桑天牛成虫交配后血淋巴和生殖系统内可溶性总糖和蛋白质含量的变化

以生理状况相似的未交配桑天牛Apriona germari Hope雌、雄成虫为供试昆虫, 观察、记录交配活动;采集血淋巴,解剖生殖系统并绘图;用蒽酮比色法和福林-酚法检测交配前、后成虫血淋巴和生殖系统内可溶性总糖和蛋白质含量变化。结果表明：交配后1 h,桑天牛雄虫血淋巴内的可溶性总糖含量增加21.38%,蛋白质含量降低22.66%;雄虫生殖系统内的可溶性总糖和蛋白质含量都明显升高;雄性附腺作为某些特异性蛋白质的合成场所其内的可溶性总糖和蛋白质含量分别降低81.76%和63.76%,雌虫血淋巴和卵巢内的可溶性总糖和蛋白质含量都升高。交配后, 雄虫发生陪伴行为最短历时为4 h, 可能是其重要的生殖策略之一。     

淡足侧沟茧蜂寄生对甜菜夜蛾幼虫血淋巴总糖、蛋白质及虫体脂质含量的影响

为探讨淡足侧沟茧蜂Microplitis pallidipes Szepligeti调控寄主的生理机制,测定了淡足侧沟茧蜂寄生后,甜菜夜蛾Spodoptera exigua(Hübner)幼虫血淋巴总糖、蛋白质及虫体脂类含量的变化。研究结果表明,寄生后连续5 d的观察时间内,甜菜夜蛾幼虫血淋巴总糖含量从寄生后第1天开始就高于未寄生寄主幼虫,且在寄生后第2至5天达到显著水平;除寄生后第3天外,被寄生寄主幼虫血淋巴总蛋白质含量始终低于未寄生幼虫,且在寄生后第1、4、5天达到显著水平;甜菜夜蛾被寄生后,虫体脂质含量始终高于未寄生幼虫,且从寄生后第2天开始达到显著水平。本研究揭示,淡足侧沟茧蜂寄生刺激了寄主甜菜夜蛾幼虫血淋巴总糖合成,但抑制了其蛋白质合成,同时也刺激了甜菜夜蛾虫体脂质合成。     

管氏肿腿蜂寄生及注射其毒液对黄粉甲蛹营养代谢的影响

本文以肿腿蜂科管氏肿腿蜂Scleroderma guani为研究对象,研究该蜂寄生及毒液对寄主黄粉甲Tenebrio molitor蛹营养代谢的影响。通过Braford法、香兰素-浓硫酸法、蒽酮法和3’5-二硝基水杨酸法分别测定管氏肿腿蜂寄生及注射其毒液对黄粉甲蛹血淋巴和脂肪体中蛋白质、脂类和糖类含量的影响。研究发现管氏肿腿蜂寄生和注射其毒液能引起黄粉甲蛹血淋巴中的蛋白质含量增加和脂类含量下降,而脂肪体中的蛋白质含量减少和脂类含量升高。同时,该蜂寄生和注射其毒液,均能引起黄粉甲蛹血淋巴和脂肪体中的总糖、海藻糖和还原糖含量升高,但糖原含量下降。研究结果表明,管氏肿腿蜂毒液能调控寄主黄粉甲蛹血淋巴和脂肪体中营养物质的代谢,为揭示该蜂对寄主的营养代谢调控机制奠定了基础。     

微孢子虫和狄斯瓦螨分别侵染后的意蜂血淋巴蛋白质含量变化

微孢子虫Nosema apis和狄斯瓦螨微孢子虫Nosema apis和狄斯瓦螨 Varroa destructor (Acari: Varroidae)均为危害意蜂Apis mellifera的重要寄生虫,该文对其危害后意蜂血淋巴蛋白质含量的变化进行了研究。用考马斯亮蓝法测定了意蜂受侵染后血淋巴的蛋白质总量,并用高压超薄层等电点聚焦法进行血淋巴蛋白质分类。结果显示,病蜂血淋巴蛋白质总量,在人工感染微孢子虫后1～10天呈微孢子虫Nosema apis和狄斯瓦螨 Varroa destructor (Acari: Varroidae)均为危害意蜂Apis mellifera的重要寄生虫,该文对其危害后意蜂血淋巴蛋白质含量的变化进行了研究。用考马斯亮蓝法测定了意蜂受侵染后血淋巴的蛋白质总量,并用高压超薄层等电点聚焦法进行血淋巴蛋白质分类。结果显示,病蜂血淋巴蛋白质总量,在人工感染微孢子虫后1～10天呈上升趋势,然后逐渐下降,感染后12～27天保持在感染前意蜂血淋巴总蛋白质含量水平以下。螨侵染后意蜂血淋巴蛋白质含量明显增高,与健康意蜂相比差异极显著。高压超薄层等电点聚焦分析表明：狄斯瓦螨自然侵染意蜂后,意蜂血淋巴蛋白质组分与健康对照组相比发生了明显改变。这些结果提示,意蜂对于微孢子虫或狄斯瓦螨的侵染产生了一定的免疫反应。     

前裂长管茧蜂寄生行为对桔小实蝇幼虫生理指标的影响

通过观察测定了桔小实蝇幼虫生长发育过程中血淋巴蛋白种类和血细胞的变化以及前裂长管茧蜂的寄生行为对桔小实蝇幼虫各项生理指标的影响。结果表明:不同日龄桔小实蝇幼虫的血细胞浓度随着虫龄的增加呈显著上升趋势,由2龄的16.53×106cells/mL,到3龄的30.14×106cells/mL直至蛹前期的35.94×106cells/mL,但血淋巴蛋白种类没有明显变化。和未寄生幼虫相比,寄生后的幼虫各类型血细胞的浓度均下降,但差异均不显著;血淋巴蛋白种类无明显增减,但浓度有所变化;寄生4 h后血淋巴蛋白质浓度显著降低,接近22 h时升高,至化蛹前期浓度再次下降;寄生行为使幼虫的发育历期从8 d延长至9～11d;4日龄幼虫在被寄生后的第4 d起体重显著高于未被寄生的桔小实蝇幼虫。     

华北大黑鳃金龟幼虫血细胞的超微结构观察

应用光学和电子显微镜技术检查了华北大黑鳃金龟3龄幼虫血淋巴内的血细胞,识别出5种类型的血细胞(原血胞、浆血胞、颗粒血胞、珠血胞和凝血胞)并对每一种血细胞的超微结构特点进行描述。     

椰扁甲啮小蜂寄生对椰心叶甲蛹免疫反应的影响

为了测明椰扁甲啮小蜂Tetrastichus brontispae寄生对寄主椰心叶甲Brontispa longissima蛹的血细胞和体液免疫反应的影响,开展了椰扁甲啮小蜂寄生对椰心叶甲蛹血细胞数量和延展性、血淋巴酚氧化酶活性、血淋巴黑化百分率和血细胞凝集素活性等影响的研究。结果表明：与同期未被寄生蛹相比,寄生蛹血细胞总量在寄生后2 d显著降低,但寄生后4 d显著升高; 寄生蛹的浆血细胞延展率在寄生后2 d显著降低,寄生后4 d显著升高;寄生蛹的血淋巴黑化百分率在寄生后0.5～2 d较高,寄生后3～4 d降低直至为0;寄生蛹的血淋巴酚氧化酶活性在寄生后0.5 d,1 d和4 d时显著升高;寄生蛹的血凝素活性在寄生后2 d较高,寄生后1 d和4 d较低。结果说明椰扁甲啮小蜂寄生使寄主椰心叶甲蛹血细胞和体液免疫反应呈现不规律的变化。     

光周期诱导甘蔗花芽分化及成花逆转过程中生理代谢的变化

以两个在自然条件下难开花的甘蔗品种‘ROCl6’和‘ROC22’为材料,研究经光周期诱导甘蔗花芽分化和成花逆转过程中叶片碳水化合物和可溶性蛋白质含量的变化。结果表明,未分化前,两品种经光周期诱导后叶片可溶性总糖、淀粉和蛋白质含量与对照相比无明显变化;从花芽分化诱变期开始,两品种叶片可溶性总糖、淀粉和蛋白质含量迅速增加,明显高于对照（未分化）。随着花芽进一步分化,其叶片可溶性总糖、淀粉含量逐渐降低并低于对照,处在一个较低的水平,蛋白质含量始终高于对照。成花逆转植株在花芽分化前期叶片可溶性总糖、淀粉和蛋白质含量也高于对照,但低于正常抽穗开花植株;之后随着花芽分化的停止,其叶片可溶性总糖、淀粉含量一直保持较稳定的水平,仅蛋白质略有降低。说明甘蔗花芽分化期间大量的基因表达并且合成各种蛋白质的过程需要消耗大量的碳水化合物。     

胚后发育期臭腹腺蝗中的血细胞种类

应用血球计数器统计了胚后发育期臭腹腺蝗Zonocerus variegatus中存在的血细胞类型和数目。从1龄幼虫至成虫的发育阶段中共观察到6种血细胞类型,即原血细胞 （PRS）、 浆血细胞 （PLS）、粒细胞 （GRS）、珠血细胞 （SPS）、绛色细胞（OES） 和adipohaemocytes （ADS）。不过,在1龄幼虫期未发现OES。在这6种血细胞中,PLS的总平均数最高,OES的总平均数最低。成虫期的血细胞数目显著高于其他发育阶段（P<0.05）,而1龄幼虫和2龄幼虫期的血细胞数目不存在显著差异（P>0.05）。     

昆虫差异蛋白质组: 进展和展望

黑腹果蝇Drosophila melanogaster、家蚕Bombyx mori、意大利蜜蜂Apis mellifera和冈比亚按蚊Anopheles gambiae等昆虫的基因组测序已经基本完成,蛋白质组学技术将是阐明这些基因组功能的重要工具。本文综述了应用差异蛋白质组学技术在昆虫诱导性免疫、抗性机制和分子病理研究方面取得的一些成果：（1） 细菌、真菌、脂多糖或机械损伤诱导的昆虫血淋巴或血细胞来源细胞系的蛋白质表达的改变; （2） Bt抗性昆虫中肠刷状缘膜和抗锥虫采采蝇Glossina morsitans唾液腺多种蛋白质表达的改变; （3）化学农药处理和姬蜂Chelonus inanitus携带的多分DNA病毒引起的昆虫蛋白质组的变化。最后讨论了昆虫差异蛋白质组学研究的瓶颈与对策。     

The effect of nucleopolyhedrovirus infection and/or parasitism by Microplitis pallidipes on hemolymph proteins, sugars, and lipids in Spodoptera exigua larvae

We determined the physiological effects of joint and separate nucleopolyhedrovirus (NPV) infection and parasitism by the endoparasitoid Microplitis pallidipes Szepligeti on biochemical events in the noctuid Spodoptera exigua (Hübner). The results indicated that in parasitized larvae, compared to healthy larvae, total protein concentration in host hemolymph began to decline and total sugar concentration significantly increased by the first day, while lipid content in the host body significantly increased by the second day after parasitization. Meanwhile, in jointly infected and parasitized hosts, compared to parasitized larvae, total protein concentration was consistently higher, total sugar concentration was consistently lower, and lipid content became higher by the second day after treatment. In virus-infected larvae, compared to healthy larvae, total protein concentration sharply declined during the first two days but increased by the third, while total sugar concentration increased on the second and third days after virus infection but decreased at other observation times, and lipid content began to increase by the second day after virus infection. Finally, in larvae that were both parasitized and virus-infected, compared to just virus-infected larvae, total protein concentration increased during the first two days but decreased by the third, total sugar concentration increased only on the first and fourth days, and lipid content decreased significantly on the first day but began to increase by the second day after treatment. These findings led us to conclude that parasitization inhibited protein mobilization but stimulated sugar mobilization in host hemolymph, and promoted lipid mobilization in the host body, while Spodoptera exigua NPV infection stimulated protein mobilization induced by parasitization but inhibited sugar mobilization induced by parasitization.     

The effect of abscisic acid on sugar levels in seedlings of Phaseolus vulgaris L. cv. redland pioneer

Abscisic acid (ABA) caused an increase in total-sugar and a 3-fold increase in reducing-sugar content in the roots of intact bean seedlings. The level of reducing sugars was also increased in the stem but total sugar levels remained unaffected by ABA. ABA also increased reducing-sugar content of the root in seedlings with cotyledons removed but in this case the reducing- and total-sugar contents of the leaf were reduced. However, ABA did not affect reducing- and total-sugar levels in excised bean root systems. The observed increase in sugar content of the root of intact bean seedlings appears to be the consequence of an ABA-induced stimulation of sugar transport from the shoot to the root. It is proposed that a relationship may exist between the effect of ABA on sugar transport and its effect on ion transport in excised root systems and in intact seedlings.     

Total protein in crawfish hemolymph as a parameter of functional state of animals and biomarker of quality of habitat

Analysis of total protein in hemolymph was performed by Lowry method on sexually mature crawfish Pontastacus leptodactylus. The total protein content in the crawfish hemolymph was studied for one year. The protein concentration varied widely, amounted to from 12 to 95 mg/ml, and depended on season and the moulting cycle phase. There are presented histograms for distribution of animals for the protein level for different seasons and their character is analyzed. In summer the amount of protein is maximal prior to moult and decreases by 40 % at once after it. There is studied the diapason of total protein concentrations in hemolymph, in which survival of crawfish at unfavorable changes in habitat is maximal. The adaptive possibilities of crawfish with the low protein content are reduced. The crawfish with the protein concentration in hemolymph lower than the "critical" one were submitted for different time by action of hydroquinone (1 g/l) used as a model toxicant. A brief action did not affect the protein content in hemolymph. At a long toxic action the protein level in hemolymph fell, on average, by 40%, which preceded the death of the animals. Possible mechanisms of positive correlation of the protein concentration in the crawfish hemolymph and of their survival at deterioration of quality of the water medium are discussed.     

Changes of acid phosphatase content and activity in the fat body and the hemolymph of the flesh fly Neobellieria (sarcophaga) bullata during metamorphosis

Changes in the specific and total activity of the lysosomal marker enzyme acid phosphatase (Acph) and in the amount of enzyme protein were examined in the fat body and the hemolymph from the last larval molt to the larval-pupal apolysis. The specific activity showed minor changes during the last larval period. In contrast, the total activity of the enzyme was low during the feeding period and higher during the wandering stage and strikingly increased at the time of puparium formation. We purified a protein having para-nitrophenyl phosphate phosphatase (Acph) activity and raised antisera against it. The amount of Acph protein in the fat body and hemolymph was examined using an ELISA. The specific Acph content showed little variation, but the total amount of the enzyme protein showed a stepwise increase in both organs during last larval stage and was markedly elevated in the pupal stage in the fat body. In contrast, a considerable decrease in the amount of Acph protein was observed in the hemolymph during this period. These data were in agreement with immunohistochemical observations showing an accumulation of the enzyme protein in fat body cells during the prepupal stage with a concomitant disappearance of the enzyme from the hemolymph. Inhibition of ecdysteroid secretion by water stress prevented the changes both in total enzyme activity and in the amount of Acph protein. However, Acph protein content and enzyme activity could be restored when the water stress was followed by a 20-hydroxyecdysone (20-HE) treatment. Taken together, our data show that Acph is secreted by fat body cells into the hemolymph during the larval stage, where it is stored in an inactive form. Increase in the 20-HE titer at the end of last larval stage reverses this process, and the enzyme is taken up by the fat body cells, where it becomes activated and appears in auto- and heterophagic vacuoles. Arch. Insect Biochem. Physiol. 34:369–390, 1997. © 1997 Wiley-Liss, Inc.     

Total protein in hemolymph of crawfish <Emphasis Type="Italic">Pontastacus leptodactylus</Emphasis> as a parameter of the functional state of animals and a biomarker of quality of habitat

Analysis of total protein in hemolymph was performed by Lowry method on sexually mature crawfish Pontastacus leptodactylus. The total protein content in the crawfish hemolymph was studied for one year. The protein concentration varied widely, amounted to from 12 to 95 mg/ml, and depended on season and the molting cycle phase. There are presented histograms for distribution of animals for the protein level during different seasons and their character is analyzed. In summer the amount of protein is maximal prior to molt and decreases by 40% at once after it. There is studied the diapason of total protein concentrations in hemolymph, in which survival of crawfish at unfavorable changes in habitat is maximal. The adaptive possibilities of crawfish with the low protein content are attenuated. The crawfish with the protein concentration in hemolymph lower than the «critical» one were submitted for different time by action of hydroquinone (1 g/l) used as a model toxicant. A brief action did not affect the protein content in hemolymph. At a long toxic action the protein level in hemolymph fell, on average, by 40%, which preceded death of the animals. Possible mechanisms of positive correlation of the protein concentration in the crawfish hemolymph and of their survival at deterioration of quality of the water medium are discussed.     

Hemolymph of Anopheles stephensi from noninfected and Plasmodium berghei-infected mosquitoes. 3. Carbohydrates.

Determinations were made of carbohydrates in hemolymph collected from adult female mosquitoes (Anopheles stephensi). First the hemolymph was fractionated by extraction and precipitation procedures, after which qualitative and quantitative determinations of carbohydrates were made by thin layer chromatography. The most abundant sugars found in the hemolymph were glucose and trehalose, though maltose, glucuronic acid, and inositol could be found after the mosquitoes took blood meals. After the mosquitoes ingested a noninfected blood meal, their hemolymph sugar levels rose almost 4-fold. There was less of an increase following a blood meal infected with the rodent malaria parasite, Plasmodium berghei. Depletion of sugars in the hemolymph of infected mosquitoes may result from direct utilization of sugar by the malaria parasite developing within the mosquito.     

Characterization of a bacterial agglutinin in the hemolymph of the hard clam, Mercenaria mercenaria

The hemolymph of the hard clam, Mercenaria mercenaria, was found to agglutinate nonspecifically 4 of the 30 bacteria tested and a marine alga. The agglutinin is a protein (or a conjugated protein) because it is: (1) precipitated by trichloroacetic acid and ammonium sulfate; (2) inactivated by extraction with chloroform, but not with toluene or xylene; and (3) inactivated by chymotrypsin and protease, but not by deoxyribonuclease. Electrophoretic analysis shows that the agglutinin is composed of subunits each with a molecular weight of approximately 21,000. Calcium ions are required for the activity of the agglutinin and contribute to the heat stability of the molecule. Several saccharides, which may constitute a portion of the bacterial agglutinin receptors, were capable of partially inhibiting agglutination. In vitro studies using clam hemocytes showed that the phagocytosis of a marine bacterium, designated as RS-005, was enhanced by the presence of hemolymph. Adsorption of hemolymph samples with RS-005 bacteria removed the agglutinin activity for all types of cells tested and also abolished the opsonic effect.     

Correlation of hemocyte counts with different developmental parameters during the last larval instar of the tobacco hornworm, Manduca sexta

We determined the changes in hemocyte titer and in the abundance of hemocyte types of the tobacco hornworm Manduca sexta during the fourth and fifth larval stadium and the beginning of the pupal stadium. As we analyzed the samples of individual insects at daily intervals, we were able to correlate phenotypical features, body weight, as well as total protein content and lysozyme activity in the hemolymph with the observations on hemocytes. In the course of the fifth larval stadium, the hemocyte titer decreased slightly and declined further after pupation. Using calculated values for total hemocyte numbers, females had about five times and males three times more hemocytes in the circulating population at the beginning of the wandering stage (in the middle of the fifth larval stadium) than immediately after the last larval--larval molt (from the fourth to the fifth larval stadium). This sexual difference was mainly due to an increase in the number of plasmatocytes, which was more prominent in females than in males. Granular cells were dominant in early fifth larval stadium while plasmatocytes were the most abundant cells in pupae. Oenocytoids and spherule cells disappeared during the wandering stage. Lysozyme activity in the hemolymph rose to a maximum during the wandering stage, with females having lysozyme values twice as high as those for males. These changes in lysozyme activity, however, did not correlate with the increase of total hemolymph protein titer which occurred already at the beginning of the wandering stage. We postulate that changes in hemocyte titers are under direct hormonal control, which has to be proven in future experiments.     

Intraspecific variations in the hemolymph of Biomphalaria glabrata, a snail host of Schistosoma mansoni.

An attempt was made to characterize the hemolymph of Biomphalaria glabrata with reference to "normal" intra-specific variation, i.e., both inter- and intra-strain differences. Total protein concentration, per cent hemoglobin, pH, and osmolarity were studied. Seven geographic strains of B, glabrata were examined. In addition, observations were made on the hemolymph of Biomphalaria straminea, several strains of Helisoma caribaeum, and on B. glabrata subjected to infection with Schistosoma mansoni or to periods of starvation. Intra-strain differences in total protein concentration and total hemoglobin concentration in B. glabrata appeared to be more closely related with snail size than with absolute age. Inter-strain variation in B. glabrata was also noted, but the differences were of the same magnitude as those from intra-strain samples. Significant differences in total protein concentration were observed, however, between the means of similar size B. glabrata, B. straminea and H. caribaeum. The osmolatity of the hemolymph from different size B. glabrata was similar as were the osmolalities of the hemolymph from similar size snails of different strains. However, all B. glabrata strains exhibited hemolymph osmolalities lower than observed in strains of H. caribaeum. Infection with S. mansoni reduced the protein concentration of B. glabrata hemolymph. Differences were noted as early as 1.5-24 hr post-infection, with significant alterations occurring at about 11 days post-infection. To a lesser extent, starvation also depleted the protein content of the hemolymph.