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Tumor Prevalence and Biomarkers of Exposure and Response in Brown Bullheads (Ameiurus nebulosus) from the Tidal Potomac River Watershed

Four groups of thirty brown bullheads (Ameiurus nebulosus) were collected from the tidal Potomac River watershed to survey tumor prevalence in relation to contaminant exposure. Fish were obtained from the Quantico embayment, near a Superfund site that released polychlorinated biphenyls (PCBs) and DDT compounds; Neabsco Creek,.a tributary with petroleum inputs from upstream areas and marinas; and the Anacostia River (both in spring and fall),where sediment is contaminated with polynuclear aromatic hydrocarbons (PAHs), PCBs, and organochlorine pesticides. Fish were also collected from the Tuckahoe River, on the Eastern Shore of Maryland, as a reference. Fish were necropsied and examined grossly and histopathologically for skin and liver neoplasms. Cytochrome P450 activity, bile PAH metabolites, and muscle organochlorine pesticide/PCB concentrations were determined in randomly selected individuals. There were significant differences among sites in liver tumor prevalence: Anacostia (spring)-50%, Anacostia (fall)-60%, Neabsco-17%, Quantico-7%, Tuckahoe-10%. Skin tumor prevalences were also significantly different: Anacostia (spring)-37%, Anacostia (fall)-10%, Neabsco-3%, Quantico-3%, Tuckahoe-0%. Tumor prevalences in Anacostia fish were comparable to those at contaminated sites in the Great Lakes. PAH concentrations were higher in Anacostia sediments than at the other sites and there were significantly higher concentrations of PAH metabolites in bile of the Anacostia fish. At present, there are insufficient data, however, to establish a cause-effect linkage with a particular class of contaminants. Tumor surveys in selected species are recommended for monitoring the status and remediation of Regions of Concern and other areas in the Chesapeake Bay watershed.

Report

Tumor prevalence and biomarkers in brown bullheads (Ameiurus nebulosus) from the tidal Potomac River watershed [poster]

Associations between contaminant exposure and liver and skin tumor prevalence were evaluated in brown bullheads (Ameiurus nebulosus) from the tidal Potomac River watershed. Thirty bullheads (>260 mm) were collected from (1) Quantico embayment, near a Superfund site which released organochlorine contaminants; (2) Neabsco Creek, a tributary with petroleum inputs from runoff and marinas; and (3) Anacostia River (spring and fall), an urban tributary designated as a Chesapeake Bay Region of Concern, contaminated with PCBs, PAHs, and organochlorine pesticides. Fish were collected from the Tuckahoe River, as a reference. Cytochrome P450 activity, bile PAH metabolites, and muscle organochlorine pesticide/PCB concentrations were measured in randomly-selected individuals and sediment contaminants were analyzed. We found statistically significant differences in liver tumor prevalences: Anacostia (spring)-50%, Anacostia (fall)-60%, Neabsco-17%, Quantico-7%, Tuckahoe-10%. Skin tumor prevalences were significantly different: Anacostia (spring)-37%, Anacostia (fall)-10%, Neabsco-3%, Quantico-3%, Tuckahoe-0%. Tumor prevalence in Anacostia bullheads warrants concern and was similar to those at highly contaminated sites in the Great Lakes. There was evidence of higher PAH exposure in Anacostia fish but a cause-effect linkage cannot be established. Fish tumor surveys, with histopathological examination of internal and external organs, are recommended for monitoring the status of Regions of Concern.

Society of Environmental Toxicology and Chemistry,

Tumor prevalence and biomarkers in brown bullheads (Ameiurus nebulosus) from the tidal Potomac River watershed [poster]

Associations between contaminant exposure and liver and skin tumor prevalence were evaluated in brown bullheads (Ameiurus nebulosus) from the tidal Potomac River watershed. Thirty bullheads (>260 mm) were collected from (1) Quantico embayment, near a Superfund site which released organochlorine contaminants; (2) Neabsco Creek, a tributary with petroleum inputs from runoff and marinas; and (3) Anacostia River (spring and fall), an urban tributary designated as a Chesapeake Bay Region of Concern, contaminated with PCBs, PAHs, and organochlorine pesticides. Fish were collected from the Tuckahoe River, as a reference. Cytochrome P450 activity, bile PAH metabolites, and muscle organochlorine pesticide/PCB concentrations were measured in randomly-selected individuals and sediment contaminants were analyzed. We found statistically significant differences in liver tumor prevalences: Anacostia (spring)-50%, Anacostia (fall)-60%, Neabsco-17%, Quantico-7%, Tuckahoe-10%. Skin tumor prevalences were significantly different: Anacostia (spring)-37%, Anacostia (fall)-10%, Neabsco-3%, Quantico-3%, Tuckahoe-0%. Tumor prevalence in Anacostia bullheads warrants concern and was similar to those at highly contaminated sites in the Great Lakes. There was evidence of higher PAH exposure in Anacostia fish but a cause-effect linkage cannot be established. Fish tumor surveys, with histopathological examination of internal and external organs, are recommended for monitoring the status of Regions of Concern.

Society of Environmental Toxicology and Chemistry,

Dietary versus nondietary fatty acid profiles of lake trout ecotypes from Lake Superior and Great Bear Lake: Are fish really what they eat?

Fatty acids are well-established biomarkers used to characterize trophic ecology, food-web linkages, and the ecological niche of many different taxa. Most often, fatty acids that are examined include only those previously identified as “dietary” or “extended dietary” biomarkers. Fatty acids considered as nondietary biomarkers, however, represent numerous fatty acids that can be extracted. Some studies may include nondietary fatty acids (i.e., combined with dietary fatty acids), but do not specifically assess them, whereas in other studies, these data are discarded. In this study, we explored whether nondietary biomarker fatty acids can provide worthwhile information by assessing their ability to discriminate intraspecific diversity within and between lakes. Nondietary fatty acids used as biomarkers delineated variation among regions, among locations within a lake, and among ecotypes within a species. Physiological differences that arise from differences in energy processing can be adaptive and linked to habitat use by a species’ ecotype and likely explains why nondietary fatty acid biomarkers can be a relevant tool to delineate intraspecific diversity. Little is known about the nondietary-mediated differences in fatty acid composition, but our results showed that nondietary fatty acid biomarkers can be useful tool in identifying variation.

Lake Superior

Davis Pond freshwater prediversion biomonitoring study: freshwater fisheries and eagles

In January 2001, the construction of the Davis Pond freshwater diversion structure was completed by the U.S. Army Corps of Engineers. The diversion of freshwater from the Mississippi River is intended to mitigate saltwater intrusion from the Gulf of Mexico and to lessen the concomitant loss of wetland areas. In addition to the freshwater inflow, Barataria Bay basin would receive nutrients, increased flows of sediments, and water-borne and sediment-bound compounds. The purpose of this biomonitoring study was, therefore, to serve as a baseline for prediversion concentrations of selected contaminants in bald eagle ( Haliaeetus leucocephalus ) nestlings (hereafter referred to as eaglets), representative freshwater fish, and bivalves. Samples were collected from January through June 2001. Two similarly designed postdiversion studies, as described in the biological monitoring program, are planned. Active bald eagle nests targeted for sampling eaglet blood (n = 6) were generally located southwest and south of the diversion structure. The designated sites for aquatic animal sampling were at Lake Salvador, at Lake Cataouatche, at Bayou Couba, and along the Mississippi River. Aquatic animals representative of eagle prey were collected. Fish were from three different trophic levels and have varying feeding strategies and life histories. These included herbivorous striped mullet ( Mugil cephalus ), omnivorous blue catfish ( Ictalurus furcatus ), and carnivorous largemouth bass ( Micropterus salmoides ). Three individuals per species were collected at each of the four sampling sites. Freshwater Atlantic rangia clams ( Rangia cuneata ) were collected at the downstream marsh sites, and zebra mussels ( Dreissena spp.) were collected on the Mississippi River. The U.S. Geological Survey (USGS) Biomonitoring of Environmental Status and Trends (BEST) protocols served as guides for fish sampling and health assessments. Fish are useful for monitoring aquatic ecosystems because they accumulate pesticides and other contaminants. Biomarker data on individual fish, generated at the USGS National Wetlands Research Center (Lafayette, La.), included percent white blood cells in whole blood, spleen weight to body weight ratio, liver weight to body weight ratio, condition factor, splenic macrophage aggregates, and liver microsomal 7-ethoxyresorufin-o-deethylase (EROD) activity. Fish age was estimated by comparing total lengths with values from the same species in the Southeast United States as determined from the literature. Contaminant analyses were coordinated by the U.S. Fish and Wildlife Service (USFWS) Analytical Control Facility (Laurel, Md.), where residues of organochlorine (OC) pesticides, total polychlorinated biphenyls (PCBs), polycyclic aromatic hydrocarbons (PAHs), aliphatic hydrocarbons (AHs), and trace elements were determined. The organic contaminant data were generated at the Mississippi State University Chemical Lab (Mississippi State, Miss.), and the inorganic contaminant data were generated by the Texas A&M University Geochemical and Environmental Research Group (College Station, Tex.). Statistical tests were performed to assess relationships among contaminants, fish age, fish species, and collection sites. Trends in interspecific differences among fish in concentrations of contaminants were noted. Striped mullet (hereafter mullet) frequently displayed the highest chemical concentrations. Levels of contaminants were generally higher in samples obtained from the Mississippi River than in those collected from the diversion area and were higher in mussels and clams (hereafter bivalves) than in fish. Because the Mississippi River sampling site for mullet and largemouth bass was downriver of the structure and south of New Orleans and the catfish site was upriver, the downriver data may not be directly reflective of the results from the receiving waters at the Davis Pond structure. Compared to the Caernarvon freshwater prediversion study in 1990 that assessed possible influx of contaminants with the freshwater diversion, contaminant levels in fishes and bivalves in this study were generally lower, yet three nontoxic inorganic elements in Davis Pond fish samples exhibited ranges of concentrations that were more than two times higher than did those from Caernarvon. Levels in bivalves were different between diversions but about equal in the numbers of trace elements showing high levels per location. Contaminant values were compared to those listed in various literature and agency sources, both regional and national, including the National Contaminant Biomonitoring Program (NCBP), in which the 85th percentile and above represents what is considered to be an elevated contaminant concentration and cause for concern. Generally, bivalves were at the high end of their ranges for both organic and inorganic contaminants. In this study, OCs were detectable in 67 percent of fish from the Mississippi River site, ranging from 0.15 to 1.09 μg/g wet weight (ww) or fresh weight (fw), and in 11 percent of the fish from the marsh sites, ranging from 0.06 to 0.612 μg/g ww. Bivalves from the Mississippi River had OC levels of 0.096 μg/g ww, whereas none were detectable in bivalves at the marsh sites. In this study, p,p ’-dichlorodiphenyldichloroethylene ( p,p’ -DDE) (a biodegradation product of DDT [dichlorodiphenyl trichloroethane]) and total PCBs were the most frequently detected OCs and were primarily from the Mississippi River. For total OC content, using adjusted least squares means, some significant interactions were noted between fish species and sites. PAHs were detected in aquatic animals at all sites (range of 0.017–17.534 μg/g ww), as were AHs (range of 0.423–4.549 μg/g ww); the highest levels of PAHs and AHs were found in bivalves from the Mississippi River. When analysis of variance (α = 0.05) was performed with data from aquatic animals, there were only two significant relationships between PAHs, AHs, and OCs between species, site, and age or the interaction among these variables. There was an interaction between fish species and n-decane (an AH) in that mullet and largemouth bass had significantly higher levels than did catfish ( P = 0.0175). When general linear means were used to investigate associations of inorganic contaminants among fish species, site, and age or any interactions among these variables, no significant results were noted for arsenic, cadmium, lead, beryllium, boron, molybdenum, or nickel. The range of mercury in fish in this study was 0.04–0.14 μg/g ww (0.14– 0.48 μg/g dry weight [dw]), with the most elevated levels detected in predatory largemouth bass at the sampling point farthest downstream from the structure and within the marsh area. Mercury was positively correlated with fish age ( P = 0.0152), where levels were estimated to increase 0.0253 parts per million (ppm) dw per year. In the Mississippi River, catfish showed significantly higher levels of mercury than did mullet or largemouth bass ( P = 0.00167). Among fish species, mullet displayed the highest levels in fish of aluminum, barium, manganese, and iron, all considered to have low toxicity in hydrologic systems. An interaction between fish and site was seen with aluminum ( P = 0.0031), where concentrations in mullet were significantly higher in the Mississippi River than at the other sites, as was also seen with barium ( P = 0.0009), chromium ( P = <0.0001), manganese ( P = 0.0004), strontium ( P = 0.0074), vanadium ( P = 0.0156), and zinc ( P = 0.0059). For iron ( P = 0.0.0001), mullet and largemouth bass at both the Mississippi River and Lake Salvador showed higher levels than did catfish, and these two species showed higher levels at two of the four sites. An interaction between fish and site was also seen with chromium ( P = <0.0001) in that concentrations in mullet were significantly higher in the Mississippi River than at the other sites, as was also seen with strontium ( P = 0.0074), vanadium ( P = 0.0156), and zinc ( P = 0.0059), metals for which deleterious effects have been demonstrated in other ecosystems. The NCBP program lists the 85th percentile for zinc at 34.2 μg/g fw (117.9 μg/g dw). In the Davis Pond prediversion biomonitoring study (hereafter the current study), one fish (MUL31RIVER, fish ID 8) showed values higher than that (125.4 μg/g dw or 37.54 μg/g ww), and the Mississippi River bivalve sample (MUSSRIVER) had a value of 140 μg/g dw (41.2 μg/g ww). In the current study, approximately 86 percent of the fish had measurable selenium levels, yet none reached the 85th percentile. The 85th percentile for selenium from the NCBP was 0.73 μg/g ww. Significantly higher levels of selenium were seen in mullet than in largemouth bass and catfish ( P = 0.0023). The NCBP 85th percentile for lead is 0.22 μg/g ww (0.76 μg/g dw). In the current study, the range of concentrations of lead was as much as 18.3 ppm dw (MUL31RIVER, fish ID 8), with the three most elevated values (range of 3.46–5.31 μg/g ww) coming from mullet from the Mississippi River. Biomarker data are measurable and directly reflect the condition of the animal, and measuring more than one biomarker in an individual increases confidence in health assessments. In the current study, biomarkers included macrophage aggregates (MAs), liver (hepatosomatic index [HSI]) and spleen (splenosomatic index [SSI]) weight to body weight ratios, percent white blood cells (WBCs) in whole blood, and condition factor. Few significant differences were noted with any of the biomarkers between sites, and there were no relationships between species and sites. For improved use of biomarker assessments, an increase in fish sample size would be useful for postdiversion sampling, as would comparisons of fish of the same sex and reproductive condition. During the current study, success for eagle nests in the diversion area and reference sites was similar as determined by numbers of nestlings fledged. When temperatures were below average during winter 2000, nests in both regions similarly failed. At each nest, the primary evidence of food items was small mammals. Eaglets (n = 6) generally appeared healthy, and whole blood concentrations of organic contaminants exceeded detection limits with three incidences of p,p’ -DDE (0.002–0.006 μg/L ww) and one incidence of oxychlordane (0.002 μg/L ww). The levels of p,p’ -DDE were well below those that have been inversely correlated with productivity and success rates of nesting bald eagles on a regional scale. The low values found in the whole blood samples for OC pesticides and PCBs were even lower when corrected for plasma volume. Aluminum values were 3.66 and 5.75 μg/L in two samples, zinc ranged from 5.21 to 6.77 μg/L ww in six samples, and silicon ranged from 1.7 to 4.6 μg/L in four samples. Selenium was detectable in each bird with the range at 0.332–0.566 μg/L ww, and strontium ranged from 0.0581 to 0.0975 μg/L ww. Mercury was detectable in blood samples from each bird and ranged from 0.0254 to 0.0845 μg/L ww, whereas lead was detectable in four samples and ranged from 0.0042 to 0.0136 μg/L ww. Although no detectable levels of total PCBs were found (also correlated with decreased reproductive productivity), 70 percent of the aquatic animals from the Mississippi River contained total PCBs (range 0.13–0.79 μg/L), whereas only about 7 percent of the aquatic animals sampled from the marsh area contained PCBs. Suggestions for postdiversion sampling include lowering the analytical detection limit for some metals, sampling aquatic animals over the course of a single season, obtaining a higher sample number of mature fish of one species (for example, blue catfish) within a range of total lengths for biomarker analyses, obtaining otoliths for estimating fish ages, assessing dioxins in eaglet blood, examining triazines in water, and obtaining all Mississippi River fish samples as close to the Davis Pond structure intake as possible. Because contaminants found in blood of eaglets reflect their prey species and because of the contaminant levels found in fish in the current study, eaglets may not be consuming primarily these species; therefore, obtaining juvenile nutria ( Myocastor coypus ) or turtle species for contaminant analyses might be considered, as well as collecting greater blood volume and using plasma to measure OCs and PCBs. Data obtained postdiversion will be compared with prediversion data to monitor changes.

Louisiana

Ubiquitous tar balls with a California-source signature on the shorelines of Prince William Sound, Alaska

Although the shorelines of Prince William Sound still bear traces of the 1989 Exxon Valdez oil spill, most of the flattened tar balls that can be found today on these shorelines are not residues of Exxon Valdez oil. Instead, the carbon-isotopic and hydrocarbonbiomarker signatures of 61 tar ball samples, collected from shorelines throughout the northern and western parts of the sound, are all remarkably similar and have characteristics consistent with those of oil products that originated from the Monterey Formation source rocks of California. The carbonisotopic compositions of the tar balls are all closely grouped (<513Cpdb = -23.7 ± 0.2%o), within the range found in crude oils from those rocks, but are distinct from isotopic compositions of 28 samples of residues from the Exxon ValdezoW spill (<513Cpdb = -29.4 ± 0.1%o). Likewise, values for selected biomarker ratios in the tar balls are all similar but distinct from values of residues from the 1989 oil spill. Carbon-isotopic and biomarker signatures generally relate the tar balls to oil products used in Alaska before ~1970 for construction and pavements. How these tar balls with such similar geochemical characteristics became so widely dispersed throughout the northern and western parts of the sound is not known with certainty, but the great 1964 Alaska earthquake was undoubtedly an important trigger, causing spills from ruptured storage facilities of California-sourced asphalt and fuel oil into Prince William Sound.

Alaska

Reconnaissance of 17 beta-estradiol, 11-ketotestosterone, vitellogenin, and gonad histopathology in common carp of United States streams; potential for contaminant-induced endocrine disruption

A reconnaissance of sex steroid hormones and other biomarkers in common carp was used to assess whether endocrine disruption may be occurring in fish in United States streams, to evaluate relations between endocrine disruption and contaminant levels, and to determine requirements for further studies. 17?-estradiol, 11-ketotestosterone, vitellogenin, and gonadal histopathology were measured in adult carp (usually 10--15 for each sex) at 25 sites (647 fish), representing a wide range of environmental settings typical of major regions of the nation. Fish were collected during August--December 1994, a period of gonadal maturation after spawning. Contaminants evaluated were organochlorine pesticides and polychlorinated biphenyls in tissue; phthalates, phenols, and polycyclic aromatic hydrocarbons in bed sediment; and dissolved pesticides in water. Mean site concentrations of steroid hormones spanned two orders of magnitude for both sexes. No significant regional differences in steroid hormones were detected for males, but females from the Northern and Southern Midcontinent were significantly different from other regions of the country in one or both hormones. Within all regions there were significant differences between sites in one or both hormones for both sexes. Most correlation coefficients between biomarkers and contaminants were negative. Contaminants that had significant (a=0.05) correlations with biomarkers were organochlorine pesticides, phenols, and dissolved pesticides. The strongest pattern common to both males and females was a negative correlation between the hormone ratio (E2/11-KT) and dissolved pesticides. The significant site-to-site differences in biomarkers, and the presence of significant correlations between biomarkers and contaminants, are evidence that fish in some streams may be experiencing endocrine disruption. Improved information is needed to evaluate whether endocrine disruption is actually occurring and if there are reproductive effects on individual or populations of carp or other species. Future studies should shift to more intensive study of fewer sites, including reference and contaminated sites, in order to address these additional questions.

Open-File Report

Monitoring nearshore ecosystem health using Pacific razor clams (Siliqua patula) as an indicator species

An emerging approach to ecosystem monitoring involves the use of physiological biomarker analyses in combination with gene transcription assays. For the first time, we employed these tools to evaluate the Pacific razor clam ( Siliqua patula ), which is important both economically and ecologically, as a bioindicator species in the northeast Pacific. Our objectives were to (1) develop biomarker and gene transcription assays with which to monitor the health of the Pacific razor clam, (2) acquire baseline biomarker and gene transcription reference ranges for razor clams, (3) assess the relationship between physiological and gene transcription assays and (4) determine if site-level differences were present. Pacific razor clams were collected in July 2015 and 2016 at three sites within each of two national parks in southcentral Alaska. In addition to determining reference ranges, we found differences in biomarker assay and gene transcription results between parks and sites which indicate variation in both large-scale and local environmental conditions. Our intent is to employ these methods to evaluate Pacific razor clams as a bioindicator of nearshore ecosystem health. Links between the results of the biomarker and gene transcription assays were observed that support the applicability of both assays in ecosystem monitoring. However, we recognize the need for controlled studies to examine the range of responses in physiology and gene transcripts to different stressors. sors.

Alaska

Arsenic-related oxidative stress in experimentally dosed wild great tit nestlings

Arsenic (As) is broadly distributed due to natural and anthropogenic sources, and it may cause adverse effects in birds. However, research on other elements (Pb, Hg and Cd) has been prioritized, resulting in scarce data on As exposure and related effects in wild birds. One of the mechanisms responsible for As toxicity is oxidative stress. Therefore, the aim of this study was to investigate if environmentally relevant As levels affected oxidative stress biomarkers in great tits ( Parus major ). This is the first field experiment studying the effects of As on oxidative stress in wild passerines. Wild great tit nestlings were orally dosed with sodium arsenite (Control: water, Low dose: 0.2 μg g −1 d −1 and High dose: 1 μg g −1 d −1 ; from day 3 to day 13 post-hatching). We intended to reach As concentrations similar to those at which passerines are exposed to at actual polluted areas. We compared the responses to the experimental manipulations (High, Low and Control groups) with those in an As/metal-exposed population breeding close to a Cu–Ni smelter in Finland (Smelter group). A set of antioxidants (tGSH, GSH:GSSG ratio, CAT, SOD, GST and GPx), and oxidative damage biomarkers (lipid peroxidation, protein carbonylation, 8-hydroxy-2′-deoxyguanosine formation in DNA, and telomere length) were explored in blood. Arsenic administration had no significant effect on most of the biomarkers measured: only the CAT activity was lower in the High As group and the GPx activity was enhanced in the Smelter group compared to the Control. Our results suggest that the dose and duration of the As exposure was not enough to induce oxidative damage in red cells of great tit nestlings. In spite of this, nestlings dosed with 1 μg g −1 d −1 of sodium arsenite showed non-significantly higher oxidative stress biomarkers than controls, suggesting that we were close to an effect level for the redox-defense system. Oxidative effects at equivalent As levels combined with other stressors cannot be dismissed.

Harjavalta

Transcriptome signatures of wastewater effluent exposure in larval zebrafish vary with seasonal mixture composition in an effluent-dominated stream

Wastewater treatment plant (WWTP) effluent-dominated streams provide critical habitat for aquatic and terrestrial organisms but also continually expose them to complex mixtures of pharmaceuticals that can potentially impair growth, behavior, and reproduction. Currently, few biomarkers are available that relate to pharmaceutical-specific mechanisms of action. In the experiment reported in this paper, zebrafish ( Danio rerio ) embryos at two developmental stages were exposed to water samples from three sampling sites (0.1 km upstream of the outfall, at the effluent outfall, and 0.1 km below the outfall) during base-flow conditions from two months (January and May) of a temperate-region effluent-dominated stream containing a complex mixture of pharmaceuticals and other contaminants of emerging concern. RNA-sequencing identified potential biological impacts and biomarkers of WWTP effluent exposure that extend past traditional markers of endocrine disruption. Transcriptomics revealed changes to a wide range of biological functions and pathways including cardiac, neurological, visual, metabolic, and signaling pathways. These transcriptomic changes varied by developmental stage and displayed sensitivity to variable chemical composition and concentration of effluent, thus indicating a need for stage-specific biomarkers. Some transcripts are known to be associated with genes related to pharmaceuticals that were present in the collected samples. Although traditional biomarkers of endocrine disruption were not enriched in either month, a high estrogenicity signal was detected upstream in May and implicates the presence of unidentified chemical inputs not captured by the targeted chemical analysis. This work reveals associations between bioeffects of exposure, stage of development, and the composition of chemical mixtures in effluent-dominated surface water. The work underscores the importance of measuring effects beyond the endocrine system when assessing the impact of bioactive chemicals in WWTP effluent and identifies a need for non-targeted chemical analysis when bioeffects are not explained by the targeted analysis.

Iowa

Non-lethal assessment of freshwater mussel physiological response to changes in environmental factors

The development of effective nonlethal biomonitoring techniques is imperative for the preservation of imperiled freshwater mussel populations. Changes in hemolymph chemistry profiles and tissue glycogen are potential biomarkers for nonlethally monitoring stress in mussels. We sampled three species in the Flint River Basin over 2 years to evaluate how these hemolymph and tissue biomarkers responded to environmental changes. We used hierarchical linear models to evaluate the relationships between variation in the biomarkers and environmental factors and found that the responses of the hemolymph and tissue parameters were strongly related to stream discharge. Shifts in alanine aminotransferase and glycogen showed the largest relations with discharge at the time of sampling, while magnesium levels were most explained by the discharge for 5 days prior to sampling. Aspartate aminotransferase, bicarbonate, and calcium showed the strongest relations with mean discharge for 15 days prior to sampling. The modeling results indicated that biomarker responses varied substantially among individuals of different size, sex, and species and illustrated the value of hierarchical modeling techniques to account for the inherent complexity of aquatic ecosystems.

Canadian Journal of Fisheries and Aquatic Sciences

Oil and gas geochemistry and petroleum systems of the Fort Worth Basin

Detailed biomarker and light hydrocarbon geochemistry confirm that the marine Mississippian Barnett Shale is the primary source rock for petroleum in the Fort Worth Basin, north-central Texas, although contributions from other sources are possible. Biomarker data indicate that the main oil-generating Barnett Shale facies is marine and was deposited under dysoxic, strong upwelling, normal salinity conditions. The analysis of two outcrop samples and cuttings from seven wells indicates variability in the Barnett Shale organic facies and a possibility of other oil subfamilies being present. Light hydrocarbon analyses reveal significant terrigenous-sourced condensate input to some reservoirs, resulting in terrigenous and mixed marine-terrigenous light hydrocarbon signatures for many oils. The light hydrocarbon data suggest a secondary, condensate-generating source facies containing terrigenous or mixed terrigenous-marine organic matter. This indication of a secondary source rock that is not revealed by biomarker analysis emphasizes the importance of integrating biomarker and light hydrocarbon data to define petroleum source rocks. Gases in the Fort Worth Basin are thermogenic in origin and appear to be cogenerated with oil from the Barnett Shale, although some gas may also originate by oil cracking. Isotope data indicate minor contribution of biogenic gas. Except for reservoirs in the Pennsylvanian Bend Group, which contain gases spanning the complete range of observed maturities, the gases appear to be stratigraphically segregated, younger reservoirs contain less mature gas, and older reservoirs contain more mature gas. We cannot rule out the possibility that other source units within the Fort Worth Basin, such as the Smithwick Shale, are locally important petroleum sources. Copyright ?? 2007. The American Association of Petroleum Geologists. All rights reserved.

American Association of Petroleum Geologists Bulle

Biomonitoring of Environmental Status and Trends (BEST) Program: Environmental contaminants and their effects on fish in the Mississippi River Basin

We collected, examined, and analyzed 1378 fish of 22 species from 47 sites in the Mississippi River basin (MRB) during 1995 and from a reference site in 1996. The sampling sites in the MRB represented National Contaminant Biomonitoring Program (NCBP) stations situated at key points on major rivers and National Water- Quality Assessment Program (NAWQA) stations located on lower-order rivers and streams in the Eastern Iowa Basins (EIB) and Mississippi Embayment (MSE) Study Units. The reference site was the water supply system of the USGS-Leetown Science Center in rural Jefferson County, WV. Common carp (Cyprinus carpio; carp) and black basses (Micropterus spp.; bass), the targeted species, together represented 82% of the fish collected. Each fish was examined in the field for externally and internally visible gross lesions, selected organs were weighed to compute various ponderal and organo-somatic indices, and selected tissues and fluids were obtained and preserved for analysis of biomarkers. Fish health indicators included splenic macrophage aggregates, lysozyme activity, and hispathological analysis of liver, kidney, and other tissues. Reproductive biomarkers included analysis of plasma concentrations of vitellogenin (vtg) and the sex steroid hormones 17-estradiol (E2) and 11-ketotestosterone (11- kt); and the histological determination of percent oocyte atresia (in female fish) and gonadal stage. Hepatic ethoxyresorufin O-deethylase (EROD) activity was also measured. Composite samples of whole fish from each station were grouped by species and gender and analyzed for persistent organochlorine and elemental contaminants and for dioxin-like activity (TCDD-EQ) using the H4IIE rat hepatoma cell bioassay. Organochlorine and inorganic contaminant concentrations in fish were generally low relative to historical levels at most sites, but remained present at concentrations representing threats to piscivorous wildlife in some locations. Toxaphene and DDT (mostly as p,p?-DDE) concentrations remained elevated in fish from the cottongrowing regions of the lower Mississippi valley, and were generally greater in the smaller streams draining agricultural areas (that is, in the MSE Study Unit) than at large river sites. Cyclodiene pesticide concentrations were also greatest in the EIB Study Unit and elsewhere in the corn-growing regions of the mid-MRB. Former point-sources of organochlorine pesticides also remained evident, especially in the Mississippi River near Memphis, TN. Consistent with previous findings, total PCB concentrations tended to be greatest (1-3 g/g) in the industrialized and urbanized Ohio River and Upper Mississippi sub-basins and at Memphis, TN, and were generally correlated with TCDD-EQ and EROD activity. Conversely, PCB concentrations were low (<0.1 g/g) in the more agricultural parts of the MRB. Concentrations of inorganic contaminants were also relatively low and stable or declining relative to past levels at most sites. Exceptions were Hg and Se; Hg concentrations were slightly elevated (>0.3 g/g) in bass from the Mississippi River at Memphis and several other sites and in carp from one MSE site. Concentrations of Se were also great enough to constitute a hazard to piscivorous wildlife (>0.6 g/g) at several MRB sites in the western parts of the MRB and were especially high (4-5 g/g) in fish from John Martin Reservoir, CO, where elevated concentrations were reported previously. Biomarker results indicated that fish from many stations had been exposed to contaminants, but at no sites did findings indicate exposure to high concentrations of toxic chemicals. Noteworthy among biomarker findings was that 73% of the male smallmouth bass (Micropterus dolomieui) from the Mississippi River at Lake City, MN (Lake Pepin) were intersex as indicated by the histological detection of ovotestes; and the combined EROD and H4IIE results indicated that fish from several rural sites in the

Mississippi River basin

Effects of Bromus tectorum invasion on microbial carbon and nitrogen cycling in two adjacent undisturbed arid grassland communities

Soil nitrogen (N) is an important component in maintaining ecosystem stability, and the introduction of non-native plants can alter N cycling by changing litter quality and quantity, nutrient uptake patterns, and soil food webs. Our goal was to determine the effects of Bromus tectorum (C 3 ) invasion on soil microbial N cycling in adjacent non-invaded and invaded C 3 and C 4 native arid grasslands. We monitored resin-extractable N, plant and soil δ 13 C and δ 15 N, gross rates of inorganic N mineralization and consumption, and the quantity and isotopic composition of microbial phospholipid biomarkers. In invaded C 3 communities, labile soil organic N and gross and net rates of soil N transformations increased, indicating an increase in overall microbial N cycling. In invaded C 4 communities labile soil N stayed constant, but gross N flux rates increased. The δ 13 C of phospholipid biomarkers in invaded C 4 communities showed that some portion of the soil bacterial population preferentially decomposed invader C 3 -derived litter over that from the native C 4 species. Invasion in C 4 grasslands also significantly decreased the proportion of fungal to bacterial phospholipid biomarkers. Different processes are occurring in response to B. tectorum invasion in each of these two native grasslands that: 1) alter the size of soil N pools, and/or 2) the activity of the microbial community. Both processes provide mechanisms for altering long-term N dynamics in these ecosystems and highlight how multiple mechanisms can lead to similar effects on ecosystem function, which may be important for the construction of future biogeochemical process models.

Biogeochemistry