Endosymbiont diversity across native and invasive brown widow spider populations.

The invasive brown widow spider, Latrodectus geometricus (Araneae: Theridiidae), has spread in multiple locations around the world and, along with it, brought associated organisms such as endosymbionts. We investigated endosymbiont diversity and prevalence across putative native and invasive populations of this spider, predicting lower endosymbiont diversity across the invasive range compared to the native range. First, we characterized the microbial community in the putative native (South Africa) and invasive (Israel and the United States) ranges via high throughput 16S sequencing of 103 adult females. All specimens were dominated by reads from only 1-3 amplicon sequence variants (ASV), and most individuals were infected with an apparently uniform strain of Rhabdochlamydia. We also found Rhabdochlamydia in spider eggs, indicating that it is a maternally-inherited endosymbiont. Relatively few other ASV were detected, but included two variant Rhabdochlamydia strains and several Wolbachia, Spiroplasma and Enterobacteriaceae strains. We then diagnostically screened 118 adult female spiders from native and invasive populations specifically for Rhabdochlamydia and Wolbachia. We found Rhabdochlamydia in 86% of individuals and represented in all populations, which suggests that it is a consistent and potentially important associate of L. geometricus. Wolbachia was found at lower overall prevalence (14%) and was represented in all countries, but not all populations. In addition, we found evidence for geographic variation in endosymbiont prevalence: spiders from Israel were more likely to carry Rhabdochlamydia than those from the US and South Africa, and Wolbachia was geographically clustered in both Israel and South Africa. Characterizing endosymbiont prevalence and diversity is a first step in understanding their function inside the host and may shed light on the process of spread and population variability in cosmopolitan invasive species.

Why do predators attack parasitized prey? Insights from a probabilistic model and a literature survey.

Predators and parasitoids often encounter parasitized prey or hosts during foraging. While the outcomes of such encounters have been extensively studied for insect parasitoids, the consequences of a predator encountering parasitized prey have received less attention. One extreme example involves the potter wasp Delta dimidiatipenne that frequently provision their nest with parasitized caterpillars, despite the low suitability of this prey for consumption by their offspring. This raises two main questions: (1) why do female potter wasps continue collecting parasitized caterpillars? and (2) is this an exceptional example, or do predatory insects often suffer from fitness costs due to encounters with parasitized prey? We addressed the first question using a probabilistic mathematical model predicting the value of discrimination between parasitized and unparasitized prey for the potter wasp, and the second question by surveying the literature for examples in which the parasitism status of prey affected prey susceptibility, suitability, or prey choice by a predator. The model demonstrates that only under certain conditions is discrimination against parasitized prey beneficial in terms of the potter wasp's lifetime reproductive success. The literature survey suggests that the occurrence of encounters and consumption of parasitized prey is common, but the overall consequences of such interactions have rarely been quantified. We conclude that the profitability and ability of a predator to discriminate against parasitized prey under natural conditions may be limited and call for additional studies quantifying the outcome of such interactions.

Trait-based approaches to predicting biological control success: challenges and prospects.

Identifying traits that are associated with success of introduced natural enemies in establishing and controlling pest insects has occupied researchers and biological control practitioners for decades. Unfortunately, consistent general relationships have been difficult to detect, preventing a priori ranking of candidate biological control agents based on their traits. We summarise previous efforts and propose a series of potential explanations for the lack of clear patterns. We argue that the quality of current datasets is insufficient to detect complex trait-efficacy relationships and suggest several measures by which current limitations may be overcome. We conclude that efforts to address this elusive issue have not yet been exhausted and that further explorations are likely to be worthwhile.

Invasive brown widow spiders avoid parasitism despite high densities.

Invasive species are sometimes less susceptible to natural enemies compared to native species, but the mechanism is often unclear. Here we tested two potential mechanisms for lower parasitism of invasive species: density-dependent parasitism and preference for human-dominated habitats. We investigated how variation in host density and habitat type affect egg sac parasitism in two widow spider species (family Theridiidae). We compared parasitism on the egg sac of the brown widow, Latrodectus geometricus, an urban invasive species, and the white widow, Latrodectus pallidus, a species native to Israel. To investigate variation in host and parasitoid density, we measured nearest-neighbor distance between spider webs and parasitism rates in 16 sites, and in a single site monthly throughout a year. In L. pallidus, denser sites were more heavily parasitized (up to 55%) and parasitism rate increased with population density throughout the season. Extremely dense L. geometricus populations, however, had very low rates of parasitism (0-5%). We then conducted an egg sac transplant experiment in human-dominated and natural habitats. We found no parasitism of either species in the human-dominated habitat, compared to 30% parasitism of both species in the natural habitat. In addition, we found evidence for higher predation of L. pallidus than of L. geometricus egg sacs, particularly in the natural habitat. These combined results suggest that the human-dominated habitats inhabited by L. geometricus have a lower abundance of predators and parasites. We conclude that lower parasitism and predation in human-dominated habitats could contribute to the invasion success of L. geometricus.

Invertebrate and vertebrate predation rates in a hyperarid ecosystem following an oil spill.

Extreme temperatures and scarce precipitation in deserts have led to abiotic factors often being regarded as more important than biotic ones in shaping desert communities. The presumed low biological activity of deserts is also one reason why deserts are often overlooked by conservation programs. We provide the first quantification of predation intensity from a desert ecosystem using artificial sentinel prey emulating caterpillars, a standardized monitoring tool to quantify relative predation pressure by many invertebrate and vertebrate predators. The study was conducted in a protected natural area affected by oil spills in 1975 and 2014; hence, we assessed the potential effects of oil pollution on predation rates. We found that predation was mostly due to invertebrate rather than vertebrate predators, fluctuated throughout the year, was higher at the ground level than in the tree canopy, and was not negatively affected by the oil spills. The mean predation rate per day (12.9%) was within the range found in other ecosystems, suggesting that biotic interactions in deserts ought not to be neglected and that ecologists should adopt standardized tools to track ecological functions and allow for comparisons among ecosystems.

The Impact of Roads on the Redistribution of Plants and Associated Arthropods in a Hyper-Arid Ecosystem.

The construction of vehicular roads likely affects the distribution of natural resources. Although the effects of roads on different ecosystem aspects have been extensively studied, studies in arid and, particularly, in hyper-arid ecosystems are scarce. In drylands, where water is the main limiting factor, the effect of roads on the redistribution of water may have strong subsequent effects on the ecosystem, especially when roads cross natural water flow paths. To fill this knowledge gap, we studied the effects of a road that runs across a slope on the distribution of plants and animals in a hyper-arid environment. Changes in shrub cover, below and above the road, were quantified by remote sensing and image classification, while plant-associated arthropods were vacuum-sampled from shrub canopies and from open (inter-shrub) areas. We found that the spatial distribution of shrubs, a vital resource facilitating many other organisms, was affected by the road, with an increase in the shrub cover immediately above the road and a decrease below it. Arthropod abundance generally followed shrub cover, but the exact pattern depended on the specific group sampled. While some arthropod groups (e.g., aphids, parasitic wasps and barklice) thrived under the disturbed conditions above the road, other arthropod groups (e.g., mites and true bugs) were less abundant in the disturbed patches. Our results highlight the strong effects of human-made structures on the distribution of flora and fauna in arid ecosystems.

Wolbachia Endosymbionts of Fleas Occur in All Females but Rarely in Males and Do Not Show Evidence of Obligatory Relationships, Fitness Effects, or Sex-Distorting Manipulations.

The widespread temporal and spatial persistence of endosymbionts in arthropod host populations, despite potential conflicts with their hosts and fluctuating environmental conditions, is puzzling. Here, we disentangled three main mechanisms that are commonly proposed to explain such persistence, namely, obligatory relationships, in which the host is fully dependent on its endosymbiont, fitness advantages conferred by the endosymbiont, and reproductive manipulations imposed by the endosymbiont. Our model system reflects an extreme case, in which the Wolbachia endosymbiont persists in all female flea hosts but rarely in male ones. We cured fleas of both sexes of Wolbachia but found no indications for either lower reproduction, offspring survival, or a change in the offspring sex ratio, compared to Wolbacia-infected fleas. These results do not support any of the suggested mechanisms. We highlight future directions to advance our understanding of endosymbiont persistence in fleas, as well as in other model systems, with extreme sex-differences in endosymbiont persistence. Insights from such studies are predicted to shed light on the evolution and ecology of arthropod-endosymbiont interactions in nature.

The Impact of Terrestrial Oil Pollution on Parasitoid Wasps Associated With Vachellia (Fabales: Fabaceae) Trees in a Desert Ecosystem, Israel.

Oil is a major pollutant of the environment, and terrestrial oil spills frequently occur in desert areas. Although arthropods account for a large share of animal diversity, the effect of oil pollution on this group is rarely documented. We evaluated the effects of oil pollution on parasitoid wasps associated with Vachellia (formerly Acacia) tortilis (Forssk.) and Vachellia raddiana (Savi) trees in a hyper-arid desert that was affected by two major oil spills (in 1975 and 2014). We sampled the parasitoid populations between 2016 and 2018 in three sampling sites and compared their abundance, diversity, and community composition between oil-polluted and unpolluted trees. Parasitoid abundance in oil-polluted trees was lower in one of the sites affected by the recent oil spill, but not in the site affected by the 1975 oil spill. Oil-polluted trees supported lower parasitoid diversity than unpolluted trees in some sampling site/year combinations; however, such negative effects were inconsistent and pollution explained a small proportion of the variation in parasitoid community composition. Our results indicate that oil pollution may negatively affect parasitoid abundances and diversity, although the magnitude of the effect depends on the tree species, sampling site, and the time since the oil spill.

Seed Predation on Oil-Polluted and Unpolluted Vachellia (Acacia) Trees in a Hyper-Arid Desert Ecosystem.

Acacia trees are keystone species in many arid environments, supporting high levels of plant and animal diversity. In Israel, the populations of Vachellia (formerly Acacia) tortilis (Forssk.) and V. raddiana (Savi) are declining at an alarming rate. Severe infestations by bruchid beetles (Coleoptera, Chrysomelidae) are among the major causes of seed mortality, but additional environmental stressors can reduce the defence level of the seeds, exacerbating their susceptibility to predators. In a hyper-arid desert ecosystem affected by two major oil spills (in 1975 and 2014), we quantified seed predation rates caused by insect granivores before and after the pods dropped to the ground. We recorded predation rates of up to 84% for both tree species, and higher predation rates at the ground level than in the canopy, suggesting that repeated infestations occur. These results reinforce the call to protect the populations of large ungulates such as gazelles, which kill the bruchids by feeding upon the pods, and promote seed germination and dispersion. We found no clear evidence of a negative effect of the oil spill on seed predation, indicating that oil pollution did not increase the vulnerability of the seeds to granivores even in trees affected by the recent oil spill.

Parasitoid Abundance and Community Composition in Desert Vineyards and Their Adjacent Natural Habitats.

Parasitoids are important natural enemies of many agricultural pests. Preserving natural habitats around agricultural fields may support parasitoid populations. However, the success of such an approach depends on the ability of parasitoids to utilize both crop and natural habitats. While these aspects have been studied extensively in temperate regions, very little is known about parasitoid communities in desert agroecosystems. We took one step in this direction by sampling parasitoids in six vineyards and their surrounding natural desert habitat in a hyper-arid region of the Negev Desert Highlands, Israel. We predicted that due to the high contrast in environmental conditions, parasitoid abundance and community composition would differ greatly between the crop and the natural desert habitats. We found that parasitoid abundance differed between the habitats; however, the exact distribution pattern depended on the time of year-with higher numbers of parasitoids in the natural habitat at the beginning of the vine growth season and higher numbers in the vineyard at the middle and end of the season. Although parasitoid community composition significantly differed between the vineyard and desert habitats, this only accounted for ~4% of the total variation. Overall, our results do not strongly support the notion of distinct parasitoid communities in the crop vs. the desert environment, suggesting that despite environmental contrasts, parasitoids may move between and utilize resources in both habitats.

Frequency and consequences of the collection of already parasitized caterpillars by a potter wasp.

Maladaptive behaviors reflecting a "bad" choice of habitat or resource have been widely documented; however, their persistence is often difficult to interpret. The potter wasp Delta dimidiatipenne constructs mud cells, in each of which it lays a single egg and places several caterpillars to feed its offspring. Preliminary observations indicated that a portion of these caterpillars were already parasitized and contained the offspring of the gregarious parasitoid Copidosoma primulum. As a result, the offspring of the potter wasp often failed to develop. To characterize the distribution, frequency and consequences of this intriguing phenomenon, we surveyed potter wasp nests throughout the Negev Desert. Evidence for parasitized caterpillars (mummies) was found in ~85% of the sampled sites, in ~20% of previous years' nest cells and in ~70-80% of the same year's cells. The survival and pupal mass of the potter wasp offspring were negatively associated with the presence and number of parasitized caterpillars inside the cells. We concluded that the collection of parasitized caterpillars by D. dimidiantipenne is frequent and costly. The persistence of this behavior may result from limited discrimination ability against parasitized prey by female potter wasps, or by their limited ability to exhibit choosiness under field conditions.

The combined effect of host and food availability on optimized parasitoid life-history traits based on a three-dimensional trade-off surface.

The reproductive success of many insects is considered to be limited by two main factors: the availability of mature eggs to lay (termed egg limitation) and the time to locate suitable hosts (termed time limitation). High host density in the environment is likely to enhance oviposition opportunities, thereby selecting for higher investment in egg supply. In contrast, a shortage of food (e.g. sugar sources) is likely to increase the risk of time limitation, thereby selecting for higher allocation to initial energy reserves. To our knowledge, the combined effect of host and food availability on these optimal life-history allocations has never been investigated. We thus modelled their simultaneous effects on a three-dimensional trade-off between initial investment in energy reserves, egg number and egg size, while focusing on insect parasitoids. The model was based on Monte Carlo simulations coupled with genetic algorithms, in order to identify the optimal life-history traits of a single simulated parasitoid female in an environment in which both hosts and food are present in varying densities. Our results reproduced the simple predictions described above. However, some novel predictions were also obtained, especially when specific interactions between the different factors were examined and their effects on the three-dimensional life-history surface were considered. The work sheds light on long-lasting debates regarding the relative importance of time versus egg limitation in determining insect life-history traits and highlights the complexity of life-history evolution, where several environmental factors act simultaneously on multiple traits.

What do we know about biological nitrogen fixation in insects? Evidence and implications for the insect and the ecosystem.

Many insects feed on a low-nitrogen diet, and the origin of their nitrogen supply is poorly understood. It has been hypothesized that some insects rely on nitrogen-fixing bacteria (diazotrophs) to supplement their diets. Nitrogen fixation by diazotrophs has been extensively studied and convincingly demonstrated in termites, while evidence for the occurrence and role of nitrogen fixation in the diet of other insects is less conclusive. Here, we summarize the methods to detect nitrogen fixation in insects and review the available evidence for its occurrence (focusing on insects other than termites). We distinguish between three aspects of nitrogen fixation investigations: (i) detecting the presence of potential diazotrophs; (ii) detecting the activity of the nitrogen-fixing enzyme; and (iii) detecting the assimilation of fixed nitrogen into the insect tissues. We show that although evidence from investigations of the first aspect reveals ample opportunities for interactions with potential diazotrophs in a variety of insects, demonstrations of actual biological nitrogen fixation and the assimilation of fixed nitrogen are restricted to very few insect groups, including wood-feeding beetles, fruit flies, leafcutter ants, and a wood wasp. We then discuss potential implications for the insect's fitness and for the ecosystem as a whole. We suggest that combining these multiple approaches is crucial for the study of nitrogen fixation in insects, and argue that further demonstrations are desperately needed in order to determine the relative importance of diazotrophs for insect diet and fitness, as well as to evaluate their overall impact on the ecosystem.

Gut bacteria of weevils developing on plant roots under extreme desert conditions.

BACKGROUND: Many phytophagous insects, whose diet is generally nitrogen-poor, rely on gut bacteria to compensate for nutritional deficits. Accordingly, we hypothesized that insects in desert environments may evolve associations with gut bacteria to adapt to the extremely low nutrient availability. For this, we conducted a systematic survey of bacterial communities in the guts of weevils developing inside mud chambers affixed to plant roots in the Negev Desert of Israel, based on 16S rRNA gene amplicon sequencing. RESULTS: Our analyses revealed that gut bacterial communities in weevil larvae were similar across a wide geographical range, but differed significantly from those of the mud chambers and of the surrounding soils. Nevertheless, a high proportion of bacteria (including all of the core bacteria) found in the weevils were also detected in the mud chambers and soils at low relative abundances. The genus Citrobacter (of the Enterobacteriaceae family) was the predominant group in the guts of all individual weevils. The relative abundance of Citrobacter significantly decreased at the pupal and adult stages, while bacterial diversity increased. A mini literature survey revealed that members of the genus Citrobacter are associated with nitrogen fixation, recycling of uric acid nitrogen, and cellulose degradation in different insects. CONCLUSIONS: The results suggest that although weevils could potentially acquire their gut bacteria from the soil, weevil host internal factors, rather than external environmental factors, were more important in shaping their gut bacterial communities, and suggest a major role for Citrobacter in weevil nutrition in this challenging environment. This study highlights the potential involvement of gut bacteria in the adaptation of insects to nutritional deficiencies under extreme desert conditions.

Wolbachia's role in mediating its flea's reproductive success differs according to flea origin.

Endosymbionts-microbes that live within and engage in prolonged and intimate associations with their hosts-are gaining recognition for their direct impact on plant and animal reproduction. Here we used the overlooked Wolbachia-flea system to explore the possibility that endosymbionts may also play a role as mediators in shaping the reproductive success of their hosts. We simultaneously quantified the Wolbachia density in field- and laboratory-originated fleas that fed and mated on rodents for either 5 or 10 days and assessed their body size and current reproductive success. By combining multigroup analysis and model selection approaches, we teased apart the contribution of the direct effects of the flea's physiological age and body size and the mediation effect of its Wolbachia endosymbionts on flea reproductive success, and we showed that the latter was stronger than the former. However, interestingly, the mediation effect was manifested only in laboratory-originated fleas, for which the increase in Wolbachia with age translated into lower reproductive success. These results suggest that some well-supported phenomena, such as aging effects, may be driven by endosymbionts and show once again that the role of endosymbionts in shaping the reproductive success of their host depends on their selective environment.

Lessons from the multitudes: insights from polyembryonic wasps for behavioral ecology.

Even for parasitic Hymenoptera, polyembryonic wasps are unusual creatures. Two features in particular, allow for novel exploration of major questions in behavioral ecology: the production of multiple offspring per egg and, in some species, the production of a soldier caste. Because final brood sizes of polyembryonic species are not constrained by trade-offs between current and future parental reproductive effort, we can clearly examine the selective forces at play that drive the balance between the number of offspring and their body size. Polyembryony also provides excellent opportunities to compare the performance of identical genotypes under different environmental conditions. Finally, polyembryonic species can provide unique tests of how genetic conflicts at multiple levels are resolved.

Tripartite symbiosis of plant-weevil-bacteria is a widespread phenomenon in the Negev Desert.

The weevil Conorhynchus palumbus develops in a mud chamber affixed to the roots of the summer annual plant Salsola inermis in the Negev Desert of Israel. The weevil carries nitrogen fixing bacteria, and evidence suggests that plants with weevils utilize the fixed nitrogen. To characterize the distribution, abundance and significance of this unique interaction, we surveyed Salsola plants in 16 sites throughout the Negev Desert. We excavated ~100 plants from each site, recorded the presence of weevils in their roots, and characterized the soil properties in each site. Weevil mud chambers were present in all of the sampled sites and their abundance was positively correlated with soil nitrogen content and with plant size, and negatively correlated with soil grain-size. Intriguingly, we found two additional weevil species-Menecleonus virgatus and Maximus mimosae-residing in mud chambers on Salsola roots, and found one additional Salsola species-S. incanescens-accommodating weevils. Nitrogen fixing bacteria were found in weevil larvae of the two additional species and at multiple sites. Overall, our findings suggest that potentially beneficial associations between weevils and plants may be more common than previously acknowledged, and may play an important role in this desert ecosystem.

Factors shaping life history traits of two proovigenic parasitoids.

What shapes the relative investment in reproduction versus survival of organisms is among the key questions in life history. Proovigenic insects mature all their eggs prior to emergence and are short lived, providing a unique opportunity to quantify their lifetime investments in the different functions. We investigated the initial eggloads and longevity of 2 proovigenic parasitoid wasps: Anagrus erythroneurae and Anagrus daanei, (Hymenoptera: Mymaridae) that develop within leafhopper eggs in both agricultural vineyards and natural riparian habitats in Northern California. We collected Vitis spp. leaves containing developing parasitoids from 3 natural sites (Knight Landing, American River and Putah Creek) and 3 agricultural vineyards (Solano Farm, Davis Campus and Village Homes). We recorded eggloads at parasitoid emergence and female parasitoid longevity with or without honey-feeding. Theory predicts that parasitoids from vineyards (where hosts are abundant) would have higher initial eggloads and lower longevity compared with parasitoids from riparian habitats (where hosts are scarce). Although host density and parasitoid eggloads were, indeed, higher in vineyards than in riparian habitats, parasitoid longevity did not follow the predicted pattern. Longevity without feeding differed among field sites, but it was not affected by habitat type (natural vs agricultural), whereas longevity with feeding was not significantly affected by any of the examined factors. Moreover, longevity was positively, rather than negatively, correlated with eggloads at the individual level, even after correcting for parasitoid body size. The combined results suggest a more complex allocation mechanism than initially predicted, and the possibility of variation in host quality that is independent of size.

Plant-mediated resource partitioning by coexisting parasitoids.

Although it has been frequently suggested that resource partitioning of species coexisting at the same trophic level can be mediated by interactions with species at non-adjacent trophic levels, empirical evidence supporting this claim is scarce. Here we demonstrate that plants may mediate resource partitioning for two parasitoids that share the same herbivorous host. The tephritid fly Tephritis femoralis is the primary pre-dispersal seed predator of two Asteraceae species, Saussurea nigrescens and Anaphalis flavescens, both of which dominate the plant community in the alpine meadows of the Tibetan Plateau. Field surveys and molecular barcoding analyses showed that the identity of the fly's main predator depended on the plant in which the fly developed. Tephritid flies that developed in S. nigrescens were preyed upon mainly by the parasitoid wasp Pteromalus albipennis, while the parasitoid Mesopolobus sp. was the main predator of flies that developed in A. flavescens. Microcosm experiments revealed that P. albipennis could not exploit the host flies within the capitula of A. flavescens due to food limitation (capitula are too small), while Mesopolobus sp. could not exploit the host flies within the capitula of S. nigrescens due to its inability to reach the host with its ovipositor (capitula are too large). Such bottom-up control of plant species traits may facilitate the coexistence of parasitoid wasps sharing a common host in this system. We suggest that interactions between non-adjacent trophic levels may potentially promote species coexistence and diversity in biological communities.

The effect of virus-blocking Wolbachia on male competitiveness of the dengue vector mosquito, Aedes aegypti.

BACKGROUND: The bacterial endosymbiont Wolbachia blocks the transmission of dengue virus by its vector mosquito Aedes aegypti, and is currently being evaluated for control of dengue outbreaks. Wolbachia induces cytoplasmic incompatibility (CI) that results in the developmental failure of offspring in the cross between Wolbachia-infected males and uninfected females. This increases the relative success of infected females in the population, thereby enhancing the spread of the beneficial bacterium. However, Wolbachia spread via CI will only be feasible if infected males are sufficiently competitive in obtaining a mate under field conditions. We tested the effect of Wolbachia on the competitiveness of A. aegypti males under semi-field conditions. METHODOLOGY/PRINCIPAL FINDINGS: In a series of experiments we exposed uninfected females to Wolbachia-infected and uninfected males simultaneously. We scored the competitiveness of infected males according to the proportion of females producing non-viable eggs due to incompatibility. We found that infected males were equally successful to uninfected males in securing a mate within experimental tents and semi-field cages. This was true for males infected by the benign wMel Wolbachia strain, but also for males infected by the virulent wMelPop (popcorn) strain. By manipulating male size we found that larger males had a higher success than smaller underfed males in the semi-field cages, regardless of their infection status. CONCLUSIONS/SIGNIFICANCE: The results indicate that Wolbachia infection does not reduce the competitiveness of A. aegypti males. Moreover, the body size effect suggests a potential advantage for lab-reared Wolbachia-males during a field release episode, due to their better nutrition and larger size. This may promote Wolbachia spread via CI in wild mosquito populations and underscores its potential use for disease control.