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.

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.

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.

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.

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.

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.

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.

Bed bug deterrence.

A recent study in BMC Biology has determined that the immature stage of the bed bug (the nymph) signals its reproductive status to adult males using pheromones and thus avoids the trauma associated with copulation in this species. The success of this nymphal strategy of deterrence is instructive. Against the background of increasing problems with bed bugs, this research raises the question whether pheromones might be used to control them. See research article http://www.biomedcentral.com/1741-7007/8/121.

Widespread distribution of knockdown resistance mutations in the bed bug, Cimex lectularius (Hemiptera: Cimicidae), populations in the United States.

We previously reported high deltamethrin resistance in bed bugs, Cimex lectularius, collected from multiple areas of the United States (Romero et al., 2007). Recently, two mutations, the Valine to Leucine mutation (V419L) and the Leucine to Isoleucine mutation (L925I) in voltage-gated sodium channel alpha-subunit gene, had been identified to be responsible for knockdown resistance (kdr) to deltamethrin in bed bugs collected from New York (Yoon et al., 2008). The current study was undertaken to investigate the distribution of these two kdr mutations in 110 bed bug populations collected in the United States. Out of the 17 bed bug populations that were assayed for deltamethrin susceptibility, two resistant populations collected in the Cincinnati area and three deltamethrin-susceptible lab colonies showed neither of the two reported mutations (haplotype A). The remaining 12 populations contained L925I or both V419L and L925I mutations in voltage-gated sodium channel alpha-subunit gene (haplotypes B&C). In 93 populations that were not assayed for deltamethrin susceptibility, 12 contained neither of the two mutations (haplotype A) and 81 contained L925I or V419L or both mutations (haplotypes B-D). Thus, 88% of the bed bug populations collected showed target-site mutations. These data suggest that deltamethrin resistance conferred by target-site insensitivity of sodium channel is widely spread in bed bug populations across the United States.

A chemical signal of offspring quality affects maternal care in a social insect.

Begging signals of offspring are condition-dependent cues that are usually predicted to display information about the short-term need (i.e. hunger) to which parents respond by allocating more food. However, recent models and experiments have revealed that parents, depending on the species and context, may respond to signals of quality (i.e. offspring reproductive value) rather than need. Despite the critical importance of this distinction for life history and conflict resolution theory, there is still limited knowledge of alternative functions of offspring signals. In this study, we investigated the communication between offspring and caring females of the common earwig, Forficula auricularia, hypothesizing that offspring chemical cues display information about nutritional condition to which females respond in terms of maternal food provisioning. Consistent with the prediction for a signal of quality we found that mothers exposed to chemical cues from well-fed nymphs foraged significantly more and allocated food to more nymphs compared with females exposed to solvent (control) or chemical cues from poorly fed nymphs. Chemical analysis revealed significant differences in the relative quantities of specific cuticular hydrocarbon compounds between treatments. To our knowledge, this study demonstrates for the first time that an offspring chemical signal reflects nutritional quality and influences maternal care.

Altered olfactory receptor neuron responsiveness is correlated with a shift in behavioral response in an evolved colony of the cabbage looper moth, Trichoplusia ni.

There is little understanding of how sex pheromone blends might change during speciation events. For the cabbage looper, Trichoplusia ni, there is a mutant laboratory strain that has exhibited characteristics of a shift to a new pheromone blend. Mutant females produce a blend that is significantly different from wild-type females in having a much higher proportion of a minor pheromone component and lower quantity of the major component. Males in this colony have changed over the years to become more broadly tuned and fly upwind equally well to both the wild-type and mutant female pheromone blends. They also exhibit reduced overall sensitivity to pheromone, flying upwind to either blend at a lower success rate than is typical when wild-type males respond to the wild-type blend. Using single-cell recordings, we examined the olfactory receptor neurons (ORNs) of males from evolved and wild-type colonies for evidence of changes in response characteristics that might explain the above-described behavioral evolution. We found that in evolved-colony males the ORNs tuned to the major sex pheromone component exhibited a somewhat lower responsiveness to that compound than the ORNs of wild-type males. In addition, the minor pheromone component, emitted at excessively high rates by mutant females, elicited a drastically reduced ORN responsiveness in evolved-colony males compared to wild-type males. This alteration in ORN responsiveness may be responsible for allowing evolved males to tolerate the excessive amounts of the minor pheromone component in the mutant female blend, which would normally antagonize the upwind flight of unevolved males. Thus, peripheral olfactory alterations have occurred in T. ni males that are correlated with the evolution of the more broadly tuned, but less sensitive, behavioral response profile.

Behavioral responses of the bed bug to insecticide residues.

The recent resurgence of bed bugs, Cimex lectularius L. (Heteroptera: Cimicidae), has increased the demand for information about effective control tactics. Several studies have focused on determining the susceptibility of bed bug populations to insecticides. However, behavioral responses of bed bugs to insecticide residues could influence their efficacy. The behavioral responses of bed bugs to deltamethrin and chlorfenapyr, two commonly used insecticides for bed bug control in the United States, were evaluated. In two-choice tests, grouped insects and individual insects avoided resting on filter paper treated with deltamethrin. Insects did not avoid surfaces treated with chlorfenapyr. Harborages, containing feces and eggs and treated with a deltamethrin-based product, remained attractive to individuals from a strain resistant to pyrethroids. Video recordings of bed bugs indicated that insects increased activity when they contacted sublethal doses of deltamethrin. Insecticide barriers of chlorfenapyr or deltamethrin did not prevent bed bugs from reaching a warmed blood source and acquiring blood meals. We discuss the impact of these responses on bed bug control practices.

Evaluation of piperonyl butoxide as a deltamethrin synergist for pyrethroid-resistant bed bugs.

An understanding of the mechanisms of insecticide resistance in the bed bug, Cimex lectularius L., has the potential to lead to new approaches for the control of resistant populations. We used the cytochrome P450 monooxygenase (P450) inhibitor piperonyl butoxide (PBO) to assess the role of P450s in deltamethrin resistance in three field-collected bed bug strains, LA-1, CIN-1 and WOR-1. In addition, we exposed two highly resistant strains, CIN-1 and WOR-1 (resistance ratio [RR] >2,500-fold), to dry residues of piperonyl butoxide-synergized pyrethroid formulations to determine the utility of synergism by PBO. Piperonyl butoxide synergized deltamethrin in all three strains, but its impact was variable. The synergistic ratio varied from 40 in CIN-1 to 176 in WOR-1. Because the resistance ratio for each strain after piperonyl butoxide treatment was 174 and 39, respectively, our results suggest that P450s have some involvement in deltamethrin resistance, but other resistance mechanisms must be involved as well. No significant synergistic effect of formulated deltamethrin was observed with the addition of synergized pyrethrins or formulated piperonyl butoxide in the CIN-1 strain, but synergism occurred in the WOR-1 strain. Addition of PBO to pyrethroids is not a comprehensive solution to pyrethroid resistance because strains vary in both overall resistance level and the proportion of that resistance attributable to P450s.

Seasonal biology and management of the maple shoot borer, Proteoteras aesculana (Lepidoptera: Tortricidae), in production nurseries.

BACKGROUND: Proteoteras aesculana (Riley), a caterpillar that causes tip dieback and kills the central leader of maples, is an important nursery pest. The authors sought to develop a pheromone lure, use it to clarify the pest's seasonal biology, determine when and where infestations originate and extend the management window. RESULTS: Gas chromatography, electroantennographic detection and trapping showed that (Z)-dodec-8-en-1-ol (Z8-12:OH) is the major sex pheromone component. Flight began in March, peaking in early spring. Larvae developed during April and May, giving rise to moths active in late May and June. No additional larvae were found in maples following that flight. Sporadic moth captures occurred into November, suggesting that adults overwinter. Three parasitoid species, all new host records, were documented. A different tortricid, Episimus tyrius Heinrich, caused leaf-tying damage in late summer. Maples shipped from Oregon were free of shoot borers, refuting speculation that larvae overwinter in buds. One bifenthrin spray applied from just before bud break at onset of moth flight to just after peak flight when shoots had two pairs of leaves reduced infestation by 96-100%. CONCLUSION: Maples are infested soon after planting in Kentucky nurseries via eggs laid in early spring. The temporal window for preventive control is broader than previously thought.

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'.

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.

Insecticide resistance in the bed bug: a factor in the pest's sudden resurgence?

Infestations of the bed bug, Cimex lectularius L. (Heteroptera: Cimicidae), are increasing around the world at an alarming rate and have become a major public health concern. The evolution of insecticide resistance could be a primary factor in explaining this resurgence. Extremely high levels of resistance to two pyrethroid insecticides, deltamethrin and lambda-cyhalothrin, relative to a susceptible colony, were detected in populations collected from human dwellings in Kentucky and Ohio. Offspring of a cross between a resistant and susceptible colony had intermediate susceptibility. Evaluations of populations from across the United States indicate that resistance to pyrethroid insecticides is already widespread. Without the development of new tactics for bed bug management, further escalation of this public health problem should be expected.

Pheromone trap for the eastern tent caterpillar moth.

The discovery that the eastern tent caterpillar Malacosoma americanum (F.) causes mare reproductive loss syndrome (MRLS), and thus has the potential to continue to result in major economic losses to the equine industry of Kentucky, has resulted in an intensive effort to identify practical means to monitor and control this defoliator, including these experiments to optimize a sex pheromone trap for this pest. A pheromone-baited delta trap with a large opening, such as InterceptST Delta, was more effective than other tested traps. Orange delta traps caught more moths than other tested colors. ETC males are caught at all tested heights within the tree canopy. For monitoring flights, setting traps at 1.5 m would allow easy counting of moths. A 9:1 blend of (E,Z)-5,7-dodecadienal (ETC-Ald) and (E,Z)-5,7-dodecadienol (ETC-OH) was most effective in capturing males. Increasing loading doses of a 3:1 blend (Ald:OH) resulted in the capture of increasing numbers of moths, but a 9:1 blend was more effective than 3:1 blend even at a nine-fold lower loading rate. Pheromone-impregnated white septa caught more moths than gray septa at the same loading dose. The advantages and limitations of using pheromone traps for monitoring M. americanum are discussed.

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.