In the 12 years since Dudgeon et al. (2006) reviewed major pressures on freshwater ecosystems, the biodiversity crisis in the world's lakes, reservoirs, rivers, streams and wetlands has deepened. While lakes, reservoirs and rivers cover only 2.3% of the Earth's surface, these ecosystems host at least 9.5% of the Earth's described animal species. Furthermore, using the World Wide Fund for Nature's Living Planet Index, freshwater population declines (83% between 1970 and 2014) continue to outpace contemporaneous declines in marine or terrestrial systems. The Anthropocene has brought multiple new and varied threats that disproportionately impact freshwater systems. We document 12 emerging threats to freshwater biodiversity that are either entirely new since 2006 or have since intensified: (i) changing climates; (ii) e-commerce and invasions; (iii) infectious diseases; (iv) harmful algal blooms; (v) expanding hydropower; (vi) emerging contaminants; (vii) engineered nanomaterials; (viii) microplastic pollution; (ix) light and noise; (x) freshwater salinisation; (xi) declining calcium; and (xii) cumulative stressors. Effects are evidenced for amphibians, fishes, invertebrates, microbes, plants, turtles and waterbirds, with potential for ecosystem-level changes through bottom-up and top-down processes. In our highly uncertain future, the net effects of these threats raise serious concerns for freshwater ecosystems. However, we also highlight opportunities for conservation gains as a result of novel management tools (e.g. environmental flows, environmental DNA) and specific conservation-oriented actions (e.g. dam removal, habitat protection policies, managed relocation of species) that have been met with varying levels of success. Moving forward, we advocate hybrid approaches that manage fresh waters as crucial ecosystems for human life support as well as essential hotspots of biodiversity and ecological function. Efforts to reverse global trends in freshwater degradation now depend on bridging an immense gap between the aspirations of conservation biologists and the accelerating rate of species endangerment.

Abstract. Salinization, a widespread threat to the structure and ecological functioning of inland and coastal wetlands, is currently occurring at an unprecedented rate and geographic scale. The causes of salinization are diverse and include alterations to freshwater flows, land-clearance, irrigation, disposal of wastewater effluent, sea level rise, storm surges, and applications of de-icing salts. Climate change and anthropogenic modifications to the hydrologic cycle are expected to further increase the extent and severity of wetland salinization. Salinization alters the fundamental physicochemical nature of the soil-water environment, increasing ionic concentrations and altering chemical equilibria and mineral solubility. Increased concentrations of solutes, especially sulfate, alter the biogeochemical cycling of major elements including carbon, nitrogen, phosphorus, sulfur, iron, and silica. The effects of salinization on wetland biogeochemistry typically include decreased inorganic nitrogen removal (with implications for water quality and climate regulation), decreased carbon storage (with implications for climate regulation and wetland accretion), and increased generation of toxic sulfides (with implications for nutrient cycling and the health/functioning of wetland biota). Indeed, increased salt and sulfide concentrations induce physiological stress in wetland biota and ultimately can result in large shifts in wetland communities and their associated ecosystem functions. The productivity and composition of freshwater species assemblages will be highly altered, and there is a high potential for the disruption of existing interspecific interactions. Although there is a wealth of information on how salinization impacts individual ecosystem components, relatively few studies have addressed the complex and often non-linear feedbacks that determine ecosystem-scale responses or considered how wetland salinization will affect landscape-level processes. Although the salinization of wetlands may be unavoidable in many cases, these systems may also prove to be a fertile testing ground for broader ecological theories including (but not limited to): investigations into alternative stable states and tipping points, trophic cascades, disturbance-recovery processes, and the role of historical events and landscape context in driving community response to disturbance.

Summary1. Priority question exercises are becoming an increasingly common tool to frame future agendas in conservation and ecological science. They are an effective way to identify research foci that advance the field and that also have high policy and conservation relevance. 2. To date, there has been no coherent synthesis of key questions and priority research areas for palaeoecology, which combines biological, geochemical and molecular techniques in order to reconstruct past ecological and environmental systems on time-scales from decades to millions of years. 3. We adapted a well-established methodology to identify 50 priority research questions in palaeoecology. Using a set of criteria designed to identify realistic and achievable research goals, we selected questions from a pool submitted by the international palaeoecology research community and relevant policy practitioners. 4. The integration of online participation, both before and during the workshop, increased international engagement in question selection. 5. The questions selected are structured around six themes: human-environment interactions in the Anthropocene; biodiversity, conservation and novel ecosystems; biodiversity over long time-scales; ecosystem processes and biogeochemical cycling; comparing, combining and synthesizing information from multiple records; and new developments in palaeoecology. 6. Future opportunities in palaeoecology are related to improved incorporation of uncertainty into reconstructions, an enhanced understanding of ecological and evolutionary dynamics and processes and the continued application of long-term data for better-informed landscape management. 256-26750 priority research questions in palaeoecology 257 7. Synthesis. Palaeoecology is a vibrant and thriving discipline, and these 50 priority questions highlight its potential for addressing both pure (e.g. ecological and evolutionary, methodological) and applied (e.g. environmental and conservation) issues related to ecological science and global change.

The concepts of ecosystem regime shifts, thresholds and alternative or multiple stable states are used extensively in the ecological and environmental management literature. When applied to aquatic ecosystems, these terms are used inconsistently reflecting differing levels of supporting evidence among ecosystem types. Although many aquatic ecosystems around the world have become degraded, the magnitude and causes of changes, relative to the range of historical variability, are poorly known. A working group supported by the Australian Centre for Ecological Analysis and Synthesis (ACEAS) reviewed 135 papers on freshwater ecosystems to assess the evidence for pressure-induced non-linear changes in freshwater ecosystems; these papers used terms indicating sudden and non-linear change in their titles and key words, and so was a positively biased sample. We scrutinized papers for study context and methods, ecosystem characteristics and focus, types of pressures and ecological responses considered, and the type of change reported (i.e., gradual, non-linear, hysteretic or irreversible change). There was little empirical evidence for regime shifts and changes between multiple or alternative stable states in these studies although some shifts between turbid phytoplankton-dominated states and clear-water, macrophyte-dominated states were reported in shallow lakes in temperate climates. We found limited understanding of the subtleties of the relevant theoretical concepts and encountered few mechanistic studies that investigated or identified cause-and-effect relationships between ecological responses and nominal pressures. Our results mirror those of reviews for estuarine, nearshore and marine aquatic ecosystems, demonstrating that although the concepts of regime shifts and alternative stable states have become prominent in the scientific and management literature, their empirical underpinning is weak outside of a specific environmental setting. The application of these concepts in future research and management applications should include evidence on the mechanistic links between pressures and consequent ecological change. Explicit consideration should also be given to whether observed temporal dynamics represent variation along a continuum rather than categorically different states.

The estuary of the lower River Murray features a complex mosaic of lakes, coastal lagoons and interconnecting channels. The waters of these wetlands are degraded as a result of river regulation, water abstraction, salinisation, sedimentation and the recent constriction of the River mouth. Palaeolimnologial analysis of sediment cores in two wetlands reveals that salinity in the large terminal Lake Alexandrina was only moderately influenced by tidal inflow, particularly over the past ca. 2000 years. It is now largely fresh as a result of isolation by a series of barriers completed by 1940 AD. In contrast, the seaward portion of the Coorong, a back barrier coastal lagoon, was determined to be a subsaline estuary strongly influenced by marine inflows. These findings contrast somewhat with the Coorong's current Ramsar classification as a saline lagoon. Riverine diatoms, typical of the fossil flora of Lake Alexandrina, are rare or absent in the Holocene sediments of the Coorong, other than for a short period in the late Holocene in the northernmost end of the lagoon. The palaeolimnological evidence for independent evolution of these wetlands is consistent with geomorphic evidence of a stranded, last interglacial shoreline that acted as a sill limiting the exchange of flows between Lake Alexandrina and the Coorong lagoon.

Lake sediments constitute natural archives of past environmental changes. Historically, research has focused mainly on generating regional climate records, but records of human impacts caused by land use and exploitation of freshwater resources are now attracting scientific and management interests. Long-term environmental records are useful to establish ecosystem reference conditions, enabling comparisons with current environments and potentially allowing future trajectories to be more tightly constrained. Here we review the timing and onset of human disturbance in and around inland water ecosystems as revealed through sedimentary archives from around the world. Palaeolimnology provides access to a wealth of information reflecting early human activities and their corresponding aquatic ecological shifts. First human impacts on aquatic systems and their watersheds are highly variable in time and space. Landscape disturbance often constitutes the first anthropogenic signal in palaeolimnological records. While the effects of humans at the landscape level are relatively easily demonstrated, the earliest signals of humaninduced changes in the structure and functioning of aquatic ecosystems need very careful investigation using multiple proxies. Additional studies will improve our understanding of linkages between human settlements, their exploitation of land and water resources, and the downstream effects on continental waters.

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Epipelic diatom assemblages collected from three wetlands connected to the Murray River displayed considerable variation in response to flooding and drying phases. Murray River water input usually generated diatom assemblages dominated by Aulacoseira species. After isolation, the diatom flora of two wetlands shifted to assemblages of small Fragilariaceae forms. Elevated nutrient levels corresponded with the appearance of eutraphentic taxa such as Cyclotella meneghiniana, Eolimna subminuscula, Luticola mutica and Nitzschia palea. Further evapoconcentration induced shifts to taxa tolerant of elevated salinity levels including Amphora coffeaeformis, Navicula incertata, Staurophora salina and Tryblionella hungarica. Ordination analyses reveal a strong chemical control on the diatom taxa present in the wetlands, in accordance with known ecological preferences for salinity and nutrients. The influence of nitrogen and phosphorus concentrations in controlling diatom assemblages was subordinate to salinity once conductivity values exceeded 1400 μS cm–1. The results of such biomonitoring provide a means of interpreting wetland history from fossil assemblages contained in sediment sequences.