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Too much and not enough data: Challenges and solutions for generating information in freshwater research and monitoring

Evaluating progress toward achieving freshwater conservation and sustainability goals requires transforming diverse types of data into useful information for scientists, managers, and other interest groups. Despite substantial increases in the volume of freshwater data collected worldwide, many regions and ecosystems still lack sufficient data collection and/or data access. We illustrate how these data challenges result from a diverse set of underlying mechanisms and propose solutions that can be applied by individuals or organizations. We discuss creative approaches to address data scarcity, including the use of community science, remote-sensing, environmental sensors, and legacy datasets. We highlight the importance of coordinated data collection efforts among groups and training programs to improve data access. At the institutional level, we emphasize the power of prioritizing data curation, incentivizing data publication, and promoting research that enhances data coverage and representativeness. Some of these strategies involve technological and analytical approaches, but many necessitate shifting the priorities and incentives of organizations such as academic and government research institutions, monitoring groups, journals, and funding agencies. Our overarching goal is to stimulate discussion to narrow the data disparities hindering the understanding of freshwater processes and their change across spatial scales.

Nahuel Huapi Lake, Lake Tahoe

Analysis of the influence of spatial pattern in habitat selection studies

Design and analysis of wildlife habitat selection studies typically do not assess the effect of spatial pattern on the habitat selection process. Effects of landscape scale pattern on habitat selection cannot be accomplished without replicate study areas, because pattern is a single, albeit multifaceted, attribute of an area. For a single area, however, the influence of pattern-related characteristics, such as shape and edge shared with adjacent patches, can be estimated by using GLIM (McCullough and Neider 1983) procedures to model patch-specific frequency counts of animal use as a function of these parameters. This approach is evaluated and illustrated with simulated breeding-bird counts in a South Carolina study area for which a GIS land cover classification is available. A related technique for evaluating whether movement from patch to patch is selective is developed and illustrated for designs that involve collection of trajectory data from monitored individuals. These designs and analyses are feasible given current GIS and GPS technology. Statistical inferences from habitat selection studies should be interpreted within the context of a range of scales at which animals differentiate between patch attributes.

Journal of Agricultural, Biological, and Environme

Seabird tissue archival and monitoring project: Egg collections and analytical results 1999-2002

In 1998, the U.S. Geological Survey Biological Resources Division (USGS-BRD), the U.S. Fish and Wildlife Service (USFWS) Alaska Maritime National Wildlife Refuge (AMNWR), and the National Institute of Standards and Technology (NIST) began the Seabird Tissue Archival and Monitoring Project (STAMP) to collect and cryogenically bank tissues from seabirds in Alaska for future retrospective analysis of anthropogenic contaminants. The approach of STAMP was similar to that of the Alaska Marine Mammal Tissue Archival Project (AMMTAP). AMMTAP was started in 1987 by NIST and the National Oceanic and Atmospheric Administration (NOAA) as part of the Outer Continental Shelf Environmental Assessment Program sponsored by the Minerals Management Service. Presently sponsored by the USGS-BRD, AMMTAP continues its work as part of a larger national program, the Marine Mammal Health and Stranding Response Program. AMMTAP developed carefully designed sampling and specimen banking protocols. Since 1987, AMMTAP has collected tissues from marine mammals taken in Alaska Native subsistence hunts and has cryogenically banked these tissues at the NIST National Biomonitoring Specimen Bank (NBSB). Through its own analytical work and working in partnership with other researchers both within and outside Alaska, AMMTAP has helped to develop a substantial database on contaminants in Alaska marine mammals. In contrast, data and information is limited on contaminants in Alaska seabirds, which are similar to marine mammals in that they feed near the top of the food chain and have the potential for accumulating anthropogenic contaminants. During its early planning stages, STAMP managers identified the seabird egg as the first tissue of choice for study by the project. There is a relatively long history of using bird eggs for environmental monitoring and for investigating the health status of bird populations. Since 1998, protocols for collecting and processing eggs, and cryogenically banking egg samples have been developed by STAMP (see York et al. 2001). Eggs are being collected on an annual basis for several species at nesting colonies throughout Alaska. Aliquots of these egg samples are being analyzed on a regular basis for persistent organic pollutants and mercury. Results of this work have been published in scientific journals (Christopher et al. 2002) and in conference proceedings (Kucklick et al. 2002; Vander Pol et al. 2002a, 2002b). The intent of this report is to provide an up-to-date description of STAMP. The report contains the most recent egg collection inventory, analytical data, preliminary interpretations based on these data, and a discussion of possible future directions of the project.

Report

Environmental conditions associated with bat white-nose syndrome in the north-eastern United States

1. White-nose syndrome (WNS) is an emerging disease of hibernating North American bats that is caused by the cold-growing fungus Geomyces destructans . Since first observed in the winter of 2007, WNS has led to unprecedented mortality in several species of bats and may threaten more than 15 additional hibernating bat species if it continues across the continent. Although the exact means by which fungal infection causes mortality are undetermined, available evidence suggests a strong role of winter environmental conditions in disease mortality. 2. By 2010, the fungus G. destructans was detected in new areas of North America far from the area it was first observed, as well as in eight European bat species in different countries, yet mortality was not observed in many of these new areas of North America or in any part of Europe. This could be because of the differences in the fungus, rates of disease progression and/or in life-history or physiological traits of the affected bat species between different regions. Infection of bats by G. destructans without associated mortality might also suggest that certain environmental conditions might have to co-occur with fungal infection to cause mortality. 3. We tested the environmental conditions hypothesis using Maxent to map and model landscape surface conditions associated with WNS mortality. This approach was unique in that we modelled possible requisite environmental conditions for disease mortality and not simply the presence of the causative agent. 4. The top predictors of WNS mortality were land use/land cover types, mean air temperature of wettest quarter, elevation, frequency of precipitation and annual temperature range. Model results suggest that WNS mortality is most likely to occur in landscapes that are higher in elevation and topographically heterogeneous, drier and colder during winter, and more seasonally variable than surrounding landscapes. 5. Synthesis and applications . This study mapped the most likely environmental surface conditions associated with bat mortality owing to WNS in the north-eastern United Sates; maps can be used for selection of priority monitoring sites. Our results provide a starting point from which to investigate and predict the potential spread and population impacts of this catastrophic emerging disease.

Journal of Applied Ecology

Avian influenza shedding patterns in waterfowl: implications for surveillance, environmental transmission, and disease spread

Despite the recognized importance of fecal/oral transmission of low pathogenic avian influenza (LPAI) via contaminated wetlands, little is known about the length, quantity, or route of AI virus shed by wild waterfowl. We used published laboratory challenge studies to evaluate the length and quantity of low pathogenic (LP) and highly pathogenic (HP) virus shed via oral and cloacal routes by AI-infected ducks and geese, and how these factors might influence AI epidemiology and virus detection. We used survival analysis to estimate the duration of infection (from virus inoculation to the last day virus was shed) and nonlinear models to evaluate temporal patterns in virus shedding. We found higher mean virus titer and longer median infectious period for LPAI-infected ducks (10–11.5 days in oral and cloacal swabs) than HPAI-infected ducks (5 days) and geese (7.5 days). Based on the median bird infectious dose, we found that environmental contamination is two times higher for LPAI- than HPAI-infectious ducks, which implies that susceptible birds may have a higher probability of infection during LPAI than HPAI outbreaks. Less environmental contamination during the course of infection and previously documented shorter environmental persistence for HPAI than LPAI suggest that the environment is a less favorable reservoir for HPAI. The longer infectious period, higher virus titers, and subclinical infections with LPAI viruses favor the spread of these viruses by migratory birds in comparison to HPAI. Given the lack of detection of HPAI viruses through worldwide surveillance, we suggest monitoring for AI should aim at improving our understanding of AI dynamics (in particular, the role of the environment and immunity) using long-term comprehensive live bird, serologic, and environmental sampling at targeted areas. Our findings on LPAI and HPAI shedding patterns over time provide essential information to parameterize environmental transmission and virus spread in predictive epizootiologic models of disease risks.

Journal of Wildlife Diseases

Natural and anthropogenic processes affecting radon releases during mining and early stage reclamation activities, Pinenut uranium mine, Arizona, USA

Radon (Rnair) was monitored in open air in publicly accessible areas surrounding the Pinenut uranium (U) mine during mining and reclamation activities in 2015–16 to address concerns about mining related effects to areas surrounding Grand Canyon National Park (GCNP) in Arizona, USA. During July 2015, Rnair concentrations associated with the ore storage pile monitoring site were larger than those at the mine vent monitoring site and likely resulted from the relatively large amount of ore stored on site during this period. Higher wind velocities at the ore pile monitoring site generally resulted in lower Rnair concentrations; however, wind velocity did not appear to be an important factor in controlling Rnair concentrations at the mine vent monitoring site. Physical disturbances of the ore pile by heavy equipment did not coincide with elevated Rnair concentrations at the ore storage pile or mine vent monitoring sites. The relative size of the ore storage pile showed a positive trend with the daily mean Rnair concentration measured at the ore pile monitoring site. Principal component analysis (PCA) was applied to the ore pile and mine vent multivariate data sets for simultaneous comparison of all measured variables during 230 days of the study period. A significant positive coefficient for Rnair was associated with a significant negative coefficient for wind speed for principal component (PC) 2ore pile. Significant, positive PC2mine vent coefficients included Rnair, wind direction, and relative ore pile size indicating that Rnair variations at the mine vent monitoring site may be affected by Rn sourced from the ore pile. The ore pile is located about 200 m south of the mine vent Rn monitor with the prevalent wind direction coming from the south. All data generated during the field study and laboratory verification tests were published by Naftz et al. (2018) and are available online at:

Journal of Environmental Radioactivity

Global land cover mapping using Earth observation satellite data: Recent progresses and challenges

Land cover is an important variable for many studies involving the Earth surface, such as climate, food security, hydrology, soil erosion, atmospheric quality, conservation biology, and plant functioning. Land cover not only changes with human caused land use changes, but also changes with nature. Therefore, the state of land cover is highly dynamic. In winter snow shields underneath various other land cover types in higher latitudes. Floods may persist for a long period in a year over low land areas in the tropical and subtropical regions. Forest maybe burnt or clear cut in a few days and changes to bare land. Within several months, the coverage of crops may vary from bare land to nearly 100% crops and then back to bare land following harvest. The highly dynamic nature of land cover creates a challenge in mapping and monitoring which remains to be adequately addressed. As economic globalization continues to intensify, there is an increasing trend of land cover/land use change, environmental pollution, land degradation, biodiversity loss at the global scale, timely and reliable information on global land cover and its changes is urgently needed to mitigate the negative impact of global environment change.

ISPRS Journal of Photogrammetry and Remote Sensing

Simplified method for detecting tritium contamination in plants and soil

Cost-effective methods are needed to identify the presence and distribution of tritium near radioactive waste disposal and other contaminated sites. The objectives of this study were to (i) develop a simplified sample preparation method for determining tritium contamination in plants and (ii) determine if plant data could be used as an indicator of soil contamination. The method entailed collection and solar distillation of plant water from foliage, followed by filtration and adsorption of scintillation-interfering constituents on a graphite-based solid phase extraction (SPE) column. The method was evaluated using samples of creosote bush [ Larrea tridentata (Sessé & Moc. ex DC.) Coville], an evergreen shrub, near a radioactive disposal area in the Mojave Desert. Laboratory tests showed that a 2-g SPE column was necessary and sufficient for accurate determination of known tritium concentrations in plant water. Comparisons of tritium concentrations in plant water determined with the solar distillation–SPE method and the standard (and more laborious) toluene-extraction method showed no significant difference between methods. Tritium concentrations in plant water and in water vapor of root-zone soil also showed no significant difference between methods. Thus, the solar distillation–SPE method provides a simple and cost-effective way to identify plant and soil contamination. The method is of sufficient accuracy to facilitate collection of plume-scale data and optimize placement of more sophisticated (and costly) monitoring equipment at contaminated sites. Although work to date has focused on one desert plant, the approach may be transferable to other species and environments after site-specific experiments.

Journal of Environmental Quality

Evaluating the waterfowl breeding population and habitat survey for scaup

Potential bias in breeding population estimates of certain duck species from the Waterfowl Breeding Population and Habitat Survey (WBPHS) has been a concern for decades. The WBPHS does not differentiate between lesser ( Aythya affinis ) and greater ( A . marila ) scaup, but lesser scaup comprise 89% of the combined scaup population and their population estimates are suspected to be biased. We marked female lesser scaup (i.e., marked scaup) in the Mississippi and Atlantic Flyways, Canada and United States, with implantable satellite transmitters to track their spring migration through the traditional and eastern survey areas of the WBPHS, 2005–2010. Our goal was to use data independent of the WBPHS to evaluate whether breeding population estimates for scaup were biased and identify variables that might be used in the future to refine population estimates. We found that the WBPHS estimates of breeding scaup are biased because, across years, only 30% of our marked scaup had settled for the breeding period when the strata in which they settled were surveyed, 43% were available to be counted in multiple survey strata as their migration continued during the WBPHS, 32% settled outside the WBPHS area, the number of times a marked scaup was available to be counted by survey crews varied positively with the latitude that a marked scaup settled on breeding areas, the probability of a marked scaup being in a stratum while it was surveyed varied among years, and these probabilities were positively correlated with the traditional and eastern breeding population estimates for scaup. Annual population estimates derived from banding data provide a less biased and preferable method of monitoring scaup population status and trend. Development of models that include metrics such as survey stratum latitude and annual spring environmental conditions might potentially be used to improve scaup breeding population estimates derived from the WBPHS, but independent estimates from banding data would be important to evaluate such models.

Journal of Wildlife Management

Blood lead exposure concentrations in mottled ducks ( Anas fulvigula ) on the upper Texas coast

The mottled duck (Anas fulvigula) is a non-migratory waterfowl species dependent upon coastal marsh systems, including those on the Texas Chenier Plain National Wildlife Refuge (NWR) Complex, and considered a regional indicator species of marsh habitat quality. Research from the early 1970s, 1990s, and early-2000s indicated that mottled ducks continued to exhibit elevated wing-bone lead (Pb) concentrations, decades after implementation of non-toxic shot regulations. However, wing-bone concentrations reflect lifetime accumulation of Pb, whereas blood Pb concentrations reflect more recent exposure. To identify current potentially relevant temporal windows of Pb exposure, we collected 260 blood samples from mottled ducks during summer (n=124) and winter (n=136) from 2010–2012 on the Texas Chenier Plain NWR Complex. We quantified baseline blood Pb concentrations for all ages of mottled ducks, and hypothesized that blood lead concentrations would remain elevated above background levels (200 µg L–1) despite the 1983 and 1991 lead shot bans. Blood Pb concentrations ranged from below detection limits to >12,000 µg L–1, where >200 µg L–1 was associated with exposure levels above background concentrations. Male mottled ducks had the greatest blood Pb concentrations (30 times greater than females) with concentrations greater during winter than summer. Likewise, the proportion of exposed (>200 µg L–1) females increased from 14%–47% from summer to winter, respectively. Regardless of sex, adult mottled duck blood Pb concentrations were five times greater than juveniles, particularly during winter. We identified five plausible models that influenced blood Pb levels where year, site, and interactions among age*sex*season and between age*season were included in the top-ranked models. Frequency of exposure was greatest during winter, increasing from 12% in summer to 55% in winter, indicating that a temporal exposure window to environmental Pb exists between nesting and hunting seasons. Blood Pb concentrations remain elevated in mottled ducks despite Pb shot bans enacted >25 years prior to this study. If Pb levels in mottled ducks becomes a conservation concern, regional monitoring of blood Pb concentrations would be appropriate with a focus upon elucidating potential reasons for the variation among age and sex groups. Finally, identifying potentially available sources of environmental Pb may be key to minimizing this apparently persistent threat to mottled ducks on the upper Texas coast.

Journal of the Southeastern Association of Fish an

Preliminary evaluation of the floating dome method of measuring reaeration rates

The floating dome method of measuring reaeration rates was evaluated using a modified tracer technique and a disturbed equilibrium technique. The lack of agreement between results of the floating dome method and the other methods was attributed to limitations in instrument accuracy amplified by physical relationships of the dome to the water column rather than conceptual errors in the dome method. In techniques where polarographic oxygen probes are used to monitor the oxygen content within the dome atmosphere, the ratio of volume of the dome to volume of the water column under the dome must be kept below 0.02 to minimize instrument error to less than 10 percent of the predicted oxygen transport. The combination of instrument limitations, coupled with environmental limitations caused by wind and the greenhouse effects, severely restrict application of the floating dome method.

Journal of Research of the U.S. Geological Survey

Compensatory recruitment unlikely in high-elevation amphibian populations challenged with disease

Understanding the causes of population variation in host response to disease, and the mechanisms of persistence, can serve as vital information for species conservation. One such mechanism of population persistence that has gained support is the demographic process of compensatory recruitment. Host populations may persist by increasing recruitment to compensate for reduced survival due to infection, thus limiting the negative effects of the disease on population trajectories. However, high-elevation populations are inherently vulnerable to stochastic processes and may be limited in their ability to exhibit compensatory recruitment relative to lower elevation populations. We use long-term mark–recapture data from five populations of boreal toads Anaxyrus boreas boreas , across an elevational gradient in Colorado, before and after pathogen arrival to assess whether populations can persist with Batrachochytrium dendrobatidis ( Bd ) via compensatory recruitment. Prior to pathogen arrival, we found a life-history trade-off between survival and recruitment across elevations, where high-elevation toads have high survival but lower recruitment and vice versa at lower elevations. Pathogen arrival had a strong negative effect on apparent annual survival and recruitment leading to negative population growth rates and dramatically reduced host abundances. The data did not support the occurrence of compensatory recruitment. Synthesis and applications . Our unique dataset indicates that demographic responses to pathogens may be environmentally (i.e. elevationally) context dependent and highlights the value of long-term monitoring. We recommend that practitioners verify that potential persistence mechanisms occur across multiple populations and relevant environmental gradients to counter any assumptions of the mechanism existing species-wide. Quantifying variation in population responses to disease will aid in understanding the bounds of such persistence mechanisms and identify particularly vulnerable populations where mechanisms are nonexistent.

Journal of Applied Ecology

Coastal fine-grained sediment plumes from beach nourishment near Santa Barbara, California

Terrestrial sediments captured by flood control facilities such as dams, debris basins, and engineered stream channels can reduce sediment fluxes to littoral cells. The beneficial use of these sediments for beach nourishment may induce negative environmental effects from turbidity or sedimentation caused by the source material. Here, we examine the size and extent of turbid coastal plumes produced by beach nourishment with sediment containing significant fine-grained (silt and clay) fractions near Santa Barbara, California, to evaluate the potential effects of beneficial use of fluvial sediment captured in upstream facilities. Using remotely sensed imagery and hydrodynamic and sediment transport model simulations, we find that wave height, wind speed and direction, and sediment settling velocity have strong controls on the direction and extent of the turbid plume produced by beach nourishment. These results are consistent with monitored nourishment projects, suggesting generalizable patterns and processes across the studies. Additionally, we find that sediment placement method (hauling versus dredging) can influence the magnitude and duration of fine-sediment concentrations in the adjacent coastal waters. Combined, these results will inform sediment management strategies that intend to minimize the environmental effects of beach nourishment within the study area and across similar coastal sites.

California

Interaction of ambient conditions and fecal coliform bacteria in southern Lake Michigan beach waters: Monitoring program implications

Excessive fecal coliform bacteria in public swimming waters can potentially threaten visitor health. Fecal coliform bacteria (1984-1989) and Escherichia coli (1990-1995) density were monitored weekly at the Indiana Dunes National Lakeshore beaches for 12 summers, and park officials closed swimming areas when fecal coliform density exceeded the state water quality criteria (400 CFU fecal coliforms/ 100 ml; 235 CFU E. coli/100 ml water). Due to a 24-hour incubation in the fecal coliform and E. coli assays, beaches were closed the day after collection of high fecal coliform. Our analysis suggests that it is not possible to predict one day's fecal coliform count based on the previous day's results in waters taken from southern Lake Michigan beaches. Dispersal and deposition of bacteria were not uniform among sites or across time apparently due to interactions among environmental variables including rainfall, wind direction, water temperature, and bacteria source. Rainfall combined with northwest winds increased bacteria concentrations. Escherichia coli followed a seasonal trend with similar fluctuations in density among beaches. We suggest that the current beach monitoring protocol is inadequate for predicting fecal coliform density at the time of beach closure, and, subsequently, its use for ensuring visitor safety remains questionable.

Indiana

Status of the major aquaculture carps of China in the Laurentian Great Lakes Basin

There is concern of economic and environmental damage occuring if any of the four major aquacultured carp species of China, black carp Mylopharyngodon piceus , bighead carp Hypophthalmichthys nobilis , silver carp H. molitrix , or grass carp Ctenopharyngodon idella , were to establish in the Laurentian Great Lakes. All four are reproducing in the Mississippi River Basin. We review the status of these fishes in relation to the Great Lakes and their proximity to pathways into the Great Lakes, based on captures and collections of eggs and larvae. No black carp have been captured in the Great Lakes Basin. One silver carp and one bighead carp were captured within the Chicago Area Waterway System, on the Great Lakes side of electric barriers designed to keep carp from entering the Great Lakes from the greater Mississippi River Basin. Three bighead carp were captured in Lake Erie, none later than the year 2000. By December 2019, at least 650 grass carps had been captured in the Great Lakes Basin, most in western Lake Erie, but none in Lake Superior. Grass carp reproduction has been documented in the Sandusky and Maumee rivers in Ohio, tributaries of Lake Erie. We also discuss environmental DNA (eDNA) results as an early detection and monitoring tool for bighead and silver carps. Detection of eDNA does not necessarily indicate presence of live fish, but bigheaded carp eDNA has been detected on the Great Lakes side of the barriers and in a small proportion of samples from the western basin of Lake Erie.

Illinois, Indiana

Groundwater and surface-water exchange and resultingnNitrate dynamics in the Bogue Phalia Basin in northwestern Mississippi

During April 2007 through September 2008, the USGS collected hydrogeologic and water-quality data from a site on the Bogue Phalia to evaluate the role of groundwater and surface-water interaction on the transport of nitrate to the shallow sand and gravel aquifer underlying the Mississippi Alluvial Plain in northwestern Mississippi. A two-dimensional groundwater/surface-water exchange model was developed using temperature and head data and VS2DH, a variably saturated flow and energy transport model. Results from this model showed that groundwater/surface-water exchange at the site occurred regularly and recharge was laterally extensive into the alluvial aquifer. Nitrate was consistently reported in surface-water samples ( n = 52, median concentration = 39.8 μmol/L) although never detected in samples collected from in-stream piezometers or shallow monitoring wells adjacent to the stream ( n = 46). These two facts, consistent detections of nitrate in surface water and no detections of nitrate in groundwater, coupled with model results that indicate large amounts of surface water moving through an anoxic streambed, support the case for denitrification and nitrate loss through the streambed.

Mississippi

Residues of organochlorines and heavy metals in ruddy ducks from the Delaware River, 1973

In December 1973, eight ruddy ducks killed in an oil spill on the Delaware River were collected to be analyzed for residues of environmental pollutants. Whole carcasses were analyzed for organochlorine pesticides and livers were examined for lead, cadmium, and mercury. Residues of polychlorinated biphenyls and DDT and/or its metabolites were present in all carcasses. Dieldrin and hexachlorobenzene were present in seven of the eight samples. All livers contained detectable levels of lead, cadmium, and mercury.

Pesticides Monitoring Journal

Metabolic rates in an anadromous clupeid, the American shad ( Alosa sapidissima )

To assess the energetics of migration in an anadromous fish, adult American shad ( Alosa sapidissima ) were swum in a large respirometer at a range of speeds (1.0–2.3 body lengths (BL) s −1 , 13–24 °C). Metabolic rate (M O2 ) was logarithmically related to swimming speed (Bl s −1 ; r 2 = 0.41, slope = 0.23 ± 0.037) and tailbeat frequency (beats × min −1 ; r 2 = 0.52, slope = 0.003 ± 0.0003). Temperature had a significant effect on metabolic rate ( r 2 = 0.41) with a Q 10 of 2.2. Standard metabolic rate (SMR), determined directly after immobilization with the neuroblocker gallamine triethiodide, ranged from 2.2–6.2 mmolO 2 kg −1 h −1 and scaled with mass (W) such that SMR = 4.0 (±0.03)W 0.695(±0.15) . Comparison of directly determined and extrapolated SMR suggests that swimming respirometry provides a good estimate of SMR in this species, given the differences in basal activity monitored by the two methods. Overall, American shad metabolic rates (M O2 and SMR) were intermediate between salmonids and fast-swimming perciforms, including tunas, and may be a result of evolutionary adaptation to their active pelagic, schooling life history. This study demonstrates variability in metabolic strategy among anadromous fishes that may be important to understanding the relative success of different migratory species under varying environmental conditions.

Journal of Comparative Physiology B: Biochemical,