Investigation of potential gene flow limitation of behavioral adaptation in an aridlands spider

Summary This study investigates the possibility that gene flow underlies the apparently maladaptive behavior of a riparian woodland population of the desert spider Agelenopsis aperta with respect to territorial, foraging, and antipredatory behaviors. I found that other local populations of A. aperta in the vicinity of the riparian woodland habitat are prey-limited and exhibit an aridlands phenotype (high aggressiveness in competitive interactions over energy-based territories and a lack of discrimination among potential prey types). The riparian woodland population deviates from surrounding populations in the area in that prey are abundant and this population shows a mixture of aridlands and riparian (low aggressiveness towards conspecifics and discrimination of prey profitability) phenotypes. Electrophoretic analyses of population subdivision in the area indicate that significant levels of gene flow have occurred, at least, sometime in the past. Drift fence analyses of spider movement futher indicate that there is marked unidirectional movement of spiders each year from the more arid habitats into the riparian woodland. Experimental manipulation of gene flow and predation pressure demonstrates that gene flow restricts adaptation in this habitat: one generation of predation pressure in the absence of gene flow is sufficient to cause a marked shift in spider behavior towards the expected riparian phenotype.     

Population differences in how black-tailed prairie dogs deal with snakes

Summary This paper examines the antisnake behavior of a snake-experienced Texas (TX) population and a snake-naive South Dakota (SD) population of black-tailed prairie dogs (Cynomys ludovicianus). Animals in both populations were presented with tethered rattlesnakes and nonvenomous bullsnakes before and after the first emergence of pups from their natal burrows. SD and TX adults were more likely to actively harass snakes than were pups. There were no differences in the behavior of SD adult males vs females or fathers vs non-fathers, contrary to what was found in TX. However, SD mothers did call more and stayed closer to snakes than did SD non-mothers. All SD adult sex-parental classes spent less time dealing with snakes after the emergence of pups, which was not observed in TX. The primary difference between the two populations was the behavior of TX males who spent a great deal of time harassing snakes. SD adults behaved most like TX females in that these groups spent little time actively dealing with snakes. Pups in both populations behaved similarly. These results are interpreted in terms of the relationship between potential predator and potential prey.     

Life history, ecology and the biogeography of strong genetic breaks among 15 species of Pacific rockfish, Sebastes

Strong genetic change over short spatial scales is surprising among marine species with high dispersal potential. Concordant breaks among several species signals a role for geographic barriers to dispersal. Along the coast of California, such breaks have not been seen across the biogeographic barrier of Point Conception, but other potential geographic boundaries have been surveyed less often. We tested for strong-population structure in 11 species of Sebastes sampled across two regions containing potential dispersal barriers, and conducted a meta-analysis including four additional species. We show two strong breaks north of Monterey Bay, spanning an oceanographic gradient and an upwelling jet. Moderate genetic structure is just as common in the north as it is in the south, across the biogeographic break at Point Conception. Gene flow is generally higher among deep-water species, but these conclusions are confounded by phylogeny. Species in the subgenus Sebastosomus have higher structure than those in the subgenus Pteropodus, despite having larvae with longer pelagic phases. Differences in settlement behavior in the face of ocean currents might help explain these differences. Across similar species across the same coastal environment, we document a wide variety of patterns in gene flow, suggesting that interaction of individual species traits such as settlement behavior with environmental factors such as oceanography can strongly impact population structure.     

Ecological Trap for Desert Lizards Caused by Anthropogenic Changes in Habitat Structure that Favor Predator Activity

Abstract: Anthropogenic habitat perturbation is a major cause of population decline. A standard practice managers use to protect populations is to leave portions of natural habitat intact. We describe a case study in which, despite the use of this practice, the critically endangered lizard Acanthodactylus beershebensis was locally extirpated from both manipulated and natural patches within a mosaic landscape of an afforestation project. We hypothesized that increased structural complexity in planted patches favors avian predator activity and makes these patches less suitable for lizards due to a heightened risk of predation. Spatial rarity of natural perches (e.g., trees) in arid scrublands may hinder the ability of desert lizards to associate perches with low‐quality habitat, turning planted patches into ecological traps for such species. We erected artificial trees in a structurally simple arid habitat (similar to the way trees were planted in the afforestation project) and compared lizard population dynamics in plots with these structures and without. Survival of lizards in the plots with artificial trees was lower than survival in plots without artificial trees. Hatchlings dispersed into plots with artificial trees in a manner that indicated they perceived the quality of these plots as similar to the surrounding, unmanipulated landscape. Our results showed that local anthropogenic changes in habitat structure that seem relatively harmless may have a considerable negative effect beyond the immediate area of the perturbation because the disturbed habitat may become an ecological trap.     

Behavioural determinants of gene flow in the bat Miniopterus schreibersii

Dispersal and migratory behaviours are often important determinants of gene flow in wild species, and we have studied their role using ringing-recapture data in the Portuguese population of Miniopterus schreibersii, a cave-dwelling bat that forms large maternity colonies. Juvenile dispersal, usually a major agent of gene flow, appears to be negligible, as young females never settled to give birth in foreign colonies. Likewise, there was virtually no dispersal of adult females to foreign maternity colonies. This strong philopatry virtually eliminated female-mediated gene flow, but we found a great potential for male-mediated gene flow among colonies, as regional migrations temporarily joined both sexes from different colonies in the same roosts, during the mating season. In fact, females from some colonies were more likely to mate with males from foreign colonies than from their own, thus potentially bringing home genes of foreign males. In spite of this abundant gene flow, we found a pattern of isolation by distance and even strong barriers to gene flow, which is explained by the fact that migrations were usually too short to allow direct flow among distant colonies. We concluded that potential gene flow is influenced by the distance between colonies and the availability of mating roosts between them. In addition, we found this flow to be asymmetrical, with a dominant direction from the largest to the smallest colonies. Our ringing-recapture estimates of potential gene flow based on dispersal and migratory behaviour are compatible with the genetic structure of the population for both nuclear and mitochondrial DNA. Our conclusions have implications for the conservation of bats with a spatial behaviour similar to that of M. schreibersii. Colonies should be managed individually because strict female philopatry not only promotes their isolation but also minimises the contribution of immigration in rescuing declining colonies. Furthermore, the results underline the importance of preserving mating roosts to maintain gene flow among colonies.     

Within-group behavioral variation promotes biased task performance and the emergence of a defensive caste in a social spider

The social spider Anelosimus studiosus exhibits a behavioral polymorphism where colony members express either a passive, tolerant behavioral tendency (social) or an aggressive, intolerant behavioral tendency (asocial). Here we test whether asocial individuals act as colony defenders by deflecting the suite of foreign (i.e., heterospecific) spider species that commonly exploit multi-female colonies. We (1) determined whether the phenotypic composition of colonies is associated with foreign spider abundance, (2) tested whether heterospecific spider abundance and diversity affect colony survival in the field, and (3) performed staged encounters between groups of A. studiosus and their colony-level predator Agelenopsis emertoni (A. emertoni)to determine whether asocial females exhibit more defensive behavior. We found that larger colonies harbor more foreign spiders, and the number of asocial colony members was negatively associated with foreign spider abundance. Additionally, colony persistence was negatively associated with the abundance and diversity of foreign spiders within colonies. In encounters with a colony-level predator, asocial females were more likely to exhibit escalatory behavior, and this might explain the negative association between the frequency of asocial females and the presence of foreign spider associates. Together, our results indicate that foreign spiders are detrimental to colony survival, and that asocial females play a defensive role in multi-female colonies.     

A complex social structure with fission–fusion properties can emerge from a simple foraging model

Precisely how ecological factors influence animal social structure is far from clear. We explore this question using an agent-based model inspired by the fission–fusion society of spider monkeys (Ateles spp). Our model introduces a realistic, complex foraging environment composed of many resource patches with size varying as an inverse power law frequency distribution with exponent β. Foragers do not interact among them and start from random initial locations. They have either a complete or a partial knowledge of the environment and maximize the ratio between the size of the next visited patch and the distance traveled to it, ignoring previously visited patches. At intermediate values of β, when large patches are neither too scarce nor too abundant, foragers form groups (coincide at the same patch) with a similar size frequency distribution as the spider monkey’s subgroups. Fission–fusion events create a network of associations that contains weak bonds among foragers that meet only rarely and strong bonds among those that repeat associations more frequently than would be expected by chance. The latter form subnetworks with the highest number of bonds and a high clustering coefficient at intermediate values of β. The weak bonds enable the whole social network to percolate. Some of our results are similar to those found in long-term field studies of spider monkeys and other fission–fusion species. We conclude that hypotheses about the ecological causes of fission–fusion and the origin of complex social structures should consider the heterogeneity and complexity of the environment in which social animals live.     

干旱区的城市生境特征与绿洲化对策

干旱地区的城市亦被称为绿洲。从其发生与生境状况分析,注意到近期城市的扩展已脱离绿洲型的发展轨道,而与全球城市模式类同,亦具城市化的共有弊病。又因所处区位受到荒漠气候的强烈影响,较其它地带的城市,其生境更加恶劣。干旱区居民长期凭借绿洲生境而生息劳动,早期出现的城市亦脱胎于绿洲。目前仍沿用绿洲以称城市,但与绿洲的生境相比较已面目全非,实质是以现代设施在荒漠中建立的水泥荒漠。目前全球范围正展开以改善城市生境为目标的生态城市建设,位处干旱区的城市应从其所在区位出发,以绿洲化作为生态城市建设目标,切实改善居民生存环境质量。应仿照绿洲,在外围营造阻隔荒漠气候侵袭的防护林,在市区通过营造片林与行道树,形成类似条田林网格局,以一定植被规模重构绿洲化城市,形成绿洲生境以改善城市居民生存条件,回归干旱区居民世世代代依存的绿洲空间。     

Potential Effects of the United States-Mexico Border Fence on Wildlife

Abstract:  Security infrastructure along international boundaries threatens to degrade connectivity for wildlife. To explore potential effects of a fence under construction along the U.S.–Mexico border on wildlife, we assessed movement behavior of two species with different life histories whose regional persistence may depend on transboundary movements. We used radiotelemetry to assess how vegetation and landscape structure affect flight and natal dispersal behaviors of Ferruginous Pygmy-Owls ( Glaucidium brasilianum ), and satellite telemetry, gene-flow estimates, and least-cost path models to assess movement behavior and interpopulation connectivity of desert bighorn sheep ( Ovis canadensis mexicana ). Flight height of Pygmy-Owls averaged only 1.4 m (SE 0.1) above ground, and only 23% of flights exceeded 4 m. Juvenile Pygmy-Owls dispersed at slower speeds, changed direction more, and had lower colonization success in landscapes with larger vegetation openings or higher levels of disturbance ( p ≤ 0.047), which suggests large vegetation gaps coupled with tall fences may limit transboundary movements. Female bighorn sheep crossed valleys up to 4.9 km wide, and microsatellite analyses indicated relatively high levels of gene flow and migration (95% CI for F_ST= 0.010–0.115, Nm = 1.9–24.8, M = 10.4–15.4) between populations divided by an 11-km valley. Models of gene flow based on regional topography and movement barriers suggested that nine populations of bighorn sheep in northwestern Sonora are linked by dispersal with those in neighboring Arizona. Disruption of transboundary movement corridors by impermeable fencing would isolate some populations on the Arizona side. Connectivity for other species with similar movement abilities and spatial distributions may be affected by border development, yet mitigation strategies could address needs of wildlife and humans.     

Genetic diversity and population structure of the Chinese mitten crab <Emphasis Type="Italic">Eriocheir sinensis</Emphasis> in its native range

The Chinese mitten crab Eriocheir sinensis is an indigenous and economically important species in China, but can also be found as invasive species in Europe and America. Mitten crabs have been exploited extensively as a food resource since the 1990s. Despite its ecological and economic importance, the genetic structure of native mitten crab populations is not well understood. In this paper, we investigated the genetic structure of mitten crab populations in China by screening samples from ten locations covering six river systems at six microsatellite loci. Our results provide further evidence that mitten crabs from the River Nanliujiang in Southern China are a genetically differentiated population within the native range of Eriocheir, and should be recognized as a separate taxonomic unit. In contrast, extremely low levels of genetic differentiation and no significant geographic population structure were found among the samples located north of the River Nanliujiang. Based on the reproductive biology of mitten crabs and the geography of their habitat we argue that both natural and human-mediated gene flow are unlikely to fully account for the similar allele frequency distributions at microsatellite loci. Large population sizes of mitten crabs suggest instead that a virtual absence of genetic drift and significant homoplasy of microsatellite alleles have contributed to the observed pattern. Furthermore, a coalescent-based maximum likelihood method indicated a more than two-fold lower effective population size of the Southern population compared to the Northern Group and low but significant levels of gene flow between both areas.     

Genetic structure across the GBR: evidence from short-lived gobies

The Great Barrier Reef (GBR) exhibits distinct cross-shelf zonation. These patterns are particularly well documented in reef fishes and have been attributed to either environmental gradients (e.g. wave energy, oceanography) or barriers to gene flow. This study examined the extent to which barriers to gene flow contribute to cross-shelf patterns by examining the mitochondrial DNA of gobies (genus Eviota). The genus Eviota was selected due to its extreme life history characteristics (shortest vertebrate lifespan) and cross-shelf distribution patterns (E. queenslandica, inner- and mid-shelf, and E. albolineata mid- and outer-shelf). Although cross-shelf barriers to gene flow were predicted, this study found no population structure between shelf locations. However, a genetically distinct population of E. queenslandica (the inner-shelf species) was observed at North Direction Island (Phi_st = 0.088, P = 0.004). As no comparable structure was observed in E. albolineata (the outer-shelf species) it may be that habitat type (E. queenslandica = reef lagoon, E. albolineata = reef crest) is a significant factor driving the structure observed in E. queenslandica. Larval behaviour, olfactory or auditory senses and reef selection at settlement could be assisting larvae to return to reefs similar to natal reefs. We suggest that ecological gradients are more important than barriers to gene flow in structuring cross-shelf distributions within Eviota.     

宁夏地区二斑叶螨的寄主植物选择及其季节转移

采用田间系统观察和室内饲养方法,调查并发现宁夏引黄灌区和新垦区二斑叶螨（Tetranychus urticae Koch)为寄主植物88种,其中野生寄主植物59种,栽培寄主29种,垦前荒漠,半荒漠有适宜二斑叶螨生存和繁殖的寄主植物分布,用生命表方法研究了20种主要寄主植物对二斑叶螨实验种群净生殖率（R0)的营养效应,不同寄主植物随季节的变化其营养效应明显不同,对二斑叶螨在这些寄主植物间的季节转移规律研究表明,宁夏农田,以野荠菜Capsella bursa-pastoris,小旋花Calystegia hederacea,小蓟Cephalanoplos segetumji,苦苣菜Sonchus brachyotus,车前草Plantago asiatica,蚕豆Vicia foba,大豆Glycine max,玉米Zea mays等一起,构成了有利于二斑叶螨季节转移的寄主体系,分析认为,宁夏麦套玉米间作大豆的耕种模式是二斑叶螨大发生的主要原因之一。     

Song dialects do not restrict gene flow in an urban population of the orange-tufted sunbird, <Emphasis Type="Italic">Nectarinia osea</Emphasis>

Geographic variation in vocalizations is widespread in passerine birds, but its origins and maintenance remain unclear. In this study, we test the hypothesis that song dialect, a culturally transmitted trait, is related to the population genetic structure of the orange-tufted sunbird, Nectarinia osea. To address this, we compared mitochondrial DNA (mtDNA) sequence variation together with allele frequencies at five microsatellite loci from an urban population of sunbirds exhibiting two distinct song dialects on a microgeographic scale. Our findings reveal no association between dialect membership and genetic composition. All genetic measures, from both mitochondrial and nuclear DNA, indicate high levels of gene flow between both dialect populations. The low F _ST values obtained from mtDNA and microsatellite analysis imply that the variation among dialects does not account for more than 2%, at best, of the overall genetic variation found in the entire population. These measures fall well within the range of similar measures obtained in other studies of species exhibiting vocal dialects, most of which fail to detect any dialect-based genetic differentiation. The persistence of dialects in the orange-tufted sunbird may thus best be explained by dispersal of individuals across dialect boundaries and possibly from surrounding areas, followed by postdispersal vocal matching. Because genetic structuring appears weaker than cultural structure in this species, we discuss the behavioral mechanisms underlying dialect maintenance in the presence of apparent gene flow.     

Recognising one’s enemies: a functional approach to risk assessment by prey

Little has been done to compare the relative importance of various mechanisms through which prey assess the potential risk from natural enemies. We used predator-naive spider mites (Tetranychus urticae, Tetranychidae) to (1) compare the responses of prey to chemical cues from enemy and non-enemy species and (2) investigate the source of these cues. In the laboratory, we observed the distribution of T. urticae in response to cues from nine mite species, including (1) predators of spider mites, (2) predators/parasites of other animals, and (3) fungivores/pollen-feeders. When given a choice over 24 h, spider mites foraged and oviposited in fewer numbers on leaf discs that were previously exposed to predatory or parasitic mites (including species incapable of attacking spider mites) than on clean leaf discs (unexposed to mites). Interestingly, previous exposure of leaf arenas to fungivores and pollen-feeders had no significant effect on spider mite distribution. We then observed the response of T. urticae to cues from two species of predator that had been reared on a diet of either spider mites or pollen. T. urticae showed stronger avoidance of leaf discs that were previously exposed to spider-mite-fed predators than of discs exposed to pollen-fed predators. Nevertheless, for one predator species (Amblyseius andersoni), T. urticae still preferred to forage and oviposit on clean (unexposed) discs than on discs exposed to pollen-fed predators. Protein-derived metabolic wastes of predatory or parasitic mites may provide a general cue about potential predation risk for T. urticae. However, T. urticae also avoided areas exposed to pollen-fed predators, suggesting there may be other sources of enemy recognition by the spider mites. We discuss the ecological and evolutionary mechanisms that may influence the scope of information through which animals assess predation risk. Received: 11 January 1999 / Received in revised form: 25 October 1999 / Accepted: 20 November 1999     

Environmental impact propagated by cross-system subsidy: chronic stream pollution controls riparian spider populations

Resource subsidies between habitats are common and create the potential for the propagation of environmental impacts across system boundaries. However, recent understanding of the potential for subsidy-mediated cross-system impact propagations is limited and primarily based on passive flows of nutrients and detritus or short-term effects. Here, we assess the effects of sustained alterations in aquatic insect emergence (active subsidy pathway), due to chronic stream pollution, for riparian spiders. The sustained reduction in aquatic insect densities at the polluted reaches resulted in a marked decline in web spider population density and a shift in spider community composition. Our results provide the first evidence that stream pollution can control populations and community structure of terrestrial predators via sustained alterations in aquatic subsidies, emphasizing the role of subtle trophic linkages in the transmission of environmental impacts across ecosystem boundaries.     

Behavior of aqueous stable colloidal nano-C60 aggregates exposed to TX100 micelles under different environmental conditions

C₆₀, as one of carbon nanomaterials widely used in various fields, could be released into the water environment thus exerting some potential health risks to human beings. This work examined the behavior of aqueous stable colloidal nano-C₆₀ (nC₆₀) aggregates under different environmental conditions including Polyethylene glycol octylphenol ether (TX100) micelles concentration, pH, and reaction time when exposed to TX100 micelles. Results show that the nC₆₀ aggregates became more dispersive and restored the capability of generating the singlet oxygen when exposed to TX100 micelles. With the increase of TX100 concentration, smaller average size of nC₆₀ aggregates was observed in dynamic light scattering (DLS) analysis, the fluorescence intensity of TX100 was more quenched by nC₆₀ aggregates, and the kinetic rate constant of generating the singlet oxygen for nC₆₀ aggregates was improved. The mean size of nC₆₀ aggregates in the presence of TX100 had no obvious variations when the pH ranged from 4 to 8. The longer reaction time between nC₆₀ aggregates and TX100 led to a higher kinetic rate constant of generating the singlet oxygen. Collective data suggest that variations in physicochemical properties of nC₆₀ aggregates are strongly dependent on the surrounding media under different environmental conditions and directly govern nC₆₀’s transport behavior and potential toxicity.     

Predator foraging behavior in response to perception and learning by its prey: interactions between orb-spinning spiders and stingless bees

Although rewarded bees learn and remember colors and patterns, they have difficulty in learning to avoid negative stimuli such as decorated spider webs spun by Argiope argentata. A. argentata decorates its web with silk patterns that vary unpredictably (Fig. 1) and thus foraging insects that return to sites where spiders are found encounter new visual cues daily. Stingless bees can learn to avoid spider webs but avoidance-learning is slowed or inhibited by daily variation in web decorations (Figs. 3,4; Tables 1,2). In addition, even if bees learn to avoid decorated webs found in one location, they are unable to generalize learned-avoidance responses to similarly decorated webs found at other sites. A. argentata seems to have evolved a foraging behavior that is tied to the ways insects perceive and process information about their environment. Because of the evolutionary importance of bee-flower interdependence, the predatory behavior of web-decorating spiders may be difficult for natural selection to act against.     

Postfire Logging in Riparian Areas

Abstract:  We reviewed the behavior of wildfire in riparian zones, primarily in the western United States, and the potential ecological consequences of postfire logging. Fire behavior in riparian zones is complex, but many aquatic and riparian organisms exhibit a suite of adaptations that allow relatively rapid recovery after fire. Unless constrained by other factors, fish tend to rebound relatively quickly, usually within a decade after a wildfire. Additionally, fire and subsequent erosion events contribute wood and coarse sediment that can create and maintain productive aquatic habitats over time. The potential effects of postfire logging in riparian areas depend on the landscape context and disturbance history of a site; however, available evidence suggests two key management implications: (1) fire in riparian areas creates conditions that may not require intervention to sustain the long-term productivity of the aquatic network and (2) protection of burned riparian areas gives priority to what is left rather than what is removed. Research is needed to determine how postfire logging in riparian areas has affected the spread of invasive species and the vulnerability of upland forests to insect and disease outbreaks and how postfire logging will affect the frequency and behavior of future fires. The effectiveness of using postfire logging to restore desired riparian structure and function is therefore unproven, but such projects are gaining interest with the departure of forest conditions from those that existed prior to timber harvest, fire suppression, and climate change. In the absence of reliable information about the potential consequence of postfire timber harvest, we conclude that providing postfire riparian zones with the same environmental protections they received before they burned is justified ecologically. Without a commitment to monitor management experiments, the effects of postfire riparian logging will remain unknown and highly contentious.     

Community composition and nearest neighbor relationships in orb-weaving spiders: the product of aggression?

Summary Nearest-neighbor distances and species' densities were calculated for 949 individuals of 8 species of orb-weaving spiders from eight edge and meacow sites in Panama. These data were used to test the hypothesis that interspecific aggressive interactions influenced spider community structure by: (1) eliminating or decreasing the densities of sympatric, less aggressive species, or, (2) affecting nearest-neighbor relationships so that large or aggressive individuals had smaller, less aggressive or more distant neighbors. Although there was a negative relationship between the densities of two species pairs (Leucauge sp. and Mangora pia; Eriophora fuliginea and Cyclosa caroli), overall spider densities could not be predicted from indices of spider aggression obtained by introduction experiments. The spatial distribution of spiders in three dimensions was not significantly different than random. I attributed the lack of clumping to the relative vegetational homogeneity of my sites; the lack of overdispersion may reflect the fact that websites are not limiting. Relationships between nearest neighbors generally supported the aggression hypothesis, however, comparisons of the actual data with equivalent randomly-assorted data indicate that a hypothesis of randomly chosen neighborsis equally applicable. This result suggests that, unless websites differ significantly in their quality, with high quality websites in short supply, spider aggression may not affect community structure.     

Social spider defense against kleptoparasitism

Summary Because of the large amount of webbing they provide, social spider colonies often host other satellite spider species referred to as kleptoparasites or food stealers. Such kleptoparasites may take advantage of increased prey capture rates associated with large spider aggregations. This study investigated the relationship between a cooperatively social spider species, Anelosimus eximius (Araneae: Theridiidae), which lives in the undergrowth of tropical rainforests in Peru, and its kleptoparasite, Argyrodes ululans (Araneae: Theridiidae), which specializes in foraging in An. eximius webs. Although large aggregates of spiders may be more attractive to kleptoparasites, the benefits of group defense may offset this cost. Natural colonies were observed, and enclosed field colonies containing fixed numbers of host spiders were manipulated in order to determine whether kleptoparasite success is affected by the number of social spiders that are available for defense. Prey was less likely to be stolen by Ar. ululans when a greater number of host An. eximius spiders were involved in prey capture. When hosts detected a kleptoparasite earlier and chased it more often, prey was more likely to be successfully defended. Ar. ululans was more successful in stealing small prey items in all colonies and gave up more readily on very large prey (> 11 mm). I conclude that communal living and group defense in An. eximius confer protection from the kleptoparasite Ar. ululans.