The effect of starvation on haemolymph metabolites,fat body and ovarian development in Oxya japonica (Acrididae: Orthoptera)

Haemolymph volume does not change significantly during 96 hr of starvation. Haemolymph and fat body metabolites are depleted significantly in insects denied food for 96 hr. The percentage of oöcyte resorption is increased considerably by starvation. Differences in the utilization of lipid and protein under conditions of starvation between adult Oxya and adult locusts may be attributed to the migratory habit of the latter.     

Effects of residual chlorfluazuron on haemolymph-borne oviposition-stimulating factors in Spodoptera litura

Acylurea insecticides can have various effects on insect reproduction and may even interfere with haemolymph‐borne oviposition‐stimulating factors (OSFs). This study describes the effects of injecting haemolymph into females of the common cutworm, Spodoptera litura (Fabricius) (Lepidoptera: Noctuidae), the effects of chlorfluazuron on the activities of OSFs, when sublethal doses (LD₁₀: 1.00 ng larva^?1 or LD₃₀: 3.75 ng larva^?1) were applied topically to newly ecdysed fifth instars under laboratory conditions. The haemolymph of moths of varying age, sex, and mating status resulted in the following increases in oviposition rates within the first 24 h, listed in decreasing order: the haemolymph of mated females injected into mated females (56%), mated females into newly emerged virgin females (53%), virgin males into virgin females (49%), and virgin females into mated females (29%), compared with uninjected mated or virgin females. Two factors are involved in the activation of ovipositon: OSF‐I is found in virgin females, whereas OSF‐II may be formed or received by males during mating. By contrast, the haemolymph taken from chlorfluazuron‐treated adults (treated as fifth instars with sublethal doses) injected into females resulted in the following percentage decreases in oviposition rates within the first 24 h, listed in decreasing order: the haemolymph of LD₁₀‐ and LD₃₀‐treated virgin males injected into untreated virgin females (77 and 84%, respectively), LD₁₀ and LD₃₀‐treated virgin females into untreated mated females (70 and 80%, respectively), LD₁₀ and LD₃₀‐treated mated females into untreated mated females (61 and 69%, respectively), LD₁₀ and LD₃₀‐treated mated females into untreated virgin females (59 or 68%, respectively), compared with untreated ones. Hence, residual chlorfluazuron decreases the activities of OSFs by significantly decreasing the oviposition rates. Moreover, virgin females’ or males’ OSFs are more sensitive to chlorfluazuron than those of mated cutworms. Both OSFs are most likely proteins.     

FOSSIL BLOOD DROPLETS IN MIOCENE DOMINICAN AMBER YIELD CLUES TO SPEED AND DIRECTION OF RESIN SECRETION

Abstract:  Two spiders (Filistatidae) in Miocene Dominican Republic amber, one newly identified and only the second known fossil of this family, have autospasized legs (detached at a predetermined locus of weakness when restrained by a non-self-induced source) at the patella-tibia joint. In both specimens, droplets of haemolymph (blood) are preserved exiting the patellae. The autospasized legs and the presence of haemolymph suggests that both spiders were engulfed in rapid-flowing resin seeps of relatively low viscosity, rather than having wandered onto a sticky exudate, becoming stuck and then covered by a subsequent resin flow. These are the first reported incidences of such fossilized blood droplets, the shape, size and position of which provide clues to preservational taphonomy, an understanding of which is necessary for reliable conclusions concerning fossil communities and ecosystems. In addition, haemolymph droplets may serve as reservoirs for fossil DNA.     

Nondestructive sampling of insect DNA from defensive secretion

Nondestructive techniques to obtain DNA from organisms can further genetic analyses such as estimating genetic diversity, dispersal and lifetime fitness, without permanently removing individuals from the population or removing body parts. Possible DNA sources for insects include frass, exuviae, and wing and leg clippings. However, these are not feasible approaches for organisms that cannot be removed from their natural environment for long periods or when adverse effects of tissue removal must be avoided. This study evaluated the impacts and efficacy of extracting haemolymph from a defensive secretion to obtain DNA for amplification of microsatellites using a nondestructive technique. A secretion containing haemolymph was obtained from Bolitotherus cornutus (the forked fungus beetle) by perturbation of the defensive gland with a capillary tube. A laboratory experiment demonstrated that the sampling methodology had no impact on mortality, reproductive success or gland expression. To evaluate the quality of DNA obtained in natural samples, haemolymph was collected from 187 individuals in the field and successfully genotyped at nine microsatellite loci for 95.7% of samples. These results indicate that haemolymph-rich defensive secretions contain DNA and can be sampled without negative impacts on the health or fitness of individual insects.     

Detection and localization of antibody ingested with a mosquito bloodmeal

Mouse immunoglobulins were found to persist in Aedes aegypti (L.) mosquito bloodmeals for 2-3 days after ingestion. Immunoenzyme labelling revealed mouse antibody specifically bound to the mosquito midgut epithelium after ingestion; immunogold labelling of thin sections revealed mouse antibody within the cytoplasm of the microvilli on the midgut epithelium. Ingested mouse antibody was not conclusively demonstrated bound to tissues outside the gut, though antibody was detected in mosquito haemolymph using a sensitive ELISA test. Possible mechanisms by which antibody may cross mosquito gut barriers and actions that in vivo antibody:antigen reactions may have on these bloodfed insects are discussed.     

蜜蜂幼虫血淋巴游离氨基酸和微量元素含量的差异对其抗螨特性的影响

重新界定的外寄生螨类---狄斯瓦螨Varroa destructor(Anderson and Trueman),严重危害全世界的西方蜜蜂Apis mellifera。但是对其原始寄主东方蜜蜂Apis cerana不构成可见的危害。在西方蜜蜂群中,狄斯瓦螨在雄蜂房和工蜂房都能进行繁殖。在其亚洲的原始寄主东方蜜蜂群中,它们可以寄生于雄蜂和工蜂,但在工蜂房中不育。蜜蜂的血淋巴是狄斯瓦螨生存和繁殖需要摄取的惟一食物来源,推测血淋巴中的某种物质含量会影响狄斯瓦螨的繁殖。对中华蜜蜂Apis ceranaFabricius和意大利蜜蜂ApismelliferaL.工蜂和雄蜂封盖幼虫血淋巴中游离氨基酸和与营养有关的微量元素含量进行了比较,发现其存在明显差异,并推测这些差异与东方蜜蜂抗螨能力强有关。     

不同家蚕品系的耐热性特征

研究了家蚕Bombyx mori 3个品系(Pure Mysore,NB4D2和CSR2）5龄幼虫和蛹在不同温度（35,38和40℃）下的耐热性,采用Probit分析测定了它们在各温度下的LT50值和置信限。结果表明：多化性品系Pure Mysore在高温下的存活率高于两个二化性品系NB4D2和CSR2,而两个二化性品系中NB4D2表现出更好的耐热性。家蚕幼虫接触38℃高温6 h和40℃高温3 h后,其血淋巴中出现90,70和29 kDa的热激蛋白条带。在恢复过程中,NB4D2和CSR2的血淋巴中未见29 kDa蛋白条带,而Pure Mysore幼虫的血淋巴中29 kDa蛋白仍然表达。当幼虫置于高温下时,血淋巴中90和70 kDa蛋白表达,但是检测不到29 kDa蛋白。研究认为热激蛋白表达与热带家蚕不同品系的耐热性以及与同一品系不同发育阶段的耐热性具有相关性。     

Characterization of lipophorin from Antheraea mylitta drury (Lepidoptera: Saturniidae)

The high molecular weight (680 KDa) glycolipoprotein from the haemolymph of male larvae of Antheraea mylitta Drury (Lepidoptera: Saturniidae) was identified as lipophorin by gradient KBr ultracentrifugation and SDS-PAGE. This lipophorin is composed of two subunits: apolipoprotein 1 (234 KDa) and apoprotein II (80 KDa). The density of the native molecule is 1.1941 g/ml. By weight, it contains 53.7% protein, 3.7% carbohydrate, and 42.6% lipid. Neutral lipids and phospholipids are 66.2 and 33.8% of the total lipids, respectively. Mannose and N-acetylglucosamine are the only sugars detected by gas liquid chromatography. The amino acid composition of both the native molecule and its two subunits was determined, and yielded similar amino acid compositions. © 1996 Wiley-Liss, Inc.     

棉铃虫不同虫态及虫龄血淋巴中酚氧化酶活力的比较

分别测定了棉铃虫Helicoverpa armigeraHübner不同虫态及虫龄血清和血细胞中酚氧化酶(phenoloxidase,PO)的活力。结果显示血清和血细胞中都有酚氧化酶活性,且血细胞中高于血清中。不同虫态及虫龄的血清和血细胞中酚氧化酶活力有很大的不同,血清和血细胞中酚氧化酶活力变化规律一致。3龄幼虫酶活力最高,5龄幼虫最低。酶活力大小依次为:3龄幼虫>预蛹>4龄幼虫>蛹>5龄幼虫     

Metabolomic analyses of the haemolymph of the Asian citrus psyllid Diaphorina citri,the vector of huanglongbing

The Asian citrus psyllid Diaphorina citri Kuwayama is currently threatening the citrus industry by transmitting the causative agent Candidatus Liberibacter asiaticus (CLas) of huanglongbing. Multiplication of CLas in haemolymph of D. citri indicates that it contains the necessary nutrients for CLas. Although many studies examine D. citri, the haemolymph composition of this dangerous pest remains to be investigated. In the present study, the haemolymph of D. citri is collected using a nanolitre syringe after the removal of one of its forelegs. The haemolymph is either derivatized with methyl chloroformate (MCF) or trimethylsilyl (TMS) derivatizing reagent and analyzed with gas chromatography–mass spectrometry. Nineteen amino acids, two organic acids and seven fatty acids are detected in the haemolymph after MCF derivatization. More metabolites are detected after TMS derivatization. Sugars are the most abundant metabolites in the haemolymph. Glucose and fructose are the main monosaccharides. Trehalose and sucrose are the major disaccharides. Furthermore, three inositol isomers (myo‐inositol, scyllo‐inositol and chiro‐inositol) are detected in the haemolymph. Organic acids are found in low amounts, whereas phosphoric acid is found at a higher concentration. Twenty‐four nucleotides and sugar nucleotides, including ATP, ADP and AMP, are detected using high‐performance anion‐exchange chromatography. Adenine nucleotides are the most abundant nucleotides followed by uridine and guanosine. The adenylate energy charge for the haemolymph is 0.77. Our results show that many metabolites found in the citrus phloem sap are also found in the haemolymph of D. citri.