Developmental plasticity of the locomotor activity rhythm of Drosophila melanogaster

We used four replicate outbred populations of Drosophila melanogaster to investigate whether the light regimes experienced during the pre-adult (larval and pupal) and early adult stages influence the free-running period (τ_DD) of the circadian locomotor activity rhythm of adult flies. In a series of two experiments four different populations of flies were raised from egg to eclosion in constant light (LL), in light/dark (LD) 12:12 h cycle, and in constant darkness (DD). In the first experiment the adult male and female flies were directly transferred into DD and their locomotor activity was monitored, while in the second experiment the locomotor activity of the emerging adult flies was first assayed in LD 12:12 h for 15 days and then in DD for another 15 days. The τ_DD of the locomotor activity rhythm of flies that were raised in all the three light regimes, LL, LD 12:12 h and in DD was significantly different from each other. The τ_DD of the locomotor activity rhythm of the flies, which were raised in DD during their pre-adult stages, was significantly shorter than that of flies that were raised as pre-adults in LL regime, which in turn was significantly shorter than that of flies raised in LD 12:12 h regime. This pattern was consistent across both the experiments. The results of our experiments serve to emphasise the fact that in order to draw meaningful inferences about circadian rhythm parameters in insects, adequate attention should be paid to control and specify the environment in which pre-adult rearing takes place. The pattern of pre-adult and early adult light regime effects that we see differs from that previously observed in studies of mutant strains of D. melanogaster, and therefore, also points to the potential importance of inter-strain differences in the response of circadian organisation to external influences.     

Mark-Release-Recapture Reveals Extensive Movement of Bed Bugs (Cimex lectularius L.) within and between Apartments

Understanding movement and dispersal of the common bed bug (Cimex lectularius L.) under field conditions is important in the control of infestations and for managing the spread of bed bugs to new locations. We investigated bed bug movement within and between apartments using mark-release-recapture (m-r-r) technique combined with apartment-wide monitoring using pitfall-style interceptors. Bed bugs were collected, marked, and released in six apartments. The distribution of marked and unmarked bed bugs in these apartments and their 24 neighboring units were monitored over 32 days. Extensive movement of marked bed bugs within and between apartments occurred regardless of the number of bed bugs released or presence/absence of a host. Comparison of marked and unmarked bed bug distributions confirms that the extensive bed bug activity observed was not an artifact of the m-r-r technique used. Marked bed bugs were recovered in apartments neighboring five of six m-r-r apartments. Their dispersal rates at 14 or 15 d were 0.0–5.0%. The estimated number of bed bugs per apartment in the six m-r-r apartments was 2,433–14,291 at 4–7 d after release. Longevity of bed bugs in the absence of a host was recorded in a vacant apartment. Marked large nymphs (3^rd– 5^th instar), adult females, and adult males continued to be recovered up to 57, 113, and 134 d after host absence, respectively. Among the naturally existing unmarked bed bugs, unfed small nymphs (1^st– 2^nd instar) were recovered up to 134 d; large nymphs and adults were still found at 155 d when the study ended. Our findings provide important insight into the behavioral ecology of bed bugs in infested apartments and have significant implications in regards to eradication programs and managing the spread of bed bugs within multi-occupancy dwellings.     

Host Chemical Footprints Induce Host Sex Discrimination Ability in Egg Parasitoids

Trissolcus egg parasitoids, when perceiving the chemical footprints left on a substrate by pentatomid host bugs, adopt a motivated searching behaviour characterized by longer searching time on patches were signals are present. Once in contact with host chemical footprints, Trissolcus wasps search longer on traces left by associated hosts rather than non-associated species, and, in the former case, they search longer on traces left by females than males. Based on these evidences, we hypothesized that only associated hosts induce the ability to discriminate host sex in wasps. To test this hypothesis we investigated the ability of Trissolcus basalis, T. brochymenae, and Trissolcus sp. to distinguish female from male Nezara viridula, Murgantia histrionica, and Graphosoma semipunctatum footprints. These three pentatomid bugs were selected according to variable association levels. Bioassays were conducted on filter paper sheets, and on Brassica oleracea (broccoli) leaves. The results confirmed our hypothesis showing that wasps spent significantly more time on female rather than male traces left by associated hosts on both substrates. No differences were observed in the presence of traces left by non-associated hosts. The ecological consequences for parasitoid host location behaviour are discussed.     

Role of Vision and Mechanoreception in Bed Bug,Cimex lectularius L. Behavior

The role of olfactory cues such as carbon dioxide, pheromones, and kairomones in bed bug, Cimex lectularius L. behavior has been demonstrated. However, the role of vision and mechanoreception in bed bug behavior is poorly understood. We investigated bed bug vision by determining their responses to different colors, vertical objects, and their ability to detect colors and vertical objects under low and complete dark conditions. Results show black and red paper harborages are preferred compared to yellow, green, blue, and white harborages. A bed bug trapping device with a black or red exterior surface was significantly more attractive to bed bugs than that with a white exterior surface. Bed bugs exhibited strong orientation behavior toward vertical objects. The height (15 vs. 30 cm tall) and color (brown vs. black) of the vertical object had no significant effect on orientation behavior of bed bugs. Bed bugs could differentiate color and detect vertical objects at very low background light conditions, but not in complete darkness. Bed bug preference to different substrate textures (mechanoreception) was also explored. Bed bugs preferred dyed tape compared to painted tape, textured painted plastic, and felt. These results revealed that substrate color, presence of vertical objects, and substrate texture affect host-seeking and harborage-searching behavior of bed bugs. Bed bugs may use a combination of vision, mechanoreception, and chemoreception to locate hosts and seek harborages.     

Questing activity in bed bug populations: male and female responses to host signals

A large‐arena bioassay is used to examine sex differences in spatiotemporal patterns of bed bug Cimex lectularius L. behavioural responses to either a human host or CO₂ gas. After release in the centre of the arena, 90% of newly‐fed bed bugs move to hiding places in the corners within 24 h. They require 3 days to settle down completely in the arena, with generally low activity levels and the absence of responses to human stimuli for 5 days. After 8–9 days, persistent responses can be recorded. Sex differences are observed, in which females are more active during establishment, respond faster after feeding, expose themselves more than males during the daytime, and respond more strongly to the host signal. The number of bed bugs that rest in harbourages is found to vary significantly according to light setting and sex. Both sexes stay inside harbourages more in daylight compared with night, and males hide more than females during the daytime but not during the night. The spatial distribution of the bed bugs is also found to change with the presence of CO₂, and peak aggregation around the odour source is observed after 24 min. Both male and female bed bugs move from hiding places or the border of the arena toward the centre where CO₂ is released. Peak responses are always highest during the night. Bed bug behaviour and behaviour‐regulating features are discussed in the context of control methods.     

Circadian system responses to nocturnal and diurnal hosts in the kissing bug,Triatoma infestans

ABSTRACT

Insects express diverse behavioral rhythms synchronized to environmental cycles. While circadian entrainment to light–dark cycles is ubiquitous in living organisms, synchronization to non-photic cycles may be critical for hematophagous bugs that depend on rhythmic hosts. The purpose was to determine whether Triatoma infestans are capable of synchronizing to the circadian rhythms of potential hosts with temporally distinct activity patterns; and, if so, if this synchronization occurs through masking or entrainment. Precise synchronization with the food source may be critical for the insects’ survival due to the specific predatory or defensive nature of each host. Kissing bugs were housed in a compartment in constant dark, air-flow-connected to another compartment with a nocturnal or a diurnal host; both hosts were synchronized to a light–dark cycle. The activity rhythms of kissing bugs were modulated by the daily activity rhythms of the vertebrates. Effects were a decrease in the endogenous circadian period, independent of the host being nocturnal or diurnal; in some cases relative coordination occurred and in others synchronization was clearly achieved. Moreover, splitting and bimodality arose, phenomena that were also affected by the host presence. The results indicate that T. infestans were able to detect the non-photic cycle of their potential hosts, an ability that surely facilitates feeding and hinders predation risk. Understanding triatomines behavior is of fundamental importance to the design of population control methods.     

Developmental rates of two congeneric parasitoids,Encarsia formosa and E. pergandiella (Hymenoptera: Aphelinidae), utilizing different egg provisioning strategies

Endoparasitic Hymenoptera vary in the extent to which they provision their eggs and thus in the degree to which they appear to rely on their hosts for resources during embryonic development. In this study, developmental rates were examined in two congeneric parasitoid species, Encarsia formosa and E. pergandiella, that provision their eggs to different degrees. E. formosa eggs are much larger than E. pergandiella eggs. E. formosa eggs hatch significantly earlier than the eggs of E. pergandiella when deposited in 1st or 4th instar nymphs of a common whitefly host, Bemisia tabaci. Both species hatch earlier in 4th instar nymphs, but the delay in hatching in hosts parasitized as 1st instars is much greater in E. pergandiella. While E. formosa develops more rapidly to the 1st larval instar, E. pergandiella emerge as adults significantly earlier, though smaller, than E. formosa adults regardless of the host instar parasitized. These findings show that the extent of provisioning in the eggs of these wasps does not strictly determine their order of progression through different stages of development.     

Trypanosoma cruzi-infected Rhodnius prolixus endure increased predation facilitating parasite transmission to mammal hosts

Triatomine bugs aggregate with conspecifics inside shelters during daylight hours. At dusk, they leave their refuges searching for hosts on which to blood feed. After finding a host, triatomines face the threat of being killed, because hosts often prey on them. As it is known that many parasites induce the predation of intermediate hosts to promote transmission, and that ingestion of Trypanosoma cruzi-infected bugs represents a very effective means for mammal infection, we hypothesized that trypanosomes induce infected bugs to take increased risk, and, as a consequence, be predated when approaching a host. Therefore, we evaluated whether the predation risk and predation rates endured by Rhodnius prolixus increase when infected with T. cruzi. Assays were performed in square glass arenas offering one central refuge to infected and uninfected 5^th instar nymphs. A caged mouse was introduced in each arena after a three-day acclimation interval to activate sheltered insects and induce them to approach it. As hypothesized, a significantly higher proportion of infected insects was predated when compared with uninfected ones (36% and 19%, respectively). Indeed, T. cruzi-infected bugs took higher risk (Approximation Index = 0.642) when compared with healthy ones (Approximation Index = 0.302) and remained outside the shelters when the host was removed from the arena. Our results show that infection by T. cruzi induces bugs to assume higher risk and endure higher predation rates. We reveal a hitherto unknown trypanosome-vector interaction process that increases infected bug predation, promoting increased rates of robust oral transmission. The significant consequences of the mechanism revealed here make it a fundamental component for the resilient maintenance of sylvatic, peridomestic and domestic cycles.     

Potential population growth and harmful effects on humans from bed bug populations exposed to different feeding regimes

Effects of host availability and feeding period on bed bugs, Cimex lectularius (L.) (Hemiptera: Cimicidae), were measured. Population growth and the potential harmful effect of bed bug populations on human hosts were modelled. Bloodmeal sizes were affected by both feeding length and frequency, with >2‐fold difference between insects fed daily or weekly. Blood consumption increased >2‐fold between bed bugs fed occasionally and often, and 1.5‐fold between occasional and daily feeding. Bed bugs fed more often than once a week, potentially every 2–4 days. Egg production was associated with nutrition, being strongly correlated with blood consumption in the previous week. Bed bug populations can grow under different feeding regimes and are hard to control with <80% mortality. Bed bugs can survive and grow even in locations with a limited blood supply, where bed bug persistence may be important for the continual spread of populations. Persistence in non‐traditional locations and a potential association with human pathogens increase the health risks of bed bugs. Potential blood loss as a result of a bed bug can have serious consequences because uncontrolled populations can reach harmful levels in 3–8 months. The reproduction potential of bed bug populations suggests serious consequences to human health and the need for efficacious control measures.     

RNA interference of NADPH-cytochrome P450 reductase results in reduced insecticide resistance in the bed bug, Cimex lectularius

Background

NADPH-cytochrome P450 reductase (CPR) plays a central role in cytochrome P450 action. The genes coding for P450s are not yet fully identified in the bed bug, Cimex lectularius. Hence, we decided to clone cDNA and knockdown the expression of the gene coding for CPR which is suggested to be required for the function of all P450s to determine whether or not P450s are involved in resistance of bed bugs to insecticides.

Methodology/Principal Findings

The full length Cimex lectularius CPR (ClCPR) cDNA was isolated from a deltamethrin resistant bed bug population (CIN-1) using a combined PCR strategy. Bioinformatics and in silico modeling were employed to identify three conserved binding domains (FMN, FAD, NADP), a FAD binding motif, and the catalytic residues. The critical amino acids involved in FMN, FAD, NADP binding and their putative functions were also analyzed. No signal peptide but a membrane anchor domain with 21 amino acids which facilitates the localization of ClCPR on the endoplasmic reticulum was identified in ClCPR protein. Phylogenetic analysis showed that ClCPR is closer to the CPR from the body louse, Pediculus humanus corporis than to the CPRs from the other insect species studied. The ClCPR gene was ubiquitously expressed in all tissues tested but showed an increase in expression as immature stages develop into adults. We exploited the traumatic insemination mechanism of bed bugs to inject dsRNA and successfully knockdown the expression of the gene coding for ClCPR. Suppression of the ClCPR expression increased susceptibility to deltamethrin in resistant populations but not in the susceptible population of bed bugs.

Conclusions/Significance

These data suggest that P450-mediated metabolic detoxification may serve as one of the resistance mechanisms in bed bugs.     

Amines from vertebrates guide triatomine bugs to resources

Most triatomine bugs (Heteroptera: Reduviidae) are nest-living insects that require vertebrate blood or invertebrate haemolymph to complete their life cycle. Vertebrates accumulate excretory products in or near their nesting sites and we hypothesize that triatomines use emanations from such host wastes when searching for resources. Here we recount how triatomine bugs increase upwind locomotion on a servosphere in response to volatile amine constituents of vertebrate excretions. Fresh chicken faeces is strongly attractive to Rhodnius prolixus nymphs. Ammonia induces attraction and an increase in both speed and total path length by R. prolixus on the servosphere. Whereas ethylamine and dimethylamine attract R. prolixus, Triatoma infestans and Panstrongylus geniculatus, other amine constituents of vertebrate excretions such as isobutylamine and hexylamine induce R. prolixus nymphs to walk faster and for a longer period. These amines are derived from generally occurring metabolites of vertebrates and from gut flora metabolism. We conclude that amines and other products associated with nesting hosts serve as signals for foraging triatomines.     

Biology of Tupiocoris cucurbitaceus (Hemiptera: Miridae), a predator of the greenhouse whitefly Trialeurodes vaporariorum (Hemiptera: Aleyrodidae) in tomato crops in Argentina

The predator Tupiocoris cucurbitaceus is frequently found attacking Trialeurodes vaporariorum in greenhouses without pesticide applications in Argentina. The objective of these studies was to evaluate some biological characteristics of this species fed on three types of diet (whitefly nymphs, Sitotroga cerealella eggs and a mix of both) and on two host plants (tomato and tobacco), under controlled experimental conditions. Preimaginal developmental time for female and male bugs was shorter in the presence of whiteflies than with only moth eggs. Females lived longer when they ate only whitefly nymphs compared to a mixed diet or only moth eggs. The amount of adult descendants was greater when bugs could eat whiteflies, regardless of the presence of S. cerealella. Embryonic development time, male longevity and sex proportion were not affected by the diet or the host plant. Prey consumption was evaluated for three T. cucurbitaceus life history stages (fourth/fifth instar nymphs, female and male adults) on two types of prey (whitefly nymphs and S. cerealella eggs). On tomato, females were more voracious than males and nymphs. On tobacco, adults and nymphs consumed more S. cerealella than T.vaporariorum nymphs, but again, bug females preyed more than males and nymphs. Results demonstrate that T. cucurbitaceus can survive, develop and reproduce normally using both T. vaporariorum and S. cerealella eggs as prey on tobacco or tomato plants. This information can be useful for managing this predator against T. vaporariorum through conservative or augmentative biological control strategies.     

Sex ratios in natural populations ofAphelinus mali (Hym.: Aphelinidae) in relation to host size and host density

In an apple orchard at Armidale, the Northern Tablelands of NSW, population sex ratios ofAphelinus mali (Haldeman), an endoparasitoid of the woolly apple aphid,Eriosoma lanigerum (Hausmann) varied from 0.51 (proportion of males) at low host densities to female-biased at high host densities (proportion of males ranged from 0.35–0.39). This shift in sex ratio seems to be caused by the differences in allocation of sons and daughters to hosts of different sizes. In the fieldA. mali parasitizes all life stages (four nymphal instars and adult) of the woolly aphid upon encountering. According to Hughes'(1979) optimal diet model, such general host acceptance seems to be the best strategy. However, it allows the host nymphs or adults to continue to develop or reproduce until about to mummify (pupate). No mortality was observed when first or second-instar hosts were parasitized in the laboratory. Field collected small mummified hosts yielded male-biased sex ratios whereas large mummified hosts produced mainly females. In the laboratory, progeny from smaller hosts (first to third-instars) produced sex ratios which were not significantly different from 0.5 whereas progeny from larger hosts (third and fourth-instars) produced female-biased sex ratio. During winter (June–August) and early spring (September–October) when the host populations in the orchard were predominantly nymphs, the parasitoid tended to allocate equal resources to male and female offspring. In contrast, at peak population densities in summer and autumn (December–May) when larger hosts were available, the sex ratios were female-biased. The host size ofE. lanigerum andA. mali is, therefore, an important component in the dynamics of host-parasitoid interactions.     

Influence of parasitoid-host density and host availability on the laboratory propagation ofMicroctonus aethiops [Hym.: Braconidae] parasitoid ofHypera postica [Coleop.: Curculionidae]

Laboratory studies of the arrhenotokous braconid wasp,Microctonus aethiops (Nees) were conducted relative to maximizing parasitoid production without adversely affecting sex ratios of the progeny. The influence of hostHypera postica (Gyllenhal) and parasitoid densities on parasitoid productivity was evaluated. Maximum production or a mean 25.8M. aethiops progeny were obtained per female when the host parasitoid ratio was 50∶1. More than one parasitoid per rearing cage resulted in fewer parasitoid progeny per female and increased host mortality. Oviposition significantly increased when female parasitoids were exposed to 25 different weevils every 24 hours. The most intense ovipositional activity usually occurred during the first 48 hours of exposure. However, superparasitism occurred if exposure to 25 hosts continued for more than 24 hours. Continuous and discontinuous oviposition patterns did not affect the numbers of parasitoid progeny produced. However, fewer females were produced when new weevils were provided every 24 hours as in the continuous oviposition group.     

Consequences of mixed species infestation on the searching behavior and parasitism success of a larval parasitoid

When two herbivore pest species are potential hosts of a single parasitoid species, two questions arise. Firstly, which host is preferable for mass rearing in terms of later parasitoid performance, and secondly, how do parasitoids perform in mixed herbivore situations after colony establishment? We tested Hyssopus pallidus, a gregarious parasitoid of two major pests of apple, Cydia (Grapholita) molesta and Cydia pomonella, before and after landing on apples infested by one of the two Cydia species. Pre-alighting host preference was tested in a Y-tube olfactometer setup, and parasitism success in a contact bioassay. To gain information on parasitoid performance throughout the growing season, different fruit growth stages were used. Irrespective of the host they had developed on, the parasitoids showed similar olfactory preferences when given a dual choice between infested and healthy fruits, and they did not discriminate between fruits infested by C. molesta and C. pomonella. Responsiveness was generally high, especially late in the season close to harvest. Both hosts are parasitized regardless of the host the parasitoid female had developed on, and no differences in parasitism rates or number of offspring were noted for the two hosts offered. Results were consistent for all apple growth stages tested. In conclusion, mass rearing of this parasitoid can be carried out on either host, without limiting the future efficacy of the bio-control agent. Similarly, established colonies are expected to develop further on both hosts without any bias in host preference.     

Overwintering Stages of Meloidogyne incognita in Vitis vinifera

The overwintering of Meloidogyne incognita in and around Vitis vinifera cv. French Colombard roots was studied in a naturally infested vineyard at the Kearney Agricultural Center, in a growth chamber, in inoculated vines in microplots at the University of California, Davis, and in a greenhouse. Infected roots were sampled at intervals from onset of vine dormancy until plants accumulated about 800 degree days (DD - base 10 C). Embryogenesis within eggs, classified as less than or more than 16 cells and fully differentiated, and numbers of juveniles (second to fourth stage) and preovipositional and mature (egg-laying) adult stages in roots were determined. All stages were present at the onset of dormancy. Juveniles and immature females were not recovered during the dormant period. Mature females and eggs were always present in roots, although the number of mature females generally decreased with time after onset of dormancy. In contrast, in a greenhouse experiment that accumulated comparable DD without the host plant going through dormancy, the number of mature females increased. After bud break, the number of eggs per female increased and all nematode stages were found in host roots. Eggs in all stages of embryogenesis were observed at all times of sampling, indicating that females overwinter and are capable of laying eggs when conditions improve in the spring and need to be considered in nematode management decisions.     

Utilization of early soybeans for food and reproduction by the tarnished plant bug (Hemiptera: Miridae) in the delta of Mississippi

Commercially produced maturity group (MG) IV soybeans, Glycine max L., were sampled during bloom for tarnished plant bugs, Lygus lineolaris (Palisot de Beauvois), during May and June 1999 (3 fields) and 2001 (18 fields). The adults and nymphs were found primarily in single population peaks in both years, indicating a single new generation was produced during each year. The peak mean numbers of nymphs were 0.61 and 0.84 per drop cloth sample in 1999 and 2001, respectively. Adults peaked at 3.96 (1999) and 3.76 (2001) per sweep net sample (25 sweeps). Tests using laboratory-reared and field-collected tarnished plant bugs resulted in very poor survival of nymphs on 16 different soybean varieties (MG III, one; IV, four; V, nine; VI, two). A large cage (0.06 ha) field test found that the number of nymphs produced on eight soybean varieties after mated adults were released into the cages was lower than could be expected on a suitable host. These results indicated that soybean was a marginal host for tarnished plant bugs. However, the numbers of adults and nymphs found in the commercially produced fields sampled in the study may have been high enough to cause feeding damage to the flowering soybeans. The nature of the damage and its possible economic importance were not determined. Reproduction of tarnished plant bugs in the commercially produced early soybean fields showed that the early soybeans provided tarnished plant bugs with a very abundant host at a time when only wild hosts were previously available.     

Towards a characterization of the locomotor activity rhythm of the desertic species Porcellio olivieri (Crustacea,Isopoda) from Gabes (South of Tunisia)

The locomotor activity rhythm of the isopod, Porcellio olivieri, was investigated in Gannouch site in the south of Tunisia. The rhythm was monitored under constant temperature in individual animals in winter under two simultaneous regimens: the light–dark (LD) cycle and the continuous darkness (DD). Results revealed that whatever regimens, actograms, and mean activity curves showed that specimens of P. olivieri concentrated their activity during the experimental and subjective night. The species exhibited a locomotor rhythm period significantly shorter under LD (T = 23h13 ± 0h44) than DD (τ = 24h28 ± 0h58). However, the locomotor activity rhythm was less stable and the individuals were significantly more active under entraining conditions than constant darkness. The activity pattern of this species will be discussed as an adaptative strategy to respond to environmental conditions.     

Influence of Host Nutrients on the Parasitic Development of Mermis nigrescens (Mermithidae)

Schistocerca gregaria nymphs and adults of both sexes were infected with eggs of Mermis nigrescens. Mermithid larvae grew more slowly in nymphal hosts, and emerging larvae were smaller than those from adult hosts. The longer the larvae remained in the host, the greater their size. Those developing in adult female hosts were longest. Single mermithid larvae that were transferred to a second host continued to grow and were significantly longer at emergence than larvae that developed solely in one host. In adult hosts that were infected with 40-300 M. nigrescens eggs, the percentage of mermithids that became males was strongly dependent on host weight at infective doses of 90 eggs or more. Results are discussed in relation to nutrient stress on the larvae and its importance in developing in vitro culture techniques.     

Competence of Cimex lectularius Bed Bugs for the Transmission of Bartonella quintana,the Agent of Trench Fever

Background Bartonella quintana, the etiologic agent of trench fever and other human diseases, is transmitted by the feces of body lice. Recently, this bacterium has been detected in other arthropod families such as bed bugs, which begs the question of their involvement in B. quintana transmission. Although several infectious pathogens have been reported and are suggested to be transmitted by bed bugs, the evidence regarding their competence as vectors is unclear.ConclusionThe present work demonstrated for the first time that bed bugs can acquire, maintain for more than 2 weeks and release viable B. quintana organisms following a stercorarial shedding. We also observed the vertical transmission of the bacterium to their progeny. Although the biological role of bed bugs in the transmission of B. quintana under natural conditions has yet to be confirmed, the present work highlights the need to reconsider monitoring of these arthropods for the transmission of human pathogens.