Strong resistance to ß-cyfluthrin in a strain of the beetle Alphitobius diaperinus: a de novo transcriptome analysis.

The lesser mealworm, Alphitobius diaperinus, is an invasive tenebrionid beetle and a vector of pathogens. Due to the emergence of insecticide resistance and consequent outbreaks that generate significant phytosanitary and energy costs for poultry farmers, it has become a major insect pest worldwide. To better understand the molecular mechanisms behind this resistance, we studied a strain of A. diaperinus from a poultry house in Brittany that was found to be highly resistant to the ß-cyfluthrin. The strain survived ß-cyfluthrin exposures corresponding to more than 100 times the recommended dose. We used a comparative de novo RNA-Seq approach to explore genes expression in resistant versus sensitive strains. Our de novo transcriptomic analyses showed that responses to ß-cyfluthrin likely involved a whole set of resistance mechanisms. Genes related to detoxification, metabolic resistance, cuticular hydrocarbon biosynthesis and proteolysis were found to be constitutively overexpressed in the resistant compared to the sensitive strain. Follow-up enzymatic assays confirmed that the resistant strain exhibited high basal activities for detoxification enzymes such as cytochrome P450 monooxygenase and glutathione-S-transferase. The in-depth analysis of differentially expressed genes suggests the involvement of complex regulation of signaling pathways. Detailed knowledge of these resistance mechanisms is essential for the establishment of effective pest control.

The Plasticene era: Current uncertainties in estimates of the hazards posed by tiny plastic particles on soils and terrestrial invertebrates.

Plastics are ubiquitous in our daily life. Large quantities of plastics leak in the environment where they weather and fragment into micro- and nanoparticles. This potentially releases additives, but rarely leads to a complete mineralization, thus constitutes an environmental hazard. Plastic pollution in agricultural soils currently represents a major challenge: quantitative data of nanoplastics in soils as well as their effects on biodiversity and ecosystem functions need more attention. Plastic accumulation interferes with soil functions, including water dynamics, aeration, microbial activities, and nutrient cycling processes, thus impairing agricultural crop yield. Plastic debris directly affects living organisms but also acts as contaminant vectors in the soils, increasing the effects and the threats on biodiversity. Finally, the effects of plastics on terrestrial invertebrates, representing major taxa in abundance and diversity in the soil compartment, need urgently more investigation from the infra-individual to the ecosystem scales.

Exploring cross-protective effects between cold and immune stress in Drosophila melanogaster.

It is well established that environmental and biotic stressors like temperature and pathogens/parasites are essential for the life of small ectotherms. There are complex interactions between cold stress and pathogen infection in insects. Possible cross-protective mechanisms occur between both stressors, suggesting broad connectivity in insect stress responses. In this study, the functional significance of these interactions was tested, as well as the potential role of newly uncovered candidate genes, turandot. This was done using an array of factorial experiments exposing Drosophila melanogaster flies to a combination of different cold stress regimes (acute or chronic) and infections with the parasitic fungus Beauveria bassiana. Following these crossed treatments, phenotypic and molecular responses were assessed by measuring 1) induced cold tolerance, 2) immune resistance to parasitic fungus, and 3) activation of turandot genes. We found various responses in the phenotypic outcomes according to the various treatment combinations with higher susceptibility to infection following cold stress, but also significantly higher acute cold survival in flies that were infected. Regarding molecular responses, we found overexpression of turandot genes in response to most treatments, suggesting reactivity to both cold and infection. Moreover, maximum peak expressions were distinctly observed in the combined treatments (infection plus cold), indicating a marked synergistic effect of the stressors on turandot gene expression patterns. These results reflect the great complexity of cross-tolerance reactions between infection and abiotic stress, but could also shed light on the mechanisms underlying the activation of these responses.

Title: Exploration des effets de protection croisée entre le froid et le stress immunitaire chez Drosophila melanogaster. Abstract: Il est établi que les stress environnementaux et biotiques, tels que la température et les agents pathogènes ou parasites, sont essentiels à la survie des petits ectothermes. Il existe des interactions complexes entre le stress froid et l'infection par des pathogènes chez les insectes. Des mécanismes de protection croisée sont possibles entre les deux facteurs de stress, suggérant une grande connectivité entre ces réponses. Dans cette étude, ces interactions ont été testées, ainsi que l'implication potentielle de nouveaux gènes candidats, les gènes turandot. Cela a été fait via un ensemble d'expériences factorielles exposant des mouches Drosophila melanogaster à différents types de stress froid (aigu ou chronique) couplés à des infections par le champignon entomoparasite Beauveria bassiana. Suite à ces traitements croisés, les réponses phénotypiques et moléculaires ont été analysées en mesurant 1) la tolérance induite au froid, 2) la résistance immunitaire à l'infection pathogène et 3) l'activation des gènes turandot. Les résultats phénotypiques ont montré des réponses variées selon les différentes combinaisons de traitements, avec une plus grande susceptibilité à l'infection suite à un stress froid, mais aussi une survie au stress froid aigu significativement plus élevée chez les mouches infectées. Pour les réponses moléculaires, nous avons trouvé une surexpression des gènes turandot en réponse à la plupart des traitements, suggérant une réactivité à la fois au froid et à l'infection. De plus, les expressions maximales ont été distinctivement observées dans les traitements combinés (infection plus froid), indiquant un effet synergique marqué des facteurs de stress sur l'expression des gènes turandot. Ces résultats reflètent la complexité des réactions de tolérance croisée entre infection et stress abiotique, et pourraient également éclairer les mécanismes sous-jacents de l'activation de ces réponses.

Preferred temperature in the warmth of cities: Body size, sex and development stage matter more than urban climate in a ground-dwelling spider.

Most ectotherms rely on behavioural thermoregulation to maintain body temperatures close to their physiological optimum. Hence, ectotherms can drastically limit their exposure to thermal extremes by selecting a narrower range of temperatures, which includes their preferred temperature (Tpref). Despite evidence that behavioural thermoregulation can be adjusted by phenotypic plasticity or constrained by natural selection, intraspecific Tpref variations across environmental gradients remain overlooked as compared to other thermal traits like thermal tolerance. Here, we analyzed Tpref variation of spider populations found along a gradient of urban heat island (UHI) which displays large thermal variations over small distances. We measured two components of the thermal preference, namely the mean Tpref and the Tpref range (i.e., standard deviation) in 557 field-collected individuals of a common ground-dwelling spider (Pardosa saltans, Lycosidae) using a laboratory thermal gradient. We determined if Tpref values differed among ten populations from contrasting thermal zones. We showed that endogenous factors such as body size or sex primarily determine both mean Tpref and Tpref range. The Tpref range was also linked to the UHI intensity to a lesser extent, yet only in juveniles. The absence of relationship between Tpref metrics and UHI in adult spiders suggests a Bogert effect according to which the ability of individuals to detect and exploit optimal microclimates weakens the selection pressure of temperatures (here driven by UHI) on their thermal physiology. Alternatively, this lack of relationship could also indicate that temperature patterns occurring at the scale of the spiders' micro-habitat differ from measured ones. This study shows the importance of considering both inter-individual and inter-population variations of the Tpref range when conducting Tpref experiments, and supports Tpref range as being a relevant measure to inform on the strength of behavioural thermoregulation in a given population.

Mitochondrial metabolism and oxidative stress in the tropical cockroach under fluctuating thermal regimes.

The cockroach Gromphadorhina coquereliana can survive at low temperatures under extensive periods of cold stress. To assess energy management and insect adaptation in response to cold, we measured mitochondrial activity and oxidative stress in muscle and fat body tissues from G. coquereliana under a fluctuating thermal regime (FTR; stressed at 4°C for 3 h on 3 consecutive days, with or without 24 h recovery). Compared with our earlier work showing that a single exposure to cold significantly affects mitochondrial parameters, here, repeated exposure to cold triggered an acclimatory response, resulting in unchanged mitochondrial bioenergetics. Immediately after cold exposure, we observed an increase in the overall pool of ATP and a decrease in typical antioxidant enzyme activity. We also observed decreased activity of uncoupling protein 4 in muscle mitochondria. After 24 h of recovery, we observed an increase in expression of antioxidant enzymes in muscles and the fat body and a significant increase in the expression of UCP4 and HSP70 in the latter. This indicates that processes related to energy conversion and disturbance under cold stress may trigger different protective mechanisms in these tissues, and that these mechanisms must be activated to restore insect homeostasis. The mitochondrial parameters and enzymatic assays suggest that mitochondria are not affected during FTR but oxidative stress markers are decreased, and a 24 h recovery period allows for the restoration of redox and energy homeostasis, especially in the fat body. This confirms the crucial role of the fat body in intermediary metabolism and energy management in insects and in the response to repeated thermal stress.

Interspecific differences in thermal tolerance landscape explain aphid community abundance under climate change.

A single critical thermal limit is often used to explain and infer the impact of climate change on geographic range and population abundance. However, it has limited application in describing the temporal dynamic and cumulative impacts of extreme temperatures. Here, we used a thermal tolerance landscape approach to address the impacts of extreme thermal events on the survival of co-existing aphid species (Metopolophium dirhodum, Sitobion avenae and Rhopalosiphum padi). Specifically, we built the thermal death time (TDT) models based on detailed survival datasets of three aphid species with three ages across a broad range of stressful high (34-40 °C) and low (-3∼-11 °C) temperatures to compare the interspecific and developmental stage variations in thermal tolerance. Using these TDT parameters, we performed a thermal risk assessment by calculating the potential daily thermal injury accumulation associated with the regional temperature variations in three wheat-growing sites along a latitude gradient. Results showed that M. dirhodum was the most vulnerable to heat but more tolerant to low temperatures than R. padi and S. avenae. R. padi survived better at high temperatures than Sitobion avenae and M. dirhodum but was sensitive to cold. R. padi was estimated to accumulate higher cold injury than the other two species during winter, while M. dirhodum accrued more heat injury during summer. The warmer site had higher risks of heat injury and the cooler site had higher risks of cold injury along a latitude gradient. These results support recent field observations that the proportion of R. padi increases with the increased frequency of heat waves. We also found that young nymphs generally had a lower thermal tolerance than old nymphs or adults. Our results provide a useful dataset and method for modelling and predicting the consequence of climate change on the population dynamics and community structure of small insects.

Metabolomics as a tool to elucidate biochemical cold adaptation in insects.

Metabolomics is an incredibly valuable tool in helping understand insect responses to cold. It not only characterizes how low temperature disrupts metabolic homeostasis, but also how it triggers fundamental adaptive responses, for example, homeoviscous adaptation and cryoprotectant accumulation. This review outlines the advantages and disadvantages of different metabolomic technologies (nuclear magnetic resonance- versus mass spectrometry-based) and screening approaches (targeted versus untargeted). We emphasize the importance of time-series and tissue-specific data, as well as the challenges of disentangling insect versus microbiome responses. In addition, we set out the need to move beyond simple correlations between metabolite abundance and tolerance phenotypes by undertaking functional assessments, for example, using dietary supplementation or injections. We highlight studies at the vanguard of employing these approaches, and where key knowledge gaps remain.

The interspecific variations in molecular responses to various doses of heat and cold stress: The case of cereal aphids.

Insects are currently subjected to unprecedented thermal stress due to recent increases in the frequency and amplitude of temperature extremes. Understanding molecular responses to thermal stress is critically important to appreciate how species react to thermal stress. Three co-occurring cosmopolitan species are found within the guild of cereal aphids: Sitobion avenae, Ropalosiphum padi and Metopolophium dirhodum. Earlier reports have shown that increasing frequency of temperature extremes causes a shift in dominant species within guilds of cereal aphids by differently altering the population's growth. We hypothesize that a differential molecular response to stress among species may partially explain these changes. Heat shock proteins (HSPs) are molecular chaperones well known to play an important role in protecting against the adverse effects of thermal stress. However, few studies on molecular chaperones have been conducted in cereal aphids. In this study, we compared the heat and cold tolerance between three aphid species by measuring the median lethal time (Lt50) and examined the expression profiles of seven hsp genes after exposures to comparable thermal injury levels and also after same exposure durations. Results showed that R. padi survived comparatively better at high temperatures than the two other species but was more cold-sensitive. Hsp genes were induced more strongly by heat than cold stress. Hsp70A was the most strongly up-regulated gene in response to both heat and cold stress. R. padi had more heat inducible genes and significantly higher mRNA levels of hsp70A, hsp10, hsp60 and hsp90 than the other two species. Hsps ceased to be expressed at 37 °C in M. dirhodum and S. avenae while expression was maintained in R. padi. In contrast, M. dirhodum was more cold tolerant and had more cold inducible genes than the others. These results confirm species-specific differences in molecular stress responses and suggest that differences in induced expression of hsps may be related to species' thermal tolerance, thus causing the changes in the relative abundance.

The transcriptomic signature of responses to larval crowding in Drosophila melanogaster.

Intraspecific competition at the larval stage is an important ecological factor affecting life-history, adaptation and evolutionary trajectory in holometabolous insects. However, the molecular pathways underpinning these ecological processes are poorly characterized. We reared Drosophila melanogaster at three egg densities (5, 60, and 300 eggs/mL) and sequenced the transcriptomes of pooled third-instar larvae. We also examined emergence time, egg-to-adult viability, adult mass, and adult sex-ratio at each density. Medium crowding had minor detrimental effects on adult phenotypes compared to low density and yielded 24 differentially expressed genes (DEGs), including several chitinase enzymes. In contrast, high crowding had substantial detrimental effects on adult phenotypes and yielded 2107 DEGs. Among these, upregulated gene sets were enriched in sugar, steroid and amino acid metabolism as well as DNA replication pathways, whereas downregulated gene sets were enriched in ABC transporters, taurine, Toll/Imd signaling, and P450 xenobiotics metabolism pathways. Overall, our findings show that larval crowding has a large consistent effect on several molecular pathways (i.e., core responses) with few pathways displaying density-specific regulation (i.e., idiosyncratic responses). This provides important insights into how holometabolous insects respond to intraspecific competition during development.

The rising threat of climate change for arthropods from Earth's cold regions: Taxonomic rather than native status drives species sensitivity.

Polar and alpine regions are changing rapidly with global climate change. Yet, the impacts on biodiversity, especially on the invertebrate ectotherms which are dominant in these areas, remain poorly understood. Short-term extreme temperature events, which are growing in frequency, are expected to have profound impacts on high-latitude ectotherms, with native species being less resilient than their alien counterparts. Here, we examined in the laboratory the effects of short periodic exposures to thermal extremes on survival responses of seven native and two non-native invertebrates from the sub-Antarctic Islands. We found that survival of dipterans was significantly reduced under warming exposures, on average having median lethal times (LT50 ) of about 30 days in control conditions, which declined to about 20 days when exposed to daily short-term maxima of 24°C. Conversely, coleopterans were either not, or were less, affected by the climatic scenarios applied, with predicted LT50 as high as 65 days under the warmest condition (daily exposures at 28°C for 2 h). The native spider Myro kerguelensis was characterized by an intermediate sensitivity when subjected to short-term daily heat maxima. Our results unexpectedly revealed a taxonomic influence, with physiological sensitivity to heat differing between higher level taxa, but not between native and non-native species representing the same higher taxon. The survival of a non-native carabid beetle under the experimentally imposed conditions was very high, but similar to that of native beetles, while native and non-native flies also exhibited very similar sensitivity to warming. As dipterans are a major element of diversity of sub-Antarctic, Arctic and other cold ecosystems, such observations suggest that the increased occurrence of extreme, short-term, thermal events could lead to large-scale restructuring of key terrestrial ecosystem components both in ecosystems protected from and those exposed to the additional impacts of biological invasions.

Dietary salt supplementation adversely affects thermal acclimation responses of flight ability in Drosophila melanogaster.

Cold acclimation may enhance low temperature flight ability, and salt loading can alter an insects' cold tolerance by affecting their ability to maintain ion balance in the cold. Presently however, it remains unclear if dietary salt impacts thermal acclimation of flight ability in insects. Here, we examined the effect of a combination of dietary salt loading (either NaCl or KCl) and low temperature exposure on the flight ability of Drosophila melanogaster at low (15 °C) and benign (optimal, 22 °C) temperatures. Additionally, we determined whether dietary salt supplementation translates into increased K+ and Na+ levels in the bodies of D. melanogaster. Lastly, we determined whether salt supplementation impacts body mass and wing morphology, to ascertain whether any changes in flight ability were potentially driven by flight-related morphometric variation. In control flies, we find that cold acclimation enhances low temperature flight ability over non-acclimated flies confirming the beneficial acclimation hypothesis. By contrast, flies supplemented with KCl that were cold acclimated and tested at a cold temperature had the lowest flight ability, suggesting that excess dietary KCl during development negates the beneficial cold acclimation process that would have otherwise taken place. Overall, the NaCl-supplemented flies and the control group had the greatest flight ability, whilst those fed a KCl-supplemented diet had the lowest. Dietary salt supplementation translated into increased Na+ and K+ concentration in the body tissues of flies, confirming that dietary shifts are reflected in changes in body composition and are not simply regulated out of the body by homeostasis over the course of development. Flies fed with a KCl-supplemented diet tended to be larger with larger wings, whilst those reared on the control or NaCl-supplemented diet were smaller with smaller wings. Additionally, the flies with greater flight ability tended to be smaller and have lower wing loading. In conclusion, dietary salts affected wing morphology as well as ion balance, and dietary KCl seemed to have a detrimental effect on cold acclimation responses of flight ability in D. melanogaster.

Rapid Adjustments in Thermal Tolerance and the Metabolome to Daily Environmental Changes - A Field Study on the Arctic Seed Bug Nysius groenlandicus.

Laboratory investigations on terrestrial model-species, typically of temperate origin, have demonstrated that terrestrial ectotherms can cope with daily temperature variations through rapid hardening responses. However, few studies have investigated this ability and its physiological basis in the field. Especially in polar regions, where the temporal and spatial temperature variations can be extreme, are hardening responses expected to be important. Here, we examined diurnal adjustments in heat and cold tolerance in the Greenlandic seed bug Nysius groenlandicus by collecting individuals for thermal assessment at different time points within and across days. We found a significant correlation between observed heat or cold tolerance and the ambient microhabitat temperatures at the time of capture, indicating that N. groenlandicus continuously and within short time-windows respond physiologically to thermal changes and/or other environmental variables in their microhabitats. Secondly, we assessed underlying metabolomic fingerprints using GC-MS metabolomics in a subset of individuals collected during days with either low or high temperature variation. Concentrations of metabolites, including sugars, polyols, and free amino acids varied significantly with time of collection. For instance, we detected elevated sugar levels in animals caught at the lowest daily field temperatures. Polyol concentrations were lower in individuals collected in the morning and evening and higher at midday and afternoon, possibly reflecting changes in temperature. Additionally, changes in concentrations of metabolites associated with energetic metabolism were observed across collection times. Our findings suggest that in these extreme polar environments hardening responses are marked and likely play a crucial role for coping with microhabitat temperature variation on a daily scale, and that metabolite levels are actively altered on a daily basis.

Acclimation, duration and intensity of cold exposure determine the rate of cold stress accumulation and mortality in Drosophila suzukii.

The spotted wing drosophila (SWD), Drosophila suzukii, is a major invasive fruit pest. There is strong consensus that low temperature is among the main drivers of SWD population distribution, and the invasion success of SWD is also linked to its thermal plasticity. Most studies on ectotherm cold tolerance focus on exposure to a single stressful temperature but here we investigated how cold stress intensity affected survival duration across a broad range of low temperatures (-7 to +3 °C). The analysis of Lt50 at different stressful temperatures (Thermal Death Time curve - TDT) is based on the suggestion that cold injury accumulation rate increases exponentially with the intensity of thermal stress. In accordance with the hypothesis, Lt50 of SWD decreased exponentially with temperature. Further, comparison of TDT curves from flies acclimated to 15, 19 and 23 °C, respectively, showed an almost full compensation with acclimation such that the temperature required to induce mortality over a fixed time decreased almost 1 °C per °C lowering of acclimation temperature. Importantly, this change in cold tolerance with acclimation was uniform across the range of moderate to intense cold stress exposures examined. To understand if cold stress at moderate and intense exposures affects the same physiological systems we examined how physiological markers/symptoms of chill injury developed at different intensities of the cold stress. Specifically, hsp23 expression and extracellular [K+] were measured in flies exposed to different intensities of cold stress (-6, -2 and +2 °C) and at various time points corresponding to the same progression of injury (equivalent to 1/3, 2/3 or 3/3 of Lt50). The different cold stress intensities all triggered hsp23 expression following 2 h of recovery, but patterns of expression differed. At the most intense cold stress (-6 and -2 °C) a gradual increase with time was found. In contrast, at +2 °C an initial increase was followed by a dissipating expression. A gradual perturbation of ion balance (hyperkalemia) was also found at all three cold stress intensities examined, with only slight dissimilarities between treatment temperatures. Despite some differences between the three cold intensities examined, the results generally support the hypothesis that intense and moderate cold stress induces the same physiological perturbation. This suggests that cold stress experienced during natural fluctuating conditions is additive and the results also illustrate that the rate of injury accumulation increases dramatically (exponentially) with decreasing temperature (increasing stress).

The discovery, distribution, and diversity of DNA viruses associated with Drosophila melanogaster in Europe.

Drosophila melanogaster is an important model for antiviral immunity in arthropods, but very few DNA viruses have been described from the family Drosophilidae. This deficiency limits our opportunity to use natural host-pathogen combinations in experimental studies, and may bias our understanding of the Drosophila virome. Here, we report fourteen DNA viruses detected in a metagenomic analysis of 6668 pool-sequenced Drosophila, sampled from forty-seven European locations between 2014 and 2016. These include three new nudiviruses, a new and divergent entomopoxvirus, a virus related to Leptopilina boulardi filamentous virus, and a virus related to Musca domestica salivary gland hypertrophy virus. We also find an endogenous genomic copy of galbut virus, a double-stranded RNA partitivirus, segregating at very low frequency. Remarkably, we find that Drosophila Vesanto virus, a small DNA virus previously described as a bidnavirus, may be composed of up to twelve segments and thus represent a new lineage of segmented DNA viruses. Two of the DNA viruses, Drosophila Kallithea nudivirus and Drosophila Vesanto virus are relatively common, found in 2 per cent or more of wild flies. The others are rare, with many likely to be represented by a single infected fly. We find that virus prevalence in Europe reflects the prevalence seen in publicly available datasets, with Drosophila Kallithea nudivirus and Drosophila Vesanto virus the only ones commonly detectable in public data from wild-caught flies and large population cages, and the other viruses being rare or absent. These analyses suggest that DNA viruses are at lower prevalence than RNA viruses in D.melanogaster, and may be less likely to persist in laboratory cultures. Our findings go some way to redressing an earlier bias toward RNA virus studies in Drosophila, and lay the foundation needed to harness the power of Drosophila as a model system for the study of DNA viruses.

Divergent metabolomic profiles of cold-exposed mature and immature females of tropical versus temperate Drosophila species.

Temperate species, contrary to their tropical counterparts, are exposed not only to thermally variable environments with low temperatures but also to long winters. Different selective pressures may have driven divergent physiological adaptations in closely related species with different biogeographic origins. To survive unfavourable winter conditions, Drosophila species in temperate areas generally undergo a period of reproductive dormancy, associated with a cold-induced cessation of oogenesis and metabolic reorganization. This work aims to compare cold tolerance and metabolic signatures of cold-exposed females exhibiting different reproductive maturity status (mature and immature females) of four Drosophila species from tropical vs. temperate origins. We expected that the capacity for delayed reproduction of immature females could result in the redirection of the energy-related metabolites to be utilized for surviving the cold season. To do so, we studied an array of 45 metabolites using quantitative target GC-MS profiling. Reproductively immature females of temperate species showed the lower CTmin and the faster chill coma recovery time (i.e. the most cold-tolerant group). Principal component analysis captured differences across species, but also between reproductive maturity states. Notably, temperate species exhibited significantly higher levels of glucose, alanine, and gluconolactone than tropical ones. As proline and glycerol showed higher abundances in immature females of temperate species compared to the levels exhibited by the rest of the groups, we reasoned that glucose and alanine could serve as intermediates in the synthesis of these compatible solutes. All in all, our findings suggest that cold-exposed females of temperate species accumulate energy-related and protective metabolites (e.g. glycerol and proline) while delaying reproduction, and that these metabolites are relevant to cold tolerance even at modest concentrations.

Thermal plasticity and sensitivity to insecticides in populations of an invasive beetle: Cyfluthrin increases vulnerability to extreme temperature.

Climate change increases average temperatures and the occurrence of extreme weather events, in turn accentuating the risk of organism exposure to temperature stress. When thermal conditions become stressful, the sensitivity of insects toward insecticides can be exacerbated. Likewise, exposure of insects to insecticides can subsequently influence their ability to handle stressful temperatures. Here, we investigated the effects of constant temperature and daily heat spikes, in presence/absence of insecticide treatment (cyfluthrin), on the condition (impairment of mobility) and thermal tolerance to cold (-6 °C) and heat (42.5 °C) of the terrestrial beetle Alphitobius diaperinus. The responses of insects from four populations (three farm-collected populations, one laboratory population) to different durations of extreme temperature exposure were compared. The results showed that the laboratory population was generally more sensitive to extreme cold and heat temperatures, with less than 50% of adults recovering after an exposure at -6 or +42.5 °C for 3h. Significant differences in the level of thermal tolerance were also found among insects from poultry farms. Cyfluthrin exposure incurred detrimental effects to insects' condition in all but one population. For two out of the four populations, mobility impairment was increased when adults were exposed to daily heat spikes (6 h per day at 38 °C) and cyfluthrin simultaneously, compared to cyfluthrin exposure at constant temperatures; yet, no significant interaction between the two stressors was found. Finally, using one farm collected population, effects of pre-exposure to cyfluthrin on extreme temperature tolerance provided another example of the toxicant-induced climate sensitivity in insects.

Biogeographic position and body size jointly set lower thermal limits of wandering spiders.

Most species encounter large variations in abiotic conditions along their distribution range. The physiological responses of most terrestrial ectotherms (such as insects and spiders) to clinal gradients of climate, and in particular gradients of temperature, can be the product of both phenotypic plasticity and local adaptation. This study aimed to determine how the biogeographic position of populations and the body size of individuals set the limits of cold (freezing) resistance of Dolomedes fimbriatus. We compared D. fimbriatus to its sister species Dolomedes plantarius under harsher climatic conditions in their distribution range. Using an ad hoc design, we sampled individuals from four populations of Dolomedes fimbriatus originating from contrasting climatic areas (temperate and continental climate) and one population of the sister species D. plantarius from continental climate, and compared their supercooling ability as an indicator of cold resistance. Results for D. fimbriatus indicated that spiders from northern (continental) populations had higher cold resistance than spiders from southern (temperate) populations. Larger spiders had a lower supercooling ability in northern populations. The red-listed and rarest D. plantarius was slightly less cold tolerant than the more common D. fimbriatus, and this might be of importance in a context of climate change that could imply colder overwintering habitats in the north due to reduced snow cover protection. The lowest cold resistance might put D. plantarius at risk of extinction in the future, and this should be considered in conservation plan.

Differences in the Susceptibility to Commercial Insecticides among Populations of the Lesser Mealworm Alphitobius diaperinus Collected from Poultry Houses in France.

The control of insect pests often relies on the recurrent application of insecticides. This is the case for the lesser mealworm, Alphitobius diaperinus, an invasive beetle infesting poultry farms. There is evidence that A. diaperinus can develop resistance to several insecticides. Evaluation of such resistance has never been conducted in France, despite the beetle's presence since the 1970s. We assess insecticide susceptibility in 10 populations from French poultry farms and compare patterns with two susceptible populations. Adults are subjected to short-term exposures (4 h) to four commercial insecticides and their recovery is assessed. Temporal survival also is scored during constant exposures for seven days. Clear-cut differences among the farm populations are found. Except for three populations that have patterns similar to those of the two susceptible populations, all the other farm populations have a much greater capacity to recover and survive insecticide exposures, especially to pyrethroid-based formulations. Three populations in particular even exhibit clear signs of resistance to pyrethroids, with median lethal times more than 10-fold superior to values of the susceptible population. No insect in any population recovers from the pirimiphos-methyl exposure, and all beetles are apparently dead after 15 h. Our results demonstrate the existence of resistant populations to pyrethroids in Brittany France.

Cell free Microcystis aeruginosa spent medium affects Daphnia magna survival and stress response.

Primary consumers in freshwater ecosystems, such as the zooplankton organism Daphnia magna, are highly affected by cyanobacteria, both as they may use it as a food source but also by cyanobacterial metabolites present in the water. Here, we investigate the impacts of cyanobacterial metabolites focussing on the environmental realistic scenario of the naturally released mixture without crushing cyanobacterial cells or their uptake as food. Therefore, D. magna were exposed to two concentrations of cell free cyanobacterial spent medium from Microcystis aeruginosa PCC 7806 to represent higher and lower ecologically-relevant concentrations of cyanobacterial metabolites. Including microcystin-LR, 11 metabolites have been detected of which 5 were quantified. Hypothesising concentration and time dependent negative impact, survival, gene expression marking digestion and metabolism, oxidative stress response, cell cycle and molting as well as activities of detoxification and antioxidant enzymes were followed for 7 days. D. magna suffered from oxidative stress as both catalase and glutathione S-transferase enzyme activities significantly decreased, suggesting enzyme exhaustibility after 3 and 7 days. Moreover, gene-expressions of the 4 stress markers (glutathione S-transferase, glutathione peroxidase, catalase and thioredoxin) were merely downregulated after 7 days of exposure. Energy allocation (expression of glyceraldehyde-3-phosphate dehydrogenase) was increased after 3 days but decreased as well after 7 days exposure. Cell cycle was impacted time dependently but differently by the two concentrations, along with an increasing downregulation of myosin heavy chain responsible for cell arrangement and muscular movements. Deregulation of nuclear hormone receptor genes indicate that D. magna hormonal steering including molting seemed impaired despite no detection of microviridin J in the extracts. As a consequence of all those responses and presumably of more than investigated molecular and physiological changes, D. magna survival was impaired over time, in a concentration dependent manner. Our results confirm that besides microcystin-LR, other secondary metabolites contribute to negative impact on D. magna survival and stress response.

Role of the Insect Neuroendocrine System in the Response to Cold Stress.

Insects are the largest group of animals. They are capable of surviving in virtually all environments from arid deserts to the freezing permafrost of polar regions. This success is due to their great capacity to tolerate a range of environmental stresses, such as low temperature. Cold/freezing stress affects many physiological processes in insects, causing changes in main metabolic pathways, cellular dehydration, loss of neuromuscular function, and imbalance in water and ion homeostasis. The neuroendocrine system and its related signaling mediators, such as neuropeptides and biogenic amines, play central roles in the regulation of the various physiological and behavioral processes of insects and hence can also potentially impact thermal tolerance. In response to cold stress, various chemical signals are released either via direct intercellular contact or systemically. These are signals which regulate osmoregulation - capability peptides (CAPA), inotocin (ITC)-like peptides, ion transport peptide (ITP), diuretic hormones and calcitonin (CAL), substances related to the general response to various stress factors - tachykinin-related peptides (TRPs) or peptides responsible for the mobilization of body reserves. All these processes are potentially important in cold tolerance mechanisms. This review summarizes the current knowledge on the involvement of the neuroendocrine system in the cold stress response and the possible contributions of various signaling molecules in this process.