Geology ReportsSearch

Geology topics

Gaia Meigs-Friend

Publications and source records attributed to Gaia Meigs-Friend.

8 recordsLinked to original sources

Environmental DNA surveys of Burmese pythons in the Greater Everglades Ecosystem

Improving the probability of detecting invasive giant snakes is vital for the management of emerging or established populations. Burmese pythons occupy thousands of square kilometers of mostly inaccessible habitats in Florida. Environmental DNA (eDNA) methods have been shown to be time and cost effective in a number of systems and may be preferable to traditional detection methods for constrictor snakes, having been shown to be effective at detecting Burmese pythons where traditional and novel detection methods have failed. The purposes of this study were (1) to estimate Burmese python eDNA occurrence in the Greater Everglades Ecosystem based on land-use type; and (2) to conduct preliminary surveys within the Greater Everglades Ecosystem for positive eDNA detections. Twenty-eight sites were sampled in the Greater Everglades Ecosystem, with 5 field replicate samples per site, for a total of 140 water samples collected. Python eDNA was detected in samples from 25 of the 28 sites by using droplet digital polymerase chain reaction amplification. Abiotic parameters were collected and explored, but we found no conclusive relationship among them and python eDNA detections. eDNA monitoring of aquatic habitats can assist in identifying newly colonized areas where pythons have not been previously detected, as well as movement corridors and pathways of dispersal. This information could be used to delimit a population boundary as it expands further to the north in peninsular Florida.

Florida

Improving eDNA yield and inhibitor reduction through increased water volumes and multi-filter isolation techniques

To inform management and conservation decisions, environmental DNA (eDNA) methods are used to detect genetic material shed into the water by imperiled and invasive species. Methodological enhancements are needed to reduce filter clogging, PCR inhibition, and false-negative detections when eDNA is at low concentrations. In the first of three simple experiments, we sought to ameliorate filter clogging from particulates and organic material through a scaled-up, multi-filter protocol. We combined four filters in a 5 mL Phenol-Chloroform-Isoamyl (PCI) procedure to allow for larger volumes of water (~1 L) to be filtered rapidly. Increasing the filtered water volume by four times resulted in 4.4X the yield of target DNA. Next, inhibition from organic material can reduce or block eDNA detections in PCR-based assays. To remove inhibitory compounds retained during eDNA isolation, we tested three methods to chemically strip inhibitors from eDNA molecules. The use of CTAB as a short-term (5–8 day) storage buffer, followed by a PCI isolation, resulted in the highest eDNA yields. Finally, as opposed to a linear relationship among increasing concentrations of filtered genomic eDNA, we observed a sharp change between the lower (70–280 ng) and higher (420–560 ng) amounts. This may be important for effectively precipitating eDNA during protocol testing.

Scientific Reports

Efficacy of eDNA as an early detection indicator for Burmese pythons in the ARM Loxahatchee National Wildlife Refuge in the Greater Everglades Ecosystem

Environmental DNA (eDNA) detection of invasive species can be used to delimited occupied ranges and estimate probabilities to inform management decisions. Environmental DNA is shed into the environment through skin cells and bodily fluids and can be detected in water samples collected from lakes, rivers, and swamps. In south Florida, invasive Burmese pythons occupy much of the Greater Everglades in mostly inaccessible habitat and are credited with causing severe declines of native species’ populations. Detection of Burmese pythons by traditional methods, such as trapping and visual searching, have been largely ineffective, making eDNA a superior method for differentiating invaded habitat. We adapted a quantitative PCR eDNA assay for droplet digital PCR, a state-of-the-art method that improves precision and accuracy. From August 2014 to October 2016, locations in and around Arthur R. Marshall Loxahatchee National Wildlife Refuge in southeast Florida were surveyed for Burmese python eDNA. The Refuge is maintained to provide water storage and is considered one of the last remnants of the northern Everglades wetlands. Positive eDNA detections were made at each of the five sampling events, assessing a total of 399 samples, with moderate occurrence (ψ=58-91%) and detection (p=40-70%) probabilities, potentially reduced by high PCR inhibition-levels. The high occurrence rates and geographic distribution of the positive samples within the Refuge suggests a steady release of python eDNA from a resident Burmese python population and reduces support for primarily transport of eDNA through boats or flowing water from the north. The first confirmed sighting of a Burmese python in the Refuge occurred in September 2016, after eDNA testing had indicated the presence of pythons. An established population is not expected this far north, however, the detections likely indicate northern range limit of a consistent population at Loxahatchee on the eastern side of the Florida peninsula. Our study demonstrates the benefit of eDNA for determining more accurate range limits and expansion information for Burmese pythons, as well as laying the foundation for the assessment of control efforts.

Florida

Environmental DNA (eDNA) detection of nonnative bullseye snakehead in southern Florida

Bullseye Snakehead Channa marulius (Hamilton 1822) was first detected in the southern Florida town of Tamarac in 2000 and has been expanding its geographic range since. Environmental DNA (eDNA) analysis is a newly-developed technique used to noninvasively detect cryptic or low-density species or those that are logistically difficult-to-study. Genetic material shed into the environment through tissue and body fluids is concentrated from water samples and analyzed for the presence of target species eDNA. To help delineate Bullseye Snakehead’s geographic range, we developed and validated a species-specific eDNA assay for both quantitative and droplet digital PCR (ddPCR). We then used ddPCR to assess 16 locations in southeast Florida using 222 water samples collected from 2015 to 2018. Positive eDNA detections were obtained at all six locations that were within the known geographic range of Bullseye Snakehead. Furthermore, eDNA was detected in six of 10 locations that were previously thought to be outside the periphery of the range but hydrologically connected through the extensive canal system. Over the four years of sampling, estimated occurrence rates (ψ) remained stable and relatively high (ψ = 0.67 [95% credible interval (CI) 0.33–0.95]) near Tamarac, Florida, as compared to the most southern sampling locations (ψ = 0.0–0.37). Bulls- eye Snakehead eDNA estimated occurrence rates in the middle region increased between 2016 (0.28 [95% CI 0.03–0.94]) and 2017 (0.66 [95% CI 0.24–0.98]), potentially reflecting eDNA detections related to a growing or expanding population. Bullseye Snakehead eDNA was detected at low concentrations on the northern and eastern borders of Everglades National Park, which is an important conservation area and UNESCO World Heritage Site. Despite extensive sampling via electrofishing, no Bullseye Snakehead were visually detected in several locations that yielded positive eDNA samples. It is unclear whether eDNA was transported through flowing water or another vector. To date, collection records for this species are confined to urban canals; however, Bullseye Snakehead may use the interconnected system of canals to disperse to natural conservation areas such as Everglades National Park, Big Cypress National Preserve, and Water Conservation Areas, where it may impact native species via predation and competition.

Florida

Surveys of environmental DNA (eDNA): a new approach to estimate occurrence in Vulnerable manatee populations

Environmental DNA (eDNA) detection is a technique used to non-invasively detect cryptic, low density, or logistically difficult-to-study species, such as imperiled manatees. For eDNA measurement, genetic material shed into the environment is concentrated from water samples and analyzed for the presence of target species. Cytochrome b quantitative PCR and droplet digital PCR eDNA assays were developed for the 3 Vulnerable manatee species: African, Amazonian, and both subspecies of the West Indian (Florida and Antillean) manatee. Environmental DNA assays can help to delineate manatee habitat ranges, high use areas, and seasonal population changes. To validate the assay, water was analyzed from Florida’s east coast containing a high-density manatee population and produced 31564 DNA molecules l -1 on average and high occurrence (ψ) and detection (p) estimates (ψ = 0.84 [0.40-0.99]; p = 0.99 [0.95-1.00]; limit of detection 3 copies µl -1 ). Similar occupancy estimates were produced in the Florida Panhandle (ψ = 0.79 [0.54-0.97]) and Cuba (ψ = 0.89 [0.54-1.00]), while occupancy estimates in Cameroon were lower (ψ = 0.49 [0.09-0.95]). The eDNA-derived detection estimates were higher than those generated using aerial survey data on the west coast of Florida and may be effective for population monitoring. Subsequent eDNA studies could be particularly useful in locations where manatees are (1) difficult to identify visually (e.g. the Amazon River and Africa), (2) are present in patchy distributions or are on the verge of extinction (e.g. Jamaica, Haiti), and (3) where repatriation efforts are proposed (e.g. Brazil, Guadeloupe). Extension of these eDNA techniques could be applied to other imperiled marine mammal populations such as African and Asian dugongs.

Endangered Species Research

Model description and evaluation of the mark-recapture survival model used to parameterize the 2012 status and threats analysis for the Florida manatee ( Trichechus manatus latirostris )

This report provides supporting details and evidence for the rationale, validity and efficacy of a new mark-recapture model, the Barker Robust Design, to estimate regional manatee survival rates used to parameterize several components of the 2012 version of the Manatee Core Biological Model (CBM) and Threats Analysis (TA). The CBM and TA provide scientific analyses on population viability of the Florida manatee subspecies ( Trichechus manatus latirostris ) for U.S. Fish and Wildlife Service’s 5-year reviews of the status of the species as listed under the Endangered Species Act. The model evaluation is presented in a standardized reporting framework, modified from the TRACE (TRAnsparent and Comprehensive model Evaluation) protocol first introduced for environmental threat analyses. We identify this new protocol as TRACE-MANATEE SURVIVAL and this model evaluation specifically as TRACE-MANATEE SURVIVAL, Barker RD version 1. The longer-term objectives of the manatee standard reporting format are to (1) communicate to resource managers consistent evaluation information over sequential modeling efforts; (2) build understanding and expertise on the structure and function of the models; (3) document changes in model structures and applications in response to evolving management objectives, new biological and ecological knowledge, and new statistical advances; and (4) provide greater transparency for management and research review.

Open-File Report

Detection limits of quantitative and digital PCR assays and their influence in presence-absence surveys of environmental DNA

A set of universal guidelines is needed to determine the limit of detection (LOD) in PCR-based analyses of low concentration DNA. In particular, environmental DNA (eDNA) studies require sensitive and reliable methods to detect rare and cryptic species through shed genetic material in environmental samples. Current strategies for assessing detection limits of eDNA are either too stringent or subjective, possibly resulting in biased estimates of species’ presence. Here, a conservative LOD analysis grounded in analytical chemistry is proposed to correct for overestimated DNA concentrations predominantly caused by the concentration plateau, a nonlinear relationship between expected and measured DNA concentrations. We have used statistical criteria to establish formal mathematical models for both quantitative and droplet digital PCR. To assess the method, a new Grass Carp ( Ctenopharyngodon idella ) TaqMan assay was developed and tested on both PCR platforms using eDNA in water samples. The LOD adjustment reduced Grass Carp occupancy and detection estimates while increasing uncertainty – indicating that caution needs to be applied to eDNA data without LOD correction. Compared to quantitative PCR, digital PCR had higher occurrence estimates due to increased sensitivity and dilution of inhibitors at low concentrations. Without accurate LOD correction, species occurrence and detection probabilities based on eDNA estimates are prone to a source of bias that cannot be reduced by an increase in sample size or PCR replicates. Other applications also could benefit from a standardized LOD such as GMO food analysis, and forensic and clinical diagnostics.

Molecular Ecology Resources

Phylogeographic implications for release of critically endangered manatee calves rescued in Northeast Brazil

1. The Antillean manatee (Trichechus manatus manatus), a subspecies of the West Indian manatee, is a large-bodied marine mammal found in fresh, brackish, and marine habitats throughout the Caribbean Islands and Central and South America. Antillean manatees in Brazil are classified as critically endangered, with a census size of approximately 500 individuals. The population in the Northeast region of Brazil is suspected to have approximately 300 manatees and is threatened by habitat alteration and incidental entanglement in fishing gear. 2. A high incidence of dependent calf strandings have been identified near areas of altered critical manatee habitat. The majority of the calves are neonates, discovered alive, with no potential mothers nearby. These calves typically require human intervention to survive. Since 1989 the calves have been rescued (N=67), rehabilitated, and released (N=25) to supplement the small wild manatee population. The rescued calves, and those born in captivity, are typically, not released to their rescue location, mainly for logistical reasons. Therefore, phylogeographic analyses can help to identify related populations and appropriate release sites. 3. Here, mitochondrial DNA analyses identified low haplotype (h=0.08) and nucleotide (π=0.0026) genetic diversity in three closely related haplotypes. All three haplotypes (M01, M03, and a previously unidentified haplotype, M04) were found in the northern portion of the region, while only a single haplotype (M01) was represented in the south. This suggests the presence of two genetic groups with a central mixing zone. Release of rehabilitated calves to unrelated populations may result in genetic swamping of locally adapted alleles or genotypes, limiting the evolutionary potential of the population. 4. The small population size coupled with low genetic diversity indicates that the Northeast Brazil manatee population is susceptible to inbreeding depression and possible local extinction. Further conservation measures incorporating genetic information could be beneficial to the critically endangered Brazilian manatee population.

Aquatic Conservation: Marine and Freshwater Ecosys