Transfer and bioaccumulation of pesticides in terrestrial arthropods and food webs: State of knowledge and perspectives for research.

Arthropods represent an entry point for pesticide transfers in terrestrial food webs, and pesticide accumulation in upper chain organisms, such as predators can have cascading consequences on ecosystems. However, the mechanisms driving pesticide transfer and bioaccumulation in food webs remain poorly understood. Here we review the literature on pesticide transfers mediated by terrestrial arthropods in food webs. The transfer of pesticides and their potential for bioaccumulation and biomagnification are related to the chemical properties and toxicokinetic of the substances, the resistance and detoxification abilities of the contaminated organisms, as well as by their effects on organisms' life history traits. We further identify four critical areas in which knowledge gain would improve future predictions of pesticides impacts on terrestrial food webs. First, efforts should be made regarding the effects of co-formulants and pesticides mixtures that are currently understudied. Second, progress in the sensitivity of analytical methods would allow the detection of low concentrations of pesticides in small individual arthropods. Quantifying pesticides in arthropods preys, their predators, and arthropods or vertebrates at higher trophic level would bring crucial insights into the bioaccumulation and biomagnification potential of pesticides in real-world terrestrial food webs. Finally, quantifying the influence of the trophic structure and complexity of communities on the transfer of pesticides could address several important sources of variability in bioaccumulation and biomagnification across species and food webs. This narrative review will inspire future studies aiming to quantify pesticide transfers in terrestrial food webs to better capture their ecological consequences in natural and cultivated landscapes.

Organic farming reduces pesticide load in a bird of prey.

Human activities have led to the contamination of all environmental compartments worldwide, including bird species. In birds, both the environment and maternal transfer lead to high inter-brood variability in contamination levels of pollutants, whereas intra-brood variability is generally low. However, most existing studies focused on heavy metals or persistent compounds and none, to our knowledge, addressed the variability in contamination levels of multiple pesticides and the factors influencing it. In this study, the number of pesticides detected (of 104 compounds searched) and the sum of their concentrations in the blood of 55 Montagu's harrier (Circus pygargus) nestlings from 22 nests sampled in 2021 were used as metrics of contamination levels. We investigated the effect of organic farming at the size of male's home range (i.e., 14 km2) and chicks' sex and hatching order on contamination levels. We did not find a difference between inter-brood and intra-brood variability in pesticide contamination levels, suggesting a different exposure of siblings through food items. While chicks' sex or rank did not affect their contamination level, we found that the percentage of organic farming around the nests significantly decreased the number of pesticides detected, although it did not decrease the total concentrations. This finding highlights the potential role of organic farming in reducing the exposure of birds to a pesticide cocktail.

Pesticide contamination in an intensive insect predator of honey bees.

Pesticides used for plant protection can indirectly affect target and non-target organisms and are identified as a major cause of insect decline. Depending on species interactions, pesticides can be transferred into the environment from plants to preys and predators. While the transfer of pesticides is often studied through vertebrate and aquatic exposure, arthropod predators of insects may represent valuable bioindicators of environmental exposure to pesticides. A modified QuEChERS extraction coupled with HPLC-MS/MS analysis was used to address the question of the exposure to pesticides of the invasive hornet Vespa velutina, a specialist predator of honey bees. This analytical method allows the accurate quantification of nanogram/gram levels of 42 contaminants in a sample weight that can be obtained from single individuals. Pesticide residues were analyzed in female workers from 24 different hornet nests and 13 different pesticides and 1 synergist, piperonyl butoxide, were identified and quantified. In 75 % of the explored nests, we found at least one compound and in 53 % of the positive samples we could quantify residues ranging from 0.5 to 19.5 ng.g-1. In this study, hornets from nests located in sub-urban environments were the most contaminated. Pesticide residue analysis in small and easy to collect predatory insects opens new perspectives for the study of environmental contamination and the transfer of pesticides in terrestrial trophic chains.

Neonicotinoids: Still present in farmland birds despite their ban.

Neonicotinoids (neonics) are the most widely used insecticides worldwide and are considered to be of low risk to non-target organisms such as vertebrates. Further, they are reported to be rapidly excreted and metabolized, reducing their potential toxicity. Nevertheless, growing evidence of adverse effects of neonics on farmland bird species raise questions about the purported harmless nature of these pesticides. We attempted to search for pesticide residues in species of different trophic levels and at different life stages, by using multiple bird monitoring programs on a Long-Term Socio-Ecological Research (LTSER) platform. Three passerine birds-the blackbird (Turdus merula), cirl bunting (Emberiza cirlus), and common nightingale (Luscinia megarhynchos)-that feed on seeds and invertebrates were monitored during their reproductive period, and the grey partridge (Perdix perdix) that feeds on seeds was monitored during its wintering period. We also monitored chicks of an apex predator-the Montagu's harrier (Circus pygargus)-that preys mostly upon common voles but also upon insects. We found that the birds' blood samples showed presence of residues of five neonics: three banned since 2018 in France-clothianidin, thiacloprid, and thiamethoxam-and two-dinotefuran and nitenpyram-used for veterinary purposes only. While none of these neonics was detected in blackbirds, all were present in grey partridges. Clothianidin was detected in all species, except blackbirds. Concentrations of the three banned neonics were similar or higher than concentrations found in birds monitored elsewhere before the ban. These findings raise questions about the persistence of neonics within the environment and the mode of exposure to wild fauna. Future investigations on the sublethal effects of these neonics on life-history traits of these farmland birds may help in providing a better understanding of the effects of exposure of bird populations to these insecticides, and also to the consequent effect on human health.

Pesticide impacts on avian species with special reference to farmland birds: a review.

For decades, we have observed a major biodiversity crisis impacting all taxa. Avian species have been particularly well monitored over the long term, documenting their declines. In particular, farmland birds are decreasing worldwide, but the contribution of pesticides to their decline remains controversial. Most studies addressing the effects of agrochemicals are limited to their assessment under controlled laboratory conditions, the determination of lethal dose 50 (LD50) values and testing in a few species, most belonging to Galliformes. They often ignore the high interspecies variability in sensitivity, delayed sublethal effects on the physiology, behaviour and life-history traits of individuals and their consequences at the population and community levels. Most importantly, they have entirely neglected to test for the multiple exposure pathways to which individuals are subjected in the field (cocktail effects). The present review aims to provide a comprehensive overview for ecologists, evolutionary ecologists and conservationists. We aimed to compile the literature on the effects of pesticides on bird physiology, behaviour and life-history traits, collecting evidence from model and wild species and from field and lab experiments to highlight the gaps that remain to be filled. We show how subtle nonlethal exposure might be pernicious, with major consequences for bird populations and communities. We finally propose several prospective guidelines for future studies that may be considered to meet urgent needs.

Feeding on grains containing pesticide residues is detrimental to offspring development through parental effects in grey partridge.

Numerous toxicological studies have shown that ingestion of pesticides can induce physiological stress in breeding birds, with adverse consequences on egg laying parameters and offspring quality through parental effects. However, previous studies do not mimic current levels of pesticide residues in typical landscapes, and they do not consider potential cocktail effects of pesticides as they occur in the wild. Herein, we explored whether realistic pesticide exposure affected reproduction parameters and offspring condition through parental effects in Grey partridge. We fed 24 breeding pairs with either seeds from conventional agriculture crops treated with various pesticides during cropping, or organic grains without pesticide residues as controls. The conventional and organic grain diets mimicked food options potentially encountered by wild birds in the field. The results showed that ingesting low pesticide doses over a long period had consequences on reproduction and offspring quality without altering mortality in parents or chicks. Compared with organic pairs, conventional pairs yielded smaller chicks at hatching that had a lower body mass index at 24 days old. Additionally, these chicks displayed lower haematocrit when body mass index was higher. Therefore, ingestion of conventional grains by parents resulted in chronic exposure to pesticide residues, even at low doses, and this had detrimental consequences on offspring. These results demonstrate a sublethal effect of pesticide residues through parental effects. The consequences of parental exposure on chicks might partly explain the decline in wild Grey partridge populations, which raises questions for avian conservation and demography if current agrosystem approaches are continued.

Coccidial oocyst release: once a day or all day long? Tropical bird hosts shed new light on the adaptive significance of diurnal periodicity in parasite output.

Many parasites spend part of their life cycle as infectious forms released from an infected host in the external environment, where they may encounter and infect new hosts. The emergence of infectious life stages often occurs once a day to minimize mortality in adverse environments. In bird hosts, intestinal parasites such as coccidia are generally released with feces in the late afternoon. This dynamic is adaptive since it allows avoiding desiccation and ultraviolet (UV) radiation, thus reducing mortality of oocysts in the environment until transmission to the next host. If this circadian rhythm is the result of natural selection to increase oocyst survival, we may hypothesize that oocysts will appear in feces at different times depending on the environment where hosts live. Particularly, in an environment where UV radiation and desiccation are very low, we may expect oocyst circadian release to disappear since the main selective pressure would be relaxed. We sampled different species of birds in tropical and temperate forests in spring and investigated coccidian oocyst output. A strong circadian variation in the prevalence of hosts shedding coccidian oocyst was detected for species caught in the temperate forest with an increase in prevalence in the late afternoon, whereas prevalence of birds shedding oocysts was constant over the course of the day for most species sampled in the tropical rain forest. These results are consistent with the hypothesis that oocysts' circadian output is maintained by natural selection to increase oocyst survival. We discuss the adaptive significance of diurnal periodicity in parasite output.

Do human infrastructures shape nest distribution in the landscape depending on individual personality in a farmland bird of prey?

Individuals' distribution across habitats may depend on their personality. Human activities and infrastructures are critical elements of the landscape that may impact the habitat selection process. However, depending on their personality, individuals may respond differently to these unnatural elements. In the present study, we first investigated whether some human infrastructures (buildings, roads and paths) shaped Montagu's harrier nest spatial distribution in the landscape according to female personality (boldness). Second, we tested if the reproductive success of females depended on their boldness and nest location regarding infrastructures. Using a long-term (19 years) dataset, we calculated, for each infrastructure type, the distance from each nest to the nearest infrastructure and the infrastructure density around the nest. We tested the effects of female boldness (bold vs. shy) and its interaction with egg-laying date on these six metrics. Nest location in the landscape depended on female personality and on some human infrastructures: the building density was smaller around nests from shy females than from bold ones. Nest distribution related to other infrastructure metrics did not depend on female boldness. The pattern related to building density is consistent with some habitat choice hypotheses, which are discussed. Path density around nests negatively affected reproductive success regardless of female boldness, and late breeders nested further away from paths than early breeders. Human activities on paths (more common later in the season) could lead to disturbance and a decrease in parental care, reducing reproductive success. Increasing human presence in farmlands implies a need to better understand its impact on population composition, in terms of personality. Our results suggest that individual behavioural differences should be taken into account in studies assessing the effects of human disturbance on animal populations, to propose more appropriate conservation measures.

La distribution des individus au sein de l'habitat peut dépendre de leur personnalité. Les activités et infrastructures humaines sont des éléments importants du paysage qui peuvent impacter le processus de sélection des habitats. Cependant, en fonction de leur personnalité, les individus peuvent répondre différemment à ces éléments non naturels. Dans cette étude, nous avons d'abord examiné si certaines infrastructures humaines (bâtiments, routes et chemins) influençaient la répartition spatiale des nids de busard cendré en fonction de la personnalité des femelles (témérité). Ensuite, nous avons testé si le succès reproducteur des femelles dépendait de leur témérité et de la localisation du nid par rapport aux infrastructures. En utilisant des données à long terme (19 ans), nous avons calculé, pour chaque type d'infrastructure, la distance de chaque nid à la plus proche infrastructure et la densité en infrastructures autour de chaque nid. Nous avons testé les effets de la témérité des femelles (téméraire vs. timide) et son interaction avec la date de ponte sur les six métriques. La répartition spatiale des nids dépendait de la personnalité des femelles et de certaines infrastructures humaines: la densité en bâtiments était plus faible autour des nids de femelles timides qu'autour de ceux des téméraires. La distribution des nids par rapport aux autres métriques d'infrastructure n'était pas influencée par la témérité des femelles. Le patron lié à la densité en bâtiments est en accord avec certaines hypothèses de choix d'habitat qui sont discutées. La densité en chemins autour des nids affectait négativement le succès reproducteur indépendamment de la témérité des femelles, et les femelles tardives nichaient plus loin des chemins que les précoces. Les activités humaines sur les chemins (plus fréquentes tard dans la saison de reproduction) pourraient entrainer une perturbation et une diminution des soins parentaux, réduisant ainsi le succès reproducteur. L'augmentation de la présence humaine dans les zones agricoles implique la nécessité de mieux comprendre son impact sur la composition de la population, en termes de personnalité. Nos résultats suggèrent que les différences comportementales interindividuelles devraient être prises en compte dans les études évaluant les effets des perturbations humaines sur les populations animales afin de proposer des mesures de conservations plus appropriées.

Feeding partridges with organic or conventional grain triggers cascading effects in life-history traits.

Farmland birds are declining across Europe and North America and the research of factors behind is the subject of extensive researches. Agricultural intensification is now recognized as a major factor governing the loss of biodiversity with strong evidence that pesticides induced direct bird mortality at a high dose. However, less attention has been given to the long-term effects of chronic exposure to low dose of pesticides. Here, we used an experimental procedure in which grey partridges were fed with untreated grains obtained from either organic (no pesticide) or conventional agriculture (with pesticide) for 26 weeks, thus strictly mimicking wild birds foraging on fields. We then examined a suite of life-history traits (ecophysiological and behavioural) that may ultimately, influence population dynamics. We show for the first time that ingesting low pesticide doses over a long period has long-term consequences on several major physiological pathways without inducing differential mortality. Compared to control partridges, birds exposed to chronic doses i) had less developed carotenoid-based ornaments due to lower concentrations of plasmatic carotenoids, ii) had higher activated immune system, iii) showed signs of physiological stress inducing a higher intestinal parasitic load, iv) had higher behavioural activity and body condition and v) showed lower breeding investment. Our results are consistent with a hormetic effect, in which exposure to a low dose of a chemical agent may induce a positive response, but our results also indicate that breeding adults may show impaired fitness traits bearing population consequences through reduced breeding investment or productivity. Given the current scale of use of pesticides in agrosystems, we suggest that such shifts in life-history traits may have a negative long-term impact on wild bird populations across agrosystems. We stress that long-term effects should no longer be ignored in pesticide risk assessment, where currently, only short-term effects are taken into account.

Description of long-term monitoring of farmland biodiversity in a LTSER.

Understanding the response of biodiversity to management, land use and climate change is a major challenge in farmland to halt the decline of biodiversity. Farmlands shelter a wide variety of taxa, which vary in their life cycle and habitat niches. Consequently, monitoring biodiversity from sessile annual plants to migratory birds requires dedicated protocols. In this article, we describe the protocols applied in a long-term research platform, the LTSER Zone Atelier "Plaine & Val de Sèvre" (for a full description see Bretagnolle et al. (2018) [1]). We present the data in the form of the description of monitoring protocols, which has evolved through time for arable weeds, grassland plants, ground beetles, spiders, grasshoppers, wild bees, hoverflies, butterflies, small mammals, and farmland birds (passerines, owls and various flagship species).

Activity rhythm and action range of workers of the invasive hornet predator of honeybees Vespa velutina, measured by radio frequency identification tags.

In social insects, the activity rhythm of foragers and their action range determinate the activity of the colony. In vespids, which are mostly predators, the foraging range of workers determines their maximum predation pressure round the nest. One of these species, Vespa velutina, a recently invasive species introduced into Europe, exerts a strong predation on honeybees at the hive. Therefore, the definition of its activity rhythm and spatial range of predation is of primary importance. Using radio frequency identification tags (RFID), two experiments were carried out to (a) determine their return ability (called homing) in releasing 318 individuals at different distance from their colony and (b) monitor their foraging activity rhythm and the duration of their flights based on 71 individuals followed 24 hr/24 during 2 months. The homing ability of V. velutina was evaluated to be up to 5,000 m and was not affected by the cardinal orientation of release point. The lag time to return to the nest increased with the distance of release. Most of the flight activity took place between 07:00 a.m. and 08:00 p.m., hornets doing principally short flights of less than an hour. Foraging range was thus estimated ca. 1,000 m around the nest. This study of V. velutina assisted by RFID tags provides for the first time a baseline for its potential foraging distance that increase our knowledge of this species to (a) refine more accurately models for risk assessment and (b) define security perimeter for early detection of predation on invasion front.

Insect personality: what can we learn from metamorphosis?

Ontogeny of animal personality is still an open question. Testing whether personality traits correlated with state variables (e.g. metabolic rate, hormones) and/or life history traits, and which ones are involved, requires more empirical studies. Insects with metamorphosis represent a good opportunity to tackle this question. Because of the various degrees of internal (physiological, nervous) and environmental changes linked to metamorphosis they allow testing whether these modifications drive consistency in personality traits between immature and adult stages. In this review, we establish general predictions for the effects of metamorphosis on personality in insects with complete or incomplete metamorphosis and suffering from a strong or weak niche shift after metamorphosis. We then reviewed the still rare empirical literature and discuss future research axes.

Towards sustainable and multifunctional agriculture in farmland landscapes: Lessons from the integrative approach of a French LTSER platform.

Agriculture is currently facing unprecedented challenges: ensuring food, fiber and energy production in the face of global change, maintaining the economic performance of farmers and preserving natural resources such as biodiversity and associated key ecosystem services for sustainable agriculture. Addressing these challenges requires innovative landscape scale farming systems that account for changing economic and environmental targets. These novel agricultural systems need to be recognized, accepted and promoted by all stakeholders, including local residents, and supported by public policies. Agroecosystems should be considered as socio-ecological systems and alternative farming systems should be based on ecological principles while taking societal needs into account. This requires an in-depth knowledge of the multiple interactions between sociological and ecological dynamics. Long Term Socio-Ecological Research platforms (LTSER) are ideal for acquiring this knowledge as they (i) are not constrained by traditional disciplinary boundaries, (ii) operate at a large spatial scale involving all stakeholders, and (iii) use systemic approaches to investigate biodiversity and ecosystem services. This study presents the socio-ecological research strategy from the LTSER "Zone Atelier Plaine & Val de Sèvre" (ZA PVS), a large study area where data has been sampled since 1994. Its global aim is to identify effective solutions for agricultural development and the conservation of biodiversity in farmlands. Three main objectives are targeted by the ZAPVS. The first objective is intensive monitoring of landscape features, the main taxa present and agricultural practices. The second objective is the experimental investigation, in real fields with local farmers, of important ecosystem functions and services, in relation to pesticide use, crop production and farming socio-economic value. The third aim is to involve stakeholders through participatory research, citizen science and the dissemination of scientific results. This paper underlines the relevance of LTSERs for addressing agricultural challenges, while acknowledging that there are some yet unsolved key challenges.

Personality, immune response and reproductive success: an appraisal of the pace-of-life syndrome hypothesis.

The pace-of-life syndrome (POLS) hypothesis is an extended concept of the life-history theory that includes behavioural traits. The studies challenging the POLS hypothesis often focus on the relationships between a single personality trait and a physiological and/or life-history trait. While pathogens represent a major selective pressure, few studies have been interested in testing relationships between behavioural syndrome, and several fitness components including immunity. The aim of this study was to address this question in the mealworm beetle, Tenebrio molitor, a model species in immunity studies. The personality score was estimated from a multidimensional syndrome based of four repeatable behavioural traits. In a first experiment, we investigated its relationship with two measures of fitness (reproduction and survival) and three components of the innate immunity (haemocyte concentration, and levels of activity of the phenoloxidase including the total proenzyme and the naturally activated one) to challenge the POLS hypothesis in T. molitor. Overall, we found a relationship between behavioural syndrome and reproductive success in this species, thus supporting the POLS hypothesis. We also showed a sex-specific relationship between behavioural syndrome and basal immune parameters. In a second experiment, we tested whether this observed relationship with innate immunity could be confirmed in term of differential survival after challenging by entomopathogenic bacteria, Bacillus thuringiensis. In this case, no significant relationship was evidenced. We recommend that future researchers on the POLS should control for differences in evolutionary trajectory between sexes and to pay attention to the choice of the proxy used, especially when looking at immune traits.

Vespa velutina nest distribution at a local scale: An 8-year survey of the invasive honeybee predator.

Vespa velutina is an invasive species that was observed for the first time in France and Europe in 2004, which rapidly threatened domestic honeybees with active predation. For the first time in this study, we analyzed the distribution of V. velutina nests at a local scale to understand the pattern of nesting, and in 8 years of monitoring, a total of 528 nests were surveyed. With the exception of 2 years, the nests of V. velutina were randomly distributed within the monitored area, which suggested that intraspecific competition for nesting sites and/or food was unlikely. When the data for all years were combined, an aggregated distribution of nests at the seafront in the vicinity of the oyster farm and sportfishing activities was observed. This distribution suggested that seafood, attractive to vespids, might provide a valuable alternative food source that favored colony development. We also tested the effect of spring queen trapping of V. velutina on the distribution of nests in 2014, which was the year with the greatest number of colonies within the area; the position of the traps did not influence the distribution of the nests. For the first time, this study showed that (i) intraspecific competition was unlikely to be a mechanism for population regulation, (ii) some areas were more likely favored for hornet nests, and (iii) spring queen wasp trapping was inefficient as a method to limit the distribution of the V. velutina population.

The relative abundance of hemocyte types in a polyphagous moth larva depends on diet.

Hemocytes are crucial cells of the insect immune system because of their involvement in multiple immune responses including coagulation, phagocytosis and encapsulation. There are various types of hemocytes, each having a particular role in immunity, such that variation in their relative abundance affects the outcome of the immune response. This study aims to characterize these various types of hemocytes in larvae of the grapevine pest insect Eupoecilia ambiguella, and to assess variation in their concentration as a function of larval diet and immune challenge. Four types of hemocytes were found in the hemolymph of 5th instar larvae: granulocytes, oenocytoids, plasmatocytes and spherulocytes. We found that the total concentration of hemocytes and the concentration of each hemocyte type varied among diets and in response to the immune challenge. Irrespective of the diet, the concentration of granulocytes increased following a bacterial immune challenge, while the concentration of plasmatocytes and spherulocytes differentially varied between larval diets. The concentration of oenocytoids did not vary among diets before the immune challenge but varied between larval diets in response to the challenge. These results suggest that the resistance of insect larvae to different natural enemies critically depends on the effect of larval diet on the larvae's investment into the different types of hemocytes.

Behavioral syndrome in a native and an invasive hymenoptera species.

Recent studies have focused on the role of behavior in biological invasions. Individuals may differ consistently in time for several behavioral traits (personality) which covary (behavioral syndrome) resulting in different behavioral types, some of them favoring invasion. Social hymenopterans have a strong potential to be invaders and their success depends primarily on the foundresses' ability to found viable colonies. They are expected to be active, explorative and bold for optimally establishing their nest. In Europe, 2 hornet species coexist: the native Vespa crabro and the invasive Vespa velutina. These 2 species may compete for nesting sites and we suggest that the initial success of V. velutina has been favored by its behavior in outperforming V. crabro for the traits involved in nest initiation. Here, we (i) defined the personality of V. crabro and V. velutina, (ii) tested for the existence of behavioral syndrome in these species, and (iii) compared their performances using an open-field test. Our results show that V. crabro foundresses behave consistently but not V. velutina; this lack of consistency being mainly due to reduced variance among individuals. This result questions the possibility of detecting consistent behavioral differences in species having recently undergone a strong bottleneck. Both species exhibit the same correlations between activity, boldness and exploration and V. velutina clearly outperforms V. crabro for all traits. Our results suggest that activity, boldness, and exploration are implicated in both hornet nest initiation and invasion process which contributed to explain why social hymenopterans are so successful at colonization.

Olfactory attraction of the hornet Vespa velutina to honeybee colony odors and pheromones.

Since the beginning of the last century, the number of biological invasions has continuously increased worldwide. Due to their environmental and economical consequences, invasive species are now a major concern. Social wasps are particularly efficient invaders because of their distinctive biology and behavior. Among them, the yellow-legged hornet, Vespa velutina, is a keen hunter of domestic honeybees. Its recent introduction to Europe may induce important beekeeping, pollination, and biodiversity problems. Hornets use olfactory cues for the long-range detection of food sources, in this case the location of honeybee colonies, but the exact nature of these cues remains unknown. Here, we studied the orientation behavior of V. velutina workers towards a range of hive products and protein sources, as well as towards prominent chemical substances emitted by these food sources. In a multiple choice test performed under controlled laboratory conditions, we found that hornets are strongly attracted to the odor of some hive products, especially pollen and honey. When testing specific compounds, the honeybee aggregation pheromone, geraniol, proved highly attractive. Pheromones produced by honeybee larvae or by the queen were also of interest to hornet workers, albeit to a lesser extent. Our results indicate that V. velutina workers are selectively attracted towards olfactory cues from hives (stored food, brood, and queen), which may signal a high prey density. This study opens new perspectives for understanding hornets' hunting behavior and paves the way for developing efficient trapping strategies against this invasive species.

Defensive behaviour of Apis mellifera against Vespa velutina in France: testing whether European honeybees can develop an effective collective defence against a new predator.

We investigated the prey-predator interactions between the European honeybee, Apis mellifera, and the invasive yellow-legged hornet, Vespa velutina, which first invaded France in 2004 and thereafter spread to neighbouring European countries (Spain, Portugal and Italy). Our goal was to determine how successfully honeybees are able to defend their colonies against their new predator in Europe. Experiments were conducted in the southwest of France-the point of entry of the hornet in Europe-under natural and semi-controlled field conditions. We investigated a total of eight apiaries and 95 colonies subjected to either low or high levels of predation. We analyzed hornet predatory behaviour and collective response of colonies under attack. The results showed that A. mellifera in France exhibit an inefficient and unorganized defence against V. velutina, unlike in other regions of Europe and other areas around the globe where honeybees have co-evolved with their natural Vespa predators.