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At least 829 records · Page 46Linked to original sources
USGS2014

Next-generation genomic shotgun sequencing indicates greater genetic variability in the mitochondria of Hypophthalmichthys molitrix relative to H. nobilis from the Mississippi River, USA and provides tools for research and detection

We characterized variation within the mitochondrial genomes of the invasive silver carp ( Hypophthalmichthys molitrix ) and bighead carp ( H. nobilis ) from the Mississippi River drainage by mapping our Next-Generation sequences to their publicly available genomes. Variant detection resulted in 338 single-nucleotide polymorphisms for H. molitrix and 39 for H. nobilis . The much greater genetic variation in H. molitrix mitochondria relative to H. nobilis may be indicative of a greater North American female effective population size of the former. When variation was quantified by gene, many tRNA loci appear to have little or no variability based on our results whereas protein-coding regions were more frequently polymorphic. These results provide biologists with additional regions of DNA to be used as markers to study the invasion dynamics of these species.

Conservation Genetics Resources↗
USGS2023

Validation of a species-specific probe-based qPCR assay for the threatened meltwater stonefly, Lednia tumana, in environmental samples

A probe-based quantitative real-time PCR assay was developed to detect meltwater stonefly ( Lednia tumana ) environmental (e)DNA in water samples. The limits of detection and quantification, respectively, were 12.1 and 58.4 gene copies for calibration standards and these values were similarly low in a relevant environmental sample matrix (8.6 and 174.2, respectively). The assay’s utility was demonstrated in situ on water samples with concomitant manual invertebrate surveys from a wide range of alpine streams across L. tumana ’s native range.

Conservation Genetics Resources↗
USGS2023

Development of a quantitative PCR assay for detecting northwest salamander (ambystoma gracile) in environmental DNA samples

We developed a primer and probe based quantitative PCR assay for use with environmental DNA to detect Northwest salamander ( Ambystoma gracile ), a species endemic to the temperate Pacific coastal region of North America. The assay targets a region in the mitochondrial DNA (mtDNA) D-loop gene. Tests of the assay were performed in silico (using the NCBI BLAST tool), in vitro (using DNA extracted from A. gracile and related species) and in situ (water samples collected from a lake with a known population of A. gracile ). This assay will be useful in efforts to monitor the distribution and occupancy of this species.

Conservation Genetics Resources↗
USGS1991

Heterosis and outbreeding depression: A multi-locus model and an application to salmon production

Both artificial propagation and efforts to preserve or augment natural populations sometimes involve, wittingly or unwittingly, the mixing of different gene pools. The advantages of such mixing vis-à-vis the alleviation of inbreeding depression are well known. Acknowledged, but less well understood, are the complications posed by outbreeding depression. This paper derives a simple model of outbreeding depression and demonstrates that it is reasonably possible to predict the generation-to-generation fitness course of hybrids derived from parents from different origins. Genetic difference, or distance between parental types, is defined by the drop in fitness experienced by one type reared at the site to which the other is locally adapted. For situations where decisions involving stock mixing must be made in the absence of complete information, a sensitivity analysis-based conflict resolution method (the Good-Bad-Ugly model) is described.

Fisheries Research↗
USGS2017

Genetic responses to rapid change in the environment during the anthropocene

Humans have greatly affected the genetic composition of many different organisms during the Anthropocene. Humans cause genetic changes by affecting the direction and magnitude of evolutionary forces that act to create the Earth's biota. In many cases, we expect the outcome of human actions to be extinction and hybridization of existing species, but other outcomes, such as adaptation, also occur. Given the reach of humans throughout the globe, and recent biotechnology advances that make it possible to move individual genes between species or to remove them, it is likely that human influence on the genetic composition of other organisms will become even more widespread as the Anthropocene progresses.

Book chapter↗
USGS2015

Evaluating the status of individuals and populations: Advantages of multiple approaches and time scales

The assessment of population status is a central goal of applied wildlife research and essential to the field of wildlife conservation. “Population status” has a number of definitions, the most widely used having to do with the current trajectory of the population (i.e., growing, stable, or declining), or the probability of persistence (i.e., extinction risk), perhaps without any specific knowledge as to the factors driving a population’s dynamics. In contrast, a population’s status relative to the carrying capacity of the environment ( K ) is an ecologically-based definition that explicitly provides information about a major mechanism of population control. That is, it relates to the relative per capita availability of resources to individuals in a population, which can also be used to infer the state of the environment itself. Sea otters in the North Pacific provide an excellent system with which to examine various approaches to assessing population status relative to K . This is because sea otters were nearly extirpated by historic commercial overexploitation in the eighteenth and nineteenth centuries, followed by natural and translocation-aided population recovery during the twentieth century, and this decline and recovery has been relatively well documented. This provided a unique opportunity to study populations at the extremes of the population status spectrum. Here we describe and review the approaches that have been utilized in sea otter research to understand the status of populations relative to resource abundance. Specifically, we will illustrate the utility of various indices of population status for understanding population dynamics using the case study of a second precipitous sea otter decline in the Western Aleutians. The indices or “tools” described here fit into several broad categories including (1) energetic, (2) morphological, and (3) demographic as well as a fourth category of emerging tools that have not yet been employed in many other situations including dietary diversity, community structure, spatial distribution, and gene expression. Overall, a variety of indices used to measure population status throughout the sea otter’s range have provided insights for understanding the mechanisms driving the trajectory of various sea otter populations, which a single index could not, and we suggest using multiple methods to measure a population’s status at multiple spatial and temporal scales. The work described here also illustrates the usefulness of long-term data sets and/or approaches that can be used to assess population status retrospectively, providing information otherwise not available. While not all systems will be as amenable to using all the approaches presented here, we expect innovative researchers could adapt analogous multi-scale methods to a broad range of habitats and species including apex predators occupying the top trophic levels, which are often of conservation concern.

Book chapter↗
USGS2016

Paramyxoviruses of fish

The first fish paramyxovirus was isolated from normal adult Chinook salmon returning to a coastal hatchery in Oregon in the fall of 1982. Subsequently, the virus was isolated from other stocks of adult Chinook salmon and one stock of adult coho salmon in California, Oregon, Washington and Alaska, leading to its designation as the Pacific salmon paramyxovirus ( PSPV ). The slow-growing virus can be isolated from tissues and ovarian fluids of healthy adult fish returning to spawn and apparently causes no clinical signs of disease or mortality. In 1995, a different and widely disseminated paramyxovirus was isolated from farmed Atlantic salmon in Norway and was designated as Atlantic salmon paramyxovirus ( ASPV ). Although this virus caused no disease or mortality when injected into juvenile Atlantic salmon, ASPV has been associated with proliferative gill inflammation in sea-reared yearling fish; however, additional infectious agents may be involved in the etiology of the condition. Sequence analysis of PSPV and ASPV isolates using the polymerase gene established their placement in the family Paramyxoviridae and has shown the two viruses to be closely related but sufficiently different from each other and from other known paramyxoviruses to possibly represent new genera within the family. The viruses can be diagnosed by isolation in cell culture with final confirmation by molecular methods. Other paramyxovirus-like agents have been observed or isolated from rainbow trout in Germany, from seabream in Japan associated with epithelial necrosis, from turbot in Spain associated with erythrocytic inclusion bodies and buccal/opercular hemorrhaging and from koi and common carp associated with gill necrosis in the European Union.

Book chapter↗
USGS2024

Paramyxoviruses of fish

The first fish paramyxovirus was isolated from normal adult Chinook salmon returning to a coastal hatchery in Oregon in the fall of 1982. Subsequently, the virus was isolated from other stocks of adult Chinook salmon and one stock of adult coho salmon in California, Oregon, Washington, and Alaska, leading to its designation as the Pacific salmon paramyxovirus (PsaPV). The slow-growing virus can be isolated from tissues and ovarian fluids of healthy adult fish returning to spawn and apparently causes no clinical signs of disease or mortality. In 1995 a different and widely disseminated paramyxovirus was isolated from farmed Atlantic salmon in Norway and was designated as Atlantic salmon paramyxovirus (AsaPV). Although this virus caused no disease or mortality when injected into juvenile Atlantic salmon, AsaPV has been associated with proliferative gill inflammation in sea-reared yearling fish; however, additional infectious agents may be involved in the etiology of the condition. Sequence analysis of PsaPV and AsaPV isolates using the polymerase gene established their placement in the family Paramyxoviridae and has shown the two viruses to be closely related but sufficiently different from each other and from other known paramyxoviruses to represent a new genus within the family. The viruses can be diagnosed by isolation in cell culture with final confirmation by molecular methods. Other paramyxovirus-like agents have been observed or isolated from rainbow trout in Germany, from seabream in Japan associated with epithelial necrosis, from turbot in Spain associated with erythrocytic inclusion bodies and buccal/opercular hemorrhaging and from koi and common carp associated with gill necrosis in the European Union.

Book chapter↗
USGS2026

Using quantitative polymerase chain reaction to assess phytoplankton and indicate eutrophication in freshwater rivers: A multiyear nationwide study across the United States

Phytoplankton are essential primary producers in fresh surface water that are critical to the health of ecosystems. However, phytoplankton overgrowth due to eutrophication threatens ecological, economic, and public health. Therefore, assessing phytoplankton is fundamental for understanding the productivity, health, and trophic status of freshwater ecosystems. Light microscopy and chlorophyll a assessment are common approaches for studying phytoplankton. They are easy to use, cost-effective, and reliable but have significant limitations. Microscopy has a low throughput and is time-consuming and labor-intensive. Chlorophyll a assessment does not reveal phytoplankton community composition and structure. For comparison, quantitative polymerase chain reaction (qPCR) is widely applied in quantifying microorganisms, offering multiple advantages, including high throughput, sensitivity, accuracy, and robustness. However, a research gap remains regarding the feasibility of using qPCR to assess phytoplankton and indicate trophic status of freshwater bodies. We conducted a nationwide, multiyear study in the United States to compare the performance of qPCR, microscopy, and chlorophyll a assessment in assessing phytoplankton and trophic statuses of multiple freshwater rivers. From early summer to late fall in 2017, 2018, and 2019, we assessed phytoplankton, chlorophyll a , pheophytin a , and the overall Trophic Level Index ( TLI Overall ) at the sampling sites in 12 large freshwater rivers in three regions (western, midcontinent, and eastern) across the United States. The seasonal summed abundance of four major phytoplankton taxa [Bacillariophyta (diatoms), Cyanobacteria (blue-green algae), Chlorophyta (green algae), and Dinoflagellates (Dinophyta)] ranged from 6.88 log 10 (GCN·L –1 ) (the Connecticut River, 2017) to 9.29 log 10 (GCN·L –1 ) (the Kansas River, 2019) (GCN: gene or genome copy number). qPCR- and microscopy-based phytoplankton abundance of eight phytoplankton taxa had a significant positive allometric or log-linear correlation (adjusted R 2 = 0.836, p -value < 0.001, n = 815). In addition, qPCR-based phytoplankton abundance had positive allometric or log-linear correlations with chlorophyll a (adjusted R 2 = 0.5437, p -value < 0.001, n = 164), pheophytin a (adjusted R 2 = 0.3378, p -value < 0.001, n = 164), and TLI Overall (adjusted R 2 = 0.4789, p -value < 0.001, n = 164). Therefore, qPCR is a promising alternative to microscopy and chlorophyll a for studying phytoplankton and trophic status in freshwater rivers. Moreover, phytoplankton abundance had limited temporal variation within each sampling season and over the three sampling seasons in 2017, 2018, and 2019 but showed clear spatial variation. The midcontinent sites had significantly higher phytoplankton abundance, chlorophyll a concentrations, pheophytin a concentrations, and TLI Overall values than those in the eastern and western rivers, reflecting the higher trophic statuses of the midcontinent rivers. This work also provides the thresholds of qPCR-based phytoplankton abundance for delineating trophic statuses in freshwater rivers. Overall, this work demonstrates that qPCR is a promising tool for studying phytoplankton and characterizing the trophic status of freshwater rivers.

Book chapter↗
USGS2016

Cultivation of a native alga for biomass and biofuel accumulation in coal bed methane production water

Coal bed methane (CBM) production has resulted in thousands of ponds in the Powder River Basin of low-quality water in a water-challenged region. A green alga isolate, PW95, was isolated from a CBM production pond, and analysis of a partial ribosomal gene sequence indicated the isolate belongs to the Chlorococcaceae family. Different combinations of macro- and micronutrients were evaluated for PW95 growth in CBM water compared to a defined medium. A small level of growth was observed in unamended CBM water (0.15 g/l), and biomass increased (2-fold) in amended CBM water or defined growth medium. The highest growth rate was observed in CBM water amended with both N and P, and the unamended CBM water displayed the lowest growth rate. The highest lipid content (27%) was observed in CBM water with nitrate, and a significant level of lipid accumulation was not observed in the defined growth medium. Growth analysis indicated that nitrate deprivation coincided with lipid accumulation in CBM production water, and lipid accumulation did not increase with additional phosphorus limitation. The presented results show that CBM production wastewater can be minimally amended and used for the cultivation of a native, lipid-accumulating alga.

Montana, Wyoming↗
USGS2013

Assessing the state of knowledge of utility-scale wind energy development and operation on non-volant terrestrial and marine wildlife

A great deal has been published in the scientific literature regarding the effects of wind energy development and operation on volant (flying) wildlife including birds and bats, although knowledge of how to mitigate negative impacts is still imperfect. We reviewed the peer-reviewed scientific literature for information on the known and potential effects of utility-scale wind energy development and operation (USWEDO) on terrestrial and marine non-volant wildlife and found that very little has been published on the topic. Following a similar review for solar energy we identified known and potential effects due to construction and eventual decommissioning of wind energy facilities. Many of the effects are similar and include direct mortality, environmental impacts of destruction and modification of habitat including impacts of roads, and offsite impacts related to construction material acquisition, processing and transportation. Known and potential effects due to operation and maintenance of facilities include habitat fragmentation and barriers to gene flow, as well as effects due to noise, vibration and shadow flicker, electromagnetic field generation, macro- and micro-climate change, predator attraction, and increased fire risk. The scarcity of before-after-control-impact studies hinders the ability to rigorously quantify the effects of USWEDO on non-volant wildlife. We conclude that more empirical data are currently needed to fully assess the impact of USWEDO on non-volant wildlife.

Applied Energy↗
USGS2021

Exposure to Deepwater Horizon crude oil increases free cholesterol in larval red drum (Sciaenops ocellatus)

The 2010 Deepwater Horizon oil spill impacted over 2100 km of shoreline along the northern Gulf of Mexico, which coincided with the spawning season of many coastal species, including red drum ( Sciaenops ocellatus ). Red drum develop rapidly and are sensitive to crude oil exposure during the embryonic and larval periods. This study investigates the predictions from recent transcriptomic studies that cholesterol biosynthetic processes are impacted by oil exposure in fish early life stages. We found that red drum larvae exposed for 72-hours to ΣPAH 50 3.55 - 15.45 µg L −1 exhibited significantly increased pericardial area, a cardiotoxicity metric, but the expression of several genes targeted in the cholesterol synthesis pathway was not affected. However, whole-mount staining revealed significant increases in free cholesterol throughout the larval body (ΣPAH 50 4.71 – 16.15 µg L −1 ), and total cholesterol followed an increasing trend (ΣPAH 50 3.55 - 15.45 µg L −1 ). Cholesterol plays a critical role in fish embryo development and ion channel function. Therefore, the disruption of cholesterol homeostasis, as observed here, could play a role in the oil toxicity phenotype observed across many fish species.

Aquatic Toxicology↗
USGS2025

Establishment of a cell culture from Daphnia magna as an in vitro model for (eco)toxicology assays: Case study using Bisphenol A as a representative cytotoxic and endocrine disrupting chemical

Bisphenol A (BPA) is a widely used industrial compound found in polycarbonate plastics, epoxy resin, and various polymer materials, leading to its ubiquitous presence in the environment. The toxicity of BPA to aquatic organisms has been well documented following in vivo exposure scenarios, with known cytotoxic and endocrine-disrupting effects. As such, BPA was used in this study as a well-characterized chemical to implement more ethical and resource-efficient scientific practices in toxicity testing through new approach methods (NAMs). Due to the frequent use of Daphnia spp. as a model organism in toxicology, we developed an in vitro cell culture system from Daphnia magna embryos, with optimized medium to support cell longevity. The cultures were maintained for up to two months, demonstrating their stability and suitability for cytotoxicity studies. Using this novel system, lethal concentration 50 (LC 50 ) values were determined at the 24 and 48 h time points following BPA exposure. Subsequently, oxidative stress, endocrine disruption, and DNA damage were assessed through gene expression, activity assays, and a comet assay in BPA-exposed cells. LC 50 values of 52 µM and 20 µM BPA were calculated after 24 and 48 h exposures, respectively. BPA cells exposed to 20 and 52 µM had significantly increased GSH, GPx, and GST activity levels. mRNA expression analysis revealed significant upregulations in the expression of hsp70, hsp90, gst, gpx, vtg1 , and cyp4 , with downregulations of sod, cat , and ecr following BPA exposure. Furthermore, comet assays showed a significantly higher level of DNA damage induced by BPA compared to controls, with greater comet and tail lengths. This study established a novel in vitro Daphnia model, using BPA as a case study for determining toxic effects, further highlighting the importance and applicability of utilizing alternative methods in ecotoxicological research through reducing animal use.

Aquatic Toxicology↗
USGS2025

Occurrence and tissue distribution of per- and polyfluoroalkyl substances (PFAS) in fishes from waterbodies with point and non-point sources in Massachusetts, USA

Per- and polyfluoroalkyl substances (PFAS) are persistent environmental contaminants with known bioaccumulative and toxic effects in aquatic ecosystems. This study assessed site-specific differences in PFAS contamination in fish from Ashumet Pond, Sudbury River, and Great Herring Pond (reference site) in Massachusetts. Fish from Ashumet Pond exhibited the highest PFAS concentrations, particularly perfluorooctane sulfonate (PFOS), which exceeded levels in plasma almost 650 times those at the reference site. Principal component analysis identified distinct PFAS profiles at each site, reflecting localized contamination sources. Temporal analysis at Ashumet Pond revealed a substantial increase in plasma PFOS and perfluorodecanoic acid (PFDA) from 2020 to 2022. Tissue distribution analyses showed the highest PFAS concentrations in plasma, followed by liver and muscle, consistent with PFAS binding affinity for blood proteins. Species-specific differences in PFAS bioaccumulation were observed, with largemouth bass ( Micropterus nigricans ) exhibiting higher body burdens than banded killifish ( Diaphanus fundulus ), likely due to trophic position and dietary exposure. Histopathological assessments and gene transcript analyses revealed associations between PFAS exposure and inflammatory responses, oxidative stress, endocrine disruption, and immune-related pathways, with the most pronounced molecular effects observed at the downstream site of the Sudbury River. This study underscores the importance of understanding site-specific contamination sources, exposure pathways, and biological effects of PFAS in fish. These findings would benefit from additional research on sediment contamination, temporal analyses at each site, trophic transfer, and transcriptomic analyses across multiple organs to further elucidate PFAS toxicity mechanisms and guide remediation efforts.

Massachusetts↗
USGS2016

Acid rain and its environmental effects: Recent scientific advances

The term ‘acid rain’ refers to atmospheric deposition of acidic constituents that impact the earth as rain, snow, particulates, gases, and vapor. Acid rain was first recognized by Ducros (1845) and subsequently described by the English chemist Robert Angus Smith (Smith, 1852) whose pioneering studies linked the sources to industrial emissions and included early observations of deleterious environmental effects (Smith, 1872). Smith's work was largely forgotten until the mid-20th century when observations began to link air pollution to the deposition of atmospheric sulfate (SO 4 2− ) and other chemical constituents, first near the metal smelter at Sudbury, Ontario, Canada, and later at locations in Europe, North America, and Australia (Gorham, 1961). Our modern understanding of acid rain as an environmental problem caused largely by regional emissions of sulfur dioxide (SO 2 ) and nitrogen oxides (NO x ) stems from observations in the 1960s and early 1970s in Sweden by Svante Odén (Odén, 1976), and in North America by Gene Likens and colleagues (Likens and Bormann, 1974). These scientists and many who followed showed the link to emissions from coal-fired power plants and other industrial sources, and documented the environmental effects of acid rain such as the acidification of surface waters and toxic effects on vegetation, fish, and other biota.

Atmospheric Environment↗
USGS2011

Conservation genetics of evolutionary lineages of the endangered mountain yellow-legged frog, Rana muscosa (Amphibia: Ranidae), in southern California

Severe population declines led to the listing of southern California Rana muscosa (Ranidae) as endangered in 2002. Nine small populations inhabit watersheds in three isolated mountain ranges, the San Gabriel, San Bernardino and San Jacinto. One population from the Dark Canyon tributary in the San Jacinto Mountains has been used to establish a captive breeding population at the San Diego Zoo Institute for Conservation Research. Because these populations may still be declining, it is critical to gather information on how genetic variation is structured in these populations and what historical inter-population connectivity existed between populations. Additionally, it is not clear whether these populations are rapidly losing genetic diversity due to population bottlenecks. Using mitochondrial and microsatellite data, we examine patterns of genetic variation in southern California and one of the last remaining populations of R. muscosa in the southern Sierra Nevada. We find low levels of genetic variation within each population and evidence of genetic bottlenecks. Additionally, substantial population structure is evident, suggesting a high degree of historical isolation within and between mountain ranges. Based on estimates from a multi-population isolation with migration analysis, these populations diversified during glacial episodes of the Pleistocene, with little gene flow during population divergence. Our data demonstrate that unique evolutionary lineages of R. muscosa occupy each mountain range in southern California and should be managed separately. The captive breeding program at Dark Canyon is promising, although mitigating the loss of neutral genetic diversity relative to the natural population might require additional breeding frogs.

California↗
USGS2013

Localized extinction of an arboreal desert lizard caused by habitat fragmentation

We adopted a species’ perspective for predicting extinction risk in a small, endemic, and strictly scansorial lizard ( Urosaurus nigricaudus ), in an old (∼60 year) and highly fragmented (8% habitat remaining) agricultural landscape from the Sonoran Desert, Mexico. We genotyped 10 microsatellite loci in 280 individuals from 11 populations in fragmented and continuous habitat. Individual dispersal was restricted to less than 400 m, according to analyses of spatial autocorrelation and spatially explicit Bayesian assignment methods. Within this scale, continuous areas and narrow washes with native vegetation allowed high levels of gene flow over tens of kilometers. In the absence of the native vegetation, cleared areas and highways were identified as partial barriers. In contrast, outside the scale of dispersal, cleared areas behaved as complete barriers, and surveys corroborated the species went extinct after a few decades in all small (less than 45 ha), isolated habitat fragments. No evidence for significant loss of genetic diversity was found, but results suggested fragmentation increased the spatial scale of movements, relatedness, genetic structure, and potentially affected sex-biased dispersal. A plausible threshold of individual dispersal predicted only 23% of all fragments in the landscape were linked with migration from continuous habitat, while complete barriers isolated the majority of fragments. Our study suggested limited dispersal, coupled with an inability to use a homogeneous and hostile matrix without vegetation and shade, could result in frequent time-delayed extinctions of small ectotherms in highly fragmented desert landscapes, particularly considering an increase in the risk of overheating and a decrease in dispersal potential induced by global warming.

Sonoran Desert↗
USGS2014

Population declines lead to replicate patterns of internal range structure at the tips of the distribution of the California red-legged frog ( Rana draytonii )

Demographic declines and increased isolation of peripheral populations of the threatened California red-legged frog ( Rana draytonii ) have led to the formation of internal range boundaries at opposite ends of the species’ distribution. While the population genetics of the southern internal boundary has been studied in some detail, similar information is lacking for the northern part of the range. In this study, we used microsatellite and mtDNA data to examine the genetic structuring and diversity of some of the last remaining R. draytonii populations in the northern Sierra Nevada, which collectively form the northern external range boundary. We compared these data to coastal populations in the San Francisco Bay Area, where the species is notably more abundant and still exists throughout much of its historic range. We show that ‘external’ Sierra Nevada populations have lower genetic diversity and are more differentiated from one another than their ‘internal’ Bay Area counterparts. This same pattern was mirrored across the distribution in California, where Sierra Nevada and Bay Area populations had lower allelic variability compared to those previously studied in coastal southern California. This genetic signature of northward range expansion was mirrored in the phylogeography of mtDNA haplotypes; northern Sierra Nevada haplotypes showed greater similarity to haplotypes from the south Coast Ranges than to the more geographically proximate populations in the Bay Area. These data cast new light on the geographic origins of Sierra Nevada R. draytonii populations and highlight the importance of distinguishing the genetic effects of contemporary demographic declines from underlying signatures of historic range expansion when addressing the most immediate threats to population persistence. Because there is no evidence of contemporary gene flow between any of the Sierra Nevada R. draytonii populations, we suggest that management activities should focus on maintaining and creating additional ponds to support breeding within typical dispersal distances of occupied habitat.

Biological Conservation↗