硬蜱中蜕皮激素含量的变化(英语)

为阐明蜕皮激素在硬蜱发育和生殖中的作用,用高效液相色谱法(HPLC)和放射免疫测定法测定了银盾革蜱和长角血蜱中饱血和产卵前后雌虫整体、血淋巴、合神经节及卵巢中蜕皮激素含量的变化.此外,还用HPLC测定了长角血蜱的卵和幼虫中蜕皮激素的含量.结果表明:在饱血前雌虫整体中蜕皮激素含量的变化不大,饱血后迅速增加.血淋巴中的蜕皮激素在饱血后第5天(即产卵前1天)达到高峰,可做为产卵的信号.合神经节中蜕皮激素含量的高峰在饱血后第5天,与血淋巴中高峰出现的时间一致,这与神经分泌细胞的分泌活性变化相吻合.从饱血后第3天起,卵巢中蜕皮激素含量增加很快,直到产卵时止,在长角血蜱的卵中也测到α-蜕皮素,来源于卵巢,随胚胎发育进程其含量逐渐增加.卵产出后第4天到第12天,蜕皮激素的含量增加十几倍.     

入侵害虫蔗扁蛾成虫及卵内蜕皮激素的定性定量分析

利用液相色谱和放射免疫分析法首次对入侵害虫蔗扁蛾（Opogona sacchari Bojer）成虫及卵内的蜕皮激素进行了定性和定量分析,以期明确蔗扁蛾成虫及卵内蜕皮激素的主要组分及动态变化规律．羽化后第4天,雄成虫蜕皮激素含量仅为0．080ng／只,而雌成虫蜕皮激素含量高达5．978ng／只,差异达到极显著水平．鉴于雌成虫的蜕皮激素主要是由卵巢合成分泌,因此我们测定了卵巢发育过程中整个蜕皮激素的变化动态,发现前期卵巢蜕皮激素含量较低,后期则相对较高,其峰值出现在发育到第3天的卵巢,蜕皮激素含量为10．480ng／卵巢．卵内蜕皮激素含量测定表明,前3天的卵内蜕皮激素含量相对稳定,维持在0．010ng/卵左右,而到第4天时,卵内蜕皮激素含量则下降到0．006ng／卵．蜕皮激素定性分析发现,卵巢和卵内均含有3种主要的蜕皮激素组分：20-羟基蜕皮酮、26-羟基蜕皮酮和一个尚未鉴定的组分．     

长角血若虫发育期20—羟基皮酮含量变化与表皮发生的关系

用高效液相色谱和放射免疫分析对长角血蜱Haemaphysalis longicornis若虫发育期整体蜕皮激素（20－E）含量变化进行了测定,并用扫描电镜对其表皮结构及其发生进行了观察。20－E存在于菲虫整修发育期,饥饿、吸血和饱血后前4天,激素水平低（0.71－1.30ng/头）;饱血后6天明显增加,此时皮溶离开始,蜕皮间隙形成;饱血后7天继续增加;饱血后8天,20－E含量达到高峰（8.70ng/头）,此时若虫开始沉积新的上表皮;高峰后,20－E含量急剧下降,直到蜕皮前保持低水平,此期内沉积新的原表皮,旧表皮被部分吸收,上述结果显示出20－E含量急剧增加及其高峰分别与启动皮层溶离和新的上表皮沉积相吻合,20－E与新的原表皮沉积无关。     

褐飞虱卵巢发育及其与迁飞的关系

描述了褐飞虱雌虫卵巢的构造和发育过程中的变化。根据其形态特征,划分为五个发育级。 环境条件对长翅型雌虫卵巢发育有明显的影响。在高温、低温或取食黄熟期水稻时,卵巢发育停滞;光照期对印巢发育无明显影响。证实了揭飞虱雌虫卵巢的发育是受体内保幼激素所控制。 1977年在我国南方稻区五个点上系统解剖捕获的褐飞虱雌虫,观察了卵巢在迁飞过程中的发育状态。在虫源迁出地区,由于水稻黄熟和温度的影响,田间雌虫卵巢出现生殖滞育现象。在迁飞运行中受高空低温和停止取食的影响,卵巢仍处于滞育状态。在迁入区降落后,温度、食料适宜,解除了生殖滞育,卵巢继续发育而定居繁殖。 提出了褐飞虱卵巢发育与迁飞过程关系的模式图。     

银盾革蜱若虫发育期20-羟基蜕皮酮含量变化规律(英语)

本文利用高效液相色谱(HPLC)对银盾革蜱饥饿、吸血和饱血后若虫整体水平20-羟基蜕皮酮含量变化规律进行了研究.结果表示20-羟基蜕皮酮存在于若虫整个发育期,其含量变化具有两个峰.第一个峰小,出现在吸血期(吸血后5天),含量为2.34ng/只,该峰出现后,若虫快速饱血并从宿主体落下;第二个峰大,出现在蜕化期(饱血后8天),含量为16.17ng/只,此峰可能与上表皮沉积有关.从饱血后第6天至饱血后第8天,20-羟基蜕皮酮含量迅速增加,皮层溶离发生在饱血后7天左右.     

伊维菌素对长角血蜱生长发育及生殖的影响

【目的】通过分析宿主皮下注射伊维菌素对长角血蜱生长发育及生殖的影响,进而评价伊维菌素对长角血蜱的防治效果。【方法】以新西兰白兔为宿主,皮下注射不同剂量伊维菌素,观察幼蜱、若蜱及雌蜱的叮咬率、死亡率、吸血期、饱血体重、产卵量和孵化率等生物学参数,分析伊维菌素对长角血蜱生殖和生长发育的影响。【结果】伊维菌素对长角血蜱各期蜱虫的叮咬率均无显著影响。与对照组相比,伊维菌素可显著增加幼蜱和若蜱死亡率,延长若蜱和雌蜱的吸血期并降低其饱血体重,降低雌蜱产卵量及卵孵化率,抑制卵巢及卵的发育。【结论】宿主皮下注射伊维菌素能够显著抑制长角血蜱的发育及生殖,可作为防治长角血蜱的候选杀虫剂。     

长角血蜱若虫发育期20——羟基蜕皮酮含量变化与表皮发生的关系

用高效液相色谱和放射免疫分析对长角血蜱Haemaphysalislongicornis若虫发育期整体蜕皮激素(20-E)含量变化进行了测定,并用扫描电镜对其表皮结构及其发生进行了观察.20-E存在于若虫整个发育期,饥饿、吸血和饱血后前4天,激素水平低(0.71～1.30ng/头);饱血后6天明显增加,此时皮层溶离开始,蜕皮间隙形成;饱血后7天继续增加;饱血后8天,20-E含量达到高峰(8.70ng/头),此时若虫开始沉积新的上表皮;高峰后,20-E含量急剧下降,直到蜕皮前保持低水平,此期内沉积新的原表皮,旧表皮被部分吸收.上述结果显示出20-E含量急剧增加及其高峰分别与启动皮层溶离和新的上表皮沉积相吻合,20-E与新的原表皮沉积无关.     

银盾革蜱的中肠上皮变化与血餐消化

用光学显微镜及电子显微镜对不同生理状况下银盾革蜱Dermacentor niveus Neumann雌虫中肠上皮及血餐消化进行了研究.饿蜱的中肠只由一种干细胞组成,脂滴作为饿蜱营养的贮藏形式.非滞育蜱消化分三个阶段,即第一连续消化阶段,减慢消化阶段及第二连续消化阶段.吸血后中肠上皮共观察到四种细胞类型,即替代细胞、分泌细胞、消化细胞及卵赞原细胞.滞育蜱第一连续消化阶段延长.饱血后60天到120天,消化作用几近停止,为停滞消化阶段.卵黄原细胞的超微结构有明显改变.滞育解除后,开始进行减慢消化阶段及第二连续消化阶段.     

金泽革蜱的生物学特性研究

金泽革蜱是分布于东洋区的三寄主蜱, 成虫活跃于夏季到晚秋, 其发育历期随季节而不同.雌虫在兔休上吸血时间, 七、八月为16—20天, 十一月只需9天.产卵前期在夏季饱食的雌虫为13—27天;十一月饱食者产生滞育, 至少需244天.产卵期持续31—43天(七、八月), 总产卵量为1749—8995粒.雌虫产卵显与饱血雌虫体重之间有非常显著的正相关(r=0.989, p<0.001).金泽革蜱的产卵力(饱食后每毫克体重产卵数)为7.631.卵期为55—79天(九、十月), 幼虫在兔体上吸血4—6天, 饱食后经18—29天脱皮为若虫, 若虫寿命可达124天.     

昆虫病原线虫斯氏线虫和异小杆线虫对长角血蜱雌蜱的致病力

用昆虫病原线虫小卷蛾斯氏线虫（Sc BJ）、夜蛾斯氏线虫（Sf Otio）、拟双角斯氏线虫（Sc D43）、格氏斯氏线虫（Sg NC32）和嗜菌异小杆线虫（Hb E-6-7）对长角血蜱雌蜱进行感染试验,所用线虫剂量为4 000 Ijs/dish。结果表明,5种线虫均对长角血蜱雌蜱有致死效应。Hb E-6-7和Sc BJ两种线虫对雌蜱各发育期致病力最强,导致雌蜱的累积死亡率和半致死时间分别为饥饿雌蜱82.5%,9.0天和75.0%,8.8天;吸血雌蜱90.0%,8.0天和82.5%,8.0天;饱血雌蜱93.3%,7.3天和86.7%,7.3天。Sc D43对饱血雌蜱有较高的致死效应,为80.0%,但半致死时间较长,为11.7天。Sf Otio和Sg NC32对长角血蜱雌蜱的致死效应较低。饱血雌蜱较饥饿雌蜱和吸血雌蜱更易被线虫感染。     

THE ROLE OF ECDYSTEROIDS IN THE REPRODUCTIVE DIAPAUSE OF DERMACENTOR NIVEUS NEUMANN

Abstract  The ecdysteroid levels in hemolymph, ovary, synganglion and whole body of diapausing female Dermacentor niveus were detected by HPLC, and compared with the results of nondiapausing female. It is revealed that the ecdysteroid levels in hemolymph and ovary of diapausing female are similar basically to that of nondiapausing female in the first few days after engorgement. From the 10th day after engorgement, the ecdysteroid levels of diapausing female decreased and even became distinctly lower than that of nondiapausing female. The paucity of ecdysteroids in these individuals would influence the normal development of oocytes. In order to explore the effect of ecdyateroids on the diapausing female, we injected 20-hydroxyecdysone with different dosages at different time into the ticks, and found that after just complete engorgement the injection with large dosages (10000 and 1375 ng/tick) caused death of the ticks. From 10th to 20th day after engorgement the ecdysteroid levels of diapausing female are lower than that of nondiapausing one before oviposition, the injection with certain dosages 50, 70 and 100 ng/tick> of 20E can accelerate vitellogenesis and terminate reproductive diapause, but the amount of eggs produced by them is less than that produced by nondiapausing female. The termination of diapause in female of ixcdid tick by exogenous ecdysteroids is reported for the first time.     

QUANTITATIVE VARIATION OF ECDYSTEROIDS OF IXODID TICKS

Abstract  In order to explore the role of ecdysteroids in development and reproduction of ixodid ticks, we studied the quantitative variation of ecdysteroids in the hemolymph, synganglion, ovary and whole body of the female ixodid ticks, Dermacentor niveus and Haemaphysalis longicornis , before and after engorgement and oviposition by HPLC and RIA. The ecdysteroid content in eggs of these ticks was determined by HPLC. The results indicated that before engorgement the quantitative variation of ecdysteroids in the whole female body was not significant, but their levels increased rapidly after engorgement. The ecdysteroid titer in hemolymph was peaked on the 5th day after engorgement, which was one day prior to oviposition. It may be regarded as a singnal of oviposition. In the synganglion the peak of ecdysteroid level occurred also on the 5th day after engorgement. This is coincident with the secretory activity of neurosecretory cells of synganglion. From the 3rd day after engorgement until oviposition the ecdysteroid level in the ovary increased rapidly. Ecdysteroids were detected in eggs of H. longicornis too. They stem from ovary and accumulated with the process of embryonic development.     

鞭角华扁叶蜂蜕皮甾类激素滴度的变化

用放射免疫分析法测定了鞭角华扁叶蜂Chinolyda flagellicornis末龄幼虫及滞育预蛹血淋巴中蜕皮甾类激素滴度。结果表明,末龄幼虫血淋巴中蜕皮甾类激素滴度在第2和4天各有一高峰;滞育预蛹血淋巴中保持一定滴度的蜕皮甾类激素,并随发育时期的不同有所波动;预蛹化蛹前一周血淋巴中蜕皮甾类激素滴度存在两个与变态相对应的峰值。表明鞭角华扁叶蜂的滞育与蜕皮甾类激素相关。     

Haemolymph ecdysteroid titres in diapause and nondiapause larvae of the European corn borer, Ostrinia nubilalis

The ecdysteroid titres of last-instar prediapausing, diapausing and nondiapausing larvae of Ostrinia nubilalis were determined by radioimmunoassay. In the nondiapause larvae a major peak of ecdysteroid activity preceded pupation by 24 h and continued through the pupal ecdysis. This peak was correlated with head and thorax critical periods as well as with changes in behaviour and physiology marking the transition from feeding larva to prepupa. Nondiapause larvae also displayed a rise in ecdysteroid titre during the feeding phase of development. This rise was approx one tenth that of the major peak and lasted 32 h. It was not correlated with any overt changes in larval physiology or behaviour. The diapause ecdysteroid profile was distinctive in that the levels measured were all lower than the lowest of the nondiapause curve. No peaks were observed in the diapause titres. Prepupal changes such as spinning and the cessation of feeding were not correlated with any increase in ecdysteroid levels. During diapause termination, under a long-day photoperiod, no increases in ecdysteroid titre were observed for the first 10 days. After 12 days individuals began to show ecdysteroids above the diapause levels. Pupation started after 16 long days.     

滞育和非滞育棉铃虫血淋巴类固醇蜕皮素含量变化的比较

采用放射免疫分析法对不同时期的注定滞育和非滞育棉铃虫的血淋巴中的类固醇蜕皮素的含量进行了测定,发现在预蛹期间,注定滞育的棉铃虫的类固醇蜕皮素含量高于非滞育的棉铃虫,化蛹后,注定滞育的棉铃虫的类固醇蜕皮素含量则迅速降到极低的水平,明显低于非滞育棉铃虫。用20-羟基蜕皮素处理不同时期的滞育蛹,均能打破滞育。由此可见,类固醇蜕皮素含量的降低或缺乏乃是导致棉铃虫滞育的关键因子之一。     

A possible role of 20-hydroxyecdysone in embryonic development of the silkworm Bombyx mori

It has been well established that eggs of insects, including those of the silkworm Bombyx mori, contain various ecdysteroids and the amounts of these ecdysteroids fluctuate during embryonic development. In order to know the function of egg ecdysteroids in embryonic development of B. mori, we examined the biological activities of various egg ecdysteroids by in vitro ligand-binding assay and bioassay using B. mori eggs. First, using the ecdysteroid receptor of B. mori (BmEcR-B1/BmUSP heterodimer) prepared by yeast and Escherichia coli expression systems, the interaction between the ecdysteroid receptor and various egg ecdysteroids of B. mori was analyzed. The relative binding affinities of egg ecdysteroids to the BmEcR-B1/BmUSP heterodimer decreased in the order of 20-hydroxyecdysone > 2-deoxy-20-hydroxyecdysone > 22-deoxy-20-hydroxyecdysone > ecdysone > 2-deoxyecdysone > ecdysone 22-phosphate. Next, several egg ecdysteroids of B. mori were injected into the prospective diapause eggs, which show a very low level of free ecdysteroids at the onset of embryonic diapause (gastrula stage). Approximately 7% of them (P < 0.002, chi(2)-test) developed beyond the gastrula stage without entering diapause by the injection of 20-hydroxyecdysone (25 ng/egg). In contrast, the injection of other ecdysteroids was not effective in inducing embryonic development. These results suggest that 20-hydroxyecdysone, via the ecdysteroid receptor, is responsible for the developmental difference between diapause and non-diapause in B. mori embryos. Furthermore, it was suggested that continuous supply of 20-hydroxyecdysone may be required to induce embryonic development.     

Ovarian Ecdysteroidogenesis in Both Immature and Mature Stages of an Acari,Ornithodoros moubata

Ecdysteroidogenesis is essential for arthropod development and reproduction. Although the importance of ecdysteroids has been demonstrated, there is little information on the sites and enzymes for synthesis of ecdysteroids from Chelicerates. Ecdysteroid functions have been well studied in the soft tick Ornithodoros moubata, making this species an excellent candidate for elucidating ecdysteroidogenesis in Chelicerates. Results showed that O. moubata has at least two ecdysteroidogenic enzymes, Spook (OmSpo) and Shade (OmShd). RNAi showed both enzymes were required for ecdysteroidogenesis. Enzymatic assays demonstrated OmShd has the conserved functions of ecdysone 20-hydroxylase. OmSpo showed specific expression in the ovaries of final nymphal and adult stages, indicating O. moubata utilizes the ovary as an ecdysteroidogenic tissue instead of specific tissues as seen in other arthropods. On the other hand, OmShd expression was observed in various tissues including the midgut, indicating functional ecdysteroids can be produced in these tissues. In nymphal stages, expression of both OmSpo and OmShd peaked before molting corresponding with high ecdysteroid titers in the hemolymph. In fed adult females, OmSpo expression peaked at 8–10 days after engorgement, while OmShd expression peaked immediately after engorgement. Mated females showed more frequent surges of OmShd than virgin females. These results indicate that the regulation of synthesis of ecdysteroids differs in nymphs and adult females, and mating modifies adult female ecdysteroidogenesis. This is the first report to focus on synthesis of ecdysteroids in ticks and provides essential knowledge for understanding the evolution of ecdysteroidogenesis in arthropods.     

Anteroposterior gradient during nymphal-adult moulting cycle of the tropical bont tick,Amblyomma variegatum (Acarina: Ixodidae)

Summary The structure of the extensible (alloscutum) and inextensible (scutum) integument of the nymph, Amblyomma variegatum was examined during the whole bloodmeal and the nymphal-adult moulting cycle. Integumental events were tentatively correlated with the ecdysteroid levels measured by radioimmunoassay. We observed that all the integumental events were realised along an anteroposterior gradient. During the 5 days corresponding to the bloodmeal, although the hormone concentration was low, a new endocuticle was deposited on both the alloscutum and scutum. Furthermore, mitoses were initiated in the capitulum. On days 1–2 after the meal, ecdysteroid titres began to increase and reached a first peak corresponding to 4.1 ng 20-hydroxyecdysone equivalents/tick on the 4th day after the ticks dropped off their host. At this time the epidermis of the capitulum was detached and the outline of the adult capitulum was already visible. Mitotic activity in the alloscutum was initiated. On day 6 post-drop, the frontal apolysis was achieved and the ecdysteroid titres declined to basal values. A second peak much higher than the first one (maximum value of 33.7 ng/tick) and identified principally as 20-hydroxyecdysone by HPLC/RIA was noted on the 13th day post-drop. During the period of increase in the ecdysteroid levels (days 9–10 post-drop), the mitotic phase ended in the alloscutum and the apolysis began. Epicuticle was deposited after day 12 postdrop. Then, while the titre fell to low values (about 1.6 ng/tick, days 16–20 post-drop), the exocuticle was deposited and the nymphal cuticle was digested. All adult structures were functional 3 days before ecdysis. In young male as in female adults the mean value of the ecdysteroid levels corresponded to about 2.5 ng/tick. Finally, hydrolysis of tick whole extracts with esterase demonstrated a low increase of RIA-positive material, demonstrating the probable presence of natural ecdysteroid fatty-acid conjugates in this species.     

The hormonal regulation of the larval diapause in the codling moth, Laspeyresia pomonella (Lep. Tortricidae)

The hormonal control of the facultative diapause of the codling moth has been investigated. The diapause can be divided into 4 phases or periods: (1) diapause induction by short-day conditions (SD) in young larvae, (2) initiation of the diapause in the early last larval instar by a high titre of juvenile hormone, (3) onset and maintenance of diapause with inactivity of the neuroendocrine system, as evidenced by the results of neck-ligation experiments, (4)termination of diapause by the production of ecdysteroid.Diapause-induced larvae pupated after spinning the cocoon, if the state of induction was changed by injection with the anti-juvenile hormone precocene II at the beginning of the last larval instar and subsequent results of neck-ligation experiments, (4) termination of diapause by the production of ecdysteroid. treated with juvenile hormone during the first 1.5 days after the last larval moult and subsequently reared under SD. Under LD, continuous application of juvenile hormone during the last larval instar and after spinning did not prevent the insects from moulting to either a supernumerary larva, a pupa or a larval-pupal intermediate. Termination of diapause, i.e. pupation, was achieved by injecting diapausing larvae with 20-hydroxyecdysone. Although juvenile hormone was found to have a prothoractropic effect in diapausing larvae, no pupal moult could be induced by the application of the hormone. Contrary to the hormonal situation before pupation of nondiapausing larvae, no juvenile hormone could be detected before or during the pupation of larvae after diapause.