Screening of Necrotizing Spiders in Korea Using Nonradioactive Sphingomyelinase Assay(II) Web-building Spiders

Spider bites cause a range of symptoms from simple swellings to disfiguring necrotic lesions, and occasionally death. While spider bites are not a major medical problem in Korea, it would be of great value to know which species of spiders pose a threat to human health. A middle molecular weight protein, sphingomyelinase D, has been identified in the venom of the brown recluse spider and strong evidence suggests that they have a major role in spider bite necrosis. For the identification of necrotizing species, we have investigated using recently developed non‐radioactive assay of sphingomyelinase for rapidly screening the necrotizing venoms. Here, we demonstrate the fetal toxicity of total 65 species (24 genera, 9 families) of the web‐building spiders among 622 identified spider species in Korea. It has been revealed that four species of the orb‐weaving spider, Araneus ventricosus (family Araneidae, 0.3509), Dipoena castrata (0.2413, family Theridiidae), Argiope minuta (0.1836, family Araneidae), and Paracoelotes spinivulva (0.1760, family Amaurobiidae) have relatively strong activities among themselves. However, comparing to that of the brown recluse spider, Loxosceles recluse (1.814) in North America, the necrotizing shpingomyelinase activities of these Korean web‐building spiders are still very low. Based on our results, we may thus conclude that there would be little possibilities in South Korea to create serious medical problems caused by necrotizing arachnidism.     

Astacin family metallopeptidases and serine peptidase inhibitors in spider digestive fluid

Digestive fluid of the araneid spider Argiope aurantia is known to contain zinc metallopeptidases. Using anion-exchange chromatography, size-exclusion chromatography, sucrose density gradient centrifugation, and gel electrophoresis, we isolated two lower-molecular-mass peptidases, designated p16 and p18. The N-terminal amino acid sequences of p16 (37 residues) and p18 (20 residues) are 85% identical over the first 20 residues and are most similar to the N-terminal sequences of the fully active form of meprin (beta subunits) from several vertebrates (47-52% and 50-60% identical, respectively). Meprin is a peptidase in the astacin (M12A) subfamily of the astacin (M12) family. Additionally, a 66-residue internal sequence obtained from p16 aligns with the conserved astacin subfamily domain. Thus, at least some spider digestive peptidases appear related to astacin of decapod crustaceans. However, important differences between spider and crustacean metallopeptidases with regard to isoelectric point and their susceptibility to hemolymph-borne inhibitors are demonstrated. Anomalous behavior of the lower-molecular-mass Argiope peptidases during certain fractionation procedures indicates that these peptidases may take part in reversible associations with each other or with other proteins. A. aurantia digestive fluid also contains inhibitory activity effective against insect digestive peptidases. Here we present evidence for at least thirteen, heat-stable serine peptidase inhibitors ranging in molecular mass from about 15 to 32 kDa.     

Survey for potentially necrotizing spider venoms, with special emphasis on Cheiracanthium mildei

It has proven difficult to identify those spiders which cause necrotic lesions. In an effort to design a simple, inexpensive screening method for identifying spiders with necrotizing venoms, we have examined the venom gland homogenates of a variety of spider species for their ability to cause red blood cell lysis. Those venoms which were positive were further examined for the presence of sphingomyelinase D, and their ability to evoke necrotic lesions in the skin of rabbits. Sphingomyelinase D is known to be the causative agent of necrosis and red blood cell lysis in the venom of the brown recluse spider (Loxosceles reclusa), and our assumption was that this would be the same agent in other spider venoms as well. This did not prove to be the case. Of 45 species examined, only the venom of L. reclusa and Cheiracanthium mildei lysed sheep red blood cells. Unlike L. reclusa venom, however, C. mildei venom did not possess sphingomyelinase D nor did it cause necrotic lesions in the skin of rabbits. We present evidence suggesting that a phospholipase A2 is the hemolytic agent in C. mildei venom.     

Predation of wild spider monkeys at La Macarena,Colombia

The killing of an adult male spider monkey (Ateles belzebuth ) by a jaguar (Panthera onca) and a predation attempt by a puma (Felis concolor) on an adult female spider monkey have been observed at the CIEM (Centro de Investigaciones Ecológicas La Macarena), La Macarena, Colombia. These incidents occurred directly in front of an observer, even though it is said that predation under direct observation on any type of primate rarely occurs. On the basis of a review of the literature, and the observations reported here, we suggest that jaguars and pumas are likely to be the only significant potential predators on adult spider monkeys, probably because of their large body size.     

Screening of Necrotizing Spiders in Korea Using Nonradioactive Sphingomyelinase Assay (I) Wandering Spiders

There are now more than 40,000 identified spider species in the world, and considered about 100 species as actually dangerous to human. Spider bites cause a range of symptoms from simple swellings to disfiguring necrotic lesions, and occasionally death. While spider bites are not a major medical problem in Korea, it would be of great value to know which species of spiders pose a threat to human health. A middle molecular weight protein, sphingomyelinase D, has been identified in the venom of the brown recluse spider and strong evidence suggests that they have a major role in spider bite necrosis. For the identification of necrotizing species, we have investigated using recently developed non‐radioactive assay of sphingomyelinase for rapidly screening the necrotizing venoms. Here, we demonstrate the fetal toxicity of total 57 species (32 genera, 9 families) of the wandering spiders among 622 identified spider species in Korea. It has been revealed that two species of the Thomisidae spider, Ozyptila nongae (0.2467) and Diaea subdola (0.2020) have the strongest sphingomyelinase activities among themselves. In addition one species of the family Pisauridae, Dolomedes sulfureus (0.2341) has also relatively higher value comparing to other wandering spiders. However comparing to that of the brown recluse spider, Loxosceles reclusa (1.814) in North America the necrotizing activities of these Korean wandering species are still very low state, so there seems to be little possibilities to create serious medical problems by the necrotizing arachnidism in Korean peninsula.     

Feeding rate may affect dispersal in the orb-web spider Nephila clavata

Population parameters of the orb-web spider Nephila clavata were estimated at two sites with different spider densities (for 2 successive years at the high density site and 1 year at the low density site), and the relationship between survival rate and feeding conditions was examined. The rate of decrease of population density was almost constant over time and nearly the same at the two sites. Daily survival rate (sum of the effects of mortality and emigration) was low during August to early September and increased markedly thereafter. Daily immigration rate (number of immigrants/number of residents) was high during August to early September. Since there were strong negative correlations between survival and immigration rates, low survival rate seems to be caused by a high rate of emigration. Strong positive correlations were found between survival rate and feeding frequency (mean percent observed feeding). Analysis of covariance revealed that the parameters of the regression between survival rate and feeding frequency did not differ significantly among three occasions. These results suggest that feeding condition has a significant influence on dispersal activity of populations in this spider.     

崇明岛上沙地区蜘蛛种类及其动态分析

于2011年4月初到2013年10月底,在崇明岛上沙地区选择6个代表性区域,采用捕捉、扣管、扫网和陷阱法对蜘蛛种类进行系统调查和定点采集,共采集蜘蛛标本700头,初步鉴定出蜘蛛63种,隶属13科38属,另有8种鉴定到属。该地区蜘蛛优势种类是跳蛛科、肖蛸科、园蛛科、狼蛛科。文章介绍了蜘蛛种群年度消长规律,同时探讨了不同环境因子的变化对蜘蛛种群的影响,分析表明：蜘蛛种群密度与生境和农药的使用存在显著差异,蜘蛛适宜生存的温度在15℃～35℃,适易的相对湿度在65％-85％。     

Molecular characterization and evolutionary study of spider tubuliform (eggcase) silk protein

As a result of hundreds of millions of years of evolution, orb-web-weaving spiders have developed the use of seven different silks produced by different abdominal glands for various functions. Tubuliform silk (eggcase silk) is unique among these spider silks due to its high serine and very low glycine content. In addition, tubuliform silk is the only silk produced just during a short period of time, the reproductive season, in the spider's life. To understand the molecular characteristics of the proteins composing this silk, we constructed tubuliform-gland-specific cDNA libraries from three different spider families, Nephila clavipes, Argiope aurantia, and Araneus gemmoides. Sequencing of tubuliform silk cDNAs reveals the repetitive architecture of its coding sequence and novel amino acid motifs. The inferred protein, tubuliform spidroin 1 (TuSp1), contains highly homogenized repeats in all three spiders. Amino acid composition comparison of the predicted tubuliform silk protein sequence to tubuliform silk indicates that TuSp1 is the major component of tubuliform silk. Repeat unit alignment of TuSp1 among three spider species shows high sequence conservation among tubuliform silk protein orthologue groups. Sequence comparison among TuSp1 repetitive units within species suggests intragenic concerted evolution, presumably through gene conversion and unequal crossover events. Comparative analysis demonstrates that TuSp1 represents a new orthologue in the spider silk gene family.     

Structures and biological activities of three synaptic antagonists from orb weaver spider venom

The venom of Argiope aurantia, an orb weaver spider, contains a mixture of low molecular weight "argiotoxins", which block neuromuscular transmission in insects. Complete structure elucidation of three argiotoxins reveals common features; a hydrophilic, basic domain of arginine, a polyamine and asparagine is connected to an aromatic moiety contributed either by 4-hydroxyindole-3-acetic acid or 2,4-dihydroxyphenylacetic acid. Structural assignments of two argiotoxins are verified by chemical synthesis. The argiotoxins cause reversible paralysis when injected into insects and this is correlated with a stimulus-dependent inhibition of skeletal neuromuscular transmission at submicromolar concentrations.     

Spontaneous male death during copulation in an orb-weaving spider

Males of some cannibalistic species of spiders and insects appear to sacrifice themselves by allowing the female to eat them, and the adaptive significance of such drastic terminal reproductive investment has recently been demonstrated for a spider. Typically, the female has to kill the male, but it has been suggested that males of some species in the cannibalistic orb-weaving spider genus Argiope may die in copula without female 'collaboration'. Here, we provide the first experimental evidence to our knowledge of programmed sudden death after onset of copulation in males of the spider Argiope aurantia. Our observations reveal that males invariably die during the insertion of their second pedipalp, regardless of whether they mate with newly moulted, defenceless females or with older mature females that often attack them. We determined experimentally that the death of males is triggered immediately upon insertion of the second palp, when males become unresponsive, and heartbeat ceases within minutes of insertion. We discuss the possible adaptive significance of programmed death during copulation, and argue that male death has evolved in a context other than sexual cannibalism.     

Competing dwarf males: sexual selection in an orb-weaving spider

Hypotheses for the adaptive significance of extreme female-biased sexual size dimorphism (SSD) generally assume that in dimorphic species males rarely interfere with each other. Here we provide the first multivariate examination of sexual selection because of male-male competition over access to females in a species with 'dwarf' males, the orb-weaving spider Argiope aurantia. Male A. aurantia typically try to mate opportunistically during the female's final moult when she is defenceless. We show that, contrary to previous hypotheses, the local operational sex ratio (males per female on the web) is male-biased most of the season. Both interference and scramble competition occur during opportunistic mating, the former leading to significant selection for large male body size. Male condition and leg length had no effect on mating success independent of size. We discuss these findings in the context of the evolution of extreme female-biased SSD in this clade.     

'Partitagin' a hemorrhagic metalloprotease from Hippasa partita spider venom: role in tissue necrosis

The poisonous bite by Hippasa partita, a funnel web spider from the Indian subcontinent has been demonstrated to give rise to severe dermo- and myonecrosis. In this work a hemorrhagic metalloprotease, Partitagin was purified from H. partita venom by successive chromatography on Sephadex G-100, DEAE Sephadex A-50 and Biosep DEAE columns. SDS-PAGE, reversed phase HPLC on a C(4) column, N-terminal amino acid sequencing and MALDI-TOF mass spectrometry confirmed the homogeneity. Partitagin was assayed using fat free casein as substrate. EDTA, 1,10-phenanthroline and cyanide, inactivated it irreversibly while, EGTA, PMSF, leupeptin, pepstatin and aprotinin did not inhibit. The presence of Zn(+2) was confirmed by atomic absorption spectrometry. Partitagin caused hemorrhage when tested in a mouse model. Light microscopy of skin tissue sections at the site of injection revealed extensive damage of extracellular matrix (ECM) in which the basement membrane surrounding blood vessels and capillaries showing signs of extensive destruction and also loss of vessel wall integrity. Similar intense damage was also noticed in the ECM of muscle tissue sections but with no damage caused to myocytes. Partitagin showed specificity of action on the components of ECM and degraded collagen type-IV and fibronectin but not collagen type-I. Partitagin was devoid of edema, myotoxicity and lethality. This is the first report on the isolation and characterization of a toxin from spider venom in the Indian subcontinent.     

Homing by path integration in the spider Agelena labyrinthica Clerck

Moving about the web the spider Agelena labyrinthica continuously adjusts the prospective return angle. The amount of path integration was indicated by two compromise angles, return angle e and goal angle (Fig. 2). The spider was primed to one of two perpendicular light azimuths, L1 or L2. Subsequently, the discrete effects of a 90° change in light azimuth on the return direction were recorded (Fig. 3). When primed to L1, and the spider was exposed to L2: (1) while homebound, the deviation from straight home was clockwise and largest, (2) both while outbound and returning, the deviation was smallest (Fig. 4), (3) while outbound, either during the first or second half of the outbound run, the deviations were between those obtained in (1) and (2). When the spider was primed to L1, but given L2 while outbound and L1 again while homebound, Agelena deviated counterclockwise, the most with L2 on during the entire outbound run, and less when L2 was available only during a portion of the outbound run. The degree of adjustment of the home path direction is not correlated with the actual path length but with the shortest distance (bee line) between the two points during which the spider was exposed to one of the two light azimuths.On fellowship leave from Hunter College of the City University of New York, Department of Psychology, New York, NY 10021, USA     

蜘蛛毒素的研究进展及其相关应用

蜘蛛毒素含有多种化学成分,除毒性作用外这些物质对机体产生多重影响并具有多种活性。近年来,大量新的毒性组分不断被分离纯化,其结构和功能性作用被广泛深入研究,蜘蛛毒素成为了生物毒素领域新的研究热点。本文对蜘蛛毒素在离子通道作用机制方面的最新研究进展进行了阐述,同时还探讨了蜘蛛毒素在医学实践中新的应用和发展。     

Embryonic expression patterns of Wnt genes in the RTA-clade spider Cupiennius salei

Spiders represent widely used model organisms for chelicerate and even arthropod development and evolution. Wnt genes are important and evolutionary conserved factors that control and regulate numerous developmental processes. Recent studies comprehensively investigated the complement and expression of spider Wnt genes revealing conserved as well as diverged aspects of their expression and thus (likely) function among different groups of spiders representing Mygalomorphae (tarantulas), and both main groups of Araneae (true spiders) (Haplogynae/Synspermiata and Entelegynae). The allegedly most modern/derived group of entelegyne spiders is represented by the RTA-clade of which no comprehensive data on Wnt expression were available prior to this study. Here, we investigated the embryonic expression of all Wnt genes of the RTA-clade spider Cupiennius salei. We found that most of the Wnt expression patterns are conserved between Cupiennius and other spiders, especially more basally branching species. Surprisingly, most differences in Wnt gene expression are seen in the common model spider Parasteatoda tepidariorum (a non-RTA clade entelegyne species). These results show that data and conclusions drawn from research on one member of a group of animals (or any other organism) cannot necessarily be extrapolated to the group as a whole, and instead highlight the need for comprehensive taxon sampling.     

火炬树抗螨性生物测定

本文分别测定了二斑叶螨Tetranychus urticae Koch和山楂叶螨Tetranychus viennensis Zacher对火炬树的选择性,以及限制性条件下火炬树对叶螨的生存及产卵的影响。结果表明：在火炬树叶和苹果叶之间,二斑叶螨和山楂叶螨在2 h, 6 h, 10 h, 12 h对火炬树的选择率分别为9.5%, 6.5%,5%,5%和2%,5%,1%和0,显著低于对苹果的选择率。被限制于火炬树叶上生存的山楂叶螨144 h后死亡率100%,产卵力随生存时间增加而下降,至96 h后产卵终止; 被限制于火炬树叶上生存的二斑叶螨240 h后全部死亡,整个观察期内（24～216 h）始终没有产卵。据此认为： 叶螨强烈拒食火炬树,火炬树对叶螨具有生存、产卵抑制作用,但这种抑制作用呈现种间差异。     

Nonlinear responses of spider mites to drought-stressed host plants

Abstract. 1. This study examines the shape of the population response curve of the two-spotted spider mite, Tetranychus urticae Koch, feeding on bean plants, Phaseolus vulgaris L., which have been subjected to one of a range of drought stresses.
2. Abundance of spider mites declined at slight drought stress, increased at intermediate intensities of stress, and declined again at severe stress.
3. Fecundity showed the same pattern found for total abundance. Developmental rates of immatures, on the other hand, generally increased with stress intensity. Survivorship of immatures and adult females appeared unrelated to stress intensity.
4. These results indicate that responses of mites to drought-stressed beans are nonlinear. Such a pattern helps explain contradictory results obtained for spider mites and other arthropod herbivores.     

Ultrastructure of spider thread anchorages

Spiders attach silken threads to substrates by means of glue-coated nanofibers (piriform silk), spun into disc-like structures. The organization and ultrastructure of this nano-composite silk are largely unknown, despite their implications for the biomechanical function and material properties of thread anchorages. In this work, the ultrastructure of silken attachment discs was studied in representatives of four spider families with Transmission Electron Microscopy to facilitate a mechanistic understanding of piriform silk function across spiders. Based on previous findings from comparative studies of piriform silk gland morphology, we hypothesized that the fibre-glue proportion of piriform silk differs in different spiders, while the composition of fibre and glue fractions is consistent. Results confirmed large differences in the relative proportion of glue with low amounts in the orb weaver Nephila senegalensis (Araneidae) and the hunting spider Cupiennius salei (Ctenidae), larger amounts in the cobweb spider Parasteatoda tepidariorum (Theridiidae) and a complete reduction of the fibrous component in the haplogyne spider Pholcus phalangioides (Pholcidae). We rejected our hypothesis that glue ultrastructure is consistent. The glue is a colloid with polymeric and fluid fractions that strongly differ in proportions and assembly. We further confirmed that in all species studied both dragline and piriform silk fibres do not make contact with the environmental substrate. Instead, adhesion is established by a thin dense skin layer of the piriform glue. These results advance our understanding of piriform silk function and the interspecific variation of its properties, which is significant for spider biology, web function and the bioengineering of silk.