Cooperative policing behaviour regulates reproductive division of labour in a termite.

Reproductive conflicts are common in insect societies where helping castes retain reproductive potential. One of the mechanisms regulating these conflicts is policing, a coercive behaviour that reduces direct reproduction by other individuals. In eusocial Hymenoptera (ants, bees and wasps), workers or the queen act aggressively towards fertile workers, or destroy their eggs. In many termite species (order Blattodea), upon the death of the primary queen and king, workers and nymphs can differentiate into neotenic reproductives and inherit the breeding position. During this process, competition among neotenics is inevitable, but how this conflict is resolved remains unclear. Here, we report a policing behaviour that regulates reproductive division of labour in the eastern subterranean termite, Reticulitermes flavipes. Our results demonstrate that the policing behaviour is a cooperative effort performed sequentially by successful neotenics and workers. A neotenic reproductive initiates the attack of the fellow neotenic by biting and displays alarm behaviour. Workers are then recruited to cannibalize the injured neotenic. Furthermore, the initiation of policing is age-dependent, with older reproductives attacking younger ones, thereby inheriting the reproductive position. This study provides empirical evidence of policing behaviour in termites, which represents a convergent trait shared between eusocial Hymenoptera and Blattodea.

Better than DEET Repellent Compounds Derived from Coconut Oil.

Hematophagous arthropods are capable of transmitting human and animal pathogens worldwide. Vector-borne diseases account for 17% of all infectious diseases resulting in 700,000 human deaths annually. Repellents are a primary tool for reducing the impact of biting arthropods on humans and animals. N,N-Diethyl-meta-toluamide (DEET), the most effective and long-lasting repellent currently available commercially, has long been considered the gold standard in insect repellents, but with reported human health issues, particularly for infants and pregnant women. In the present study, we report fatty acids derived from coconut oil which are novel, inexpensive and highly efficacious repellant compounds. These coconut fatty acids are active against a broad array of blood-sucking arthropods including biting flies, ticks, bed bugs and mosquitoes. The medium-chain length fatty acids from C8:0 to C12:0 were found to exhibit the predominant repellent activity. In laboratory bioassays, these fatty acids repelled biting flies and bed bugs for two weeks after application, and ticks for one week. Repellency was stronger and with longer residual activity than that of DEET. In addition, repellency was also found against mosquitoes. An aqueous starch-based formulation containing natural coconut fatty acids was also prepared and shown to protect pastured cattle from biting flies up to 96-hours in the hot summer, which, to our knowledge, is the longest protection provided by a natural repellent product studied to date.

Managing the risks and rewards of death in eusocial insects.

Eusocial insects frequently face death of colony members as a consequence of living in large groups where the success of the colony is not dependent on the fate of the individual. Whereas death of conspecifics commonly triggers aversion in many group-living species due to risk of pathogens, eusocial insects perform cooperative corpse management. The causes and social context of the death, as well as feeding and nesting ecology of the species, influence the way that corpses are treated. The corpse itself releases cues that dictate the colony's response. As a result, social insects exhibit behavioural responses that promote disease resistance, colony defence and nutrient recycling. Corpse management represents a unique adaption that enhances colony success, and is another factor that has enabled eusocial insects to be so successful. In this review, we summarize the causes of death, the sensory detection of death and corpse management strategies of social insects. In addition, we provide insights into the evolution of behavioural response to the dead and the ecological relevance of corpse management.This article is part of the theme issue 'Evolutionary thanatology: impacts of the dead on the living in humans and other animals'.

Sex-specific inhibition and stimulation of worker-reproductive transition in a termite.

In social insects, the postembryonic development of individuals exhibits strong phenotypic plasticity in response to the environment, thus generating the caste system. Different from eusocial Hymenoptera, in which queens dominate reproduction and inhibit worker fertility, the primary reproductive caste in termites (kings and queens) can be replaced by neotenic reproductives derived from functionally sterile individuals. Feedback regulation of nestmate differentiation into reproductives has been suggested, but the sex specificity remains inconclusive. In the eastern subterranean termite, Reticulitermes flavipes, we tested the hypothesis that neotenic reproductives regulate worker-reproductive transition in a sex-specific manner. With this R. flavipes system, we demonstrate a sex-specific regulatory mechanism with both inhibitory and stimulatory functions. Neotenics inhibit workers of the same sex from differentiating into additional reproductives but stimulate workers of the opposite sex to undergo this transition. Furthermore, this process is not affected by the presence of soldiers. Our results highlight the reproductive plasticity of termites in response to social cues and provide insights into the regulation of reproductive division of labor in a hemimetabolous social insect.

Impact of sublethal exposure to a pyrethroid-neonicotinoid insecticide on mating, fecundity and development in the bed bug Cimex lectularius L. (Hemiptera: Cimicidae).

Sublethal exposure to an insecticide may alter insect feeding, mating, oviposition, fecundity, development, and many other life history parameters. Such effects may have population-level consequences that are not apparent in traditional dose-mortality evaluations. Earlier, we found that a routinely used combination insecticide that includes a pyrethroid and a neonicotinoid (Temprid® SC) had deleterious effects on multiple bed bug (Cimex lectularius, L.) behaviors. Here, we demonstrate that sublethal exposure impacts physiology and reproduction as well. We report that sublethal exposure to Temprid SC has variable aberrant effects on bed bugs depending on the strain, including: a reduction in male mating success and delayed oviposition by females. However, after sublethal exposure, egg hatch rate consistently declined in every strain tested, anywhere from 34%-73%. Conversely, impact on fifth instar eclosion time was not significant. While the strains that we tested varied in their respective magnitude of sublethal effects, taken together, these effects could reduce bed bug population growth. These changes in bed bug behavior and fecundity could lead to improved efficacy of Temprid SC in the field, but recovery of impacted bugs must be considered in future studies. Sublethal effects should not be overlooked when evaluating insecticide efficacy, as it is likely that other products may also have indirect effects on population dynamics that could either aid or inhibit successful management of pest populations.

Social buffering in a eusocial invertebrate: termite soldiers reduce the lethal impact of competitor cues on workers.

While the impact of predator-induced stress on prey has received considerable attention, there has been far less research into the effect of competitors. Cues from aggressive competitors should be particularly likely to evoke behavioral and/or physiological responses, since they may be indicative of both direct (interference) and indirect (exploitative) threats. The danger posed by such competitors, and the "fear" they evoke, should be reduced at lower competitor densities and by the presence of individual conspecifics specialized for defense. We assessed how Reticulitermes flavipes termite workers and soldiers were affected by cues from conspecific nestmates, conspecific non-nestmates, and the heterospecific competitor R. virginicus. Competitor cues altered flavipes worker and soldier behavior, decreasing worker growth and increasing their mortality. The presence of flavipes soldiers largely ameliorated these negative impacts: adding even a single soldier (5% of flavipes individuals) decreased worker mortality by 50-80%. Although worker mortality increased with competitor density, increased soldier densities did not increase the benefit to workers. The small number of soldiers required to substantially alter cue-mediated interactions suggests that this caste, in addition to providing direct defense, also occupies a "keystone role" by providing homeostatic feedback to workers functioning in stressful environments.

Behavioral effects of sublethal exposure to a combination of ß-cyfluthrin and imidacloprid in the bed bug, Cimex lectularius L.

BACKGROUND: Bed bugs (Cimex lectularius) are blood-feeding insect pests with public health relevance. Their rapid evolution of resistance to pyrethroids has prompted a shift to combination products that include both a pyrethroid and neonicotinoid insecticide. Insecticides have both a direct impact on mortality and an indirect effect on behavior. Thus, we assessed the sublethal effects of a widely used combination product containing ß-cyfluthrin (a pyrethroid) and imidacloprid (a neonicotinoid), as unexpected behavioral changes after exposure have been known to affect efficacy of insecticides. RESULTS: We found that bed bugs exposed to sublethal doses of a combination product containing ß-cyfluthrin and imidacloprid did not feed as effectively as untreated bugs. Their locomotion behavior was also reduced. However, aggregation in response to the presence of conspecific harborages was not affected by sublethal exposure. CONCLUSION: Bed bugs exhibit behavioral changes after sublethal exposure to a combination product that could affect pest management choices and outcomes. A reduction in host-finding efficiency and feeding could complement the lethal effects of the insecticide. Alternatively, reduced locomotion following exposure could limit ongoing contact with insecticide deposits. However, an overall reduction in movement indicates that treatments are unlikely to cause dispersal of bugs to adjacent dwellings. © 2016 Society of Chemical Industry.

Quantitative Analysis of Aedes albopictus Movement Behavior Following Sublethal Exposure To Prallethrin.

The pyrethroid prallethrin, an AI in DUET™ (Clarke Mosquito Control, St. Charles, IL), is widely marketed ultra-low volume (ULV) mosquito adulticide. Volatilized prallethrin is intended to stimulate mosquito flight, increasing its adulticide effectiveness. However, field tests using volatilized prallethrin have not produced significant differences in mosquito trap catches, leading to questions regarding prallethrin's behavioral impact efficacy. Thus, we conducted laboratory tests of prallethrin's effect on flight behavior of adult female Asian tiger mosquitoes, Aedes albopictus. Mosquitoes were divided into 3 groups: untreated control, exposed to volatilized prallethrin, and exposed to a liquid spray calibrated to simulate a ULV application at label rates. After exposure, mosquito behavior in an airstream of 0.5 m/sec was recorded and analyzed using motion-tracking software. No significant differences in flight behavior were found between the control and treated mosquitoes exposed to volatilized prallethrin. The ULV-sprayed mosquitoes exhibited a significant increase in the number of flight events, the turning frequency, overall movement speed, and flight speed compared to the control-a significant difference in locomotor stimulation response that would increase exposure to a ULV spray cloud. However, our results showed that volatilization alone was insufficient to increase ULV efficacy in the field and suggested that incorporating a more volatile flight stimulant into ULV adulticides would provide a measurable improvement in mosquito control.

Sex Attractant Pheromone of the Luna Moth, Actias luna (Linnaeus).

Giant silk moths (Lepidoptera: Saturniidae) typically are not well represented as larvae or adults in community level inventories of Lepidoptera, and as a result, little is known about their population dynamics. Furthermore, in recent years, many species of silk moths appear to have experienced population declines. Volatile sex pheromones are powerful sampling tools that can be used in operational conservation and monitoring programs for insects. Here, we describe the identification of the sex attractant pheromone of a giant silk moth, the luna moth Actias luna. Coupled gas chromatography-electroantennographic detection and gas chromatography-mass spectrometric analyses of extracts from pheromone glands of female luna moths supported the identification of (6E,11Z)-6,11-octadecadienal (E6,Z11-18:Ald), (6E)-6-octadecenal (E6-18:Ald), and (11Z)-11-octadecenal (Z11-18:Ald) as the compounds in extracts that elicited responses from antennae of male moths. These identifications were confirmed by synthesis, followed by testing of blends of the synthetic compounds in field trials in Ontario, Canada, and Kentucky, USA. Male moths were attracted to synthetic E6,Z11-18:Ald as a single component. Attraction appeared to be enhanced by addition of E6-18:Ald but not Z11-18:Ald, suggesting that the luna moth pheromone consists of a blend of E6,Z11-18:Ald and E6-18:Ald.

Effects of Ultralow Oxygen and Vacuum Treatments on Bed Bug (Heteroptera: Cimicidae) Survival.

Control of bed bugs is problematic, balancing among efficacy, safety, and cost. In this study, ultralow oxygen (ULO) and vacuum treatments were tested on bed bugs to develop a safer, effective, and environmentally friendly solution to kill bed bugs on infested items. ULO treatments were established by flushing sealed enclosures with nitrogen. All life stages of bed bugs were found to be susceptible to ULO and vacuum treatments, and efficacy of the treatments increased with reduced oxygen levels, increased treatment time, and temperature. In the ULO treatments, 100% mortality of bed bug nymphs and adults and >98% mortality of bed bug eggs were achieved in the 8-h treatment under 0.1% O 2 atmosphere at 30°C. Different levels of vacuum that yielded different oxygen levels were tested on all life stages of bed bugs. The susceptibility of different stages to vacuum treatments increased from nymphs to adults to eggs. Complete control of all life stages was achieved in 12 h under -982 mbar (-29.0 inHg) vacuum at 30°C. This study demonstrated that bed bugs were very susceptible to low oxygen stresses and ULO and vacuum treatments have potential to be used as effective and safe treatments to decontaminate bed bug-infested removable objects.

Insecticide resistance in the bed bug comes with a cost.

Adaptation to new environmental stress is often associated with an alteration of one or more life history parameters. Insecticide resistant populations of insects often have reduced fitness relative to susceptible populations in insecticide free environments. Our previous work showed that three populations of bed bugs, Cimex lectularius L., evolved significantly increased levels of resistance to one product containing both ß-cyfluthrin and imidacloprid insecticides with only one generation of selection, which gave us an opportunity to explore potential tradeoffs between life history parameters and resistance using susceptible and resistant strains of the same populations. Life history tables were compiled by collecting weekly data on mortality and fecundity of bugs from each strain and treatment throughout their lives. Selection led to a male-biased sex ratio, shortened oviposition period, and decreased life-time reproductive rate. Generation time was shortened by selection, a change that represents a benefit rather than a cost. Using these life history characteristics we calculated that there would be a 90% return to pre-selection levels of susceptibility within 2- 6.5 generations depending on strain. The significant fitness costs associated with resistance suggest that insecticide rotation or utilization of non-insecticidal control tactics could be part of an effective resistance management strategy.

Do defensive chemicals facilitate intraguild predation and influence invasion success in ladybird beetles?

Egg predation and cannibalism are believed to be common phenomena among many species of aphidophagous predatory ladybird beetles despite the presence of alkaloid based defensive chemicals in all life stages. We identified defensive chemicals from eggs of three congeneric species, one introduced into North America (Coccinella septempunctata L.), and two native (C. transversoguttata richardsoni Brown, and C. novemnotata Herbst), and examined the effects of ingested defensive chemicals on first instars. Ingested congeneric alkaloids were not toxic to first instars, likely because the three congeners produce the same principal alkaloids, precoccinelline and coccinelline, in similar amounts. First instars of the three congeners accumulated alkaloids ingested through egg cannibalism and congeneric predation. Egg consumption doubled the amount of alkaloids in first instars when they fed on conspecific or congeneric eggs, in comparison to a pea aphid diet. No detrimental effects of ingested congeneric alkaloids on development or survival of first instars were observed among these congeners. Chemical defenses of eggs are therefore not likely to be important in favoring the invasive species, C. septempunctata, in interactions with these native congeneric species. Because the invasive species is the most aggressive predator, having the same types of alkaloids may facilitate disproportionate intraguild predation on native congeners by C. septempunctata thereby potentially enhancing the invasion success of this introduced species.

Antennal olfactory sensilla responses to insect chemical repellents in the common bed bug, Cimex lectularius.

Populations of the common bed bug Cimex lectularius (Hemiptera; Cimicidae), a temporary ectoparasite on both humans and animals, have surged in many developed countries. Similar to other haematophagous arthropods, C. lectularius relies on its olfactory system to detect semiochemicals in the environment, including both attractants and repellents. To elucidate the olfactory responses of the common bed bug to commonly used insect chemical repellents, particularly haematophagous repellents, we investigated the neuronal responses of individual olfactory sensilla in C. lectularius' antennae to 52 insect chemical repellents, both synthetic and botanic. Different types of sensilla displayed highly distinctive response profiles. While C sensilla did not respond to any of the insect chemical repellents, Dγ sensilla proved to be the most sensitive in response to terpene-derived insect chemical repellents. Different chemical repellents elicited neuronal responses with differing temporal characteristics, and the responses of the olfactory sensilla to the insect chemical repellents were dose-dependent, with an olfactory response to the terpene-derived chemical repellent, but not to the non-terpene-derived chemical repellents. Overall, this study furnishes a comprehensive map of the olfactory response of bed bugs to commonly used insect chemical repellents, providing useful information for those developing new agents (attractants or repellents) for bed bug control.

Population variation in and selection for resistance to pyrethroid-neonicotinoid insecticides in the bed bug.

Pyrethroid resistance in bed bugs, Cimex lectularius, has prompted a change to combination products that include a pyrethroid and a neonicotinoid. Ten populations of bed bugs were challenged with two combination products (Temprid SC and Transport GHP). Susceptibility of these populations varied, with the correlated response of the two products indicating cross resistance. We imposed selection on three populations using label rate Temprid, and then reared progeny from unselected and selected strains. All selected strains were significantly less susceptible to Temprid SC than unselected strains. Temprid selected strains were also less susceptible to Transport. The pyrethroid component of Temprid showed a significantly higher LD50 in selected strains, but susceptibility to the neonicotinoid remained unchanged. Taken together these results indicate resistance to combination insecticides is present in field populations at levels that should be of concern, and that short-term selection affecting existing variance in susceptibility can quickly increase resistance.

A tetraene aldehyde as the major sex pheromone component of the promethea moth (Callosamia promethea (Drury)).

The promethea moth Callosamia promethea is one of three species of silkmoths from the genus Callosamia that occur in North America. Cross attraction of males to heterospecific calling females has been observed in the field, and hybrid progeny have been produced by pairing heterospecifics in captivity. These observations suggest that all three species share or have considerable overlap in the sex attractant pheromones produced by females, so that other prezygotic isolating mechanisms, such as diel differences in reproductive activity, limit hybridization in the field. Coupled gas chromatography-electroantennogram detection and gas chromatography- mass-spectrometry analyses of extracts of volatiles collected from female promethea moths supported the identification of (4E,6E,11Z,13Z)-hexadeca-4,6,11,13-tetraenal [(4E,6E,11Z,13Z)-16:Ald] as the compound in extracts that elicited the largest responses from antennae of males. The identification was confirmed by non-selective synthesis of several isomers as analytical standards, and stereoselective synthesis of (4E,6E,11Z,13Z)-16:Ald for testing in field trials. Male moths were strongly attracted to synthetic (4E,6E,11Z,13Z)-16:Ald, suggesting that this compound is the major and possibly the only component of the sex pheromone of these large saturniid moths. Based on the cross-attraction of heterospecifics, it is likely that this is also a major pheromone component of the other two North American Callosamia species as well.

Entrapment of bed bugs by leaf trichomes inspires microfabrication of biomimetic surfaces.

Resurgence in bed bug infestations and widespread pesticide resistance have greatly renewed interest in the development of more sustainable, environmentally friendly methods to manage bed bugs. Historically, in Eastern Europe, bed bugs were entrapped by leaves from bean plants, which were then destroyed; this purely physical entrapment was related to microscopic hooked hairs (trichomes) on the leaf surfaces. Using scanning electron microscopy and videography, we documented the capture mechanism: the physical impaling of bed bug feet (tarsi) by these trichomes. This is distinct from a Velcro-like mechanism of non-piercing entanglement, which only momentarily holds the bug without sustained capture. Struggling, trapped bed bugs are impaled by trichomes on several legs and are unable to free themselves. Only specific, mechanically vulnerable locations on the bug tarsi are pierced by the trichomes, which are located at effective heights and orientations for bed bug entrapment despite a lack of any evolutionary association. Using bean leaves as templates, we microfabricated surfaces indistinguishable in geometry from the real leaves, including the trichomes, using polymers with material properties similar to plant cell walls. These synthetic surfaces snag the bed bugs temporarily but do not hinder their locomotion as effectively as real leaves.

Differential undertaking response of a lower termite to congeneric and conspecific corpses.

Undertaking behaviour is an essential activity in social insects. Corpses are often recognized by a postmortem change in a chemical signature. Reticulitermes flavipes responded to corpses within minutes of death. This undertaking behaviour did not change with longer postmortem time (24 h); however, R. flavipes exhibited distinctively different behaviours toward dead termites from various origins. Corpses of the congeneric species, Reticulitermes virginicus, were buried onsite by workers with a large group of soldiers guarding the burial site due to the risk of interspecific competition; while dead conspecifics, regardless of colony origin, were pulled back into the holding chamber for nutrient recycling and hygienic purposes. The burial task associated with congeneric corpses was coupled with colony defence and involved ten times more termites than retrieval of conspecific corpses. Our findings suggest elicitation of undertaking behaviour depends on the origin of corpses which is associated with different types of risk.

Bed bugs evolved unique adaptive strategy to resist pyrethroid insecticides.

Recent advances in genomic and post-genomic technologies have facilitated a genome-wide analysis of the insecticide resistance-associated genes in insects. Through bed bug, Cimex lectularius transcriptome analysis, we identified 14 molecular markers associated with pyrethroid resistance. Our studies revealed that most of the resistance-associated genes functioning in diverse mechanisms are expressed in the epidermal layer of the integument, which could prevent or slow down the toxin from reaching the target sites on nerve cells, where an additional layer of resistance (kdr) is possible. This strategy evolved in bed bugs is based on their unique morphological, physiological and behavioral characteristics and has not been reported in any other insect species. RNA interference-aided knockdown of resistance associated genes showed the relative contribution of each mechanism towards overall resistance development. Understanding the complexity of adaptive strategies employed by bed bugs will help in designing the most effective and sustainable bed bug control methods.

Effects of juvenile hormone analog formulations on development and reproduction in the bed bug Cimex lectularius (Hemiptera: Cimicidae).

BACKGROUND: Bed bugs (Cimex lectularius L.) have become a common insect pest in urban areas and are often difficult to manage. Eradication is made more problematic by widespread insecticide resistance, raising interest in alternative control products. Juvenile hormone analogs (JHAs) such as methoprene and hydroprene are relatively harmless to non-arthropods and have proved to be effective against other urban insect pests. Two JHA products (Gentrol(®) and Precor(®), Central Life Sciences, Schaumburg, IL) were tested for efficacy against various bed bug stages as direct spray and as dry residue using three bed bug strains. RESULTS: At 1× and 2× the label rate, Precor(®) [active ingredient (S)-methoprene] had no significant effect on the development or fecundity of bed bugs. At 2× the label rate, confinement to residues of Gentrol(®) [active ingredient (S)-hydroprene] had no significant effect, but residues at 3× and 10× the label rate caused a reduction in fecundity and impaired development. Field strains were more susceptible to the reproductive effects of (S)-hydroprene than a long-maintained laboratory strain. CONCLUSIONS: While JHAs are attractive alternatives for pest management because of their inherent safety and distinct mode of action, these JHA formulations would have little impact on bed bug populations without relabeling to allow for higher application rates.