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Lakyn R. Sanders

Publications and source records attributed to Lakyn R. Sanders.

8 recordsLinked to original sources

Reconnaissance of surface water estrogenicity and the prevalence of intersex in smallmouth bass (Micropterus dolomieu) inhabiting New Jersey

The observation of testicular oocytes in male fishes has been utilized as a biomarker of estrogenic endocrine disruption. A reconnaissance project led in the Northeastern United States (US) during the period of 2008–2010 identified a high prevalence of intersex smallmouth bass on or near US Fish & Wildlife Service National Wildlife Refuges that included the observation of 100% prevalence in smallmouth bass males collected from the Wallkill River, NJ, USA. To better assess the prevalence of intersex smallmouth bass across the state of New Jersey, a tiered reconnaissance approach was initiated during the fall of 2016. Surface water samples were collected from 101 (85 river, 16 lake/reservoir) sites across the state at base-flow conditions for estrogenicity bioassay screening. Detectable estrogenicity was observed at 90% of the sites and 64% were above the US Environmental Protection Agency trigger level of 1 ng/L. Median surface water estrogenicity was 1.8 ng/L and a maximum of 6.9 ng/L E2EqBLYES was observed. Adult smallmouth bass were collected from nine sites, pre-spawn during the spring of 2017. Intersex was identified in fish at all sites, and the composite intersex prevalence was 93.8%. Prevalence across sites ranged from 70.6% to 100%. In addition to intersex, there was detectable plasma vitellogenin in males at all sites. Total estrogenicity in surface water was determined at these fish collection sites, and notable change over time was observed. Correlation analysis indicated significant positive correlations between land use (altered land; urban + agriculture) and surface water estrogenicity. There were no clear associations between land use and organismal metrics of estrogenic endocrine disruption (intersex or vitellogenin). This work establishes a baseline prevalence of intersex in male smallmouth bass in the state of New Jersey at a limited number of locations and identifies a number of waterbodies with estrogenic activity above an effects-based threshold.

New Jersey

Establishing molecular methods to quantitatively profile gastric diet items of fish—Application to the invasive blue catfish (ictalurus furcatus)

Understanding the diet of invasive species helps researchers to more accurately assess the health, survivorship, growth, and stability of an invasive fish species, as well as their effects on native populations. Techniques capable of identifying multiple prey species from fish stomach contents have been developed. In this study, a multi-locus metabarcoding approach was used to identify fish and invertebrate prey in stomach samples of Ictalurus furcatus (blue catfish), which were collected from two sites on the Mattawomen Creek and Nanjemoy Creek in Maryland. The mitochondrial 12S (mt12S) and mitochondrial 16S (mt16S) gene regions were sequenced and compared. First, a mock sample for each gene region was created with the pooled polymerase chain reaction product of known fish species, and quantities of the sample were used to determine efficacy of the amplicon. Results varied between gene regions analyzed. Then, when using the mt12S primers, next-generation sequencing determined that nine fish species were found at levels greater than 1 percent of the diet of blue catfish. The most common species were Perca flavescens (yellow perch) and Cyprinus carpio (common carp). The mt16S gene region analyses found 10 fish species at greater than 1 percent of the diet, which primarily included Orconectes limosus (spinycheek crayfish), Alosa pseudoharengus (alewife), and yellow perch. Partially digested eggs were identified using next-generation sequencing of yellow perch in two of the stomach samples, and a TaqMan® quantitative polymerase chain reaction (qPCR) assay was developed to more economically identify egg species in the future. The yellow-perch-specific TaqMan® qPCR assay was tested using primers that were developed to detect a 154-base-pair amplicon in the mitochondrial control region. Consumption of yellow perch eggs indicates that blue catfish could potentially negatively affect young-of-year recruitment of this native sportfish. Analyses of two gene regions helped confirm the major prey of the fish sampled and allowed identification of fish species as prey that were not included in a database for the two gene regions. We concluded that the mitochondrial ribosomal-marker-based next-generation sequencing method is useful in determining the prey of fish species.

Potomac River

Isolation, characterization and molecular identification of a novel aquareovirus that infects the endangered fountain darter, Etheostoma fonticola

The fountain darter Etheostoma fonticola (FOD) is a federally endangered fish listed under the US Endangered Species Act. Here, we identified and characterized a novel aquareovirus isolated from wild fountain darters inhabiting the San Marcos River. This virus was propagated in Chinook salmon embryo (CHSE)-214, rainbow trout gonad-2 and fathead minnow cells at 15°C. The epithelioma papulosum cyprini cell line was refractory at all temperatures evaluated. High throughput sequencing technologies facilitated the complete genome sequencing of this virus utilizing ribosomal RNA-depleted RNA extracted from infected CHSE-214 cells. Conventional PCR primer sets were developed for the detection and confirmation of this virus to assist diagnostic screening methods. Phylogenetic analysis suggests this virus belongs to the Aquareovirus A genus. This research provides requisite initial data critical to support hatchery and refugia biosecurity measures for this endangered species.

Diseases of Aquatic Organisms

A method for determining avian influenza virus hemagglutinin and neuraminidase subtype association

Methods for grouping specific avian influenza virus (AIV) hemagglutinin (HA) and neuraminidase (NA) subtype reverse-transcription polymerase chain reaction (RT-PCR) products into HA:NA subtypes when egg incubation is technically not feasible were evaluated. These approaches were adopted for use as post hoc methods after melt curve analysis. The methods are based on ratios obtained from amplicon copy count and amplicon molarity and were founded on the premise that infectious particles contain an equal copy count of single-stranded ribonucleic acid segments that encode HA or NA, and thus subtype-specific amplicons from a single AIV isolate should yield a theoretical HA:NA ratio of 1. Single and mixed HA:NA AIV subtype samples were evaluated to determine whether the calculated HA:NA ratios would approach the theoretical value. With these samples, preference was given to the molarity methods to better define and correct for the effects of multiple potential amplicons in the amplification mix. Further, the molarity method was used to evaluate pond sediment spiked with intact virus of known HA:NA subtype to determine whether the method is sufficiently robust to be used with complex samples, such as those acquired from waterfowl habitat. This was a proof-of-concept study intended to guide future methods development. The methods here are not meant to be applied in any other context. From the analysis of fully characterized isolates of North American AIV, the HA:NA molarity-based ratios were found to be 1.63 ± 0.75 (mean ± standard deviation) when corrected for the difference in amplification strength and the production of multiple amplicons in some reactions using equations developed in this study. Copy count HA:NA ratios, obtained from HA and NA subtype (RT-qPCR), were 1.146 ± 0.124 (mean ± standard deviation) when corrected for amplification efficiency. Correct associations of HA:NA subtype sample composition were made with mixed samples containing 1 HA and 2 NA, and 2 HA and 2 NA. When spiked pond sediment was evaluated, the molar ratio obtained for the H4 and N6 identified in the sample was 1.28 with correction and 1.14 without correction.

Open-File Report

An evaluation of the toxicity of potassium chloride, active compound in the molluscicide potash, on salmonid fish and their forage base

Potash, with the active ingredient potassium chloride (KCl) is a chemical that is currently being evaluated for potential use as a molluscicide to combat invasive zebra mussels and quagga mussels in Western United States waters. Although data available for other freshwater fishes indicate that recommended treatment levels of potash as a molluscicide are sublethal, this has not been demonstrated for all salmonid species. The objectives of this study were to perform toxicity testing to determine the lethality of potassium chloride against selected species of salmonid fish (brook trout and Chinook salmon) and selected invertebrate forage, and to identify any potential adverse physiological impacts of KCl to these salmonids in water at treatment levels used for mollusk eradication. Minimal mortality (n=1 fish) was observed during 96-hour toxicity testing at KCl concentrations of 0 to 800 milligrams per liter (mg/L), indicating that the lethal concentration (LC 50 ) values in these salmonid species were considerably higher than realistic molluscicide treatment concentrations. Sublethal effects were examined through evaluation of behavioral and morphological (histological) observation as well as specific blood chemistry parameters (electrolytes, osmolality, glucose, and cortisol). There was no strong evidence of significant physiological impairment among the two salmonid species due to KCl exposure. Whereas statistically significant differences in some parameters were observed in association with KCl treatments, it is unlikely that these differences indicate adverse biological impacts. Acute toxicity tests were conducted with invertebrate species at KCl exposure concentrations of 0–3,200 mg/L. Daphniid exposure trials resulted in differences in mortality among the test groups with higher mortality evident among the higher KCl exposure concentrations with a calculated LC 50 value of 196 mg/L KCl for a 48-hour exposure. Crayfish exposed to higher concentrations of KCl at or above 800 mg/L as specimens exhibited death or reversible paralysis. Chironomid larvae exposures were largely inconclusive because of cannibalistic behavior among the various test groups.

Open-File Report

Temporal evaluation of estrogenic endocrine disruption markers in smallmouth bass (Micropterus dolomieu) reveals seasonal variability in intersex

A reconnaissance project completed in 2009 identified intersex and elevated plasma vitellogenin in male smallmouth bass inhabiting the Missisquoi River, VT. In an attempt to identify the presence and seasonality of putative endocrine disrupting chemicals or other factors associated with these observations, a comprehensive reevaluation was conducted between September 2012 and June 2014. Here, we collected smallmouth bass from three physically partitioned reaches along the river to measure biomarkers of estrogenic endocrine disruption in smallmouth bass. In addition, polar organic chemical integrative samples (POCIS) were deployed to identify specific chemicals associated with biological observations. We did not observe biological differences across reaches indicating the absence of clear point source contributions to the observation of intersex. Interestingly, intersex prevalence and severity decreased in a stepwise manner over the timespan of the project. Intersex decreased from 92.8% to 28.1%. The only significant predictor of intersex prevalence was year of capture, based on logistic regression analysis. The mixed model of fish length and year-of-capture best predicted intersex severity. Intersex severity was also significantly different across late summer and early spring collections indicating seasonal changes in this metric. Plasma vitellogenin and liver vitellogenin Aa transcript abundance in males did not indicate exposure to estrogenic endocrine disrupting chemicals at any of the four sample collections. Analysis of chemicals captured by the POCIS as well as results of screening discrete water samples or POCIS extracts did not indicate the contribution of appreciable estrogenic chemicals. It is possible that unreported changes in land-use activity have ameliorated the problem, and our observations indicate recovery. Regardless, this work clearly emphasizes that single, snap shot sampling for intersex may not yield representative data given that the manifestation of this condition within a population can change dramatically over time.

Vermont

Spread of the Rat Lungworm (Angiostrongylus cantonensis) in Giant African Land Snails (Lissachatina fulica) in Florida, USA

The rat lungworm ( Angiostrongylus cantonensis ) is a parasitic nematode that causes rat lungworm disease. It is the leading cause of eosinophilic meningitis and is a zoonotic health risk. We confirmed the presence of A. cantonensis using species-specific, quantitative PCR in 18 of 50 (36%) giant African land snails ( Lissachatina fulica ) collected from Miami, Florida in May 2013. These snails were collected from seven of 21 core areas that the Florida Department of Agriculture and Consumer Services monitor weekly. Rat lungworms have not previously been identified in these areas. Duplicate DNA extractions of foot muscle tissue from each snail were tested. Of the seven core areas we examined, six were positive for A. cantonensis and prevalence of infection ranged from 27% to 100%. Of the 18 positive snails, only five were positive in both extractions. Our results confirm an increase in the range and prevalence of rat lungworm infection in Miami. We also emphasize the importance of extracting sufficient host tissue to minimize false negatives.

Florida