Christopher J. Schmitt

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.

The H4IIE Cell Bioassay as an Indicator of Dioxin-like Chemicals in Wildlife and the Environment

ABSTRACT The H4IIE cell bioassay has proven utility as a screening tool for planar halogenated hydrocarbons (PHHs) and structurally similar chemicals accumulated in organisms from the wild. This bioassay has additional applications in hazard assessment of PHH exposed populations. In this review, the toxicological principles, current protocols, performance criteria, and field applications for the assay are described. The H4IIE cell bioassay has several advantages over the analytical measurement of PHHs in environmental samples, but conclusions from studies can be strengthened when both bioassay and analytical chemistry data are presented together. Often, the bioassay results concur with biological effects in organisms and support direct measures of PHHs. For biomonitoring purposes and prioritization of PHH-contaminated environments, the H4IIE bioassay may be faster and less expensive than analytical measurements. The H4IIE cell bioassay can be used in combination with other biomarkers such as in vivo measurements of CYP1A1 induction to help pinpoint the sources and identities of dioxin-like chemicals. The number of studies that measure H4IIE-derived TCDD-EQs continues to increase, resulting in subtle improvements over time. Further experiments are required to determine if TCDD-EQs derived from mammalian cells are adequate predictors of toxicity to non-mammalian species. The H4IIE cell bioassay has been used in over 300 published studies, and its combination of speed, simplicity, and ability to integrate the effects of complex containment mixtures makes it a valuable addition to hazard assessment and biomonitoring studies.

Ecological impacts of lead mining on Ozark streams: toxicity of sediment and pore water.

We studied the toxicity of sediments downstream of lead-zinc mining areas in southeast Missouri, using chronic sediment toxicity tests with the amphipod, Hyalella azteca, and pore-water toxicity tests with the daphnid, Ceriodaphnia dubia. Tests conducted in 2002 documented reduced survival of amphipods in stream sediments collected near mining areas and reduced survival and reproduction of daphnids in most pore waters tested. Additional amphipod tests conducted in 2004 documented significant toxic effects of sediments from three streams downstream of mining areas: Strother Creek, West Fork Black River, and Bee Fork. Greatest toxicity occurred in sediments from a 6-km reach of upper Strother Creek, but significant toxic effects occurred in sediments collected at least 14 km downstream of mining in all three watersheds. Toxic effects were significantly correlated with metal concentrations (nickel, zinc, cadmium, and lead) in sediments and pore waters and were generally consistent with predictions of metal toxicity risks based on sediment quality guidelines, although ammonia and manganese may also have contributed to toxicity at a few sites. Responses of amphipods in sediment toxicity tests were significantly correlated with characteristics of benthic invertebrate communities in study streams. These results indicate that toxicity of metals associated with sediments contributes to adverse ecological effects in streams draining the Viburnum Trend mining district.

Mercury trends in fish from rivers and lakes in the United States, 1969-2005

A national dataset on concentrations of mercury in fish, compiled mainly from state and federal monitoring programs, was used to evaluate trends in mercury (Hg) in fish from US rivers and lakes. Trends were analyzed on data aggregated by site and by state, using samples of the same fish species and tissue type, and using fish of similar lengths. Site-based trends were evaluated from 1969 to 2005, but focused on a subset of the data from 1969 to 1987. Data aggregated by state were used to evaluate trends in fish Hg concentrations from 1988 to 2005. In addition, the most recent Hg fish data (1996–2005) were compared to wet Hg deposition data from the Mercury Deposition Network (MDN) over the same period. Downward trends in Hg concentrations in fish from data collected during 1969–1987 exceeded upward trends by a ratio of 6 to 1. Declining Hg accumulation rates in sediment and peat cores reported by many studies during the 1970s and 1980s correspond with the period when the most downward trends in fish Hg concentrations occurred. Downward Hg trends in both sediment cores and fish were also consistent with the implementation of stricter regulatory controls of direct releases of Hg to the atmosphere and surface waters during the same period. The southeastern USA had more upward Hg trends in fish than other regions for both site and state aggregated data. Upward Hg trends in fish from the southeastern USA were associated with increases in wet deposition in the region and may be attributed to a greater influence of global atmospheric Hg emissions in the southeastern USA. No significant trends were found in 62% of the fish species from six states from 1996 to 2005. A lack of Hg trends in fish in the more recent data was consistent with the lack of trends in wet Hg deposition at MDN sites and with relatively constant global emissions during the same time period. Although few significant trends were observed in the more recent Hg concentrations in fish, it is anticipated that Hg concentrations in fish will respond to changes in atmospheric Hg deposition, however, the magnitude and timing of the response is uncertain.

Environmental contaminants in freshwater fish and their risk to piscivorous wildlife based on a national monitoring program

Organochlorine chemical residues and elemental concentrations were measured in piscivorous and benthivorous fish at 111 sites from large U.S. river basins. Potential contaminant sources such as urban and agricultural runoff, industrial discharges, mine drainage, and irrigation varied among the sampling sites. Our objectives were to provide summary statistics for chemical contaminants and to determine if contaminant concentrations in the fish were a risk to wildlife that forage at these sites. Concentrations of dieldrin, total DDT, total PCBs, toxaphene, TCDD-EQ, cadmium, chromium, mercury, lead, selenium, and zinc exceeded toxicity thresholds to protect fish and piscivorous wildlife in samples from at least one site; most exceedences were for total PCBs, mercury, and zinc. Chemical concentrations in fish from the Mississippi River Basin exceeded the greatest number of toxicity thresholds. Screening level wildlife risk analysis models were developed for bald eagle and mink using no adverse effect levels (NOAELs), which were derived from adult dietary exposure or tissue concentration studies and based primarily on reproductive endpoints. No effect hazard concentrations (NEHC) were calculated by comparing the NOAEL to the food ingestion rate (dietary-based NOAEL) or biomagnification factor (tissue-based NOAEL) of each receptor. Piscivorous wildlife may be at risk from a contaminant if the measured concentration in fish exceeds the NEHC. Concentrations of most organochlorine residues and elemental contaminants represented no to low risk to bald eagle and mink at most sites. The risk associated with pentachloroanisole, aldrin, Dacthal, methoxychlor, mirex, and toxaphene was unknown because NOAELs for these contaminants were not available for bald eagle or mink. Risk differed among modeled species and sites. Our screening level analysis indicates that the greatest risk to piscivorous wildlife was from total DDT, total PCBs, TCDD-EQ, mercury, and selenium. Bald eagles were at greater risk to total DDT and total PCBs than mink, whereas risks of TCDD-EQ, mercury, and selenium were greater to mink than bald eagle.

Natural and Anthropogenic Influences on the Distribution of the Threatened Neosho Madtom in a Midwestern Warmwater Stream

Abstract We attempted to discern the contributions of physical habitat, water chemistry, nutrients, and contaminants from historic lead–zinc mining activities on the riffle-dwelling benthic fish community of the Spring River, a midwestern warmwater stream that originates in Missouri and flows into Kansas and Oklahoma. The Spring River has a fish community that includes the Neosho madtom Noturus placidus, a species federally listed as threatened. Although anthropogenic factors such as contaminants limited populations and densities of fishes, an integrated assessment of natural and anthropogenic factors was necessary to effectively estimate the influence of the latter. Fish populations in the Spring River, especially Neosho madtoms, seem to be limited by the presence of cadmium, lead, and zinc in water and in benthic invertebrate food sources and by physical habitat. The population density and community structure of fish in the Spring River also seem to be related to water chemistry and nutrients. Concurren...

INHIBITION OF ERYTHROCYTE δ-AMINOLEVULINIC ACID DEHYDRATASE (ALAD) ACTIVITY IN FISH FROM WATERS AFFECTED BY LEAD SMELTERS

We assessed the effects on fish of lead (Pb) released to streamsby smelters located in Trail, BC (Canada), E. Helena, MT, Herculaneum, MO, and Glover, MO. Fish were collected by electrofishing from sites located downstream of smelters and from reference sites. Blood from each fish was analyzed for δ-aminolevulinic acid dehydratase (ALAD) activity and hemoglobin (Hb), and samples of blood, liver, or carcass were analyzed for Pb, zinc (Zn), or both. Fish collected downstreamof all four smelters sites had elevated Pb concentrations, decreased ALAD activity, or both relative to their respectivereference sites. At E. Helena, fish from the downstream site also had lower Hb concentrations than fish from upstream. Differences among taxa were also apparent. Consistent with previous studies, ALAD activity in catostomids (Pisces: Catostomidae-northern hog sucker, Hypentelium nigricans;river carpsucker, Carpiodes carpio; largescale sucker, Catostomus macrocheilus; and mountain sucker, C. platyrhynchus) seemed more sensitive to Pb-induced ALAD inhibition than the salmonids (Pisces: Salmonidae-rainbow trout,Oncorhynchus mykiss; brook trout, Salvelinus fontinalis) or common carp (Cyprinus carpio). Some of these differences may have resulted from differential accumulation of Zn, which was not measured at all sites. We detected no ALAD activity in channel catfish (Ictaluruspunctatus) from either site on the Mississippi River at Herculaneum, MO. Our findings confirmed that Pb is releasedto aquatic ecosystems by smelters and accumulated by fish, andwe documented potentially adverse effects of Pb in fish. We recommend that Zn be measured along with Pb when ALAD activityis used as a biomarker and the collection of at least 10 fish ofa species at each site to facilitate statistical analysis.

Comparison of an enzyme-linked immunosorbent assay (ELISA) to gas chromatography (GC)--measurement of polychlorinated biphenyls (PCBs) in selected US fish extracts.

The analysis of PCBs in fish tissues by immunoassay methods was evaluated using fish collected from a US monitoring program, the National Contaminant Biomonitoring Program of the US Department of Interior, Fish and Wildlife Service. Selected composite whole fish samples, which represented widely varying concentrations and sources of PCBs, were extracted and subjected to congener PCB analysis by gas chromatography (GC) and total PCB analysis using an ELISA (ePCBs) calibrated against technical Aroclor 1248. PCB congener patterns in these fishes were different from the patterns found in commercial Aroclors or their combinations as demonstrated by principal component analysis of normalized GC congener data. The sum of the PCB congeners measured by GC (total-PCBs) ranged from 37 to 4600 ng/g (wet weight). Concentrations of PCBs as determined by the ELISA method were positively correlated with total-PCBs and the ePCBs/total-PCBs ratios for individual samples ranged from 1 to 6. Ratios of ePCBs/total-PCBs for dilutions of Aroclors 1242, 1254, and 1260 and for matrix spikes range from 0.6 for 1242 to 2.5 for 1254 and 1260. These results suggest that higher chlorinated PCB congeners have higher affinity for the anti-PCB antibodies. Partial least squares with latent variable analysis of GC and ELISA data of selected Aroclors and fish samples also support the conclusion that ELISA derived PCB concentrations are dependent on the degree of chlorination.

A LONGITUDINAL ASSESSMENT OF THE AQUATIC MACROINVERTEBRATE COMMUNITY IN THE CHANNELIZED LOWER MISSOURI RIVER

We conducted an aquatic macroinvertebrate assessment in the channelized reach of the lower Missouri River, and used statistical analysis of individual metrics and multimetric scores to identify community response patterns and evaluate relative biological condition. We examined longitudinal site differences that are potentially associated with water qualityrelated factors originating from the Kansas City metropolitan area, using data from coarse rock substrate in flowing water habitats (outside river bends), and depositional mud substratein slack water habitats (dike fields). Three sites above rivermile (RM) 369 in Kansas City (Nebraska City, RM = 560; St. Joseph, RM = 530; Parkville, RM = 377) and three below (Lexington, RM = 319; Glasgow, RM = 228; Hermann, RM = 94) were sampled with rock basket artificial substrates, a qualitative kicknet method, and the Petite Ponar. We also compared the performance of the methods used. A total of 132 aquatic macroinvertebrate taxa were collected from the lower Missouri River; one third of these taxa belonged to the sensitiveEPOT insect orders (Ephemeroptera, Plecoptera, Odonata, and Trichoptera). Rock baskets had the highest mean efficiency (34.1%) of the methods, and the largest number of taxa was collected by Ponar (n = 69) and kicknet (n = 69) methods. Seven of the 15 metrics calculated from rock basket data, and five ofthe nine metrics calculated from Ponar data showed highly significant differences (ANOVA, P < 0.001) at one or more sitesbelow Kansas City. We observed a substantial reduction in net-spinning Trichoptera in rock habitats below Kansas City (Lexington), an increase in relative dominance of Oligochaeta in depositional habitats at the next site downstream (Glasgow), and lower relative condition scores in rock habitat at Lexingtonand depositional habitat at Glasgow. Collectively, these data indicate that some urban-related impacts on the aquatic macroinvertebrate community are occurring. Our results suggest that the methods and assessment framework we used in this studycould be successfully applied on a larger scale with concurrentwater and sediment chemistry to validate metrics, establish impairment levels, and develop a specific macroinvertebrate community index for the lower Missouri River. We recommend accomplishing this with longitudinal multi-habitat sampling at a larger number of sites related to all potential sources of impairment, including major tributaries, urban areas, and point sources.

Biochemical effects of lead, zinc, and cadmium from mining on fish in the Tri-States District of northeastern Oklahoma, USA.

We assessed the exposure of fish from the Spring and Neosho Rivers in northeast Oklahoma, USA, to lead, zinc, and cadmium from historical mining in the Tri-States Mining District (TSMD). Fish (n = 74) representing six species were collected in October 2001 from six sites on the Spring and Neosho Rivers influenced to differing degrees by mining. Additional samples were obtained from the Big River, a heavily contaminated stream in eastern Missouri, USA, and from reference sites. Blood from each fish was analyzed for Pb, Zn, Cd, Fe, and hemoglobin (Hb). Blood also was analyzed for delta-aminolevulinic acid dehydratase (ALA-D) activity. The activity of ALA-D, an enzyme involved in heme synthesis, is inhibited by Pb. Concentrations of Fe and Hb were highly correlated (r = 0.89, p < 0.01) across all species and locations and typically were greater in common carp (Cyprinus carpio) than in other taxa. Concentrations of Pb, Zn, and Cd typically were greatest in fish from sites most heavily affected by mining and lowest in reference samples. The activity of ALA-D, but not concentrations of Hb or Fe, also differed significantly (p < 0.01) among sites and species. Enzyme activity was lowest in fish from mining-contaminated sites and greatest in reference fish, and was correlated negatively with Pb in most species. Statistically significant (p < 0.01) linear regression models that included negative terms for blood Pb explained as much as 68% of the total variation in ALA-D activity, but differences among taxa were highly evident. Positive correlations with Zn were documented in the combined data for channel catfish (Icralurus punctatus) and flathead catfish (Pylodictis olivaris), as has been reported for other taxa, but not in bass (Micropterus spp.) or carp. In channel catfish, ALA-D activity appeared to be more sensitive to blood Pb than in the other species investigated (i.e., threshold concentrations for inhibition were lower). Such among-species differences are consistent with previous studies. Enzyme activity was inhibited by more than 50% relative to reference sites in channel catfish from several TSMD sites. Collectively, our results indicate that Pb is both bioavailable and active biochemically in the Spring-Neosho River system.