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Bolland, J. D.

Publications and source records attributed to Bolland, J. D..

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Pumping stations negatively affect the distribution of critically endangered European eel (Anguilla anguilla); a landscape-scale study using environmental DNA metabarcoding

Context Pumping stations pose a threat to fish globally through land use change, habitat fragmentation and entrainment risk, with the catadromous and critically endangered European eel particularly impacted. Objectives/methods Establish, model, assess and understand the present-day distribution of European eel and resident fishes in 152 pumping station catchments in a once extensive wetland (The Fens) using eDNA metabarcoding (855 samples over two and half years), with specific focus on anthropogenic influences on hydrological connectivity and habitat quality. A removal survey design maximised confidence in negative results while minimising time and consumable costs. Results Eel occurrence upstream of pumping stations was low (occupancy = 28.3%) and positively associated with catchment area, fish species richness and natural hydrological connectivity (gravity drainage or flooding) and negatively associated with distance from the tidal limit. Fish species richness replaced catchment area and improved model performance, potentially acting as a biotic indicator of habitat quality and connectivity. Pumped catchments with manually operated upstream water transfers had reduced eel presence, potentially linked to the direction of water flow or the timing of operation. By contrast, fish species richness increased in these catchments during summer, suggesting displacement into unsuitable long-term habitats. Physical habitat maintenance had no detectable effect on eel occurrence or fish species richness. Conclusions This study provides the first landscape-scale assessment of European eel distribution and drivers of occurrence in pumped river catchments. The highly novel and comprehensive insights have implications for European eel conservation as well as infrastructure and catchment management, including compliance with legislation (EC Regulation No. 1100/2007).

ecology↗

Species-specific versus community-wide assays in eDNA monitoring of European eel Anguilla anguilla: Trade-offs between detection sensitivity and the value of additional community data

Effective monitoring of the critically endangered European eel (Anguilla anguilla) is essential for conservation planning and regulatory decision-making, particularly in heavily fragmented rivers. Environmental DNA (eDNA) methods offer sensitive alternatives to traditional surveys, but there is uncertainty around whether targeted assays or community-wide approaches are better suited to achieve monitoring objectives. We compared eDNA metabarcoding and species-specific quantitative PCR (qPCR) for detecting A. anguilla across 145 pumped catchments in the Fens, East Anglia, England. All sites were sampled once initially, and sites negative for A. anguilla were re-sampled based on metabarcoding results. This allowed comparison of detection rates from a single water sample and site-level retrospective identification of sites where qPCR could have identified A. anguilla in earlier samples. The findings were also set in the context of the wider biodiversity information generated by metabarcoding. From the initial (single) water sample, qPCR detected A. anguilla at seven more sites than metabarcoding (17 versus 10). With repeated sampling, metabarcoding detected A. anguilla at 43 sites, including all but one of the sites where qPCR detected A. anguilla, and ten sites where qPCR did not detect A. anguilla within the same number of samples. Indeed, the additional sampling effort required to detect A. anguilla with metabarcoding at sites also positive with qPCR was small relative to the overall sampling effort. Furthermore, metabarcoding additionally detected 28 non-target fish species alongside fish, amphibian and mammal species of conservation concern. Our results highlight trade-offs between target-species sensitivity and the broader ecological information provided by each method, and support metabarcoding as an effective tool for a holistic conservation approach, with the additional community data outweighing the marginally increased sensitivity of qPCR.

ecology↗

Seasonal changes in fish eDNA signal vary between contrasting river types

Due to the societal reliance on goods and services provided by river systems, and their close proximity to settlements, few modern-day rivers are without significant anthropogenic modifications. The natural river hydrology is often altered as a consequence of pumping water for flood alleviation, retaining water for irrigation and modifying channels for navigation. In recent years, water pumping stations have been found to have several adverse impacts, including fish mortality (direct and indirect) and habitat fragmentation. More broadly, modern-day river systems face a myriad of anthropogenic flow and channel modifications, with varying impacts on different fish life stages. To manage such risks in line with policy, knowledge of the overall fish community and priority species present is required. It is therefore important to understand the robustness of developing survey strategies across differently managed river systems. This study investigates the seasonal patterns of environmental DNA (eDNA) metabarcoding detections from water samples, taken across three differently managed river types over a one-year period. We observed some significant seasonal variation in detection rates and fish communities; however, this variation was not consistent among river types. Despite this, we found comparatively poor fish communities upstream of pumping stations all year-round, with pumped catchments containing significantly fewer species than the adjacent main river channel and our regional control site. Finally, we highlight that seasonal variation in detectability for the overall fish community may not always reflect that of priority species. In our case, we found favourable European eel (Anguilla anguilla) detection in the summer months across all river types. It is therefore recommended that rather than focusing on overall detectability, policy driven targeted surveys should be designed with priority species ecology in mind.

ecology↗

Proving a negative; estimating species 'Confidence in Absence for Decision-Making' (CIADM) using environmental DNA monitoring

O_LIPolicy-driven decision-making is an important aspect of environmental management globally, often focused on protecting priority species. However, declining trends in freshwater biodiversity have resulted in a lack of up-to-date knowledge regarding the distribution of rare and elusive species. Furthermore, when dealing with priority species, it is sometimes more important to provide a confident assessment of absence, than merely confirm their presence. Without such confident assessments, resource intensive management plans may be misplaced, and not adequately targeted to conserve important remaining populations. C_LIO_LIHere, we present a framework to estimate confidence in absence, referred to as "Confidence in Absence for Decision-Making" (CIADM), based on single-visit environmental DNA metabarcoding data obtained from water samples. It uses a case study of European eel presence / absence upstream of 44 water pumping stations, given their critically endangered status and the legislative drivers for remediation (EC Eel Regulation 1100/2007, Eels (England and Wales) Regulation 2009). Through a high degree of biological (sample) and technical (PCR) replication, we retrospectively assigned confidence in absence values and proposed various strategies to achieve the required confidence levels in future surveys. C_LIO_LIOur findings indicate that 17 out of 44 pumping stations tested positive for eel, and we were able to assign a >99% confidence level that the remaining 27 sites were negative for eel DNA at the time of sampling. Increasing both biological and technical replication increased confidence in absence values. For example, using three PCR replicates per sample, required four replicate biological samples to achieve >95% and six to achieve >99% confidence in eel absence given non-detection. However, we estimate that by using seven PCR replicates per sample a >99% confidence in eel absence following non-detection could be achieved with only three replicate biological samples. Furthermore, we found that eel positive sites had significantly higher species richness, and fish communities differed between eel positive and eel negative sites. C_LIO_LIThis study highlights the importance of optimising workflow specific replication, and provides an adaptable framework to produce confidence estimations of priority species absence given non-detection. C_LI

ecology↗

Spatial ecology and population dynamics of brown trout Salmo trutta L. in reservoirs and headwater tributaries

This investigation compared the spatial ecology and population dynamics of brown trout Salmo trutta L. between reservoirs with (impact; Langsett Reservoir) and without (control; Grimwith Reservoir) barriers to fish movements into afferent headwater tributaries, including the effectiveness of a fish pass to remediate connectivity. Passive Integrated Transponder (PIT) telemetry revealed fish that emigrated from Langsett and Grimwith reservoirs were 1-3 and 0-2 years old, respectively, and predominantly did so in March-May and October-December in both. Weirs at Langsett Reservoir (emigration rate = 26%) appeared to thwart emigration relative to Grimwith Reservoir (emigration rate = 85%). Acoustic telemetry (2D positions) in the impact reservoir revealed the largest home range was in October - December (monthly K95 {+/-} S.D. up to 26.9 {+/-} 6.69 ha in November), activity was influenced by both month and time of day, and fish occupied shallow water depths (relative to reservoir depth), especially at night. Large proportions of brown trout tagged in Grimwith and Langsett reservoirs (42.9% and 64.1%, respectively) and fish that emigrated (37.2% and 27.7%, respectively) were detected moving upstream into tributaries. At both reservoirs, peak immigration for 3- and [≥]4-year-old fish occurred in October-December, although upstream movements occurred throughout the year and by all age classes. Three brown trout passed upstream of each of the weirs on River Little Don (prior to fish pass construction; 3% of those that approached from downstream) and Thickwoods Brook (throughout the study; 2%). Overall fish pass solution passage efficiency was 14% but was higher for 2- and 3-year-old fish (32%), which was comparable to fish translocated from upstream (33%). Passage predominantly occurred at lower river levels than fish pass entrance / attraction, which was also lower than during approaches to the weir. A Before-After Control-Impact (BACI) design found that although juvenile (0+, but not >0+) brown trout densities were lower after fish pass construction, the reduction was significantly less than at control sites, i.e., the fish pass had a positive effect. Overall, this investigation significantly furthers our understanding of brown trout spatial ecology and population dynamics in reservoirs and headwater tributaries.

ecology↗

Individual genotype but not phenotype predicts river migration success in Atlantic salmon

Migratory species typically undertake demanding long-distance journeys, across different habitat types during which they are exposed to multiple natural and anthropogenic stressors. Mortality during migration is typically high, and may be exacerbated by human-induced pressures. Understanding individual responses to these selection pressures is rarely attempted, because of the challenges of relating individual phenotypic and genetic data to migration success. Here we show distinct Single Nucleotide Polymorphism (SNP) sets significantly differentiated between Atlantic salmon smolts making successful migrations to sea and those that failed to migrate, in two different rivers. In contrast, morphological variation was not diagnostic of migration success. Populations from each river were genetically distinct, and while different genes were possibly implicated in migration success in each river, they related to common biological processes (for example osmoregulation and immune and stress response). Given that migration failure should quickly purge polymorphism at selected SNPs from a population, the question of how genetic diversity in these populations is maintained is an important one. Standing genetic variation could be maintained by different life history strategies and/or environmentally driven balancing selection. Our work highlights the importance of preserving genetic diversity to ensure evolutionary resilience at the population level, and has practical implications for management.

evolutionary biology↗

Genetic consequences of improved river connectivity in brown trout (Salmo trutta, L.)

Fragmentation of watercourses poses a significant threat to biodiversity, particularly for migratory fish species. Mitigation measures such as fishways, have been increasingly implemented to restore river connectivity and support fish migration. The effects of such restoration efforts are typically tested using telemetry and fisheries methods, which do not fully capture the broader population movements that may have important consequences for population viability. We performed a before-and-after control-impact (BACI) study using genetic tools (SNPs) to investigate the effect of a newly implemented fishway, aiming to enhance upstream spawning migration of brown trout (Salmo trutta, Linnaeus) in a reservoir with two headwater tributaries fragmented by man-made weirs. Another reservoir with two barrier-free tributaries was also analysed as a control. Our results showed that the isolated brown trout population was spawning in the reservoir before the installation of the fishway, and we found genetic structuring and differentiation between fragmented headwater tributaries before the fishway construction, but not in the control reservoir. Unexpectedly, after the fishway construction we observed signals consistent with increased genetic differentiation between populations of newly recruited juvenile fish in the reservoir tributary and fish in the reservoir. We propose this was caused by newly enabled philopatric behaviour of brown trout to their natal spawning tributary. In contrast, we did not find any genetic changes in the tributary without a fishway or in the barrier-free reservoir system. Given the scarcity of similar studies, we advocate for an increased use of genetic analyses in BACI studies to monitor and evaluate the effect of efforts to restore habitat connectivity and inform future management strategies.

evolutionary biology↗

Is shape in the eye of the beholder? The reproducibility of geometric morphometric analyses on live animals

Geometric morphometrics is widely used to quantify morphological variation between biological specimens, but the fundamental influence of operator bias on data reproducibility is rarely considered, particularly in studies using photographs of live animals taken under field conditions. We examined this using four independent operators that applied an identical landmarking scheme to replicate photographs of 291 live Atlantic salmon (Salmo salar L.) from two rivers. Using repeated measures tests, we found significant inter-operator differences in mean body shape, suggesting that the operators introduced a systematic error despite following the same landmarking scheme. No significant differences were detected when the landmarking process was repeated by the same operator on a random subset of photographs. Importantly, in spite of significant operator bias, small but statistically significant morphological differences between fish from the two rivers were found consistently by all operators. Pairwise tests of angles of vectors of shape change showed that these between-river differences in body shape were analogous across operator datasets, suggesting a general reproducibility of findings obtained by geometric morphometric studies. In contrast, merging landmark data when fish from each river are digitised by different operators had a significant impact on downstream analyses, highlighting an intrinsic risk of bias. Overall, we show that, even when significant inter-operator error is introduced during digitisation, following an identical landmarking scheme can identify morphological differences between populations. This study indicates that operators digitising at least a sub-set of all data groups of interest may be an effective way of mitigating inter-operator error and potentially enabling data sharing.

zoology↗

Seascape genomics reveals limited dispersal and suggest spatially varying selection among European populations of sea lamprey (Petromyzon marinus)

Sea lamprey Petromyzon marinus is an anadromous and semelparous fish without homing behaviours. Despite being a freshwater, free-living organism for large part of their life cycle, its adulthood is spent as a parasite of marine vertebrates. In their native European range, while it is well-established that sea lampreys comprise a single nearly-panmictic population, few studies have further explored the evolutionary history of natural populations. Here, we performed the first genome-wide characterization of sea lampreys genetic diversity in their European natural range. The objectives were to investigate the connectivity among river basins and explore evolutionary processes mediating dispersal during the marine phase, with the sequencing 186 individuals from 8 locations spanning the North Eastern Atlantic coast and the North Sea with double-digest RAD-sequencing, obtaining a total of 30910 bi-allelic SNPs. Population genetic analyses reinforced the existence of a single metapopulation encompassing freshwater spawning sites within the north eastern Atlantic and the North Sea, though the prevalence of private alleles at northern latitudes suggested some limits to the species dispersal. Seascape genomics suggested a scenario where oxygen concentration and river runoffs impose spatially varying selection across their distribution range. Exploring associations with abundance of potential hosts further suggested that hake and cod could also impose selective pressures, although the nature of such putative biotic interactions was unresolved. Overall, the identification of adaptive seascapes in a panmictic anadromous species could contribute to conservation practices by providing information for restoration activities to mitigate local extinctions on freshwater sites. Sea lamprey Petromyzon marinus is an anadromous and semelparous fish without homing behaviours. Despite being a freshwater, free-living organism for large part of its life cycle, its adulthood is spent as a parasite of marine vertebrates. Here, we performed the first genome-wide characterization of sea lampreys genetic diversity in their European natural range. The objectives were to investigate the connectivity among river basins and explore evolutionary processes mediating dispersal during the marine phase. For that, we sequenced 186 individuals from 8 locations spanning the North-eastern Atlantic coast and the North Sea with double-digest RAD-sequencing, obtaining a total of 30910 bi-allelic SNPs. Population genomic analyses reinforced the existence of a single metapopulation encompassing freshwater spawning sites within the north-eastern Atlantic and the North Sea, though the prevalence of private alleles at northern latitudes suggested some limits to the species dispersal. Seascape analyses revealed candidate loci associated with the abundance of some host species and were located in a genomic region coding for variable lymphocyte receptors, an adaptive immunity tool unique to jawless vertebrates, and to MARCH proteins, a family of E3 ubiquitin ligases also involved in the regulation of immune responses. Abiotic factors (e.g., maximum phosphate, dissolved oxygen and water temperature) were significantly correlated with candidate loci associated with the myo-inositol synthesis, a pathway linked to osmoregulation, and to other genomic regions involved in organismal homeostasis. The identification of adaptive seascapes in this ancient species, especially if linked to primitive adaptive immune responses, could be relevant to understand the evolutionary pathways early in vertebrate evolution.

evolutionary biology↗

Integrating environmental DNA monitoring to inform eel (Anguilla anguilla) status in freshwaters at their easternmost range - A case study in Cyprus

AimDespite significant population declines and targeted EU regulations aimed at A. anguilla conservation, little attention has been given to their status at their easternmost range. This study applies wide scale integrated monitoring to uncover the present-day eel distribution in Cyprus inland freshwaters. These are subject to increasing pressures from water supply requirements and dam construction, as seen throughout the Mediterranean. LocationCyprus. MethodsWe applied environmental DNA metabarcoding of water samples to determine A. anguilla distribution in key freshwater catchments. In addition, we present this alongside ten years of electrofishing/netting data. Refuge traps were also deployed to establish the timing of glass eel recruitment. These outputs are used together, alongside knowledge of the overall fish community and barriers to connectivity, to provide eel conservation and policy insights. ResultsWe confirm the presence of A. anguilla in Cyprus inland freshwaters, with recruitment occurring in March. Eel distribution is restricted to lower elevation areas, and is negatively associated with distance from coast and barriers to connectivity. Many barriers to connectivity are identified, though eels were detected in 2 reservoirs upstream of dams. The overall fish community varies between freshwater habitat types. Main ConclusionsEels are much more widespread in Cyprus than previously thought, yet mostly restricted to lowland intermittent systems. This makes for a case to reconsider the requirement for eel management plans. Environmental DNA based data collected in 2020 indicate that present-day eel distribution is representative of 10-year survey trends. Suggesting that inland freshwaters may act as an unrealised refuge at A. anguillas easternmost range. Conservation efforts in Mediterranean freshwaters should focus on improving connectivity, therefore enabling eels to access inland perennial refugia. Thus, mitigating the impact of climate change and the growing number of fragmented artificially intermittent river systems.

ecology↗