Tools for photomotor response assay standardization in ecotoxicological studies: Example of exposure to gentamicin in the freshwater planaria Schmidtea mediterranea.

Photomotor response assay (PMR) is very useful in an ecotoxicological context because it allows evaluation of behavioral response to potential toxic compounds. However, a lack of procedure standardization makes results comparison difficult between labs and organisms. Here, we aimed to propose five different tools to standardize the PMR procedure so that it may be applied to all model species, regarding: (1) the minimum total sample size, (2) the acclimation period, (3) the number and duration of light and dark phases alternation, (4) the measured behavior, and (5) the statistical analysis. As an example of procedure application, we analyzed the effect of an exposure to the antibiotic gentamicin on the locomotion behavior during PMR in an invertebrate species: the asexual freshwater planaria Schmidtea mediterranea. We encourage future studies using PMR to follow these five tools to improve data analysis and results comparability.

Early-life exposure to permethrin affects phenotypic traits in both larval and adult mangrove rivulus Kryptolebias marmoratus.

In fishes, the impacts of environmental constraints undergone during development on the behavioural response of individuals are not well understood. Obtaining more information is important since the aquatic environment is widely exposed to pollution involving neurotoxic compounds likely to cause phenotypic changes that can affect animal fitness. We explored how early exposure to the pyrethroid insecticide permethrin (PM), a compound known for its neurotoxicity, influences the phenotypic traits in both larvae and adults of the self-fertilizing fish mangrove rivulus, Kryptolebias marmoratus. First, we investigated immediate effects of PM on larvae after one-week exposure (0-7 days post-hatching): larvae exposed to high concentration (200 µg.L-1) grew less, were less active, had negative thigmotaxis and were less likely to capture prey than control individuals and those exposed to low concentration (5 µg.L-1). No difference was found between treatments when considering oxygen consumption rate and cortisol levels. Persistent effects of early exposure to PM on adults (147-149 days post-hatching) showed that fish previously exposed to high concentration of PM overcompensated growth, leading them to finally be longer and heavier than fish from other treatments. Moreover, we evidenced that levels of cortisol interacted with early PM exposure to affect behaviours during dyadic contests. Fish were more likely to initiate fighting behaviours and were more likely to be aggressive when they have low pre-contest levels of cortisol, but these effects were less pronounced when individuals were exposed to PM. This study shows that PM can have both immediate and persistent effects on phenotypic traits in a self-fertilizing vertebrate and suggests that a pyrethroid can interact with hormones action to affect animal behaviour.

Ecdysteroids affect female reproductive status and outcome of contest over hosts in the parasitoid wasp Eupelmus vuilleti.

Ecdysteroids are a family of insect hormones that may play a role in modulating aggressive behavior in reproductive contexts. In Hymenoptera, the few studies investigating the link between ecdysteroid titers, reproduction and aggressiveness during contests concern solely eusocial species. Here, we explored whether ecdysteroid titers influenced female reproductive status as well as aggressiveness and resolution of conflict in a solitary ectoparasitoid, Eupelmus vuilleti (Hymenoptera: Eupelmidae). Eupelmus vuilleti females parasitize and feed upon juvenile stages of Callosobruchus maculatus (Coleoptera: Bruchidae). When two E. vuilleti females are simultaneously present on a patch, they tend to protect the host they exploit by displaying aggressive behaviors towards conspecific competitors. To our knowledge, nothing is known about the association between ecdysteroids and aggressiveness or the outcome of contests for host access in solitary insects. First, we quantified ecdysteroid titers that naturally circulate in females without fighting experience and after a contest over host access. Ecdysteroid titers measured after the contest did not correlate with female aggressiveness during the contest, but winner wasps had higher titers than both losers and females that did not fight. Then, we manipulated hormone titers via injection: ecdysone favored egg maturation (i.e., gonadotropic effect) within 24 h and increased almost immediately the females' probability of winning host access without affecting their aggressiveness. Our results represent an important step in understanding how hormones, such as ecdysteroids, mediate insect behavior during intraspecific competition.

The challenge hypothesis in insects.

The 'challenge hypothesis' provides a predictive framework for how the social environment influences within-species variation in hormone titers. High testosterone levels are beneficial during reproduction and competition, but they also impose costs because they may suppress traits like parental care and immunity. As a result, the challenge hypothesis predicts that individuals will change their testosterone levels to match the current social environment. Although the vast majority of work on the challenge hypothesis has focused on androgens in vertebrates, there is growing evidence that insect hormones, especially juvenile hormone (JH), may respond to social stimuli in ways that parallel androgens in vertebrates. Many insects rapidly upregulate JH titers during social competition with rivals. Some insects also modulate JH titers based on contest outcomes, with winners upregulating JH and losers downregulating JH. This review will integrate work on social modulation of hormone titers in vertebrates and insects. First, we provide background on insect hormones and describe the functional parallels between androgens and JH. Second, we review evidence that insects rapidly change JH titers in response to social competition. Finally, we highlight opportunities for future work on social modulation of hormones in insects. Overall, the challenge hypothesis provides a useful conceptual framework for hypothesis-driven research in insect endocrinology. Comparing vertebrates and insects provides insight into how selection has shaped patterns of hormone responsiveness as well as the generality of hypotheses originally developed for vertebrates.

Juvenile hormone manipulation affects female reproductive status and aggressiveness in a non-social parasitoid wasp.

In vertebrates, titers of androgens such as testosterone are known to upregulate aggressive behaviors associated with reproduction. In insects, juvenile hormone (JH) is a good candidate for studying the flexibility of insect endocrine responses because it has important effects on both reproductive processes and behavior. JH has a gonadotropic effect across a broad range of insect species, increasing ovarian development in females, and may have a role in the regulation of aggressive behavior during competition. In Hymenoptera, the functions of JH have been studied in facultatively eusocial species such as polistine wasps, bumblebees, ants and bees. Surprisingly, no work has yet focused on the relationship between JH, reproduction and aggressiveness in a non-social Hymenoptera, although it may help to understand how JH actions have evolved across taxa with different degrees of sociality. Here, we explored how JH treatment influenced: i) female reproductive status, and ii) the intensity (aggressiveness) and resolution of conflict, in Eupelmus vuilleti (Hymenoptera: Eupelmidae), a solitary ectoparasitoid wasp in which females fight over hosts. We demonstrated that intra-abdominal injections of JH increased the number of mature eggs in females after 24 h. In addition, the number of aggressive behaviors displayed by females was affected by the interaction between JH treatment and the number of mature eggs in their abdomen, but mature egg load alone predicted the outcome of staged contests. Wasps were more aggressive when they had more ready-to-lay eggs, with this effect being stronger when females were injected with JH. Moreover, females won more frequently when they had higher mature egg load. Our results highlight how JH affects egg maturation and aggressive behaviors in Eupelmus vuilleti females. To our knowledge, this is the first study showing that hormone manipulation can modulate females' reproduction status and behavior during intraspecific competition over hosts in a non-social hymenopteran parasitoid.

The 'male escape hypothesis': sex-biased metamorphosis in response to climatic drivers in a facultatively paedomorphic amphibian.

Paedomorphosis is a major evolutionary process that bypasses metamorphosis and allows reproduction in larvae. In newts and salamanders, it can be facultative with paedomorphs retaining gills and metamorphs dispersing. The evolution of these developmental processes is thought to have been driven by the costs and benefits of inhabiting aquatic versus terrestrial habitats. In this context, we aimed at testing the hypothesis that climatic drivers affect phenotypic transition and the difference across sexes because sex-ratio is biased in natural populations. Through a replicated laboratory experiment, we showed that paedomorphic palmate newts (Lissotriton helveticus) metamorphosed at a higher frequency when water availability decreased and metamorphosed earlier when temperature increased in these conditions. All responses were sex-biased, and males were more prone to change phenotype than females. Our work shows how climatic variables can affect facultative paedomorphosis and support theoretical models predicting life on land instead of in water. Moreover, because males metamorphose and leave water more often and earlier than females, these results, for the first time, give an experimental explanation for the rarity of male paedomorphosis (the 'male escape hypothesis') and suggest the importance of sex in the evolution of paedomorphosis versus metamorphosis.

Social learning in a high-risk environment: incomplete disregard for the 'minnow that cried pike' results in culturally transmitted neophobia.

Many prey species rely on conspecifics to gather information about unknown predation threats, but little is known about the role of varying environmental conditions on the efficacy of social learning. We examined predator-naive minnows that had the opportunity to learn about predators from experienced models that were raised in either a low- or high-risk environment. There were striking differences in behaviour among models; high-risk models showed a weaker response to the predator cue and became neophobic in response to the control cue (a novel odour, NO). Observers that were previously paired with low-risk models acquired a strong antipredator response only to the predator cue. However, observers that interacted with high-risk models, displayed a much weaker response to the predator odour and a weak neophobic response to the NO. This is the first study reporting such different outcomes of social learning under different environmental conditions, and suggests high-risk environments promote the cultural transmission of neophobia more so than social learning. If such a transfer can be considered similar to secondary traumatization in humans, culturally transmitted neophobia in minnows may provide a good model system for understanding more about the social ecology of fear disorders.