两种新型杀虫剂在不同暴露系统对蚯蚓的急性毒性

１　引　言蚯蚓是土壤生态系统的一个组成部分，它作为非靶陆生土壤生物，能改善土壤的通气性，增进土壤肥力，同时，蚯蚓是鸟类等许多动物的食物源，在食物链中，它是陆生生物与土壤生物之间传递的桥梁［１］。农药的大面积施用，必将对蚯蚓的生存、生长、繁殖产生种种不利影响，导致土壤生态系统的变化，因此，研究农药对蚯蚓的毒性，是评价农药对生态环境安全性的一个重要指标。采用农药的急性毒性作为指标，对农药毒性进行评价，国内外均有报道［２～１１］。新型杀虫剂吡虫啉和抑食肼已经或正在投入生产，并将陆续进入市场，虽已根据农…     

土壤重金属污染对蚯蚓的急性毒性效应研究

测定了草甸棕壤条件下 ,Cu、Zn、Pb、Cd单一 /复合污染对蚯蚓的急性致死及亚致死效应 .结果表明 ,Cu、Pb浓度与蚯蚓死亡率显著相关 (α=0 .0 5 ,RCu=0 .86 ,RPb=0 .87) ,Cu浓度与生长抑制率显著相关 (α=0 .0 5 ,RCu=0 .84) ,其他供试重金属浓度与蚯蚓死亡率和生长抑制率相关性不显著 .蚯蚓个体对重金属毒性的耐受程度差别较大 .其毒性阈值 (引起个体蚯蚓死亡浓度 )分别为 :Cu 30 0mg·kg-1,Zn 130 0mg·kg-1,Pb 170 0mg·kg-1,Cd 30 0mg·kg-1.LC50 分别为 :Cu 40 0～ 45 0mg·kg-1,Zn15 0 0～ 190 0mg·kg-1,Pb2 35 0～ 2 40 0mg·kg-1,Cd 90 0mg·kg-1.在Cu、Zn、Pb、Cd单一污染引起 >10 %蚯蚓死亡的浓度下 ,复合污染导致 10 0 %蚯蚓死亡 ,表明复合污染极强的协同效应 .     

高铜、高锌猪粪对蚯蚓的急性毒性效应研究

测定了高Cu、高Zn猪粪条件下Cu、Zn单一与复合污染对蚯蚓的急性致死及亚致死效应.结果表明,Cu、Zn浓度与蚯蚓死亡率显著正相关(a=0.05,r_Cu=0.99,r_Zn=0.99),与体重增长率显著负相关(a=0.05,r_Cu=-0.99,r_Zn=-0.96).蚯蚓个体对Cu、Zn的耐受程度不同,其毒性阈值(引起蚯蚓个体死亡浓度)分别为:Cu250mg·kg^-1、Zn400mg·kg^-1.LD₅₀分别为:Cu646.68mg·kg^-1、Zn947.38mg·kg^-1.复合污染情况下,Cu浓度为250、500mg·kg^-1时,Cu、Zn复合污染表现为协同效应;Cu浓度为750mg·kg^-1时,Cu、Zn复合污染表现为拮抗效应,可见,猪粪中Cu、Zn复合污染的毒性效应与各组浓度组合密切相关.     

PFOS对蚯蚓急性毒性和回避行为的影响

全氟辛烷磺酰基化合物(PFOS)作为一种新型持久性有机污染物,已经成为环境科学和毒理学的研究热点,其对生态环境的影响值得深入研究.本文采用OECD标准滤纸接触法、人工土壤法及自然土壤法研究了PFOS对蚯蚓急性致死作用及回避行为的影响.结果表明： PFOS对蚯蚓的急性毒性作用与染毒时间和染毒浓度相关,试验求得滤纸法48 h、人工土壤法14 d和自然土壤法14 d的LC₅₀值分别为13.64 μg·cm^-2、955.28 mg·kg^-1和542.08 mg·kg^-1;人工土壤和自然土壤中蚯蚓在PFOS的最大试验浓度组160 mg·kg^-1中均表现出显著的回避行为,表明蚯蚓可以明显感知较高浓度PFOS污染土壤并作出回避反应.与急性毒性试验的测试终点LC₅₀相比,蚯蚓行为测试终点对PFOS的反应更为敏感.自然土壤中PFOS对蚯蚓的急性毒性大于人工土壤,相同浓度PFOS作用下,蚯蚓于自然土壤中的回避行为较人工土壤明显.     

单甲脒农药对蚯蚓生长影响的实验

本文以单脒农药模拟喷淋实验观察了该农药对蚯蚓生长的影响。结果表明,原药稀释１００倍的高浓度对蚯蚓生有干扰作用,５０倍浓度可抑制蚯蚓生长。     

敌敌畏、毒斯乐单一及复合污染对蚯蚓的急性毒性试验

蚯蚓作为土壤无脊椎动物的代表,在评价土壤生态系统的污染方面起着重要作用.而敌敌畏和毒斯乐都是目前农业生产中使用相当广泛的农药,在土壤、环境中的残留问题一直是研究的热点.本试验主要用两种方法(滤纸接触法和自然土壤法)研究了敌敌畏、毒斯乐及其混剂对自然蚯蚓的急性毒性.结果 显示:用滤纸接触法测得敌敌畏对蚯蚓的LC50为0.14 g/L,毒斯乐对蚯蚓的LC50为0.154 g/L,敌敌畏和毒斯乐混合药剂对蚯蚓24 h 的LC50为0.102 g/L,而用自然土壤法测得敌敌畏对蚯蚓的LC50为1.11 g/L,毒斯乐对蚯蚓的LC50为1.29 g/L,敌敌畏和毒斯乐混合药剂对蚯蚓的LC50为0.964 g/L.     

阿维菌素B1a对土壤微生物和蚯蚓的影响

研究了不同阿维菌素B_1a浓度对种土壤微生物生长和呼吸强度的影响及对土壤中蚯蚓的急性毒性.结果表明,土壤中阿维菌素B_1a浓度在83.3 mg·kg^-1以上时对种土壤细菌均表现出明显的抑制作用,但对土壤真菌不表现抑制作用.高浓度阿维菌素B_1a对土壤微生物的呼吸强度有抑制作用,并且在不同土壤其作用有差异.采用滤纸接触试验和人工土壤试验测定阿维菌素对蚯蚓的急性毒性(半数致死量,LD₅₀),接触毒性LD₅₀为.63μg·cm^-2,土壤法试验测定的LD₅₀在处理后第7 d和第1 d分别为2.13和17.06 mg·kg^-1.     

农药对家蚕(Bombyx mori L.)的亚致死效应研究进展

明确农药对家蚕的影响对综合评价农药的生态效应与合理使用农药具有理论和实践意义.从生长发育、经济性状和食物利用方面综述了农药对家蚕的亚致死效应;从生理代谢、组织器官亚显微结构和培养细胞方面介绍了亚致死效应产生的机制,并结合杀虫剂对其它鳞翅目昆虫亚致死影响方面的研究结果对其进行了讨论,提出了今后加强研究的重点.总的来说,亚致死剂量农药的胁迫可使家蚕幼虫眠性不齐,眠蚕体重和蛹重减轻,发育历期延长,严重者结茧(上蔟)时间推迟,且吐丝营茧功能失常,最终导致蚕茧质量下降.桑叶上残留的农药在一定程度上还会对家蚕的取食行为以及食物摄入后的消化吸收产生影响.农药对家蚕亚致死效应产生的机制目前主要有3个:家蚕体内正常生理水平和代谢平衡受到扰动、家蚕的一些重要组织器官如中肠和后丝腺等微形态结构受到损伤和家蚕卵巢细胞出现凋亡现象.农药对家蚕的亚致死效应因农药种类、试验剂量和家蚕受药时间而异,产生的机理也十分复杂.今后应着重开展新农药对家蚕亚致死效应研究,特别是对生化和分子机制方面的研究以及农药复合污染对家蚕影响的研究.     

三种农药对斑马鱼的急性毒性和生物浓缩系数

采用斑马鱼(Brachydanio rerio)为试验生物,研究了吡虫啉、三唑磷和哒螨灵的急性毒性和生物浓缩系数．通过毒性试验,获得了这3种农药对斑马鱼的24、48、72和96h LC50值．这3种农药的96h LC50值分别为281．37mg·L^-1、8．37mg·L^-1和11．66μg·L^-1．根据急性毒性数据和有关技术指南,确定生物浓缩试验浓度和持续暴露时间为14d．经0．5和5mg·L^-1吡虫啉暴露后的生物浓缩系数(BCFs)分别为1．52和0．97．经0．02和0．2mg·L^-1三唑磷暴露后的BCFs分别为9．00和8．45．经0．1和1μg·L^-1哒螨灵暴露后的BCFs分别为5600和4920．     

蚯蚓提取物对家蚕中肠氧化损伤、总抗氧化能力及抗氧化酶含量的影响

【目的】研究蚯蚓提取物对家蚕Bombyx mori中肠组织氧化应激及抗氧化酶含量的影响,探究蚯蚓提取物的抗氧化功能。【方法】分别给家蚕5龄幼虫饲喂蚯蚓(赤子爱胜蚓Eisenia foetida)提取液原液的50, 100和200倍稀释液处理后的桑叶,测定处理6 d后家蚕中肠组织中氧化损伤产物丙二醛(MDA)和乳酸脱氢酶(LDH)的含量;利用qRT-PCR检测家蚕中肠组织中抗氧化酶关键基因(Cat,Sod和GSH-Px)的mRNA表达水平;同时采用ELISA法检测中肠组织中抗氧化酶[过氧化氢酶(CAT)、超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-Px)]含量及总抗氧化能力(T-AOC)。【结果】与对照组相比,50, 100和200倍稀释组家蚕中肠组织中MDA, CAT, SOD和GSH-Px含量及T-AOC均差异不显著,200倍稀释组LDH含量显著降低了9.85%,100倍稀释组中肠中Cat和GSH-Px的表达量均显著高于对照组,3个稀释组中Sod的表达量均显著高于对照组。【结论】添食蚯蚓提取物可通过降低家蚕中肠LDH含量、提高抗氧化酶基因的表达水平进而提高家蚕的抗氧化能力。     

6种重金属对赤子爱胜蚓的急性毒性效应与风险评价

【背景】近年来,土壤重金属污染问题日益凸显,对生态环境、食品安全和人体健康构成了严重威胁,其急性毒性尚未明确。【方法】采用滤纸接触法和人工土壤法测定了铜(Cu2+)、锌(Zn2+)、镍(Ni+)、镉(Cd2+)、铅(Pb2+)和锰(Mn2+)等6种重金属对赤子爱胜蚓的急性毒性效应,并参照欧盟指令91/414/EEC标准评价了其环境风险。【结果】滤纸接触法48h测定结果表明,6种重金属对蚯蚓的LC50为3.17(2.53～3.81)～90.42(69.45～140.47)μg.cm-2,其毒性次序为Cu2+>Zn2+>Ni+>Cd2+>Pb2+>Mn2+。人工土壤法14d测定结果表明,6种重金属对蚯蚓的LC50为1347(1236～1453)～6936(6144～8930)mg.kg-1,其毒性次序为Cu2+>Cd2+>Ni+>Zn2+>Mn2+>Pb2+。风险评价结果显示,Cu2+、Zn2+、Ni+、Cd2+、Pb2+和Mn2+等6种重金属的暴露比(toxicity/exposureratio,TER)分别为3.37、4.46、8.68、1428、13.87和5.85。【结论与意义】6种重金属对土壤动物蚯蚓均具有潜在的毒性效应。Cu2+、Zn2+、Ni+、Mn2+等4种重金属对赤子爱胜蚓存在急性毒性风险,而Cd2+和Pb2+对赤子爱胜蚓的急性毒性风险水平是可接受的。该评价结果可为我国制定基于风险的土壤环境质量标准提供依据。     

家蚕拟浆细胞具有吞噬功能

目的：通过观察家蚕Bombyx mori吞噬细胞的微细结构,来确定拟绛色细胞是否也具有吞噬功能。方法：用荧光小球微量注射家蚕pnd pS品系的幼虫,经荧光染色剂丫啶橙和碘化丙啶染色循环血细胞后,在荧光显微镜下观察并扫描拍摄。结果：观察发现除颗粒细胞和浆血细胞外,一些原血球细胞（血干细胞）和拟绛色细胞（多酚氧化酶）也能吞噬荧光小球。在拟绛色细胞里还发现许多和颗粒细胞一样的能被丫啶橙染色的颗粒。尽管在小球细胞中没有发现被吞噬的荧光小球,但该类血球有比较多的能被丫啶橙染色的大颗粒,这表明它们可能是已经被吞噬的凋亡小体。结论：除颗粒细胞和浆血细胞外,一些原血球细胞和拟绛色细胞也能吞噬荧光小球。说明拟绛色细胞也具有吞噬功能。     

一种家蚕类浓核病毒的组织病理学特征

本文从家蚕病蚕中分离到一种家蚕类浓核病毒（BmDNV-Like）,对它的组织病理学研究表明:该病毒首先寄生家蚕中肠柱状细胞,继而引起其细胞核的膨大和破裂;组织原位杂交结果表明该病毒既能在家蚕中肠柱状细胞中增殖,也能在中肠的杯形细胞中增殖,甚至在感染后期能在家蚕幼虫的大部分组织细胞中感染和增殖。     

The genome sequence of silkworm, Bombyx mori.

We performed threefold shotgun sequencing of the silkworm (Bombyx mori) genome to obtain a draft sequence and establish a basic resource for comprehensive genome analysis. By using the newly developed RAMEN assembler, the sequence data derived from whole-genome shotgun (WGS) sequencing were assembled into 49,345 scaffolds that span a total length of 514 Mb including gaps and 387 Mb without gaps. Because the genome size of the silkworm is estimated to be 530 Mb, almost 97% of the genome has been organized in scaffolds, of which 75% has been sequenced. By carrying out a BLAST search for 50 characteristic Bombyx genes and 11,202 non-redundant expressed sequence tags (ESTs) in a Bombyx EST database against the WGS sequence data, we evaluated the validity of the sequence for elucidating the majority of silkworm genes. Analysis of the WGS data revealed that the silkworm genome contains many repetitive sequences with an average length of <500 bp. These repetitive sequences appear to have been derived from truncated transposons, which are interspersed at 2.5- to 3-kb intervals throughout the genome. This pattern suggests that silkworm may have an active mechanism that promotes removal of transposons from the genome. We also found evidence for insertions of mitochondrial DNA fragments at 9 sites. A search for Bombyx orthologs to Drosophila genes controlling sex determination in the WGS data revealed 11 Bombyx genes and suggested that the sex-determining systems differ profoundly between the two species.     

虫酰肼及其衍生物0593对家蚕的毒性及作用机理

为明确虫酰肼衍生物0593对家蚕Bombyx mori的毒性,本研究采用食下毒叶法测定了虫酰肼及其衍生物0593对家蚕的毒性,观察了亚致死浓度下对家蚕生长发育的影响,并测定了虫酰肼及其衍生物0593对家蚕幼虫体内保护酶的影响,对虫酰肼0593的作用机理进行了初步探索。结果表明：虫酰肼及其衍生物0593对2龄家蚕96 h的LC₅₀值分别为1.2863和0.3364 mg·L^-1,属高毒级药剂;虫酰肼及其衍生物0593在亚致死剂量下对家蚕的生长发育有明显的不利性,可使幼虫历期缩短0.5～2 d;处理组眠期体重、全茧量、蛹重和化蛹率与对照相比均显著降低;对4龄幼虫体内多酚氧化酶和几丁质酶也有较明显影响,虫酰肼及其衍生物0593处理家蚕,处理后6 h对体内多酚氧化酶有明显激活作用,12 h后表现出显著的抑制作用;虫酰肼及其衍生物0593对家蚕体内几丁质酶均有激活作用。0593对保护酶的影响较虫酰肼明显。结果提示虫酰肼及其衍生物0593对家蚕毒性高,对其生长发育和保护酶类均有不利性,不适合在桑园及其周边农田使用。     

Spindle-E is essential for gametogenesis in the silkworm,Bombyx mori

As a defense mechanism against transposable elements, the PIWI-interacting RNA (piRNA) pathway maintains genomic integrity and ensures proper gametogenesis in gonads. Numerous factors are orchestrated to ensure normal operation of the piRNA pathway. Spindle-E (Spn-E) gene was one of the first genes shown to participate in the piRNA pathway. In this study, we performed functional analysis of Spn-E in the model lepidopteran insect, Bombyx mori. Unlike the germline-specific expression pattern observed in Drosophila and mouse, BmSpn-E was ubiquitously expressed in all tissues tested, and it was highly expressed in gonads. Immunofluorescent staining showed that BmSpn-E was localized in both germ cells and somatic cells in ovary and was expressed in spermatocytes in testis. We used a binary transgenic CRISPR/Cas9 system to construct BmSpn-E mutants. Loss of BmSpn-E expression caused derepression of transposons in gonads. We also found that mutant gonads were much smaller than wild-type gonads and that the number of germ cells was considerably lower in mutant gonads. Quantitative real-time PCR analysis and TUNEL staining revealed that apoptosis was greatly enhanced in mutant gonads. Further, we found that the BmSpn-E mutation impacted gonadal development and gametogenesis at the early larval stage. In summary, our data provided the first evidence that BmSpn-E plays vital roles in gonadal development and gametogenesis in B. mori.     

Extrachromosomal transmission of microinjected DNA to the progeny of silkworm Bombyx mori

Vector DNA (pBmFRT) microinjected into the silkworm eggs (preblastoderm stage) persisted in different conformational forms throughout the period of embryonic development. Southern blot analysis confirmed the persistence of DNA as extrachromosomal copies. Slot blot analysis showed the inheritance of the injected DNA to the subsequent progenies; however the copy number of the injected vector declined in the progenies.     

Functional analysis of four Gloverin-like genes in the silkworm, Bombyx mori

To identify genes involved in the innate immunity of the silkworm Bombyx mori, we constructed a cDNA library from the fat body of Escherichia coli-challenged B. mori larvae. Based on the expressed sequence tag (EST) data and whole genome shotgun sequence analysis, we found four Gloverin-like genes, BmGlov1-4, in the Bombyx genome. Northern blot and RT-PCR analysis showed that BmGlov1-4 were induced in the larval fat body after an immune challenge by the injection of E. coli; however, less induction was observed after the injection of a yeast Candida albicans. In silico sequence analysis revealed the presence of a motif homologous to NF-kappaB binding site in the upstream region of each BmGlov gene. Moreover, we expressed recombinant BmGlov1-4 proteins using the baculovirus expression system, and found that all the recombinant BmGlov1-4 significantly inhibited the growth of E. coli.     

Purification and characterization of cocoonase from the silkworm Bombyx mori

Cocoonase (CCN) which facilitates the degradation of a cocoon is recognized as a trypsin-like serine protease. In this study, CCN from the silkworm Bombyx mori was purified and comprehensively characterized. Its activity was maximal at about pH 9.8. It was stable above pH 3.4 at 4?°C and below 50?°C at pH 7.5. CuSO₄, FeSO₄, and ZnSO₄ showed inhibitory effects on CCN, but other salts improved activity. Typical trypsin inhibitors inhibited CCN, but the relative inhibitory activities were much lower than those against bovine trypsin. An extract of cocoon shells inhibited trypsin, but it was only slightly inhibitory against CCN. There were significant differences in catalytic efficiencies and substrate specificities as between CCN and bovine trypsin.