Insect Physiology

Insects and vertebrates have multiple major physiological systems, each species having a circulatory system, a metabolic system, and a respiratory system that enable locomotion and survival in stressful environments, among other functions. Broadening our understanding of the physiology of Drosophila melanogaster requires the parsing of interrelationships among such major component physiological systems. By combining electrical pacing and flight exhaustion assays with manipulative conditioning, we have started to unpack the interrelationships between cardiac function, locomotor performance, and other functional characters such as starvation and desiccation resistance. Manipulative sequences incorporating these four physiological characters were applied to five D. melanogaster lab populations that share a common origin from the wild and a common history of experimental evolution. While exposure to starvation or desiccation significantly reduced flight duration, exhaustion due to flight only affected subsequent desiccation resistance. A strong association was found between flight duration and desiccation resistance, providing additional support for the hypothesis that these traits depend on glycogen and water content. However, there was negligible impact on rate of cardiac arrests from exhaustion by flight or exposure to desiccant. Brief periods of starvation significantly lowered the rate of cardiac arrest. These results provide suggestive support for the adverse impact of lipids on Drosophila heart robustness, a parallel result to those of many comparable studies in human cardiology. Overall, this study underscores clear distinctions among the connections between specific physiological responses to stress and specific types of physiological performance.

The responses of (i) groups of crowd-reared mature males of desert locust, Schistocerca gregaria to a choice of two columns of air, one permeated with different concentrations of phenylacetonitrile (PAN), the major component of gregarious-phase male-produced pheromone, and the other untreated, and (ii) individual crowd-reared mature males of the insect to varying concentration gradients of PAN, were studied in two different types of arena. In the choice assay, locusts preferred to be within PAN-permeated air column at low relative doses of the pheromone, but away from PAN at high relative doses. In the second assay, individual locusts were arrested close to PAN source at low PAN concentration gradients, but away from the source at high concentration gradients. The results are consistent with two reported releaser functions of the adult male-released pheromone that are dependent on different sensory thresholds: arrestment and cohesion at lower relative concentrations and male-male homosexual avoidance at higher relative concentrations.

Pre-oral digestion is described as the liquefaction of the solid tissues of the prey by secretions of the predator. It is uncertain if pre-oral digestion means pre-oral dispersion of food or true digestion in the sense of the stepwise bond breaking of food polymers to release monomers to be absorbed. Collagenase is the only salivary proteinase, which activity is significant (10%) in relation to Podisus nigrispinus midgut activities. This suggests that pre-oral digestion in P. nigrispinus consists in prey tissue dispersion. This was confirmed by the finding of prey muscles fibers inside P. nigrispinus midguts. Soluble midgut hydrolases from P. nigrispinus were partially purified by ion-exchange chromatography, followed by gel filtration. Two cathepsin L-like proteinases (CAL1 and CAL2) were isolated with the properties: CAL1 (14.7 kDa, pH optimum (pHo) 5.5, km with carbobenzoxy-Phe-Arg-methylcoumarin, Z-FR-MCA, 32 lM); CAL2 (17 kDa, pHo 5.5, km 11 lM Z-FR-MCA). Only a single molecular species was found for the other enzymes with the following properties are: amylase (43 kDa, pHo 5.5, km 0.1% starch), aminopeptidase (125 kDa, pHo 5.5, km 0.11 mM L-Leucine-p-nitroanilide), a-glucosidase (90 kDa, pHo 5.0, km 5 mM with p-nitrophenyl a-D-glucoside). CAL molecular masses are probably underestimated due to interaction with the column. Taking into account the distribution of hydrolases along P. nigrispinus midguts, carbohydrate digestion takes place mainly at the anterior midgut, whereas protein digestion occurs mostly in middle and posterior midgut, as previously described in seed-sucker and blood-feeder hemipterans.

Els resultats d'aquesta Tesi posen de manifest el paper del cervell en el control del desenvolupament larvari de Sesamia nonagrioides. Així, S. nonagrioides és el primer lepidòpter on s' ha trobat que es pot desenvolupar de larva a adult sense cervell. Les ...

Aedes aegypti is a vector of medically important viruses including those causing Zika, dengue, and chikungunya. During mating, males transfer a number of proteins and other molecules to the female and these components of the male ejaculate are essential in shifting female post-mating behaviors in a number of insect species. Because these molecules are highly variable by species, and female post-mating behavior by species is also varied, behavioral assays testing the function of the ejaculate are necessary before we can develop control strategies targeting the mating system to reduce mosquito populations. Because increased survival in mosquitoes strongly increases vectorial capacity and can influence population sizes and potential risk we tested the effect of mating on female survival. The ejaculate can either promote or reduce female survival, as both have been shown in multiple insect species, yet this effect has not been directly assessed in mosquitoes. We compared survival of females in four treatment groups: mated females, virgin females, and virgin females injected with either an extract from the male reproductive glands or a saline control. Survival, blood feeding frequency, fecundity and cumulative net reproductive rate (R 0 ) were determined after multiple feedings from a human host. Our results confirm that male reproductive gland substances increase female fecundity and blood feeding frequency, resulting in dramatic increases in fitness (R 0 ). We also demonstrate, for the first time, an effect of male reproductive gland extracts alone on female survival, regardless of whether or not the female ingested a vertebrate blood meal. Thus, the effects of MAG extract on survival are not secondary effects from altered blood feeding. Collectively, we demonstrate a direct role for Ae. aegypti malederived molecules on increasing female fitness, reproductive success and, ultimately, transmission potential for vector borne pathogens.

Aedes aegypti is a vector of medically important viruses including those causing Zika, dengue, and chikungunya. During mating, males transfer a number of proteins and other molecules to the female and these components of the male ejaculate are essential in shifting female post-mating behaviors in a number of insect species. Because these molecules are highly variable by species, and female post-mating behavior by species is also varied, behavioral assays testing the function of the ejaculate are necessary before we can develop control strategies targeting the mating system to reduce mosquito populations. Because increased survival in mosquitoes strongly increases vectorial capacity and can influence population sizes and potential risk we tested the effect of mating on female survival. The ejaculate can either promote or reduce female survival, as both have been shown in multiple insect species, yet this effect has not been directly assessed in mosquitoes. We compared survival of females in four treatment groups: mated females, virgin females, and virgin females injected with either an extract from the male reproductive glands or a saline control. Survival, blood feeding frequency, fecundity and cumulative net reproductive rate (R 0 ) were determined after multiple feedings from a human host. Our results confirm that male reproductive gland substances increase female fecundity and blood feeding frequency, resulting in dramatic increases in fitness (R 0 ). We also demonstrate, for the first time, an effect of male reproductive gland extracts alone on female survival, regardless of whether or not the female ingested a vertebrate blood meal. Thus, the effects of MAG extract on survival are not secondary effects from altered blood feeding. Collectively, we demonstrate a direct role for Ae. aegypti malederived molecules on increasing female fitness, reproductive success and, ultimately, transmission potential for vector borne pathogens.

Feeding on the blood of warm-blooded vertebrates is associated to thermal stress in haematophagous arthropods. It has been demonstrated that blood-sucking insects protect their physiological integrity either by synthesising heat-shock proteins or by means of thermoregulatory mechanisms. In this work, we describe the first thermoregulatory mechanism in a tick species,Ornithodoros rostratus. By performing real-time infrared thermography during feeding on mice we found that this acarian eliminates big amounts of fluid (urine) through their coxal glands; this fluid quickly spreads over the cuticular surface and its evaporation cools-down the body of the tick. The spread of the fluid is possible thanks to capillary diffusion through the sculptured exoskeleton ofOrnithodoros. We discuss our findings in the frame of the adaptive strategies to cope with the thermal stress experienced by blood-sucking arthropods at each feeding event warm-blooded hosts.

Transepithelial secretion in Malpighian tubules of the yellow fever mosquito (Aedes aegypti) is mediated by active transport of Na + and K + through principal cells and passive Cl Ϫ transport through the shunt. Permeation through the shunt was assessed by measuring transepithelial halide diffusion potentials in isolated perfused Malpighian tubules, after first inhibiting active transport with dinitrophenol. Diffusion potentials were small under control conditions, revealing Eisenman selectivity sequence I (I Ϫ ϾBr Ϫ ϾCl Ϫ ϾF Ϫ ) which is the halide mobility sequence in free solution. Accordingly, electrical field strengths of the shunt are small, selecting halides for passage on the basis of hydrated size. Leucokinin-VIII (LK-VIII) significantly increased the shunt conductance from 57.1 µS/cm to 250.0 µS/cm. In parallel, the shunt selectivity sequence shifted to Eisenman sequence III (Br Ϫ ϾCl Ϫ ϾI Ϫ ϾF Ϫ ), revealing increased electrical field strengths in the shunt, now capable of selecting small, dehydrated halides for passage. High concentrations of peritubular F Ϫ (142.5 mM) duplicated the effects of LK-VIII on shunt conductance and selectivity, suggesting a role for G-protein. In the presence of LK-VIII (or F Ϫ ), coulombic interactions between the shunt and I Ϫ and F Ϫ may be strong enough to cause binding, thereby blocking the passage of Cl Ϫ . Thus, LK-VIII increases both shunt conductance and selectivity, presumably via G-protein.

Although the utilisation of fungal biological control agents to kill insect pests is desirable, it is known that the outcome of infection may be influenced by a number of criteria, including whether or not the target insect is stressed. In the current work, topical treatment of larvae of the lepidopteran pest, Mamestra brassicae, with conidia of Beauveria bassiana, followed by a heat stress (HS; 37 o C for 1 h) 48 h later, resulted in a similar level of larval survival to that occurring for no heat stress (No-HS), fungus-treated larvae. By contrast, when the HS was applied 24 h after fungal treatment, larval survival was significantly increased, indicating that the HS is protecting the larvae from B. bassiana. Similarly, exposure of larvae to a HS provided protection against Metarhizium brunneum (V275) at 48 h (but not 24 h) after fungal treatment. To elucidate the mechanism(s) that might contribute to HS-induced increases in larval survival against fungal infection, the effects of a HS on key cellular and humoral immune responses and on the level of selected heat shock proteins (HSP) were assessed. When larvae were kept under control (No HS) conditions, there was no significant difference in the haemocyte number per ml of haemolymph over a 24 h period. However, exposure of larvae to a HS, significantly increased the haemocyte density immediately after (t= 0 h) and 4 h after HS compared to the No HS controls, whilst it returned to control levels at t=24 h. In addition, in vitro assays indicated that haemocytes harvested from larvae immediately after (0 h) and 4 h (but not 24 h) after a HS exhibited higher rates of phagocytosis of FITC-labelled B. bassiana conidia compared to haemocytes harvested from non-HS larvae. Interestingly, the HS did not appear to increase anti-fungal activity in larval plasma. Western blot analysis using antibodies which cross react with Drosophila melanogaster HSP, resulted in a relatively strong signal for HSP 70 and HSP 90 from extracts of 50,000 and 100,000 haemocytes, respectively, harvested from No-HS larvae. By contrast, for HSP 60, a lysate derived from 200,000 haemocytes resulted in a relatively weak signal. When larvae were exposed to a HS, the level of all three HSP increased compared to the No HS control 4 h and 16 h after the HS. However, 24 h after treatment, any heat stress-mediated increase in HSP levels was minimal and not consistently detected. Similar results were obtained when HSP 90, 70, and 60 levels were assessed in fat body harvested from heat stressed and non-heat stressed larvae. With regard to HSP 27, no signal was obtained even when a lysate from 200,000 haemocytes or three times the amount of fat body were processed, suggesting that the anti-HSP 27 antibody utilised does not cross-react with the M. brassicae HSP. The results suggest that a HS-mediated increase in haemocyte density and phagocytic activity, together with an upregulation of HSP 90 and 70, may contribute to increasing the survival of M. brassicae larvae treated with B. bassiana and M. brunneum (V275).

The Asian tiger mosquito, Aedes albopictus, is a highly invasive mosquito species that transmits chikungunya and dengue. This species overwinters as diapausing eggs in temperate climates. Early diapause termination may be a beneficial strategy for winter mosquito control; however, a mechanism to terminate the diapause process using chemicals is not known. We tested the hypothesis that a hormonal imbalance caused by the administration of juvenile hormone analog would terminate egg diapause in A. albopictus. We tested the insect growth regulator pyriproxyfen on all developmental stages to identify a susceptible stage for diapause termination. We found that pyriproxyfen treatment of mosquito eggs terminated embryonic diapause. The highest rates of diapause termination were recorded in newly deposited (78.9%) and fully embryonated (74.7%) eggs at 0.1 and 1 ppm, respectively. Hatching was completed earlier in newly deposited eggs (25-30 days) compared to fully embryonated eggs (71-80 days). The combined mortality from premature diapause termination and ovicidal activity was 98.2% in newly deposited and >98.9% in fully embryonated eggs at 1 ppm. The control diapause eggs did not hatch under diapausing conditions. Pyriproxyfen exposure to larvae, pupae and adults did not prevent the females from ovipositing diapausing eggs. There was no effect of pyriproxyfen on diapausing egg embryonic developmental time. We also observed mortality in diapausing eggs laid by females exposed to pyriproxyfen immediately after blood feeding. There was no mortality in eggs laid by females that survived larval and pupal exposures. In conclusion, diapausing eggs were the more susceptible to pyriproxyfen diapause termination compared to other life stages. This is the first report of diapause termination in A. albopictuswith a juvenile hormone analog. We believe our findings will be useful in developing a new control strategy against overwintering mosquito populations.

The experiment determined the effects of varying temperatures and relative humidity on the biology and morphometrics of Caryedon serratus. The temperatures were 30, 35, and 40⁰C and relative humidity differences were 60, 65, and 70%. The results show that eggs hatched in 3-4 days at 30⁰C and 60% RH, 4-5 days at 35⁰C and 65% RH, 6-7 days at 40⁰C and 70% RH respectively. Larval period ranges from 19-23 days at 30 C and 60% RH, 14-18 days at 35 C and 65% RH, 14-18 days at 40 C and 70% RH respectively. The morphometric measurements showed that the average length and breadth of eggs were 0.58 mm and 0.15 mm. The mean length and breadth of larval stages were 0.69, 1.27, 2.52, 3.58 mm and 0.35, 0.86, 1.35, and 2.10 mm respectively. The average length and breadth of the pupa were 4.60 mm and 3.00 mm. The mean length and breadth of adults were 4.10 mm and 3.40 mm. The life cycle of bruchid took 33-40, 14-18, and 16-20 days to complete at temperature degrees of 30, 35, 40⁰C and relative humidity ranges of 60, 65, and 70%. Therefore, this work has recommended that the standard optimum temperature and humidity for the best performance of C. serratus particularly on stored groundnut seeds ranges between 30-35⁰C and 60-65% RH. Farmers and all those involved in the storage of groundnut are admonished to be cautious of this period.

The present investigation on biology under laboratory conditions (25±5 o C and 75% RH) of fall armyworm, Spodoptera frugiperda (J. E. Smith) reared on maize crop based natural diet revealed that its life cycle consisted of four distinct stages i.e., egg, larva, pupa and adult. A single female produced 1000±54.54 eggs throughout her entire life span. The incubation period was 2.60±0.54 days with 96.04±2.24% hatchability of eggs. The larval stage was completed by passing through six instars. The average stadial periods for first, second, third, fourth, fifth and sixth instar larvae were 2.40±0.49, 2.50±0.50, 2.20±0.40, 2.90±0.83, 3.00±0.77 and 4.20±0.75 days, respectively. The total larval duration varied from 13 to 22 days, with a mean duration of 17.90±2.76 days. The average pupal period was 11.10±1.45 days. Female moths had a pre-oviposition period of 4.00±0.89 days that included a sexual maturity period. The oviposition and post-oviposition period recorded was 3.00±0.89 and 5.00±0.63 days, respectively. Longevity of males and females were recorded at 8.00±0.89 and 10.80±1.17 days, respectively. The mean total life cycle of males and females were observed to be 37.60±3.67 and 39.40±4.41 days with a range of 34-42 and 35-46 days, respectively.

Many flies have specially evolved feeding mechanisms to imbibe liquids of specific viscosities. Observations of feeding on atypical liquids are notable because of their rarity. We report the first record of intrusive fluid feeding on vertebrate carrion by Philoliche rondani.

A system of markers has been devised to track the development of 3rd and 4th instar/pharate adult greenhouse whiteflies. Instars were identified based on measurements of body width and body length. Depending upon the host plant, the product of the two measurements was exceptionally useful in distinguishing between instars. Body depth was used to divide the 3rd instar into eight stages and body depth and color and appearance of the developing adult eye were used to divide the 4th instar/pharate adult into nine stages. Under conditions of L:D 16:8 and a temperature of 26±2°C, the body depth of 3rd instars reared on greenbean increased from 0.025 (stage 1) to 0.2 mm (stage 8) and the instar duration was approximately 3 days. The body depth of 4th instars increased from approximately 0.1±0.02 (Stage 1) to 0.3±0.03 mm (Stage 5) and then remained constant or decreased slightly during adult development. Ecdysteroid titers peaked at approximately 120 fg/µg protein during Stages 3 through 6 of the 4th instar. Based on an external examination of developing 4th instars and the fluctuations in ecdysteroid titer, it appears that adult development is initiated in Stage 4 or 5 4th instars. Results from histological studies support this view. In Stage 4 nymphs, a subtle change was observed in the corneagenous cells of the eye. However, most Stage 4 4th instars possessed wing development characteristic of earlier, immature stages. In all Stage 5 insects, wing development had been initiated and the corneagenous cells had become quite distinct. In Stage 6 whiteflies, the wing buds were deeply folded and by Stage 7, spines were observed on the new cuticle, indicating that the adult cuticle was well-formed by this stage. Our study is the first to investigate the timing and regulation of the molt, to monitor ecdysteroid titers in precisely staged 4th instar whiteflies and to examine the internal anatomical changes associated with metamorphosis in these tiny homopteran insects. Published by Elsevier Science Ltd.

Temperature and dehydration stress are two major co-occurring environmental stressors threatening the physiology, biochemistry, and ecology of insects. As such, understanding adaptive responses to desiccation stress is critical for predicting climate change impacts, particularly its influence on insect invasions. Here, we assessed water balance and desiccation resistance of the invasive Tuta absoluta (Meyrick, 1917) (Lepidoptera: Gelechiidae), and infer how eco-physiology shapes its niche. We measured basal body water and lipid content, water loss rates (WLRs), and desiccation resistance in larvae (second to fourth instars) and adults. Body -water, -lipid, and WLRs significantly varied across life stages. Second instars recorded the lowest while fourth instars exhibited the highest body water and lipid content. Adult body water and lipid content were higher than second and third instars and lower than fourth instars while proportion of body water and lipid contents were highest in a...

In nature, insects concurrently face multiple environmental stressors, a scenario likely increasing with climate change. Integrated stress resistance (ISR) thus often improves fitness and could drive invasiveness, but how physiological mechanisms influence invasion has lacked examination. Here, we investigated cross‐tolerance to abiotic stress factors which may influence range limits in the South American tomato pinworm—a global invader that is an ecologically and socially damaging crop pest. Specifically, we tested the effects of prior rapid cold‐ and heat‐hardening (RCH and RHH), fasting, and desiccation on cold and heat tolerance traits, as well as starvation and desiccation survivability between T. absoluta life stages. Acclimation effects on critical thermal minima (CTmin) and maxima (CTmax) were inconsistent, showing significantly deleterious effects of RCH on adult CTmax and CTmin and, conversely, beneficial acclimation effects of RCH on larval CTmin. While no beneficial effe...

South American tomato pinworm, Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) is a devastating invasive global insect pest of tomato, Solanum lycopersicum (Solanaceae). In nature, pests face multiple overlapping environmental stressors, which may significantly influence survival. To cope with rapidly changing environments, insects often employ a suite of mechanisms at both acute and chronic time-scales, thereby improving fitness at sub-optimal thermal environments. For T. absoluta, physiological responses to transient thermal variability remain under explored. Moreso, environmental effects and physiological responses may differ across insect life stages and this can have implications for population dynamics. Against this background, we investigated short and long term plastic responses to temperature of T. absoluta larvae (4 th instar) and adults (24-48 h old) field populations. We measured traits of temperature tolerance vis critical thermal limits [critical thermal minima (CTmin) and maxima (CTmax)], heat knockdown time (HKDT), chill coma recovery time (CCRT) and supercooling points (SCP). Our results showed that at the larval stage, Rapid Cold Hardening (RCH) significantly improved CTmin and HKDT but impaired SCP and CCRT. Heat hardening in larvae impaired CTmin, CCRT, SCP, CTmax but not HKDT. In adults, both heat and cold hardening generally impaired CTmin and CTmax, but had no effects on HKDT, SCP and CCRT. Low temperature acclimation significantly improved CTmin and HKDT while marginally compromising CCRT and CTmax, whereas high temperature acclimation had no significant effects on any traits except for HKDT in larvae. Similarly, low and high temperature acclimation had no effects on CTmin, SCPs and CTmax, while high temperature acclimation significantly compromised adult CCRT. Our results show that larvae are more thermally plastic than adults and can shift their thermal tolerance in short and long timescales. Larval plasticity advantage over adults reported here suggest asymmetrical ecological role of the larva relative to adults in facilitating T. absoluta invasion.

A number of monoclonal antibodies have been raised against the haemocytes (blood cells) of two insect species: namely, the wax moth, Galleria mellonella and a species of cockroach, Blaberus discoidalis. In the case of the wax moth, two groups of monoclonal antibodies were raised. One group, designated GM1 to GM28, identified only one type of blood cell, namely the gran-ular cell. All 28 monoclonal antibodies recognised a 90 kDa protein as shown by western blotting. The second group of monoclonal antibodies, designated GallPG1 to GallPG20, recognised all the haemocyte types of G. mel -lonella, including the granular cells, prohaemocytes, plasmatocytes, spherule cells and oenocytoids. These latter monoclonal antibodies all identified a protein of 35 kDa when tested by western blotting. All monoclonal antibodies against G. mellonella haemocytes were highly specific for wax moth haemocytes and did not cross-react with any other lepidopteran, orthopteran or dic-tyopteran haemocytes teste...

Juvenile hormone III (JH) is synthesized by the corpora allata (CA) and plays a key role in mosquito development and reproduction. A decrease in JH titer during the last instar larvae allows pupation and metamorphosis to proceed. As the anti-metamorphic role of JH comes to an end, the CA of the late pupa once again synthesizes JH, which plays an essential role in orchestrating reproductive maturation. In spite of the importance of Aedes aegypti as a vector, a detailed study of the changes of JH hemolymph titers during the gonotrophic cycle has never been performed. In the present studies, using a High Performance Liquid Chromatography coupled to a Fluorescent Detector (HPLC-FD) method, we measured changes in JH levels in the hemolymph of female mosquitoes during the pupal and adult stages. Our results revealed tightly concomitant changes in JH biosynthesis and JH hemolymph titers during the gonotrophic cycle of female mosquito. Feeding high sugar diets resulted in an increase of JH titers, and mating also modified JH titers in hemolymph. In addition these studies confirmed that JH titer in mosquitoes is fundamentally determined by the rate of biosynthesis in the CA.

Juvenile hormone (JH) regulates reproductive maturation in insects; therefore interruption of JH biosynthesis has been considered as a strategy for the development of target-specific insecticides. The corpora allata (CA) from mosquitoes is highly specialized to supply variable levels of JH, which are linked to ovarian developmental stages and influenced by nutritional signals. However, very little is known about how changes in JH synthesis relate to reproductive physiology and how JH synthesis regulation is translated into changes in the CA machinery. With the advent of new methods that facilitate the analysis of transcripts, enzymes and metabolites in the minuscule CA, we were able to provide comprehensive descriptions of the mevalonic (MVA) and JH synthesis pathways by integrating information on changes in the basic components of those pathways. Our results revealed remarkable dynamic changes in JH synthesis and exposed part of a complex mechanism that regulates CA activity. Principal component (PC) analyses validated that both pathways (MVAP and JH-branch) are transcriptionally co-regulated as a single unit, and catalytic activities for the enzymes of the MVAP and JH-branch also changed in a coordinate fashion. Metabolite studies showed that global fluctuations in the intermediate pool sizes in the MVAP and JH-branch were often inversely related. PC analyses suggest that in female mosquitoes, there are at least 4 developmental switches that alter JH synthesis by modulating the flux at distinctive points in both pathways.

The effect of male accessory gland substances on female reproductive physiology has been previously described as "activating" egg development. However, no mechanism has been described that can explain how male mosquitoes are able to influence egg development in female mosquitoes. To investigate how male mosquitoes are able to influence ovarian physiology and reproductive output we explored three main questions: 1) Do mating and male accessory gland substances affect ovarian physiology and alter markers of oocyte quality during the previtellogenic resting stage? 2) Does the male accessory gland contain JH III and is JH III transferred to the female during copulation? 3) Finally, does the nutritional history of the male affect the amount of JH III transferred to the female and alter reproductive output? By answering these questions it is clear that male mosquitoes are able to alter the female's resource allocation priorities towards reproduction by transferring JH III during copulation; reducing the rate of previtellogenic resorption and increasing the amount of stored ovarian lipids. These changes improve an individual follicle's likelihood of development after a blood meal. In addition, males maintained under better nutritional conditions make and transfer more JH III, prevent more follicular resorption and realize higher fecundities than other males. Together these results illustrate one mechanism behind the "activating" effect of mating described as well as the role sugar feeding plays in male mosquitoes.

Juvenile hormone (JH) levels must be modulated to permit the normal progress of development and reproductive maturation in mosquitoes. JH is part of a transduction system that assesses nutritional information and controls reproduction in mosquitoes. Adult female Aedes aegypti show nutritionally-dependent dynamic changes in corpora allata (CA) JH biosynthetic activities. A coordinated expression of most JH biosynthetic enzymes has been described in female pupae and adult mosquitoes; increases or decreases in transcript levels for all the enzymes were concurrent with increases or decreases in JH synthesis; suggesting that transcriptional changes are at least partially responsible for the dynamic changes of JH biosynthesis. The goal of the present study is to identify signaling network components responsible for the nutritionaldependent changes of JH synthesis in the CA of mosquitoes. The insulin/TOR signaling network plays a central role in the transduction of nutritional signals that regulate cell growth and metabolism in insects. These pathways have also been suggested as a link between nutritional signals and JH synthesis regulation in the CA of cockroaches and flies. We used a combination of in vitro studies and in vivo genetic knockdown experiments to explore nutritional signaling pathways in the CA. Our results suggest that the insulin/TOR pathway plays a role in the transduction of the nutritional information that regulates JH synthesis in mosquitoes. Transcriptional regulation of the genes encoding JH biosynthetic enzymes is at least partially responsible for these nutritionally modulated changes of JH biosynthesis.

Nineteen canonical and two Knirps-like family nuclear receptors (NRs) were identified in the genome of Tribolium castaneum. The current study was conducted to determine the function of these NRs in regulation of female reproduction and embryogenesis. RNA interference (RNAi)aided knock-down in the expression of genes coding for 21 NRs showed that seven NRs E75, hormone receptor 3 (HR3), ecdysone receptor (EcR), ultraspiracle (USP), seven-up (SVP), FTZ transcription factor 1 (FTZ-F1) and hormone receptor 4 (HR4) are required for successful vitellogenesis and oogenesis. Knocking down the expression of genes coding for these seven NRs affected egg production by reducing the levels of vitellogenin mRNAs as well as by affecting the oocyte maturation. Expression of seven additional NRs hormone receptor 96 (HR96), hormone receptor 51 (HR51), hormone receptor 38 (HR38), hormone receptor 39 (HR39), Tailless (Tll), Dissatisfaction (Dsf) and Knirps-like is required for successful embryogenesis. The knock-down in the expression of genes coding for three other NRs (E78, hepatocyte nuclear factor 4, HNF4 and Eagle) partially blocked embryogenesis. This study showed that at least 17 out of the 21 NRs identified in T. castaneum play key roles in female reproduction and embryogenesis.

In holometabolous insects, extensive reorganisation of tissues and cells occurs at the pupal stage. The remodelling of the external exoskeleton and internal organs that intervenes during metamorphosis has been traditionally studied in many insect species based on histological or ultrastructural methods. This study demonstrates the use of synchrotron X-ray phase-contrast micro-computed tomography as a powerful, non-destructive tool for in situ morphological observation of anatomical structures at the pupal stage in two Tenebrionid beetles, i.e. Tribolium castaneum and Tenebrio molitor, known as important pests, as well as emerging and promising models in experimental biology. Virtual sections and three-dimensional reconstructions were performed on both males and females at early, intermediate, and late pupal stage. The dataset allowed us to observe the remodelling of the gut and nervous system as well as the shaping of the female and male reproductive system at different pupal ages in both mealworm and red flour beetles. Moreover, we observed that the timing and duration pattern of organ development varied between the species analysed, likely related to the species-specific adaptations of the pre-imaginal stages to environmental conditions, which ultimately affect their life cycle. This research provides new knowledge on the morphological modifications that occur during the pupal stage of holometabolous insects and provides a baseline set of information on beetle metamorphosis that may support future research in forensics, physiology, and ecology as well as an image atlas for educational purposes.

It is long established that queens of social insects, including termites, maintain their reproductive dominance with queen primer pheromones (QPPs). Yet, the QPP chemistry has only been elucidated in a single species of lower termites. By contrast, the most diversified termite family Termitidae (higher termites), comprising over 70% of termite species, has so far resisted all attempts at QPP identification. Here, we show that the queen- and egg-specific sesquiterpene (3R,6E)-nerolidol acts as the QPP in the higher termite Embiratermes neotenicus. This species has a polygynous breeding system, in which the primary queen is replaced by multiple neotenic queens of parthenogenetic origin. We demonstrate that (3R,6E)-nerolidol suppresses the development of these parthenogenetic queens and thus mimics the presence of mature queen(s). It acts as an airborne signal and may be used to optimize the number of queens, thus being the key regulatory element in the special breeding system of E. ne...

During in vitro incubations, the nematobacterial complex Steinernema carpocapsae-Xenorhabdus nematophilus produces different factors having toxic activities in vitro towards haemocytes, the insect cells responsible for cellular immune defense reactions. Among others, two effects were evident on haemocyte monolayers; one of them was a cytotoxic activity while the other was an unsticking effect. The factors responsible for cytotoxic activity and unsticking effect, were separated from each other by a single chromatography on anion exchange column. These two effects on haemocytes were lost after heat treatment at 57°C for 1 h and 45°C for 30 min, respectively. Both factors were recovered after dialysis in a 10 4 Da cut off membrane. The cytotoxic activity was susceptible to proteases. Cytotoxic and unsticking factors did not show any lipase or lecithinase activity but the unsticking factor had protease activity. Lipopolysaccharides, purified from the bacteria harvested after incubation of the complex, did not have cytotoxic or unsticking effect on the insect cells in vitro.

The vitellogenins of Hyalophora cecropia and Blattella germanica have similar Stokes' radii (69 and 75,&), sedimentation coefficients (15.9 and 16.8s) and isoelectric points (pH 5.7 and 5.0), and share similar amino acid compositions with vitellogenins of other animals. The two vitellogenins show no immunological crossreaction. Blattella oocytes take up their own vitellogenin in vivo at a rapid rate and that of Hyalophora at a low rate comparable to that of a non-vitellogenic protein. Hyalophora oocytes take up their own vitellogenin rapidly in vitro and Blattella Vitellogenin only at a negligible rate. Molecular weights, shapes and charges of the vitellogenins are similar to non-vitellogenins and form no basis for selective uptake.

Inorganic polyphosphates (PolyP) are widespread molecules that have been shown to play a role in metal detoxification and heavy-metal tolerance. In the present report, we investigated the functional role of spherites as PolyP-metal binding stores in epithelial cells of the midgut of Anticarsia gemmatalis, a lepidopteran pest of soybean. PolyP stores were detected by DAPI staining and indirect immunohistochemistry as vesicles distributed in columnar cells and around goblet cell cavities. These PolyP vesicles were identified as spherites by their elemental profile in cell lysates that were partially modulated by P-or V-ATPases. PolyP levels along the midgut were detected using a recombinant exopolyphosphatase assay. When copper was added in the diet of larva, copper detection in spherites by X-ray microanalysis correlated with an increase in the relative phosphorous X-ray signal and with an increase in PolyP levels in epithelia cell lysate. Transmission electron microscopy of chemically fixed or cryofixed and freeze substituted tissues confirmed a preferential localization of spherites around the goblet cell cavity. Taken together, these results suggest that spherites store high levels of PolyP that are modulated during metal uptake and detoxification. The similarity between PolyP granules and spherites herein described also suggest that PolyP is one of the main phosphorous source of spherites found in different biological models. This suggests physiological roles played by spherites in the midgut of arthropods and mechanisms involved in heavy metal resistance among different insect genera.

Several lepidopteran pests of cotton have cadherin-based resistance to the Bacillus thuringiensis (Bt) toxin Cry1Ac. Cadherins are transmembrane proteins that mediate cell-cell adhesion and tissue morphogenesis, suggesting that fitness costs associated with cadherin mutations may be present in many aspects of life history. To evaluate whether cadherin-based resistance is associated with fitness costs reducing male paternity in Helicoverpa armigera, we examined the effects of a major cadherin resistance allele on sperm competition within and between male ejaculates. When homozygous resistant and susceptible males competed for fertilization of a homozygous resistant or susceptible female, fertilization success was high in males with a different cadherin genotype than females, and low in males with the same cadherin genotype as females. Single matings between heterozygous males and susceptible females produced offspring within typical Mendelian ratios. Heterozygous males mated to resis...

The Formosan subterranean termite, Coptotermes formosanus, with its huge colonies, is a major urban pest in several southern states and Hawaii as well as in South Asia. Because of their cryptic nature (underground habitat) and very long life cycle, not much is known about molting in termite workers. In C. formosanus, the workers stop foraging and lose their gut fauna, respectively, approximately 10 and 5 days prior to ecdysis. In any given colony an average of 1.01% (range 0.6-1.8) of the workers were found to molt each day under laboratory conditions. Workers destined to molt become sluggish and their head capsules develop a mottled texture one day prior to ecdysis. Ecdysis was generally accomplished with the assistance of other workers, which also fed on the exuviae. Immediately after molting worker mandibles were light pink in color and became fully melanized approximately two days later. Gut fauna were acquired on the fourth day after molting. Flagellates were transferred as small encysted cells from other workers through proctodeal feeding. Juvenile hormone III titer ranged between 30-41 pg/mg bodyweight in all stages except in workers sampled 6 days prior to ecdysis. In these workers the titer was 80.5 pg/mg. The high juvenile hormones (JH) titer may also be involved in causing defaunation. Ecdysteroid titer increased from 2.1 pg/mg in non-molting workers to 359.5 and 332.4 pg/mg one and two days following defaunation, respectively. There was a second smaller peak two days prior to ecdysis.

In heterotherm insects, endothermy implies a high energy cost due to the generation and regulation of body temperature during different activities such as flight, food location, fighting and even walking. We studied the thermoregulation process and the cost of the thermoregulation strategies in two sympatric dung beetles, Sulcophanaeus batesi and S. imperator under heat and cold stress conditions. We used a set of physiological variables to exemplify the capacity of thermolimit respirometry combined with infrared thermography to derive relevant variables capable of describing different thermoregulation syndromes. Habitat use and thermal niche differed notably between S. batesi and S. imperator, reflecting their contrasted thermal requirements. In S. imperator, thermal specialization for high temperatures was observed, being active mainly during the warmer period of the day. On the other hand, thermal adaptation in S. batesi allows its preference for cold exhibiting a morning activity periods, avoiding higher temperatures. The thermophilic strategy used by Sulcophanaeus imperator minimized the energy expenditure produced during the cooling of the body by respiration without thereby endangering higher thermal limits. In this case, S. batesi, the species with a preference for the coldest environments, presented the lowest thermal limits, although the energy cost needed to stay active during cooling was significantly lower than that in S. imperator. Sulcophanaeus imperator and S. batesi showed evident 'economizing' strategies associated with hot and cold environmental conditions, respectively. In contrast, if both species experience a deviation from their thermal optimum, a decrement in their performance could be produced.

In heterotherm insects, endothermy implies a high energy cost due to the generation and regulation of body temperature during different activities such as flight, food location, fighting and even walking. We studied the thermoregulation process and the cost of the thermoregulation strategies in two sympatric dung beetles, Sulcophanaeus batesi and S. imperator under heat and cold stress conditions. We used a set of physiological variables to exemplify the capacity of thermolimit respirometry combined with infrared thermography to derive relevant variables capable of describing different thermoregulation syndromes. Habitat use and thermal niche differed notably between S. batesi and S. imperator, reflecting their contrasted thermal requirements. In S. imperator, thermal specialization for high temperatures was observed, being active mainly during the warmer period of the day. On the other hand, thermal adaptation in S. batesi allows its preference for cold exhibiting a morning activity periods, avoiding higher temperatures. The thermophilic strategy used by Sulcophanaeus imperator minimized the energy expenditure produced during the cooling of the body by respiration without thereby endangering higher thermal limits. In this case, S. batesi, the species with a preference for the coldest environments, presented the lowest thermal limits, although the energy cost needed to stay active during cooling was significantly lower than that in S. imperator. Sulcophanaeus imperator and S. batesi showed evident 'economizing' strategies associated with hot and cold environmental conditions, respectively. In contrast, if both species experience a deviation from their thermal optimum, a decrement in their performance could be produced.

E)-β-farnesene (EBF) is the common active component of aphid alarm pheromone. Either or both of two orthologs of ordorant-binding proteins (OBPs), OBP3 and OBP7, recently reported in aphids, may be involved in EBF perception. The aim of this study was to investigate the respondence of the aphid Rhopalosiphum padi to its intraspecific alarm pheromone and which OBP is responsible for that response. We tested the olfactory response of the aphid R. padi to EBF and freshly crushed aphids. Then, we extracted the volatiles from crushed aphids using solid phase microextraction (SPME) for analysis with GC×GC-TOF/MS. We also cloned two OBPs cDNAs in R. padi (RpadOBP3 and RpadOBP7) and expressed them in competent Escherichia coli cells. Both recombinant proteins, RpadOBP3 and RpadOBP7, bound EBF well, with RpadOBP7 having specifically stronger affinity for EBF than for other volatiles. Based on the crystal structure of the OBPs with high identity, we performed homology modeling and analyzed the interactions between RpadOBPs and EBF. In conclusion, R. padi was repelled by both EBF and crushed aphids. EBF was identified as the only volatile that acted as the alarm pheromone. Our results indicated that OBP7 is a potential molecular target to control wheat aphids by disturbing their behaviors to the alarm pheromone.

The extent to which environmental factors influence the ability of Anopheles mosquitoes to transmit malaria parasites remains poorly explored. Environmental variation, such as change in ambient temperature, will not necessarily influence the rates of host and parasite processes equivalently, potentially resulting in complex effects on infection outcomes. As proof of principle, we used Anopheles stephensi and the rodent malaria parasite, Plasmodium yoelii, to examine the effects of a range of constant temperatures on one aspect of host defense (detected as alterations in expression of nitric oxide synthase gene - NOS) to parasite infection. We experimentally boosted mosquito midgut immunity to infection through dietary supplementation with the essential amino acid l-Arginine (l-Arg), which increases midgut nitric oxide (NO) levels by infection-induced NOS catalysis in A. stephensi. At intermediate temperatures, supplementation reduced oocyst prevalence, oocyst intensity, and sporozoi...

RNA interference (RNAi) is a sequence-specific gene silencing mechanism that holds great promise for effective management of agricultural pests. Previous studies have shown that the efficacy of RNAi varies among different insect species, which limits its wide spread application in the field of crop protection. In this study, we identified and characterized six core RNAi pathway genes including OfDicer1, OfDicer2, OfR2D2, OfAgo1, OfAgo2, and OfAgo3 from the transcriptomic database of the Asian corn borer (Ostrinia furnacalis). Domain analysis showed that the six deduced proteins contained the necessary functional domains. Insect developmental stage- and tissue-specific expression analysis showed that five genes were expressed in all the stages and tissues examined except OfAgo3, which showed low expression in larvae, and high expression in pupae and adults and in the midgut. RT-qPCR was performed to examine the response of these six genes to exogenous double-stranded RNA (dsRNA). Int...

Small interfering RNAs (siRNAs) are non-coding RNAs with a length of 21~23 nucleotides (nt) and present in almost all eukaryotes. The formation of siRNA is a highly conserved post-transcriptional gene-silencing mechanism mediated by key proteins, including Dicer2, Argonaute2 (Ago2) and R2D2. R2D2 has been identified as a double-stranded RNA (dsRNA)-binding protein and reported as an integral component of the siRNA pathway in Drosophila. However, the involvement of R2D2 in the siRNA pathway of Locusta migratoria is still unknown. In the present study, we identified an LmR2D2 gene from the transcriptome of L. migratoria. It consists of a 954-bp open reading frame that encodes a protein of 318 amino acid residues. Further sequence analysis revealed that LmR2D2 possesses two tandem dsRNA-binding domains (dsRBD) at the N-terminus. Analysis of the developmental expression profile of LmR2D2 indicated that its transcript level was stable in third-instar nymphs of L. migratoria, whereas the ...

Western bean cutworm, Striacosta albicosta (Smith) (Lepidoptera: Noctuidae), is a pest of maize and dry beans that has recently undergone a northeastern range expansion in North America. In order to assess the cold tolerance of S. albicosta, we determined the supercooling point of lab and field collected late instar larvae and pre-pupae. Individuals were attached to fine contact thermocouples and cooled at 1°C per minute to detect heat released due to freezing of body fluids. Mean supercooling points decreased as larvae developed into later life stages. Pre-pupa collected in late fall had a mean supercooling point of -12.63°C. This research is the first documentation of cold tolerance measures for S. albicosta and will aid in designing future cold tolerance experiments and predicting S. albicosta population densities based on winter temperatures.

The mounting sex pheromone of Dermacentor variabilis, the American dog tick, is cholesteryl oleate. This steryl ester, when present in amounts from about I to 0.1 female equivalents elicited male mounting responses that were indistinguishable from the natural controls. Other steryl esters present on the tick's body surface also elicited responses, but at significantly lower intensity than cholesteryl oleate. Long regarded as a waste product of vertebrate blood feeding, this material is secreted by sexually mature females during feeding and accumulates on the surface of the body cuticle. Mate-seeking males, attracted to the females by the presence of 2,bdichlorophenol, recognize and mount the feeding females. The accumulation of cholesteryl oleate on the female's body surface during blood feeding is an indication of successful parasitism and acts as a signal of female mating readiness. Compounds of unknown composition that co-chromatographed with steryl esters were also found in fed females of 4 other ixodid tick species. Bioassays performed with crude extracts of these different species suggest that mounting sex pheromones are widespread in the Ixodidae and may also use the same or similar steryl esters, either alone or in mixtures, as in D. variabilis.

Feeding behaviors and biomechanics of female Varroa destructor mites are revealed from AC-DC electropenetrography (EPG) recordings of mites feeding from Apis mellifera honey bee pupae and histology of mite internal ingestion apparatus. EPG signals characteristic of arthropod suction feeding (ingestion) were identified for mites that fed on pupae during overnight recordings. Ingestion by these mites was confirmed afterwards by observing internally fluorescent microbeads previously injected into their hosts. Micrographs of internal ingestion apparatus illustrate the connection between a gnathosomal tube and a pharyngeal lumen, which is surrounded by alternating dilator and constrictor muscles. Inspection of EPG signals showed the muscularized mite pharyngeal pump operates at a mean repetition rate of 4.5 cycles/s to ingest host fluids. Separate feeding events observed for mites numbered between 23 and 33 over approximately 16 h of recording, with each event lasting ~10 s. Feeding events were each separated by ~2 min. Consecutive feeding events separated by either locomotion or prolonged periods of quiescence were grouped into feeding bouts, which ranged in number from one to six. Statistical analyses of EPG data revealed that feeding events were prolonged for mites having lower pharyngeal pump frequencies, and mites having prolonged feeding events went unfed for significantly more time between feeding events. These results suggest that mites may adjust behaviors to meet limitations of their feeding apparatus to acquire similar amounts of food. Data reported here help to provide a more robust view of Varroa mite feeding than those previously reported and are both reminiscent of, as well as distinct from, some other acarines and fluid-feeding insects.

SummaryAllatostatins with a typical C‐terminal sequence YXFGL‐NH2 are insect neuropeptides with inhibitory properties upon Juvenile Hormone production in the corpora allata, vitellogenin release by the fat body, and gut and dorsal vessel motility. All these biological effects are rapidly reversible, suggesting the occurrence of effective mechanisms for inactivation of the peptides. We have studied the degradation of DRLYSFGL‐NH2 (BLAST‐2), one of the allatostatins of Blattella germanica, in the internal milieu of adult females of this cockroach. The experimental approach combined the use of the radioiodinated derivative [125I‐Tyr4]BLAST‐2, microdialysis techniques and HPLC analysis with a radioisotope detector. Under these experimental conditions, the half‐life of BLAST‐2 in the internal milieu of the adult female of B. germanica was between 3 and 6 min. Such a short half‐life explains the high doses of allatostatins required to obtain the expected biological effects when tested in ...

Cardiac rhythm was measured in Blattella germanica females during the reproductive cycle. The rate increased from day 0 to 1, remained constant during the vitellogenic period and fell by about 20% during the period of oothecal transport. The effects of allatostatins, allatostatin analogues and corazonin were tested on semi-isolated heart preparations. Allatostatins showed a rapid, reversible and dose-dependent cardioinhibitory activity. Blattella allatostatin 1 (BLAST-1: LYDFGL-NH 2 ), was the most active, eliciting 76% inhibition at 10 Ϫ7 M and even 19% inhibition at 10 Ϫ9 M. BLAST-2 (DRLYSFGL-NH 2 ), BLAST-3 (AGSDGRLYSFGL-NH 2 ) and BLAST-4 (APSSAQRLYGFGL-NH 2 ) were less active. An analogue of BLAST-2 with C-terminus in acid form and a pseudopeptide analogue of BLAST-2 with a methyleneamino ⌿[CH 2 NH] peptide bond surrogate between residues L 3 and Y 4 were inactive. Corazonin elicited rapid, reversible and dose-dependent cardioacceleratory activity. When tested together with BLAST-1, corazonin overrode the cardioinhibitory effect of allatostatin. Our previous results had shown that high levels of allatostatin were maintained during the period of oothecal transport. This and the fact that physiological concentrations of allatostatins produce physiological levels of inhibition, suggest that allatostatins are involved in the modulation of cardiac rhythm in this cockroach.

The diamide insecticide tetraniliprole is a valuable tool for managing major insect pests like the invasive tomato pinworm, Tuta absoluta (Meyrick). However, the mechanisms underlying tetraniliprole resistance, as well as its associated fitness costs, remain unclear. In this study, we assessed the fitness of tetraniliprole-resistant (TetraRS) and susceptible (SS) strains of T. absoluta and conducted Illumina RNA-seq to compare their transcriptomes. We also used nanocarrier-mediated RNA interference (RNAi) to knockdown P450 genes and evaluate their role in tetraniliprole resistance. After eight generations of selection, T. absoluta developed a 20.80-fold resistance to tetraniliprole, accompanied by fitness costs. RNA-seq analysis revealed 3332 differentially expressed genes (DEGs), with 1707 upregulated and 1625 downregulated in the TetraRS compared to the SS strain. Gene Ontology (GO) annotations showed significant enrichment in categories related to metabolic processes, cellular processes, catalytic activity, cellular anatomical entity, and binding. These genes were also identified in key KEGG pathways such as cytochrome P450, drug metabolism, carbon metabolism, oxidative phosphorylation, fatty acid metabolism, and protein processing. RT-qPCR analysis confirmed that P450 genes (CYP405D1, CYP6AB269, and CYP4AU1) were upregulated in TetraRS insects, in line with the RNA-seq results. Cytochrome P450 activity was significantly higher in the TetraRS strain than in the SS strain. Notably, nanoencapsulated dsRNA targeting these overexpressed P450 genes increased the susceptibility of T. absoluta to tetraniliprole. Further, cytochrome P450 activity was significantly reduced following silencing of P450 genes. These findings suggest that multiple genes and pathways, particularly P450 genes, contribute to tetraniliprole resistance in T. absoluta. This study provides valuable insights into the molecular mechanisms underlying insecticide resistance in this key pest species.

Nitrogen fixation in a wood-feeding termite, Neotermes koshunensis 4. From wood-to soil-feeding termites: A study in a tropical forest in Cameroon, central Africa Diversification and evolution of humivore in Termes-Capritermes branch of the subfamily Termitinae in Darwin, northern Australia ••••39 6. Two short notes of (i 13 C-(i 15 N natural abundance method 6.1 Fungus growing termites in Cameroon, Thailand and Cote d'Ivore 6.2 Wood-feeders in Sydney and grass-harvesters in Townsville 7. General discussion 7.1 Nitrogen fixation in various termites -its pattern and implication by applying (i 15 N natural abundance method 55 7.2 Stable isotope ratios in detritivorous animals 7.3 Termite-symbionts system by stable isotope techniques 8. Conclusion Acknowledgments Synopsis (In Japanese : 1~~)

Climate change is affecting biodiversity, but proximate drivers remain poorly understood. Here, we examine how experimental heatwaves impact on reproduction in an insect system. Male sensitivity to heat is recognised in endotherms, but ectotherms have received limited attention, despite comprising most of biodiversity and being more influenced by temperature variation. Using a flour beetle model system, we find that heatwave conditions (5 to 7 °C above optimum for 5 days) damaged male, but not female, reproduction. Heatwaves reduce male fertility and sperm competitiveness, and successive heatwaves almost sterilise males. Heatwaves reduce sperm production, viability, and migration through the female. Inseminated sperm in female storage are also damaged by heatwaves. Finally, we discover transgenerational impacts, with reduced reproductive potential and lifespan of offspring when fathered by males, or sperm, that had experienced heatwaves. This male reproductive damage under heatwave ...

Nurturing, especially feeding ant colonies is a vital topic among ant researchers and ant keepers. Numerous factors contribute to growing healthy ant colonies rapidly, including, for instance, humidity levels and appropriate farming methods. Proper diet is also an essential consideration. Many articles contemplate the potential of artificial diets. Variable carbohydrate-based diets have been tested but the role of protein has largely been overlooked. We aimed to analyse the effects of these diets to be able to standardize and compare results of different laboratories. We assessed the most readily available, natural, inexpensive, known, and efficient diets to ensure optimal nutrition for ant colonies by analysing the development of 100 Lasius niger colonies. They were given four different carbohydrate-based diets in the first year and four different protein-based diets in the second year. Based on our findings, honey water was the best carbohydrate source. The best protein diet, however, consisted of crickets, which positively influenced not only the development but also the survival success of the colonies during the wintering period. Thus, we suggest rearing laboratory colonies of L. niger on honey water and crickets. It would be worth repeating this study with different ant species and doing biochemical analysis on the diets.

Powderpost beetles such as Lyctus africanus are a common pest group for dried cured wood, causing significant harm to wood and wood products. We examined the life span and effects of aging and mating status on pheromone production in the powderpost beetle L. africanus (Coleoptera: Lyctinae). Experiments compared starved and unstarved male groups, and chemical analysis was used to determine factors affecting pheromone production. Regarding lifespan, male beetles provided food survived up to 14 weeks, while starved beetles died before the fifth week. Thus, an adult L. africanus male may require food throughout its lifespan, and food availability may affect pheromone production. There was no significant difference in the quantity of two major pheromone compounds, compound 2 (3-pentyl dodecanoate) and 3 (3-pentyl tetradecanoate) between mated and un-mated males. On the other hand, a minor compound, compound 1 (2-propyl dodecanoate) showed increased quantity after mating. The two major c...

In polyandrous insects, postcopulatory sexual selection is a pervasive evolutionary force favouring male and female traits that allow control of offspring paternity. Males may influence paternity through adaptations for sperm competition, and females through adaptations facilitating cryptic female choice. Yet, the mechanisms are often complex, involving behaviour, physiology or morphology, and they are difficult to identify. In red flour beetles (Tribolium castaneum), paternity varies widely, and evidence suggests that both male and female traits influence the outcome of sperm competition. To test the role of spermathecal morphology and of sperm storage processes on the outcome of sperm competition, we mated each of 26 virgin females with two males, one of which carrying a phenotypic marker to assign offspring paternity. We manipulated the interval between mating with the first and the second male, to create different conditions of sperm storage (overlapping and nonoverlapping) in the female reproductive tract. To investigate the role of sperm storage more closely, we examined the relationship between paternity and spermathecal morphology in a subset of 14 experimental females. In addition, we also characterized variation in spermathecal morphology in three different strains, wildtype, Chicago black and Reindeer. No significant influence of the intermating interval was found on the paternity of the focal male, although the direction of the difference was in the expected direction of higher last male paternity for longer intervals. Moreover, paternity was not significantly associated with spermathecal morphology, although spermathecal volume, complexity, and tubule width varied significantly and substantially among individuals in all investigated strains.

The multicoloured Asian ladybirds, Harmonia axyridis (Pallas), form large aggregations inside dwellings to survive cold winters, causing annoyance to householders due to their abundance, allergenic properties and problems consequent on reflex bleeding. Flight to overwintering sites and macrosite choice of this species is well documented. H. axyridis shows a hypsotactic behaviour and a clear preference for contrasting visual elements. However, how the microsite is selected remains undocumented, although a better understanding of the factors implicated could lead to the development of new control methods for this pest. In this work, we hypothesised that non-volatile compounds are involved in the microsite choice and aggregation of this beetle. Long chain hydrocarbons were identified at aggregation sites, comprising saturated and unsaturated homologues. An aggregation bioassay was then conducted on overwintering individuals, highlighting the retention capacity of the identified compounds on the tested ladybirds. Additional investigations showed that H. axyridis males and females, originating from overwintering sites, deposit a similar blend of molecules while walking. A Y-shaped tube assay revealed that this blend is used by male and female congeners as cue allowing individuals to orientate towards the treated side of the olfactometer. These results suggest the use of two different blends of long chain hydrocarbons by H. axyridis during its aggregative period, the first one to lead conspecifics towards aggregation sites (microsites) and the second to ensure the cohesion of the aggregation. These findings support the potential use of these blends, in association with volatiles, in the design of traps in order to control infestations of this species in dwellings.