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Nancy D. Denslow

Publications and source records attributed to Nancy D. Denslow.

At least 19 recordsLinked to original sources

Systematic assessment of long-read RNA-seq methods for transcript identification and quantification

The Long-read RNA-Seq Genome Annotation Assessment Project Consortium was formed to evaluate the effectiveness of long-read approaches for transcriptome analysis. Using different protocols and sequencing platforms, the consortium generated over 427 million long-read sequences from complementary DNA and direct RNA datasets, encompassing human, mouse and manatee species. Developers utilized these data to address challenges in transcript isoform detection, quantification and de novo transcript detection. The study revealed that libraries with longer, more accurate sequences produce more accurate transcripts than those with increased read depth, whereas greater read depth improved quantification accuracy. In well-annotated genomes, tools based on reference sequences demonstrated the best performance. Incorporating additional orthogonal data and replicate samples is advised when aiming to detect rare and novel transcripts or using reference-free approaches. This collaborative study offers a benchmark for current practices and provides direction for future method development in transcriptome analysis.

Nature Methods

In vitro impacts of glyphosate on manatee lymphocytes

Exposure to contaminants, such as the herbicide glyphosate, can suppress protective immune functions. Glyphosate is the herbicide most used worldwide and has been found in the plasma of more than 50 % of the Florida manatees ( Trichechus manatus latirostris ) and all-year-round in their aquatic environment. Our objectives were to analyze the consequences of glyphosate exposure on their immune responses via T-lymphocyte proliferation assays and transcriptomics. We isolated peripheral blood mononuclear cells (mainly lymphocytes) of free-ranging manatees and performed T-cell proliferation assays. We used transcriptomics to understand the consequences of glyphosate in vitro exposure. The three doses chosen ranged from environmentally relevant concentrations at 10 to 10,000 µg.L −1 that is considered an environmental contamination scenario. Glyphosate caused a dose-dependent reduction in T-lymphocyte proliferation, with a significant mean reduction of 27.3 % at 10,000 µg.L −1 and up to 51.5 % in some individuals. Additionally, T-lymphocyte proliferation was significantly reduced in mid-winter compared to early winter. Transcriptomic analysis of peripheral blood mononuclear cells indicated that all doses of glyphosate (10, 1,000, and 10,000 µg.L −1 ) resulted in up-regulation of genes related to acute phase inflammation and inhibition of the T-lymphocyte proliferation pathway. Exposure to this contaminant along with other environmental stressors, such as extreme winters and red tide, might further affect the adaptive immune response of this threatened species.

Florida

Chronic exposure to glyphosate in Florida manatee

Florida manatees depend on freshwater environments as a source of drinking water and as warm-water refuges. These freshwater environments are in direct contact with human activities were glyphosate-based herbicides are being used. Glyphosate is the most used herbicide worldwide and it is intensively used in Florida as a sugarcane ripener and to control invasive aquatic plants. The objective of the present study was to determine the concentration of glyphosate and its breakdown product, aminomethylphosphonic acid (AMPA), in Florida manatee plasma and assess their exposure to manatees seeking a warm-water refuge in Crystal River (west central Florida), and in South Florida. We analyzed glyphosate’s and AMPA’s concentrations in Florida manatee plasma (n = 105) collected during 2009–2019 using HPLC-MS/MS. We sampled eight Florida water bodies between 2019 and 2020, three times a year: before, during and after the sugarcane harvest using grab samples and molecular imprinted passive Polar Organic Chemical Integrative Samplers (MIP-POCIS). Glyphosate was present in 55.8% of the sampled Florida manatees’ plasma. The concentration of glyphosate has significantly increased in Florida manatee samples from 2009 until 2019. Glyphosate and AMPA were ubiquitous in water bodies. The concentration of glyphosate and AMPA was higher in South Florida than in Crystal River, particularly before and during the sugarcane harvest when Florida manatees depend on warm water refuges. Based on our results, Florida manatees were chronically exposed to glyphosate and AMPA, during and beyond the glyphosate applications to sugarcane, possibly associated with multiple uses of glyphosate-based herbicides for other crops or to control aquatic weeds. This chronic exposure in Florida water bodies may have consequences for Florida manatees’ immune and renal systems which may further be compounded by other environmental exposures such as red tide or cold stress.

Florida

Investigating the gene expression profiles of rehabilitated Florida manatees (Trichechus manatus latirostris) following red tide exposure

To investigate a Florida manatee ( Trichechus manatus latirostris ) mortality event following a red tide bloom in Southwest Florida, an RNA sequencing experiment was conducted. Gene expression changes in white blood cells were assessed in manatees rescued from a red tide affected area (n = 4) and a control group (n = 7) using RNA sequencing. The genes with the largest fold changes were compared between the two groups to identify molecular pathways related to cellular and disease processes. In total, 591 genes (false discovery rate <0.05) were differentially expressed in the red tide group. Of these, 158 were upregulated and 433 were downregulated. This suggests major changes in white blood cell composition following an exposure to red tide. The most highly upregulated gene, Osteoclast associated 2C immunoglobulin-like receptor (OSCAR), was upregulated 12-fold. This gene is involved in initiating the immune response and maintaining a role in adaptive and innate immunity. The most highly downregulated gene, Piccolo presynaptic cytomatrix protein (PCLO), was downregulated by a factor of 977-fold. This gene is associated with cognitive functioning and neurotransmitter release. Downregulation of this gene in other studies was associated with neuronal loss and neuron synapse dysfunction. Among the cellular pathways that were most affected, immune response, including inflammation, wounds and injuries, cell proliferation, and apoptosis were the most predominant. The pathway with the most differentially expressed genes was the immune response pathway with 98 genes involved, many of them downregulated. Assessing the changes in gene expression associated with red tide exposure enhances our understanding of manatee immune response to the red tide toxins and will aid in the development of red tide biomarkers.

Florida

Review of and recommendations for monitoring contaminants and their effects in the San Francisco Bay−Delta

Legacy and current-use contaminants enter into and accumulate throughout the San Francisco Bay−Delta (Bay−Delta), and are present at concentrations with known effects on species important to this diverse watershed. There remains major uncertainty and a lack of focused research able to address and provide understanding of effects across multiple biological scales, despite previous and ongoing emphasis on the need for it. These needs are challenging specifically because of the established regulatory programs that often monitor on a chemical- by-chemical basis, or in which decisions are grounded in lethality-based endpoints. To best address issues of contaminants in the Bay−Delta, monitoring efforts should consider effects of environmentally relevant mixtures and sub- lethal impacts that can affect ecosystem health. These efforts need to consider the complex environment in the Bay−Delta including variable abiotic (e.g., temperature, salinity) and biotic (e.g., pathogens) factors. This calls for controlled and focused research, and the development of a multi-disciplinary contaminant monitoring and assessment program that provides information across biological scales. Information gained in this manner will contribute toward evaluating parameters that could alleviate ecologically detrimental outcomes. This review is a result of a Special Symposium convened at the University of California−Davis (UCD) on January 31, 2017 to address critical information needed on how contaminants affect the Bay−Delta. The UCD Symposium focused on new tools and approaches for assessing multiple stressor effects to freshwater and estuarine systems. Our approach is similar to the recently proposed framework laid out by the U.S. Environmental Protection Agency (USEPA) that uses weight of evidence to scale toxicological responses to chemical contaminants in a laboratory, and to guide the conservation of priority species and habitats. As such, we also aimed to recommend multiple endpoints that could be used to promote a multi-disciplinary understanding of contaminant risks in Bay−Delta while supporting management needs.

California

Recommended approaches to the scientific evaluation of ecotoxicological hazards and risks of endocrine-active substances

A SETAC Pellston Workshop ® “Environmental Hazard and Risk Assessment Approaches for Endocrine-Active Substances (EHRA)” was held in February 2016 in Pensacola, Florida, USA. The primary objective of the workshop was to provide advice, based on current scientific understanding, to regulators and policy makers; the aim being to make considered, informed decisions on whether to select an ecotoxicological hazard- or a risk-based approach for regulating a given endocrine-disrupting substance (EDS) under review. The workshop additionally considered recent developments in the identification of EDS. Case studies were undertaken on 6 endocrine-active substances (EAS—not necessarily proven EDS, but substances known to interact directly with the endocrine system) that are representative of a range of perturbations of the endocrine system and considered to be data rich in relevant information at multiple biological levels of organization for 1 or more ecologically relevant taxa. The substances selected were 17α-ethinylestradiol, perchlorate, propiconazole, 17β-trenbolone, tributyltin, and vinclozolin. The 6 case studies were not comprehensive safety evaluations but provided foundations for clarifying key issues and procedures that should be considered when assessing the ecotoxicological hazards and risks of EAS and EDS. The workshop also highlighted areas of scientific uncertainty, and made specific recommendations for research and methods-development to resolve some of the identified issues. The present paper provides broad guidance for scientists in regulatory authorities, industry, and academia on issues likely to arise during the ecotoxicological hazard and risk assessment of EAS and EDS. The primary conclusion of this paper, and of the SETAC Pellston Workshop on which it is based, is that if data on environmental exposure, effects on sensitive species and life-stages, delayed effects, and effects at low concentrations are robust, initiating environmental risk assessment of EDS is scientifically sound and sufficiently reliable and protective of the environment. In the absence of such data, assessment on the basis of hazard is scientifically justified until such time as relevant new information is available.

Integrated Environmental Assessment and Management

Current limitations and recommendations to improve testing for the environmental assessment of endocrine active substances

In the present study, existing regulatory frameworks and test systems for assessing potential endocrine active chemicals are described, and associated challenges are discussed, along with proposed approaches to address these challenges. Regulatory frameworks vary somewhat across geographies, but all basically evaluate whether a chemical possesses endocrine activity and whether this activity can result in adverse outcomes either to humans or to the environment. Current test systems include in silico, in vitro, and in vivo techniques focused on detecting potential endocrine activity, and in vivo tests that collect apical data to detect possible adverse effects. These test systems are currently designed to robustly assess endocrine activity and/or adverse effects in the estrogen, androgen, and thyroid hormone signaling pathways; however, there are some limitations of current test systems for evaluating endocrine hazard and risk. These limitations include a lack of certainty regarding: 1) adequately sensitive species and life stages; 2) mechanistic endpoints that are diagnostic for endocrine pathways of concern; and 3) the linkage between mechanistic responses and apical, adverse outcomes. Furthermore, some existing test methods are resource intensive with regard to time, cost, and use of animals. However, based on recent experiences, there are opportunities to improve approaches to and guidance for existing test methods and to reduce uncertainty. For example, in vitro high-throughput screening could be used to prioritize chemicals for testing and provide insights as to the most appropriate assays for characterizing hazard and risk. Other recommendations include adding endpoints for elucidating connections between mechanistic effects and adverse outcomes, identifying potentially sensitive taxa for which test methods currently do not exist, and addressing key endocrine pathways of possible concern in addition to those associated with estrogen, androgen, and thyroid signaling.

Integrated Environmental Assessment and Management

Potential estrogenic effects of wastewaters on gene expression in Pimephales promelas and fish assemblages in streams of southeastern New York

Direct linkages between endocrine-disrupting compounds (EDCs) from municipal and industrial wastewaters and impacts on wild fish assemblages are rare. The levels of plasma vitellogenin (Vtg) and Vtg messenger ribonucleic acid (mRNA) in male fathead minnows (Pimephales promelas) exposed to wastewater effluents and dilutions of 17&alpha;-ethinylestradiol (EE2), estrogen activity, and fish assemblages in 10 receiving streams were assessed to improve understanding of important interrelations. Results from 4-d laboratory assays indicate that EE2, plasma Vtg concentration, and Vtg gene expression in fathead minnows, and 17&beta;-estradiol equivalents (E2Eq values) were highly related to each other (R2&thinsp;=&thinsp;0.98&ndash;1.00). Concentrations of E2Eq in most effluents did not exceed 2.0&thinsp;ng/L, which was possibly a short-term exposure threshold for Vtg gene expression in male fathead minnows. Plasma Vtg in fathead minnows only increased significantly (up to 1136&thinsp;&mu;g/mL) in 2 wastewater effluents. Fish assemblages were generally unaffected at 8 of 10 study sites, yet the density and biomass of 79% to 89% of species populations were reduced (63&ndash;68% were reduced significantly) in the downstream reach of 1 receiving stream. These results, and moderate to high E2Eq concentrations (up to 16.1&thinsp;ng/L) observed in effluents during a companion study, suggest that estrogenic wastewaters can potentially affect individual fish, their populations, and entire fish communities in comparable systems across New York, USA.

New York

Transcriptomic effects-based monitoring for endocrine active chemicals: Assessing relative contribution of treated wastewater to downstream pollution

The present study investigated whether a combination of targeted analytical chemistry information with unsupervised, data-rich biological methodology (i.e., transcriptomics) could be utilized to evaluate relative contributions of wastewater treatment plant (WWTP) effluents to biological effects. The effects of WWTP effluents on fish exposed to ambient, receiving waters were studied at three locations with distinct WWTP and watershed characteristics. At each location, 4 d exposures of male fathead minnows to the WWTP effluent and upstream and downstream ambient waters were conducted. Transcriptomic analyses were performed on livers using 15 000 feature microarrays, followed by a canonical pathway and gene set enrichment analyses. Enrichment of gene sets indicative of teleost brain–pituitary–gonadal–hepatic (BPGH) axis function indicated that WWTPs serve as an important source of endocrine active chemicals (EACs) that affect the BPGH axis (e.g., cholesterol and steroid metabolism were altered). The results indicated that transcriptomics may even pinpoint pertinent adverse outcomes (i.e., liver vacuolization) and groups of chemicals that preselected chemical analytes may miss. Transcriptomic Effects-Based monitoring was capable of distinguishing sites, and it reflected chemical pollution gradients, thus holding promise for assessment of relative contributions of point sources to pollution and the efficacy of pollution remediation.

Environmental Science & Technology

Correlation of gene expression and contaminat concentrations in wild largescale suckers: a field-based study

Toxic compounds such as organochlorine pesticides (OCs), polychlorinated biphenyls (PCBs), and polybrominated diphenyl ether flame retardants (PBDEs) have been detected in fish, birds, and aquatic mammals that live in the Columbia River or use food resources from within the river. We developed a custom microarray for largescale suckers (Catostomus macrocheilus) and used it to investigate the molecular effects of contaminant exposure on wild fish in the Columbia River. Using Significance Analysis of Microarrays (SAM) we identified 72 probes representing 69 unique genes with expression patterns that correlated with hepatic tissue levels of OCs, PCBs, or PBDEs. These genes were involved in many biological processes previously shown to respond to contaminant exposure, including drug and lipid metabolism, apoptosis, cellular transport, oxidative stress, and cellular chaperone function. The relation between gene expression and contaminant concentration suggests that these genes may respond to environmental contaminant exposure and are promising candidates for further field and laboratory studies to develop biomarkers for monitoring exposure of wild fish to contaminant mixtures found in the Columbia River Basin. The array developed in this study could also be a useful tool for studies involving endangered sucker species and other sucker species used in contaminant research.

Oregon;Washington

Methylmercury-induced changes in gene transcription associated with neuroendocrine disruption in largemouth bass (Micropterus salmoides)

Methyl-mercury (MeHg) is a potent neuroendocrine disruptor that impairs reproductive processes in fish. The objectives of this study were to (1) characterize transcriptomic changes induced by MeHg exposure in the female largemouth bass (LMB) hypothalamus under controlled laboratory conditions, (2) investigate the health and reproductive impacts of MeHg exposure on male and female largemouth bass (LMB) in the natural environment, and (3) identify MeHg-associated gene expression patterns in whole brain of female LMB from MeHg-contaminated habitats. The laboratory experiment was a single injection of 2.5 μg MeHg/g body weight for 96 h exposure. The field survey compared river systems in Florida, USA with comparably lower concentrations of MeHg (Wekiva, Santa Fe, and St. Johns Rivers) in fish and one river system with LMB that contained elevated concentrations of MeHg (St. Marys River). Microarray analysis was used to quantify transcriptomic responses to MeHg exposure. Although fish at the high-MeHg site did not show overt health or reproductive impairment, there were MeHg-responsive genes and pathways identified in the laboratory study that were also altered in fish from the high-MeHg site relative to fish at the low-MeHg sites. Gene network analysis suggested that MeHg regulated the expression targets of neuropeptide receptor and steroid signaling, as well as structural components of the cell. Disease-associated gene networks related to MeHg exposure, based upon expression data, included cerebellum ataxia, movement disorders, and hypercalcemia. Gene responses in the CNS are consistent with the documented neurotoxicological and neuroendocrine disrupting effects of MeHg in vertebrates.

General and Comparative Endocrinology

Custom microarray construction and analysis for determining potential biomarkers of subchronic androgen exposure in the Eastern Mosquitofish ( Gambusia holbrooki )

Background The eastern mosquitofish ( Gambusia holbrooki ) has the potential to become a bioindicator organism of endocrine disrupting chemicals (EDCs) due to its androgen-driven secondary sexual characteristics. However, the lack of molecular information on G . holbrooki hinders its use as a bioindicator coupled with biomarker data. While traditional gene-by-gene approaches provide insight for biomarker development, a holistic analysis would provide more rapid and expansive determination of potential biomarkers. The objective of this study was to develop and utilize a mosquitofish microarray to determine potential biomarkers of subchronic androgen exposure. To achieve this objective, two specific aims were developed: 1) Sequence a G . holbrooki cDNA library, and 2) Use microarray analysis to determine genes that are differentially regulated by subchronic androgen exposure in hepatic tissues of 17&beta;-trenbolone (TB) exposed adult female G . holbrooki . Results A normalized library of multiple organs of male and female G . holbrooki was prepared and sequenced by the Illumina GA IIx and Roche 454 XLR70. Over 30,000 genes with e-value&thinsp;&le;&thinsp;10 -4 were annotated and 14,758 of these genes were selected for inclusion on the microarray. Hepatic microarray analysis of adult female G . holbrooki exposed to the vehicle control or 1 &mu;g/L of TB (a potent anabolic androgen) revealed 229 genes upregulated and 279 downregulated by TB (one-way ANOVA, p&thinsp;<&thinsp;0.05, FDR &alpha;&thinsp;=&thinsp;0.05, fold change&thinsp;>&thinsp;1.5 and&thinsp;<&thinsp;&minus;1.5). Fifteen gene ontology biological processes were enriched by TB exposure (Fisher&rsquo;s Exact Test, p&thinsp;<&thinsp;0.05). The expression levels of 17&beta; - hydroxysteroid dehydrogenase 3 and zona pellucida glycoprotein 2 were validated by quantitative polymerase chain reaction (qPCR) (Student&rsquo;s t-test, p&thinsp;<&thinsp;0.05). Conclusions Coupling microarray data with phenotypic changes driven by androgen exposure in mosquitofish is key for developing this organism into a bioindicator for EDCs. Future studies using this array will enhance knowledge of the biology and toxicological response of this species. This work provides a foundation of molecular knowledge and tools that can be used to delve further into understanding the biology of G . holbrooki and how this organism can be used as a bioindicator organism for endocrine disrupting pollutants in the environment.

BMC Genomics

Characterization of plasma vitellogenin and sex hormone concentrations during the annual reproductive cycle of the endangered razorback sucker

Population declines of the endangered razorback sucker Xyrauchen texanus in the Colorado River basin have been attributed to predation by and competition with nonnative fishes, habitat alteration, and dam construction. The reproductive health and seasonal variation of the reproductive end points of razorback sucker populations are currently unknown. Using nonlethal methods, we characterized the plasma hormonal fluctuations of reproductively mature female and male razorback suckers over a 12-month period in a hatchery by measuring their vitellogenin (VTG) and three sex hormones: 17&beta;-estradiol (E2), testosterone (T), and 11-ketotestosterone (KT). Fish were identified as reproductive or nonreproductive based on their body weight, VTG, and sex hormone profiles. In reproductive females, the E2 concentration increased in the fall and winter, and increases in T and VTG concentrations were generally associated with the spawning period. Mean T concentrations were consistently greater in reproductive females than in nonreproductive females, but this pattern was even more pronounced during the spawning period (spring). Consistently low T concentrations (<3 ng/mL) in adult females during the spawning period may indicate reproductive impairment. In reproductive males, spring increases in KT and T concentrations were associated with spawning; concentrations of E2 (<0.48 ng/mL) and VTG (<0.001 mg/mL) were low in males throughout the study. In addition, the E2 : KT ratio and T were the best metrics by which to distinguish female from male adult razorback suckers throughout the year. These metrics of reproductive health and condition may be particularly important to recovery efforts of razorback suckers given that the few remaining wild populations are located in a river where water quality and quantity issues are well documented. In addition to the size, age, and recruitment information currently considered in the recovery goals of this endangered species, reproductive end points could be included as recovery metrics with which to monitor seasonal trends and determine whether repatriated populations are cycling naturally.

North American Journal of Fisheries Management

Gene expression changes in female zebrafish (Danio rerio) brain in response to acute exposure to methylmercury

Methylmercury (MeHg) is a potent neurotoxicant and endocrine disruptor that accumulates in aquatic systems. Previous studies have shown suppression of hormone levels in both male and female fish, suggesting effects on gonadotropin regulation in the brain. The gene expression profile in adult female zebrafish whole brain induced by acute (96 h) MeHg exposure was investigated. Fish were exposed by injection to 0 or 0.5 µg MeHg/g. Gene expression changes in the brain were examined using a 22,000‐feature zebrafish microarray. At a significance level of p < 0.01, 79 genes were up‐regulated and 76 genes were down‐regulated in response to MeHg exposure. Individual genes exhibiting altered expression in response to MeHg exposure implicate effects on glutathione metabolism in the mechanism of MeHg neurotoxicity. Gene ontology (GO) terms significantly enriched among altered genes included protein folding, cell redox homeostasis, and steroid biosynthetic process. The most affected biological functions were related to nervous system development and function, as well as lipid metabolism and molecular transport. These results support the involvement of oxidative stress and effects on protein structure in the mechanism of action of MeHg in the female brain. Future studies will compare the gene expression profile induced in response to MeHg with that induced by other toxicants and will investigate responsive genes as potential biomarkers of MeHg exposure.

Environmental Toxicology and Chemistry

Organic Chemical Concentrations and Reproductive Biomarkers in Common Carp (Cyprinus carpio) Collected from Two Areas in Lake Mead, Nevada, May 1999-May 2000

The U.S. Geological Survey, in cooperation with the U.S. Fish and Wildlife Service, National Park Service, Bureau of Reclamation, and Nevada Department of Wildlife, collected and assessed data to determine the general health and reproductive status of common carp (Cyprinus carpio) at two study areas in Lake Mead, Nevada, during May 1999-May 2000. These data will form the basis of interpretations and provide a comparison for continuing studies on the health of the ecosystem in Lake Mead. One study area, Las Vegas Bay, is in the western part of Lake Mead. Las Vegas Bay receives inflows from Las Vegas Wash, which is predominantly tertiary-treated wastewater effluent, and to a lesser extent stormwater runoff from Las Vegas, Henderson, and other nearby communities, and from ground water underlying Las Vegas Valley. The other study area, Overton Arm, is in the northern extent of Lake Mead. Overton Arm receives inflow from the Virgin and Muddy Rivers, which historically are not influenced by wastewater effluent. Both sexes of common carp were collected bimonthly for 12 months using boat-mounted electrofishing gear (a direct electric current is used to temporarily immobilize fish for capture) to determine their health and reproductive status and any relation between these factors and environmental contaminants. This report presents fish tissue chemistry, organic chemical compound concentrations, and biomarker data for 83 male common carp collected from Las Vegas Bay, similar organic chemistry results for 15 male common carp, and similar biomarker measures for 80 male common carp collected from Overton Arm. Tissue chemistry results also are presented for 16 female common carp and biomarker measures for 79 female common carp collected from Las Vegas Bay, and tissue chemistry results for 15 female common carp and biomarker measures for 81 female common carp collected from Overton Arm. Thirty-three organic chemical compounds plus total concentrations for four groups of compounds (chlordanes, polychlorinated biphenyls [PCBs], brominated diphenyl ethers [BDEs], and triclosans) were analyzed from extracts of whole-body tissue using gas chromatography/mass spectrometry in male common carp from Las Vegas Bay during May 1999 through May 2000. All 33 compounds were detected in at least one sample of whole-body tissue from male common carp collected in Las Vegas Bay. In Overton Arm, 37 organic compounds plus total concentrations of three groups of compounds (PCBs, BDEs, and triclosans) were analyzed in male common carp where 20 (54 percent) of the compounds were detected. Sixteen of the 33 compounds detected in male common carp from Las Vegas Bay and 10 compounds detected in males from Overton Arm have the potential to disrupt the endocrine system in fish in Lake Mead. During May and June 1999, the mean concentration of all organic compounds detected in male common carp was 670 micrograms per kilogram from Las Vegas Bay and 109 micrograms per kilogram from Overton Arm. Twenty-seven organic compounds plus total PCBs were analyzed from extracts of whole-body tissue in female common carp collected in Las Vegas Bay and Overton Arm during May 1999. Twenty-four (86 percent) of these compounds were detected in at least one sample of whole-body tissue from female common carp collected from Las Vegas Bay while 10 (36 percent) chemical compounds were detected in female common carp from Overton Arm during that same period. Median concentrations of all chemical compounds were higher in female common carp from Las Vegas Bay compared to those collected from Overton Arm except Dacthal (DCPA), which was similar between sites. Biomarker measures obtained for male and female common carp include gonadosomatic index (percentage of gonad weight to total body weight), plasma vitellogenin (a phospholipid protein normally produced by female common carp and other oviparous fish), and condition factor [body weight/(fork length)3]. Biomarker measures for male c

Data Series

Biomonitoring of Environmental Status and Trends (BEST) Program: Environmental contaminants, health indicators, and reproductive biomarkers in fish from the Mobile, Apalachicola-Chattahoochee-Flint, Savannah, and Pee Dee River Basins

Largemouth bass ( Micropterus salmoides ) and common carp ( Cyprinus carpio ) were collected from 13 sites in 4 river basins in the southeastern United States to document spatial trends in accumulative contaminants, health indicators, and reproductive biomarkers. Organochlorine residues, 2,3,7,8- tetrachlorodibenzo-p-dioxin-like activity (TCDD-EQ), and elemental contaminants were measured in composite samples of whole fish, grouped by species and gender, from each site. Fish were field-examined for external and internal anomalies, selected organs were weighed to compute somatic indices, and tissue and fluid samples were preserved for fish health and reproductive biomarker analyses. Mercury concentrations in bass samples from all sites exceeded toxicity thresholds for mammals [>0.1 micrograms per gram wet weight (ug/g ww)], fish (>0.2 ug/g ww), and birds (>0.3 ug/g ww) and were greatest (>0.5 ug/g ww) in samples from the Alabama River at Eureka Landing, Alabama; the Mobile River at Bucks, Alabama; the Apalachicola River at Blountstown, Florida; the Savannah River at Sylvania, Georgia; and the Pee Dee River at Bucksport, South Carolina. Selenium concentrations were relatively high (>0.75 ug/g ww) in fish from the Tombigbee River at Lavaca, Alabama; the Mobile River at Bucks; and the Chattahoochee River at Omaha, Georgia compared to those from other sites. Concentrations of 2,2-bis (p-chlorophenyl)- 1,1-dichloroethylene (p,p'-DDE) were high in fish from the Chattahoochee River at Omaha and the Mobile River near Bucks, which was near a 2,2-bis (p-chlorophenyl)-1,1- dichloroethylene (DDT) formulating facility that historically discharged into the lower Mobile River. Toxaphene concentrations in fish from the Flint River near Albany, Georgia (60-100 nanograms per gram (ng/g) ww) may pose a risk to fish. Concentrations of other formerly used (total chlordanes, dieldrin, endrin, aldrin, mirex, and hexachlorobenzene) and currently used (pentachlorobenzene, pentachloroanisole, dacthal, endosulfan, y-HCH, and methoxychlor) organochlorine residues generally were low or did not exceed toxicity thresholds. Total polychlorinated biphenyls concentrations in samples from the Coosa River at Childersburg, Alabama; the Apalachicola River at Omaha; the Apalachicola River at Blountstown; and the Pee Dee River at Bucksport were >480 ng/g ww and may be a risk to piscivorous wildlife. Dioxin-like activity as measured by TCDD-EQ was greatest [>10 picograms per gram (pg/g)] in male fish from the Coosa River at Childersburg and the Mobile River at Bucks. Hepatic ethoxyresorufin O-deethylase activity generally was greatest in carp from the Mobile River Basin [means >10 picomols per minute per milligram of protein (pmol/min/mg)] and in bass from the Tombigbee River at Lavaca and Pee Dee River at Pee Dee, South Carolina (means >65 pmol/min/mg). Altered biomarkers were noted in fish from all basins. The field necropsy and histopathological examination determined that fish from the Mobile River Basin generally were in poorer health than those from the other basins. In bass, health assessment index scores were correlated with mercury and p,p'-DDE concentrations. High health assessment index scores in Mobile River Basin fish were widespread and caused primarily by parasitic infestations, which were most severe in fish from the Tombigbee River at Lavaca and the Alabama River at Eureka Landing. Tumors were present in few fish (n = 5; 0.01%). Ovarian tumors of the same origin (smooth muscle) were present in two older carp from the Chattahoochee River near Omaha, Georgia and may be contaminant related. Reproductive biomarkers including gonadosomatic index, vitellogenin concentrations, and steroid hormone concentrations were anomalous in fish from various sites but were not consistently related to any particular chemical contaminant. Intersex gonads were identified in 47 male bass (42%) representing 12 sites and may indicate exposure to endocrine disrupting comp

Mobile, Apalachicola-Chattahoochee-Flint, Savannah

Toxicogenomics in regulatory ecotoxicology

Recently, we have witnessed an explosion of different genomic approaches that, through a combination of advanced biological, instrumental, and bioinformatic techniques, can yield a previously unparalleled amount of data concerning the molecular and biochemical status of organisms. Fueled partially by large, well-publicized efforts such as the Human Genome Project, genomic research has become a rapidly growing topical area in multiple biological disciplines. Since 1999, when the term “toxicogenomics” was coined to describe the application of genomics to toxicology (1), a rapid increase in publications on the topic has occurred (Figure 1). The potential utility of toxicogenomics in toxicological research and regulatory activities has been the subject of scientific discussions and, as with any new technology, has evoked a wide range of opinion (2–6).

Environmental Science & Technology

Biomonitoring of Environmental Status and Trends (BEST) Program: Environmental contaminants, health indicators, and reproductive biomarkers in fish from the Colorado River basin

Seven fish species were collected from 14 sites on rivers in the Colorado River Basin (CDRB) from August to October 2003. Spatial trends in the concentrations of accumulative contaminants were documented and contaminant effects on the fish were assessed. Sites were located on the mainstem of the Colorado River and on the Yampa, Green, Gunnison, San Juan, and Gila Rivers. Common carp (Cyprinus carpio), black bass (Micropterus sp.), and channel catfish (Ictalurus punctatus) were the targeted species. Fish were field-examined for external and internal anomalies, selected organs were weighed to compute somatic indices, and tissue and fluid samples were preserved for fish health and reproductive biomarker analyses. Composite samples of whole fish, grouped by species and gender, from each site were analyzed for organochlorine and elemental contaminants using performance-based and instrumental methods. 2,3,7,8-tetrachlorodibenzo-p-dioxin-like activity (TCDD-EQ) was measured using the H4IIE rat hepatoma cell bioassay. Selenium (Se) and mercury (Hg) concentrations were elevated throughout the CDRB, and pesticides concentrations were greatest in fish from agricultural areas in the Lower Colorado River and Gila River. Selenium concentrations exceeded toxicity thresholds for fish (>1.0 ?g/g ww) at all sites except from the Gila River at Hayden, Arizona. Mercury concentrations were elevated (>0.1 ?g/g ww) in fish from the Yampa River at Lay, Colorado; the Green River at Ouray National Wildlife Refuge (NWR), Utah and San Rafael, Utah; the San Juan River at Hogback Diversion, New Mexico; and the Colorado River at Gold Bar Canyon, Utah, Needles, California, and Imperial Dam, Arizona. Concentrations of p,p'-DDE were relatively high in fish from Arlington, Arizona (>1.0 ?g/g ww) and Phoenix, Arizona (>0.5 ?g/g ww). Concentrations of other banned pesticides including toxaphene, total chlordanes, and dieldrin were also greatest at these two sites but did not exceed toxicity thresholds. Current-use or unlisted pesticides such as dacthal, endosulfan, '-HCH, and methoxychlor were also greatest in fish from Gila River. Total polychlorinated biphenyls (PCBs; >0.11 ?g/g ww) and TCDD-EQs (>5 pg/g ww) exceeded wildlife guidelines in fish from the Gila River at Phoenix, Arizona. Hepatic ethoxyresorufin O-deethylase (EROD) activity was also relatively high in carp from the Gila River at Phoenix, Arizona and in bass from the Green River at Ouray NWR, Utah. Altered biomarkers were noted in fish throughout the CDRB. Fish from some stations responded to chronic contaminant exposure as indicated by fish health and reproductive biomarker results. Multiple fish health indicators including altered body and organ weights and high health assessment index scores may be associated with elevated Se concentrations in fish from the Colorado River at Loma, Colorado and Needles, California. Although grossly visible external or internal lesions were found on most fish from some sites, histopathological analysis determined many of these to be inflammatory responses associated with parasites. Edema, exophthalmos, and cataracts were noted in fish from sites with elevated Se concentrations. Reproductive biomarkers including gonad development and maturation, vitellogenin concentrations, and steroid hormone concentrations were anomalous in fish from the Gila River at Hayden and Phoenix, Arizona. In addition, intersex fish were found at seven of 14 sites. The intersex condition was identified in smallmouth bass (M. dolomieu), largemouth bass (M. salmoides), channel catfish, and carp and may indicate exposure to endocrine disrupting compounds. Seven of ten male smallmouth bass from the Yampa River at Lay, Colorado were intersex. Male carp, bass, and channel catfish with low concentrations of vitellogenin were common in the CDRB. Comparatively high vitellogenin concentrations (>0.2 mg/mL) were measured in male fish from the Green River at Ouray NWR, Utah and the Colorado River at Im

Scientific Investigations Report