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.     

Anti-predator defences of Argiope appensa (Araneae, Araneidae), a tropical orb-weaving spider

Anti-predator defence behaviour of Argiope appensa (Fuesslin) (Araneae, Araneidae) was studied in the laboratory. The most frequent response of adults and large juveniles of A. appensa to disturbance was pumping, a behaviour during which this web-building spider moved its body rapidly up and down with its legs remaining on the silk. When disturbed, small juveniles differed from adult females and larger juveniles by often dropping from the web instead of pumping. Argiope appensa sometimes put its web betweeen itself and stimuli from potential predators by shuttling from one side of the hub to the other. Argiope appensa occasionally tugged on the web but this behaviour appeared to be primarily a component of prey-catching sequences instead of defence. Experiments were carried out to determine the types of stimuli that elicited pumping. Lightly touching the spider or its web, forcefully hitting the web, and air movement elicited pumping but there was no evidence that chemical stimuli from potential predators were important.     

Spidroins from the Brazilian spider Nephilengys cruentata (Araneae: Nephilidae)

Spiders produce up to six different kinds of silk, each one for a specific biological function. Spider silks are also known for their unique mechanical properties. The possibility of producing new materials with similar properties motivated research on these silk proteins (spidroins). Using expression sequence tags, we identified four spidroins produced by major ampullate, minor ampullate, flagelliform and tubuliform silk glands from the Brazilian spider Nephilengys cruentata (Araneae: Nephilidae). The new protein sequences showed substantial similarity to other spidroins previously described, with high content of alanine and glycine due to the presence of the highly repetitive motifs (polyAla, (GA)_n, (GGX)_n, (GPGGX)_n). Similarities among sequences were also observed between the different spidroins with the exception of tubuliform spidroin, which presents a unique complex amino acid sequence with high amounts of serine and low amounts of glycine.     

Activities against hemostatic proteins and adrenal gland ultrastructural changes caused by the brown widow spider Latrodectus geometricus (Araneae: Theridiidae) venom

Brown widow spider (BrWS) (Latrodectus geometricus) venom produces intense systemic reactions such as cramps, harsh muscle nociceptive, nauseas, vomiting and hypertension. The proposed pathogenic mechanisms resulting in these accidents have principally been damages occurring at the nervous system. However, it is suspected that there is also damage of the adrenal glands, as a result of the experimental animal's clinical manifestations, which developed symptoms compatible with acute adrenal insufficiency. We have currently found that the adrenal gland is damaged by this venom gland homogenates (VGH) producing severe alterations on cortex cells resulting in death by acute adrenal insufficiency. In general, the ultrastructural study on the glands of mice under transmission electronic microscopy observations showed alterations in the majority of the intracellular membranes within 3 to 24 h. BrWSVGH also showed specific actions on extracellular matrix proteins such as fibronectin, laminin and fibrinogen. In addition, zymogram experiments using gelatin as substrates detected gelatinolytic activity. The molecular exclusion fractionation of crude BrWSVGH resulted in 15 fractions, of which F1 and F2 presented α/β-fibrinogenase and fibronectinolytic activities. Fractions F6, F14 and F15 showed only α-fibrinogenase activity; in contrast, the gelatinolytic action was only observed in fraction F11. Only metalloproteinase inhibitors abolished all these proteolytic activities. Our results suggest that adrenal cortex lesions may be relevant in the etiopathogenesis of severe brown widow spider envenoming. To our knowledge, this is the first report on adrenal gland damages, fibrinogenolytic activity and interrelations with cell-matrix adhesion proteins caused by L.geometricus VGH. The venom of this spider could be inducing hemostatic system damages on envenomed patients.     

A phylogenetic analysis of the orb-weaving spider family Araneidae (Arachnida, Araneae)

We present the first cladistic analysis focused at the tribal and subfamily level of the orb-weaving spider family Araneidae. The data matrix of 82 characters scored for 57 arancid genera of 6 subfamilies and 19 tribes (and 13 genera from 8 outgroup families) resulted in 16 slightly different, most parsimonious trees. Successive weighting corroborated 62 of the 66 informative nodes on these cladograms; one is recommended as the 'working' araneid phylogcny. The sister group of Araneidae is all other Araneoidea. Araneidae comprises two major clades: the subfamily Araneinae, and the 'argiopoid' clade, which includes all other subfamilies and most tribes (((Gasteracanthinae, Caerostreae), (((Micratheninae, Xylcthreae), Eruyosaccus ), (Eurycorminae, Arciinae)), Cyrlarachninae), ((Argiopinae, Cyrtophorinae), Arachnureae)). Cyrtarachneae and Mastophoreae are united in a new subfamily, Cyr-tarachninae. The spiny orb-weavers alone (Gasteracanthinae and Micratheninae) are not monophyletic. The mimetid subfamily Arciinae and the 'tetragnathid' genus Zygiella are araneids, but .Nephila (and other tetragnathids) are not. On the preferred tree, web decorations (stabilimenta) evolved 9 times within 15 genera, and were lost once. The use of silk to subdue prey evolved once in cribellate and four times in ecribillate orb weavers. Sexual size dimorphism evolved once in nephilines, twice in araneids, and reverted to monomorphism five times. Evolution in other genitalic and somatic characters is also assessed; behavioral and spinneret features arc most consistent (male genitalia, leg and prosomal features least consistent) on the phylogeny.     

Bacterial production of latarcin 2a, a potent antimicrobial peptide from spider venom

Natural venoms are promising sources of candidate therapeutics including antibiotics. A recently described potent antimicrobial peptide latarcin 2a (Ltc 2a) from Lachesana tarabaevi spider venom shows a broad-spectrum antibacterial activity. This peptide consists of 26 amino acid residues and therefore its production using chemical synthesis, although trivial, is costly. We describe an easy approach to Ltc 2a production in Escherichia coli using the conventional fusion partner thioredoxin. Latarcin 2a synthetic gene was cloned into the expression vector pET-32b, which was then used to transform E. coli BL21(DE3) strain. His-tagged fusion purification was achieved using metal-chelate affinity chromatography. Since no methionine residues are present in the latarcin 2a sequence, cyanogen bromide could be effectively utilized to separate the target product from the carrier protein. Reverse-phase HPLC was used as the final step of purification; the final yield was 3 mg/L of bacterial culture. To increase the yields, we attempted incorporation of Ltc 2a tandem repeats into the fusion protein; however, production rates greatly decreased due to enhanced fusion toxicity. Moreover, we probed constructs to produce an Ltc 2a dimer and the Ltc 2a propeptide to study their functional properties. Recombinant peptides were produced at appreciable yields and biological tests to determine their activities were performed. Latarcin 2a is the first linear peptide from spider venom and one of the first membrane-active peptides from venomous animals to be biosynthetically produced.     

'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.     

Trophic niche,capture efficiency and venom profiles of six sympatric ant‐eating spider species (Araneae: Zodariidae)

The arms race between specialist predators and their prey has resulted in the evolution of a variety of specific adaptations. In venomous predators, this can include venom composition, particularly if predators are specialized on dangerous prey. Here, we performed an integrative study using six species of highly specialized ant‐eating spiders of the genus Zodarion to investigate their phylogeny, realized trophic niche, efficacy in the capture of various ant species and venom composition. Data on natural diet obtained by next‐generation sequencing and field observations showed that the six Zodarion species exploit different ant species. Their phylogeny, based on mitochondrial and nuclear genes, correlated with the composition of their natural prey, indicating that closely related Zodarion species specialize on similar ant species. Prey‐capture parameters differed among Zodarion species suggesting prey‐specific efficacy. Similarly, the venom profiles of both low and high molecular compounds differed among species. Only the profiles of low molecular compounds were correlated with capture efficacy parameters, suggesting that the venom of Zodarion spiders contains prey‐specific components. Our study suggests that Iberian Zodarion spiders are specialized on particular ant species.     

Hainantoxin-Ⅱ的分离纯化及其结构与功能分析(英文)

从分布于我国海南省的海南捕鸟蛛(Haplopelma hainanum)毒素中,利用阳离子交换色谱和反相高效液相色谱分离得到一种多肽神经毒素,命名为Hainantoxin-Ⅱ(HnTx-Ⅱ)。MALDI-TOF质谱分析表明该多肽毒素相对分子质量为4253.135,其氨基酸序列经Edman降解测序为LFECS VSCEI EKEGN KDCKK KKCKG GWKCK FNMCV KV,其中的6个Cys形成3对二硫键。同源性搜索表明,HnTx-Ⅱ与Huwentoxin-Ⅱ(HwTx-Ⅱ)的同源性高达91%,仅有3个氨基酸残基不同。HwTx-Ⅱ为从虎纹捕鸟蛛中提取的杀虫肽,该多肽具有独特的结构模体。活性分析表明HnTx-Ⅱ与HwTx-Ⅱ具有相似的生物学活性。经腹腔注射HnTx-Ⅱ,美洲蜚蠊可被麻痹,其ED50为16μg/g;而当剂量增加到60μg/g时,可立即杀死美洲蜚蠊,其杀死昆虫活性明显强于HwTx-Ⅱ。此外,HnTx-Ⅱ经脑室注射可杀死小鼠,其LD50为1.41μg/g,该活性却明显低于HwTx-Ⅱ。这两种多肽毒素的结构与功能的差异为进一步阐明多肽毒素的结构与功能之间的关系提供良好的研究模型。     

Function of bright coloration in the wasp spider Argiope bruennichi (Araneae: Araneidae)

There are two major competing explanations for the counter-intuitive presence of bright coloration in certain orb-web spiders. Bright coloration could lure insect prey to the web vicinity, increasing the spider's foraging success. Alternatively, the markings could function as disruptive camouflage, making it difficult for the insect prey to distinguish spiders from background colour variation. We measured the prey capture rates of wasp spiders, Argiope bruennichi, that were blacked out, shielded from view using a leaf fragment, or left naturally coloured. Naturally coloured spiders caught over twice the number of prey as did either blacked-out or leaf-shielded spiders, and almost three times as many orthopteran prey. Spectrophotometer measurements suggest that the bright yellow bands on the spider's abdomen are visible to insect prey, but not the banding on the legs, which could disguise the spider's outline. Thus, our results provide strong support for the hypothesis that bright coloration in the wasp spider acts as a visual lure for insect prey and weak support for the hypothesis that the arrangement of the banding pattern across the spider's body disguises the presence of the spider on the web.     

Spiderling sex ratio and maternal investment in the bolas spider Mastophora cornigera (Araneae,Araneidae)

Male Mastophora cornigera exit egg sacs as adults, which allowed us to determine spiderling sex ratios and patterns of maternal investment in this species. We collected 15 egg sacs produced by seven mothers, which yielded 1945 emergent spiderlings which were sexed, 1850 of which were weighed. Two emergent broods were significantly male and female biased and were unaffected by pre-emergence mortality. The weights of male and female spiderlings differed in eight broods, with males and females being heavier in four cases each. Five of these broods were derived from multiple egg sac sets produced by one mother, and in each case, the total mean male and female spiderling weights for all broods in a set were biased in the same direction as the biased brood(s) within that set. Mean emergent spiderling weight was independent of brood size and sex ratio for both males and females. Despite such independence, sex allocation in M. cornigera can favor sons, daughters, or both equally, and by numbers, by weight, or both at once. The proximate mechanisms and adaptive significance of such variability is unknown. We also review evidence for gender-biased allocations in arachnid offspring and suggested mechanisms for their applicability to M. cornigera.     

Heterozygosity and fitness in a California population of the labyrinth spider Metepeira ventura (Araneae, Araneidae)

Abstract. The relationship between individual heterozygosity and characteristics likely to be associated with fitness was investigated in the labyrinth spider Metepeira ventura . Adult females and their egg sacs were collected at a coastal site in southern California, and three measures of bodily condition (carapace width, weight, residual index) and six measures of reproductive output (number of egg sacs, variation in egg number among sacs [coefficient of variation], total number of eggs, mean eggs/sac, mean eggs/sac divided by carapace width, mean eggs/sac divided by weight) were determined for each spider. The sample was polymorphic at three allozyme loci that were in Hardy–Weinberg equilibrium, and individual females were heterozygous at up to two of the three loci, forming three heterozygosity classes (0, 1, and 2). None of the bodily condition measures were significantly related to the number of heterozygous loci, while four of the reproductive output estimators (total number of eggs, mean eggs/sac, mean eggs/sac divided by carapace width, mean eggs/sac divided by weight) were significantly influenced by heterozygosity. In each significant case, values for class 2 females were less than those for class 0 and 1 females, whose values were usually more similar. Thus, while female bodily condition was comparable among classes, the most heterozygous females produced fewer total eggs and eggs per sac than their less heterozygous peers. The fact that females of M. ventura engage in a reproductive investment-number trade-off suggests that high-variability and low-variability females may be pursuing distinct reproductive strategies in the wild, with more heterozygous females being K -selected (smaller clutches, heavier eggs) and more homozygous females being r -selected (larger clutches, lighter eggs). Further investigation will be needed to assess more fully the fitness value of heterozygosity in M. ventura .     

A new combination expands the range of the African araneid spider Singafrotypa (Araneae, Araneidae)

Study of the syntype of Larinioides subinermis, a species known from Ethiopia only, revealed that it actually belongs to Singafrotypa Benoit, 1962. We redescribe Singafrotypa subinermis (Caporiacco, 1940), comb. n., and provide a key to females of four species belonging to Singafrotypa. A distribution map for all species is provided.     

Physiological costs during the first maternal care in the wolf spider Pardosa saltans (Araneae,Lycosidae)

Many arachnids like other terrestrial arthropods, provide extensive maternal care. Few studies have quantified the underlying physiological costs of maternal care. We investigated how maternal care affects the free-moving wolf spider’s (Pardosa saltans) energy requirements. We described in detail their basic reproduction biology (i.e. carrying cocoon and young) and we evaluated the variation in the females’ energy reserves during maternal care. Our results show that mothers guard eggs until hatching and then guard their spiderlings for 27–30 more days. Laboratory observations indicated that spiderlings start leaving the maternal abdomen gradually 5–7 days after hatching. Females carry an egg sac (cocoon) that can weigh up to 77% of their post-reproduction weight and carry young that weigh 87–100% of their body mass. Females lost weight over time despite regular food intake, while carrying cocoon and young; but their weights increased gradually during the dispersal of young. The contributions of proteins, glucose and triglycerides to maintain females’ energy were calculated. Their energetic state varied during maternal care, in particular lipid levels declined, during the care of spiderlings when the females’ predatory behaviour was inhibited. Our results show that the maternal care provided by P. saltans females is particularly costly physiologically, during the 30 days following egg sac formation and development of spiderlings, even when food is available.     

Tetrablemmidae (Arachnida, Araneae), a spider family newly recorded from China

The family Tetrablemmidae is reported for the first time from China, with five new species and one new genus. Lehtinenia gen. n., which is erected to accommodate Lehtinenia bicornis sp. n., is characterized by the modified embolus, special modifications on chelicerae, and a Tetrablemma-type vulva. The other four new species are: Ablemma prominens sp. n., Brignoliella caligiformis sp. n., Brignoliella maoganensis sp. n., and Tetrablemma brevidens sp. n., all collected from caves. A phylogenetic analysis of the subfamily Tetrablemminae based on 41 morphological characters shows that the tribe Brignoliellini is the most basal group in the subfamily, rather than the sister group to the tribe Fallablemmini. Lehtinenia gen. n. and the genera Ablemma, Sulaimania, and Maijana together form a monophyletic group.     

Organization of the spinnerets and spigots in the orb web spider,Argiope bruennichi (Araneae: Araneidae)

Although the basic taxonomic characteristics usually remain unchanged, some spinning apparatuses undergo consistent adaptive variations. As the presence of additional protuberances known as nubbins and tartipores have caused disagreements regarding some Araneidae spiders, more detailed definitions on the cuticular structures have recently been proposed. Reflecting this definition, microstructural organization of silk spinning apparatuses in the orb web spider Argiope bruennichi were reconsidered using field emission scanning electron microscopy. Among the seven kinds of functional spigots in females, it was revealed that two types (major ampullates and pyrifoms) are located on anterior spinnerets and another five types are distributed on median (minor ampullates, tubuliforms and aciniforms) or posterior (tubuliforms, flagelliforms, aggregates and aciniforms) spinnerets, respectively. In addition to functional spigots, cuticular remnants of the nubbins and the tartipores were found on the spinning fields, but the number of tartipores on each spinneret varied among individuals based on maturity. Nevertheless, three kinds of cuticular protuberances of ampullate silk glands were clearly visible at both the anterior and median spinnerets.     

The ultrastructure of the book lungs of the Italian trap-door spider Cteniza sp. (Araneae,Mygalomorphae, Ctenizidae)

The fine structure of book lungs is not homogeneous across Arachnids and is considered phylogenetically informative, however few reports on the ultrastructural features of this organ have been published. In this study, we examined the general morphology and ultrastructure of adult spiders of the genus Cteniza. The respiratory system of Cteniza sp. consists of two pairs of well-developed book lungs, which is considered indicative of primitive spiders. The general organization of the book lungs is similar to that described for other arachnids and consists of leaves of alternating air and hemolymph channels. The air channels are lined with cuticle and open to an atrium that leads to a slit-like spiracle. The air channels are held open by cuticular trabeculae. The space holders in the hemolymph channels are pillar trabeculae formed by two cells from the opposed walls. The pillar cells have a complex ultrastructure that includes an interdigitating connection, gap junctions, microtubules and hemidesmosomes. These features apparently help strengthen the pillar cells and their interconnections with each other and the underlying cuticle. The cytoskeleton resembles that of arthropod tendon cells where substantial structural support is needed.     

Early development of the anterior body region of the grey widow spider Latrodectus geometricus Koch, 1841 (Theridiidae, Araneae)

We document the early morphogenesis of Latrodectus geometricus, particularly of the anterior body region. Significant changes in the development of the external prosomal structures revealed with scanning electron microscopy (SEM) images include: (1) reorganisation of each pre-cheliceral lobe by subdivision and internalisation of its central area; (2) shortening of the ventro-median bridge connecting the pre-cheliceral lobes and its eventual disappearance; (3) appearance and expansion of a prospective mouth region between the pre-cheliceral lobes with a recessed median area surrounded by lip-like borders, the anterior lip-part developing into the hypostome; (4) reduction of the mouth region to an area around the hypostome and the lip-like latero-posterior border of the mouth opening; (5) change of the position of the mouth region from anterior to the insertions of the chelicerae to posterior to them; (6) eventual shortening of the mouth opening to a slit overhung by the hypostome; (7) origination of the prosomal shield from the anterior margin of the pre-cheliceral lobes and the tergal portions of the four posterior-most prosomal segments; and (8) expansion of a ‘ventral sulcus’ from the cheliceral to the fifth opisthosomal segment separating the sides of these segments. Embryonic features are compared across the Chelicerata and discussed briefly in a phylogenetic context.     

Cannibalism and kin recognition in Delena cancerides (Araneae: Sparassidae), a social huntsman spider

Social behaviour in spiders is rare: of the 39 000 species of spiders known, only 23 are considered to be cooperatively social. Delena cancerides is a social species of the huntsman spider that is endemic to Australia. This species is virtually unique among social spiders, having evolved social behaviour in the absence of a snare web. It is thought that this form of social behaviour in D. cancerides has evolved via the sub-social route, that is, the extension of an ancestrally occurring period of maternal care and the delayed dispersal of juveniles. Most social spiders show no aggression towards non-kin conspecifics, prompting suggestions that spiders cannot recognize kin; however, D. cancerides individuals are highly aggressive towards conspecifics introduced from outside their own colony. In order to determine whether selective aggression in D. cancerides has its basis in kin recognition, tolerance behaviour was assessed in the context of kinship and size. We observed that, in general, juveniles preferred to starve than engage in cannibalism of any conspecifics, related or not. However, where cannibalism did occur, non-kin were preferentially eaten, indicating that this species is clearly capable of kin recognition. Size thresholds were also established, below which juveniles are tolerated by adults and above which aggressive interactions leading to death occur. We conclude that kin recognition and juvenile dispersal explain the uncharacteristically high levels of genetic polymorphism in this species.