚯蚓存活、生长和肠胃线粒体超微结构的影响

通过人工土壤法对蚯蚓进行14d的急性暴露,研究了硫丹对蚯蚓存活、生长的影响;试验结束后,从硫丹剂量为0.5mg·kg-1和5 mg·kg-1的处理中挑选蚯蚓制作超薄切片,观察肠、胃部线粒体超微结构的变化.结果表明,硫丹对蚯蚓有较强的毒性作用,14d的LC50为6.52mg·kg-1.试验期间硫丹对蚯蚓的生长有显著的抑制作用,生长抑制率随硫丹剂量和暴露时间增加而增大.蚯蚓肠胃部的线粒体对硫丹有较强的敏感性,在非致死剂量下超微结构的损伤程度随硫丹剂量增加而加重.     

蚯蚓在污水土地处理中的应用分析

蚯蚓是一种重要的资源,一方面为人类提供蛋白质、氨基酸等营养;另一方面作为分解者,可以用来治理或降低环境污染,保护生态环境。污水土地处理具有低建设成本、低运行成本和管理方便等特点而引起了广泛关注,但土地处理也存在占地面积比较大,氮、磷去除率不高,土壤堵塞和运行不稳定等问题。针对这些问题,文章分析了蚯蚓在污染物去除、提高植物生物量和提高土壤渗透率等方面的作用,并分析了蚯蚓在解决污水土地处理中土壤堵塞、植物退化、水力负荷比较低和占地面积比较大等问题的可行性,提出将蚯蚓引入土地处理系统能提高土地处理的效率和运行的稳定性,加快污水土地处理技术的应用。     

超声波对污泥蚯蚓堆肥有机质和重金属的影响

联合超声波破解技术、好氧堆肥技术及蚯蚓处理技术,对污水处理厂剩余污泥进行处理,探讨超声波预处理对污泥蚯蚓堆肥有机质降解和重金属的影响.结果表明：超声波预处理技术对污泥腐殖化程度表现出协同效应,最优TOC反应动力学常数为0.0084d-1,是对照组的2.1倍;胡敏酸和HA/FA平均增长率分别为97.4%和191.5%,是对照组的1.26倍和1.37倍;可轻度减少污泥堆体重金属含量,同时对蚯蚓处理阶段重金属富集存在缓减效应,但对各阶段重金属形态影响不显著;蚯蚓处理可明显降低重金属生物有效性,至处理结束Zn、Cu、Pb、Ni、Cd、Cr、Hg、As非稳定态平均占比较好氧堆肥结束时下降28.3%、29.5%、29.1%、35.2%、3.3%、25.3%、36.4%、21.9%.     

The rhythm of stereotypic behaviors in captive giant panda (Ailuropoda melanoleuca)北大核心CSCD

Stereotypy can be defined as a repetitive behavioral pattern with no apparent function or target in captivity. It is considered an abnormal behavior in contrast to the behavior in wild habitats and is often used as an important index to evaluate the captive environment, which has a significant scientific value. Many previous studies have focused on the impact of environment on stereotypical behavior; however, the rhythm of stereotypy remains unclear. To determine the mechanism of stereotypy, eight giant pandas (Ailuropoda melanoleuca) were investigated 24 h a day from Jan 2018 to Dec 2019, with focal sampling and continuous recording. No stereotypy in the juveniles was noted, while the adult giant pandas displayed different stereotypies. Mean stereotypy durations were 0.72% of the total activity in circadian rhythm, with a peak of mean durations in all seasons occurring at 7:00 am–9:00 am, while, in spring, there was a peak of mean duration at 1:00 am–3:00 am. Although there was no significant difference, mean stereotypy durations and frequencies in the night were longer than those in the daytime. Mean stereotypy frequencies in spring reached the highest level, whereas mean stereotypy durations in winter were maximal. The adult giant pandas in the daytime showed maximal mean stereotypy durations in winter and minimal mean stereotypy durations in busy seasons (summer and autumn), whereas mean stereotypy durations at night were maximal in busy seasons. Despite no significant differences among these seasons, our results suggest that the stereotypy of giant pandas may have diurnal rhythms and seasonal differences. In light of the similar captive environment between adult and juvenile giant pandas, the appearance of stereotypy may be correlated with age, which may be the effects of the environment or a strategy of adaptation. © 2022 Science Press. All rights reserved.     

低繁殖量蚯蚓养殖法处理剩余污泥的可行性研究

针对城市剩余污泥饲养蚯蚓需要发酵预处理,以及繁殖的含重金属的蚯蚓利用难的问题,进行了不同种类污泥直接饲养蚯蚓实验,并考察了投料方式、投料厚度对蚯蚓的生长影响.结果显示,新鲜的城市剩余污泥直接饲养蚯蚓可行,与投料方式无关.发臭污泥块状投放适宜蚯蚓生长,密实投放不适宜蚯蚓生存.采用定时引出成蚓,改变泥中蚓卵的生存环境,控制卵发育的养殖方法,对污泥中重金属转移规律进行了研究.结果表明,饲养过程中蚯蚓体内的重金属质量比随饲养时间上升.饲养120 d左右,蚯蚓体内质量比达到极限,且处理后的污泥中重金属质量比明显下降.该养殖法处理剩余污泥是方法可行,且具有蚯蚓产量低,利于蚯蚓处理和处置的优点.