Technical challenges in metatranscriptomic studies applied to the bacterial communities of freshwater ecosystems.

Metatranscriptome analysis relates to the transcriptome of microbial communities directly sampled in the environment. Accessing the mRNA pool in natural bacterial communities presents some technical challenges such as the RNA extraction, rRNA depletion, and the choice of the high-throughput sequencing technique. The lack of technical details in scientific articles is a major problem to correctly obtained mRNA from a microbial community and thus the corresponding sequencing data. In our study, we present the methodological procedure that was developed in order to access to the metatranscriptome of the microbial communities during two cyanobacterial blooms successively occurring in a freshwater eutrophic lake. Each procedure step was detailed and discussed with regard to the choices and difficulties encountered and to the recent literature. Finally, the two major limits for metatranscriptomic approaches targeting bacterial communities from natural environments were (i) the removal of rRNA in order to increase the putative mRNA reads number after sequencing, and (ii) for most of the bacterial communities living in natural environments, the lack of reference genomes in databases that leads to the non-assignation of numerous reads. Once these challenges overcome, we managed to access putative mRNA of dominant species, i.e. cyanobacteria (from 6 to 72 % of mRNA assigned), and of the surrounding bacteria (from 1 to 5 % of mRNA assigned).

Drivers and ecological consequences of dominance in periurban phytoplankton communities using networks approaches.

Evaluating the causes and consequences of dominance by a limited number of taxa in phytoplankton communities is of huge importance in the current context of increasing anthropogenic pressures on natural ecosystems. This is of particular concern in densely populated urban areas where usages and impacts of human populations on water ecosystems are strongly interconnected. Microbial biodiversity is commonly used as a bioindicator of environmental quality and ecosystem functioning, but there are few studies at the regional scale that integrate the drivers of dominance in phytoplankton communities and their consequences on the structure and functioning of these communities. Here, we studied the causes and consequences of phytoplankton dominance in 50 environmentally contrasted waterbodies, sampled over four summer campaigns in the highly-populated Île-de-France region (IDF). Phytoplankton dominance was observed in 32-52% of the communities and most cases were attributed to Chlorophyta (35.5-40.6% of cases) and Cyanobacteria (30.3-36.5%). The best predictors of dominance were identified using multinomial logistic regression and included waterbody features (surface, depth and connection to the hydrological network) and water column characteristics (total N, TN:TP ratio, water temperature and stratification). The consequences of dominance were dependent on the identity of the dominant organisms and included modifications of biological attributes (richness, cohesion) and functioning (biomass, RUE) of phytoplankton communities. We constructed co-occurrence networks using high resolution phytoplankton biomass and demonstrated that networks under dominance by Chlorophyta and Cyanobacteria exhibited significantly different structure compared with networks without dominance. Furthermore, dominance by Cyanobacteria was associated with more profound network modifications (e.g. cohesion, size, density, efficiency and proportion of negative links), suggesting a stronger disruption of the structure and functioning of phytoplankton communities in the conditions in which this group dominates. Finally, we provide a synthesis on the relationships between environmental drivers, dominance status, community attributes and network structure.

Projecting the impact of regional land-use change and water management policies on lake water quality: an application to periurban lakes and reservoirs.

As the human population grows, the demand for living space and supplies of resources also increases, which may induce rapid change in land-use/land-cover (LULC) and associated pressures exerted on aquatic habitats. We propose a new approach to forecast the impact of regional land cover change and water management policies (i.e., targets in nutrient loads reduction) on lake and reservoir water eutrophication status using a model that requires minimal parameterisation compared with alternative methods. This approach was applied to a set of 48 periurban lakes located in the Ile de France region (IDF, France) to simulate catchment-scale management scenarios. Model outputs were subsequently compared to governmental agencies' 2030 forecasts. Our model indicated that the efforts made to reduce pressure in the catchment of seepage lakes might be expected to be proportional to the gain that might be obtained, whereas drainage lakes will display little improvement until a critical level of pressure reduction is reached. The model also indicated that remediation measures, as currently planned by governmental agencies, might only have a marginal impact on improving the eutrophication status of lakes and reservoirs within the IDF region. Despite the commitment to appropriately managing the water resources in many countries, prospective tools to evaluate the potential impacts of global change on freshwater ecosystems integrity at medium to large spatial scales are lacking. This study proposes a new approach to investigate the impact of region-scale human-driven changes on lake and reservoir ecological status and could be implemented elsewhere with limited parameterisation. Issues are discussed that relate to model uncertainty and to its relevance as a tool applied to decision-making.