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Evaluation of rapid DNA extraction methods to better enable point-of-use environmental DNA detection

Recent developments in molecular testing have created the opportunity for biologists and managers to detect environmental DNA (eDNA) of target species rapidly and without the requirement of a laboratory. These point-of-use protocols may be especially useful for early detection and rapid response for invasive species or surveillance for at-risk native species, where timely management decisions are critical. Point-of-use eDNA protocols also facilitate wider and less expensive implementation of eDNA methods. One of the key components to an effective point-of-use protocol is a rapid DNA extraction method. Several rapid extraction protocols are suitable for implementation in the field, but information regarding their relative effectiveness is lacking. We evaluated extraction efficiency of four DNA rapid extraction protocols using filters spiked with primary cultured grass carp ( Ctenopharyngodon idella ) gill cells. The extraction methods included two syringe-based column extractions, a lysis and extraction solution, and a divalent cation chelation resin (Chelex) extraction protocol alongside a laboratory-based control kit. We estimated DNA yield using a newly designed quantitative polymerase chain reaction (qPCR) assay targeting the grass carp nuclear genome. We evaluated two additional factors, filter type (mixed cellulose ester [MCE] and polyethersulfone [PES]) and background eDNA source (aquaculture or river). The lysis and extraction solution and Chelex extraction both had the highest overall yield, with MCE filters further increasing Chelex yield while the enzyme extraction yield was dependent on interaction with both filter and eDNA source. Our results indicate that rapid extraction protocols, such as solutions with short heating steps, are effective for DNA isolation and help to increase the overall accessibility of eDNA analyses.

Environmental DNA

The oilspill risk analysis model of the U. S. Geological Survey

The U.S. Geological Survey has developed an oilspill risk analysis model to aid in estimating the environmental hazards of developing oil resources in Outer Continental Shelf (OCS) lease areas. The large, computerized model analyzes the probability of spill occurrence, as well as the likely paths or trajectories of spills in relation to the locations of recreational and biological resources which may be vulnerable. The analytical methodology can easily incorporate estimates of weathering rates , slick dispersion, and possible mitigating effects of cleanup. The probability of spill occurrence is estimated from information on the anticipated level of oil production and method and route of transport. Spill movement is modeled in Monte Carlo fashion with a sample of 500 spills per season, each transported by monthly surface current vectors and wind velocities sampled from 3-hour wind transition matrices. Transition matrices are based on historic wind records grouped in 41 wind velocity classes, and are constructed seasonally for up to six wind stations. Locations and monthly vulnerabilities of up to 31 categories of environmental resources are digitized within an 800,000 square kilometer study area. Model output includes tables of conditional impact probabilities (that is, the probability of hitting a target, given that a spill has occurred), as well as probability distributions for oilspills occurring and contacting environmental resources within preselected vulnerability time horizons. (USGS)

Open-File Report

The oilspill risk analysis model of the U. S. Geological Survey

The U.S. Geological Survey has developed an oilspill risk analysis model to aid in estimating the environmental hazards of developing oil resources in Outer Continental Shelf (OCS) lease areas. The large, computerized model analyzes the probability of spill occurrence, as well as the likely paths or trajectories of spills in relation to the locations of recreational and biological resources which may be vulnerable. The analytical methodology can easily incorporate estimates of weathering rates , slick dispersion, and possible mitigating effects of cleanup. The probability of spill occurrence is estimated from information on the anticipated level of oil production and method of route of transport. Spill movement is modeled in Monte Carlo fashion with a sample of 500 spills per season, each transported by monthly surface current vectors and wind velocities sampled from 3-hour wind transition matrices. Transition matrices are based on historic wind records grouped in 41 wind velocity classes, and are constructed seasonally for up to six wind stations. Locations and monthly vulnerabilities of up to 31 categories of environmental resources are digitized within an 800,000 square kilometer study area. Model output includes tables of conditional impact probabilities (that is, the probability of hitting a target, given that a spill has occurred), as well as probability distributions for oilspills occurring and contacting environmental resources within preselected vulnerability time horizons. (USGS)

Professional Paper

Hydrogeologic framework of fractured sedimentary rock, Newark Basin, New Jersey

The hydrogeologic framework of fractured sedimentary bedrock at the former Naval Air Warfare Center (NAWC), Trenton, New Jersey, a trichloroethylene (TCE)-contaminated site in the Newark Basin, is developed using an understanding of the geologic history of the strata, gamma-ray logs, and rock cores. NAWC is the newest field research site established as part of the U.S. Geological Survey Toxic Substances Hydrology Program, Department of Defense (DoD) Strategic Environmental Research and Development Program, and DoD Environmental Security Technology Certification Program to investigate contaminant remediation in fractured rock. Sedimentary bedrock at the NAWC research site comprises the Skunk Hollow, Byram, and Ewing Creek Members of the Lockatong Formation and Raven Rock Member of the Stockton Formation. Muds of the Lockatong Formation that were deposited in Van Houten cycles during the Triassic have lithified to form the bedrock that is typical of much of the Newark Basin. Four lithotypes formed from the sediments include black, carbon-rich laminated mudstone, dark-gray laminated mudstone, light-gray massive mudstone, and red massive mudstone. Diagenesis, tectonic compression, off-loading, and weathering have altered the rocks to give some strata greater hydraulic conductivity than other strata. Each stratum in the Lockatong Formation is 0.3 to 8 m thick, strikes N65 degrees E, and dips 25 degrees to 70 degrees NW. The black, carbon-rich laminated mudstone tends to fracture easily, has a relatively high hydraulic conductivity and is associated with high natural gamma-ray count rates. The dark-gray laminated mudstone is less fractured and has a lower hydraulic conductivity than the black carbon-rich laminated mudstone. The light-gray and the red massive mudstones are highly indurated and tend to have the least fractures and a low hydraulic conductivity. The differences in gamma-ray count rates for different mudstones allow gamma-ray logs to be used to correlate and delineate the lithostratigraphy from multiple wells. Gamma-ray logs and rock cores were correlated to develop a 13-layer gamma-ray stratigraphy and 41-layer lithostratigraphy throughout the fractured sedimentary rock research site. Detailed hydrogeologic framework shows that black carbon-rich laminated mudstones are the most hydraulically conductive. Water-quality and aquifer-test data indicate that groundwater flow is greatest and TCE contamination is highest in the black, carbon- and clay-rich laminated mudstones. Large-scale groundwater flow at the NAWC research site can be modeled as highly anisotropic with the highest component of permeability occurring along bedding planes.

New Jersey

Evolution of Ore Deposits and Technology Transfer Project: Isotope and Chemical Methods in Support of the U.S. Geological Survey Science Strategy, 2003-2008

Principal functions of the U.S. Geological Survey (USGS) Mineral Resources Program are providing assessments of the location, quantity, and quality of undiscovered mineral deposits, and predicting the environmental impacts of exploration and mine development. The mineral and environmental assessments of domestic deposits are used by planners and decisionmakers to improve the stewardship of public lands and public resources. Assessments of undiscovered mineral deposits on a global scale reveal the potential availability of minerals to the United States and other countries that manufacture goods imported to the United States. These resources are of fundamental relevance to national and international economic and security policy in our globalized world economy. Performing mineral and environmental assessments requires that predictions be made of the likelihood of undiscovered deposits. The predictions are based on geologic and geoenvironmental models that are constructed for the diverse types of mineral deposits from detailed descriptions of actual deposits and detailed understanding of the processes that formed them. Over the past three decades the understanding of ore-forming processes has benefited greatly from the integration of laboratory-based geochemical tools with field observations and other data sources. Under the aegis of the Evolution of Ore Deposits and Technology Transfer Project (referred to hereinafter as the Project), a 5-year effort that terminated in 2008, the Mineral Resources Program provided state-of-the-art analytical capabilities to support applications of several related geochemical tools to ore-deposit-related studies. The analytical capabilities and scientific approaches developed within the Project have wide applicability within Earth-system science. For this reason the Project Laboratories represent a valuable catalyst for interdisciplinary collaborations of the type that should be formed in the coming years for the United States to meet its natural-resources and natural-science needs. This circular presents an overview of the Project. Descriptions of the Project laboratories are given first including descriptions of the types of chemical or isotopic analyses that are made and the utility of the measurements. This is followed by summaries of select measurements that were carried out by the Project scientists. The studies are grouped by science direction. Virtually all of them were collaborations with USGS colleagues or with scientists from other governmental agencies, academia, or the private sector.

Circular

Paleoecological insights on fixed tree island development in the Florida Everglades: I. environmental controls

Palynological and geochemical analyses of sediment cores collected on two tree islands in the Florida Everglades indicate long-term hydrologic and chemical differences between tree islands and surrounding marshes and sloughs. Gumbo Limbo and Nuthouse tree islands are elongate, teardrop-shaped islands in Water Conservation Area 3B. Prior to tree island formation at both sites, pollen records indicate that sites on modern tree island heads were covered with sawgrass marshes with abundant weedy annuals. Such vegetation is characteristic of moderate water depths and hydroperiods with frequent droughts or disturbances. Contemporaneously deposited sediments on tree island tails indicate progressively deeper water conditions with increasing distance from the head; wetlands surrounding tree islands were covered by sloughs with deep water and long hydroperiods. Tree island formation occurred at about 1200 BC on Gumbo Limbo Island, with mature tree island vegetation established by about 800 AD. On Nuthouse Island, tree island formation occurred around 300 AD, shifting to mature tree island vegetation around 1400 AD. Thus, tree island formation began on these islands between 3.2 Ka and 1.7 Ka. Maturation of tree islands took between 1,000 and 2,000 years, and vegetation on these tree islands has been relatively stable for the last 600–1,200 years. Phosphorus levels on tree island heads have been extremely high (approximately six times greater than baseline levels in marshes) throughout the history of the sites, and phosphorus content in tree island tails began increasing when tree island formation occurred. Elevated phosphorus content may reflect the long-term presence of wading birds at these sites and provide a proxy for reconstructing the historic distribution of wading bird populations.

Florida

Resource and land information for south Dade County, Florida

South Dade County, at the southeast corner of Florida, exemplifies many of the environmental concerns that have developed throughout the United States. Considerable environmental information has recently been assembled from this region, and some of it is presented in this experimental report in folio form.

Florida

Ground-water models as a management tool in Florida

Highly sophisticated computer models provide powerful tools for analyzing historic data and for simulating future water levels, water movement, and water chemistry under stressed conditions throughout the ground-water system in Florida. Models that simulate the movement of heat and subsidence of land in response to aquifer pumping also have potential for application to hydrologic problems in the State. Florida, with 20 ground-water modeling studies reported since 1972, has applied computer modeling techniques to a variety of water-resources problems. Models in Florida generally have been used to provide insight to problems of water supply, contamination, and impact on the environment. The model applications range from site-specific studies, such as estimating contamination by wastewater injection at St. Petersburg, to a regional model of the entire State that may be used to assess broad-scale environmental impact of water-resources development. Recently, groundwater models have been used as management tools by the State regulatory authority to permit or deny development of water resources. As modeling precision, knowledge, and confidence increase, the use of ground-water models will shift more and more toward regulation of development and enforcement of environmental laws. (USGS)

Water-Resources Investigations Report

Environmental Characteristics and Geographic Information System Applications for the Development of Nutrient Thresholds in Oklahoma Streams

The U.S.Environmental Protection Agency has developed nutrient criteria using ecoregions to manage and protect rivers and streams in the United States. Individual states and tribes are encouraged by the U.S. Environmental Protection Agency to modify or improve upon the ecoregion approach. The Oklahoma Water Resources Board uses a dichotomous process that stratifies streams using environmental characteristics such as stream order and stream slope. This process is called the Use Support Assessment Protocols, subchapter15. The Use Support Assessment Protocols can be used to identify streams threatened by excessive amounts of nutrients, dependant upon a beneficial use designation for each stream. The Use Support Assessment Protocols, subchapter 15 uses nutrient and environmental characteristic thresholds developed from a study conducted in the Netherlands, but the Oklahoma Water Resources Board wants to modify the thresholds to reflect hydrologic and ecological conditions relevant to Oklahoma streams and rivers. Environmental characteristics thought to affect impairment from nutrient concentrations in Oklahoma streams and rivers were determined for 798 water-quality sites in Oklahoma. Nutrient, chlorophyll, water-properties, and location data were retrieved from the U.S. Environmental Protection Agency STORET database including data from the U.S. Geological Survey, Oklahoma Conservation Commission, and Oklahoma Water Resources Board. Drainage-basin area, stream order, stream slope, and land-use proportions were determined for each site using a Geographic Information System. The methods, procedures, and data sets used to determine the environmental characteristics are described.

Water-Resources Investigations Report

Physical condition and stress levels during early development reflect feeding rates and predict pre- and post-fledging survival in a nearshore seabird

The effects of acute environmental stressors on reproduction in wildlife are often difficult to measure because of the labour and disturbance involved in collecting accurate reproductive data. Stress hormones represent a promising option for assessing the effects of environmental perturbations on altricial young; however, it is necessary first to establish how stress levels are affected by environmental conditions during development and whether elevated stress results in reduced survival and recruitment rates. In birds, the stress hormone corticosterone is deposited in feathers during the entire period of feather growth, making it an integrated measure of background stress levels during development. We tested the utility of feather corticosterone levels in 3- to 4-week-old nestling brown pelicans ( Pelecanus occidentalis ) for predicting survival rates at both the individual and colony levels. We also assessed the relationship of feather corticosterone to nestling body condition and rates of energy delivery to nestlings. Chicks with higher body condition and lower corticosterone levels were more likely to fledge and to be resighted after fledging, whereas those with lower body condition and higher corticosterone levels were less likely to fledge or be resighted after fledging. Feather corticosterone was also associated with intracolony differences in survival between ground and elevated nest sites. Colony-wide, mean feather corticosterone predicted nest productivity, chick survival and post-fledging dispersal more effectively than did body condition, although these relationships were strongest before fledglings dispersed away from the colony. Both reproductive success and nestling corticosterone were strongly related to nutritional conditions, particularly meal delivery rates. We conclude that feather corticosterone is a powerful predictor of reproductive success and could provide a useful metric for rapidly assessing the effects of changes in environmental conditions, provided pre-existing baseline variation is monitored and understood.

Gulf of Mexico

Development of a quantitative PCR assay for detecting Egeria densa in environmental DNA samples

Brazilian elodea ( Egeria densa ) is an invasive freshwater plant that demonstrates widespread ecological impacts in freshwater ecosystems and causes substantial economic damage. Here, we developed an environmental DNA assay for detection of E. densa to provide resource managers with a tool for early detection, identification, and monitoring of invasive populations.

Conservation Genetics Resources

A python framework for environmental model uncertainty analysis

We have developed pyEMU, a python framework for Environmental Modeling Uncertainty analyses, open-source tool that is non-intrusive, easy-to-use, computationally efficient, and scalable to highly-parameterized inverse problems. The framework implements several types of linear (first-order, second-moment (FOSM)) and non-linear uncertainty analyses. The FOSM-based analyses can also be completed prior to parameter estimation to help inform important modeling decisions, such as parameterization and objective function formulation. Complete workflows for several types of FOSM-based and non-linear analyses are documented in example notebooks implemented using Jupyter that are available in the online pyEMU repository. Example workflows include basic parameter and forecast analyses, data worth analyses, and error-variance analyses, as well as usage of parameter ensemble generation and management capabilities. These workflows document the necessary steps and provides insights into the results, with the goal of educating users not only in how to apply pyEMU, but also in the underlying theory of applied uncertainty quantification.

Environmental Modelling and Software

Environmental geochemistry and sediment quality in Lake Pontchartrain: database development and review

This paper reports on preliminary results of a project to develop a comprehensive data base of chemical and environmental information on sediments from Lake Pontchartrain, Louisiana, and surrounding water bodies. The goal is to evaluate all data for reliability and comparability, and to make it widely accessible and useful to all users. Methods for processing heterogeneous, historical data follow previous methods employed in the Boston Harbor and Massachusetts Bay area. Data from 11 different data sets, encompassing about 900 total samples, have been entered to date. Questionable or anomalous data were noted in a minority of cases. Problems tend to follow distinct patterns and are relatively easy to identify. Hence, comparability of data has not proven to be the major obstacle to synthesis efforts that was anticipated in earlier years (NRC, 1989). Quality-controlled data sets show that the bulk of sediment samples in the more central parts of Lake Pontchartrain have values within normal background for heavy metals like Cu, Pb, and Zn. The same or lower concentrations were found in the vicinity of the Bonnet Carre Spillway, representing influx from the Mississippi River. Mean concentrations for Cu, Pb, and Zn were 17, 21, and 74 µg/g (total dissolution analyses), respectively. However, values as high as 267 µg/g Pb and comparable increases for other metal and organic contaminants are found in sediments within 2 km of the coastal strip of New Orleans. Additional sampling in such areas and in other inland coastal waterways is needed, since such levels are above the threshold for potential toxic effects on benthic organisms, according to effects-based screening criteria. The most contaminated sites, Bayou Trepagnier and Bayou Bonfouca, involve industrial areas where waste discharge has now been controlled or remediated, but where sediments may retain large concentrations of contaminants, e.g. tenths of a percent of Pb, Cr, and Zn or more for Bayou Trepagnier.

Louisiana

A socio-environmental geodatabase for integrative research in the transboundary Rio Grande/Río Bravo basin

Management of water resources in the transboundary Rio Grande/Río Bravo Basin (the Basin) presents challenges for state and Federal entities in the United States and Mexico making management decisions on shared water resources. Damming, channelization, water availability, and allocation are governed by water rights and water-sharing agreements. Data and information sharing are important aspects of transboundary cooperation, but differences in format, content, spatial and temporal resolution, and language hinder collaboration. In addition, data on the kinds and geographic distribution of water governance and management institutions across the Basin have not been consistently documented. Existing data disparities parallel the hydrological and social fragmentation of the Basin. Seeking to underscore the interdependence between social and environmental processes in the Basin, anthropologists and modelers collaborated to develop a socio-environmental geodatabase. This geodatabase is a first step in modeling the social components of decision making and their connectivity to environmental processes across the Basin. The geodatabase is available in an open-access domain and contains geospatial data related to water and land governance, hydrology, water use and hydraulic infrastructure, socioeconomics, and the biophysical environment necessary to advance the understanding of basin dynamics. Having these data documented and compiled in a central location serves as a resource to help decision makers better understand upstream and downstream social-environmental characteristics. This knowledge is useful for developing sustainable water management policies in a region where water resources are increasingly under pressure from climatic, environmental, and human-related changes.

Chihuahua, New Mexico, Texas

Biogeographic freshwater fish pattern legacy revealed despite rapid socio-economic changes in China

Understanding drivers of freshwater fish assemblages is critically important for biodiversity conservation strategies, especially in rapidly developing countries, which often have environmental protections lagging behind economic development. The influences of natural and human factors in structuring fish assemblages and their relative contributions are likely to change given the increasing magnitude of human activities. To discriminate natural and human drivers of fish diversity and assemblage patterns in developing countries with rapid socio-economic development, a dataset of 908 freshwater fish species and 13 metrics including three categories of both natural (i.e., biogeographic) and human drivers (i.e., economic growth, inland fisheries) in China were analysed with machine learning algorithms (i.e., self-organizing map, random forest). Here, we found that biogeographic drivers explained 21.8% of the observed fish assemblage patterns in China and remained stronger predictors when compared to human drivers (i.e., 15.6%, respectively). Freshwater fish species richness was positively correlated to rainfall, air temperature, surface water area and inland fisheries production but negatively correlated with urbanization. In addition, the strong structuring effects of climatic variables on Chinese fish richness patterns suggested that the fish assemblages could be particularly vulnerable to climate change. Our results showed that natural biogeographic factors still dominate in driving freshwater fish assemblage patterns despite increased human disturbances on aquatic ecosystems in a rapidly developing country. These findings consequently suggested that we should consider both natural (e.g., climate) and human (e.g., urbanization, inland fisheries) factors when establishing aquatic conservation strategies and priorities for developing countries that are experiencing rapid socio-economic changes.

Fish and Fisheries

Quantifying water requirements of riparian river red gum ( Eucalyptus camaldulensis ) in the Murray-Darling Basin, Australia: Implications for the management of environmental flows

Water resource development and drought have altered river flow regimes, increasing average flood return intervals across floodplains in the Murray-Darling Basin, Australia, causing health declines in riparian river red gum ( Eucalyptus camaldulensis ) forests and woodlands. Environmental flow allocations helped to alleviate water stress during the recent Millennium Drought (1997–2010), however, quantification of the flood frequency required to support healthy E. camaldulensis communities is still needed. We quantified water requirements of E. camaldulensis for two years across a flood gradient (trees inundated at frequencies of 1:2, 1:5 and 1:10 years) at Yanga National Park, New South Wales to help inform management decision-making and design of environmental flows. Sap flow, evaporative losses and soil moisture measurements were used to determine transpiration, evapotranspiration and plant-available soil water before and after flooding. A formula was developed using plant-available soil water post-flooding and average annual rainfall, to estimate maintenance time of soil water reserves in each flood frequency zone. Results indicated that soil water reserves could sustain 1:2 and 1:5 trees for 15 months and six years, respectively. Trees regulated their transpiration rates, allowing them to persist within their flood frequency zone, and showed reduction in active sapwood area and transpiration rates when flood frequencies exceeded 1:2 years. A leaf area index of 0.5 was identified as a potential threshold indicator of severe drought stress. Our results suggest environmental water managers may have greater flexibility to adaptively manage floodplains in order to sustain E. camaldulensis forests and woodlands than has been appreciated hitherto.

New South Wales

Landscape conservation design for enhancing the adaptive capacity of coastal wetlands in the face of sea-level rise and coastal development

Coastal wetlands provide many valuable benefits to people and wildlife, including critical habitat, improved water quality, reduced flooding impacts, and protected coastlines. However, in the 21st century, accelerated sea-level rise and coastal development are expected to greatly alter coastal landscapes across the globe. The future of coastal wetlands is uncertain, challenging coastal environmental managers to develop conservation strategies that will increase the resilience of these valuable ecosystems to change and preserve the benefits they provide. One strategy for preparing for the effects of sea-level rise is to ensure that there is space available for coastal wetlands to migrate inland. In a recent study, we identified areas where coastal wetlands may move inland along the northern Gulf of Mexico coast, one of the most wetland-rich and sea-level rise sensitive regions of the world. Building on these findings, this project produced customized landscape conservation-design products focused on identifying landward migration routes for coastal wetlands. The resulting products provide environmental managers with information to make decisions to enhance the capacity of coastal wetlands to adapt to sea-level rise and coastal development, protecting these ecosystems and the critical economic and ecological benefits that they provide.

Alabama, Florida, Louisiana, Mississippi, Texas

Critical environmental areas program workshops: A synopsis of results

Problems besetting the development of State Critical Environmental Areas Programs were discussed at a series of six workshops sponsored by the Resource and Land Investigations (RALI) Program and Office of Land Use and Water Policy (OLUWP) of the Department of the Interior. The major problems were determined to be: program justification, effective utilization of existing institutions, the individuality of State needs, the length of time over which benefits would accrue to such programs, the roles and interactions of different levels of government, an acceptable measure for program success, public involvement in program development, information and data handling problems, and program priorities. The role of the Federal Government was thought to be a critical factor in developing such programs, especially in the western States where Federal land ownership may subordinate State planning efforts. Priorities in Critical Environmental Areas Programs were seen to be closely tied to concern for preservation of ways-of-life, a concern which broadens the Critical Environmental Areas concept from mainly environmental to societal in scope. The utility of the workshop format as a means of idea exchange is discussed in light of these examples.

Open-File Report