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Alternative tissue analysis method developed for organochlorine contaminants in aquatic organisms

The exposure of aquatic life to organochlorine contaminants has been investigated during the past two decades because of human and ecosystem health concerns related to the bioaccumulation of hazardous, lipophilic substances. The toxic effects of polychlorodibenzo-12-dioxins and polychlorinated biphenyls (PCBs) are well known, and recent evidence also suggests that low level exposure to lipophilic organochlorines may interfere with normal development during sensitive early life history stages (Hileman 1993). As the use of lipophilic organochlorines, such as DDT, in third world countries continues and with the purported global cycling (Bidleman and Olney 1974; Tanabe et al. 1983) and food chain accumulation (Thomann and Connoly 1984, Thomann 1989) of persistent organochlodnes, the occurrence of these compounds in aquatic organisms is a critical global environmental issue. An understanding of the fate of organochlorines in the environment clearly remains an extremely important subject related to water quality. The U.S. Geological Survey (USGS) has recently gained congressional approval in the United States to track nation wide trends in water quality through the establishment of the National Water Quality Assessment Program (NAWQA). Among the goals defined by NAWQA, aquatic organisms, including fish, shellfish, and plants, collected from major drainage basins will be analyzed for, along with other contaminants, polychlorinated biphenyls (PCBs), organochlorine insecticides, and chlorobenzenes (Crawford and Luoma 1992). The purpose of this report is to present quality assurance data obtained from the development of a PCB, chlorobenzene, and organochlorine insecticide tissue analysis method in support of NAWQA and other large-scale water quality programs conducted through our laboratory.

Bulletin of Environmental Contamination and Toxico↗

Interactive effects of selenium, methionine, and dietary protein on survival, growth, and physiology in mallard ducklings

Concentrations of over 100 ppm (100 mg/kg) selenium (Se) have been found in aquatic food chains associated with irrigation drainwater. Both quantity and composition of dietary protein for wild ducklings may vary in selenium-contaminated environments. Day-old mallard (Anas platyrhynchos) ducklings received one of the following diets containing 22% protein: unsupplemented (controls), 15 ppm Se (as selenomethionine), 60 ppm Se, methionine supplemented, 15 ppm Se with methionine supplement, or 60 ppm Se with methionine supplement. In a second concurrent experiment the above sequence was repeated with a protein-restricted (11%) but isocaloric diet. In a third concurrent experiment all ducklings received 44% protein with 0, 15, or 60 ppm Se added. After 4 weeks, blood and tissue samples were collected for biochemical and histological examination. With 22% protein and 60 ppm Se in the diet, duckling survival and growth was reduced and histopathological lesions of the liver occurred. Antagonistic interactive effects occurred between supplementary methionine and Se, including complete to partial alleviation of the following Se effects by methionine: mortality, hepatic lesions, and altered glutathione and thiol status. With 11% protein, growth of controls was less than that with 22% protein, Se (60 ppm) caused 100% mortality, and methionine supplementation, although protective afforded less protection than it did with 22% protein. With 44% protein, ducklings experienced physiological stress, and Se was more toxic than with methionine-supplemented 22% protein. These findings suggest the potential for antagonistic effects of Se, methionine, and protein on duckling survival and physiology.

Archives of Environmental Contamination and Toxico↗

Effects of 3,3′,4,4′,5-pentachlorobiphenyl (PCB 126) and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) injected into the yolks of chicken (Gallus domesticus) eggs prior to incubation

The yolks of White Leghorn chicken ( Gallus domesticus ) eggs were injected prior to incubation with either 3,3′,4,4′,5- pentachlorobiphenyl (PCB 126) at doses ranging from 0.1 to 12.8 μg/kg egg or 2,3,7,8-tetrachlorodibenzo- p -dioxin (TCDD) at doses ranging from 0.04 to 0.64 μg/kg egg. Chicks were subjected to necropsy within 24 h of hatching. The brain, bursa, heart, liver, and spleen were removed and weighed. Assessment of the rate of hatching indicated an LD 50 ±S.E. of 2.3±0.19 μg/kg egg (7.1±0.58 nmol/kg egg) for PCB 126 and 0.15±0.012 μg/kg egg (0.47±0.037 nmol/kg egg) for TCDD. No significant differences in the incidence of developmental abnormalities (structural defects and edema) were observed in TCDD-exposed embryos, while PCB 126 caused significantly more developmental abnormalities at 3.2, 6.4, and 12.8 μg/kg egg than the vehicle control. PCB 126 caused lower hatchling weights and greater relative brain, heart, and liver weights when compared to the vehicle control group at a dose of 3.2 μg/kg egg which is greater than the LD 50 . TCDD at 0.08 μg/kg egg caused relative bursa weights to be less than those of the vehicle control. A toxic equivalency factor (TEF) of 0.07 was determined for PCB 126 in relation to TCDD based on overt lethality.

Archives of Environmental Contamination and Toxico↗

Dietary exposure of mink to carp from Saginaw Bay, Michigan. 1. Effects on reproduction and survival, and the potential risks to wild mink populations

Carp ( Cyprinus carpio ) collected from Saginaw Bay, Michigan, containing 8.4 mg total polychlorinated biphenyls (PCBs)/kg and 194 ng of 2,3,7,8-tetrachloro-dibenzo- p -dioxin equivalents (TEQs)/kg, were substituted for marine fish at levels of 0, 10, 20, or 40% in the diets of adult ranch mink ( Mustela vison ). The diets, containing 0.015, 0.72, 1.53, and 2.56 mg PCBs/kg diet, or 1.03, 19.41, 40.02, and 80.76 ng TEQs/kg diet, respectively, were fed to mink prior to and throughout the reproductive period to evaluate the effects of a naturally-contaminated prey species on their survival and reproductive performance. The total quantities of PCBs ingested by the mink fed 0, 10, 20, or 40% carp over the 85-day treatment period were 0.34, 13.2, 25.3, and 32.3 mg PCBs/mink, respectively. The corresponding quantities of TEQs ingested by the mink over the same treatment period were 23, 356, 661, and 1,019 ng TEQs/mink, respectively. Consumption of feed by mink was inversely proportional to the PCB and TEQ content of the diet. The diets containing Saginaw Bay carp caused impaired reproduction and/or reduced survival of the kits. Compared to controls, body weights of kits at birth were significantly reduced in the 20 and 40% carp groups, and kit body weights and survival in the 10 and 20% carp groups were significantly reduced at three and six weeks of age. The females fed 40% carp whelped the fewest number of kits, all of which were stillborn or died within 24 hours. Lowest observable adverse effect levels (LOAEL) of 0.134 mg PCBs/kg body weight/day or 3.6 ng TEQs/kg body weight/day for adult female mink were determined. The potential effects of exposure of wild mink to contaminated Great Lakes fish were assessed by calculating “maximum allowable daily intakes” and “hazard indices” based on total concentrations of PCB residues in several species of Great Lakes fish and mink toxicity data derived from the study.

Archives of Environmental Contamination and Toxico↗

Environmental contaminants in redheads wintering in coastal Louisiana and Texas

Whole body and liver analyses indicated that wintering redheads (Aythya americana; n = 70) in coastal Louisiana (one site) and Texas (two sites) were relatively free of contamination with common trace elements, organochlorines, and hydrocarbons. Most trace elements, including As, Cr, Hg, Mg, Mn, Ni, Pb, Se, Sr, and Zn, were within background concentrations in livers; levels of B, Cd, Cu, and Fe were elevated in some specimens. Only one organochlorine, DDE, was detected in redhead carcasses, but its concentration was below reported toxic levels in waterfowl. Body burdens of aliphatic and aromatic hydrocarbons were generally low, but levels of pristane, total hydrocarbons, and the ratios of phytane:n-octadecane and pristane:n-heptadecane were indicative of possible chronic exposure to petroleum. Based on brain cholinesterase assays, redheads were not recently exposed to organophosphorous or carbamate pesticides. Of 30 elements or compounds tested for seasonal differences, only Se increased from early to late winter at one of the three sites. Eight of 57 contaminants differed among the three sites; no sex or age differences were found.

Louisiana, Texas↗

Main and interactive effects of arsenic and selenium on mallard reproduction and duckling growth and survival

Arsenic (As) and selenium (Se) occur together in high concentrations in the environment and can accumulate in aquatic plants and invertebrates consumed by waterfowl. Ninety-nine pairs of breeding mallards ( Anas platyrhynchos ) were fed diets supplemented with As (sodium arsenate) at 0, 25, 100, or 400 μg/g, in combination with Se (seleno-DL-methionine) at 0 or 10 μg/g, in a replicated factorial experiment. Ducklings produced were placed on the same treatment combination as their parents. Arsenic accumulated in adult liver and egg, reduced adult weight gain and liver weight, delayed the onset of egg laying, decreased whole egg weight, and caused eggshell thinning. Arsenic did not affect hatching success and was not teratogenic. In ducklings, As accumulated in the liver and reduced body weight, growth, and liver weight. Arsenic did not increase duckling mortality, but it did decrease overall duckling production. Selenium accumulated in adult liver and egg, was teratogenic, and decreased hatching success. Selenium did not affect adult weight, liver weight, survival, onset of egg laying, egg fertility, egg weight, or eggshell thickness. In ducklings, Se accumulated in the liver and reduced body weight and growth, and increased liver weight. Selenium increased duckling mortality and decreased overall duckling production. Antagonistic interactions between As and Se occurred whereby As reduced Se accumulation in liver and egg, and alleviated the effects of Se on hatching success and embryo deformities. It was demonstrated that As and Se, in the chemical forms and at the dietary levels administered in this study, can adversely affect mallard reproduction and duckling growth and survival, and that As can alleviate toxic effects of Se.

Archives of Environmental Contamination and Toxico↗

Biomonitoring of lead-contaminated Missouri streams with an assay for erythrocyte δ-aminolevulinic acid dehydratase activity in fish blood

The activity of the enzyme δ-aminolevulinic acid dehydratase (ALA-D) in erythrocytes has long been used as a biomarker of lead exposure in humans and waterfowl and, more recently, in fishes. The assay was tested for ALA-D activity in fishes from streams affected by lead in combination with other metals from lead-zinc mining and related activities. Fishes (mostly catostomids) were collected from sites affected by historic and current mining activities, and from sites considered to be unaffected by mining (reference sites). A group of potentially toxic elements was measured in blood and carcass samples of individual fish, as were ALA-D activity, total protein (TP), and hemoglobin (Hb) in blood. Concentrations of mining-related metals (lead, zinc, and cadmium) were significantly greater (P<0.05) in fish blood and carcass at sites affected by historic mining activities than at reference and active mining sites. When analyzed by multiple regression, ALA-D activity, Hb, and TP accounted for 66% of blood-lead and 69% of carcass-lead variability. Differences among species were small. ALA-D activity as a biomarker adequately distinguished sites affected by bioavailable environmental lead. Zinc was the only other metal that affected ALA-D activity; it appeared to ameliorate the inactivation of ALA-D by lead.

Archives of Environmental Contamination and Toxico↗

Organochlorine accumulation by sentinel mallards at the Winston-Thomas sewage treatment plant, Bloomington, Indiana

Farm-raised 12-month-old female mallards ( Anas platyrhynchos ) were released at the Winston-Thomas sewage treatment plant, Bloomington, Indiana. Five mallards were sacrificed at the start of the study and at approximately 10-day intervals through day 100. Concentrations of polychlorinated biphenyls (PCBs) in carcasses increased linearly with time of exposure and exceeded 16 μg/g wet weight by day 100; PCBs in breast muscle exceeded 3.9 μg/g by day 100. These PCB values are among the highest recorded for wild or sentinel waterfowl. PCB concentrations in breast muscle (26–523 μg/g lipid weight) were 50–1,000 times greater than human consumption guidelines for edible poultry in Canada (0.5 μg/g lipid weight) and 9–176 times greater than consumption guidelines for edible poultry in the United States (3.0 μg/g lipid weight). Additionally, PCB concentrations in carcass and breast muscle exceeded the threshold of the Great Lakes Sport Fish Consumption Advisory ‘do not eat’ category (1.9 μg/g wet weight) by day 20 and day 50, respectively. Hepatic cytochrome P450-associated monooxygenases including BROD (benzyloxyresorufin- O -dealkylase), EROD (ethoxyresorufin- O -dealkylase), and PROD (pentoxyresorufin- O -dealkylase) were induced over 5-fold compared to reference mallards. BROD, EROD, and PROD were each significantly correlated to total PCBs and to the toxicity of selected PCB congeners, relative to 2,3,7,8-tetrachlorodibenzo- p -dioxin.

Indiana↗

A survey of Pb, Cu, Zn, Cd, Cr, As, and Se in earthworms and soil from diverse sites

Earthworms and soils were collected from 20 diverse sites in Maryland, Pennsylvania, and Virginia, and were analyzed for Pb, Cu, Zn, Cd, Cr, As, and Se. Correlation coefficients relating concentrations of the elements in earthworms to concentrations in soil were low (−0.20< r <+0.57). Species differences and ecological variables contributed to the variability in concentrations of these elements in earthworms. The maximum concentrations of Pb (2100 ppm), Zn (1600 ppm), Cd (23 ppm) and Se (7.6 ppm) detected in earthworms were in the range reported to be toxic to animals fed diets containing these elements; however, even in the absence of any environmental contamination, some species of earthworms may contain high concentrations of Pb, Zn, and Se. Earthworms of the genus Eisenoides , for example, were exceptional in their ability to concentrate Pb. When earthworms are used as indicators of environmental contamination, it is important to identify the species, to report the soil characteristics, and to collect similar earthworms from very similar but uncontaminated soil.

Environmental Monitoring and Assessment↗

Contaminant exposure of willets feeding in agricultural drainages of the lower Rio Grande valley of south Texas

Willets ( Catoptrophorus semipalmatus ) were collected in June and August 1986 at the outlets of two agricultural drainages into the Lower Laguna Madre of South Texas and at two other Texas coastal sites. Mean liver concentration of arsenic was higher in August than June. Over 20% of the livers had arsenic concentrations elevated above a suggested background level of 5.0 ppm dry weight (DW), but concentrations (maximum 15 ppm) were below those associated with acute toxicity. Selenium concentration in livers varied from 2.3 to 8.3 ppm DW for all locations and represented background levels. Mercury concentrations in livers for all locations (means = 2.0 to 3.4, maximum 17 ppm DW) were below those associated with avian mortality and similar to levels found in other estuarine/marine birds. DDE in carcasses was higher in adults (mean = 1.0 ppm wet weight) than juveniles (0.2 ppm), and higher in August (1.0 ppm) than June (0.5 ppm); however, DDE concentrations were generally at background levels. Based on brain cholinesterase activity, willets were not recently exposed to organophosphate pesticides.

Environmental Monitoring and Assessment↗

Ambient air concentrations of PCDDs, PCDFs, coplanar PCBs, and PAHs at the Mississippi Sandhill Crane National Wildlife Refuge, Jackson County, Mississippi

Our objective was to determine the levels of selected airborne contaminants in ambient air at the Mississippi Sandhill Crane National Wildlife Refuge, Mississippi, that might be affecting the health of endangered cranes living there. Two high-volume air samplers were operated at separate locations on the Refuge during May–September 1991. The sampling media were micro-quartz filters in combination with polyurethane foam plugs. Composite bimonthly samples from each station were analyzed for polychlorinated dibenzo- p -dioxins (PCDDs), dibenzofurans (PCDFs), coplanar polychlorinated biphenyls (PCBs), and polycyclic aromatic hydrocarbons (PAHs). Overall, residue concentrations were low. The toxic PCDD isomer 2,3,7,8-tetra-CDD was not detected, nor was penta-CDD. There was no difference ( P >0.05) in residue concentrations between stations or over time and meteorological parameters were not correlated with residue concentrations. Because contaminant levels and patterns may differ seasonally, we recommend that air samples collected during winter months also be analyzed for these same chemical groups.

Environmental Monitoring and Assessment↗

Survey and evaluation of contaminants in earthworms and in soils derived from dredged material at confined disposal facilities in the Great Lakes region

Soils derived from dredged material were collected, together with earthworms from nine confined disposal facilities located in the Great Lakes Region. These samples were analyzed for 18 elements, 11 organochlorine pesticides, PCBs, and 24 polycyclic aromatic hydrocarbons. The concentrations detected in earthworms were evaluated in terms of their potential hazard to wildlife, which for the sake of the evaluation were assumed to prey entirely either on earthworms or on other soil invertebrates having similar concentrations. The soil concentrations (dry wt.) of the contaminants of greatest concern were <1.9 to 32 ppm Cd, <0.053 to 0.94 ppm Hg, 4.6 to 550 ppm Pb, and <0.1 to 1.0 ppm PCBs. The concentrations in earthworms (dry wt., ingested soil included) were as high as 91 ppm Cd, 1.6 ppm Hg, 200 ppm Pb, and 1.8 ppm PCBs. Based on laboratory toxicity studies of relatively sensitive species, and on concentration factors calculated from the earthworm and soil data, we estimated that lethal or serious sublethal effects on wildlife might be expected at concentrations of 10 ppm Cd, 3 ppm Hg, 670 ppm Pb, and 1.7 ppm PCBs in alkaline surface soils derived from dredged material. Concentrations of polycyclic aromatic hydrocarbons in earthworms were well below those in soil.

Environmental Monitoring and Assessment↗

Environmental contaminants in nonviable eggs of the endangered Mississippi sandhill crane (Grus canadensis pulla)

Our objectives were to determine if concentrations of environmental pollutants and microbial contamination in nonviable eggs of the endangered Mississippi sandhill crane ( Grus canadensis pulla ) contributed to egg failure. Six eggs collected in 1990 and four in 1991 contained only background levels of organochlorine pesticides and polychlorinated biphenyls (PCBs), and tests for microbial contamination were all negative. Two eggs contained late dead embryos, but neither revealed obvious abnormalities. Three eggs contained potentially harmful concentrations (23, 39, 146 pg/g, wet mass) of polychlorinated dibenzo- p -dioxins (PCDDs) and dibenzofurans (PCDFs), based on 2,3,7,8-tetrachlorodibenzo- p -dioxin equivalents (TCDD-EQ) for combined compounds. Because of the scarcity of material suitable for laboratory examination and the endangered status of the crane, we recommend that nonviable eggs continue to be monitored for toxic pollutants.

Mississippi↗

Reactive solute transport in acidic streams

Spatial and temporal profiles of Ph and concentrations of toxic metals in streams affected by acid mine drainage are the result of the interplay of physical and biogeochemical processes. This paper describes a reactive solute transport model that provides a physically and thermodynamically quantitative interpretation of these profiles. The model combines a transport module that includes advection-dispersion and transient storage with a geochemical speciation module based on MINTEQA2. Input to the model includes stream hydrologic properties derived from tracer-dilution experiments, headwater and lateral inflow concentrations analyzed in field samples, and a thermodynamic database. Simulations reproduced the general features of steady-state patterns of observed pH and concentrations of aluminum and sulfate in St. Kevin Gulch, an acid mine drainage stream near Leadville, Colorado. These patterns were altered temporarily by injection of sodium carbonate into the stream. A transient simulation reproduced the observed effects of the base injection.

Water, Air, & Soil Pollution↗

Accumulation of 14C-naphthalene in the tissues of redhead ducks fed oil-contaminated crayfish

Crayfish, artificially contaminated with 14 C-naphthalene-5% water-soluble fraction of No. 2 fuel oil, were force-fed to one-year-old redhead ducks to determine the accumulation of petroleum hydrocarbons. The relative distribution of carbon-14 activity in the gall bladder containing bile, and fat were similar, and significantly greater (P < 0.05) than the activity in the blood, brain, liver, and kidney. There was a significant increase (P < 0.05) in the disintegrations per minute per gram (dpm/g) in the blood, brain, kidney, and liver between days 1 and 3 of feeding, indicating a progressive accumulation of carbon-14 activity (naphthalene and presumably its metabolites). There was no significant effect of sex or the interaction of the duration of feeding and sex on carbon-14 activity in any of the tissues. The low daily dose of petroleum hydrocarbons (a total of approximately 1.25 mg/day) received by the ducks from the crayfish and the relatively short feeding regimen did not cause any overt signs of toxicity in the ducks.

Archives of Environmental Contamination and Toxico↗

Effect of pH on the accumulation kinetics of pentachlorophenol in goldfish

The kinetics of accumulation of pentachlorophenol (PCP) at various pH values were investigated to explore how pH-dependent accumulation might influence PCP toxicity. Goldfish (Carassius auratus) were exposed to 5 μg PCP/L in a static system buffered with 7.5 mM bicine or N,N -bis(2-hydroxyethyl)-2-aminoethane sulfonic acid (BES) at pH 7.0, 8.0, or 9.0. The amount of PCP in the fish, concentration of PCP in water, and the total amount of metabolites in the system were measured after exposure of fish from 1 to 96 hr. Equations for these variables based on a two compartment pharmacokinetic model were fitted simultaneously to the data using NONLIN, which uses an iterative nonlinear least squares technique. Uptake clearance, metabolic clearance, and apparent volume of distribution of PCP decreased as pH increased. The decrease in PCP accumulation with increased pH was not due solely to a pH-induced decrease in uptake. In addition, the distribution of PCP within the fish was altered by changes in the external pH. The pH-associated changes in distribution may have altered access of PCP to sites of metabolism, thereby altering the metabolic clearance. The pH-related changes in the pharmacokinetics of PCP resulted in a decrease in its bioconcentration factor with an increase in pH and account both for the decreased capacity of the fish to accumulate PCP and for its reduced LC50.

Archives of Environmental Contamination and Toxico↗

Survival of striped bass larvae and yearlings in relation to contaminants and water quality in the upper Chesapeake Bay

This study was designed to evaluate survival of striped bass yolk-sac larvae and yearlings at three locations in their natural spawning habitat in the upper Chesapeake Bay (Chesapeake and Delaware Canal) using “in-situ” chambers; correlate larval and yearling survival with the presence of 11 water quality parameters, 10 inorganic contaminants and 21 organic contaminants and assess histological effects on yearlings after exposure to habitat water. The cumulative percent survival ranged from 42–59.5% for striped bass larvae after 96 hr of exposure to habitat water during two experiments. Survival in control conditions during these two experiments was 77.5 and 80.5%. Data from previous studies have shown that mortality rates for wild yolk-sac larvae ranged from 7–19% per day. Daily mortality of larvae in the present study was 13–16%; therefore, suspected acutely harmful water quality or contaminant conditions affecting survival were not substantiated. All yearling striped bass survived 10 days of exposure to habitat water. Although habitat water was not acutely toxic, histological examination of surviving yearling striped bass indicated sublethal effects. Telangiectases (lamellar dilations) of the gills was reported for yearlings exposed to habitat water. This pathological change was not found in the controls.

Delaware, Maryland↗

Effects of oral doses of fluoride on nestling European starlings

Nestling European starlings ( Sturnus vulgaris ), raised and fed by free-living adults, were given daily oral doses of either distilled water, 193 mg sodium as Na 2 CO 3 per kg of body weight (sodium control group), or 6, 10, 13, 17, 23, 30, 40, 80, 160 mg of the fluoride ion as NaF in distilled water per kg of body weight (mg/kg). Dosing began when nestlings were 24–48 hr old and continued for 16 days. The 24-hr LD 50 of fluoride for day-old starlings was 50 mg/kg. The 16-day LD 50 was 17 mg/kg. The sodium control group did not differ from the water control group with respect to any of the measured variables. Growth rates were significantly reduced in the 13 and 17 mg of fluoride/kg groups; weights of birds given higher dose levels were omitted from growth comparisons because of high, fluoride-induced mortality. Although prefledging weights for the 10, 13, and 17 mg of fluoride/kg groups averaged 3.6 to 8.6% less than controls at 17 days, this difference was not significant. Feather and bone growth of the fluoride and control groups were not different, except for keel length measured at 17 days of age which averaged less in the fluoride groups. Liver and spleen weights were not affected by fluoride treatments. No histological damage related to fluoride treatments was found in liver, spleen, or kidney. The logarithm of bone fluoride and magnesium concentration increased with the logarithm of increasing fluoride treatment levels and were significantly correlated with each other. Fluoride treatments had no effect on percent calcium or phosphorus in bone or plasma alkaline phosphatase activity. Oral doses of fluoride appear to be more toxic than equivalent dietary levels. Most birds probably acquire fluoride through their diet. Therefore, the results of the study may overstimate the potential effects of fluorides on songbirds living in fluoride-contaminated environments.

Archives of Environmental Contamination and Toxico↗