Chronic stress and captivity alter the cloacal microbiome of a wild songbird.

There are complex interactions between an organism's microbiome and its response to stressors, often referred to as the 'gut-brain axis'; however, the ecological relevance of this axis in wild animals remains poorly understood. Here, we used a chronic mild stress protocol to induce stress in wild-caught house sparrows (Passer domesticus), and compared microbial communities among stressed animals, those recovering from stress, captive controls (unstressed) and a group not brought into captivity. We assessed changes in microbial communities and abundance of shed microbes by culturing cloacal samples on multiple media to select for aerobic and anaerobic bacteria and fungi. We complemented this with cultivation-independent 16S and ITS rRNA gene amplification and sequencing, pairing these results with host physiological and immune metrics, including body mass change, relative spleen mass and plasma corticosterone concentrations. We found significant effects of stress and captivity on the house sparrow microbiomes, with stress leading to an increased relative abundance of endotoxin-producing bacteria - a possible mechanism for the hyperinflammatory response observed in captive avians. While we found evidence that the microbiome community partially recovers after stress cessation, animals may lose key taxa, and the abundance of endotoxin-producing bacteria persists. Our results suggest an overall link between chronic stress, host immune system and the microbiome, with the loss of potentially beneficial taxa (e.g. lactic acid bacteria), and an increase in endotoxin-producing bacteria due to stress and captivity. Ultimately, consideration of the host's microbiome may be useful when evaluating the impact of stressors on individual and population health.

Growsafe: A Chemical Method To Deactivate Cultivated Microorganisms Using Low-Cost Kitchen Supplies.

One limitation to engaging K-12 students and the public with microorganisms is the inability to cultivate and dispose of bacterial and fungal samples safely without expensive equipment or services. This barrier has been amplified with remote learning modalities and laboratory closures driven by safety precautions due to the COVID-19 pandemic. At-home lab kits are being used to bring hands-on experience in microorganism cultivation to students learning remotely, but these kits often fail to take into full consideration the safety aspects or the costs associated with microorganism disposal, limiting which experiments can be performed at home. Here, we outline a method that makes cultivating and deactivating microorganisms accessible to the public through low-cost and readily available equipment. This method reduces exposure to microorganisms by forgoing the need to open petri plates for chemical deactivation with sanitizing reagents. This technique may benefit remote K-12 and postsecondary students, students wishing to get hands-on microbiology research experience, and members of the public interested in cultivating microorganisms to contribute to citizen science efforts or for creative art applications.

Does HIV infection increase male sexual behavior?

After 40 years of intense study on HIV/AIDS, scientists have identified, among other things, at risk populations, stages of disease progression and treatment strategies. What has received less attention is the possibility that infection might elicit an increase in sexual behavior in humans. In 2000, Starks and colleagues speculated that HIV infection could alter host behavior in a manner that facilitated the spread of the virus. Retrospective and self-report data from five studies now support this hypothesis. Individuals with acute-versus nonacute-stage infections report more sexual partners and more frequent risky sex. Additionally, male sexual behavior increases nonlinearly with HIV viral load, and data suggest a potential threshold viral level above which individuals are more likely to engage in risky sexual behavior. Taken together, these data suggest that HIV infection influences male sexual behavior in a manner beneficial to the virus. Here, we present these findings, highlight their limitations and discuss alternative perspectives. We argue for increased testing of this hypothesis and advocate for increased public health measures to mitigate the putative impact on male sexual behavior. Lay Summary In 2000, Starks and colleagues speculated that HIV infection could alter host behavior in a manner that facilitated the spread of the virus. Retrospective and self-report data from five studies now support this hypothesis. We argue for increased testing of this hypothesis and advocate for increased public health measures to mitigate the putative impact on male sexual behavior.

An evolutionary perspective on night terrors.

Night terrors, also known as sleep terrors, are an early childhood parasomnia characterized by screams or cries, behavioral manifestations of extreme fear, difficulty waking and inconsolability upon awakening. The mechanism causing night terrors is unknown, and a consistently successful treatment has yet to be documented. Here, we argue that cultural practices have moved us away from an ultimate solution: cosleeping. Cosleeping is the norm for closely related primates and for humans in non-Western cultures. In recent years, however, cosleeping has been discouraged by the Western medical community. From an evolutionary perspective, cosleeping provides health and safety benefits for developing children. We discuss night terrors, and immediate and long-term health features, with respect to cosleeping, room-sharing and solitary sleeping. We suggest that cosleeping with children (≥1-year-old) may prevent night terrors and that, under certain circumstances, cosleeping with infants (≤11-months-old) is preferable to room-sharing, and both are preferable to solitary sleeping.

Seasonality of honey bee (Apis mellifera) micronutrient supplementation and environmental limitation.

Honey bees (Apis mellifera) obtain micronutrients from floral resources and "dirty", or turbid, water. Past research suggests that honey bees drink dirty water to supplement the micronutrients in their floral diet, however, there is no research that directly investigates how floral micronutrient content varies with water preferences, or how micronutrients in honey bees themselves vary seasonally. In this study, we used chemical analyses (ICP-OES) to investigate seasonal variation of micronutrients in honey bee workers and floral resources in the field. We found that honey bees likely use mineralized water to supplement their floral diet and may be limited by availability of calcium and potassium. Our results also suggest that honey bees may seasonally seek specific micronutrients, perhaps in preparation for overwintering.

The European wool-carder bee (Anthidium manicatum) eavesdrops on plant volatile organic compounds (VOCs) during trichome collection.

The plant-pollinator relationship is generally considered mutualistic. This relationship is less clear, however, when pollinators also cause tissue damage. Some Megachilidae bees collect plant material for nests from the plants they pollinate. In this study, we examined the relationship between Anthidium manicatum, the European wool-carder bee, and the source of its preferred nesting material - Stachys byzantina, lamb's ear. Female A. manicatum use their mandibles to trim trichomes from plants for nesting material (a behaviour dubbed "carding"). Using volatile organic compound (VOC) headspace analysis and behavioural observations, we explored (a) how carding effects S. byzantina and (b) how A. manicatum may choose specific S. byzantina plants. We found that removal of trichomes leads to a dissimilar VOC bouquet compared to intact leaves, with a significant increase in VOC detection following damage. A. manicatum also visit S. byzantina plants with trichomes removed at a greater frequency compared to plants with trichomes intact. Our data suggest that A. manicatum eavesdrop on VOCs produced by damaged plants, leading to more carding damage for individual plants due to increased detectability by A. manicatum. Accordingly, visitation by A. manicatum to S. byzantina may incur both a benefit (pollination) and cost (tissue damage) to the plant.

The emerging contribution of social wasps to grape rot disease ecology.

Grape sour (bunch) rot is a polymicrobial disease of vineyards that causes millions of dollars in lost revenue per year due to decreased quality of grapes and resultant wine. The disease is associated with damaged berries infected with a community of acetic acid bacteria, yeasts, and filamentous fungi that results in rotting berries with high amounts of undesirable volatile acidity. Many insect species cause the initial grape berry damage that can lead to this disease, but most studies have focused on the role of fruit flies in facilitating symptoms and vectoring the microorganisms of this disease complex. Like fruit flies, social wasps are abundant in vineyards where they feed on ripe berries and cause significant damage, while also dispersing yeasts involved in wine fermentation. Despite this, their possible role in disease facilitation and dispersal of grape rots has not been explored. We tested the hypothesis that the paper wasp Polistes dominulus could facilitate grape sour rot in the absence of other insect vectors. Using marker gene sequencing we characterized the bacterial and fungal community of wild-caught adults. We used a sterilized foraging arena to determine if these wasps transfer viable microorganisms when foraging. We then tested if wasps harboring their native microbial community, or those inoculated with sour rot, had an effect on grape sour rot incidence and severity using a laboratory foraging arena. We found that all wasps harbor some portion of the sour rot microbial community and that they have the ability to transfer viable microorganisms when foraging. Foraging by inoculated and uninoculated wasps led to an increase in berry rot disease symptom severity and incidence. Our results indicate that paper wasps can facilitate sour rot diseases in the absence of other vectors and that the mechanism of this facilitation may include both increasing host susceptibility and transmitting these microbial communities to the grapes. Social wasps are understudied but relevant players in the sour rot ecology of vineyards.

Vasculature of the hive: heat dissipation in the honey bee (Apis mellifera) hive.

Eusocial insects are distinguished by their elaborate cooperative behavior and are sometimes defined as superorganisms. As a nest-bound superorganism, individuals work together to maintain favorable nest conditions. Residing in temperate environments, honey bees (Apis mellifera) work especially hard to maintain brood comb temperature between 32 and 36 °C. Heat shielding is a social homeostatic mechanism employed to combat local heat stress. Workers press the ventral side of their bodies against heated surfaces, absorb heat, and thus protect developing brood. While the absorption of heat has been characterized, the dissipation of absorbed heat has not. Our study characterized both how effectively worker bees absorb heat during heat shielding, and where worker bees dissipate absorbed heat. Hives were experimentally heated for 15 min during which internal temperatures and heat shielder counts were taken. Once the heat source was removed, hives were photographed with a thermal imaging camera for 15 min. Thermal images allowed for spatial tracking of heat flow as cooling occurred. Data indicate that honey bee workers collectively minimize heat gain during heating and accelerate heat loss during cooling. Thermal images show that heated areas temporarily increase in size in all directions and then rapidly decrease to safe levels (<37 °C). As such, heat shielding is reminiscent of bioheat removal via the cardiovascular system of mammals.

Actinomycetes with antimicrobial activity isolated from paper wasp (Hymenoptera: Vespidae: Polistinae) nests.

Actinomycetes-a group of antimicrobial producing bacteria-have been successfully cultured and characterized from the nest material of diverse arthropods. Some are symbionts that produce antimicrobial chemicals found to protect nest brood and resources from pathogenic microbes. Others have no known fitness relationship with their associated insects, but have been found to produce antimicrobials in vitro. Consequently, insect nest material is being investigated as a new source of novel antimicrobial producing actinomycetes, which could be harnessed for therapeutic potential. To extend studies of actinomycete-insect associations beyond soil-substrate dwelling insects and wood boring excavators, we conducted a preliminary assessment of the actinomycetes within the nests of the paper wasp, Polistes dominulus (Christ). We found that actinomycetes were readily cultured from nest material across multiple invasive P. dominulus populations-including members of the genera Streptomyces, Micromonospora, and Actinoplanes. Thirty of these isolates were assayed for antimicrobial activity against the challenge bacteria Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, Serratia marcescens, and Bacillus subtilis. Sixty percent of isolates inhibited the growth of at least one challenge strain. This study provides the first assessment of bacteria associated with nests of P. dominulus, and the first record of antimicrobial producing actinomycetes isolated from social wasps. We provide a new system to explore nest associated actinomycetes from a ubiquitous and cosmopolitan group of insects.

Within- and across-colony effects of hyperpolyandry on immune function and body condition in honey bees (Apis mellifera).

Honey bees (Apis mellifera) have become a model system for studies on the influence of genetic diversity on disease. Honey bee queens mate with a remarkably high number of males-up to 29 in the current study-from which they produce a colony of genetically diverse daughter workers. Recent evidence suggests a significant benefit of intracolony genetic diversity on disease resistance. Here, we explored the relationship between the level of genetic diversity and multiple physiological mechanisms of cellular and humoral immune defense (encapsulation response and phenoloxidase activity). We also investigated an effect of genetic diversity on a measure of body condition (fat body mass). While we predicted that mean colony phenoloxidase activity, encapsulation response, and fat body mass would show a positive relationship with increased intracolonial genetic diversity, we found no significant relationship between genetic diversity and these immune measures, and found no consistent effect on body condition. These results suggest that high genetic diversity as a result of extreme polyandry may have little bearing on the physiological mechanisms of immune function at naturally occurring mating levels in honey bees.

Evolution of a major drug metabolizing enzyme defect in the domestic cat and other felidae: phylogenetic timing and the role of hypercarnivory.

The domestic cat (Felis catus) shows remarkable sensitivity to the adverse effects of phenolic drugs, including acetaminophen and aspirin, as well as structurally-related toxicants found in the diet and environment. This idiosyncrasy results from pseudogenization of the gene encoding UDP-glucuronosyltransferase (UGT) 1A6, the major species-conserved phenol detoxification enzyme. Here, we established the phylogenetic timing of disruptive UGT1A6 mutations and explored the hypothesis that gene inactivation in cats was enabled by minimal exposure to plant-derived toxicants. Fixation of the UGT1A6 pseudogene was estimated to have occurred between 35 and 11 million years ago with all extant Felidae having dysfunctional UGT1A6. Out of 22 additional taxa sampled, representative of most Carnivora families, only brown hyena (Parahyaena brunnea) and northern elephant seal (Mirounga angustirostris) showed inactivating UGT1A6 mutations. A comprehensive literature review of the natural diet of the sampled taxa indicated that all species with defective UGT1A6 were hypercarnivores (>70% dietary animal matter). Furthermore those species with UGT1A6 defects showed evidence for reduced amino acid constraint (increased dN/dS ratios approaching the neutral selection value of 1.0) as compared with species with intact UGT1A6. In contrast, there was no evidence for reduced amino acid constraint for these same species within UGT1A1, the gene encoding the enzyme responsible for detoxification of endogenously generated bilirubin. Our results provide the first evidence suggesting that diet may have played a permissive role in the devolution of a mammalian drug metabolizing enzyme. Further work is needed to establish whether these preliminary findings can be generalized to all Carnivora.

Genetic, individual, and group facilitation of disease resistance in insect societies.

In this review, we provide a current reference on disease resistance in insect societies. We start with the genetics of immunity in the context of behavioral and physiological processes and scale up levels of biological organization until we reach populations. A significant component of this review focuses on Apis mellifera and its role as a model system for studies on social immunity. We additionally review the models that have been applied to disease transmission in social insects and elucidate areas for future study in the field of social immunity.

The energetic costs of stereotyped behavior in the paper wasp, Polistes dominulus.

Polistes wasps engage in many behavioral interactions. Although there has been debate over the meaning of these interactions, these stereotypical behaviors can be used to determine a colony's linear dominance hierarchy. Due to the implicit relationship between behavioral and reproductive dominance, behavioral interactions are commonly used to distinguish the reproductively dominant alpha foundress from the beta foundress. It has been suggested that in order to maintain reproductive control, the alpha foundress is forced to remain at a physiologically constrained activity limit. This, in turn, may allow aggressive interactions to be used as determinants influencing reproductive partitioning between cooperating individuals. Energetic costs can place important limitations on behavior, but the energetic cost of the interactions has not previously been measured. To address this, we measured the CO(2) production of 19 non-nestmate pairs displaying interactive and noninteractive behavior. The rate of energy used during interaction behavior was positively associated with published rankings of aggression. However, our results indicate that interactions are not very energetically costly in Polistes, particularly when compared to the likely cost of foraging. These data suggest that maintaining reproductive dominance is not very energetically expensive for the dominant and that the dominant foundress expends energy at a lower rate than the subordinate foundress.

The ontogeny of immunity: development of innate immune strength in the honey bee (Apis mellifera).

Honey bees (Apis mellifera) are of vital economic and ecological importance. These eusocial animals display temporal polyethism, which is an age-driven division of labor. Younger adult bees remain in the hive and tend to developing brood, while older adult bees forage for pollen and nectar to feed the colony. As honey bees mature, the types of pathogens they experience also change. As such, pathogen pressure may affect bees differently throughout their lifespan. We provide the first direct tests of honey bee innate immune strength across developmental stages. We investigated immune strength across four developmental stages: larvae, pupae, nurses (1-day-old adults), and foragers (22-30 days old adults). The immune strength of honey bees was quantified using standard immunocompetence assays: total hemocyte count, encapsulation response, fat body quantification, and phenoloxidase activity. Larvae and pupae had the highest total hemocyte counts, while there was no difference in encapsulation response between developmental stages. Nurses had more fat body mass than foragers, while phenoloxidase activity increased directly with honey bee development. Immune strength was most vigorous in older, foraging bees and weakest in young bees. Importantly, we found that adult honey bees do not abandon cellular immunocompetence as has recently been proposed. Induced shifts in behavioral roles may increase a colony's susceptibility to disease if nurses begin foraging activity prematurely.

A Darwinian approach to Huntington's disease: subtle health benefits of a neurological disorder.

Huntington's disease (HD) is a neurodegenerative disorder that, unlike most autosomal dominant disorders, is not being selected against. One explanation for the maintenance of the mutant HD allele is that it is transparent to natural selection because disease symptoms typically occur subsequent to an individual's peak reproductive years. While true, this observation does not explain the population-level increase in HD. The increase in HD is at least partly the result of enhanced fitness: HD+ individuals have more offspring than unaffected relatives. This phenomenon has previously been explained as the result of elevated promiscuity of HD+ individuals. For this to be true, disease symptoms must be expressed during the otherwise asymptomatic peak reproductive years and promiscuity must increase offspring production; however, neither prediction is supported by data. Instead, new data suggest that the mutant HD allele bestows health benefits on its carriers. HD+ individuals show elevated levels of the tumor suppressor protein p53 and experience significantly less cancer than unaffected siblings. We hypothesize that the mutant HD allele elevates carriers' immune activity and thus HD+ individuals are, on average, healthier than HD- individuals during reproductive years. As health and reproductive output are positively related, data suggest a counterintuitive relationship: health benefits may lead to an increased prevalence of Huntington's disease.

Transactional skew and assured fitness return models fail to predict patterns of cooperation in wasps.

Cooperative breeders often exhibit reproductive skew, where dominant individuals reproduce more than subordinates. Two approaches derived from Hamilton's inclusive fitness model predict when subordinate behavior is favored over living solitarily. The assured fitness return (AFR) model predicts that subordinates help when they are highly likely to gain immediate indirect fitness. Transactional skew models predict dominants and subordinates "agree" on a level of reproductive skew that induces subordinates to join groups. We show the AFR model to be a special case of transactional skew models that assumes no direct reproduction by subordinates. We use data from 11 populations of four wasp species (Polistes, Liostenogaster) as a test of whether transactional frameworks suffice to predict when subordinate behavior should be observed in general and the specific level of skew observed in cooperative groups. The general prediction is supported; in 10 of 11 cases, transactional models correctly predict presence or absence of cooperation. In contrast, the specific prediction is not consistent with the data. Where cooperation occurs, the model accurately predicts highly biased reproductive skew between full sisters. However, the model also predicts that distantly related or unrelated females should cooperate with low skew. This prediction fails: cooperation with high skew is the observed norm. Neither the generalized transactional model nor the special-case AFR model can explain this significant feature of wasp sociobiology. Alternative, nontransactional hypotheses such as parental manipulation and kin recognition errors are discussed.

Diploid males and their triploid offspring in the paper wasp Polistes dominulus.

Although the hymenopteran sex-determining mechanism generally results in haploid males and diploid females, diploid males can be produced via homozygosity at the sex-determining locus. Diploid males have low fitness because they are effectively sterile or produce presumably sterile triploid offspring. Previously, triploid females were observed in three species of North American Polistes paper wasps, and this was interpreted as indirect evidence of diploid males. Here we report what is, to our knowledge, the first direct evidence: four of five early male-producing Polistes dominulus nests from three populations contained diploid males. Because haploid males were also found, however, the adaptive value of early males cannot be ignored. Using genetic and morphological data from triploid females, we also present evidence that both diploid males and triploid females remain undetected throughout the colony cycle. Consequently, diploid male production may result in a delayed fitness cost for two generations. This phenomenon is particularly relevant for introduced populations with few alleles at the sex-determining locus, but cannot be ignored in native populations without supporting genetic data. Future research using paper wasp populations to test theories of social evolution should explicitly consider the potential impacts of diploid males.

Optimal reproductive-skew models fail to predict aggression in wasps.

Optimal-skew models (OSMs) predict that cooperative breeding occurs as a result of dominants conceding reproductive benefits to subordinates, and that division of reproduction within groups reflects each cooperator's willingness and ability to contest aggressively for dominance. Polistine paper wasps are a leading model system for testing OSMs, and data on reproduction and aggression appear to support OSMs. These studies, however, measure aggression as a single rate rather than by the activity patterns of individuals. This leads to a potential error: if individuals are more likely to receive aggression when active than when inactive, differences in aggression across samples can reflect changes in activity rather than hostility. This study replicates a field manipulation cited as strongly supporting OSMs. We show that fundamentally different conclusions arise when controlling for individual activity states. Our analyses strongly suggest that behaviours classified as 'aggression' in paper wasps are unlikely to function in establishing, maintaining or responding to changes in reproductive skew. This illustrates that OSM tests using aggression or other non-reproductive behaviour as a metric for reproductive partitioning must demonstrate those links rather than assume them.

The function of dart behavior in the paper wasp, Polistes fuscatus.

Dominance behavior in Polistes wasps is a composite trait consisting of various discrete behaviors such as darts, lunges, bites, and mounts. The majority of these behaviors are considered "aggressive", and these aggressive behaviors are considered to form a continuum from mild (e.g., darts) to severe (e.g., falling fights). In this paper we focus on darts, the most common of the dominance behaviors, and investigate their function in un-manipulated post-emergent colonies of the primitively eusocial wasp P. fuscatus. Here we show that darts are correlated with the more severe dominance behaviors, and that dominance ranks do not change with the addition or exclusion of darts. We find no correlation, however, between receiving darts and receiving more severe dominance behaviors. This result suggests that darts are not indicative of aggressive reinforcement of dominance, but rather may serve a different function. Our data suggest that the function of darts is to regulate activity on nests. Both foundresses and workers dart inactive workers significantly more often than by chance, and workers respond to a foundress's (but not a worker's) dart by becoming less inactive. We also found that active workers who receive a dart from either a foundress or worker respond mostly by switching from one activity to another. Thus, our data suggest that darts are not aggressive behaviors, that foundresses use this signal to initiate activity, and that foundresses and workers both use the signal to regulate worker activity.