Facultatively ectoparasitic mites as vectors for entomopathogenic bacteria in Drosophila.

Opportunistic bacterial infections are common in insect populations but there is little information on how they are acquired or transmitted. We tested the hypothesis that Macrocheles mites can transmit systemic bacterial infections between Drosophila hosts. We found that 24% of mites acquired detectable levels of bacteria after feeding on infected flies and 87% of infected mites passed bacteria to naïve recipient flies. The probability that a mite could pass Serratia from an infected donor fly to a naïve recipient fly was 27.1%. These data demonstrate that Macrocheles mites are capable of serving as vectors of bacterial infection between insects.

The effects of habitat type and pathogen infection on tick host-seeking behaviour.

Tick-borne pathogens pose a significant risk to livestock, wildlife and public health. Host-seeking behaviours may depend on a combination of infection status and environmental factors. Here, we assessed the effects of habitat type and pathogen infection on host-seeking behaviour (questing) in the lone star tick, Amblyomma americanum. Ticks were collected using a tick drag from two different habitat types: xeric hammock and successional hardwood forests. Using a standardized assay, we recorded the likelihood of questing for each tick, the average height quested and total time spent questing and then tested each tick for the presence of Rickettsia spp. and Ehrlichia spp. using conventional polymerase chain reaction. We did not detect Ehrlichia in any ticks, although 30% tested positive for Rickettsia amblyommatis, a member of the Rickettsia spotted fever group. Ticks infected with R. amblyommatis spent less time questing compared to uninfected ticks, with infected ticks spending 85 s on average questing and uninfected ticks spending 112 s. Additionally, ticks collected from xeric hammock habitats spent over twice as long questing compared to ticks from successional hardwood forests. Ticks from xeric hammock spent 151 s on average questing while ticks from successional hardwood forest spent only 58 s during a 10-min observation period. These results demonstrate that habitat type and infection status can influence tick host-seeking behaviours, which can play a pivotal role in disease dynamics.

Prevalence and intensity of avian malaria in a quail hybrid zone.

Hybrid zones have been described as natural laboratories by researchers who study speciation and the various mechanisms that may affect gene flow. The evolutionary consequences of hybridization depend not only on reproductive compatibility between sympatric species, but also on factors like vulnerability to each other's predators and parasites. We examined infection patterns of the blood parasite Haemoproteus lophortyx, a causative agent of avian malaria, at a site in the contact zone between California quail (Callipepla californica) and Gambel's quail (C. gambelii). Controlling for the potential influence of sex and year, we tested whether species identity predicted infection status and intensity. We found that infection prevalence was lower in California and hybrid quail compared with Gambel's quail. However, infected California and hybrid quail had higher infection intensities than Gambel's quail. California and hybrid quail exhibited no significant differences in prevalence or intensity of infection. These findings suggest that infection by H. lophortyx has the potential to influence species barrier dynamics in this system; however, more work is necessary to determine the exact evolutionary consequences of this blood parasite on hybridization.

Spatial proximity and prey vibratory cues influence collective hunting in social spiders.

Social spiders are thought to predominantly receive information about their environment through vibrational cues. Thus, group living introduces the challenge of distinguishing useful vibrational information from the background noise of nestmates. Here we investigate whether spatial proximity between colony-mates may allow social spiders (Stegodyphus dumicola) to reduce background noise that might obstruct vibrational information from prey. To do so, we constructed experimental colonies and measured whether the number of spiders in proximity to one another whilst resting could predict the number of spiders that participated in prey capture. Additionally, we exposed spider colonies to five different simulated vibrational cues mimicking prey to determine which cue types spiders were most responsive to. We found that the number of spiders huddled together prior to foraging trials was positively correlated with the number of spiders participating in collective foraging. Furthermore, colonies responded more quickly to pulsed vibrational cues over other types of vibrational patterns. Together these data reveal that both social interactions and prey cues shape how social sit-and-wait predators experience and respond to their environment.

Tadpole body size and behaviour alter the social acquisition of a defensive bacterial symbiont.

Individual differences in host phenotypes can generate heterogeneity in the acquisition and transmission of microbes. Although this has become a prominent factor of disease epidemiology, host phenotypic variation might similarly underlie the transmission of microbial symbionts that defend against pathogen infection. Here, we test whether host body size and behaviour influence the social acquisition of a skin bacterium, Janthinobacterium lividum, which in some hosts can confer protection against infection by Batrachochytrium dendrobatidis, the causative agent of the amphibian skin disease chytridiomycosis. We measured body size and boldness (time spent in an open field) of green frog tadpoles and haphazardly constructed groups of six individuals. In some groups, we exposed one individual in each group to J. lividum and, in other groups, we inoculated a patch of aquarium pebbles to J. lividum. After 24 h, we swabbed each individual to estimate the presence of J. lividum on their skin. On average, tadpoles acquired nearly four times more bacteria when housed with an exposed individual compared to those housed with a patch of inoculated substrate. When tadpoles were housed with an exposed group-mate, larger and 'bolder' individuals acquired more bacteria. These data suggest that phenotypically biased acquisition of defensive symbionts might generate biased patterns of mortality from the pathogens against which they protect.

Social spider webs harbour largely consistent bacterial communities across broad spatial scales.

Social animals that live in domiciles constructed from biomaterials may facilitate microbial growth. Spider webs are one of the most conspicuous biomaterials in nature, yet almost nothing is known about the potential for webs to harbour microbes, even in social spiders that live in dense, long-term aggregations. Here, we tested whether the dominant bacteria present in social spider webs vary across sampling localities and whether the more permanent retreat web harbours compositionally distinct microbes from the more ephemeral capture webs in the desert social spider, Stegodyphus dumicola. We also sampled spider cuticles and prey items in a subset of colonies. We found that spider colonies across large spatial scales harboured similar web-associated bacterial communities. We also found substantial overlap in bacterial community composition between spider cuticle, prey and web samples. These data suggest that social spider webs can harbour characteristic microbial communities and potentially facilitate microbial transmission among individuals, and this study serves as the first step towards understanding the microbial ecology of these peculiar animal societies.

The emerging field of venom-microbiomics for exploring venom as a microenvironment, and the corresponding Initiative for Venom Associated Microbes and Parasites (iVAMP).

Venom is a known source of novel antimicrobial natural products. The substantial, increasing number of these discoveries have unintentionally culminated in the misconception that venom and venom-producing glands are largely sterile environments. Culture-dependent and -independent studies on the microbial communities in venom microenvironments reveal the presence of archaea, algae, bacteria, fungi, protozoa, and viruses. Venom-centric microbiome studies are relatively sparse to date with the adaptive advantages that venom-associated microbes might offer to their hosts, or that hosts might provide to venom-associated microbes, remaining largely unknown. We highlight the potential for the discovery of venom microbiomes within the adaptive landscape of venom systems. The considerable number of convergently evolved venomous animals, juxtaposed with the comparatively few known studies to identify microbial communities in venom, provides new possibilities for both biodiversity and therapeutic discoveries. We present an evidence-based argument for integrating microbiology as part of venomics (i.e., venom-microbiomics) and introduce iVAMP, the Initiative for Venom Associated Microbes and Parasites (https://ivamp-consortium.github.io/), as a growing collaborative consortium. We express commitment to the diversity, inclusion and scientific collaboration among researchers interested in this emerging subdiscipline through expansion of the iVAMP consortium.

Exposure to cuticular bacteria can alter host behavior in a funnel-weaving spider.

Contact with environmental microbes are arguably the most common species interaction in which any animal participates. Studies have noted diverse relationships between hosts and resident microbes, which can have strong consequences for host development, physiology, and behavior. Many of these studies focus specifically on pathogens or beneficial microbes, while the benign microbes, of which the majority of bacteria could be described, are often ignored. Here, we explore the nature of the relationships between the grass spider Agelenopsis pennsylvanica and bacteria collected from their cuticles in situ. First, using culture-based methods, we identified a portion of the cuticular bacterial communities that are naturally associated with these spiders. Then, we topically exposed spiders to a subset of these bacterial monocultures to estimate how bacterial exposure may alter 3 host behavioral traits: boldness, aggressiveness, and activity level. We conducted these behavioral assays 3 times before and 3 times after topical application, and compared the changes observed in each trait with spiders that were exposed to a sterile control treatment. We identified 9 species of bacteria from the cuticles of 36 spiders and exposed groups of 20 spiders to 1 of 4 species of cuticular bacteria. We found that exposure to Dermacoccus nishinomiyaensis and Staphylococcus saprophyticus was associated with a 10-fold decrease in the foraging aggressiveness of spiders toward prey in their web. Since bacterial exposure did not have survival consequences for hosts, these data suggest that interactions with cuticular bacteria, even non-pathogenic bacteria, could alter host behavior.

Erratum: Exposure to cuticular bacteria can alter host behavior in a funnel-weaving spider.

[This corrects the article DOI: 10.1093/cz/zox064.]

The primary case is not enough: Variation among individuals, groups and social networks modify bacterial transmission dynamics.

The traits of the primary case of an infectious disease outbreak, and the circumstances for their aetiology, potentially influence the trajectory of transmission dynamics. However, these dynamics likely also depend on the traits of the individuals with whom the primary case interacts. We used the social spider Stegodyphus dumicola to test how the traits of the primary case, group phenotypic composition and group size interact to facilitate the transmission of a GFP-labelled cuticular bacterium. We also compared bacterial transmission across experimentally generated "daisy-chain" vs. "star" networks of social interactions. Finally, we compared social network structure across groups of different sizes. Groups of 10 spiders experienced more bacterial transmission events compared to groups of 30 spiders, regardless of groups' behavioural composition. Groups containing only one bold spider experienced the lowest levels of bacterial transmission regardless of group size. We found no evidence for the traits of the primary case influencing any transmission dynamics. In a second experiment, bacteria were transmitted to more individuals in experimentally induced star networks than in daisy-chains, on which transmission never exceeded three steps. In both experimental network types, transmission success depended jointly on the behavioural traits of the interacting individuals; however, the behavioural traits of the primary case were only important for transmission on star networks. Larger social groups exhibited lower interaction density (i.e. had a low ratio of observed to possible connections) and were more modular, i.e. they had more connections between nodes within a subgroup and fewer connections across subgroups. Thus, larger groups may restrict transmission by forming fewer interactions and by isolating subgroups that interacted with the primary case. These findings suggest that accounting for the traits of single exposed hosts has less power in predicting transmission dynamics compared to the larger scale factors of the social groups in which they reside. Factors like group size and phenotypic composition appear to alter social interaction patterns, which leads to differential transmission of microbes.

Queen presence mediates the relationship between collective behaviour and disease susceptibility in ant colonies.

The success of social living can be explained, in part, by a group's ability to execute collective behaviours unachievable by solitary individuals. However, groups vary in their ability to execute these complex behaviours, often because they vary in their phenotypic composition. Group membership changes over time due to mortality or emigration, potentially leaving groups vulnerable to ecological challenges in times of flux. In some societies, the loss of important individuals (e.g. leaders, elites and queens) may have an especially detrimental effect on groups' ability to deal with these challenges. Here, we test whether the removal of queens in colonies of the acorn ant Temnothorax curvispinosus alters their ability to execute important collective behaviours and survive outbreaks of a generalist entomopathogen. We employed a split-colony design where one half of a colony was maintained with its queen, while the other half was separated from the queen. We then tested these subcolonies' performance in a series of collective behaviour assays and finally exposed colonies to the entomopathogenic fungus Metarhizium robertsii by exposing two individuals from the colony and then sealing them back into the nest. We found that queenright subcolonies outperformed their queenless counterparts in nearly all collective behaviours. Queenless subcolonies were also more vulnerable to mortality from disease. However, queenless groups that displayed more interactions with brood experienced greater survivorship, a trend not present in queenright subcolonies. Queenless subcolonies that engage in more brood interactions may have had more resources available to cope with two physiological challenges (ovarian development after queen loss and immune activation after pathogen exposure). Our results indicate that queen presence can play an integral role in colony behaviour, survivorship and their relationship. They also suggest that interactions between workers and brood are integral to colonies survival. Overall, a social group's history of social reorganization may have strong consequences on their collective behaviours and their vulnerability to disease outbreaks.

The Effect of Keystone Individuals on Collective Outcomes Can Be Mediated through Interactions or Behavioral Persistence.

Collective behavior emerges from interactions among group members who often vary in their behavior. The presence of just one or a few keystone individuals, such as leaders or tutors, may have a large effect on collective outcomes. These individuals can catalyze behavioral changes in other group members, thus altering group composition and collective behavior. The influence of keystone individuals on group function may lead to trade-offs between ecological situations, because the behavioral composition they facilitate may be suitable in one situation but not another. We use computer simulations to examine various mechanisms that allow keystone individuals to exert their influence on group members. We further discuss a trade-off between two potentially conflicting collective outcomes, cooperative prey attack and disease dynamics. Our simulations match empirical data from a social spider system and produce testable predictions for the causes and consequences of the influence of keystone individuals on group composition and collective outcomes. We find that a group's behavioral composition can be impacted by the keystone individual through changes to interaction patterns or behavioral persistence over time. Group behavioral composition and the mechanisms that drive the distribution of phenotypes influence collective outcomes and lead to trade-offs between disease dynamics and cooperative prey attack.

Personality composition alters the transmission of cuticular bacteria in social groups.

The initial stages of a disease outbreak can determine the magnitude of the ensuing epidemic. Though rarely tested in unison, two factors with important consequences for the transmission dynamics of infectious agents are the collective traits of the susceptible population and the individual traits of the index case (i.e. 'patient zero'). Here, we test whether the personality composition of a social group can explain horizontal transmission dynamics of cuticular bacteria using the social spider Stegodyphus dumicola We exposed focal spiders of known behavioural phenotypes with a GFP-transformed cuticular bacterium (Pantoea sp.) and placed them in groups of 10 susceptible individuals (i.e. those with no experience with this bacterium). We measured bacterial transmission to groups composed of either all shy spiders, 10% bold spiders or 40% bold spiders. We found that colonies with 40% bold spiders experienced over twice the incidence of transmission compared to colonies with just 10% bold individuals after only 24 h of interaction. Colonies of all shy spiders experienced an intermediate degree of transmission. Interestingly, we did not detect an effect of the traits of the index case on transmission. These data suggest that the phenotypic composition of the susceptible population can have a greater influence on the degree of early transmission events than the traits of the index case.

Individual differences in boldness influence patterns of social interactions and the transmission of cuticular bacteria among group-mates.

Despite the importance of host attributes for the likelihood of associated microbial transmission, individual variation is seldom considered in studies of wildlife disease. Here, we test the influence of host phenotypes on social network structure and the likelihood of cuticular bacterial transmission from exposed individuals to susceptible group-mates using female social spiders (Stegodyphus dumicola). Based on the interactions of resting individuals of known behavioural types, we assessed whether individuals assorted according to their behavioural traits. We found that individuals preferentially interacted with individuals of unlike behavioural phenotypes. We next applied a green fluorescent protein-transformed cuticular bacterium,Pantoeasp., to individuals and allowed them to interact with an unexposed colony-mate for 24 h. We found evidence for transmission of bacteria in 55% of cases. The likelihood of transmission was influenced jointly by the behavioural phenotypes of both the exposed and susceptible individuals: transmission was more likely when exposed spiders exhibited higher 'boldness' relative to their colony-mate, and when unexposed individuals were in better body condition. Indirect transmission via shared silk took place in only 15% of cases. Thus, bodily contact appears key to transmission in this system. These data represent a fundamental step towards understanding how individual traits influence larger-scale social and epidemiological dynamics.

The Achilles' heel hypothesis: misinformed keystone individuals impair collective learning and reduce group success.

Many animal societies rely on highly influential keystone individuals for proper functioning. When information quality is important for group success, such keystone individuals have the potential to diminish group performance if they possess inaccurate information. Here, we test whether information quality (accurate or inaccurate) influences collective outcomes when keystone individuals are the first to acquire it. We trained keystone or generic individuals to attack or avoid novel stimuli and implanted these trained individuals within groups of naive colony-mates. We subsequently tracked how quickly groups learned about their environment in situations that matched (accurate information) or mismatched (inaccurate information) the training of the trained individual. We found that colonies with just one accurately informed individual were quicker to learn to attack a novel prey stimulus than colonies with no informed individuals. However, this effect was no more pronounced when the informed individual was a keystone individual. In contrast, keystones with inaccurate information had larger effects than generic individuals with identical information: groups containing keystones with inaccurate information took longer to learn to attack/avoid prey/predator stimuli and gained less weight than groups harbouring generic individuals with identical information. Our results convey that misinformed keystone individuals can become points of vulnerability for their societies.

Cuticular bacteria appear detrimental to social spiders in mixed but not monoculture exposure.

Much of an animal's health status, life history, and behavior are dictated by interactions with its endogenous and exogenous bacterial communities. Unfortunately, interactions between hosts and members of their resident bacterial community are often ignored in animal behavior and behavioral ecology. Here, we aim to identify the nature of host-microbe interactions in a nonmodel organism, the African social spider Stegodyphus dumicola. We collected and identified bacteria from the cuticles of spiders in situ and then exposed spiders to bacterial monocultures cultures via topical application or injection. We also topically inoculated spiders with a concomitant "cocktail" of bacteria and measured the behavior of spiders daily for 24 days after inoculation. Lastly, we collected and identified bacteria from the cuticles of prey items in the capture webs of spiders, and then fed spiders domestic crickets which had been injected with these bacteria. We also injected 1 species of prey-borne bacteria into the hemolymph of spiders. Only Bacillus thuringiensis caused increased mortality when injected into the hemolymph of spiders, whereas no bacterial monocultures caused increased mortality when applied topically, relative to control solutions. However, a bacterial cocktail of cuticular bacteria caused weight loss and mortality when applied topically, yet did not detectibly alter spider behavior. Consuming prey injected with prey-borne bacteria was associated with an elongated lifespan in spiders. Thus, indirect evidence from multiple experiments suggests that the effects of these bacteria on spider survivorship appear contingent on their mode of colonization and whether they are applied in monoculture or within a mixed cocktail. We urge that follow-up studies should test these host-microbe interactions across different social contexts to determine the role that microbes play in colony performance.

Warring arthropod societies: Social spider colonies can delay annihilation by predatory ants via reduced apparency and increased group size.

Sociality provides individuals with benefits via collective foraging and anti-predator defense. One of the costs of living in large groups, however, is increased apparency to natural enemies. Here, we test how the individual-level and collective traits of spider societies can increase the risk of discovery and death by predatory ants. We transplanted colonies of the social spider Stegodyphus dumicola into a habitat dense with one of their top predators, the pugnacious ant Anoplolepis custodiens. With three different experiments, we test how colony-wide survivorship in a predator-dense habitat can be altered by colony apparency (i.e., the presence of a capture web), group size, and group composition (i.e., the proportion of bold and shy personality types present). We also test how spiders' social context (i.e., living solitarily vs. among conspecifics) modifies their behaviour toward ants in their capture web. Colonies with capture webs intact were discovered by predatory ants on average 25% faster than colonies with the capture web removed, and all discovered colonies eventually collapsed and succumbed to predation. However, the lag time from discovery by ants to colony collapse was greater for colonies containing more individuals. The composition of individual personality types in the group had no influence on survivorship. Spiders in a social group were more likely to approach ants caught in their web than were isolated spiders. Isolated spiders were more likely to attack a safe prey item (a moth) than they were to attack ants and were more likely to retreat from ants after contact than they were after contact with moths. Together, our data suggest that the physical structures produced by large animal societies can increase their apparency to natural enemies, though larger groups can facilitate a longer lag time between discovery and demise. Lastly, the interaction between spiders and predatory ants seems to depend on the social context in which spiders reside.

Integrating animal personality into insect population and community ecology.

Despite the recent surge of interest in the concept of animal personalities, that is, temporally consistent individual differences in behavior, few studies have integrated intraspecific behavioral variation in population or community ecology. Insects and other arthropods provide ideal model systems to study how intraspecific behavioral variation affects phenomena in ecology. This is due to the fact that arthropods not only are highly amenable to experimental manipulation, but they also allow us to answer general ecological questions on multiple scales of biological organization. Herein, we review recent developments and views on how the framework of animal personality could provide a deeper understanding of classic issues in (1) population ecology (e.g., local adaptation, dispersal, and invasion), (2) community ecology (e.g., food webs and ecosystem engineering), and (3) more insect-focused topics such as metamorphosis and pollination biology.