Bacteria inhabiting spider webs enhance host silk extensibility.

Spider silk is a promising material with great potential in biomedical applications due to its incredible mechanical properties and resistance to degradation of commercially available bacterial strains. However, little is known about the bacterial communities that may inhabit spider webs and how these microorganisms interact with spider silk. In this study, we exposed two exopolysaccharide-secreting bacteria, isolated from webs of an orb spider, to major ampullate (MA) silk from host spiders. The naturally occurring lipid and glycoprotein surface layers of MA silk were experimentally removed to further probe the interaction between bacteria and silk. Extensibility of major ampullate silk produced by Triconephila clavata that was exposed to either Microbacterium sp. or Novosphigobium sp. was significantly higher than that of silk that was not exposed to bacteria (differed by 58.7%). This strain-enhancing effect was not observed when the lipid and glycoprotein surface layers of MA silks were removed. The presence of exopolysaccharides was detected through NMR from MA silks exposed to these two bacteria but not from those without exposure. Here we report for the first time that exopolysaccharide-secreting bacteria inhabiting spider webs can enhance extensibility of host MA silks and silk surface layers play a vital role in mediating such effects.

Testing sea urchin and green sea turtle consumption of the allelopathic macroalga Galaxaura divaricata.

Galaxaura divaricata is a partially calcified macroalga that hampers coral recruitment, growth, and recovery via the excretion of allelopathic secondary metabolites. Herbivorous fishes are not major consumers of Galaxaura spp. and there is a need to understand feeding preferences for Galaxaura divaricata in other macroherbivores, like sea urchins and green sea turtles that could act as potential controlling agents. Under certain environmental conditions, G. divaricata can proliferate and overgrow degraded reefs for several years, as documented for several coral patch reefs in the lagoon of Dongsha Atoll, South China Sea. This study aimed to experimentally test the feeding preferences of five species of sea urchin and two individual green sea turtles, Chelonia mydas, for G. divaricata. Specifically, we quantified and compared the consumption rates of the allelopathic G. divaricata with Gracilaria edulis, a nonallelopathic, fleshy red alga, known to be highly favored by herbivores. Results showed that the five urchin species fed on both G. edulis and G. divaricata. However, urchins consumed 2-8 times less wet weight of G. divaricata (range 0.3-3.1 g urchin-1 24 h-1) compared to G. edulis (range 0.6-18 g urchin-1 24 h-1), suggesting that urchin grazing may exert some control on G. divaricata abundance but is likely ineffective for a large-scale removal of the alga. Further, both green sea turtles avoided G. divaricata and selectively fed on G. edulis. More experiments are needed to test the potential role of herbivores in controlling the overgrowth of coral competitive and allelopathic macroalgae, like Galaxaura on coral reefs.

Loss of sea turtle eggs drives the collapse of an insular reptile community.

Marine subsidies are vital for terrestrial ecosystems, especially low-productivity islands. However, the impact of losing these subsidies on the terrestrial food web can be difficult to predict. We analyzed 23 years of survey data from Orchid Island to assess the consequences of the abrupt loss of an important marine subsidy. After climate-driven beach erosion and predator exclusion efforts resulted in the abrupt loss of sea turtle eggs from the terrestrial food web, predatory snakes altered their foraging habitats. This increased predation on other reptile species in inland areas, resulting in population declines in most terrestrial reptile species. Comparisons with sea turtle-free locations where lizard populations remained stable supported these findings. Our study emphasizes the cascading effects of generalist predators and the unintended consequences of single-species conservation, highlighting the importance of understanding species interconnectedness and considering potential ripple effects in marine-dependent insular ecosystems.

Novel field observations of coral reef fishes feeding on epiphytic and epizoic organisms associated with the allelopathic seaweed Galaxaura divaricata.

In degraded coral reef ecosystems, allelopathic macroalgae have received increasing attention from marine ecologists because their secondary metabolites (also known as allelochemicals) kill corals that grow adjacent to them and weaken the recovery of degraded reefs. One well-known coral-killing macroalga is the calcareous red seaweed Galaxaura. However, our knowledge of how coral reef fishes interact with allelopathic algae like Galaxaura is very limited. Here, we documented novel observations of feeding interactions of 17 species of coral reef fishes (herbivorous and carnivorous) with the filamentous Galaxaura divaricata on degraded lagoon patch reefs in Dongsha Atoll (South China Sea). Video analyses showed that territorial farming damselfishes (i.e., Dischistodus perspicillatus, D. prosopotaenia, Hemiglyphidodon plagiometopon, Pomacentrus grammorhynchus, P. adelus, and Neoglyphidodon nigroris) and juvenile parrotfishes (Scarus schlegeli, S. ghobban, S. rivulatus, and Chlorurus spilurus) likely used G. divaricata as a feeding substratum. Further, microscopic analyses revealed that the filamentous surface of G. divaricata harbored a wealth of epiphytic microalgae, such as filamentous cyanobacteria (i.e., Leptolyngbya, Lyngbya, Rivularia, Oscillatoria, and Stigonema), diatoms (i.e., Synedra, Nitzschia, Mastogloia, and Pleurosigma), and filamentous red algae (i.e., Heterosiphonia), suggesting that these fishes targeted the nutrient-rich microscopic epiphytes rather than the nutrient-poor host. Juvenile benthic carnivores (i.e., Labridae, Parupeneus multifasciatus, and Meiacanthus grammistes) form feeding assemblages with roving parrotfishes to feed on small invertebrates (i.e., amphipods, copepods, isopods, gastropods, and polychaetes) associated with G. divaricata. Given that coral reef fishes appear to target the epiphytes associated with Galaxaura rather than the alga itself, these observations thus substantiate the threat posed by the overgrowth of G. divaricata to coral recovery in degraded reef systems due to the lack of natural grazers.

Geographic distance, sedimentation, and substrate shape cryptic crustose coralline algal assemblages in the world's largest subtropical intertidal algal reef.

Algal reefs, concreted by crustose coralline algae (CCA), are the main biotic reefs in temperate waters but rare in the subtropics and tropics. The world's largest known intertidal algal reef in the subtropics is the Taoyuan Algal Reef (TAR) located in the northwestern coast of Taiwan. The biodiversity and ecology of the TAR are scarcely explored, and now the reef is imperiled by industrialization. Here, we document cryptic species of CCA in Taiwan, particularly the TAR, by sequencing the psbA genes of over 1800 specimens collected across Taiwan. We also examine the ecological background of the TAR by surveying its benthic composition and measuring its environmental parameters. Our data reveal that the TAR harbours a high diversity of cryptic CCA species (27 molecular operational taxonomic units, or mOTUs), many of which are potentially new to science (18 mOTUs) and/or endemic to the TAR (9 mOTUs). Comparing the CCA species inventory of the TAR with the rest of Taiwan shows that the TAR represents a unique hotspot of CCA taxa in the waters of Taiwan. Our analyses show that variation in the CCA assemblages in the TAR is associated with geographic distance, sedimentation, and substrate type (for example, reef vs. hermit crab shell), suggesting that dispersal limitation and contemporary environmental selection shape the CCA assemblages in the TAR. The data from this study can inform the monitoring of human impacts on the health of the TAR and contribute to our understanding of the ecological processes underlying algal reef development.

Short-term exposure of anticoagulant rodenticides leads to the toxin accumulation from prey (Rattus losea) to predator (Elanus caeruleus).

Rodenticides are widely used around the world since the 1950s. In Taiwan, an anti-rodent operation initiated 1977 and became a regular action annually implied by the government until 2014. This anti-rodent operation caused many animals of non-target species being exposed by rodenticides and became an environmental issue. The Black-winged Kite (Elanus caeruleus) is a small-sized diurnal raptor widely distributed in the Old World continent. Since 2000, a newly colonized population of this species occurred in Taiwan. Although the Black-winged Kites may suffer from the threats of rodenticides, the population is still growing and soon became the most abundant raptor in farmlands of Taiwan. Whether the Black-winged Kite accumulates higher anticoagulant rodenticide residues than other raptors are still unclear. In this study, liver samples of Black-winged Kites were collected from 2013 to 2016, when the detected residues of anticoagulant rodenticides increased annually. The concentration of residue rodenticide was above 0.2 ppm among 30% of the detected samples, which is the toxicity threshold concentration of other raptors. In the meanwhile, the lesser ricefield rat (Rattus losea), the most common prey of Black-winged Kites, also extended the survival period after fed on rodenticide. The longer survival days after being poisoned can enhance the predation opportunity of raptors, thus affect the accumulated rodenticides in the raptors. This study demonstrates that the Black-winged Kite has higher concentration of anticoagulant rodenticide than most other raptors, which provide the case that the raptor can quickly accumulate rodenticide residues within a short period of time.

Size-related increase in inducible mechanical variability of major ampullate silk in a huntsman spider (Araneae: Sparassidae).

Most spiders use major ampullate silk (MAS) to perform many functions across their lifetimes, including prey capture, vibratory signal detection, and safety/dragline. To accommodate their various needs, adult spiders can use inducible variability to tailor MAS with specific mechanical properties. However, it is currently unknown whether this inducible mechanical variability develops gradually or remains consistent across spider size. Supercontraction -a process by which "native-state" silk fibers axially shrink when exposed to water or high humidity-can be used to reveal the extent of inducible variability in MAS. Supercontraction removes some processing effects that occur during the spinning of the solid fiber from its liquid precursor by allowing a native-state MAS fiber to return to a low energy "ground-state". Here, we examined the relative extent of inducible variability of MAS across spider size by assessing supercontraction properties and the difference between ground- and native-state MAS tensile properties using silk from the huntsman spider Heteropoda venatoria (Sparassidae). Stiffness of forcibly pulled native-state silk increased by 200% with spider size. After exposure to 90% RH and subsequent supercontraction, axial shrinkage of native-state silk fibers increased by 15% in larger spiders. Supercontracted, ground-state fibers demonstrated a 200% increase in extensibility across spider size. Our results indicate a gradual increase in inducible variability of MAS mechanical properties across spider size potentially caused by shifts in internal processing or chemical composition. These findings imply both development of inducible variability and changes in use of MAS as a safety line or aiding jumps across a spider's lifetime.

Sum of fears among intraguild predators drives the survival of green sea turtle (Chelonia mydas) eggs.

Ecologists have long theorized that apex predators stabilize trophic systems by exerting a net protective effect on the basal resource of a food web. Although experimental and observational studies have borne this out, it is not always clear what behavioural mechanisms among the trophically connected species are responsible for this stability. Fear of intraguild predation is commonly identified as one such mechanism in models and mesocosm studies, but empirical evidence in natural systems remains limited, as the complexity of many trophic systems renders detailed behavioural studies of species interactions challenging. Here, we combine long-term field observations of a trophic system in nature with experimental behavioural studies of how all the species in this system interact, in both pairs and groups. The results demonstrate how an abundant, sessile and palatable prey item (sea turtle eggs, Chelonia mydas) survives when faced by three potential predators that all readily eat eggs: an apex predator (the stink ratsnake, Elaphe carinata) and two mesopredators (the brown rat, Rattus norvegicus, and kukri snake, Oligodon formosanus). Our results detail how fear of intraguild predation, conspecific cannibalism, habitat structure and territorial behaviour among these species interact in a complex fashion that results in high egg survival.

Trans-marine dispersal inferred from the saltwater tolerance of lizards from Taiwan.

Dehydration and hypersalinity challenge non-marine organisms crossing the ocean. The rate of water loss and saltwater tolerance thus determine the ability to disperse over sea and further influence species distribution. Surprisingly, this association between physiology and ecology is rarely investigated in terrestrial vertebrates. Here we conducted immersion experiments to individuals and eggs of six lizard species differently distributed across Taiwan and the adjacent islands to understand if the physiological responses reflect the geographical distribution. We found that Plestiodon elegans had the highest rate of water loss and the lowest saltwater tolerance, whereas Eutropis longicaudata and E. multifasciata showed the lowest rate of water loss and the highest saltwater tolerance. Diploderma swinhonis, Hemidactylus frenatus, and Anolis sagrei had medium measurements. For the eggs, only the rigid-shelled eggs of H. frenatus were incubated successfully after treatments. While, the parchment-shelled eggs of E. longicaudata and D. swinhonis lost or gained water dramatically in the immersions without any successful incubation. Combined with the historical geology of the islands and the origin areas of each species, the inferences of the results largely explain the current distribution of these lizards across Taiwan and the adjacent islands, pioneerly showing the association between physiological capability and species distribution.

Adhesion of spider cribellate silk enhanced in high humidity by mechanical plasticization of the underlying fiber.

The disruptive nature of water presents a significant challenge when designing synthetic adhesives that maintain functionality in wet conditions. However, many animal adhesives can withstand high humidity or underwater conditions, and some are even enhanced by them. An understudied mechanism in such systems is the influence of material plasticization by water to induce adhesive work through deformation. Cribellate silk is a dry adhesive used by particular spiders to capture moving prey. It presents as a candidate for testing the water plasticization model as it can remain functional at high humidity despite lacking an aqueous component. We performed herein tensile and adhesion tests on cribellate threads from the spider, Hickmania troglodytes; a spider that lives within wet cave environments. We found that the work of adhesion of its cribellate threads increased as the axial fibre deformed during pull-off experiments. This effect was enhanced when the silk was wetted and as spider body size increased. Dry threads on the other hand were stiff with low adhesion. We rationalized our experiments by a series of scaling law models. We concluded that these cribellate threads operate best when the nanofibrils and axial fibers both contribute to adhesion. Design of future synthetic materials could draw inspiration from how water facilitates, rather than diminishes, cribellate silk adhesion.

Mechanical and structural properties of major ampullate silk from spiders fed carbon nanomaterials.

The dragline silk of spiders is of particular interest to science due to its unique properties that make it an exceptional biomaterial that has both high tensile strength and elasticity. To improve these natural fibers, researchers have begun to try infusing metals and carbon nanomaterials to improve mechanical properties of spider silk. The objective of this study was to incorporate carbon nanomaterials into the silk of an orb-weaving spider, Nephila pilipes, by feeding them solutions containing graphene and carbon nanotubes. Spiders were collected from the field and in the lab were fed solutions by pipette containing either graphene sheets or nanotubes. Major ampullate silk was collected and a tensile tester was used to determine mechanical properties for pre- and post-treatment samples. Raman spectroscopy was then used to test for the presence of nanomaterials in silk samples. There was no apparent incorporation of carbon nanomaterials in the silk fibers that could be detected with Raman spectroscopy and there were no significant improvements in mechanical properties. This study represents an example for the importance of attempting to replicate previously published research. Researchers should be encouraged to continue to do these types of investigations in order to build a strong consensus and solid foundation for how to go forward with these new methods for creating novel biomaterials.

Uncoiling springs promote mechanical functionality of spider cribellate silk.

Composites, both natural and synthetic, achieve novel functionality by combining two or more constituent materials. For example, the earliest adhesive silk in spider webs - cribellate silk - is composed of stiff axial fibers and coiled fibers surrounded by hundreds of sticky cribellate nanofibrils. Yet, little is known of how fiber types interact to enable capture of insect prey with cribellate silk. To understand the roles of each constituent fiber during prey capture, we compared the tensile performance of native-state and manipulated threads produced by the cribellate spider Psechrus clavis, and the adhesion of native threads along a smooth surface and hairy bee thorax. We found that the coiled fiber increases the work to fracture of the entire cribellate thread by up to 20-fold. We also found that the axial fiber breaks multiple times during deformation, an unexpected observation that indicates: (i) the axial fiber continues to contribute work even after breakage, and (ii) the cribellate nanofibrils may perform a previously unidentified role as a binder material that distributes forces throughout the thread. Work of adhesion increased on surfaces with more surface structures (hairy bee thorax) corresponding to increased deformation of the coiled fiber. Together, our observations highlight how the synergistic interactions among the constituents of this natural composite adhesive enhance functionality. These highly extensible threads may serve to expose additional cribellate nanofibrils to form attachment points with prey substrata while also immobilizing prey as they sink into the web due to gravity.

Nitrogen inaccessibility protects spider silk from bacterial growth.

Spider silks are protein-based fibers that are incorporated into webs with the unique combination of high mechanical toughness and resistance to microbial degradation. While spiders are undoubtedly exposed to saprophytic microorganisms in their native habitats, such as the forest understory and bush, their silks have rarely been observed to decompose in either field or laboratory studies. We performed cross-streaking assays using silk from three spider species and four bacterial strains and found no inhibition zones, indicating the absence of antibacterial properties. We also cultured all bacteria directly upon silk in Luria-Bertani (LB) broth (full nutrients), phosphate-buffered saline (PBS; no nutrients) and nitrogen-free glucose broth (NFG; full nutrients, no nitrogen), and found that bacteria grew readily on silk in LB broth but not in PBS or NFG buffer. Our results indicate that spider silk's resistance to bacterial degradation is likely due to bacteriostatic rather than antibacterial mechanisms when nitrogen is inaccessible.

Filamentous calcareous alga provides substrate for coral-competitive macroalgae in the degraded lagoon of Dongsha Atoll, Taiwan.

BACKGROUND: The chemically-rich seaweed Galaxaura is not only highly competitive with corals, but also provides substrate for other macroalgae. Its ecology and associated epiphytes remain largely unexplored. To fill this knowledge gap, we undertook an ecological assessment to explore the spatial variation, temporal dynamics, and diversity of epiphytic macroalgae of Galaxaura divaricata on patch reefs in the lagoon of Dongsha Atoll, a shallow coral reef ecosystem in the northern South China Sea that has been repeatedly impacted by mass coral bleaching events. METHODS: Twelve spatially independent patch reefs in the Dongsha lagoon were first surveyed to assess benthic composition in April 2016, and then revisited to determine G. divaricata cover in September 2017, with one additional Galaxaura-dominated reef (site 9). Four surveys over a period of 17 months were then carried out on a degraded patch reef site to assess the temporal variation in G. divaricata cover. Epiphytic macroalgae associated with G. divaricata were quantified and identified through the aid of DNA barcoding at this degraded site. RESULTS: Patch reefs in the Dongsha lagoon were degraded, exhibiting relatively low coral cover (5-43%), but high proportions of macroalgae (13-58%) and other substrate (rubble and dead corals; 23-69%). The distribution of G. divaricata was heterogeneous across the lagoon, with highest abundance (16-41%) in the southeast area. Temporal surveys showed consistently high covers (mean ± SD = 16.9 ± 1.21%) of G. divaricata for 17 months. Additional photographic evidence suggested that overgrowth of G. divaricata can persist for 3.5 years. Yet, G. divaricata provides substrate to other macroalgae (e.g., Lobophora sp.) that also limit the growth of corals. CONCLUSIONS: Our study demonstrates that an allelopathic seaweed, such as G. divaricata, can overgrow degraded coral reefs for extended periods of time. By providing habitat for other harmful macroalgae, a prolonged Galaxaura overgrowth could further enhance the spread of macroalgae, and strengthen negative feedback loops, decreasing the recovery potential of degraded reefs.

When males live longer: Resource-driven territorial behavior drives sex-specific survival in snakes.

Phylogenetic analysis has shown that males' propensity to engage in aggressive encounters is associated with females having greater longevity. Here, we confirm the causal link between aggression and reduced longevity by looking at an egg-eating snake (Oligodon formosanus) in which females defend territories in the presence of sea turtle eggs. We monitored aggressiveness and survival at two sites: a control site with a stable supply of turtle eggs, and a second site where we collected data before and after a storm that eroded the beach on which turtles nested, thus leading to a loss of territoriality. We show that territoriality was the driver behind higher injury rates in females. Territorial females also had lower survival and decreased longevity compared with the nonterritorial males, but these differences disappeared when females were not territorial. Our study demonstrates how resource availability can influence the evolution of sex-specific patterns of survival across vertebrates.

Rafting on floating fruit is effective for oceanic dispersal of flightless weevils.

Terrestrial species, especially non-vagile ones (those unable to fly or swim), cannot cross oceans without exploiting other animals or floating objects. However, the colonisation history of flightless Pachyrhynchus weevils, inferred from genetic data, reveals their ability to travel long distances to colonise remote islands. Here, we used captive-bred Pachyrhynchus jitanasaius to analyse (i) the physiological tolerance of weevils (egg, larva and adult stages) to different levels of salinity; (ii) the survival rate of larvae in a simulated ocean environment in the laboratory; and (iii) the survival rate of larvae in a field experiment in the ocean using fruit of the fish poison tree floating on the Kuroshio current in the Pacific Ocean. We found that the survival rate of larvae in seawater was lower than in fresh water, although if the larvae survived 7 days of immersion in seawater, some emerged as adults in the subsequent rearing process. No adults survived for more than 2 days, regardless of salinity level. After floating separately for 6 days in salt water in the laboratory and in the Kuroshio current, two of 18 larvae survived in the fruit. This study provides the first empirical evidence that P. jitanasaius larvae can survive 'rafting' on ocean currents and that the eggs and larvae of these weevils have the highest probability of crossing the oceanic barrier. This ability may facilitate over-the-sea dispersal of these flightless insects and further shape their distribution and speciation pattern in the Western Pacific islands.

The effects of contemporary selection and dispersal limitation on the community assembly of acidophilic microalgae.

Extremophilic microalgae are primary producers in acidic habitats, such as volcanic sites and acid mine drainages, and play a central role in biogeochemical cycles. Yet, basic knowledge about their species composition and community assembly is lacking. Here, we begin to fill this knowledge gap by performing the first large-scale survey of microalgal diversity in acidic geothermal sites across the West Pacific Island Chain. We collected 72 environmental samples in 12 geothermal sites, measured temperature and pH, and performed rbcL amplicon-based 454 pyrosequencing. Using these data, we estimated the diversity of microalgal species, and then examined the relative contribution of contemporary selection (i.e., local environmental variables) and dispersal limitation on the assembly of these communities. A species delimitation analysis uncovered seven major microalgae (four red, two green, and one diatom) and higher species diversity than previously appreciated. A distance-based redundancy analysis with variation partitioning revealed that dispersal limitation has a greater influence on the community assembly of microalgae than contemporary selection. Consistent with this finding, community similarity among the sampled sites decayed more quickly over geographical distance than differences in environmental factors. Our work paves the way for future studies to understand the ecology and biogeography of microalgae in extreme habitats.

Nutrient intake determines post-maturity molting in the golden orb-web spider Nephila pilipes (Araneae: Araneidae).

While molting occurs in the development of many animals, especially arthropods, post-maturity molting (PMM, organisms continue to molt after sexual maturity) has received little attention. The mechanism of molting has been studied intensively; however, the mechanism of PMM remains unknown although it is suggested to be crucial for the development of body size. In this study, we investigated factors that potentially induce PMM in the golden orb-web spider Nephila pilipes, which has the greatest degree of sexual dimorphism among terrestrial animals. We manipulated the mating history and the nutrient consumption of the females to examine whether they affect PMM. The results showed that female spiders under low nutrition were more likely to molt as adults, and mating had no significant influence on the occurrence of PMM. Moreover, spiders that underwent PMM lived longer than those that did not and their body sizes were significantly increased. Therefore, we concluded that nutritional condition rather than mating history affect PMM.

The evolution of genital complexity and mating rates in sexually size dimorphic spiders.

BACKGROUND: Genital diversity may arise through sexual conflict over polyandry, where male genital features function to manipulate female mating frequency against her interest. Correlated genital evolution across animal groups is consistent with this view, but a link between genital complexity and mating rates remains to be established. In sexually size dimorphic spiders, golden orbweaving spiders (Nephilidae) males mutilate their genitals to form genital plugs, but these plugs do not always prevent female polyandry. In a comparative framework, we test whether male and female genital complexity coevolve, and how these morphologies, as well as sexual cannibalism, relate to the evolution of mating systems. RESULTS: Using a combination of comparative tests, we show that male genital complexity negatively correlates with female mating rates, and that levels of sexual cannibalism negatively correlate with male mating rates. We also confirm a positive correlation between male and female genital complexity. The macroevolutionary trajectory is consistent with a repeated evolution from polyandry to monandry coinciding with the evolution towards more complex male genitals. CONCLUSIONS: These results are consistent with the predictions from sexual conflict theory, although sexual conflict may not be the only mechanism responsible for the evolution of genital complexity and mating systems. Nevertheless, our comparative evidence suggests that in golden orbweavers, male genital complexity limits female mating rates, and sexual cannibalism by females coincides with monogyny.

Crypsis via leg clustering: twig masquerading in a spider.

The role of background matching in camouflage has been extensively studied. However, contour modification has received far less attention, especially in twig-mimicking species. Here, we studied this deceptive strategy by revealing a special masquerade tactic, in which the animals protract and cluster their legs linearly in the same axis with their bodies when resting, using the spider Ariamnes cylindrogaster as a model. We used cardboard papers to construct dummies resembling spiders in appearance and colour. To differentiate the most important factors in the concealment effect, we manipulated body size (long or short abdomen) and resting postures (leg clustered or spread) of the dummies and recorded the responses of predators to different dummy types in the field. The results showed that dummies with clustered legs received significantly less attention from predators, regardless of the body length. Thus, we conclude that A. cylindrogaster relies on the resting posture rather than body size for predator avoidance. This study provides, to the best of our knowledge, empirical evidence for the first time that twig-mimicking species can achieve effective camouflage by contour modification.