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Integrated assessment of wastewater treatment plant effluent estrogenicity in the Upper Murray River, Australia, using the native Murray rainbowfish ( Melanotaenia fluviatilis )

The contamination of major continental river systems by endocrine-active chemicals (EACs) derived from the discharge of wastewater treatment plant (WWTP) effluents can affect human and ecosystem health. As part of a long-term effort to develop a native fish model organism for assessment of endocrine disruption in Australia's largest watershed, the Murray-Darling River Basin, the present study evaluated endocrine disruption in adult males of the native Australian Murray rainbowfish ( Melanotaenia fluviatilis ) exposed to effluent from an activated sludge WWTP and water from the Murray River during a 28-d, continuous-flow, on-site experiment. Analysis of the WWTP effluent and river water detected estrone and 17β-estradiol at concentrations up to approximately 25 ng L −1 . Anti-estrogenicity of effluent samples was detected in vitro using yeast-based bioassays (yeast estrogen screen) throughout the experiment, but estrogenicity was limited to the first week of the experiment. Histological evaluation of the testes indicated significant suppression of spermatogenesis by WWTP effluent after 28 d of exposure. Plasma vitellogenin concentrations and expression of vitellogenin messenger RNA in liver were not significantly affected by exposure to WWTP effluent. The combination of low contaminant concentrations in the WWTP effluent, limited endocrine disrupting effects in the Murray rainbowfish, and high in-stream dilution factors (>99%) suggest minimal endocrine disruption impacts on native Australian fish in the Murray River downstream from the WWTP outfall.

Upper Murray River↗

Observed and predicted reproduction of Ceriodaphnia dubia exposed to chloride, sulfate, and bicarbonate

Chronic toxicities of Cl - , SO 4 2- , and HCO 3 - to Ceriodaphnia dubia were evaluated in low- and moderate-hardness waters using a three-brood reproduction test method. Toxicity tests of anion mixtures were used to determine interaction effects and to produce models predicting C. dubia reproduction. Effluents diluted with low- and moderate-hardness waters were tested with animals acclimated to low- and moderate-hardness conditions to evaluate the models and to assess the effects of hardness and acclimation. Sulfate was significantly less toxic than Cl - and HCO 3 - in both types of water. Chloride and HCO 3 - toxicities were similar in low-hardness water, but HCO 3 - was the most toxic in moderate-hardness water. Low acute-to-chronic ratios indicate that toxicities of these anions will decrease quickly with dilution. Hardness significantly reduced Cl - and SO 4 2- toxicity but had little effect on HCO 3 - . Chloride toxicity decreased with an increase in Na + concentration, and CO 3 - toxicity may have been reduced by the dissolved organic carbon in effluent. Multivariate models using measured anion concentrations in effluents with low to moderate hardness levels provided fairly accurate predictions of reproduction. Determinations of toxicity for several effluents differed significantly depending on the hardness of the dilution water and the hardness of the water used to culture test animals. These results can be used to predict the contribution of elevated anion concentrations to the chronic toxicity of effluents; to identify effluents that are toxic due to contaminants other than Cl - , SO 4 2- , and HCO 3 - ; and to provide a basis for chemical substitutions in manufacturing processes.

Environmental Toxicology and Chemistry↗

Relative sensitivity of an amphipod Hyalella azteca , a midge Chironomus dilutus , and a unionid mussel Lampsilis siliquoidea to a toxic sediment

The objective of the present study was to evaluate the relative sensitivity of test organisms in exposures to dilutions of a highly toxic sediment contaminated with metals and organic compounds. One dilution series was prepared using control sand (low total organic carbon [TOC; <0.1%, low binding capacity for contaminants]) and a second dilution series was prepared using control sediment from West Bearskin Lake, Minnesota, USA (high TOC [∼10% TOC, higher binding capacity for contaminants]). Test organisms included an amphipod ( Hyalella azteca ; 10-d and 28-d exposures), a midge ( Chironomus dilutus ; 20-d and 48-d exposures started with <1-h-old larvae, and 13-d and 48-d exposures started with 7-d-old larvae), and a unionid mussel ( Lampsilis siliquoidea ; 28-d exposures). Relative species sensitivity depended on the toxicity endpoint and the diluent. All 3 species were more sensitive in sand dilutions than in West Bearskin Lake sediment dilutions. The <1-h-old C. dilutus were more sensitive than 7-d-old C. dilutus , but replicate variability was high in exposures started with the younger midge larvae. Larval biomass and adult emergence endpoints of C. dilutus exhibited a similar sensitivity. Survival, weight, and biomass of H. azteca were more sensitive endpoints in 28-d exposures than in 10-d exposures. Weight and biomass of L. siliquoidea were sensitive endpoints in both sand and West Bearskin Lake sediment dilutions. Metals, ammonia, oil, and other organic contaminants may have contributed to the observed toxicity.

Environmental Toxicology and Chemistry↗

Sensitivity of shovelnose sturgeon ( Scaphirhynchus platorynchus ) and pallid sturgeon ( S. albus ) early life stages to 3,30,4,40,5-pentachlorobiphenyl and 2,3,7,8-tetrachlorodibenzo-p-dioxin exposure

Concern exists that polychlorinated biphenyls (PCBs) may be contributing to the current decline of shovelnose sturgeon ( Scaphirhynchus platorynchus ) and the US federally endangered pallid sturgeon ( Scaphirhynchus albus ). Waterborne exposures with newly fertilized eggs were used to assess developmental and morphological effects of 2 of the most potent aryl hydrocarbon receptor (AhR) agonists, 3,3′,4,4′,5-pentachlorobiphenyl (PCB-126) and 2,3,7,8-tetrachlorodibenzo- p -dioxin (TCDD), on early life stage shovelnose and pallid sturgeon. No dose-related effects of PCB-126 were observed on percent development or hatch in either species at concentrations as high as 1711 ng/g egg. Effects of TCDD on percent development were not assessed in shovelnose sturgeon. However, percent development was not affected by TCDD in pallid sturgeon, and percent hatch was unaffected by TCDD doses as high as 60 ng/g egg to 81 ng/g egg in either species. Morphological pathologies such as yolk sac edema and craniofacial deformities were typical of AhR agonist exposure and were similar in both species. Calculated PCB-126 50% lethal dose (LD50, 95% fiducial limits) values were 196 ng/g egg (188–203 ng/g) for shovelnose and 159 ng/g egg (122–199 ng/g) for pallid sturgeon. Likewise, calculated TCDD LD50 values were 13 ng/g egg (11–15 ng/g) for shovelnose and 12 ng/g egg (10–14 ng/g) for pallid sturgeon. These LD50 values are among the highest recorded in early life stage fish, suggesting that early life stage Scaphirhynchus sturgeon may be comparatively insensitive to AhR agonists.

Environmental Toxicology and Chemistry↗

Evaluation of the short term 12 hour toxicity of 3-trifluoromethyl-4-nitrophenol (TFM) to multiple life stages of Venustaconcha ellipsiformis and Epioblasma triquetra and its host fish ( Percina caprodes )

The present study evaluated the risk of 12-h exposures of the lampricide 3-trifluoromethyl-4-nitrophenol (TFM) to multiple life stages of the federally endangered snuffbox ( Epioblasma triquetra ) and its primary host fish the common logperch ( Percina caprodes ) as well as a surrogate to the snuffbox, the ellipse ( Venustaconcha ellipsiformis ). Life stages examined included free glochidia, 1-wk juveniles, and adults of the ellipse; free glochidia, glochidia on host fish, and 1-wk juveniles of the snuffbox; and adult logperch. Larval sea lampreys were also tested alongside adult ellipse and logperch for direct comparison. Survival exceeded 82% among all life stages in both mussel species at levels up to 1.8 times what would be applied during treatments, suggesting that routine sea lamprey control operations would not adversely affect mussels. However, substantial mortality of adult logperch was observed at TFM concentrations typically applied to streams, and loss of host fish could adversely affect snuffbox reproduction. In addition, TFM had no significant effect on the number of glochidia that metamorphosed on adult logperch. Although the snuffbox is not likely to be acutely affected from sea lamprey control operations, mitigation efforts to minimize impacts to the host fish should be considered.

Environmental Toxicology and Chemistry↗

Influence of a chlor-alkali superfund site on mercury bioaccumulation in periphyton and low-trophic level fauna

In Berlin, New Hampshire, USA, the Androscoggin River flows adjacent to a former chlor-alkali facility that is a US Environmental Protection Agency Superfund site and source of mercury (Hg) to the river. The present study was conducted to determine the fate and bioaccumulation of methylmercury (MeHg) to lower trophic-level taxa in the river. Surface sediment directly adjacent to the source showed significantly elevated MeHg (10&ndash;40&times; increase, mean&thinsp;&plusmn;&thinsp;standard deviation [SD]: 20.1&thinsp;&plusmn;&thinsp;24.8 ng g &ndash;1 dry wt) and total mercury (THg; 10&ndash;30&times; increase, mean&thinsp;&plusmn;&thinsp;SD: 2045&thinsp;&plusmn;&thinsp;2669 ng g &ndash;1 dry wt) compared with all other reaches, with sediment THg and MeHg from downstream reaches elevated (3&ndash;7&times; on average) relative to the reference (THg mean&thinsp;&plusmn;&thinsp;SD: 33.5&thinsp;&plusmn;&thinsp;9.33 ng g &ndash;1 dry wt; MeHg mean&thinsp;&plusmn;&thinsp;SD: 0.52&thinsp;&plusmn;&thinsp;0.21&thinsp;ng g &ndash;1 dry wt). Water column THg concentrations adjacent to the point source for both particulate (0.23&thinsp;ng L &ndash;1 ) and dissolved (0.76&thinsp;ng L &ndash;1 ) fractions were 5-fold higher than at the reference sites, and 2-fold to 5-fold higher than downstream. Methylmercury production potential of periphyton material was highest (2&ndash;9&thinsp;ng g &ndash;1 d &ndash;1 dry wt) adjacent to the Superfund site; other reaches were close to or below reporting limits (0. 1&thinsp;ng g &ndash;1 d &ndash;1 dry wt). Total Hg and MeHg bioaccumulation in fauna was variable across sites and taxa, with no clear spatial patterns downstream of the contamination source. Crayfish, mayflies, and shiners showed a weak positive relationship with porewater MeHg concentration.

New Hampshire↗

Influence of in ovo mercury exposure, lake acidity, and other factors on common loon egg and chick quality in Wisconsin

A field study was conducted in Wisconsin (USA) to characterize in ovo mercury (Hg) exposure in common loons (Gavia immer). Total Hg mass fractions ranged from 0.17 mg/g to 1.23mg/g wet weight in eggs collected from nests on lakes representing a wide range of pH (5.0&ndash;8.1) and were modeled as a function of maternal loon Hg exposure and egg laying order. Blood total Hg mass fractions in a sample of loon chicks ranged from 0.84ug/g to 3.86 ug/g wet weight at hatch. Factors other than mercury exposure that may have persistent consequences on development of chicks from eggs collected on low-pH lakes (i.e., egg selenium, calcium, and fatty acid mass fractions) do not seem to be contributing to reported differences in loon chick quality as a function of lake pH. However, it was observed that adult male loons holding territories on neutral-pH lakes were larger on average than those occupying territories on low-pH lakes. Differences in adult body size of common loons holding territories on neutral-versus low-pH lakes may have genetic implications for differences in lake-source-related quality (i.e., size) in chicks. The tendency for high in ovo Hg exposure and smaller adult male size to co-occur in low-pH lakes complicates the interpretation of the relative contributions of each to resulting chick quality.

Wisconsin↗

Development of a new toxic-unit model for the bioassessment of metals in streams

Two toxic-unit models that estimate the toxicity of trace-metal mixtures to benthic communities were compared. The chronic criterion accumulation ratio (CCAR), a modification of biotic ligand model (BLM) outputs for use as a toxic-unit model, accounts for the modifying and competitive influences of major cations (Ca 2+ , Mg 2+ , Na + , K + , H + ), anions (HCO 3 − , CO 3 2− ,SO 4 2− , Cl − , S 2− ) and dissolved organic carbon (DOC) in determining the free metal ion available for accumulation on the biotic ligand. The cumulative criterion unit (CCU) model, an empirical statistical model of trace-metal toxicity, considers only the ameliorative properties of Ca 2+ and Mg 2+ (hardness) in determining the toxicity of total dissolved trace metals. Differences in the contribution of a metal (e.g., Cu, Cd, Zn) to toxic units as determined by CCAR or CCU were observed and attributed to how each model incorporates the influences of DOC, pH, and alkalinity. Akaike information criteria demonstrate that CCAR is an improved predictor of benthic macroinvertebrate community metrics as compared with CCU. Piecewise models depict great declines (thresholds) in benthic macroinvertebrate communities at CCAR of 1 or more, while negative changes in benthic communities were detected at a CCAR of less than 1. We observed a 7% reduction in total taxa richness and a 43% decrease in Heptageniid abundance between background (CCAR = 0.1) and the threshold of chronic toxicity on the basis of continuous chronic criteria (CCAR = 1). In this first application of the BLM as a toxic-unit model, we found it superior to CCU.

Environmental Toxicology and Chemistry↗

Mercury and selenium accumulation in the Colorado River food web, Grand Canyon, USA

Mercury (Hg) and selenium (Se) biomagnify in aquatic food webs and are toxic to fish and wildlife. The authors measured Hg and Se in organic matter, invertebrates, and fishes in the Colorado River food web at sites spanning 387 river&thinsp;km downstream of Glen Canyon Dam (AZ, USA). Concentrations were relatively high among sites compared with other large rivers (mean wet wt for 6 fishes was 0.17&ndash;1.59&thinsp;&mu;g&thinsp;g &ndash;1 Hg and 1.35&ndash;2.65&thinsp;&mu;g&thinsp;g &ndash;1 Se), but consistent longitudinal patterns in Hg or Se concentrations relative to the dam were lacking. Mercury increased (slope&thinsp;=&thinsp;0.147) with &delta; 15 N, a metric of trophic position, indicating biomagnification similar to that observed in other freshwater systems. Organisms regularly exceeded exposure risk thresholds for wildlife and humans (6&ndash;100% and 56&ndash;100% of samples for Hg and Se, respectfully, among risk thresholds). In the Colorado River, Grand Canyon, Hg and Se concentrations pose exposure risks for fish, wildlife, and humans, and the findings of the present study add to a growing body of evidence showing that remote ecosystems are vulnerable to long-range transport and subsequent bioaccumulation of contaminants. Management of exposure risks in Grand Canyon will remain a challenge, as sources and transport mechanisms of Hg and Se extend far beyond park boundaries. Environ Toxicol Chem 2015;9999:1&ndash;10

Arizona↗

In Response: Biological arguments for selecting effect sizes in ecotoxicological testing—A governmental perspective

Criticisms of the uses of the no-observed-effect concentration (NOEC) and the lowest-observed-effect concentration (LOEC) and more generally the entire null hypothesis statistical testing scheme are hardly new or unique to the field of ecotoxicology [ 1-4 ]. Among the criticisms of NOECs and LOECs is that statistically similar LOECs (in terms of p value) can represent drastically different levels of effect. For instance, my colleagues and I found that a battery of chronic toxicity tests with different species and endpoints yielded LOECs with minimum detectable differences ranging from 3% to 48% reductions from controls [ 5 ].

Environmental Toxicology and Chemistry↗

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

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

New York↗

Complex mixtures, complex responses: Assessing pharmaceutical mixtures using field and laboratory approaches

Pharmaceuticals are present in low concentrations (<100&thinsp;ng/L) in most municipal wastewater effluents but may be elevated locally because of factors such as input from pharmaceutical formulation facilities. Using existing concentration data, the authors assessed pharmaceuticals in laboratory exposures of fathead minnows ( Pimephales promelas ) and added environmental complexity through effluent exposures. In the laboratory, larval and mature minnows were exposed to a simple opioid mixture (hydrocodone, methadone, and oxycodone), an opioid agonist (tramadol), a muscle relaxant (methocarbamol), a simple antidepressant mixture (fluoxetine, paroxetine, venlafaxine), a sleep aid (temazepam), or a complex mixture of all compounds. Larval minnow response to effluent exposure was not consistent. The 2010 exposures resulted in shorter exposed minnow larvae, whereas the larvae exposed in 2012 exhibited altered escape behavior. Mature minnows exhibited altered hepatosomatic indices, with the strongest effects in females and in mixture exposures. In addition, laboratory-exposed, mature male minnows exposed to all pharmaceuticals (except the selective serotonin reuptake inhibitor mixture) defended nest sites less rigorously than fish in the control group. Tramadol or antidepressant mixture exposure resulted in increased splenic T lymphocytes. Only male minnows exposed to whole effluent responded with increased plasma vitellogenin concentrations. Female minnows exposed to pharmaceuticals (except the opioid mixture) had larger livers, likely as a compensatory result of greater prominence of vacuoles in liver hepatocytes. The observed alteration of apical endpoints central to sustaining fish populations confirms that effluents containing waste streams from pharmaceutical formulation facilities can adversely impact fish populations but that the effects may not be temporally consistent. The present study highlights the importance of including diverse biological endpoints spanning levels of biological organization and life stages when assessing contaminant interactions.

Environmental Toxicology and Chemistry↗

Acute and chronic toxicity of sodium sulfate to four freshwater organisms in water-only exposures

The acute and chronic toxicity of sulfate (tested as sodium sulfate) was determined in diluted well water (hardness of 100 mg/L and pH 8.2) with a cladoceran ( Ceriodaphnia dubia ; 2-d and 7-d exposures), a midge ( Chironomus dilutus ; 4-d and 41-d exposures), a unionid mussel (pink mucket, Lampsilis abrupta ; 4-d and 28-d exposures), and a fish (fathead minnow, Pimephales promelas ; 4-d and 34-d exposures). Among the 4 species, the cladoceran and mussel were acutely more sensitive to sulfate than the midge and fathead minnow, whereas the fathead minnow was chronically more sensitive than the other 3 species. Acute-to-chronic ratios ranged from 2.34 to 5.68 for the 3 invertebrates but were as high as 12.69 for the fish. The fathead minnow was highly sensitive to sulfate during the transitional period from embryo development to hatching in the diluted well water, and thus, additional short-term (7- to 14-d) sulfate toxicity tests were conducted starting with embryonic fathead minnow in test waters with different ionic compositions at a water hardness of 100 mg/L. Increasing chloride in test water from 10 mg Cl/L to 25 mg Cl/L did not influence sulfate toxicity to the fish, whereas increasing potassium in test water from 1mg K/L to 3mg K/L substantially reduced the toxicity of sulfate. The results indicate that both acute and chronic sulfate toxicity data, and the influence of potassium on sulfate toxicity to fish embryos, need to be considered when environmental guidance values for sulfate are developed or refined.

Environmental Toxicology and Chemistry↗

Accumulation of current-use and organochlorine pesticides in crab embyros from northern California, USA

Invertebrates have long been used as resident sentinels for assessing ecosystem health and productivity. The shore crabs, Hemigrapsus oregonensis and Pachygrapsus crassipes, are abundant in estuaries and beaches throughout northern California, USA and have been used as indicators of habitat conditions in several salt marshes. The overall objectives of the present study were to conduct a lab-based study to test the accumulation of current-use pesticides, validate the analytical method and to analyze field-collected crabs for a suite of 74 current-use and legacy pesticides. A simple laboratory uptake study was designed to determine if embryos could bioconcentrate the herbicide molinate over a 7-d period. At the end of the experiment, embryos were removed from the crabs and analyzed by gas chromatography/mass spectrometry. Although relatively hydrophilic (log K(OW) of 2.9), molinate did accumulate with an estimated bioconcentration factor (log BCF) of approximately 2.5. Following method validation, embryos were collected from two different Northern California salt marshes and analyzed. In field-collected embryos 18 current-use and eight organochlorine pesticides were detected including synthetic pyrethroids and organophosphate insecticides, as well as DDT and its degradates. Lipid-normalized concentrations of the pesticides detected in the field-collected crab embryos ranged from 0.1 to 4 ppm. Pesticide concentrations and profiles in crab embryos were site specific and could be correlated to differences in land-use practices. These preliminary results indicate that embryos are an effective sink for organic contaminants in the environment and have the potential to be good indicators of ecosystem health, especially when contaminant body burden analyses are paired with reproductive impairment assays.

California↗

Disparity between state fish consumption advisory systems for methylmercury and US Environmental Protection Agency recommendations: A case study of the South Central United States

Fish consumption advisories are used to inform citizens in the United States about noncommercial game fish with hazardous levels of methylmercury (MeHg). The US Environmental Protection Agency (USEPA) suggests issuing a fish consumption advisory when concentrations of MeHg in fish exceed a human health screening value of 300 ng/g. However, states have authority to develop their own systems for issuing fish consumption advisories for MeHg. Five states in the south central United States (Arkansas, Louisiana, Mississippi, Oklahoma, and Texas) issue advisories for the general human population when concentrations of MeHg exceed 700 ng/g to 1000 ng/g. The objective of the present study was to estimate the increase in fish consumption advisories that would occur if these states followed USEPA recommendations. The authors used the National Descriptive Model of Mercury in Fish to estimate the mercury concentrations in 5 size categories of largemouth bass–equivalent fish at 766 lentic and lotic sites within the 5 states. The authors found that states in this region have not issued site‐specific fish consumption advisories for most of the water bodies that would have such advisories if USEPA recommendations were followed. One outcome of the present study may be to stimulate discussion between scientists and policy makers at the federal and state levels about appropriate screening values to protect the public from the health hazards of consuming MeHg‐contaminated game fish.

Arkansas, Louisiana, Mississippi, Oklahoma, Texas↗

Landfill leachate as a mirror of today's disposable society: Pharmaceuticals and other contaminants of emerging concern in final leachate from landfills in the conterminous United States

Final leachates (leachate after storage or treatment processes) from 22 landfills in 12 states were analyzed for 190 pharmaceuticals and other contaminants of emerging concern (CECs), which were detected in every sample, with the number of CECs ranging from 1 to 58 (median&thinsp;=&thinsp;22). In total, 101 different CECs were detected in leachate samples, including 43 prescription pharmaceuticals, 22 industrial chemicals, 15 household chemicals, 12 nonprescription pharmaceuticals, 5 steroid hormones, and 4 animal/plant sterols. The most frequently detected CECs were lidocaine (91%, local anesthetic), cotinine (86%, nicotine degradate), carisoprodol (82%, muscle relaxant), bisphenol A (77%, component of plastics and thermal paper), carbamazepine (77%, anticonvulsant), and N,N-diethyltoluamide (68%, insect repellent). Concentrations of CECs spanned 7 orders of magnitude, ranging from 2.0&thinsp;ng/L (estrone) to 17&thinsp;200&thinsp;000&thinsp;ng/L (bisphenol A). Concentrations of household and industrial chemicals were the greatest (&sim;1000-1&thinsp;000&thinsp;000&thinsp;ng/L), followed by plant/animal sterols (&sim;1000-100&thinsp;000&thinsp;ng/L), nonprescription pharmaceuticals (&sim;100-10&thinsp;000&thinsp;ng/L), prescription pharmaceuticals (&sim;10-10&thinsp;000&thinsp;ng/L), and steroid hormones (&sim;10-100&thinsp;ng/L). The CEC concentrations in leachate from active landfills were significantly greater than those in leachate from closed, unlined landfills (p&thinsp;=&thinsp;0.05). The CEC concentrations were significantly greater (p&thinsp;<&thinsp;0.01) in untreated leachate compared with treated leachate. The CEC concentrations were significantly greater in leachate disposed to wastewater treatment plants from modern lined landfills than in leachate released to groundwater from closed, unlined landfills (p&thinsp;=&thinsp;0.04). The CEC concentrations were significantly greater (p&thinsp;=&thinsp;0.06) in the fresh leachate (leachate before storage or treatment) reported in a previous study compared with the final leachate sampled for the present study.

Arkansas, Florida, Iowa, Kentucky, Maine, Minnesot↗

Assessing variability in chemical acute toxicity of unionid mussels: Influence of intra- and inter-laboratory testing, life stage, and species

We developed a toxicity database for unionid mussels to examine the extent of intra- and inter-laboratory variability in acute toxicity tests with mussel larvae (glochidia) and juveniles; the extent of differential sensitivity of the two life stages; and the variation in sensitivity among commonly tested mussels ( Lampsilis siliquoidea , Utterbackia imbecillis , Villosa iris ), commonly tested cladocerans ( Daphnia magna , Ceriodaphnia dubia ) and fish ( Oncorhynchus mykiss , Pimephales promelas , Lepomis macrochirus ). The results of these analyses indicate intra-laboratory variability for median effect concentrations (EC50) averaged about 2 fold for both life stages, while inter-laboratory variability averaged 3.6 fold for juvenile mussels and 6.3 fold for glochidia. The EC50s for juveniles and glochidia were within a factor of 2 of each other for 50% of paired records across chemicals, with juveniles more sensitive than glochidia by more than 2 fold for 33% of the comparisons made between life stages. There was a high concurrence of the sensitivity of commonly tested L. siliquoidea , U. imbecillis , and V. iris to that of other mussels. However, this concurrence decreases as the taxonomic distance of the commonly tested cladocerans and fish to mussels increases. The compiled mussel database and determination of data variability will advance risk assessments by including more robust species sensitivity distributions, interspecies correlation estimates, and availability of taxon-specific empirically derived application factors for risk assessment.

Environmental Toxicology and Chemistry↗

Egg laying sequence influences egg mercury concentrations and egg size in three bird species: Implications for contaminant monitoring programs

Bird eggs are commonly used in contaminant monitoring programs and toxicological risk assessments, but intra-clutch variation and sampling methodology could influence interpretability. We examined the influence of egg laying sequence on egg mercury concentrations and burdens in American avocets, black-necked stilts, and Forster's terns. The average decline in mercury concentrations between the first and last egg laid was 33% for stilts, 22% for terns, and 11% for avocets, and most of this decline occurred between the first and second eggs laid (24% for stilts, 18% for terns, and 9% for avocets). Trends in egg size with egg laying order were inconsistent among species and overall differences in egg volume, mass, length, and width were <3%. We summarized the literature and, among 17 species studied, mercury concentrations generally declined by 16% between the first and second eggs laid. Despite the strong effect of egg laying sequence, most of the variance in egg mercury concentrations still occurred among clutches (75%-91%) rather than within clutches (9%-25%). Using simulations, we determined that to accurately estimate a population's mean egg mercury concentration using only a single random egg from a subset of nests, it would require sampling >60 nests to represent a large population (10% accuracy) or &ge;14 nests to represent a small colony that contained <100 nests (20% accuracy).

Environmental Toxicology and Chemistry↗