Platelet aggregation and sphingomyelinase D activity of a purified toxin from the venom of loxosceles reclusa

A facile and quantitative assay for measuring the activity of sphingomyelinase D in recluse spider venom has been developed using L-α-[palmitoyl-1-¹⁴C]lysophosphatidylcholine as substrate. This assay avoids the problem of substrate insolubility that occurs when sphingomyelin and other lipids are used as subtrates. This assay has been employed in gel filtration and isoelectric focusing isolation techniques to purify sphingomyelinase D from spider venom. The purified sphingomyelinase exhibits four active enzyme forms in isoelectric focusing with pI values of 8.7, 8.4., 8.2, and 7.8. Each active form when examined in SDS-polyacrylamide gel electrophoresis gave an estimated molecular weight of 32 000. The four active enzyme forms were immunologically cross-reactive with each other as demonstrated with radioimmune assays using an antiserum developed to one of the active forms. Each active form hydrolysed sphingomyelin to release choline and produce N-acylsphingosine phosphate. One of the active enzyme forms was characterized further in dermonecrosis and platelet aggregation measurements. This purified sphingomyelinase D was identified as a poisonous toxin that can develop the typical dermonecrotic spider lesion when injected into experimental animals at levels expected to be delivered in a normal bite. Furthermore, the purified toxin acts to aggregate human blood platelets. The toxin-induced platelet aggregation has been related to serotonin release as aggregation occurs, and it has been shown to be inhibited by EDTA over the range of 0.6 to 3.0 mM EDTA. It is suggested that spider-induced dermonecrosis could result in part from platelet aggregation at and near the site of envenomation.     

Nanoparticle self-assembly by a highly stable recombinant spider wrapping silk protein subunit

Artificial spider silk proteins may form fibers with exceptional strength and elasticity. Wrapping silk, or aciniform silk, is the toughest of the spider silks, and has a very different protein composition than other spider silks. Here, we present the characterization of an aciniform protein (AcSp1) subunit named W₁, consisting of one AcSp1 199 residue repeat unit from Argiope trifasciata. The structural integrity of recombinant W₁ is demonstrated in a variety of buffer conditions and time points. Furthermore, we show that W₁ has a high thermal stability with reversible denaturation at ∼71 °C and forms self-assembled nanoparticle in near-physiological conditions. W₁ therefore represents a highly stable and structurally robust module for protein-based nanoparticle formation.     

Previous experience and site tenacity in the orb spider Nephila (Araneae,Araneidae)

Summary The tenacity of the orb spider Nephila clavipes to a web site was studied in the laboratory. No differences were found between the giving-up-times and the site tenacity of spiders reared in the laboratory or those caught in the field, nor between spiders raised under a poor or a richt diet. The animals left sites at random and seemed to ignore experiences gained at previous sites.     

Relating body size to the role of aquatic subsidies for the riparian spider <Emphasis Type="Italic">Nephila clavata</Emphasis>

We examined the relationship between body size of the riparian spider Nephila clavata and the contribution of allochthonous (aquatic insects) and autochthonous (terrestrial insects) sources to its diet using stable isotope analysis. During the study period from July to September, the body size of the females increased remarkably (about 60-fold) but that of males remained small. The biomass of both aquatic and terrestrial insects trapped on the spider webs increased with spider size, with the biomass of the former ranging between 30 and 70% of that of the terrestrial insects. The average relative contribution of aquatic insects to the diet of the spiders, calculated from δ¹³C values, was 40–50% in spiders in the early juvenile and juvenile stages, 35% in adult males and 4% in adult females. There was a significant negative relationship between the relative contribution of aquatic insects and body size of the female spiders. We conclude that aquatic insects might be an important seasonal dietary subsidy for small spiders and that these allochthonous subsidies may facilitate the growth of riparian spiders, which may in turn enable the spiders to feed on larger prey.     

Amino acid analysis of spider dragline silk using H NMR

The amino acid composition of Nephila clavipes dragline silk fiber was determined by conducting ¹H nuclear magnetic resonance (NMR) spectroscopy experiments on acid-hydrolyzed material. N. clavipes dragline silk was found to consist of 43.0 ± 0.6% Gly, 29.3 ± 0.2% Ala, 9.1 ± 0.1% Glx, 4.0 ± 0.1% Leu, 3.3 ± 0.1% Tyr, 3.4 ± 0.2% Ser, 2.7 ± 0.1% Pro, 2.1 ± 0.1% Arg, 1.07 ± 0.05% Asx, 0.96 ± 0.05% Val, 0.48 ± 0.03% Thr, 0.35 ± 0.03% Phe, and 0.28 ± 0.03% Ile. Compared with standard chromatography-based amino acid analysis (AAA), the chemical resolution of NMR allows for an amino acid solution to be characterized without separation and is shown to provide considerably higher precision. This allows for more accurate statistics on the variability of amino acids in spider dragline silk. In general, this ¹H NMR AAA technique is applicable to a large range of proteins and peptides for precise composition characterization, especially when the precise content of a minor component is critical and relatively large amounts of sample are available (microgram to milligram quantities).     

Fine Structural Analysis of the Central Nervous System in the Spider, Achaearanea tepidariorum (Theridiidae: Araneae)

ABSTRACT Central nervous system (CNS) of arachnids is still mysterious and has a rich unexplored field compare to what is known in insects or crustaceans. The CNS of the spider, Achaearanea tepidariorum, consists of a dorsal brain or supraesophageal ganglion and circumesophageal connectives joining it to the subesophageal mass. As the segmentation of the arachnid brain is still under discussion, we classify the brain as a protocerebral and tritocerebral ganglion depending on the evidences which generally accepted. The subesophageal nerve mass underneath the brain is the foremost part of the ventral nerve cord. All of this nerve mass is totally fused together, and forming subesophageal ganglia in this spider. In the brain, the nerve cells are packed in the frontal, dorsal and lateral areas, but are not absent from the posterior and ventral regions. In addition, the nerve cells of the subesophageal and abdominal ganglia are only restricted to the ventral and ventolateral regions. The CNS of the spider, Achaearanea tepidariorum is similar in feature to the Family Araneidae.     

Biology, life table and predation of Feltiella acarisuga (Diptera: Cecidomyiidae) feeding on Tetranychus cinnabarinus eggs (Acari: Tetranychidae)

Feltiella acarisuga (Vallot) is a common predatory gall midge, which feeds on many species of spider mites. All major life history and life table parameters of F. acarisuga were determined using the carmine spider mite, Tetranychus cinnabarinus (Boisduval) eggs as prey under laboratory conditions [26.7 ± 2 °C, 75 ± 5% RH, and a photoperiod of l4:10 (L:D) h]. Developmental times of F. acarisuga were 2.6, 7.1, and 6.7 d for eggs, larvae, and pupae, respectively, with an average of 16.4 d from egg to adult emergence. Female immatures development took ≈1 d longer than male immatures did. Adult F. acarisuga lived an average of 12.8 d, and the female adults (13.3 d) lived significantly longer than male adults did (11.9 d). After an average of 1.5 d preoviposition period, each female laid an average of 27.3 eggs in its life span with an average of 2.1 eggs per day and an average of 2.8 eggs on each of its oviposition day. The raw data were analyzed using an age-stage, two-sex life table method that takes into consideration of the variable developmental rates among individuals and between sexes. The intrinsic rate of natural population increase (r), net reproductive rates (R_o), gross reproductive rate (∑m_x), generation time (T), and doubling time (DT), and the finite rate of increase (λ) of F. acarisuga were estimated using the age-stage, two-sex (male and female) life table analysis as 0.122 d⁻¹, 16.19 eggs per female, 20.81 eggs per female, 22.81 d, 1.1298 d, and 5.7 d⁻¹, respectively. Each of the first, second, and third instar larvae of F. acarisuga consumed an average of 35.5, 54.0 and 86.9 T. cinnabarinus eggs per day, respectively. Larvae of F. acarisuga could consume an average of 175.4, T. cinnabarinus eggs, and female larvae consumed 14% more spider mite eggs (187.6 eggs) than male larvae (165.1 eggs). The significance of other life table parameters related to the population and the potential of using F. acarisuga as a biological control agent are discussed.     

中国蜘蛛物种多样性分析

基于2011年1月更新的世界蜘蛛名录第11．5版,本文对中国蜘蛛的物种多样性进行了统计和分析：（1）目前中国共记述蜘蛛67科670属3667种,种数占世界蜘蛛的8．72％,仍有许多物种尚待发现;（2）目前中国特有蜘蛛物种共计2376种,占中国蜘蛛种数的64．79％、世界蜘蛛种数（42055种）的5．65％,表明中国蜘蛛特有物种极为丰富,为中国生物多样性的保护、环境及气候的历史演变和蜘蛛的系统演化研究起到重要作用;（3）已知蜘蛛物种数最多的省份为云南（728种,占19．85％）、湖南（612种,占16．69％）、四川（454种,占12．38％）等,一定程度上表明上述各省生物多样性丰富,但这一结果也与各地蜘蛛多样性研究不均衡有关。蜘蛛各科名录及统计数据可参见“中国蜘蛛特有物种网”。     

中国蜘蛛分科检索表

蜘蛛的物种多样性是极其丰富的,但目前只有一小部分的蜘蛛种类被描述。世界上已描述的蜘蛛种类已超过40000种,隶属于110个科。在我们居住的小范围内,可能至少有30个科的数百种蜘蛛。就中国而言,估计可能有40000种以上的蜘蛛种类,但目前也只有大约4000种被命名。本检索表首次列出了中国现有67个蜘蛛科的答定特征．以及不同科之间的相似处和不同处。