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Biomarker Benchmarks: Reproductive and Endocrine Biomarkers in Largemouth Bass and Common Carp from United States Waters

The U.S. Geological Survey (USGS) has developed a national database and report on endocrine and reproductive condition in two species of fish collected in U.S. streams and rivers. This information provides scientists with a national basis for comparing results of endocrine measurements in fish from individual sites throughout the country, so that scientists can better ascertain normal levels of biomarkers. The database includes information on several measures of reproductive and endocrine condition for common carp and largemouth bass. Data summaries are provided by reproductive season and geographic region. A national-scale reconnaissance investigation was initiated in 1994 by the USGS that utilized a suite of biological assays (biomarkers) as indicators of reproductive health, and potentially, endocrine disruption in two widely distributed species of teleost (bony) fish, largemouth bass (Micropterus salmoides) and common carp (Cyrinus carpio). The suite of assays included plasma sex-steroid hormones, stage of gonadal development, and plasma vitellogenin, an egg protein that indicates exposure to estrogenic compounds when found in male fish. More than 2,200 common carp and 650 largemouth bass were collected at 119 rivers and streams (fig. 1).

Fact Sheet

Biomarker generation from Type II-S kerogens in claystone and limestone during hydrous and anhydrous pyrolysis

A claystone and a limestone containing immature Type II-S kerogen were thermally matured in the presence and absence of water, to study the influence of water and clay minerals on the generation of biomarkers. In contrast to hydrous pyrolysis, anhydrous pyrolysis of the claystone did not generate biomarkers, which resulted in the loss of important information. Desulfurization of the polar fraction of the claystone showed that anhydrous pyrolysis is not capable of converting S-bound biomarkers to free biomarkers. For the limestone, the differences between hydrous and anhydrous pyrolysis are less dramatic. Adsorption of the polar fraction of the claystone to smectite interlayers probably leads to cross-linking reactions, preventing the generation of free biomarkers. During hydrous pyrolysis, the smectite interlayers are occupied by water so that generation of biomarkers can take place. In addition, cross-linking reactions during anhydrous pyrolysis of the claystone may be enhanced because of the presence of S-S bonds in the organic matter of the claystone. These results show that water is important in closed system laboratory experiments designed to simulate natural maturation of sedimentary organic matter.A claystone and a limestone containing immature Type II-S kerogen were thermally matured in the presence and absence of water, to study the influence of water and clay minerals on the generation of biomarkers. In contrast to hydrous pyrolysis, anhydrous pyrolysis of the claystone did not generate biomarkers, which resulted in the loss of important information. Desulfurization of the polar fraction of the claystone showed that anhydrous pyrolysis is not capable of converting S-bound biomarkers to free biomarkers. For the limestone, the differences between hydrous and anhydrous pyrolysis are less dramatic. Adsorption of the polar fraction of the claystone to smectite interlayers probably leads to cross-linking reactions, preventing the generation of free biomarkers. During hydrous pyrolysis, the smectite interlayers are occupied by water so that generation of biomarkers can take place. In addition, crosslinking reactions during anhydrous pyrolysis of the claystone may be enhanced because of the presence of S-S bonds in the organic matter of the claystone. These results show that water is important in closed system laboratory experiments designed to simulate natural maturation of sedimentary organic matter.

Conference Paper

Fluid inclusions and biomarkers in the Upper Mississippi Valley zinc-lead district; implications for the fluid-flow and thermal history of the Illinois Basin

The Upper Mississippi Valley zinc-lead district is hosted by Ordovician carbonate rocks at the northern margin of the Illinois Basin. Fluid inclusion temperature measurements on Early Permian sphalerite ore from the district are predominantly between 90?C and I50?C. These temperatures are greater than can be explained by their reconstructed burial depth, which was a maximum of approximately 1 km at the time of mineralization. In contrast to the temperatures of mineral formation derived from fluid inclusions, biomarker maturities in the Upper Mississippi Valley district give an estimate of total thermal exposure integrated over time. Temperatures from fluid inclusions trapped during ore genesis with biomarker maturities were combined to construct an estimate of the district's overall thermal history and, by inference, the late Paleozoic thermal and hydrologic history of the Illinois Basin. Circulation of groundwater through regional aquifers, given sufficient flow rates, can redistribute heat from deep in a sedimentary basin to its shallower margins. Evidence for regional-scale circulation of fluids is provided by paleomagnetic studies, regionally correlated zoned dolomite, fluid inclusions, and thermal maturity of organic matter. Evidence for igneous acti vity contemporaneous with mineralization in the vicinity of the Upper Mississippi Valley district is absent. Regional fluid and heat circulation is the most likely explanation for the elevated fluid inclusion temperatures (relative to maximum estimated burial depth) in the Upper Mississippi Valley district. One plausible driving mechanism and flow path for the ore-forming fluids is groundwater recharge in the late Paleozoic Appalachian-Ouachita mountain belt and northward flow through the Reelfoot rift and the proto- Illinois Basin to the Upper Mississippi Valley district. Warm fluid flowing laterally through Cambrian and Ordovician aquifers would then move vertically upward through the fractures that control sphalerite mineralization in the Upper Mississippi Valley district. Biomarker reactant-product measurements on rock extracts from the Upper Mississippi Valley district define a relatively low level ofthermal maturity for the district, 0.353 for sterane and 0.577 for hopane. Recently published kinetic constants permit a time-temperature relationship to be determined from these biomarker maturities. Numerical calculations were made to simulate fluid heat flow through the fracture-controlled ore zones of the Thompson-Temperly mine and heat transfer to the adjacent rocks where biomarker samples were collected. Calculations that combine the fluid inclusion temperatures and the biomarker constraints on thermal maturity indicate that the time interval during which mineralizing fluids circulated through the Upper Mississippi Valley district is on the order of 200,000 years. Fluid inclusion measurements and thermal maturities from biomarkers in the district reflect the duration of peak temperatures resulting from regional fluid circulation. On the basis of thermal considerations, the timing of fluorite mineralization in southern Illinois, and the northward-decreasing pattern of fluorine enrichment in sediments, we hypothesize that the principal flow direction was northward through the Cambrian and Ordovician aquifers of the Illinois Basin. A basin-scale flow system would result in mass transport (hydrocarbon migration, transport of metals in solution) and energy (heat) transport, which would in turn drive chemical reactions (for example, maturation of organic matter, mineralization, diagenetic reactions) within the Illinois Basin and at its margins.

Bulletin

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

The roles of diet and habitat use in pesticide bioaccumulation by juvenile Chinook Salmon: Insights from stable isotopes and fatty acid biomarkers

Stable isotopes (SI) and fatty acid (FA) biomarkers can provide insights regarding trophic pathways and habitats associated with contaminant bioaccumulation. We assessed relationships between SI and FA biomarkers and published data on concentrations of two pesticides [dichlorodiphenyltrichloroethane and degradation products (DDX) and bifenthrin] in juvenile Chinook Salmon ( Oncorhynchus tshawytscha ) from the Sacramento River and Yolo Bypass floodplain in Northern California near Sacramento. We also conducted SI and FA analyses of zooplankton and macroinvertebrates to determine whether particular trophic pathways and habitats were associated with elevated pesticide concentrations in fish. Relationships between DDX and both sulfur (δ 34 S) and carbon (δ 13 C) SI ratios in salmon indicated that diet is a major exposure route for DDX, particularly for individuals with a benthic detrital energy base. Greater use of a benthic detrital energy base likely accounted for the higher frequency of salmon with DDX concentrations > 60 ng/g dw in the Yolo Bypass compared to the Sacramento River. Chironomid larvae and zooplankton were implicated as prey items likely responsible for trophic transfer of DDX to salmon. Sulfur SI ratios enabled identification of hatchery-origin fish that had likely spent insufficient time in the wild to substantially bioaccumulate DDX. Bifenthrin concentration was unrelated to SI or FA biomarkers in salmon, potentially due to aqueous uptake, biotransformation and elimination of the pesticide, or indistinct biomarker compositions among invertebrates with low and high bifenthrin concentrations. One FA [docosahexaenoic acid (DHA)] and DDX were negatively correlated in salmon, potentially due to a greater uptake of DDX from invertebrates with low DHA or effects of DDX on FA metabolism. Trophic biomarkers may be useful indicators of DDX accumulation and effects in juvenile Chinook Salmon in the Sacramento River Delta.

Archives of Environmental Contamination and Toxico

Environmental contaminants and biomarker responses in fish from the Rio Grande and its U.S. tributaries: Spatial and temporal trends

We collected, examined, and analyzed 368 fish of seven species from 10 sites on rivers of the Rio Grande Basin (RGB) during late 1997 and early 1998 to document temporal and geographic trends in the concentrations of accumulative contaminants and to assess contaminant effects on the fish. Sites were located on the mainstem of the Rio Grande and on the Arroyo Colorado and Pecos River in Texas (TX), New Mexico (NM), and Colorado. Common carp (Cyprinus carpio) and largemouth bass (Micropterus salmoides) were the targeted species. Fish were examined in the field for internal and external visible gross lesions, selected organs were weighed to compute ponderal and organosomatic indices, and samples of tissues and fluids were obtained and preserved for analysis of fish health and reproductive biomarkers. Whole fish from each station were composited by species and gender and analyzed for organochlorine chemical residues and elemental contaminants using instrumental methods, and for 2,3,7,8-tetrachloro dibenzo-p-dioxin-like activity (TCDD-EQ) using the H4IIE rat hepatoma cell bioassay. Overall, fish from lower RGB stations contained greater concentrations of organochlorine pesticide residues and appeared to be less healthy than those from sites in the central and upper parts of the basin, as indicated by a general gradient of residue concentrations and biomarker responses. A minimal number of altered biomarkers and few or no elevated contaminant concentrations were noted in fish from the upper RGB. The exception was elevated concentrations [up to 0.46 ??g/g wet-weight (ww)] of total mercury (Hg) in predatory species from the Rio Grande at Elephant Butte Reservoir, NM, a condition documented in previous studies. Arsenic (As) and selenium (Se) concentrations were greatest in fish from sites in the central RGB; Se concentrations in fish from the Pecos River at Red Bluff Lake, TX and from the Rio Grande at Langtry, TX and Amistad International Reservoir, TX exceeded published fish and wildlife toxicity thresholds. In the lower RGB, residues of p,p???-DDT metabolites (???1.69 ??g/g ww), chlordane-related compounds (???0.21 ??g/g ww), dieldrin (???0.0.05 ??g/g ww), and toxaphene (???2.4 ??g/g ww) were detected in fish from most sites; maximum concentrations were in channel catfish (Ictalurus punctatus) from the Arroyo Colorado at Harlingen, TX. Concentrations of one or more residues exceeded toxicity thresholds for fish and wildlife in fish from this site and from the Rio Grande at Mission, TX and Brownsville, TX; however, concentrations were lower than those reported by previous studies. In addition, the proportional concentrations of p,p???-DDT at all sites were low, indicating weathered DDT rather than the influx of new material. Concentrations of total PCBs (<0.05 ??g/g ww) and TCDD-EQ (???6 pg/g ww) were comparatively low in all samples. Hepatic ethoxyresorufin O-deethylase (EROD) activity in some fish was elevated relative to reference rates at most sites, but was generally lower than previously reported activity in fish from heavily contaminated locations. The comparatively low PCB and TCDD-EQ concentrations together with elevated EROD activity may reflect exposure to polycyclic aromatic hydrocarbons. Reproductive biomarkers were consistent with chronic contaminant exposure at lower RGB sites; comparatively large percentages of intersex male largemouth bass, relatively low gonadosomatic indices, and elevated plasma vitellogenin concentrations in male fish were noted at three of the four stations. Large percentages of atretic eggs were also observed in the ovaries of female common carp from the Rio Grande at Brownsville, TX. Although many of the conditions noted may have other causes in addition to contaminant exposure, the biomarker results for the lower RGB sites are consistent with subtle responses of fish to contaminants, an interpretation supported by the chemical data of this and other investigations.

Science of the Total Environment

A record of hydrocarbon input to San Francisco Bay as traced by biomarker profiles in surface sediment and sediment cores

San Francisco Bay is one of the world's largest urbanized estuarine systems. Its water and sediment receive organic input from a wide variety of sources; much of this organic material is anthropogenically derived. To document the spatial and historical record of the organic contaminant input, surficial sediment from 17 sites throughout San Francisco Bay and sediment cores from two locations Richardson Bay and San Pablo Bay were analyzed for biomarker constituents. Biomarkers, that is, 'molecular fossils', primarily hopanes, steranes, and n-alkanes, provide information on anthropogenic contamination, especially that related to petrogenic sources, as well as on recent input of biogenic material. The biomarker parameters from the surficial sediment and the upper horizons of the cores show a dominance of anthropogenic input, whereas the biomarker profiles at the lower horizons of the cores indicate primarily biogenic input. In the Richardson Bay core the gradual upcore transition from lower maturity background organics to a dominance of anthropogenic contamination occurred about 70-100 years ago and corresponds to the industrial development of the San Francisco Bay area. In San Pablo Bay, the transition was very abrupt, reflecting the complex depositional history of the area. This sharp transition, perhaps indicating a depositional hiatus or erosional period, dated at pre-1952, is clearly visible. Below, the hiatus the biomarker parameters are immature; above, they are mature and show an anthropogenic overlay. Higher concentrations of terrigenous n-alkanes in the upper horizons in this core are indicative of an increase in terrigenous organic matter input in San Pablo Bay, possibly a result of water diversion projects and changes in the fresh water flow into the Bay from the Delta. Alternatively, it could reflect a dilution of organic material in the lower core sections with hydraulic mining debris.

California

Relations between and among contaminant concentrations and biomarkers in black bass (Micropterus spp.) and common carp (Cyprinus carpio) from large U.S. rivers, 1995-2004

Environmental contaminant and biomarker monitoring data from major U.S. river basins were summarized for black bass (Micropterus spp.) and common carp (Cyprinus carpio) sampled over a nine year period. Cumulative frequency distributions revealed taxon differences for many organochlorine residue concentrations, elemental contaminant concentrations, and biomarkers, but few gender differences were evident for chemical concentrations. Concentrations of dacthal, pentachloroanisole, p,p???-DDE, endosulfan sulfate, barium, cadmium, copper, manganese, lead, selenium, vanadium, and zinc were greater in carp than bass, but concentrations of mercury and magnesium were greater in bass. Gender differences were evident in bass for mercury and in carp for zinc, but the differences were small compared to taxon differences. Greater vitellogenin concentrations, 17??-estradiol concentrations, 17??-estradiol/11-ketotestosterone ratios, and percent oocyte atresia in female carp compared to female bass may be related to the sequential spawning of carp. Regression analyses indicated that as much as 78% of biomarker variation was explained by chemical contaminant concentrations. Sites grouped consistently by river basin in the chemical contaminant principal components analysis (PCA) models and were driven by mercury, magnesium, barium, mirex, and oxychlordane. PCA models for the biomarkers did not group the sites by basin for either bass or carp. Statistical analyses and data interpretation were limited by the study design. The implications of these limitations are discussed. Recommendations to be considered during the planning of future monitoring studies include the exclusion of gender- and species-specific sampling for certain chemical contaminants considering analytical methods with appropriate sensitivities; and allowing for the addition of new chemical and biological variables as methods and information needs evolve. ?? The Royal Society of Chemistry.

Journal of Environmental Monitoring

Ethoxyresorufin-O-deethylase (EROD) activity in fish as a biomarker of chemical exposure

This review compiles and evaluates existing scientific information on the use, limitations, and procedural considerations for EROD activity (a catalytic measurement of cytochrome P4501A induction) as a biomarker in fish. A multitude of chemicals induce EROD activity in a variety of fish species, the most potent inducers being structural analogs of 2,3,7,8-tetracholordibenzo- p -dioxin. Although certain chemicals may inhibit EROD induction/activity, this interference is generally not a drawback to the use of EROD induction as a biomarker. The various methods of EROD analysis currently in use yield comparable results, particularly when data are expressed as relative rates of EROD activity. EROD induction in fish is well characterized, the most important modifying factors being fish species, reproductive status and age, all of which can be controlled through proper study design. Good candidate species for biomonitoring should have a wide range between basal and induced EROD activity (e.g., common carp, channel catfish, and mummichog). EROD activity has proven value as a biomarker in a number of field investigations of bleached kraft mill and industrial effluents, contaminated sediments, and chemical spills. Research on mechanisms of CYP1A-induced toxicity suggests that EROD activity may not only indicate chemical exposure, but also may also precede effects at various levels of biological organization. A current research need is the development of chemical exposure-response relationships for EROD activity in fish. In addition, routine reporting in the literature of EROD activity in standard positive and negative control material will enhance confidence in comparing results from different studies using this biomarker.

Critical Reviews in Toxicology

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

Reproductive and Endocrine Biomarkers in Largemouth Bass (Micropterus salmoides) and Common Carp (Cyprinus carpio) from United States Waters

A nationwide reconnaissance investigation was initiated in 1994 to develop and evaluate a suite of reproductive and endocrine biomarkers for their potential to assess reproductive health and status in teleost (bony) fish. Fish collections were made at 119 sites, representing many regions of the country and land- and water-use settings. Collectively, this report will provide a national and regional benchmark and a basis for evaluating biomarkers of endocrine and reproductive function. Approximately 2,200 common carp (Cyprinus carpio) and 650 largemouth bass (Micropterus salmoides) were collected from 1994 through 1997. The suite of biomarkers used for these studies included: the plasma sex-steroid hormones, 17?-estradiol (E2) and 11 ketotestosterone (11KT); the ratio of E2 to 11KT (E2:11KT); plasma vitellogenin (VTG); and stage of gonadal development. This data report provides fish size, stage and reproductive biomarker data for individual fish and for site and regional summaries of these variables.

Data Series

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

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

Fire, vegetation, and Holocene climate in a southeastern Tibetan lake: a multi-biomarker reconstruction from Paru Co

The fire history of the Tibetan Plateau over centennial to millennial timescales is not well known. Recent ice core studies reconstruct fire history over the past few decades but do not extend through the Holocene. Lacustrine sedimentary cores, however, can provide continuous records of local environmental change on millennial scales during the Holocene through the accumulation and preservation of specific organic molecular biomarkers. To reconstruct Holocene fire events and vegetation changes occurring on the southeastern Tibetan Plateau and the surrounding areas, we used a multi-proxy approach, investigating multiple biomarkers preserved in core sediment samples retrieved from Paru Co, a small lake located in the Nyainqentanglha Mountains (29°47′45.6′′ N, 92°21′07.2′′ E; 4845 m a.s.l.). Biomarkers include n -alkanes as indicators of vegetation, polycyclic aromatic hydrocarbons (PAHs) as combustion proxies, fecal sterols and stanols (FeSts) as indicators of the presence of humans or grazing animals, and finally monosaccharide anhydrides (MAs) as specific markers of vegetation burning processes. Insolation changes and the associated influence on the Indian summer monsoon (ISM) affect the vegetation distribution and fire types recorded in Paru Co throughout the Holocene. The early Holocene (10.7–7.5 cal kyr BP) n -alkane ratios demonstrate oscillations between grass and conifer communities, resulting in respective smouldering fires represented by levoglucosan peaks, and high-temperature fires represented by high-molecular-weight PAHs. Forest cover increases with a strengthened ISM, where coincident high levoglucosan to mannosan (L ∕ M) ratios are consistent with conifer burning. The decrease in the ISM at 4.2 cal kyr BP corresponds with the expansion of regional civilizations, although the lack of human FeSts above the method detection limits excludes local anthropogenic influence on fire and vegetation changes. The late Holocene is characterized by a relatively shallow lake surrounded by grassland, where all biomarkers other than PAHs display only minor variations. The sum of PAHs steadily increases throughout the late Holocene, suggesting a net increase in local to regional combustion that is separate from vegetation and climate change.

Climate of the Past

Exposure to and biomarker responses from legacy and emerging contaminants along three drainages in the Milwaukee Estuary, Wisconsin, USA

Legacy contaminants and contaminants of emerging concern (CECs) were assessed in tree swallow ( Tachycineta bicolor ) tissue and diet samples from three drainages in the Milwaukee estuary, Wisconsin, USA, to understand exposures and possible biomarker responses. Two remote Wisconsin lakes were assessed for comparative purposes. Bioaccumulative classes of contaminants, such as polybrominated diphenyl ethers and per- and polyfluoroalkyl substances, while at higher concentrations than the reference lakes, did not vary significantly among sites or among the three drainages. Polycyclic aromatic hydrocarbons were assessed in diet and sediment and were from primarily pyrogenic sources. Ten biomarkers were assessed relative to contaminant exposure. Polychlorinated biphenyls (PCBs) were elevated above reference conditions at all Milwaukee sites but did not correlate with any measured biomarker responses. Only one site, Cedarburg, just downstream from a Superfund site, had elevated PCBs compared to other sites in the Milwaukee estuary. Few non-organochlorine insecticides or herbicides were detected in tree swallow liver tissue, except for the atrazine metabolite desethylatrazine. Few pharmaceuticals and personal care products were detected in liver tissue except for N,N -diethyl-meta-toluamide, iopamidol, and two antibiotics. The present study is one of the most comprehensive assessments to date, along with the previously published Maumee River data, on the exposure and effects of a wide variety of CECs in birds. Environ Toxicol Chem 2024;00:1–22. © 2024 SETAC. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

Wisconsin

Multivariate classification of the crude oil petroleum systems in southeast Texas, USA, using conventional and compositional data analysis of biomarkers

Chemically, petroleum is an extraordinarily complex mixture of different types of hydrocarbons that are now possible to isolate and identify because of advances in geochemistry. Here, we use biomarkers and carbon isotopes to establish genetic differences and similarities among oil samples. Conventional approaches for evaluating biomarker and carbon isotope relative abundances include statistical techniques such as principal component and cluster analysis. Considering that proportions of the different hydrocarbon molecules are relative parts of a laboratory sample, the data are compositional in nature, thus requiring the use of log-ratio approaches for adequate mathematical modeling. We apply both traditional and compositional modeling approaches to crude oil samples from an onshore area of about 50,000 square miles in southeast Texas. The data comprise 177 crude oil samples from producing oil fields that include key biomarkers, elemental, and isotopic values commonly used in source rock correlation studies. Our results indicate that compositional modeling has higher discriminating power and lower uncertainty than the traditional approach, allowing the identification of up to 16 clusters. Each cluster represents one oil family from a source rock organofacies ranging from Carboniferous to Paleogene. The families provide new insights into important petroleum systems in the Texas onshore region of the Gulf of Mexico sedimentary basin.

Texas

New aromatic biomarkers and possible maturity indicators found in New Albany Shale extracts

Aromatic hydrocarbons from benzene extracts of New Albany Shale were characterized. A biomarker that has a molecular weight of 546 and a structural configuration consistent with that of an alkyl-aromatic hydrocarbon (C 40 H 66 ) was tentatively identified. It was found that the relative concentrations of the biomarker are indicative of differing levels of thermal maturity of the shale organic matter. A 40-carbon bicyclic carotenoid (C 40 H 48 ) is proposed as the geochemical precursor of this biomarker. Thermal maturity of the shale organic matter can also be differentiated by observing differences in “fingerprints” as obtained by field-ionization mass spectrometry on the aromatic hydrocarbon fraction. Using this technique, we found that the more mature shale samples from southeastern Illinois contain more low molecular weight extractable aromatic hydrocarbons and the less mature shale samples from northwestern Illinois contain more high molecular weight extractable aromatic hydrocarbons. It was demonstrated that field-ionization and tandem mass spectrometric techniques through fingerprint and individual compound identification, are useful for shale aromatic hydrocarbon fraction characterization and for thermal maturation interpretation.

Illinois

Biomarker similarities between the saline lacustrine Eocene Green River and the Paleoproterozoic Barney Creek Formations

The Paleoproterozoic Barney Creek Formation, which is currently interpreted as a restricted, deep marine paleoenvironment, plays a disproportionate role in our understanding of Proterozoic ocean chemistry and the rise of complex life. The Barney Creek Formation hosts several unusual biomarker features, specifically its methylhopane and carotenoid signatures. Herein, we demonstrate that the saline lacustrine Eocene Green River Formation shares a similar distribution of methylhopanes and carotenoids, which is characteristic of saline lacustrine organic matter more generally. These distinct methylhopane and carotenoid patterns are not observed together in marine organic matter of any geologic age. These results imply a saline lacustrine depositional environment for the Barney Creek Formation, which agrees with earlier but now abandoned depositional models of this formation. As a result, models of Proterozoic ocean chemistry and emergence of complex life that rely on a marine Barney Creek Formation should be re-examined. Alternatively, if Paleoproterozoic marine biomarker signatures resemble those of younger saline lacustrine systems, then this must be recognized to accurately interpret geologic biomarker and paleoenvironmental records.

Colorado, Utah