Journal: Journal of insect physiology
During larval-pupal transformation, the anterior silk glands (ASGs) of the silkworm Bombyx mori undergo programmed cell death (PCD) triggered by 20-hydroxyecdysone (20E). Under standard in vitro culture conditions (0.3ml of medium with 1μM 20E), ASGs of the fourth-instar larvae do not undergo PCD in response to 20E. Similarly, larvae of the fifth instar do not respond to 20E through day 5 of the instar (V5). However, ASGs of V6 die when challenged by 20E, indicating that the glands might be destined to die before V6 but that a death commitment is not yet present. When we increased the volume of culture medium for one gland from 0.3 to 9ml, V5 ASGs underwent PCD. We examined the response of ASGs to 20E every day by culturing them in 9ml of medium and found that ASGs on and after V2 were fully responsive to 20E. Because pupal commitment is associated with juvenile hormone (JH), the corpora allata (a JH secretory organ) were removed on day 3 of the fourth larval instar (IV3), and their ASGs on V0 were cultured with 20E. Removal of the corpora allata allowed the V0 larval ASGs to respond to 20E with PCD. In contrast, topical application of a JH analogue inhibited the response to 20E when applied on or before V5. We conclude that the acquisition of responsiveness to 20E precedes the loss of JH sensitivity, and that the death commitment in ASGs occurs between V5 and 6.
Several factors threaten the health of honeybees; among them the parasitic mite Varroa destructor and the Deformed Wing Virus play a major role. Recently, the dangerous interplay between the mite and the virus was studied in detail and the transition, triggered by mite feeding, from a benign covert infection to a devastating viral outbreak, characterized by an intense viral replication, associated with some characteristic symptoms, was described. In order to gain insight into the events preceding that crucial transition we carried out standardized lab experiments aiming at studying the effects of parasitization in asymptomatic bees to establish a relationship between such effects and bee mortality. It appears that parasitization alters the capacity of the honeybee to regulate water exchange; this, in turn, has severe effects on bee survival. These results are discussed in light of possible novel strategies aiming at mitigating the impact of the parasite on honeybee health.
The cricket (Gryllus bimaculatus) is a hemimetabolous insect that is emerging as a model organism for the study of neural and molecular mechanisms of behavioral traits. However, research strategies have been limited by a lack of genetic manipulation techniques that target the nervous system of the cricket. The development of a new method for efficient gene delivery into cricket brains, using in vivo electroporation, is described here. Plasmid DNA, which contained an enhanced green fluorescent protein (eGFP) gene, under the control of a Gryllus bimaculatus actin (Gb'-act) promoter, was injected into adult cricket brains. Injection was followed by electroporation at a sufficient voltage. Expression of eGFP was observed within the brain tissue. Localized gene expression, targeted to specific regions of the brain, was also achieved using a combination of local DNA injection and fine arrangement of the electroporation electrodes. Further studies using this technique will lead to a better understanding of the neural and molecular mechanisms that underlie cricket behaviors.
Bombus terrestris colonies go through two major phases: the “pre-competition phase” in which the queen is the sole reproducer and aggression is rare, and the “competition phase” in which workers aggressively compete over reproduction. Conflicts over reproduction are partially regulated by a group of octyl esters that are produced in Dufour’s gland of reproductively subordinate workers and protect them from being aggressed. However, workers possess octyl esters even before overt aggression occurs, raising the question of why produce the ester-signal before it is functionally necessary?. In most insect societies, foragers show reduced aggression and low dominance rank. We hypothesize that ester production in B. terrestris is not only correlated with sterility but also with foraging, signaling cooperative behavior by subordinate workers. Such a signal helps to maintain social organization, reduce the cost of fights between reproductives and helpers, and increase colony productivity, enabling subordinates to gain greater inclusive fitness. We demonstrate that foragers produce larger amounts of esters compared to non-foragers, and that their amounts positively correlate with foraging efforts. We further suggest that task performance, potential fecundity, and aggression are interlinked, and that worker-worker interactions are involved in regulating foraging behavior. B. terrestris, being an intermediate phase between primitive and derived eusocial insects, provides an excellent model for understanding the evolution of early phases of eusociality. Our results, combined with those in primitively eusocial wasps, suggest that at early stages of social evolution, reproduction was regulated by a “primordial division of labor”, that comprised foragers and reproducers, which further evolved to a more complex division of labor, a hallmark of eusociality.
An ability to predict forthcoming changes in environmental conditions and get prepared for them in advance is crucial for the survival and reproduction of organisms living in a seasonally changing environment. We have studied the possible involvement of circadian oscillator(s) in the photoperiodic timer controlling seasonal responses by tracing Drosophila montana females' diapause induction in constant darkness and in a classical Nanda-Hamner experiment. Nearly all females developed ovaries in continuous darkness, which shows the direct development to be their default developmental pathway in the absence of photoperiods. In Nanda-Hamner experiment the females' diapause incidence was close to zero in light:dark cycle 12:4 (photoperiod 16 h) and increased to nearly 100% in 12:8 and 12:12 (photoperiods 20 and 24 h). In longer photoperiods (28-72 h) the females' diapause percentages decreased gradually along with an increase in the length of the dark period, showing no peaks of high diapause incidence in the multiples of 24 h. These findings suggest that the photoperiodic timer of D. montana is based on heavily damping circadian oscillator(s) or that it lacks strong oscillators. Damping of the photoperiodic timer under prolonged nights and constant darkness fits well with our earlier finding that these flies lose their locomotor activity rhythm in constant darkness, and suggests that the mechanisms underlying females' photoperiodic diapause response and their free-running locomotor activity rhythm may be partly based on same oscillators.
Biogenic amines are known to play critical roles in key insect behaviors such as feeding and reproduction. This study documents the effects of reserpine on mating and egg-laying behaviors of the stable fly, Stomoxys calcitrans (L.) (Diptera: Muscidae), which is one of the most significant biting fly pests affecting cattle. Two sperm-staining techniques were adapted successfully to reveal the morphology of stable fly sperm, for the first time, and determine successful mating in females through the assessment of sperm transfer. This approach was also applied to assess sperm transfer by males treated with different doses of reserpine. Mating or sperm transfer did not occur in flies during the first 3 day after emergence. Thereafter, the percentage of females that mated increased with age. Reserpine treatment of males reduced sperm transfer in a dose dependent manner. Older males were more sensitive to reserpine treatment than younger flies. Reserpine treatment of 5 d old females reduced the number of eggs laid, but had no effect on egg hatching rates. Results of immunoreactivity (IR) experiments indicated that serotonin in the neuronal processes innervating male testes was completely depleted by reserpine within five hours after treatment. This effect was transient as the serotonin immunoreactive signal was recovered in 33.3% of the males at 1 d post treatment and in 94.4% of the flies at 3 d post treatment. The results of this study concur with previous findings in other insect species and extend our knowledge of the critical roles biogenic amines play in mating and oviposition behaviors of the stable fly. The work could provide a foundation to further characterize the specific roles of individual biogenic amines and their receptors in stable fly reproduction.
Blood-sucking insects strongly rely on olfactory cues to find their vertebrate hosts. As in other insects with different lifestyles, it has been shown that endogenous and exogenous factors modulate olfactory responses. The triatomine bug Rhodnius prolixus is an important vector of Chagas disease and a classical model for studies of physiology and behavior. In this species, the behavioral response to host-derived odorants is modulated by both the time of the day and the starvation. Here I investigated the peripheral neural mechanisms underlying these modulatory effects. For this, I measured the electroantennogram (EAG) responses of insects towards different concentrations (from 0.5% to 75% vol/vol) of an attractive host-odorant, ammonia. I tested the responses of starved and fed animals during the middle of the day (when insects are inactive and aggregated in refuges) and at the beginning of the night (when insects become active and search for hosts). Regardless of the time of the day and the starvation status, EAG responses systematically increased with odorant concentration, thus accurately reflecting the response of olfactory receptor cells. Interestingly, the EAG responses of starved insects were larger than those of fed insects only during the night, with larger differences (6-7 times) observed at low-middle concentrations. This is study is, to my knowledge, the first reporting modulation of sensory responses at the neural level in triatomines. This modulation, considering that triatomine hosts are mostly diurnal and are also potential predators, has an important adaptive value, ensuring that insects search for hosts only when they are hungry and at appropriate times.
Maternally inherited endosymbiotic bacteria of the genus Wolbachia cause various reproductive alterations in their hosts. Wolbachia induces male-specific death during embryonic and larval stages in the moth Ostrinia scapulalis. To investigate how the density of Wolbachia affects their performance in the host, we attempted to reduce its density using a short, high-temperature treatment of the host at the larval stage. Individuals cured of infection as well as sexual mosaics, which harbor Wolbachia, were obtained by this method in the next generation. The sex of uninfected offspring was exclusively male, similar to that of the offspring of larvae treated with antibiotics. A strong correlation was found between Wolbachia density in female moths and the sex ratio of their progeny. These results suggest that a short, high-temperature treatment at the larval stage reduced the density of Wolbachia in the adult stage, and, hence, inhibited interference with the host’s development in the next generation. Since the direct effects of the heat treatment on Wolbachia were transient, this method may be useful for specifying the critical time for interference by Wolbachia in host development.
While pheromone traps have been effectively used to monitor the recent range expansion of the western bean cutworm (WBC), very little is known about the pheromone mediated reproductive biology of this species. The age at which females initiated calling (the behaviour, associated with the release of the sex pheromone), and the pattern of calling on the first three nights following sexual maturation were determined for virgin females held under four temperature regimes (25:20; 25:15; 20:15; 20:10 °C L:D and 16L:8D photoperiod), and two RH (60 and 80%). Regardless of the rearing conditions the pre-calling period (PCP) was always several days post emergence, supporting the hypothesis that WCB is a migrant species. However, surprisingly the length of the PCP was not directly related to mean temperature but rather to the temperature differential between the photophase and the scotophase. The duration of calling increased with female age, but unlike in other moths was not affected by the abiotic factors tested. The relative insensitivity to temperature and humidity, when compared with many other moth species, may be related to the WBC being a univoltine species with a mid-summer flight period. Consequently, there would not be strong selection pressure for plasticity in calling behavior when compared the case with multivoltine species that experience a wide range of environmental conditions during different seasonal flight periods.
For generalist parasitoids such as those belonging to the Genus Aphidius, the choice of host species can have profound implications for the emerging parasitoid. Host species is known to affect a variety of life history traits. However, the impact of the host on thermal tolerance has never been studied. Physiological thermal tolerance, enabling survival at unfavourable temperatures, is not a fixed trait and may be influenced by a number of external factors including characteristics of the stress, of the individual exposed to the stress, and of the biological and physical environment. As such, the choice of host species is likely to also have implications for the thermal tolerance of the emerging parasitoid. The current study aimed to investigate the effect of cereal aphid host species (Sitobion avenae, Rhopalosiphum padi and Metopolophium dirhodum) on adult thermal tolerance, in addition to sex and size, of the aphid parasitoids Aphidius avenae, Aphidius matricariae and Aphidius rhopalosiphi. Results revealed no effect of host species on the cold tolerance of the emerging parasitoid, as determined by CTmin and Chill Coma, for all parasitoid species. Host species significantly affected the size of the emerging parasitoid for A. rhopalosiphi only, with individuals emerging from R. padi being significantly larger than those emerging from S. avenae, although this did not correspond to a difference in thermal tolerance. Furthermore, a significant difference in the size of male and female parasitoids was observed for A. avenae and A. matricariae, although, once again this did not correspond to a difference in cold tolerance. It is suggested that potential behavioural thermoregulation via host manipulation may act to influence the thermal environment experienced by the wasp and thus wasp thermal tolerance and, in doing so, may negate physiological thermal tolerance or any impact of the aphid host.