Interactions between cyanobacteria and gastropods II. Impact of toxic Planktothrix agardhii on the life-history traits of Lymnaea stagnalis.

Hepatotoxins are frequently produced by many cyanobacterial species. Microcystins (MCs) are the most frequent and widely studied hepatotoxins, with potentially hazardous repercussions on aquatic organisms. As a ubiquitous herbivore living in eutrophic freshwaters, the snail Lymnaea stagnalis (Gastropoda: Pulmonata) is particularly exposed to cyanobacteria. The toxic filamentous Planktothrix agardhii is common in temperate lakes and is therefore, a potential food resource for gastropods. In the first part of this study, we demonstrated the ingestion of toxic P. agardhii by L. stagnalis during a 5 weeks exposure, with concomitant accumulation of, on average, 60% of total MCs ingested. After 3 weeks of non-toxic food (lettuce), approximately 90% of MCs were eliminated from tissues. Here, we investigate the impact of toxic P. agardhii consumption on the life-history traits (survival, growth and fecundity), locomotion and the structure of digestive and genital glands of juvenile and adult L. stagnalis. We observed a decrease of growth regardless of age, although this was more marked in juveniles, and a reduction of fecundity in adults. Survival and locomotion were not affected. Reduction of growth and fecundity continued to be observed even after feeding of non-toxic food for 3 weeks. The structure of the digestive gland was altered during the intoxication period but not irreversibly as cells tended to recover a normal status after the 3-week detoxification period. No histopathological changes occurred in the genital gland and oocytes, and spermatozoids were present in the gonadic acini. The density of cyanobacterial suspensions used in this study was comparable to those regularly observed in lakes, particularly in eutrophic waters. These results are discussed in terms of the negative impact of toxic cyanobacteria on natural communities of freshwater gastropods, and potential cascading effects on the equilibrium and functioning of the ecosystem.

Can soil microbial diversity influence plant metabolites and life history traits of a rhizophagous insect? A demonstration in oilseed rape.

Interactions between plants and phytophagous insects play an important part in shaping the biochemical composition of plants. Reciprocally plant metabolites can influence major life history traits in these insects and largely contribute to their fitness. Plant rhizospheric microorganisms are an important biotic factor modulating plant metabolites and adaptation to stress. While plant-insects or plant-microorganisms interactions and their consequences on the plant metabolite signature are well-documented, the impact of soil microbial communities on plant defenses against phytophagous insects remains poorly known. In this study, we used oilseed rape (Brassica napus) and the cabbage root fly (Delia radicum) as biological models to tackle this question. Even though D. radicum is a belowground herbivore as a larva, its adult life history traits depend on aboveground signals. We therefore tested whether soil microbial diversity influenced emergence rate and fitness but also fly oviposition behavior, and tried to link possible effects to modifications in leaf and root metabolites. Through a removal-recolonization experiment, 3 soil microbial modalities ("high," "medium," "low") were established and assessed through amplicon sequencing of 16S and 18S ribosomal RNA genes. The "medium" modality in the rhizosphere significantly improved insect development traits. Plant-microorganism interactions were marginally associated to modulations of root metabolites profiles, which could partly explain these results. We highlighted the potential role of plant-microbial interaction in plant defenses against Delia radicum. Rhizospheric microbial communities must be taken into account when analyzing plant defenses against herbivores, being either below or aboveground.

Abdominal sensory equipment involved in external host discrimination in a solitary parasitoid wasp.

Already parasitized hosts are often of poorer quality than healthy hosts. It is therefore usually advantageous for parasitoid females to recognize and reject them. Parasitized hosts can be identified on the basis of various physical or chemical marks present on the surface or inside the hosts or their surroundings in the case of concealed host. Here we studied host discrimination behaviors of females of a certain population of Pachycrepoideus vindemmiae, a solitary ectoparasitoid, which are known to reject large-sized parasitized hosts after an abdominal examination of their surface. We first investigated females' recognition behaviors of host parasitism status when confronted to small-sized hosts (Drosophila melanogaster pupae) as host size may influence the use of different cues for host selection. We showed that, in such a situation, females also discriminate parasitized hosts after an external host exploration with the tip of their ovipositor sheath (third valvulae). We then described the sense organs present on the different parts of the ovipositor by means of scanning and transmission electron microscopy analysis. As the extremity of the third valvulae bears only one type of sensilla that appears to be chemoreceptors, we considered these sensilla as highly likely to be involved in host discrimination in P. vindemmiae. To our knowledge, this is the first time that receptors located on the ovipositor sheath are described as implicated in host discrimination in parasitoid wasps. We discuss potential chemical markers that might be detected by these receptors.

Histopathology and microcystin distribution in Lymnaea stagnalis (Gastropoda) following toxic cyanobacterial or dissolved microcystin-LR exposure.

The accumulation of hepatotoxic microcystins (MCs) in gastropods has been demonstrated to be higher following grazing of toxic cyanobacteria than from MCs dissolved in ambient water. Previous studies, however, did not adequately consider MCs covalently bound to protein phosphatases, which may represent a considerably part of the MC body burden. Thus, using an immunohistochemical method, we examined and compared the histopathology and organ distribution of covalently bound MCs in Lymnaea stagnalis following a 5-week exposure to (i) dmMC-LR, dmMC-RR, and MC-YR-producing Planktothrix agardhii (5 microg MC-LReqL(-1)) and (ii) dissolved MC-LR (33 and 100 microgL(-1)). A subsequent 3-week depuration investigated potential MC elimination and tissue regeneration. Following both exposures, bound MCs were primarily observed in the digestive gland and tract of L. stagnalis. Snails exposed to toxic cyanobacteria showed severe and widespread necrotic changes in the digestive gland co-occurring with a pronounced cytoplasmic presence of MCs in digestive cells and in the lumen of digestive lobules. Snails exposed to dissolved MC-LR showed moderate and negligible pathological changes of the digestive gland co-occurring with a restrained presence of MCs in the apical membrane of digestive cells and in the lumen of digestive lobules. These results confirm lower uptake of dissolved MC-LR and correspondingly lower cytotoxicity in the digestive gland of L. stagnalis. In contrast, after ingestion of MC-containing cyanobacterial filaments, the most likely longer residual time within the digestive gland and/or the MC variant involved (e.g., MC-YR) allowed for increased MC uptake, consequently a higher MC burden in situ and thus a more pronounced ensuing pathology. While no pathological changes were observed in kidney, foot and the genital gland, MCs were detected in spermatozoids and oocytes of all exposed snails, most likely involving a hemolymph transport from the digestive system to the genital gland. The latter results indicate the potential for adverse impact of MCs on gastropod health and reproduction as well as the possible transfer of MCs to higher trophic levels of the food web.