用ELISA方法研究稻田节肢动物的食物关系

应用酶联免疫吸附试验双抗体夹心法,研究了稻田节肢动物,包括19种捕食性天敌、4种主要水稻害虫及1种中性昆虫之间的食物关系。在检测的19种捕食者中,有15种捕食了白背飞虱,11种捕食了褐飞虱。其中食虫沟瘤蛛、拟水狼蛛、八斑鞘蛛、粽管巢蛛、四斑锯螯蛛和拟环纹豹蛛对两种稻飞虱的捕食阳性率较高。有7种捕食了稻纵卷叶螟幼虫,其中粽管巢蛛、褶管巢蛛、拟环纹豹蛛、青翅蚁形隐翅虫和印度细颈步甲的捕食阳性率较高。拟环纹豹蛛、浙江豹蛛和拟水狼蛛对稻蝗若虫有较高的阳性率。13种捕食者捕食了摇蚊,摇蚊作为替代或补充猎物,对稻田捕食性节肢动物亚群落的重建和发展具有重要意义。在实验检测的基础上,构建了被检测节肢动物猎物与捕食者间的食物关系图。     

间斑寇蛛 L.tredecimguttatus粗毒抗血清制备及其活性分析

间斑寇蛛Latrodectus tredecimguttatus是世界上毒性最强的蜘蛛之一,其毒液中含有许多大分子物质,可用于制备抗血清。将粗毒过滤除菌后作为抗原,加弗氏佐剂背部皮下免疫豚鼠。加强免疫3次后经检测抗血清效价达到1：8,再次加强免疫后断颈处死豚鼠放血,分离血清,经饱和硫酸铵分级沉淀及阴离子交换柱层析纯化后,采用westernblot法检测抗体所在峰。离体大鼠输精管电刺激收缩反应实验及活体动物给药实验均表明抗体对毒素具有明显的中和作用。特异性间斑寇蛛毒素抗血清为治疗间斑寇蛛咬伤奠定了基础。     

群特异性蓝舌病病毒单克隆抗体的制备和鉴定

目的：制备群特异性抗蓝舌病病毒（BTV）单克隆抗体,并对其特性进行鉴定,为建立检测BTV抗原及抗体的ELISA方法奠定基础。方法：用纯化的BTV颗粒为免疫抗原免疫BALB/c鼠,以大肠杆菌表达的VP7蛋白作为筛选抗原,用间接ELISA法筛选杂交瘤细胞株;选取抗体效价最高的一株制备BTV单克隆抗体,以该抗体为捕获抗体与8种不同血清型BTV进行ELISA反应,结果与细胞病变反应进行比对;以该抗体为竞争抗体,与12种不同血清型绵羊BTV抗血清进行竞争ELISA反应,并将结果与参比c-ELISA试剂盒结果进行比对。结果：筛选出5株稳定分泌BTV单克隆抗体的杂交瘤细胞株,并选其中一株（3E2）制备了高纯度的单克隆抗体;该单抗用于检测不同血清型BTV,与细胞病变反应结果完全相符;用于检测不同血清型绵羊BTV抗血清,其结果与参比c-ELISA试剂盒符合率为100%,与鹿流行性出血热病毒抗原和抗体均无交叉反应。结论：制备的BTV单克隆抗体具有良好的群特异性,可用于检测不同血清型BTV抗原及BTV抗体。     

三突花蛛对小菜蛾幼虫的捕食效应

本文研究三突花蛛对小菜蛾幼虫的捕食功能反应及干扰因素对捕食作用的影响。结果表明,三突花蛛对小菜蛾幼虫的捕食效应属HollingⅡ型反应,捕食作用率随捕食者-猎物比率上升而下降,雌蛛的捕食效应强于雄蛛。     

三突花蛛对小菜蛾幼虫的捕食效应

本文研究三突花蛛对小菜蛾幼虫的捕食功能反应及干扰因素对捕食作用的影响。结果表明,三突花蛛对小菜蛾幼诉捕食效应属ＨｏｌｌｉｎｇⅡ型反应,捕食作用率随捕食者－猪物比率上升而下降,雌蛛的捕食效应强于雄蛛。     

拟环纹狼蛛对褐飞虱的捕食作用及其模拟模型的研究Ⅲ.选择捕食作用

本文研究了拟环纹狼蛛雌成蛛对褐飞虱、稻纵卷叶螟的选择捕食作用。在不同猎物类型共存和不同总猎物密度下,测定了捕食者对猎物的喜好性和转换行为,分析了捕食者对猎物的功能反应形式,喜好性和转换行为与共存猎物种类、数量之间的关系;建立了在多种猎物类型共存时,雌成蛛对猎物的总捕食作用方程及对每一种猎物类型的捕食作用方程。室内验证实验表明:所建立的捕食作用方程具有一定的描述能力。     

嗜肺巴氏杆菌外膜蛋白和脂多糖抗原在血清学诊断中的意义

目的评价嗜肺巴氏杆菌外膜蛋白(OMP)和脂多糖(LPs)作为血清学诊断抗原的敏感性和特异性.方法用OMP、LPS和全菌(WC)作为Western blot和ELISA的诊断抗原检测自然感染和实验感染嗜肺巴氏杆菌小鼠相应的IgG抗体滴度,同时测定3种抗原与实验动物常见致病菌的交叉反应.结果与嗜肺巴氏杆菌自然感染和实验感染小鼠血清的ELISA反应中,不同时期,LPS作为诊断抗原时血清抗体阳性率最高,WC次之,OMP最低.自然感染小鼠群中,出生4周LPS抗体阳性率即可达80%,而同期的WC和OMP仅为25%和20%,故LPS敏感性最高.与实验动物常见致病菌免疫血清和阴性种鼠血清的ELISA反应中,WC抗原表现出较高的吸光度(A)值,经Western blot证实,其反应为非特异性反应,LPS抗原特异性最强,OMP抗原次之.结论混合多株具有型或种特异性的OMP或LPS作为ELISA的诊断抗原,无论从特异性和敏感性上均高于全菌抗原.     

草间小黑蛛对茶蚜的捕食功能反应

在实验室条件下研究了草间小黑蛛(Erigonidium graminicolum)对茶蚜(Toxoptera aurantii)的捕食作用。草间小黑蛛对茶蚜的捕食功能反应属于HollingⅡ型。草间小黑蛛有较强的种内干扰反应,随着捕食者密度的增大,草间小黑蛛的捕食率相应降低。猎物密度和天敌密度相互干扰会降低草间小黑蛛的寻找效应,但对捕食量没有影响。     

肿瘤患者血清中SARS-CoV抗体阳性原因分析

探讨SARS冠状病毒(SARS—CoV)抗体在SARS病原学诊断中的特异性及其在肿瘤患血清中的假阳性问题。应用ELISA和荧光定量RT-PCR技术检测了111例正常对照和40例肿瘤患血清中SARS—CoV抗体的阳性率。在111例正常对照中,IgM抗体均阴性,IgG抗体的阳性率为3．6％(4／111);IgG抗体诊断SARS的特异性为96．4％,两种抗体同时阳性诊断SARS的特异性为100％。40例肿瘤患中,IgM抗体均阴性,IgG抗体阳性率17．5％(7／40)。经RT—PCR检测,上述肿瘤患阳性病例均为阴性。结果表明,同时测定SARS—CoV的两种抗体可降低诊断的假阳性率,提高诊断的特异性。用非纯化SARS—CoV抗原制备的ELISA试剂盒测定肿瘤患的SARS—CoV抗体,可能出现假阳性。在肿瘤患中出现假阳性的原因可能与包被的抗原有关。     

温度变化对休耕期冬水田天敌捕食猎物的影响及天敌耐冷性分析

为评估冬水田休耕季节长达6-8个月温度变化对稻田主要捕食性天敌通过觅食策略维持其种群发生的影响,在室内测定了两种代表性温度下青翅蚁形隐翅甲Paederus fuscipes成虫、拟水狼蛛Pirata subpiraticus和拟环纹豹蛛Pardosa pseudoannulata成蛛、亚成蛛对天台刺齿虫兆Homidia tiantaiensis和白翅叶蝉Thaia rubiginosa成虫的捕食功能反应、搜寻效应、自身密度干扰效应以及捕食偏好性;测定了这些捕食者在这两种温度诱导下的过冷却点（Supercooling point,SCP）和结冰点（Freezing point,FP）。结果表明,温度变化显著影响了捕食性天敌的捕食功能反应、对猎物的搜寻效应以及自身密度的干扰反应。在22℃和12℃下,捕食者对刺齿虫兆和叶蝉的捕食功能反应均符合Holling II模型。由捕食功能反应可知,22℃下捕食者对两种猎物的捕食量均高于12℃下的捕食量,两种温度下捕食者对刺齿虫兆的捕食量要显著高于对叶蝉的捕食量。22℃下捕食者对两种猎物的瞬时攻击率、最大捕食量和a/T_h值均相应地高于12℃下的值,而22℃下它们对两种猎物的处理时间则相应地小于12℃下的处理时间。在同一猎物相同温度下,狼蛛和豹蛛亚成蛛的瞬时攻击率、最大捕食量和a/T_h值均相应地高于狼蛛和豹蛛成蛛的值,它们的处理时间则明显小于其成蛛的处理时间。隐翅甲的最大捕食量和a/T_h值在12℃叶蝉试验中均最大,而其处理时间则最小;其它条件下的隐翅甲参数值通常介于狼蛛和豹蛛成蛛、亚成蛛各参数值之间。捕食者对两种猎物的搜寻效应与猎物密度呈负相关;温度变化对捕食者搜寻效应的影响程度与捕食者和猎物的种类有关;22℃下的搜寻效应值均高于12℃下的搜寻效应值。捕食者对两种猎物的捕食作用均随自身密度升高而下降,22℃下对两种猎物和12℃下对刺齿虫兆的搜寻常数和干扰系数值均以狼蛛和豹蛛亚成蛛的为最高,而12℃下对叶蝉的这2个值以豹蛛亚成蛛和隐翅甲的为最高。两种温度下捕食者对这两种猎物存在选择偏好性,更嗜食刺齿虫兆,但温度变化没有对捕食者的这种猎物偏好性产生影响。在这两种温度快速冷驯化下这些捕食者间的SCP和FP均差异显著,通常温度降低能诱导亚成蛛的SCP和FP下降,而成蛛的升高,隐翅甲的变化很小或下降,说明不同捕食者适应温度降低的耐冷特性与其捕食作用间存在一定相关性。     

Laboratory infection of blackflies (Simuliidae) and midges (Chironomidae) by the mosquito mermithid, Romanomermis culicivorax

Penetrating infective juveniles of Romanomermis culicivorax usually killed first-stage larvae of Chironomus maturus Johann., Chironomus sp., Simulium damnosum Theo., and S. venustum Say. Nematodes were melanized and died after they entered fourth stage larvae of 2 chironomid species, but no host reaction was evident after entry into fourth-stage blackfly larvae. In contrast, the nematodes initiated development in the latter hosts, which died before the nematodes completed their development.     

用ELISA方法定量评价食虫沟瘤蛛对稻飞虱的捕食作用(英文)

对捕食性节肢动物的捕食作用进行评价是害虫生物防治研究的一个重要内容。本文利用酶联免疫吸附试验（ＥＬＩＳＡ）定量评价了食虫沟瘤蛛对稻飞虱的捕食作用。结果表明,方法特异性完全符合试验要求;食虫沟瘤蛛在捕食３头３－５龄白背飞虱或褐飞虱若虫后的检出期分别为９６小时和１２０小时;在早稻田中,捕食性天敌对白背飞虱和褐飞虱的阳性反应率分别为１９．０５％－４７．３４％和１９．０５％－６６．６７％,在飞虱密度较低时,捕食性天敌仍表现出较高的阳性率;捕食量随飞虱密度的增加而增加,但捕食率下降。捕食性节肢动物是调节褐飞虱种群动态的重要因子之一。     

QUANTIFYING PREDATION BY UMMELIATA INSEC-TICEPS BOES. ET STR. (ARANEAE: LINYPHIIDAE) ON RICE PLANTHOPPERS USING ELISA*

Abstract An enzyme-linked immunosorbent assay (ELISA) has been developed to quantify predation by Ummeliata insecticeps Boes. et Str. on rice planthoppers in paddy fields in Dasha Township, Guangdong Province. The assay was completely specific for rice planthopper materials. The detection periods for antigens after an adult of U. insecticeps had fed on three 3–5th instar nymphs of white-back planthopper (WBPH), Sogatella furcifera (Horvath), and brown planthopper (BPH), Nilaparvata lugens (Stal), at room temperature were 96 h and 120 h, respectively. The proportion of individual species of predators scoring positive ranged from 19. 05% to 47. 34% for WBPH, and from 9. 25% to 66. 67% for BPH in the early rice season. Although the numbers of the rice planthoppers consumed by U. insecticeps increased with increasing prey densities, the predation rates declined. When rice planthopper densities were low, the predation rates were relatively high. This kind of predation can explain why the rice planthoppers have never reached outbreak levels in rice fields in Dasha Township since 1973.     

中国双叶摇蚊亚属记述(双翅目,摇蚊科,摇蚊属)(英文)

对中国摇蚊属的双叶摇蚊亚属作了系统学记述。描述了2个新种:宽铗双叶摇蚊Chironomus(Camptochironomus)latusus sp.nov.和六毛双叶摇蚊Chironomus(Camptochironomus) senisetosus sp.nov.,重新描述了苍白双叶摇蚊Chironomus(Camptochironomus) pallidivittatus和伸展双叶摇蚊Chironomus(Camptochironomus) tentans。并编制了中国本亚属4种雄成虫分种检索表。宽铗双叶摇蚊,新种Chironomus(Camptochironomus) latusus sp.nov.(图1～2)雄虫生殖节构造与苍白双叶摇蚊相似,但本种肛尖两侧叶强烈骨化,抱器端节明显宽大。AR值明显小于后者(表2)。正模♂,内蒙古乌梁素海,1982-04-25,王新华灯诱。六毛双叶摇蚊,新种Chironomus(Camptochironomus) senisetosus sp.nov.(图5～6)雄虫生殖节构造与伸展双叶摇蚊相似,但可借以下特征区别于后者(表4):上附器发达,6根长刚毛有序地围绕并着生在上附器基部,肛节侧片毛缺失,腋瓣缘明显少于后者。正模♂,贵州省罗甸县,2004-08-11,于昕网捕。     

广东双季稻区杂草地和稻田中捕食性节肢动物的群落动态

用吸虫器采样法,于1998年对广东省大沙镇双季稻区两块相邻的杂草地和稻田中捕食性节肢动物群落的结构进行了研究。在杂草地生境中共采集到73种捕食性节肢动物,其中55种为蜘蛛,18种为昆虫。3月21日,早稻田翻耕前,在杂草地中采集到33种捕食性节肢动物,密度为130头/m²。4月4日,水稻移栽1周后,在杂草地中采到29种捕食性节肢动物,密度为92头/m²;同期在稻田中采到12种捕食性节肢动物,密度为16.2头/m2。5月13日早稻成熟前期,在杂草地中只采到19种捕食性节肢动物,密度为28头/m²;而此期,在稻田中采到27种捕食性节肢动物,密度为53.2头/m²。在晚稻生长期,杂草地捕食性节肢动物与稻田捕食性节肢动物物种数和密度的变化与早稻生长期情况相似。杂草地与稻田两生境间的捕食性节肢动物群落的相似性系数大于0.5,由此可见,这两类生境中捕食性节肢动物的物种组成是非常相似的,具有较多的共有种。杂草地捕食性节肢动物群落可能是稻田捕食性节肢动物群落重建的重要种库之一。     

Exposure of arthropod predators to Cry1Ab toxin in Bt maize fields

Abstract.  1. To assess the risks of an insect-resistant transgenic plant for non-target arthropods, it is important to investigate the exposure of non-target species to the transgene product. Exposure of predators in the field depends on the toxin levels in food sources, their feeding ecology and that of their prey.
2. To verify the transmission of Cry1Ab toxin through the food chain, and thus exposure of predators in the field, samples from different plant tissues, herbivores, and predators in Bt maize fields in Spain (Event 176) were collected at different periods over the season and the toxin content was measured using ELISA. Complementary laboratory studies were performed with the omnivorous predator Orius majusculus to assess the toxin uptake and persistence after feeding on variable Bt-containing food sources.
3. Field results revealed that toxin content in some herbivores was negligible (aphids, thrips, leafhoppers) compared with those in spider mites. The latter herbivore only occurred after pollen shed and contained three times greater toxin levels than Bt maize leaves.
4. Data confirmed that the Bt toxin can be transferred to predators, that is to say to Orius spp., Chrysoperla spp., and Stethorus sp. This only applied when Bt maize pollen or spider mites were available. The passage of Bt toxin to O. majusculus via these two food sources was also confirmed in the laboratory. Contrastingly, some predators in the field (hemerobiids, Nabis sp., Hippodamia sp., Demetrias sp.) contained no or negligible toxin levels even when pollen or spider mites were present.
5. Besides essential information for exposure assessment of numerous arthropod predators, this study provides an insight into the feeding ecology of different arthropods in the maize system.     

Prey location and prey choice by the freshwater leech Erpobdella octoculata using foraging kairomones

1. The freshwater leech, Erpobdella octoculata , is a generalist predator feeding on prey organisms such as Tubifex spp., Chironomus spp. and Asellus aquaticus. Using different experimental designs, we studied the use of foraging kairomones by the leeches for prey location.
2. Leeches were attracted to living as well as to freshly killed larvae of Chironomus sp., to Tubifex sp., and to A. aquaticus offered in Petri dishes covered with gauze. The leeches also reacted to an extract of macerated Tubifex sp. presented in agar.
3. Using ion exchange chromatography, the presence of histidine and glutamic acid was demonstrated in water contaminated by living larvae of Chironomus sp. and Tubifex sp. Agar blocks containing a synthetic mixture of these compounds at concentrations above 5 mg mL⁻¹ were attractive to the leeches.
4. Choice tests showed that leeches preferred chemical signals from Tubifex sp. over larvae of Chironomus sp. No difference was found between chemicals from Tubifex sp. and A. aquaticus , and A. aquaticus and larvae of Chironomus sp.
5. The results demonstrate that E. octoculata uses specific foraging kairomones in searching for prey and indicate that amino acids serve as foraging kairomones.     

用ELISA方法评价捕食性天敌对白背飞虱的捕食效能

用酶联免疫吸附试验（ELISA）方法测定了9类捕食性节肢动物对白背飞虱的捕食阳性反应率,定量评价了拟水狼蛛对白背飞虱的捕食效能。结果表明：活动能力强,与稻飞虱生境相似的捕食性天敌种类表现出较高的阳性反应率;拟水狼蛛对白背飞虱的捕食效率随水稻的生长而加强,在水稻生长后期,分别为21.53％和43．08％。     

稻田及其毗邻杂草地寄生蜂群落结构与特征

通过在稻田及其毗邻杂草地的系统调查、采集和室内标本鉴定,获知稻田中有67种寄生蜂,隶属16科;毗邻杂草地中有96种寄生蜂,隶属19科。稻田生境中的寄生蜂物种数占稻田天敌总物种数的45．89％,杂草生境中的寄生蜂物种数占杂草地天敌总物种数的46．60％。对稻田及其毗邻杂草地寄生蜂群落的各种属性指标进行比较研究,结果表明：杂草地寄生蜂群落的丰富度、多样性、优势集中度指数均比稻田高,而均匀性指数值比稻田低。对杂草生境和稻田生境寄生蜂群落的优势种动态比较可知,寄生蜂优势种在水稻的不同生育期是不相同的,在稻田生境中,优势种稻虱缨小蜂(Anagrus sp．)出现在水稻生育前期和中后期,等腹黑卵蜂(Telenomus dignus)出现在水稻生育后期,稻苞虫黑卵蜂(Telenomus sp．)在水稻生育前中期及后期出现。然而在杂草地中,作为优势种,稻虱缨小蜂出现在水稻生育前期和中期,等腹黑卵蜂出现在水稻生育前中期和后期,稻苞虫黑卵蜂在水稻整个生育期均有出现。两种锤角细蜂(Monelata sp．1和Acropiesta sp．2)是稻田的重要优势种,但不是杂草地的优势种;相反,稻蝽小黑卵蜂(Telenomus gifuensis)是杂草地的重要优势种,但不是稻田的优势种。由此可见,杂草地生境是稻田寄生蜂的种库之一,对稻田节肢动物群落的重建和种群保存有重要影响,因而适当保留田埂杂草和稻田周围的杂草地,减少对天敌的损害,提高天敌对害虫的自然控制作用,是水稻害虫持续控制的途径之一。