Journal of insect physiology最新文献

The spotted-wing fly, Drosophila suzukii, is a world-wide pest insect for which there is increasing interest in its physiological traits including metabolism and thermotolerance. Most studies focus only on survival to different time exposures to extreme temperatures, mainly in female flies. In addition, it has not been tested yet how anesthesia affects these measurements. We analyzed the effects of anesthesia by brief exposures to cold, anoxia by CO₂ or N₂ on three standard thermotolerance assays, as well as the aerobic metabolic rate in both sexes. For heat tolerance we measured CTmax by thermolimit respirometry, and CTmin and chill-coma recovery time for cold tolerance. Aerobic metabolism was calculated by CO₂ production of individual flies in real time by open flow respirometry. Results showed that females have a significantly higher $\dot{V}$CO₂ for inactive (at 25 °C) and maximum metabolic rate than males. This difference is mainly explained by body mass and disappears after mass correction. Males had a more sensitive MR to temperature than females showed by a significantly higher Q₁₀ (2.19 vs. 1.98, for males and females, respectively). We observed a significantly lower CTmin (X² = 4.27, P = 0.03) in females (3.68 ± 0.38 °C) than males (4.56 ± 0.39 °C), although we did not find significant effects of anesthesia. In contrast, anesthesia significantly modifies CTmax for both sexes (F_3,62 = 7.86, P < 0.001) with a decrease of the CTmax in cold-anesthetized flies. Finally, we found a significantly higher CTmax in females (37.87 ± 0.07 °C) than males (37.36 ± 0.09 °C). We conclude that cold anesthesia seems to have detrimental effects on heat tolerance, and females have broader thermotolerance range than males, which could help them to establish in invaded temperate regions with more variable environmental temperatures.

Immune responses to infection result in behavioral changes that affect resource acquisition, such as general starvation and compensatory feeding to offset changes in resource allocation. Mormon crickets aggregate and march in bands containing millions of insects. Some bands are comprised of insects seeking proteins. They are also low in circulating phenoloxidase (PO) and more susceptible to fungal attack, as we have demonstrated in the lab. Here, we ask: Do Mormon crickets elevate PO and consume protein in response to infection by the pathogenic fungus Beauveria bassiana? B. bassiana was applied topically (day 0), and mortality began on day 5. Total protein, PO, and prophenoloxidase (proPO) were assayed in hemolymph on day 1 and 4. On day 1, PO titers were not different between inoculated and control insects, whereas by day 4, PO was greater in the inoculated group. proPO activity was unchanged. Circulating protein declined in inoculated insects relative to controls. As predicted, PO titers were elevated as a result of fungal infection, and hemolymph protein was reduced, but the insects did not compensate behaviorally. Indeed, during the first three days post-infection, infected insects reduced protein consumption while maintaining carbohydrate consumption similar to the controls. Following day 3, a more general reduction in protein and carbohydrate intake was evident in infected insects. Survivorship to infection was associated with the amount of protein consumed and unrelated to carbohydrate consumption. Selective protein deprivation by the host seems counterintuitive, but it might limit growth and toxin production by the invading fungus. Alternatively, the fungus might control the host diet to compromise host immunity to infection. Abrupt changes in allocation resulting from an infection can lead to changes in acquisition that are not always intuitive. Because protein acquisition drives aggression between members of the migratory band, B. bassiana application may reduce cannibalism and slow band movement.

In the present study, we have demonstrated a correlation in time between changes in the amount of apolipophorin III (apoLp-III) in the fat body and hemocytes of Galleria mellonella larvae challenged with Pseudomonas aeruginosa exotoxin A (exoA). An increase in the amount of apoLp-III was detected 1–8 h after the challenge; then, a temporary decrease was observed after 15 h followed by an increase in the level of apoLp-III, however to a different extent. The profile of apoLp-III forms in the hemolymph, hemocytes, and fat body of the exoA-challenged larvae was analyzed using two-dimensional electrophoresis (IEF/SDS-PAGE) and immunoblotting with anti-apoLp-III antibodies. Two apoLp-III forms differing in isoelectric point values estimated at ∼ 6.5 and ∼ 6.1 in the hemolymph and ∼ 6.5 and ∼ 5.9 in the hemocytes as well as one isoform with pI ∼ 6.5 in the fat body with an additional apoLp-III-derived polypeptide with estimated pI ∼ 6.9 were detected in the control insects. The injection of exoA caused a significant decrease in the abundance of both apoLp-III isoforms in the insect hemolymph. In the hemocytes, a decrease in the amount of the pI ∼ 5.9 isoform was detected, while the major apoLp-III isoform (pI ∼ 6.5) remained unchanged. In addition, appearance of an additional apoLp-III-derived polypeptide with an estimated pI ∼ 5.2 was observed. Interestingly, there were no statistically significant differences in the amount of the main isoform in the fat body between the control and exoA-challenged insects, but the polypeptide with pI ∼ 6.9 disappeared completely. It should be noted that the decrease in the amount of apoLp-III and other proteins was especially noticeable at the time points when exoA was detected in the studied tissues.

Parasitoid wasps are haplodiploid, meaning that sperm stored by egg laying females are only used to produce daughters. Thus, the sex ratio of the offspring depends on the availability of sperm after mating. In these insects, males are sensitive to temperature at the pupal stage. This stress leads to subfertility due to a drastic reduction in the number of sperm produced and transferred to females. Experiments were conducted under controlled conditions on the parasitoid wasp Cotesia typhae (Hymenoptera, Braconidae), a natural enemy of the invading pest Sesamia nonagrioides (Lepidoptera, Noctuidae). At 25–27 °C, sperm production was measured for 7 days, and found to reach a plateau at the third day of adult life. It leads to a final amount around 25,000 sperm per male. A male can successfully inseminate at least 10 females, producing predominantly female offspring. Sperm production decreased significantly after 1 day of pupal exposure to heat at 34 or 36 °C and 7 days of cold at 0, 5 or 10 °C. This highlights that both cold and heat are stressful. After mating with one male treated at 10 or 34 °C, females store fewer sperm than the control, and produce fewer daughters. The sex ratio of the offspring is male biased when males experienced temperature stresses during development, like other parasitoid wasps. In the field, C. typhae populations would be affected by heat and cold, at least at the pupal stage. This lowers overwintering risk in case this biological agent was introduced in Europe. This risk is both economical, as companies seek to establish costly continuous production to sell beneficial insects, and ecological as the introduced population would not settle in the ecosystem. Lastly, the transport and storage of this insect of agronomic interest would need to consider temperature variations to ensure successful application.

The renal (Malpighian) tubules of insects play important roles in hemolymph Ca²⁺ regulation. Here we investigated how dietary Ca²⁺ loads from sucrose or blood meals affect the Ca²⁺ content and mRNA expression of Ca²⁺ transporters in Malpighian tubules of adult female mosquitoes. Using the yellow fever mosquito Aedes aegypti we found that feeding females 10% sucrose with elevated Ca²⁺ concentration ad libitum for 6 days led to increased Ca²⁺ content in Malpighian tubules. The increases of Ca²⁺ content correlated with up-regulations of mRNAs encoding intracellular Ca²⁺-ATPases (SERCA and SPCA), a plasma membrane Ca²⁺-ATPase (PMCA), and a K⁺-dependent Na⁺/Ca²⁺ exchanger (NCKX1). We also found that when adult females were fed blood, tubule Ca²⁺ content changed dynamically over the next 72 h in a manner consistent with redistribution of tubule Ca²⁺ stores to other tissues (e.g., ovaries). The changes in tubule Ca²⁺ were correlated with dynamic changes in mRNA abundances of SERCA, SPCA, PMCA, and NCKX1. Our results are the first to demonstrate that Malpighian tubules of adult female mosquitoes have a remarkable capacity to handle high dietary Ca²⁺ loads, most likely through the combination of storing excess Ca²⁺ within intracellular compartments and secreting it into the tubule lumen for excretion. Our results also suggest that the Malpighian tubules play key roles in supplying Ca²⁺ to other tissues during the processing of blood meals.

X-ray micro-CT has been used to study the tracheal system of Pre and Post hibernation Queen wasps (Vespula vulgaris) and their workers. We have compared our findings in wasps with Snodgrass’s description of the tracheal system of the honeybee as characterised by anatomical dissection. Our images, whilst broadly similar, identify the tracheal system as being considerably more complex than previously suggested. One of the 30 wasps imaged had a markedly different, previously undescribed tracheal system. Since completing this study, a large micro-CT study from the American Museum of Natural History (AMNH) has been published. This used different software (Slicer) and analysed 16bit digital data. We have compared our methods with that described in the AMNH publication, adopted their suggested nomenclature and have made recommendations for future studies.

The Proboscis Extension Reflex (PER) paradigm trains honey bees to associate an odor with a sugar reward and is commonly used to assess impacts on associative learning after exposure to pesticides. While the effects of some types of pesticides have been well-investigated, relatively little attention has been focused on fungicides that are applied to flowering crops. We have previously shown that consumption of field-relevant concentrations of the fungicide Pristine® (active ingredients: 25.2% boscalid, 12.8% pyraclostrobin) impairs honey bee performance in an associative learning assay, but the mechanism of its action has not been investigated. We hypothesized that Pristine® interferes with carbohydrate absorption and/or regulation, thereby disrupting the post-ingestive feedback mechanisms necessary for robust learning. To test this hypothesis, we measured hemolymph glucose and trehalose levels at five time points during the ten minutes after bees consumed a sucrose solution. Pristine®-exposed bees had elevated baseline glucose concentrations in the hemolymph relative to control bees. Hemolymph glucose levels rose significantly within five minutes of feeding in control bees, but not in Pristine®-fed bees. These data suggest that the post-ingestive feedback mechanisms necessary for robust learning are disrupted in bees that have consumed this fungicide, providing a plausible mechanistic explanation for its effects on learning performance in the PER assay. Pristine®-exposed bees may have elevated hemolymph glucose levels because the fungicide elicits an inflammatory response. These results provide additional mechanistic understanding of the negative physiological effects of mitotoxic fungicides on this important pollinator.

Mosquito larvae display a stereotyped escape response when they rest attached to the water surface. It consists in detaching from the surface and diving, to return to the surface after a brief time. It has been shown that this response can be evoked several times, by repeatedly presenting a moving shadow. Diving triggered by a potential danger revealed as a simple bioassay for investigating behavioural responses in mosquito larvae, in particular their ability to learn. In the present work, we describe an automated system, based on video-tracking individuals, and extracting quantitative data of their movements. We validated our system, by reinvestigating the habituation response of larvae of Aedes aegypti reared in the laboratory, and providing original data on field-collected larvae of genera Culex and Anopheles. Habituation could be demonstrated to occur in all the species, even though it was not possible to induce dishabituation in Culex and Anopheles mosquitoes. In addition to non-associative learning, we characterised motor activity in the studied species, thanks to the possibility offered by the tracking system to extract multiple variables. The here-described system and algorithms can be easily adapted to multiple experimental situations and variables of interest.

In the present study, we investigated downstream pathways of cyclic adenosine monophosphate (cAMP) signaling (which is related to prothoracicotropic hormone (PTTH)-stimulated ecdysteroidogenesis) in Bombyx mori prothoracic glands (PGs). Results showed that treatment with either dibutyryl cAMP (dbcAMP) or 1-methyl-3-isobutylxanthine (MIX) inhibited phosphorylation of adenosine 5′-monophosphate-activated protein kinase (AMPK) and activated phosphorylation of the translational repressor, 4E-binding protein (4E-BP), a marker of target of rapamycin (TOR) signaling. A chemical activator of AMPK (5-aminoimidazole-4-carboxamide-1-β-d-ribofuranoside, AICAR) increased dbcAMP-inhibited AMPK phosphorylation and blocked dbcAMP-stimulated phosphorylation of 4E-BP, indicating that inhibition of AMPK phosphorylation lies upstream of dbcAMP-stimulated TOR signaling. Treatment of PGs with dbcAMP and MIX also stimulated phosphorylation of a 37-kDa protein, as recognized by a protein kinase C (PKC) substrate antibody, indicating that cAMP activates PKC signaling. Treatment with either LY294002 or AICAR did not affect dbcAMP-stimulated phosphorylation of the PKC-dependent 37-kDa protein, indicating that cAMP-stimulated PKC signaling is not related to phosphoinositide 3-kinase (PI3K) or AMPK. In addition, dbcAMP-stimulated ecdysteroidogenesis in PGs was partially inhibited by pretreatment with either LY294002, AICAR, or calphostin C. From these results, we concluded that AMPK/TOR/4E-BP and PKC pathways are involved in ecdysteroidogenesis of PGs stimulated by cAMP signaling in B. mori.

Environmental plastic pollution has significantly increased in the recent decades, and severely impacts economies, human and biodiversity health. Plastics are made of several chemical additives, including bisphenol and phthalate plasticizers such as bisphenol A (BPA) and Di(2-ethylhexyl)phthalate (DEHP). In some animal species, both BPA and DEHP are known as endocrine disruptor compounds, and can alter physiological and metabolic homeostasis, reproduction, development and/or behavior. To date, the impacts of BPA and DEHP have mainly focused on vertebrates, and to a lesser extent, on aquatic invertebrates. Yet, the few studies which examined the effects of DEHP on terrestrial insects also revealed the impacts this pollutant can have on development, hormone titrations, and metabolic profiles. In particular, it has been hypothesized in the Egyptian cotton leafworm Spodoptera littoralis that the observed metabolic alterations could result from the energetic costs necessary for DEHP detoxification or to the dysregulation of hormonally-controlled enzymatic activities. To get additional insights into the physiological effects of bisphenol and phthalate plasticizers on the moth S. littoralis, larvae were fed with food contaminated by BPA, DEHP, or the mixture of both compounds. Then, activities of four glycolytic enzymes, hexokinase, phosphoglucose isomerase, phosphofructokinase, and pyruvate kinase were measured. BPA and/or DEHP had no effects on the activities of phosphofructokinase and pyruvate kinase. Conversely, BPA-contaminated larvae were characterized by a 1.9-fold increase in phosphoglucose isomerase activity, and BPA + DEHP-fed larvae had highly variable hexokinase activity. Overall, since no disruption of glycolytic enzyme was observed in DEHP-contaminated larvae, our work tended to demonstrate that exposure to bisphenol and DEHP increased the amount of oxidative stress experienced.