Adaptive Evolution of Nearctic Deepwater Fish Vision: Implications for Assessing Functional Variation for Conservation.

Intraspecific functional variation is critical for adaptation to rapidly changing environments. For visual opsins, functional variation can be characterized in vitro and often reflects a species' ecological niche but is rarely considered in the context of intraspecific variation or the impact of recent environmental changes on species of cultural or commercial significance. Investigation of adaptation in postglacial lakes can provide key insight into how rapid environmental changes impact functional evolution. Here, we report evidence for molecular adaptation in vision in 2 lineages of Nearctic fishes that are deep lake specialists: ciscoes and deepwater sculpin. We found depth-related variation in the dim-light visual pigment rhodopsin that evolved convergently in these 2 lineages. In vitro characterization of spectral sensitivity of the convergent deepwater rhodopsin alleles revealed blue-shifts compared with other more widely distributed alleles. These blue-shifted rhodopsin alleles were only observed in deep clear postglacial lakes with underwater visual environments enriched in blue light. This provides evidence of remarkably rapid and convergent visual adaptation and intraspecific functional variation in rhodopsin. Intraspecific functional variation has important implications for conservation, and these fishes are of conservation concern and great cultural, commercial, and nutritional importance to Indigenous communities. We collaborated with the Saugeen Ojibway Nation to develop and test a metabarcoding approach that we show is efficient and accurate in recovering the ecological distribution of functionally relevant variation in rhodopsin. Our approach bridges experimental analyses of protein function and genetics-based tools used in large-scale surveys to better understand the ecological extent of adaptive functional variation.

Movement of an imperiled esocid fish in an agricultural drain.

Animal movement is increasingly affected by human alterations to habitat and climate change. In wetland systems, widespread hydrologic alterations from agriculture have changed the shape, function, and stability of shallow streams and wetland habitats. These changes in habitat quality and quantity may be especially consequential for freshwater fishes such as Grass Pickerel (Esox americanus vermiculatus), a small predatory fish found in disjunct populations across southern Ontario and listed as Special Concern under Canada's Species at Risk Act. To characterize Grass Pickerel movement response to stream-channel alterations, Fisheries and Oceans Canada implemented a tracking study to monitor the movements of a Grass Pickerel population in an agricultural drain on the Niagara Peninsula (Ontario, Canada). From 2009 to 2013, 2007 Grass Pickerel were tagged and tracked in the 37.3 km2 Beaver Creek watershed using a combination of mark-recapture surveys and eight fully automated passive integrated transponder tag antennas. Most individuals moved within 500 m (i.e., stationary fish) while 16% of the fish moved > 500 m (i.e., mobile fish), with a maximum median movement distance of 1.89 km and a maximum movement distance of 13.5 km (a long-tail distribution). Most movements occurred near the largest confluence where only a few were long-distance upstream or downstream movements. Mobile fish were larger than their stationary counterparts. Grass Pickerel in sites with higher abundance had more mobile fish, implying potential density dependence. Our results highlight that, while a long-distance dispersal ability exists in extant Grass Pickerel populations, the current conditions of riverscapes may prevent these dispersals from occurring. For declining Grass Pickerel populations, limitations to their movement ecology may substantially increase the likelihood of local extirpations.

Non-linear effects on the population performance of Bighead Carp under different maturation schedules.

Bighead Carp currently threatens to invade the Laurentian Great Lakes from the Mississippi River, but the novel climatic conditions it will encounter by expanding northwards could affect its population performance. Bighead Carp in colder climates exhibits slower growth and matures later, with later maturation typically leading to larger adult size and increased fecundity and survival. Accordingly, the life-history strategies of Bighead Carp at its northern range limits could differ from those observed in its current invaded range. To explore how population performance could differ across changing environmental conditions, we used a stage- and age-based matrix population model parameterized with values reported for Bighead Carp populations around the world. The model was used to evaluate how different ages of maturity and their resulting impacts to body size, survival, and fecundity could impact rates of population growth and establishment. Age of maturity had a non-linear effect on population growth, with maturation at intermediate ages (4-6 years) resulting in better performance. However, performance differed less between maturation ages when fecundity was allowed to increase disproportionately with body size. Greater population growth at younger ages of maturity suggest that invasion at lower latitudes could enable establishment in fewer years due to faster rates of development in warmer temperatures. Across all maturation schedules, population growth was most sensitive to the recruitment of age-1 individuals and least sensitive to adult survival, and vital rates overall varied more in their contribution to population growth at younger ages of maturity. Thus, understanding the factors that control age-1 recruitment would inform projections of population performance for Bighead Carp in the Laurentian Great Lakes. Supplementary Information: The online version contains supplementary material available at 10.1007/s10530-023-03126-z.

Ichthyoplankton metabarcoding: An efficient tool for early detection of invasive species establishment.

Detection of invasive species is critical for management but is often limited by challenges associated with capture, processing and identification of early life stages. DNA metabarcoding facilitates large-scale monitoring projects to detect establishment early. Here, we test the use of DNA metabarcoding to monitor invasive species by sequencing over 5000 fishes in bulk ichthyoplankton samples (larvae and eggs) from four rivers of ecological and cultural importance in southern Canada. We were successful in detecting species known from each river and three invasive species in two of the four rivers. This includes the first detection of early life-stage rudd in the Credit River. We evaluated whether sampling gear affected the detection of invasive species and estimates of species richness, and found that light traps outperform bongo nets in both cases. We also found that the primers used for the amplification of target sequences and the number of sequencing reads generated per sample affect the consistency of species detections. However, these factors have less impact on detections and species richness estimates than the number of samples collected and analysed. Our analyses also show that incomplete reference databases can result in incorrectly attributing DNA sequences to invasive species. Overall, we conclude that DNA metabarcoding is an efficient tool for monitoring the early establishment of invasive species by detecting evidence of reproduction but requires careful consideration of sampling design and the primers used to amplify, sequence and classify the diversity of native and potentially invasive species.

Simulating the effects of long-distance dispersal and landscape heterogeneity on the eco-evolutionary outcomes of range expansion in an invasive riverine fish, Tench (Tinca tinca).

Predicting how quickly populations expand their range and whether they will retain genetic diversity when they are introduced to new regions or track environmental conditions suited to their survival is an important applied and theoretical challenge. The literature suggests that long-distance dispersal, landscape heterogeneity and the evolution of dispersal influence populations' expansion rates and genetic diversity. We used individual-based spatially explicit simulations to examine these relationships for Tench (Tinca tinca), an invasive fish expanding its geographical range in eastern North America since the 1990s. Simulated populations varied greatly in expansion rates (1.1-28.6 patches year-1 ) and genetic diversity metrics, including changes in observed heterozygosity (-19 to +0.8%) and effective number of alleles (-0.32 to -0.01). Populations with greater dispersal distances expanded faster than those with smaller dispersal distances but exhibited considerable variation in expansion rate among local populations, implying less predictable expansions. However, they tended to retain genetic diversity as they expanded, suggesting more predictable evolutionary trajectories. In contrast, populations with smaller dispersal distances spread predictably more slowly but exhibited more variability among local populations in genetic diversity losses. Consistent with empirical data, populations spreading in a longer, narrower dispersal corridor lost more neutral genetic variation to the stochastic fixation of alleles. Given the unprecedented pace of anthropogenic environmental change and the increasing need to manage range-expanding populations, our results have conservation ramifications as they imply that the evolutionary trajectories of populations characterised by shorter dispersal distances spreading in narrower landscapes are more variable and, therefore, less predictable.

Evaluation of headstarting as a conservation tool to recover Blanding's Turtles (Emydoidea blandingii) in a highly fragmented urban landscape.

Freshwater turtle populations are declining globally as a result of anthropogenic activities. Threats to turtles in urban areas are exacerbated by road mortality and subsidized predators, which can lead to catastrophic shifts in population size and structure. Headstarting is used as a conservation tool to supplement turtle populations that may otherwise face extirpation. A headstarting program began in 2012 to recover a functionally extinct population of Blanding's Turtles (Emydoidea blandingii) 26in Rouge National Urban Park (RNUP), Ontario, Canada. The original population included five adults and one juvenile turtle. From 2014 to 2020, 270 headstarted turtles were released. The population has been monitored annually since 2014 using visual-encounter surveys, radio-telemetry, and live trapping (from 2018 onwards). We used mark-recapture and radio-telemetry data to quantify abundance, survival, and sex ratio of the headstarted turtle population. Using a Jolly-Seber model, we estimated abundance to be 183 turtles (20 turtles/ha) in 2020. Estimated survival of headstarted turtles approached 89%, except for turtles released in 2019 when survival was 43% as a result of a known mass mortality event at the study site. Pre- and post-release sex ratios were not significantly different (χ2 = 1.92; p = 0.16), but shifted from 1:1.5 to 1:1 male:female post-release. Given that the headstarted turtles have not yet reached sexual maturity, it is unclear whether headstarted turtles will reach adulthood and successfully reproduce to maintain a self-sustaining population. Thus, to evaluate the success of the headstarting program, long-term monitoring is required.

Genetic diversity and structure of a recent fish invasion: Tench (Tinca tinca) in eastern North America.

Introduced and geographically expanding populations experience similar eco-evolutionary challenges, including founder events, genetic bottlenecks, and novel environments. Theory predicts that reduced genetic diversity resulting from such phenomena limits the success of introduced populations. Using 1900 SNPs obtained from restriction-site-associated DNA sequencing, we evaluated hypotheses related to the invasion history and connectivity of an invasive population of Tench (Tinca tinca), a Eurasian freshwater fish that has been expanding geographically in eastern North America for three decades. Consistent with the reported history of a single introduction event, our findings suggest that multiple introductions from distinct genetic sources are unlikely as Tench had a small effective population size (~114 [95% CI = 106-123] individuals), no strong population subdivision across time and space, and evidence of a recent genetic bottleneck. The large genetic neighbourhood size (220 km) and weak within-population genetic substructure suggested high connectivity across the invaded range, despite the relatively large area occupied. There was some evidence for a small decay in genetic diversity as the species expanded northward, but not southward, into new habitats. As eradicating the species within a ~112 km radius would be necessary to prevent recolonization, eradicating Tench is likely not feasible at watershed-and possibly local-scales. Management should instead focus on reducing abundance in priority conservation areas to mitigate adverse impacts. Our study indicates that introduced populations can thrive and exhibit relatively high levels of genetic diversity despite severe bottlenecks (<1.5% of the ancestral effective population size) and suggests that landscape heterogeneity and population demographics can generate variability in spatial patterns of genetic diversity within a single range expansion.

Improving Widescale Monitoring of Ectoparasite Presence in Northern Canadian Wildlife with the Aid of Citizen Science.

Sampling hides from harvested animals is commonly used for passive monitoring of ectoparasites on wildlife hosts, but often relies heavily on community engagement to obtain spatially and temporally consistent samples. Surveillance of winter ticks (Dermacentor albipictus) on moose (Alces alces) and caribou (Rangifer tarandus caribou) hosts in Yukon, Canada, has relied in part on voluntary submission of hides by hunters since 2011, but few samples were submitted. To enhance sampling efforts on underrepresented moose and caribou hosts, we implemented a three-year citizen science program, the Yukon Winter Tick Monitoring Project (YWTMP), to better engage with hunters in hide sample collection. A combination of in-person and social media outreach, incentivized engagement, and standardized hide sampling kits increased voluntary submissions of moose and caribou hides almost 100-fold since surveillance began. Citizen science samples expanded the northernmost geographic extent of existing sampling efforts for moose by 480 km and for caribou by 650 km to reach 67.5° N latitude. Samples also resulted in new detections of winter ticks on moose hides that are spatially separate to those submitted for other cervids in Yukon. Findings from the YWTMP have provided an essential baseline to monitor future winter tick host-parasite dynamics in the region and highlighted priority areas for ongoing tick surveillance.

Conservation implications of revised genetic structure resulting from new population discovery: the threatened eastern sand darter (Ammocrypta pellucida) in Canada.

Human activity can put non-game fishes at higher risk of extinction because of inappropriate management action. Eastern sand darter (Ammocrypta pellucida), a small benthic fish classified as threatened across much of its northern range, inhabits increasingly fragmented sandy habitats and, as a non-game fish, may be easily overlooked in conservation efforts. In this study, the authors use genotype data from nine microsatellite loci and cytochrome oxidase I (COI) sequencing data across its northern native range to re-assess genetic structure and to characterize a newly discovered, geographically disjunct population. Previous microsatellite marker analyses had identified seven distinct population genetic clusters across the region sampled; the analysis of this study showed that the newly discovered population (West Lake, Ontario) exhibits a divergent structure. COI haplotype analysis suggests that a single haplotype recolonized the Great Lakes and surrounding water bodies after the Wisconsinan glacial period, and subsequent fluctuation in water levels and habitat fragmentation resulted in divergence of genetic clusters. Although the novel West Lake population has a common ancestral source with other populations in the broader region, its divergent genetic signature merits its consideration as a separate conservation unit. The analyses of this study highlight the potential conservation implications of the discovery of new populations, particularly those of at-risk species, even within a region that has been genetically well characterized.

The International Vertebrate Pet Trade Network and Insights from US Imports of Exotic Pets.

The international trade in exotic vertebrate pets provides key social and economic benefits but also drives associated ecological, ethical, and human health impacts. However, despite its clear importance, we currently lack a full understanding of the structure of the pet trade, hampering efforts to optimize its benefits while mitigating its negative effects. In the present article, we represent and review the structure of the pet trade as a network composed of different market actors (nodes) and trade flows (links). We identify key data gaps in this network that, if filled, would enable network analyses to pinpoint targets for management. As a case study of how data-informed networks can realize this goal, we quantified spatial and temporal patterns in pets imported to the United States. Our framework and case study illustrate how network approaches can help to inform and manage the effects of the growing demand for exotic pets.

First records of Dermacentor albipictus larvae collected by flagging in Yukon, Canada.

BACKGROUND: The winter tick (Dermacentor albipictus) has garnered significant attention throughout North America for its impact on wildlife health, and especially for moose (Alces alces), where high tick burdens may result in host hair loss, anemia, and can prove fatal. The environmental transmission of D. albipictus larvae to a host is a critical event that has direct impact on infestation success, yet in-field observations of this life stage are lacking. In Yukon, Canada, D. albipictus had previously been found on hosts, but its larval life stage had not been detected in the field, despite previous sampling attempts. METHODS: We sampled for D. albipictus larvae using traditional flagging methods in Ibex Valley and Braeburn, Yukon. Sites were sampled repeatedly for D. albipictus larvae by flagging from late August to end of October in 2018 and late August to end of November 2019. RESULTS: Larvae of D. albipictus were collected throughout Ibex Valley, at approximate densities ranging from 0.04 to 4236 larvae/100 m2. Larvae were present primarily on grassy vegetation on south-facing slopes in the Ibex Valley region and in Braeburn. Highest average larval numbers suggest peak questing activity was towards the end of September and beginning of October, as elsewhere in North America. CONCLUSIONS: To the best of our knowledge, we report the first successful collection of the off-host, larval life stage of D. albipictus by flagging, north of 60° latitude in Yukon, Canada. These new observations provide critical information on the spatial distribution of the host-seeking life stage of D. albipictus and confirm that this species is completing its whole life cycle in southern Yukon. Understanding the environmental conditions where larvae spend their vulnerable period off-host in this northern location can inform both management strategies and projections of future range expansion which may occur with a changing climate.

Functional differentiation accompanies taxonomic homogenization in freshwater fish communities.

The addition of nonnative species and loss of native species has modified the composition of communities globally. Although changes in ß-diversity have been well documented, there is a need for studies incorporating multiple time periods, more than one dimension of biodiversity, and inclusion of nestedness and turnover components to understand the underlying mechanisms structuring community composition and assembly. Here, we examined temporal changes in functional dissimilarity of fish communities of the Laurentian Great Lakes and compared these changes to those of taxonomic dissimilarity by decade from 1870 to 2010. Jaccard-derived functional dissimilarity index was used to quantify changes in functional ß-diversity within communities, between all possible pairs of communities, and using a multiple-site index among all communities. ß-diversity was partitioned into components of nestedness and turnover, and changes were examined over time. Similar to patterns in taxonomic dissimilarity, each community functionally differentiated from the historical community of 1870, with Lake Superior changing the most (~24%) and Lake Ontario the least (~14%). Although communities have become taxonomically homogenized, functional ß-diversity among all communities has increased over time, indicating functional differentiation. This is likely due to functional similarity between the communities being historically high (i.e., ~88% similar in 1870). The higher taxonomic relative to functional turnover indicates that the species being replaced between communities are functionally redundant, which could occur given the harsh environmental conditions of the region and/or as a result of the recent glacial history of the region. High functional nestedness across communities reflects dispersal limitations, with smaller communities being functional subsets of large communities closer to source populations. The functional differentiation observed is likely due to nonnative species with functional traits unique to the region establishing or the loss of functionally redundant native species; however, it is important to note that patterns of homogenization were periodically observed through time. Our study demonstrates the possible factors regulating diversity in the Laurentian Great Lakes fish communities, that patterns of taxonomic and functional ß-diversity are dynamic over time and vary in the magnitude and direction of change, and that taxonomic diversity should not be used to predict changes in functional diversity.

Seasonal variation in thermal tolerance of redside dace Clinostomus elongatus.

Organisms living in environments with oscillating temperatures may rely on plastic traits to sustain thermal tolerance during high temperature periods. Phenotypic plasticity in critical thermal maximum (CTmax) is a powerful thermoregulative strategy that enables organisms to adjust CTmax when ambient temperatures do not match thermal preference. Given that global temperatures are increasing at an unprecedented rate, identifying factors that affect the plastic response in CTmax can help predict how organisms are likely to respond to changes in their thermal landscape. Using an experimental thermal chamber in the field, we investigated the effect of short-term acclimation on the CTmax and thermal safety margin (TSM) of wild-caught redside dace, Clinostomus elongatus, (n = 197) in a northern population in Two Tree River, Ontario. Streamside CTmax trials were used to identify the maximum temperature at which redside dace maintain equilibrium, providing a powerful tool for understanding how thermal stress affects individual performance. CTmax and TSM of redside dace were sensitive to changes in temperature, regardless of season, suggesting that temperature pulses caused by climate change or urban activities can impose negative fitness consequences year round. Interestingly, an individual's recent thermal history was more influential to its thermal tolerance than the current ambient water temperature. While the CTmax of redside dace increased with body size, the effect of body size on TSM remains unclear based on our models. The results provide insight into the thermal performance of redside dace that, to date, has been difficult to assess due to the species' rarity and lack of suitable streamside protocols.

Scientists' warning on invasive alien species.

Biological invasions are a global consequence of an increasingly connected world and the rise in human population size. The numbers of invasive alien species - the subset of alien species that spread widely in areas where they are not native, affecting the environment or human livelihoods - are increasing. Synergies with other global changes are exacerbating current invasions and facilitating new ones, thereby escalating the extent and impacts of invaders. Invasions have complex and often immense long-term direct and indirect impacts. In many cases, such impacts become apparent or problematic only when invaders are well established and have large ranges. Invasive alien species break down biogeographic realms, affect native species richness and abundance, increase the risk of native species extinction, affect the genetic composition of native populations, change native animal behaviour, alter phylogenetic diversity across communities, and modify trophic networks. Many invasive alien species also change ecosystem functioning and the delivery of ecosystem services by altering nutrient and contaminant cycling, hydrology, habitat structure, and disturbance regimes. These biodiversity and ecosystem impacts are accelerating and will increase further in the future. Scientific evidence has identified policy strategies to reduce future invasions, but these strategies are often insufficiently implemented. For some nations, notably Australia and New Zealand, biosecurity has become a national priority. There have been long-term successes, such as eradication of rats and cats on increasingly large islands and biological control of weeds across continental areas. However, in many countries, invasions receive little attention. Improved international cooperation is crucial to reduce the impacts of invasive alien species on biodiversity, ecosystem services, and human livelihoods. Countries can strengthen their biosecurity regulations to implement and enforce more effective management strategies that should also address other global changes that interact with invasions.

Is Niagara Falls a barrier to gene flow in riverine fishes? A test using genome-wide SNP data from seven native species.

Since the early Holocene, fish population genetics in the Laurentian Great Lakes have been shaped by the dual influences of habitat structure and post-glacial dispersal. Riverscape genetics theory predicts that longitudinal habitat corridors and unidirectional downstream water-flow drive the downstream accumulation of genetic diversity, whereas post-glacial dispersal theory predicts that fish genetic diversity should decrease with increasing distance from glacial refugia. This study examines populations of seven native fish species codistributed above and below the 58 m high Niagara Falls - a hypothesized barrier to gene flow in aquatic species. A better understanding of Niagara Falls' role as a barrier to gene flow and dispersal is needed to identify drivers of Great Lakes genetic diversity and guide strategies to limit exotic species invasions. We used genome-wide SNPs and coalescent models to test whether populations are: (a) genetically distinct, consistent with the Niagara Falls barrier hypothesis; (b) more genetically diverse upstream, consistent with post-glacial expansion theory, or downstream, consistent with the riverscape habitat theory; and (c) have migrated either upstream or downstream past Niagara Falls. We found that genetic diversity is consistently greater below Niagara Falls and the falls are an effective barrier to migration, but two species have probably dispersed upstream past the falls after glacial retreat yet before opening of the Welland Canal. Models restricting migration to after opening of the Welland Canal were generally rejected. These results help explain how river habitat features affect aquatic species' genetic diversity and highlight the need to better understand post-glacial dispersal pathways.

Trends in the detection of aquatic non-indigenous species across global marine, estuarine and freshwater ecosystems: A 50-year perspective.

Aim: The introduction of aquatic non-indigenous species (ANS) has become a major driver for global changes in species biogeography. We examined spatial patterns and temporal trends of ANS detections since 1965 to inform conservation policy and management. Location: Global. Methods: We assembled an extensive dataset of first records of detection of ANS (1965-2015) across 49 aquatic ecosystems, including the (a) year of first collection, (b) population status and (c) potential pathway(s) of introduction. Data were analysed at global and regional levels to assess patterns of detection rate, richness and transport pathways. Results: An annual mean of 43 (±16 SD) primary detections of ANS occurred-one new detection every 8.4 days for 50 years. The global rate of detections was relatively stable during 1965-1995, but increased rapidly after this time, peaking at roughly 66 primary detections per year during 2005-2010 and then declining marginally. Detection rates were variable within and across regions through time. Arthropods, molluscs and fishes were the most frequently reported ANS. Most ANS were likely introduced as stowaways in ships' ballast water or biofouling, although direct evidence is typically absent. Main conclusions: This synthesis highlights the magnitude of recent ANS detections, yet almost certainly represents an underestimate as many ANS go unreported due to limited search effort and diminishing taxonomic expertise. Temporal rates of detection are also confounded by reporting lags, likely contributing to the lower detection rate observed in recent years. There is a critical need to implement standardized, repeated methods across regions and taxa to improve the quality of global-scale comparisons and sustain core measures over longer time-scales. It will be fundamental to fill in knowledge gaps given that invasion data representing broad regions of the world's oceans are not yet readily available and to maintain knowledge pipelines for adaptive management.

Genetic structure of endangered lake chubsucker Erimyzon sucetta in Canada reveals a differentiated population in a precarious habitat.

We used mitochondrial DNA to assess the genetic structure of endangered lake chubsucker Erimyzon sucetta across its Canadian range. We found unique mitochondrial haplotypes in Lyons Creek, a tributary of the Niagara River that faces a strong potential for habitat deterioration. Lyons Creek may therefore serve as a reservoir of unique genetic diversity. The sensitivity of Lyons Creek, combined with the genetic uniqueness of its E. sucetta population, call for further investigation into whether this population should be considered a separate designatable unit for conservation purposes.

Preadaptation and Naturalization of Nonnative Species: Darwin's Two Fundamental Insights into Species Invasion.

Predicting which nonnative species become invasive is critical for their successful management, and Charles Darwin provided predictions based on species' relatedness. However, Darwin provided two opposing predictions about the relatedness of introduced nonnatives to indigenous species. First, environmental fit is the dominant factor determining invader success; thus, we should expect that invasive species are closely related to local native residents. Alternatively, if competition is important, we should expect successful invaders are distantly related to the native residents. These opposing expectations are referred to as Darwin's naturalization conundrum. The results of studies that examine nonnative species relatedness to natives are largely inconsistent. This inconsistency arises from the fact that studies occur at different spatial and temporal scales, and at different stages of invasion, and so implicitly examine different mechanisms. Further, while species have evolved ecological differences, the mode and tempo of evolution can affect species' differences, complicating the predictions from simple hypotheses. We outline unanswered questions and provide guidelines for collecting the data required to test competing hypotheses.

Environmental DNA detection of rare and invasive fish species in two Great Lakes tributaries.

The extraction and characterization of DNA from aquatic environmental samples offers an alternative, noninvasive approach for the detection of rare species. Environmental DNA, coupled with PCR and next-generation sequencing ("metabarcoding"), has proven to be very sensitive for the detection of rare aquatic species. Our study used a custom-designed group-specific primer set and next-generation sequencing for the detection of three species at risk (Eastern Sand Darter, Ammocrypta pellucida; Northern Madtom, Noturus stigmosus; and Silver Shiner, Notropis photogenis), one invasive species (Round Goby, Neogobius melanostomus) and an additional 78 native species from two large Great Lakes tributary rivers in southern Ontario, Canada: the Grand River and the Sydenham River. Of 82 fish species detected in both rivers using capture-based and eDNA methods, our eDNA method detected 86.2% and 72.0% of the fish species in the Grand River and the Sydenham River, respectively, which included our four target species. Our analyses also identified significant positive and negative species co-occurrence patterns between our target species and other identified species. Our results demonstrate that eDNA metabarcoding that targets the fish community as well as individual species of interest provides a better understanding of factors affecting the target species spatial distribution in an ecosystem than possible with only target species data. Additionally, eDNA is easily implemented as an initial survey tool, or alongside capture-based methods, for improved mapping of species distribution patterns.

Dissecting spatiotemporal patterns of functional diversity through the lens of Darwin's naturalization conundrum.

Darwin's naturalization conundrum describes the paradigm that community assembly is regulated by two opposing processes, environmental filtering and competitive interactions, which predict both similarity and distinctiveness of species to be important for establishment. Our goal is to use long-term, large-scale, and high-resolution temporal data to examine diversity patterns over time and assess whether environmental filtering or competition plays a larger role in regulating community assembly processes. We evaluated Darwin's naturalization conundrum and how functional diversity has changed in the Laurentian Great Lakes fish community from 1870 to 2010, which has experienced frequent introductions of non-native species and extirpations of native species. We analyzed how functional diversity has changed over time by decade from 1870 to 2010 at three spatial scales (regional, lake, and habitat) to account for potential noninteractions between species at the regional and lake level. We also determined which process, environmental filtering or competitive interactions, is more important in regulating community assembly and maintenance by comparing observed patterns to what we would expect in the absence of an ecological mechanism. With the exception of one community, all analyses show that functional diversity and species richness has increased over time and that environmental filtering regulates community assembly at the regional level. When examining functional diversity at the lake and habitat level, the regulating processes become more context dependent. This study is the first to examine diversity patterns and Darwin's conundrum by integrating long-term, large-scale, and high-resolution temporal data at multiple spatial scales. Our results confirm that Darwin's conundrum is highly context dependent.