DNA metabarcoding reveals metacommunity dynamics in a threatened boreal wetland wilderness.

The complexity and natural variability of ecosystems present a challenge for reliable detection of change due to anthropogenic influences. This issue is exacerbated by necessary trade-offs that reduce the quality and resolution of survey data for assessments at large scales. The Peace-Athabasca Delta (PAD) is a large inland wetland complex in northern Alberta, Canada. Despite its geographic isolation, the PAD is threatened by encroachment of oil sands mining in the Athabasca watershed and hydroelectric dams in the Peace watershed. Methods capable of reliably detecting changes in ecosystem health are needed to evaluate and manage risks. Between 2011 and 2016, aquatic macroinvertebrates were sampled across a gradient of wetland flood frequency, applying both microscope-based morphological identification and DNA metabarcoding. By using multispecies occupancy models, we demonstrate that DNA metabarcoding detected a much broader range of taxa and more taxa per sample compared to traditional morphological identification and was essential to identifying significant responses to flood and thermal regimes. We show that family-level occupancy masks high variation among genera and quantify the bias of barcoding primers on the probability of detection in a natural community. Interestingly, patterns of community assembly were nearly random, suggesting a strong role of stochasticity in the dynamics of the metacommunity. This variability seriously compromises effective monitoring at local scales but also reflects resilience to hydrological and thermal variability. Nevertheless, simulations showed the greater efficiency of metabarcoding, particularly at a finer taxonomic resolution, provided the statistical power needed to detect change at the landscape scale.

A global agenda for advancing freshwater biodiversity research.

Global freshwater biodiversity is declining dramatically, and meeting the challenges of this crisis requires bold goals and the mobilisation of substantial resources. While the reasons are varied, investments in both research and conservation of freshwater biodiversity lag far behind those in the terrestrial and marine realms. Inspired by a global consultation, we identify 15 pressing priority needs, grouped into five research areas, in an effort to support informed stewardship of freshwater biodiversity. The proposed agenda aims to advance freshwater biodiversity research globally as a critical step in improving coordinated actions towards its sustainable management and conservation.

Omics Advances in Ecotoxicology.

Toxic substances in the environment generate adverse effects at all levels of biological organization from the molecular level to community and ecosystem. Given this complexity, it is not surprising that ecotoxicologists have struggled to address the full consequences of toxic substance release at ecosystem level, due to the limits of observational and experimental tools to reveal the changes in deep structure at different levels of organization. -Omics technologies, consisting of genomics and ecogenomics, have the power to reveal, in unprecedented detail, the cellular processes of an individual or biodiversity of a community in response to environmental change with high sample/observation throughput. This represents a historic opportunity to transform the way we study toxic substances in ecosystems, through direct linkage of ecological effects with the systems biology of organisms. Three recent examples of -omics advance in the assessment of toxic substances are explored here: (1) the use of functional genomics in the discovery of novel molecular mechanisms of toxicity of chemicals in the environment; (2) the development of laboratory pipelines of dose-dependent, reduced transcriptomics to support high-throughput chemical testing at the biological pathway level; and (3) the use of eDNA metabarcoding approaches for assessing chemical effects on biological communities in mesocosm experiments and through direct observation in field monitoring. -Omics advances in ecotoxicological studies not only generate new knowledge regarding mechanisms of toxicity and environmental effect, improving the relevance and immediacy of laboratory toxicological assessment, but can provide a wholly new paradigm for ecotoxicology by linking ecological models to mechanism-based, systems biology approaches.

Shifts of community composition and population density substantially affect ecosystem function despite invariant richness.

There has been considerable focus on the impacts of environmental change on ecosystem function arising from changes in species richness. However, environmental change may affect ecosystem function without affecting richness, most notably by affecting population densities and community composition. Using a theoretical model, we find that, despite invariant richness, (1) small environmental effects may already lead to a collapse of function; (2) competitive strength may be a less important determinant of ecosystem function change than the selectivity of the environmental change driver and (3) effects on ecosystem function increase when effects on composition are larger. We also present a complementary statistical analysis of 13 data sets of phytoplankton and periphyton communities exposed to chemical stressors and show that effects on primary production under invariant richness ranged from -75% to +10%. We conclude that environmental protection goals relying on measures of richness could underestimate ecological impacts of environmental change.

Can nutrients mask community responses to insecticide mixtures?

The ecological effect of simultaneous exposure to two nutrient gradients, three insecticides and different predator intensities was investigated over a 3-week period in 80 outdoor, artificial streams using field-collected benthic invertebrates. The experimental design consisted of a 2 × 5 factorial structure with two nutrient levels (oligotrophic or mesotrophic) and five concentrations of the ternary insecticide mixture consisting of the insecticides (chlorpyrifos, dimethoate and imidacloprid). Equivalent toxic unit doses were summed to create a ternary insecticide dose (e.g., 0.1 + 0.1 + 0.1 = 0.3 TU) resulting in a range of ternary insecticide mixture toxicity (i.e., control groundwater, 0.3, 0.6, 0.9 and 1.2 TU). Two genera of insect predators, Gomphus spp. (Odonata) and Agnetina spp. (Plecoptera) were also added into each replicate stream, at densities and sizes comparable to those found at our collection site, to evaluate how the contribution of predators may change in nutrient limited (oligotrophic) versus amended (mesotrophic) systems. We describe a causal mechanism whereby the combined action of nutrients and insecticides reshaped aquatic community structure by interacting through multiple pathways. Specifically, mesotrophic conditions reduced the toxic effects of ternary insecticide mixtures for aquatic insects which, in some cases, appeared to increase abundance of aquatic insects. However, higher levels of insecticides in mesotrophic streams negated this effect and were even more toxic; for example, to aquatic insect grazers than the same insecticide doses in oligotrophic treatment levels. Effects of predators were only significant in oligotrophic streams. Evidence is provided as to how nutrient and contaminant interactions can greatly complicate the assessment of community level responses to insecticide mixtures due to direct and indirect effects of the resulting changes in the density of different genera and functional feeding groups within a community.

Biomonitoring 2.0: a new paradigm in ecosystem assessment made possible by next-generation DNA sequencing.

Biological monitoring has failed to develop from simple binary assessment outcomes of the impacted/unimpacted type, towards more diagnostic frameworks, despite significant scientific effort over the past fifty years. It is our assertion that this is largely because of the limited information content of biological samples processed by traditional morphology-based taxonomy, which is a slow, imprecise process, focused on restricted groups of organisms. We envision a new paradigm in ecosystem assessment, which we refer to as 'Biomonitoring 2.0'. This new schema employs DNA-based identification of taxa, coupled with high-throughput DNA sequencing on next-generation sequencing platforms. We discuss the transformational nature of DNA-based approaches in biodiversity discovery and ecosystem assessment and outline a path forward for their future widespread application.

Population growth rate responses of Ceriodaphnia dubia to ternary mixtures of specific acting chemicals: pharmacological versus ecotoxicological modes of action.

When considering joint toxic apical effects at higher levels of biological organization, such as the growth of populations, the so-called pharmacological mode of action that relies on toxicological mechanistic effects on molecular target sites may not be relevant. Such effects on population growth rate will depend on the extent to which juvenile and adult survival rates and production rates (juvenile developmental rates and reproduction) are affected by toxic exposure and also by the sensitivity of population growth rates to life-history changes. In such cases, the ecotoxicological mode of action, defined as the crucial life-history trait processes and/or xenobiotic-life-history trait interactions underlying a toxicological effect on population growth rate, should be considered. Life-table response experiments with the crustacean Ceriodaphnia dubia exposed to single and ternary mixtures of nine compounds were conducted to test the hypothesis that joint effects on population growth rates could be predicted from the mixture constituent ecotoxicological mode of action. Joint effects of mixtures containing pharmacologically dissimilar compounds (cadmium, λ-cyhalothrin, and chlorpyrifos) that differentially affected life-history traits contributing to population growth rates were accurately predicted by the independent-action concept. Conversely, the concentration-addition concept accurately predicted joint effects of two different mixtures: one containing pharmacologically similar acting pyrethroids that also affected similarly life-history traits, the other one that included pharmacologically dissimilar compounds (3,4-dichloroaniline, sodium bromide, and fenoxycarb) acting mainly on reproduction rates. These results indicate that when assessing combined effects on population growth rate responses, selection of mixture toxicity conceptual models based on the ecotoxicological mode of action of mixture constituents provided more accurate predictions than those based on the pharmacological mode of action.

Species traits as predictors for intrinsic sensitivity of aquatic invertebrates to the insecticide chlorpyrifos.

Ecological risk assessment (ERA) has followed a taxonomy-based approach, making the assumption that related species will show similar sensitivity to toxicants, and using safety factors or species sensitivity distributions to extrapolate from tested to untested species. In ecology it has become apparent that taxonomic approaches may have limitations for the description and understanding of species assemblages in nature. Therefore it has been proposed that the inclusion of species traits in ERA could provide a useful and alternative description of the systems under investigation. At the same time, there is a growing recognition that the use of mechanistic approaches in ERA, including conceptual and quantitative models, may improve predictive and extrapolative power. Purposefully linking traits with mechanistic effect models could add value to taxonomy-based ERA by improving our understanding of how structural and functional system facets may facilitate inter-species extrapolation. Here, we explore whether and in what ways traits can be linked purposefully to mechanistic effect models to predict intrinsic sensitivity using available data on the acute sensitivity and toxicokinetics of a range of freshwater arthropods exposed to chlorpyrifos. The results of a quantitative linking of seven different endpoints and twelve traits demonstrate that while quantitative links between traits and/or trait combinations and process based (toxicokinetic) model parameters can be established, the use of simple traits to predict classical sensitivity endpoints yields little insight. Remarkably, neither of the standard sensitivity values, i.e. the LC(50) or EC(50), showed a strong correlation with traits. Future research in this area should include a quantitative linking of toxicodynamic parameter estimations and physiological traits, and requires further consideration of how mechanistic trait-process/parameter links can be used for prediction of intrinsic sensitivity across species for different substances in ERA.

Establishing standards and assessment criteria for ecological instream flow needs in agricultural regions of Canada.

Agricultural land use can place heavy demands on regional water resources, strongly influencing the quantity and timing of water flows needed to sustain natural ecosystems. The effects of agricultural practices on streamflow conditions are multifaceted, as they also contribute to the severity of impacts arising from other stressors within the river ecosystem. Thus, river scientists need to determine the quantity of water required to sustain important aquatic ecosystem components and ecological services, to support wise apportionment of water for agricultural use. It is now apparent that arbitrarily defined minimum flows are inadequate for this task because the complex habitat requirements of the biota, which underpin the structure and function of a river ecosystem, are strongly influenced by predictable temporal variations in flow. We present an alternative framework for establishing a first-level, regional ecological instream flow needs standard based on adoption of the Indicators of Hydrologic Alteration/Range of Variability Approach as a broadly applicable hydrological assessment tool, coupling this to the Canadian Ecological Flow Index which assesses ecological responses to hydrological alteration. By explicitly incorporating a new field-based ecological assessment tool for small agricultural streams, we provide a necessary verification of altered hydrology that is broadly applicable within Canada and essential to ensure the continuous feedback between the application of flow management criteria and ecological condition.

Single versus combined lethal effects of three agricultural insecticides on larvae of the freshwater insect Chironomus dilutus.

Pesticides are currently regulated individually but are present in aquatic systems as mixtures of toxicants. In this study, the lethal effects of three agricultural insecticides--chlorpyrifos, imidacloprid and dimethoate--on Chironomus dilutus larvae were examined. Ninety-six-hour static bioassays were performed using single, binary, and ternary toxicant mixtures. The effects of the binary mixtures were investigated using the MIXTOX model to determine if the mixtures behaved additively, synergistically, or antagonistically. Dimethoate was much less toxic than chlorpyrifos or imidacloprid. Chlorpyrifos and dimethoate were not additive in mixture despite their common mode of action (MOA) and resulted in opposing effects (synergism and antagonism, respectively) when combined with imidacloprid. Synergism was observed in the ternary mixture. These results suggest that current mixture models based on MOA alone may not always be adequate in describing mixture effects, as in the current situation of mixtures with relatively few components. Other factors, such as water solubility, secondary MOA, and mechanisms of toxicity, should also be considered in future investigations of multichemical-mixture effects.

Suitability of five cladoceran species from Mexico for in situ experimentation.

Although they are highly suited for site-specific risk assessment, little attention has been given to in situ ecotoxicological experimentation in tropical regions. This study intended to assess the suitability of five freshwater cladocerans from Mexico for in situ experiments. Juveniles of Ceriodaphnia cornuta, Macrothrix triserialis group, Simocephalus vetulus, Diaphanosoma birgei, and Pseudosida cf. ramosa, from two populations, were used to perform seven day laboratory and in situ experiments. Pseudosida cf. ramosa was very sensitive to handling and was discarded from further evaluations. Ceriodaphnia cornuta reproduced significantly more neonates, with smaller coefficients of variation, than M. triserialis and S. vetulus. Also due to its worldwide distribution in the tropics, C. cornuta is suggested to be highly suitable for in situ experiments.

A new method for ranking mode-specific sensitivity of freshwater arthropods to insecticides and its relationship to biological traits.

The problem of how to deal with species sensitivity differences to toxic substances has been addressed successfully with the species sensitivity distribution (SSD), yet this has not increased understanding about the underlying mechanisms of sensitivity. Other researchers have identified the mode of action of chemicals and also biological traits of species as determinants for sensitivity, yet no systematic approach combines these factors. To achieve this, first existing data on organophosphate, carbamate, and pyrethroid toxicity and mode of action and also species trait information were mined. Second, we linked taxon sensitivity to their traits at the family level to generate empirical and mechanistic hypotheses about sensitivity-trait relationships. In this way, a mode-specific sensitivity (MSS) ranking method was developed, and tested at the taxonomic level of family and genus. The application of several quality criteria indicated overall confidence in rankings, but confidence in exact taxon rank was less certain, due to data insufficiency for certain groups. The MSS rankings were found to be applicable for trait-based approaches and were successfully linked to existing trait data to identify traits with predictive potential. Although this empirical analysis cannot test causality relationships between traits and sensitivity, testable hypotheses were generated, for further experimental investigation. Single traits as well as combinations of traits can be used to predict laboratory sensitivity to the substances tested, although associations were not as strong as in previous studies. We conclude that existing trait data are not suitable for every trait-based research question and that important traits remain to be identified and quantified in relation to the processes of toxicity, i.e., the toxicokinetics and toxicodynamics.

Riparian and in-channel habitat properties linked to dragonfly emergence.

In freshwater ecosystems, habitat alteration contributes directly to biodiversity loss. Dragonflies are sentinel species that are key invertebrate predators in both aquatic (as larvae) and terrestrial ecosystems (as adults). Understanding the habitat factors affecting dragonfly emergence can inform management practices to conserve habitats supporting these species and the functions they perform. Transitioning from larvae to adults, dragonflies leave behind larval exoskeletons (exuviae), which reveal information about the emergent population without the need for sacrificing living organisms. Capitalizing on Atlantic Canada's largest freshwater wetland, the Grand Lake Meadows (GLM) and the associated Saint John/Wolastoq River (SJWR), we studied the spatial (i.e., across the mainstem, tributary, and wetland sites) and temporal (across 3 years) variation in assemblages of emergent dragonflies (Anisoptera) and assessed the relative contribution of aquatic and terrestrial factors structuring these assemblages. The GLM complex, including the lotic SJWR and its tributaries and associated lentic wetlands, provided a range of riparian and aquatic habitat variability ideal for studying dragonfly emergence patterns across a relatively homogenous climatic region. Emergent dragonfly responses were associated with spatial, but not temporal, variation. Additionally, dragonfly communities were associated with both aquatic and terrestrial factors, while diversity was primarily associated with terrestrial factors. Specific terrestrial factors associated with the emergence of the dragonfly community included canopy cover and slope, while aquatic factors included water temperature, dissolved oxygen, and baseflow. Our results indicate that management of river habitats for dragonfly conservation should incorporate riparian habitat protection while maintaining aquatic habitat and habitat quality.

The Ecobiomics project: Advancing metagenomics assessment of soil health and freshwater quality in Canada.

Transformative advances in metagenomics are providing an unprecedented ability to characterize the enormous diversity of microorganisms and invertebrates sustaining soil health and water quality. These advances are enabling a better recognition of the ecological linkages between soil and water, and the biodiversity exchanges between these two reservoirs. They are also providing new perspectives for understanding microorganisms and invertebrates as part of interacting communities (i.e. microbiomes and zoobiomes), and considering plants, animals, and humans as holobionts comprised of their own cells as well as diverse microorganisms and invertebrates often acquired from soil and water. The Government of Canada's Genomics Research and Development Initiative (GRDI) launched the Ecobiomics Project to coordinate metagenomics capacity building across federal departments, and to apply metagenomics to better characterize microbial and invertebrate biodiversity for advancing environmental assessment, monitoring, and remediation activities. The Project has adopted standard methods for soil, water, and invertebrate sampling, collection and provenance of metadata, and nucleic acid extraction. High-throughput sequencing is located at a centralized sequencing facility. A centralized Bioinformatics Platform was established to enable a novel government-wide approach to harmonize metagenomics data collection, storage and bioinformatics analyses. Sixteen research projects were initiated under Soil Microbiome, Aquatic Microbiome, and Invertebrate Zoobiome Themes. Genomic observatories were established at long-term environmental monitoring sites for providing more comprehensive biodiversity reference points to assess environmental change.

Genetic erosion and population resilience in Daphnia longispina O.F. Müller under simulated predation and metal pressures.

This study investigated the effects of nonselective predation (culling) and metal toxicity on the genetic diversity of laboratory populations of the cladoceran Daphnia longispina. Populations, comprising five clones differing in their sensitivity to lethal concentrations of a metal-rich mine drainage effluent, were subjected to all possible combinations of three levels (absent, weak, and strong) of each stressor. Observed population densities were compared to those expected from a life history experiment, and clone frequencies were determined by allozyme profiling. Culling did not affect population density, although clonal diversity was higher than the control. Populations stressed by mine drainage recovered to their initial densities within 12 days, though the most sensitive genotypes disappeared under both weak and strong metal stress levels. Because the surviving resistant clones were shown to be the most sensitive ones to other chemicals (Cd and H+), it is suggested that successive inputs of partially lethal concentrations of different chemicals can lead to the disappearance of the population, even if the time between inputs is large enough to allow density recovery. These results suggest that changes in abundance are not enough to evaluate ecological effects of chemicals on the environment and information on co-tolerance and multiple tolerance within populations is highly valuable to prevent their extinction. Moreover, populations exposed to low levels of both stressors showed clonal diversity levels identical to controls, highlighting the importance of low level effects of nonselective stressors in the maintenance of high clonal diversity levels in Daphnia populations, and thus further supporting the intermediate disturbance hypothesis.

Cadmium toxicity and uptake by mats of the freshwater diatom: Navicula pelliculosa (Bréb) Hilse.

Contaminant uptake by algae, and its subsequent toxicity, has important ramifications for aquatic biomonitoring and environmental risk assessment. To study the effects of cadmium on diatom mats, a series of experiments was undertaken. These investigated the sensitivity of Navicula pelliculosa mats to cadmium, uptake of cadmium across a range of exposure concentrations, influence of mat biomass and thickness on uptake, and cadmium uptake by mats over time. Diatom mat formation proved to be sensitive to cadmium exposure, with a 96-h EC(50) of 31 microg/L. The rapid uptake of cadmium over 15 min was a linear function of exposure concentration and was not significantly affected by mat thickness. Cadmium uptake over time was also a linear function of exposure concentration for time periods up to 5 h. Linear uptake was likely due to the availability of algal binding sites as cadmium ions diffused through the diatom mats. Internal high-pH microenvironments may also have influenced uptake, through cadmium precipitation or enhanced adsorption within the mats. The lack of a significant relationship between mat biomass and uptake could be explained by the static water exposure conditions. Other studies have shown that cadmium uptake by algal mats was only significantly affected by biomass under flowing water conditions. Flowing water appeared to facilitate the diffusion of cadmium ions through the algal mats. Our research demonstrates the propensity of diatom mats to adsorb cadmium to achieve concentrations that could inhibit macroinvertebrate grazing. Overall, these findings contribute to a greater understanding of cadmium bioavailability in aquatic ecosystems and to the further development of benthic algae as an effective biomonitoring tool.

Watered-down biodiversity? A comparison of metabarcoding results from DNA extracted from matched water and bulk tissue biomonitoring samples.

Biomonitoring programs have evolved beyond the sole use of morphological identification to determine the composition of invertebrate species assemblages in an array of ecosystems. The application of DNA metabarcoding in freshwater systems for assessing benthic invertebrate communities is now being employed to generate biological information for environmental monitoring and assessment. A possible shift from the extraction of DNA from net-collected bulk benthic samples to its extraction directly from water samples for metabarcoding has generated considerable interest based on the assumption that taxon detectability is comparable when using either method. To test this, we studied paired water and benthos samples from a taxon-rich wetland complex, to investigate differences in the detection of arthropod taxa from each sample type. We demonstrate that metabarcoding of DNA extracted directly from water samples is a poor surrogate for DNA extracted from bulk benthic samples, focusing on key bioindicator groups. Our results continue to support the use of bulk benthic samples as a basis for metabarcoding-based biomonitoring, with nearly three times greater total richness in benthic samples compared to water samples. We also demonstrated that few arthropod taxa are shared between collection methods, with a notable lack of key bioindicator EPTO taxa in the water samples. Although species coverage in water could likely be improved through increased sample replication and/or increased sequencing depth, benthic samples remain the most representative, cost-effective method of generating aquatic compositional information via metabarcoding.

Urbanisation of floodplain ecosystems: Weight-of-evidence and network meta-analysis elucidate multiple stressor pathways.

Freshwater floodplains are dynamic, diverse ecosystems that represent important transition zones between terrestrial, riparian, subsurface and aquatic habitats. Given their historic importance in human development, floodplains have been exposed to a variety of pressures, which in combination have been instrumental in driving changes within these ecosystems. Here, we present an evidence-based framework to explore direct and indirect effects of pressures and stressors on floodplain ecosystems and test this structure within the urban landscape. Evidence was obtained from peer-reviewed scientific literature, focusing on effects of key pressures and stressors on receptors, including species composition (e.g., species presence-absence, diversity) and ecosystem function (e.g., biomass, decomposition). The strength of direct and indirect effects of individual and multiple stressors on biological receptors was quantified using two separate analyses: an evidence-weighted analysis and a quantitative network meta-analysis using data extracted from 131 studies. Results demonstrate the power of adopting a systematic framework to advance quantitative assessment of floodplain ecosystems affected by multiple stressors. While direct pathways were generally stronger and provided the core network skeleton, there were many more significant indirect pathways indicating evidence gaps in our mechanistic understanding of these processes. Indeed, the importance of indirect pathways (e.g. increase in impervious surface → increase in the accumulation rate of sediment nutrients) suggests that embracing complexity in network meta-analysis is a necessary step in revealing a more complete snapshot of the network. Results from the weight-of-evidence approach generally mirrored the direct pathway structure and demonstrated the strength of incorporating study quality alongside data sufficiency. Networks illustrated novel disturbance pathways (e.g., decrease in habitat structure → decrease in structure and function of aquatic and riparian assemblages) that can be used for hypothesis generation for future scientific enquiries. Our results highlight the broader applicability of adopting the proposed framework for assessing complex environments, such as floodplains.

Towards a general framework for the assessment of interactive effects of multiple stressors on aquatic ecosystems: Results from the Making Aquatic Ecosystems Great Again (MAEGA) workshop.

A workshop was held in Wageningen, The Netherlands, in September 2017 to collate data and literature on three aquatic ecosystem types (agricultural drainage ditches, urban floodplains, and urban estuaries), and develop a general framework for the assessment of multiple stressors on the structure and functioning of these systems. An assessment framework considering multiple stressors is crucial for our understanding of ecosystem responses within a multiply stressed environment, and to inform appropriate environmental management strategies. The framework consists of two components: (i) problem identification and (ii) impact assessment. Both assessments together proceed through the following steps: 1) ecosystem selection; 2) identification of stressors and quantification of their intensity; 3) identification of receptors or sensitive groups for each stressor; 4) identification of stressor-response relationships and their potential interactions; 5) construction of an ecological model that includes relevant functional groups and endpoints; 6) prediction of impacts of multiple stressors, 7) confirmation of these predictions with experimental and monitoring data, and 8) potential adjustment of the ecological model. Steps 7 and 8 allow the assessment to be adaptive and can be repeated until a satisfactory match between model predictions and experimental and monitoring data has been obtained. This paper is the preface of the MAEGA (Making Aquatic Ecosystems Great Again) special section that includes three associated papers which are also published in this volume, which present applications of the framework for each of the three aquatic systems.

Vulnerability of diatom communities in the Peace-Athabasca Delta to environmental change.

Habitat degradation associated with resource development is a major ecological concern, particularly in Canada's boreal zone where limited information on biodiversity is available. Habitat degradation can lead to reductions in biodiversity and ecosystem function, especially when drivers of variability and diversity patterns have not been identified for a region of interest. In this study, the distribution of diatom genera in the Peace-Athabasca Delta in northeastern Alberta was examined in relation to seasonal, geographic, and alkalinity gradients. Grab samples of six abiotic variables (total dissolved nitrogen, total dissolved phosphorus, dissolved iron, turbidity, pH, and specific conductance (SPC)) were taken from 12 remote wetlands over three sampling periods, and regressed against an ordination of diatom community composition to identify key environmental drivers of diatom community variation. Indirect gradient analysis identified two major gradients among sites. First, separation of sites among sampling periods showed successional seasonal changes in diatom community composition. Second, separation of sites from the Peace sub-delta and Birch sub-delta showed a gradient of geographic separation. Direct gradient analysis failed to explain the underlying drivers of these two gradients, but did show that alkalinity is a key driver of diatom community composition in the Embarras sub-delta, and that these sites could be particularly vulnerable to community changes associated with acidification.