Exposure to pyriproxyfen (juvenile hormone agonist) does not alter maternal care and reproduction in the European earwig.

Sublethal exposure to pesticides can alter the survival and reproduction of a wide range of non-target organisms. However, it remains unclear whether this exposure can alter behaviours that are often essential for long-term population dynamics and maintenance, such as parental care. In this study, we tested the effect of pyriproxyfen exposure (an insect growth regulator) on maternal care in the European earwig, an insect that is both used in pest control in pip-fruit orchards and considered a pest in stone fruit orchards. We exposed 424 females at doses either 10 times lower, equivalent or 10 times higher than normal application rates in French orchards. As maternal care can change over the weeks of family life, we exposed the earwig mothers at five different days before and after egg hatching. We then measured the expression of ten forms of maternal care towards eggs and juveniles, six non-caring behaviours, eggs and juvenile development, metabolic reserves in mothers at egg hatching and females' production of a terminal clutch. First, our results revealed that the three tested doses of pyriproxyfen were non-lethal and confirmed that maternal care decreased throughout both pre- and post-hatching family life. However, we did not detect any effect of pyriproxyfen on maternal care and non-care behaviours, eggs and juvenile development, quantities of lipids, proteins and glycogen in mothers at egg hatching, and on the production of a future clutch. Overall, these findings suggest that the maximal doses of pyriproxyfen authorized in French orchards is likely to have limited effects on the short- and long-term maintenance of populations of the European earwig and raises fundamental questions about the nature of the link between juvenile hormone and parental care in insects.

Earwig mothers consume the feces of their juveniles during family life.

Many animals consume the feces of their conspecifics. This allo-coprophagy can have benefits, such as access to nutrients and symbionts, but also risks for consumers, mainly due to direct contact with pathogens that develop on feces. In the European earwig Forficula auricularia, mothers and juveniles live in nests lined with their feces. This surprising habit allows juveniles to consume the feces of their siblings during family life and provides them with nutritional benefits when mothers provide low care. However, it was unclear whether earwig mothers also practice allo-coprophagy, and whether this behavior is motivated by their nutritional needs. Here, we set up four types of experimental families in which we manipulated the nutritional needs of mothers and/or juveniles and measured the effects on the production of feces by the juveniles, and the consumption of these feces by the mothers. Our results first show that fed juveniles produced more feces pellet in presence of fed compared to food-deprived mothers. We also found that, overall, about 50% of the mothers consumed juveniles feces. This consumption was both more likely and larger when the feces were produced by fed compared to food-deprived juveniles, while the proportion of feces pellets eaten was larger in food-deprived compared to fed mothers. Overall, our results reveal that allo-coprophagy involves every family member and suggest that it can have both nutritional and non-nutritional benefits for earwig mothers. Allo-coprophagy could thus favor the maintenance of mothers in the nest and, more generally, promote the early evolution of family life.

Alteration of gut microbiota with a broad-spectrum antibiotic does not impair maternal care in the European earwig.

The microbes residing within the gut of an animal host often increase their own fitness by modifying their host's physiological, reproductive and behavioural functions. Whereas recent studies suggest that they may also shape host sociality and therefore have critical effects on animal social evolution, the impact of the gut microbiota on maternal care remains unexplored. This is surprising, as this behaviour is widespread among animals, often determines the fitness of both juveniles and parents, and is essential in the evolution of complex animal societies. Here, we tested whether life-long alterations of the gut microbiota with rifampicin-a broad-spectrum antibiotic-impair pre- and post-hatching maternal care in the European earwig. Our results first confirm that rifampicin altered the mothers' gut microbial communities and indicate that the composition of the gut microbiota differs before and after egg care. Contrary to our predictions, however, the rifampicin-induced alterations of the gut microbiota did not modify pre- or post-hatching care. Independent of maternal care, rifampicin increased the females' faeces production and resulted in lighter eggs and juveniles. By contrast, rifampicin altered none of the other 21 physiological, reproductive and longevity traits measured over the 300 days of a female's lifetime. Overall, these findings reveal that altering the gut microbiota with a large spectrum antibiotic such as rifampicin does not necessarily affect host sociality. They also emphasize that not all animals have evolved a co-dependence with their microbiota and call for caution when generalizing the central role of gut microbes in host biology.

Does social thermal regulation constrain individual thermal tolerance in an ant species?

In ants, social thermal regulation is the collective maintenance of a nest temperature that is optimal for individual colony members. In the thermophilic ant Aphaenogaster iberica, two key behaviours regulate nest temperature: seasonal nest relocation and variable nest depth. Outside the nest, foragers must adapt their activity to avoid temperatures that exceed their thermal limits. It has been suggested that social thermal regulation constrains physiological and morphological thermal adaptations at the individual level. We tested this hypothesis by examining the foraging rhythms of six populations of A. iberica, which were found at different elevations (from 100 to 2,000 m) in the Sierra Nevada mountain range of southern Spain. We tested the thermal resistance of individuals from these populations under controlled conditions. Janzen's climatic variability hypothesis (CVH) states that greater climatic variability should select for organisms with broader temperature tolerances. We found that the A. iberica population at 1,300 m experienced the most extreme temperatures and that ants from this population had the highest heat tolerance (LT50 = 57.55°C). These results support CVH's validity at microclimatic scales, such as the one represented by the elevational gradient in this study. Aphaenogaster iberica maintains colony food intake levels across different elevations and mean daily temperatures by shifting its rhythm of activity. This efficient colony-level thermal regulation and the significant differences in individual heat tolerance that we observed among the populations suggest that behaviourally controlled thermal regulation does not constrain individual physiological adaptations for coping with extreme temperatures.

En hormigas, la termorregulación social es el mantenimiento colectivo de la temperatura del nido óptima para los individuos de la colonia. En la hormiga termófila Aphaenogaster iberica, hay dos comportamientos clave que regulan la temperatura del nido: la reubicación estacional y la profundidad variable del nido. Fuera del nido, las obreras recolectoras deben adaptar su actividad para evitar las temperaturas que excedan sus límites térmicos. Se ha sugerido que la termorregulación social limita las adaptaciones térmicas a nivel individual, fisiológicas y morfológicas. Examinamos esta hipótesis, estudiando los ritmos de actividad de recolección de alimento en seis poblaciones de A. iberica, a distinta altitud, desde 100 m a 2,000 m, en las montañas de Sierra Nevada, en el sur de España. Y analizamos la resistencia térmica de los individuos de estas poblaciones, en condiciones controladas. La Hipótesis de la Variabilidad Climática de Janzen (CVH) postula que una mayor variabilidad climática selecciona organismos con tolerancias térmicas más amplias. Encontramos que la población de 1,300 m era la que presentaba la mayor variabilidad climática, y que las hormigas de esta población tiene la mayor resistencia térmica individual (LT50 = 57.55°C), lo que confirma la validez de la CVH a una escala microclimática en el gradiente altitudinal estudiado. Encontramos que A. iberica puede compensar por la disminución de la temperatura media que acompaña al incremento en elevación. Las hormigas pueden cambiar sus ritmos de actividad sin afectar la entrada de alimento en la colonia, que tampoco se ve afectada por la elevación o la temperatura media diaria. A pesar de esta eficiente termorregulación a nivel de colonia, las diferencias estadísticamente significativas entre poblaciones observadas en la tolerancia térmica individual sugieren que la termorregulación controlada comportalmente no limita las adaptaciones fisiológicas individuales para enfrentarse a temperaturas extremas.

Alkaloid secretion inhibited by antibiotics in Aphaenogaster ants.

Although alkaloids are frequent in the poison glands of ants of the genus Aphaenogaster, this is not the case for A. iberica. Hypothesizing that in the genus Aphaenogaster, alkaloids are produced by symbiotic bacteria, except for A. iberica, we treated an experimental lot of both A. iberica and a 'classical' Aphaenogaster species, A. senilis, with an antibiotic. Compared to workers from a control lot, this treatment reduced considerably alkaloid production in A. senilis, whereas A. iberica did not react to the treatment. Furthermore, the treatment induced an increase in cuticular hydrocarbon quantities in A. senilis, but not in A. iberica. An analysis of the ant microbiota will be the next step to confirm our hypothesis.

The Guianese population of the fire ant Solenopsis saevissima is unicolonial.

In this study, conducted in French Guiana, a part of the native range of the fire ant Solenopsis saevissima, we compared the cuticular hydrocarbon profiles of media workers with previous results based on intraspecific aggressiveness tests. We noted a strong congruence between the two studies permitting us to delimit 2 supercolonies extending over large distances (up to 54 km), a phenomenon known as unicoloniality. Solenopsis geminata workers, taken as an out-group for cluster analyses, have a very different cuticular hydrocarbon profile. Because S. saevissima has been reported outside its native range, our conclusion is that this species has the potential to become invasive because unicoloniality (i.e., the main attribute for ants to become invasive) was shown at least for the Guianese population.

Ant cuticular response to phthalate pollution.

Phthalates are common atmospheric contaminants used in the plastic industry. Ants have been shown to constitute good bioindicators of phthalate pollution. Hence, phthalates remain trapped on ant cuticles which are mostly coated with long-chain hydrocarbons. In this study, we artificially contaminated Lasius niger ants with four phthalates: dibutyl phthalate (DBP), diisobutyl phthalate (DiBP), di(2-ethylhexyl) phthalate (DEHP), and benzyl butyl phthalate (BBP). The first three have previously been found on ants in nature in Touraine (France), while the fourth has not. The four phthalates disappeared rapidly (less than 5 days) from the cuticles of live ants. In contrast, on the cuticles of dead ants, DEHP quantities remained unchanged over time. These results indicate that phthalates are actively absorbed by the cuticles of live ants. Cuticular absorption of phthalates is nonspecific because eicosane, a nonnatural hydrocarbon on L. niger cuticle, was similarly absorbed. Ants are important ecological engineers and may serve as bioindicators of ecosystem health. We also suggest that ants and more generally terrestrial arthropods may contribute to the removal of phthalates from the local environment.

Impact of ecological doses of the most widespread phthalate on a terrestrial species, the ant Lasius niger.

Phthalates are synthetic contaminants released into the environment notably by plastic waste. Semi-volatile, they adsorb to atmospheric particles and get distributed in all ecosystems. Effects of this major anthropogenic pollution in economical species in aquatic habitats have attracted large interest. On the contrary, very few studies have focused on wild terrestrial species. Yet, these lipophilic molecules are easily trapped by insect cuticle; ants and other insects have been shown to permanently bear among their cuticular components a non-negligible proportion of phthalates, meaning that they suffer from chronic exposure to these pollutants. Oral route could also be an additional way of contamination, as phthalates tend to stick to any organic particle. We show here via a food choice experiment that Lasius niger workers can detect, and avoid feeding on, food contaminated with DEHP (DiEthyl Hexyl Phthalate), the most widespread phthalate found in nature. This suggests that the main source of contamination for ants is atmosphere and that doses measured on the cuticle correspond to the chronic exposure levels for these animals. Such an ecologically relevant dose of DEHP was used to contaminate ants in lab and to investigate their physiological impact. Over a chronic exposure (1 dose per week for 5 weeks), the egg-laying rate of queens was significantly reduced lending credence to endocrine disruptive properties of such a pollutant, as also described for aquatic invertebrates. On the contrary, short term exposure (24h) to a single dose of DEHP does not induce oxidative stress in ant workers as expected, but leads to activation of the immune system. Because of their very large distribution, their presence in virtually all terrestrial ecosystems and their representation at all trophic levels, ants could be useful indicators of contamination by phthalates, especially via monitoring the level of activation of their immune state.

Region-dependent and stage-specific effects of stress, environmental enrichment, and antidepressant treatment on hippocampal neurogenesis.

Chronic stress and depression are associated with decreased levels of hippocampal neurogenesis. On the other hand, antidepressants as well as environmental enrichment may rely in part on their pro-neurogenic effects to improve cognition and mood. Because a functional heterogeneity has been consistently reported along the septo-temporal axis of the hippocampus, regional changes in neurogenesis could differentially contribute to these effects and affect distinct hippocampal functions. Mapping these regional changes could therefore provide a better understanding of the function of newborn neurons. While some studies report region-specific effects of stress and antidepressants on neurogenesis, it is unclear whether these changes affect distinct populations of newborn neurons according to their developmental stage in a region-specific manner. By using endogenous markers and BrdU labeling we quantified the regional changes in cell proliferation and survival as well as in the number of neuronal progenitors and immature neurons following unpredictable chronic mild stress (UCMS), environmental enrichment (EE) and chronic fluoxetine (20 mg/kg/day) treatment along the septo-temporal axis of the hippocampus. EE promoted cell proliferation and survival of 4-week-old newborn cells as well as increased the number and proportion of post-mitotic immature neurons specifically within the septal hippocampus. By contrast, UCMS uniformly decreased cell proliferation, survival and immature newborn neurons but differentially affected progenitor cells with a decrease restricted to the temporal regions of the hippocampus. Whereas fluoxetine treatment in control mice affected proliferation and survival specifically in the temporal hippocampus, it reversed most of the UCMS-induced alterations all along the septo-temporal axis. These results highlight that different factors known for exerting a mood improving effect differentially regulate neurogenesis along the septo-temporal axis of the hippocampus. Such region and stage specific effects may correlate to distinct functional properties of newborn neurons along the septo-temporal axis of the hippocampus which may contribute differently to the pathophysiology of affective disorders.

Ectosymbionts and immunity in the leaf-cutting ant Acromyrmex subterraneus subterraneus.

Associations with symbiotic organisms can serve as a strategy for social insects to resist pathogens. Antibiotics produced by attine ectosymbionts (Actinobacteria) suppress the growth of Escovopsis spp., the specialized parasite of attine fungus gardens. Our objective was to evaluate whether the presence or absence of symbiotic actinobacteria covering the whole ant cuticle is related to differential immunocompetence, respiratory rate and cuticular hydrocarbons (CHs). We evaluated these parameters in three worker groups of Acromyrmex subterraneus subterraneus: External workers (EXT), internal workers with actinobacteria covering the whole body (INB) and internal workers without actinobacteria covering the whole body (INØ). We also eliminated the actinobacteria by antibiotic treatment and examined worker encapsulation response. INB ants showed lower rates of encapsulation and respiration than did the EXT and INØ ants. The lower encapsulation rate did not seem to be a cost imposed by actinomycetes because the elimination of the actinomycetes did not increase the encapsulation rate. Instead, we propose that actinobacteria confer protection to young workers until the maturation of their immune system. Actinobacteria do not seem to change nestmate recognition in these colonies. Although it is known that actinobacteria have a specific action against Escovopsis spp., our studies, along with other independent studies, indicate that actinomycetes may also be important for the individual health of the workers.

Ant cuticles: a trap for atmospheric phthalate contaminants.

Phthalates are universal contaminants. We show that they are trapped by the ant cuticles and maintained permanently at a low level, generally less than 1% of cuticular components. They are found throughout the interior of the insect, predominately in the fat body, which suggests that they are adsorbed by the cuticle. In open plastic boxes free of phthalates the ants became more contaminated with phthalates over a period of time, whereas in closed glass jars they did not. This finding suggests that the main source of pollutants is the atmosphere. Different ant species collected from multiple places showed similar levels of contamination. It appeared that in some pristine places the contamination was lower, but this needs to be confirmed. Ants can be considered as bio-indicators of phthalate pollution.

Blochmannia endosymbionts and their host, the ant Camponotus fellah: cuticular hydrocarbons and melanization.

Carpenter ants (genus Camponotus) have mutualistic, endosymbiotic bacteria of the genus Blochmannia whose main contribution to their hosts is alimentary. It was also recently demonstrated that they play a role in improving immune function as well. In this study, we show that treatment with an antibiotic produces a physiological response inducing an increase in both the quantity of cuticular hydrocarbons and in the melanization of the cuticle probably due to a nutritive and immunological deficit. We suggest that this is because it enhances the protection the cuticle provides from desiccation and also from invasions by pathogens and parasites. Nevertheless, the cuticular hydrocarbon profile is not modified by the antibiotic treatment, which indicates that nestmate recognition is not modified.

Microgynous queens in the Paleartic Ant, Manica rubida: dispersal morphs or social parasites?

In many ant species, queen size is dimorphic, with small microgynes and large macrogynes, which differ, for example, in size, insemination rate, ovary development, and dispersal tactics. These polymorphic queens often correspond with alternative reproductive strategies. The Palearctic ant, Manica rubida (Latreille) (Hymenoptera: Formicidae), lives mostly in mountainous regions in either monogynous colonies, containing one macrogynous queen or polygynous colonies, containing a few large macrogynous queens. In 1998, a colony of M. rubida was discovered containing macrogynes and many small alate microgynes that did not engage in a nuptial flight but, instead, stayed in the home nest the following winter. These microgynes were studied more closely by investigating their size, behavior, and spermatheca in relation to M. rubida macrogynes and workers. Mitochondrial DNA of macrogynes, microgynes and workers from four nests was sequenced to detect possible genetic differences between them. The microgynes were significantly smaller than the macrogynes, and the head width of the gynes was completely bimodal. The microgynes behaved like workers of the macrogynes in every experiment tested. Furthermore, the microgynes had a normal spermatheca and could be fecundated, but rarely (only one in several years). Finally, all the individuals were genetically identical, except three workers that differed by only one codon position. Because these microgynes have features of both queens and workers, their functional significance in the colony is not yet clear.

Association between repeated unpredictable chronic mild stress (UCMS) procedures with a high fat diet: a model of fluoxetine resistance in mice.

Major depressive disorder is a debilitating disease. Unfortunately, treatment with antidepressants (ADs) has limited therapeutic efficacy since resistance to AD is common. Research in this field is hampered by the lack of a reliable natural animal model of AD resistance. Depression resistance is related to various factors, including the attendance of cardiovascular risk factors and past depressive episodes. We aimed to design a rodent model of depression resistance to ADs, associating cardiovascular risk factors with repeated unpredicted chronic mild stress (UCMS). Male BALB/c mice were given either a regular (4% fat) or a high fat diet (45% fat) and subjected to two 7-week periods of UCMS separated by 6 weeks. From the second week of each UCMS procedure, vehicle or fluoxetine (10 mg/kg, i.p.) was administrated daily. The effects of the UCMS and fluoxetine in both diet conditions were assessed using physical (coat state and body weight) and behavioural tests (the reward maze test and the splash test). The results demonstrate that during the second procedure, UCMS induced behavioural changes, including coat state degradation, disturbances in self-care behaviour (splash test) and anhedonia (reward maze test) and these were reversed by fluoxetine in the regular diet condition. In contrast, the high-fat diet regimen prevented the AD fluoxetine from abolishing the UCMS-induced changes. In conclusion, by associating UCMS-an already validated animal model of depression-with high-fat diet regimen, we designed a naturalistic animal model of AD resistance related to a sub-nosographic clinical entity of depression.

Hydrocarbons in the ant Lasius niger: from the cuticle to the nest and home range marking.

The cuticular hydrocarbons (CHCs) of the ant Lasius niger are described. We observe a high local colony specificity of the body cuticular profile as predicted for a monogynous and multicolonial species. The CHCs show a low geographical variation among different locations in France. The CHCs on the legs also are colony specific, but their relative quantities are slightly different from those on the main body. For the first time, we demonstrate that the inner walls of the ant nest are coated with the same hydrocarbons as those found on the cuticle but in different proportions. The high amount of inner-nest marking and its lack of colony-specificity may explain why alien ants are not rejected once they succeed in entering the nest. The cuticular hydrocarbons also are deposited in front of the nest entrance and on the foraging arena, with a progressive increase in n-alkanes relative amounts. Chemical marks laid over the substrate are colony specific only when we consider methyl-branched alkanes. Our data confirm that these "footprint hydrocarbons" are probably deposited passively by the contact of ant tarsae with the substrate. These results suggest that the CHCs chemical profiles used by ants in colony recognition are much more complex than a single template: ants have to learn and memorize odors that vary depending on their context of perception.