In spite of the growing interest in the role of the gut microbiome (GM) in host physiology and health, the mechanisms governing its assembly and its effects on the environment are poorly understood. In this article, we show that the host genotype and the GM of Daphnia influence the community structure of the surrounding bacterioplankton (BPK). When Daphnia genotypes were placed in an identical environment, both the GM and BPK showed a genotype and diet-dependent taxonomic composition. Overall, the GM strongly differed from the BPK in taxonomic composition and was characterized by a lower α-diversity, suggesting a selective rejecting of bacteria from the regional species pool. In a microbiome transplant experiment, the assembly of both the GM and BPK was strongly affected by the host genotype and the inoculum to which germ-free Daphnia were exposed. The combination of these results suggests a strong interaction between the host genotype, its GM and free-living microbial communities. Currently, it is generally assumed that an animal's diet has a strong effect on the animal's GM, but only a negligible (if any) effect on the surrounding environment. However, our results indicate that the diet/microbiome inocula have a small effect on the gut community and a large effect on the community in the surrounding environment. This structuring genotype × microbiome × environment effect is an essential prerequisite that could indicate that microbiomes play an important role in eco-evolutionary processes.

The depletion of oxygen as a result of increased stratification and decreased oxygen solubility is one of the most significant chemical changes occurring in aquatic ecosystems as a result of global environmental change. Hence, more aquatic organisms will be exposed to hypoxic conditions over time. Deciphering the effects of hypoxia on strong ecological interactors in this ecosystem's food web is critical for predicting how aquatic communities can respond to such an environmental disturbance. Here, (sub-)lethal effects of hypoxia and whether these are genotype specific in Daphnia, a keystone species of freshwater ecosystems, are studied. This is especially relevant upon studying genetic responses with respect to phenotypic switches (G x E interactions) upon environmental stress. Further, we investigated the effect of hypoxia on the Daphnia microbial community to test if the microbiome plays a role in the phenotypic switch and tolerance to hypoxia. For this, two Daphnia genotypes were exposed for two weeks to either hypoxia or normoxia and host performance was monitored together with changes in the host associated and free-living microbial community after this period. We found G x E interactions for some of the tested Daphnia performance traits. The microbial community responded to hypoxia stress with responses in the bacterioplankton and in the Daphnia associated microbial community with respect to species richness and community composition and structure. The latter response was different for the two genotypes suggesting that the microbiome plays an important role in G x E interactions with respect to hypoxia tolerance in Daphnia, but further testing (e.g. through microbiome transplants) is needed to confirm this.

ABSTRACT. Human-induced increases in the bioavailability of carbon (C), nitrogen (N) and phosphorus (P) have the potential to alter the context for host-parasite dynamics in aquatic ecosystems. Given that both eutrophication and infectious diseases are becoming more prominent, it is essential to disentangle the factors that determine virulence expression in keystone grazers. Here, we focus on the impact of nutrient limitation in single versus dual parasite exposure in the water flea Daphnia magna (Crustacea, Branchiopoda). For this, we fed specimens of D. magna with algae differing in C:N:P ratios and exposed them to two virulent parasites, Pasteuria ramosa (bacteria) and the agent causing White Fat Cell (WFCD, unknown classification), both in single and dual parasite exposure treatments. Exposure to the two parasites synergistically reduced host survival, mainly driven by WFCD exposure, especially under severe nutrient limitation. Under these conditions individuals of D. magna began reproducing earlier, which resulted in a higher reproductive output upon dual parasite exposure. We here discuss these results within the framework of host stress responses, nutrient allocation and energy budgets, and conclude that the way food quality interferes with host-parasite interactions varies, depending on the parasite species involved, the nutrient requirements of all actors and the trait investigated.

In aquatic environments, interactions between cyanobacteria and their grazers are crucial for ecosystem functioning. Cyanobacteria are photosynthetic prokaryotes, which are able to produce large blooms and associated toxins, some of which are able to suppress grazer fitness. Cyanobacterial blooms are intensified by global warming and eutrophication. In our experiments, the tolerance of Daphnia magna (Straus, 1820), an efficient grazer of toxic cyanobacteria, was studied. We used different D. magna clones sampled from different sediment depths, which corresponded to different time periods of eutrophication. Our results showed that different clones had a different tolerance towards the toxic cyanobacterial species, Microcystis aeruginosa, confirming the presence of genetic variation in D. magna tolerance to cyanobacteria. However, there was not a significant adaptive effect of sediment depth. As expected, in general under controlled, infection-free conditions M. aeruginosa reduced D. magna survival. However, a coincidental, non-intended opportunistic fungal infection in a first experiment allowed us to compare the response of D. magna to M. aeruginosa in infected individuals and non-infected individuals (from a second experiment). In the presence of this opportunistic infection, there was no negative effect of M. aeruginosa in the D. magna clones, suggesting that exposure to the infection provided protection for Daphnia individuals towards Microcystis. Biotic interactions can thus be important in the interpretation of cyanobacterial effects in zooplankton grazers and in finding appropriate solutions to reduce the occurrence of cyanobacterial blooms.

Studies on stressor responses are often performed in controlled laboratory settings. The microbial communities in laboratory settings often differ from the natural environment, which could ultimately be reflected in different stress responses. In this study, we investigated the impact of single versus simultaneous multiple stressor exposure on Daphnia magna life history traits and whether this tolerance was microbiome-mediated. Daphnia individuals were exposed to the toxic cyanobacterium Microcystis aeruginosa and a fungal infection, Aspergillus aculeatus like type. Three genotypes were included to investigate genotype-specific responses. Survival, reproduction and body size were monitored for three weeks and gut microbial communities were sampled and characterized at the end of the experiment. Our study shows survival in Daphnia was microbiome-mediated as survival was only negatively impacted when Daphnia received a lab microbial community. Daphnia which received a natural microbial community have a broader environmental pool of microbiota to randomnly and selectively take up and showed no negative impact on survival. Simultaneous exposure to both stressors also revealed an antagonistic interaction for survival. Fecundity and body size were negatively impacted by exposure to stress, however, responses were here not microbiome-mediated. In addition, genotype specific responses were detected for survival and fecundity, which could be linked with the selective capabilities of the Daphnia genotypes to select beneficial or neutral microbial stains from the environment.

Studies on stressor responses are often performed in controlled laboratory settings. The microbial communities in laboratory setting often differ from the natural environment, which could ultimately be reflected in different stress responses. In this study, we investigated how stressor responses differed between laboratory and natural conditions in Daphnia magna when exposed to single or multiple stressors. Daphnia individuals were exposed to the toxic cyanobacterium Microcystis aeruginosa and a fungal infection, Aspergillus aculeatus like type. Three genotypes were included to investigate genotype-specific responses. Survival, reproduction and body size were monitored for three weeks and gut microbial communities were sampled and characterized at the end of the experiment. Our study shows that natural environments have a more diverse microbial community compared with laboratory conditions, which was ultimately reflected in the gut microbiomes after inoculation. Stressor responses in Daphnia were affected by their bacterial environment for survival, but not for fecundity and body size. Fecuntiy and body size did show a main stressor effect, which could possibly be linked with stessor-specific microbiomes (for Microcystis and the combined stressor treatment). In addition, genotype-specific responses were detected for survival and fecundity, which could be linked with the selective capabilities of the Daphnia genotypes to select beneficial or neutral microbial stains from the environment.

No abstract

The depletion of oxygen as a result of increased stratification and decreased oxygen solubility is one of the most significant chemical changes occurring in aquatic ecosystems as a result of global environmental change. Hence, more aquatic organisms will be exposed to hypoxic conditions over time. Deciphering the effects of hypoxia on strong ecological interactors in this ecosystem’s food web is critical for predicting how aquatic communities can respond to such an environmental disturbance. Here (sub-)lethal effects of hypoxia and whether these are genotype specific in Daphnia, a keystone species of freshwater ecosystems, are studied. This is especially relevant upon studying genetic responses with respect to phenotypic switches upon environmental stress. Further, we investigated the effect of hypoxia on the Daphnia microbial community to test if the microbiome plays a role in the phenotypic switch and tolerance to hypoxia. For this, two Daphnia genotypes were exposed for two weeks to either hypoxia or normoxia and host performance was monitored together with changes in the host associated and free-living microbial community after this period. We detected phenotypic plasticity for some of the tested Daphnia performance traits. The microbial community of the bacterioplankton and Daphnia associated microbial community responded via changes in species richness and community composition and structure. The latter response was different for the two genotypes suggesting that the microbiome plays an important role in phenotypic plasticity with respect to hypoxia tolerance in Daphnia, but further testing (e.g., through microbiome transplants) is needed to confirm this.