The use of lake sedimentary DNA to track the long-term changes in both terrestrial and aquatic biota is a rapidly advancing field in paleoecological research. Although largely applied nowadays, knowledge gaps remain in this field and there is therefore still research to be conducted to ensure the reliability of the sedimentary DNA signal. Building on the most recent literature and seven original case studies, we synthesize the state-of-the-art analytical procedures for effective sampling, extraction, amplification, quantification and/or generation of DNA inventories from sedimentary ancient DNA (sedaDNA) via high-throughput sequencing technologies. We provide recommendations based on current knowledge and best practises.

Anthropogenic and climatic stressors on freshwater ecosystems are of global concern. However, the interactions and effects of multiple stressors (e.g., nutrient enrichment, climate warming, altered wind and precipitation) acting over different spatial and temporal scales are often complex and remain controversial. Here, we reconstructed one‐century dynamics of eutrophication and primary producer communities in algal‐dominated and macrophyte‐dominated zones of a large shallow lake (Taihu, China), by integrating sedimentary photosynthetic pigments and geochemical records with water monitoring and historical archives. We aimed to explore the long‐term underlying mechanisms of the responses of water quality and lake biota to multiple environmental perturbations. We found that water quality degradation and algal community modification showed similar trends but distinct timings and trajectories in contrasting ecological zones. Onset and intensity of eutrophication in north Meiliang Bay (since the 1950s) exceeded far beyond that of macrophyte‐dominated Eastern Taihu (~1990s). Anthropogenic nutrients overtook past climatic control on production and composition of phototrophic assemblages. More importantly, lake phytoplankton responded markedly to climate warming, decreasing wind speed, and extreme weathers after cultural eutrophication. Synergistic interactions of nutrients and climate on lake ecosystems became increasingly significant in promoting harmful algal blooms (HABs) dominated by Microcystis, close to the hyper‐eutrophic north lake zones. The asynchronous limnological and ecological responses also indicated the modulating roles of lake ecological regime and catchment hydrogeomorphic characteristic. Collectively, our findings suggest that mitigation of eutrophication and HABs calls for a triple management strategy integrating anthropogenic nutrients, climate change, and lake‐catchment setting.

The quantification of the abundance of aquatic organisms via the use of environmental DNA (eDNA) molecules present in water is potentially a useful tool for efficient and noninvasive population monitoring. However, questions remain about the reliability of molecular methods. Among the factors that can hamper the reliability of the eDNA quantification, we investigated the influence of five filtration methods (filter pore size, filter type) and filtered water volume (1 and 2 L) on the total eDNA and the fish eDNA concentrations of two species, brown trout (Salmo trutta) and Arctic char (Salvelinus alpinus) from tanks with known number of individuals and biomass. We applied a droplet digital PCR (ddPCR) approach to DNA extracted from water samples collected from two cultivation tanks (each of them containing one of the targeted species). Results showed that the quantification of fish eDNA concentrations of both species varies with filtration methods. More specifically, the 0.45‐µm Sterivex enclosed filters were identified to recover the highest eDNA concentrations. Difficulties to filter 2 L water samples were present for small pore size filters (≤0.45 µm) and likely caused by filter clogging. To overcome issues related to filter clogging, common in studies aiming to quantify fish eDNA molecules from water samples, we recommend a procedure involving filtration of multiple 1 L water samples with 0.45‐µm enclosed filters, to recover both high quality and high concentrations of eDNA from targeted species, and subsequent processing of independent DNA extracts with the ddPCR method.

Summary: In‐depth knowledge about spatial and temporal variation in microbial diversity and function is needed for a better understanding of ecological and evolutionary responses to global change. In particular, the study of microbial ancient DNA preserved in sediment archives from lakes and oceans can help us to evaluate the responses of aquatic microbes in the past and make predictions about future biodiversity change in those ecosystems. Recent advances in molecular genetic methods applied to the analysis of historically deposited DNA in sediments have not only allowed the taxonomic identification of past aquatic microbial communities but also enabled tracing their evolution and adaptation to episodic disturbances and gradual environmental change. Nevertheless, some challenges remain for scientists to take full advantage of the rapidly developing field of paleo‐genetics, including the limited ability to detect rare taxa and reconstruct complete genomes for evolutionary studies. Here, we provide a brief review of some of the recent advances in the field of environmental paleomicrobiology and discuss remaining challenges related to the application of molecular genetic methods to study microbial diversity, ecology, and evolution in sediment archives. We anticipate that, in the near future, environmental paleomicrobiology will shed new light on the processes of microbial genome evolution and microbial ecosystem responses to quaternary environmental changes at an unprecedented level of detail. This information can, for example, aid geological reconstructions of biogeochemical cycles and predict ecosystem responses to environmental perturbations, including in the context of human‐induced global changes.

Methylmercury (MeHg), a neurotoxic compound biomagnifying in aquatic food webs, can be a threat to human health via fish consumption. However, the composition and distribution of the microbial communities mediating the methylation of mercury (Hg) to MeHg in marine systems remain largely unknown. In order to fill this knowledge gap, we used the Baltic Sea Reference Metagenome (BARM) dataset to study the abundance and distribution of the genes involved in Hg methylation (the hgcAB gene cluster). We determined the relative abundance of the hgcAB genes and their taxonomic identity in 81 brackish metagenomes that cover spatial, seasonal and redox variability in the Baltic Sea water column. The hgcAB genes were predominantly detected in anoxic water, but some hgcAB genes were also detected in hypoxic and normoxic waters. Phylogenetic analysis identified putative Hg methylators within Deltaproteobacteria, in oxygen-deficient water layers, but also Spirochaetes-like and Kiritimatiellaeota-like bacteria. Higher relative quantities of hgcAB genes were found in metagenomes from marine particles compared to free-living communities in anoxic water, suggesting that such particles are hotspot habitats for Hg methylators in oxygen-depleted seawater. Altogether, our work unveils the diversity of the microorganisms with the potential to mediate MeHg production in the Baltic Sea and pinpoint the important ecological niches for these microorganisms within the marine water column.

Measures of environmental DNA (eDNA) concentrations in water samples have the potential to be both a cost‐efficient and a nondestructive method to estimate fish population abundance. However, the inherent temporal and spatial variability in abiotic and biotic conditions in aquatic systems have been suggested to be a major obstacle to determine relationships between fish eDNA concentrations and fish population abundance. Moreover, once water samples are collected, methodological biases are common, which introduces additional sources of variation to potential relationships between eDNA concentrations and fish population abundance. Here, we evaluate the performance of applying the droplet digital PCR (ddPCR) method to estimate fish population abundance in experimental enclosures. Using large‐scale enclosure ecosystems that contain populations of nine‐spined stickleback (Pungitius pungitius), we compared the concentrations of fish eDNA (COI mitochondrial region, 134 bp) obtained with the ddPCR method with high precision estimates of fish population abundance (i.e., number of individuals) and biomass. To evaluate the effects of contrasted concentrations of humic substances (potential PCR inhibitors) on the performance of ddPCR assays, we manipulated natural dissolved organic carbon (DOC) concentrations (range 4–11 mg/L) in the enclosures. Additionally, water temperature (+2°C) was manipulated in half of the enclosures. Results showed positive relationships between eDNA concentration and fish abundance and biomass estimates although unexplained variation remained. Still and importantly, fish eDNA estimates from high DOC enclosures were not lowered by potential inhibitory effects with our procedure. Finally, water temperature (although only 2°C difference) was neither detected as a significant factor influencing fish eDNA estimates. Altogether, our work highlights that ddPCR‐based eDNA is a promising method for future quantification of fish population abundance in natural systems.