Diverse communication strategies in bees as a window into adaptations to an unpredictable world.

Communication is a fundamental feature of animal societies and helps their members to solve the challenges they encounter, from exploiting food sources to fighting enemies or finding a new home. Eusocial bees inhabit a wide range of environments and they have evolved a multitude of communication signals that help them exploit resources in their environment efficiently. We highlight recent advances in our understanding of bee communication strategies and discuss how variation in social biology, such as colony size or nesting habits, and ecological conditions are important drivers of variation in communication strategies. Anthropogenic factors, such as habitat conversion, climate change, or the use of agrochemicals, are changing the world bees inhabit, and it is becoming clear that this affects communication both directly and indirectly, for example by affecting food source availability, social interactions among nestmates, and cognitive functions. Whether and how bees adapt their foraging and communication strategies to these changes represents a new frontier in bee behavioral and conservation research.

Effects of native forest and human-modified land covers on the accumulation of toxic metals and metalloids in the tropical bee Tetragonisca angustula.

The intensive shift on land cover by anthropogenic activities have led to changes in natural habitats and environmental contamination, which can ultimately impact and threat biodiversity and ecosystem services, such as pollination. The aim of this study was to evaluate the effect of native forest and human-modified land covers on the concentrations of chemical elements accumulated in the neotropical pollinator bee T. angustula. Eight landscapes, within an Ecological Corridor in the State of São Paulo, Brazil, with gradients of forest cover, spatial heterogeneity and varying land covers were used as sampling unities. Bees collected in traps or through actives searches had the concentration of 21 chemical elements determined by ICP-MS. Results show a beneficial effect of forested areas on the concentrations of some well-known toxic elements accumulated in bees, such as Hg, Cd, and Cr. Multivariate Redundancy Analysis (RDA) suggests road as the most important driver for the levels of Cr, Hg, Sb, Al, U, As, Pb and Pt and bare soil, pasture and urban areas as the landscape covers responsible for the concentrations of Zn, Cd, Mn, Mg, Ba and Sr in bees. The results reinforce the potential use of T. angustula bees as bioindicators of environmental quality and also show that these organisms are being directly affected by human land use, offering potential risks for the Neotropical ecosystem. Our study sheds light on how land covers (native forest and human-modified) can influence the levels of contaminants in insects within human-dominated landscapes. The generation of predictions of the levels of toxic metals and metalloids based on land use can both contribute to friendly farming planning as well as to support public policy development on the surrounding of protected areas and biodiversity conservation hotspots.

Diploid Male Production Results in Queen Death in the Stingless Bee Scaptotrigona depilis.

As in most Hymenoptera, the eusocial stingless bees (Meliponini) have a complementary sex determination (CSD) system. When a queen makes a "matched mating" with a male that shares a CSD allele with her, half of their diploid offspring are diploid males rather than females. Matched mating imposes a cost, since diploid male production reduces the colony workforce. Hence, adaptations preventing the occurrence or attenuating its effects are likely to arise. Here we provide clear evidence that in the stingless bee Scaptotrigona depilis, the emergence of diploid males induces queen death, and this usually occurs within 10-20 days of the emergence of diploid male offspring from their pupae. Queens that have not made a matched mating die when introduced into a colony in which diploid males are emerging. This shows that the adult diploid males, and not the queen that has made a matched mating herself, are the proximate cause of queen death. Analysis of the cuticular hydrocarbon profiles of adult haploid and diploid males shows six compounds with significant differences. Moreover, the diploid and haploid males only acquire distinct cuticular hydrocarbon profiles 10 days after emergence. Our data shows that the timing of queen death occurs when the cuticular hydrocarbons of haploid and diploid males differ significantly, suggesting that these chemical differences could be used as cues or signals to trigger queen death.

Beekeeping practices and geographic distance, not land use, drive gene flow across tropical bees.

Across the globe, wild bees are threatened by ongoing natural habitat loss, risking the maintenance of plant biodiversity and agricultural production. Despite the ecological and economic importance of wild bees and the fact that several species are now managed for pollination services worldwide, little is known about how land use and beekeeping practices jointly influence gene flow. Using stingless bees as a model system, containing wild and managed species that are presumed to be particularly susceptible to habitat degradation, here we examine the main drivers of tropical bee gene flow. We employ a novel landscape genetic approach to analyse data from 135 populations of 17 stingless bee species distributed across diverse tropical biomes within the Americas. Our work has important methodological implications, as we illustrate how a maximum-likelihood approach can be applied in a meta-analysis framework to account for multiple factors, and weight estimates by sample size. In contrast to previously held beliefs, gene flow was not related to body size or deforestation, and isolation by geographic distance (IBD) was significantly affected by management, with managed species exhibiting a weaker IBD than wild ones. Our study thus reveals the critical importance of beekeeping practices in shaping the patterns of genetic differentiation across bee species. Additionally, our results show that many stingless bee species maintain high gene flow across heterogeneous landscapes. We suggest that future efforts to preserve wild tropical bees should focus on regulating beekeeping practices to maintain natural gene flow and enhancing pollinator-friendly habitats, prioritizing species showing a limited dispersal ability.

The origin and evolution of queen and fertility signals in Corbiculate bees.

BACKGROUND: In social Hymenoptera (ants, bees and wasps), various chemical compounds present on the cuticle have been shown to act as fertility signals. In addition, specific queen-characteristic hydrocarbons have been implicated as sterility-inducing queen signals in ants, wasps and bumblebees. In Corbiculate bees, however, the chemical nature of queen-characteristic and fertility-linked compounds appears to be more diverse than in ants and wasps. Moreover, it remains unknown how queen signals evolved across this group and how they might have been co-opted from fertility signals in solitary ancestors. RESULTS: Here, we perform a phylogenetic analysis of fertility-linked compounds across 16 species of solitary and eusocial bee species, comprising both literature data as well as new primary data from a key solitary outgroup species, the oil-collecting bee Centris analis, and the highly eusocial stingless bee Scaptotrigona depilis. Our results demonstrate the presence of fertility-linked compounds belonging to 12 different chemical classes. In addition, we find that some classes of compounds (linear and branched alkanes, alkenes, esters and fatty acids) were already present as fertility-linked signals in the solitary ancestors of Corbiculate bees, while others appear to be specific to certain species. CONCLUSION: Overall, our results suggest that queen signals in Corbiculate bees are likely derived from ancestral fertility-linked compounds present in solitary bees that lacked reproductive castes. These original fertility-linked cues or signals could have been produced either as a by-product of ovarian activation or could have served other communicative purposes, such as in mate recognition or the regulation of egg-laying.

Antibacterial Potential of 2.5% Sodium Hypochlorite in Distinct Irrigation Protocols on Enterococcus faecalis Biofilm.

OBJECTIVE: The aim of this study was to evaluate the effect of irrigation methods on antibacterial potential of 2.5% NaOCl on Enterococcus faecalis biofilm. MATERIALS AND METHODS: Enterococcus faecalis biofilms were prepared during 60 days on 48 human root canals and randomized into control and experimental groups using positive and negative pressure irrigation. Bacterial growth was analyzed using turbidity of culture medium followed by UV spectrophotometry, and scanning electron microscopy (SEM) analyses were performed. Mean and standard deviations were used for evaluate the mean optical densities associated to the number of bacteria present culture, and Scheirer-Ray-Hare (an extension of the Kruskal-Wallis test) and Tamhane test to analyze the SEM images in the groups and thirds. Significance was set at 5%. RESULTS: Enterococcus faecalis was still present after root canal cleaning regardless of irrigation methods or bacterial identification methods. CONCLUSION: Positive and negative pressure irrigation protocols using 2.5% NaOCl show a similar capacity to reduce E. faecalis in infected root canals.

Monogamy in large bee societies: a stingless paradox.

High genetic diversity is important for the functioning of large insect societies. Across the social Hymenoptera (ants, bees, and wasps), species with the largest colonies tend to have a high colony-level genetic diversity resulting from multiple queens (polygyny) or queens that mate with multiple males (polyandry). Here we studied the genetic structure of Trigona spinipes, a stingless bee species with colonies an order of magnitude larger than those of polyandrous honeybees. Genotypes of adult workers and pupae from 43 nests distributed across three Brazilian biomes showed that T. spinipes colonies are usually headed by one singly mated queen. Apart from revealing a notable exception from the general incidence of high genetic diversity in large insect societies, our results reinforce previous findings suggesting the absence of polyandry in stingless bees and provide evidence against the sperm limitation hypothesis for the evolution of polyandry. Stingless bee species with large colonies, such as T. spinipes, thus seem promising study models to unravel alternative mechanisms to increase genetic diversity within colonies or understand the adaptive value of low genetic diversity in large insect societies.

Toward greater realism in inclusive fitness models: the case of caste fate conflict in insect societies.

In the field of social evolution, inclusive fitness theory has been successful in making a wide range of qualitative predictions on expected patterns of cooperation and conflict. Nevertheless, outside of sex ratio theory, inclusive fitness models that make accurate quantitative predictions remain relatively rare. Past models dealing with caste fate conflict in insect societies, for example, successfully predicted that if female larvae can control their own caste fate, an excess should opt to selfishly develop as queens. Available models, however, were unable to accurately predict levels of queen production observed in Melipona bees-a genus of stingless bees where caste is self-determined-as empirically observed levels of queen production are approximately two times lower than the theoretically predicted ones. Here, we show that this discrepancy can be resolved by explicitly deriving the colony-level cost of queen overproduction from a dynamic model of colony growth, requiring the incorporation of parameters of colony growth and demography, such as the per-capita rate at which new brood cells are built and provisioned, the percentage of the queen's eggs that are female, costs linked with worker reproduction and worker mortality. Our revised model predicts queen overproduction to more severely impact colony productivity, resulting in an evolutionarily stable strategy that is approximately half that of the original model, and is shown to accurately predict actual levels of queen overproduction observed in different Melipona species. Altogether, this shows how inclusive fitness models can provide accurate quantitative predictions, provided that costs and benefits are modeled in sufficient detail and are measured precisely.

Prospective monitoring of patients undergoing radiotherapy during COVID-19.

OBJECTIVE: The objective of this study was to evaluate the quality of life of consecutive patients undergoing radiotherapy during the coronavirus disease 2019 pandemic at a private hospital in Southern Brazil from September 2020 to September 2021. METHODS: This study was approved by the Research Ethics Board under project number 112 on April 17, 2020, and it was a prospective descriptive cohort study conducted in a Brazilian radiotherapy department from September 2020 to September 2021. It involved the weekly administration of the European Organisation for Research and Treatment of Cancer Questionnaire Core 30 questionnaires via telephone to consecutively assess patients with pathology-proven cancer diagnoses. These questionnaires captured both demographic data and patients' concerns related to the pandemic, providing a comprehensive overview of their quality of life during radiotherapy treatment. RESULTS: In this study, 141 patients were analyzed, predominantly female (69.5%) with an average age of 61 years. Breast and prostate were the most treated sites, accounting for 51 and 19% of cases, respectively. The majority of treatments lasted between 3 and 5 weeks (73.77%). A small fraction (4.26%) tested positive for coronavirus disease 2019. The findings also highlighted a relatively high quality of life, with mean global scores of 77.95 and emotional functioning scores of 87.53, indicating maintained well-being during treatment. CONCLUSIONS: Oncological patients continuing radiotherapy at our center during the pandemic experienced a low coronavirus disease 2019 infection rate and maintained a high quality of life with minimal emotional distress throughout their treatment period.

Bourgeois behavior and freeloading in the colonial orb web spider Parawixia bistriata (Araneae, Araneidae).

Spiders of the tropical American colonial orb weaver Parawixia bistriata form a communal bivouac in daytime. At sunset, they leave the bivouac and construct individual, defended webs within a large, communally built scaffolding of permanent, thick silk lines between trees and bushes. Once spiders started building a web, they repelled other spiders walking on nearby scaffolding with a "bounce" behavior. In nearly all cases (93%), this resulted in the intruder leaving without a fight, akin to the "bourgeois strategy," in which residents win and intruders retreat without escalated contests. However, a few spiders (6.5%) did not build a web due to lack of available space. Webless spiders were less likely to leave when bounced (only 42% left) and instead attempted to "freeload," awaiting the capture of prey items in nearby webs. Our simple model shows that webless spiders should change their strategy from bourgeois to freeloading satellite as potential web sites become increasingly occupied.

Towards greater realism in inclusive fitness models: the case of worker reproduction in insect societies.

The conflicts over sex allocation and male production in insect societies have long served as an important test bed for Hamilton's theory of inclusive fitness, but have for the most part been considered separately. Here, we develop new coevolutionary models to examine the interaction between these two conflicts and demonstrate that sex ratio and colony productivity costs of worker reproduction can lead to vastly different outcomes even in species that show no variation in their relatedness structure. Empirical data on worker-produced males in eight species of Melipona bees support the predictions from a model that takes into account the demographic details of colony growth and reproduction. Overall, these models contribute significantly to explaining behavioural variation that previous theories could not account for.

Factors influencing survival duration and choice of virgin queens in the stingless bee Melipona quadrifasciata.

In Melipona quadrifasciata, about 10% of the females develop into queens, almost all of which are killed. Occasionally, a new queen replaces or supersedes the mother queen or heads a new colony. We investigated virgin queen fate in queenright and queenless colonies to determine the effects of queen behaviour, body mass, nestmate or non-nestmate status, queenright or queenless colony status, and, when queenless, the effect of the time a colony had been queenless, on survival duration and acceptance. None of 220 virgin queens observed in four observation hives ever attacked another virgin queen nor did any of 88 virgin queens introduced into queenright colonies ever attack the resident queen. A new queen was only accepted in a queenless colony. Factors increasing survival duration and acceptance of virgin queens were to emerge from its cell at 2 h of queenlessness, to hide, and to avoid fights with workers. In this way, a virgin queen was more likely to be available when a colony chooses a new queen, 24-48 h after resident queen removal. Running, walking or resting, antennating or trophallaxis, played little or no role, as did the factors body mass or nestmate. "Queen choice" took about 2 h during which time other virgin queens were still being killed by workers. During this agitated process, the bees congregated around the new queen. She inflated her abdomen and some of the workers deposited a substance on internal nest surfaces including the glass lid of the observation hive.

A predatory social wasp does not avoid nestmates contaminated with a fungal biopesticide.

Fungus-based biopesticides have been used worldwide for crop pest control as a safer alternative to chemical pesticides such as neonicotinoids. Both agrochemicals can be lethal and may also trigger side effects on the behavioral traits of non-target social insects, which play a crucial role in providing essential biological pest control services in agroecosystems. Here, we evaluated whether a commercial formulation of the entomopathogenic fungus Beauveria bassiana or the neonicotinoid imidacloprid causes mortality in foragers of Mischocyttarus metathoracicus. These social wasps are natural enemies of caterpillars and other herbivorous insects and inhabit both urban and agricultural environments in Brazil. We also tested whether wasps discriminate between biopesticide-exposed and unexposed conspecifics. Through a combination of laboratory (survival assay) and field experiments (lure presentation), along with chemical analyses (cuticular hydrocarbon profiles), we showed that topic exposure to the label rate of each pesticide causes a lethal effect, with the biopesticide exhibiting a slower effect. Moreover, wasps do not discriminate biopesticide-exposed from unexposed conspecifics, likely because of the similarity of their cuticular chemical profiles 24 h after exposure. Overall, the delayed lethal time at the individual level, combined with the indistinctive chemical cues of exposure and the lack of discrimination by conspecifics suggests that the fungal biopesticide may ultimately pose a threat to the colony survival of this predatory wasp.

Laboratory Risk Assessment of Three Entomopathogenic Fungi Used for Pest Control toward Social Bee Pollinators.

The use of fungal-based biopesticides to reduce pest damage and protect crop quality is often considered a low-risk control strategy. Nevertheless, risk assessment of mycopesticides is still needed since pests and beneficial insects, such as pollinators, co-exist in the same agroecosystem where mass use of this strategy occurs. In this context, we evaluated the effect of five concentrations of three commercial entomopathogenic fungi, Beauveria bassiana, Metarhizium anisopliae, and Cordyceps fumosorosea, by direct contact and ingestion, on the tropical stingless bees Scaptotrigona depilis and Tetragonisca angustula, temperate bee species, the honey bee Apis mellifera, and the bumble bee Bombus terrestris, at the individual level. Furthermore, we studied the potential of two infection routes, either by direct contact or ingestion. In general, all three fungi caused considerable mortalities in the four bee species, which differed in their response to the different fungal species. Scaptotrigona depilis and B. terrestris were more susceptible to B. bassiana than the other fungi when exposed topically, and B. terrestris and A. mellifera were more susceptible to M. anisopliae when exposed orally. Interestingly, increased positive concentration responses were not observed for all fungal species and application methods. For example, B. terrestris mortalities were similar at the lowest and highest fungal concentrations for both exposure methods. This study demonstrates that under laboratory conditions, the three fungal species can potentially reduce the survival of social bees at the individual level. However, further colony and field studies are needed to elucidate the susceptibility of these fungi towards social bees to fully assess the ecological risks.

Side effects of a fungus-based biopesticide on stingless bee guarding behaviour.

Pathogenic fungi have been used worldwide to control crop pests and are assumed to pose negligible threats to the survival of pollinators. Although eusocial stingless bees provide essential pollination services and might be exposed to these biopesticides in tropical agroecosystems, there is a substantial knowledge gap regarding the side effects of fungal pathogens on behavioural traits that are crucial for colony functioning, such as guarding behaviour. Here, we evaluated the effect of Beauveria bassiana on the sophisticated kin recognition system of Tetragonisca angustula, a bee with morphologically specialized entrance guards. By combining behavioural assays and chemical analyses, we show that guards detect pathogen-exposed nestmates, preventing them from accessing nests. Furthermore, cuticular profiles of pathogen-exposed foragers contained significantly lower amounts of linear alkanes than the unexposed ones. Such chemical cues associated with fungal conidia may potentially trigger aggression towards pathogen-exposed bees, preventing pathogen spread into and among colonies. This is the first demonstration that this highly abundant native bee seems to respond in a much more adaptive way to a potentially infectious threat, outweighing the costs of losing foraging workforce when reducing the chances of fungal pathogen outbreaks within their colonies, than honeybees do.

Effect of low-power diode laser on infected root canals.

This study evaluated the effect of photodynamic therapy (PDT) on infected root canals. Twenty-one human teeth were selected, and 18 were infected by E. faecalis for 60 days. The antimicrobial strategies tested were: G1. Root canal preparation (RCP) using Niquel-Titanium (NiTi) rotary instruments, 2.5% NaOCl, and final irrigation with 17% EDTA, followed by PDT with methylene blue photosensitizer and laser diode low power; G2. RCP using stainless steel files and the same irrigation and PDT protocols as G1; G3. Same RCP protocol as G1 without PDT; G4. Only irrigation with 2.5% NaOCl; G5. Same PDT protocol as G1 without RCP; G6. Negative control; G7. Positive control. Samples for microbiological tests were collected initially (S1), after RCP (S2), and after PDT (S3). Subsequently, the roots were sectioned and prepared for Scanning Electron Microscopy (SEM) analysis. Bacterial growth was analyzed according to the turbidity of the culture medium, followed by spectrophotometric optical density (nm). The effect of PDT on the dentinal structure was evaluated at magnifications 1,600X and 5,000X and described qualitatively. The Wilcoxon test was used for the comparisons from the same specimens, and the Mann-Whitney test was used to compare groups ((=5%). Bacteria were found in all experimental groups' microbiological samples (S1, S2 and S3). The optical density of culture media was lower in S2 than in S1 of G1, 2, 3, and 4 (p> 0.05). After PDT (S3) in G1 and 2, there was an additional reduction in optical density of the culture medium, respectively (p>0.05). In Group 5, the analysis of culture media at S2 revealed an increase in optical density compared to S1(p>0.05). In SEM images of G1, 2, and 5, dentin with melting and recrystallization areas were evidenced. After preparation of the root canal with the rotary system or manually associated with 2.5% NaOCl, PDT was not able to completely eliminate E. faecalis present in the root canal.

Intraspecific queen parasitism in a highly eusocial bee.

Insect societies are well-known for their advanced cooperation, but their colonies are also vulnerable to reproductive parasitism. Here, we present a novel example of an intraspecific social parasitism in a highly eusocial bee, the stingless bee Melipona scutellaris. In particular, we provide genetic evidence which shows that, upon loss of the mother queen, many colonies are invaded by unrelated queens that fly in from unrelated hives nearby. The reasons for the occurrence of this surprising form of social parasitism may be linked to the fact that unlike honeybees, Melipona bees produce new queens in great excess of colony needs, and that this exerts much greater selection on queens to seek alternative reproductive options, such as by taking over other nests. Overall, our results are the first to demonstrate that queens in highly eusocial bees can found colonies not only via supersedure or swarming, but also by infiltrating and taking over other unrelated nests.

Withdrawing Ixekizumab in Patients With Psoriatic Arthritis Who Achieved Minimal Disease Activity: Results From a Randomized, Double-Blind Withdrawal Study.

OBJECTIVE: To evaluate the effect of withdrawing ixekizumab in patients with psoriatic arthritis (PsA) in whom minimal disease activity (MDA) has been achieved after open-label ixekizumab treatment. METHODS: SPIRIT-P3 was a multicenter, randomized, double-blind withdrawal study of biologic treatment-naive adult patients with PsA who were treated with open-label ixekizumab for 36 weeks (160 mg at week 0, then 80 mg every 2 weeks). Patients in whom MDA was sustained for >3 consecutive months were randomized 1:1, between weeks 36 and 64, to undergo blinded withdrawal of ixekizumab treatment (placebo) or to continue ixekizumab treatment every 2 weeks up to week 104. The primary efficacy end point was time to relapse (loss of MDA) for randomized patients. Patients who experienced a relapse were re-treated with ixekizumab every 2 weeks up to week 104. RESULTS: A total of 394 patients were enrolled and received open-label ixekizumab every 2 weeks. Of those patients, 158 (40%) achieved sustained MDA and were randomized to undergo withdrawal of ixekizumab treatment (placebo every 2 weeks; n = 79) or to continue ixekizumab treatment every 2 weeks (n = 79). Disease relapse occurred more rapidly with treatment withdrawal (median 22.3 weeks [95% confidence interval (95% CI) 16.1-28.3]) compared to those who continued treatment with ixekizumab (median not estimable; P < 0.0001). Sixty-seven patients (85%) compared to 30 patients (38%) experienced relapse in the placebo group and the continued treatment group, respectively. Median time to achieving MDA again with re-treatment was 4.1 weeks (95% CI 4.1-4.3); in 64 of 67 patients (96%) who experienced relapse with treatment withdrawal, MDA was achieved again with re-treatment. Safety was consistent with the known safety profile for ixekizumab. CONCLUSION: Continued ixekizumab therapy is superior to ixekizumab withdrawal in maintaining low disease activity in biologic treatment-naive patients with PsA. Re-treatment with ixekizumab following a relapse may restore disease control in cases of treatment interruption.

Prospective monitoring of patients undergoing radiotherapy during COVID-19

SUMMARY OBJECTIVE: The objective of this study was to evaluate the quality of life of consecutive patients undergoing radiotherapy during the coronavirus disease 2019 pandemic at a private hospital in Southern Brazil from September 2020 to September 2021. METHODS: This study was approved by the Research Ethics Board under project number 112 on April 17, 2020, and it was a prospective descriptive cohort study conducted in a Brazilian radiotherapy department from September 2020 to September 2021. It involved the weekly administration of the European Organisation for Research and Treatment of Cancer Questionnaire Core 30 questionnaires via telephone to consecutively assess patients with pathology-proven cancer diagnoses. These questionnaires captured both demographic data and patients' concerns related to the pandemic, providing a comprehensive overview of their quality of life during radiotherapy treatment. RESULTS: In this study, 141 patients were analyzed, predominantly female (69.5%) with an average age of 61 years. Breast and prostate were the most treated sites, accounting for 51 and 19% of cases, respectively. The majority of treatments lasted between 3 and 5 weeks (73.77%). A small fraction (4.26%) tested positive for coronavirus disease 2019. The findings also highlighted a relatively high quality of life, with mean global scores of 77.95 and emotional functioning scores of 87.53, indicating maintained well-being during treatment. CONCLUSIONS: Oncological patients continuing radiotherapy at our center during the pandemic experienced a low coronavirus disease 2019 infection rate and maintained a high quality of life with minimal emotional distress throughout their treatment period.

Organization enhances collective vigilance in the hovering guards of Tetragonisca angustula bees.

One benefit of group living is vigilance against predators. Previous studies have investigated the group size effect, where individual vigilance decreases as group size increases without reducing the overall ability of the group to detect predators. However, there has been comparatively little research on whether the positioning of individuals can improve the collective vigilance of the group. We studied the coordination of vigilance and its effect on predator detection in the eusocial bee Tetragonisca angustula. Nests are defended by hovering guards that detect and intercept intruders before they reach the nest entrance, in addition to those that stand upon it. We show that hovering guards are positioned nonrandomly, with a strong tendency for equal numbers on both sides of the entrance. This organization increases the collective vigilance of the guard group, as groups distributed in an even ratio, either side of the entrance, have a greater collective field of view than groups that deviate from an even ratio. Finally, we use a bioassay to show that when guards are on both sides of the entrance, their ability to detect intruders before they reach the entrance increases. Overall, our results provide strong evidence that vigilance is coordinated and that this improves nest defense. Although other group-living animals are often selfish in their individual vigilance behaviors and face competing time constraints such as foraging, the altruistic nature of eusocial insect workers has probably facilitated the evolution of coordinated vigilance, as documented here in T. angustula.