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Effects of boron and selenium on mallard reproduction and duckling growth and survival

Boron (B) and selenium (Se) sometimes occur together in high concentrations in the environment and can accumulate in plants and invertebrates consumed by waterfowl. One hundred twenty-six pairs of breeding mallards (Anas platyrhynchos) were fed diets supplemented with B (as boric acid) at 0, 450, or 900 ppm, in combination with Se (as seleno-DL-methionine) at 0, 3.5, or 7 ppm, in a replicated factorial experiment. Ducklings produced received the same treatment combination as their parents. Boron and Se accumulated in adult liver, egg, and duckling liver. In adults, B and Se caused weight loss, and B decreased hemoglobin concentration, egg weight, and egg fertility. Both B and Se reduced hatching success and duckling weight, and B reduced duckling growth and duckling production, and caused several alterations in duckling liver biochemistry. Duckling survival was not reduced by B or Se, and neither B nor Se had histopathologic effects on adult or duckling liver, kidney, or spleen. There was little evidence of interaction between B and Se. This study demonstrated that B and Se, in the chemical forms and at the dietary levels administered in this study, can adversely affect mallard reproduction and duckling growth.

Environmental Toxicology and Chemistry↗

Sediment quality assessment studies of Tampa bay, Florida

A survey of the toxicity of sediments throughout the Tampa Bay estuary was performed as part of the National Oceanic and Atmospheric Administration's National Status and Trends Program. The objectives of the survey were to determine the spatial extent and severity of toxicity and to identify relationships between chemical contamination and toxicity. Three independent toxicity tests were performed: a 10-d amphipod survival test of the whole sediments with Ampelisca abdita , a sea urchin fertilization test of sediment pore water with Arbacia punctulata , and a 5-min Microtox® bioluminescence test with solvent extracts of the sediments. Seventy-three percent of the 165 undiluted sediment pore-water samples were significantly toxic relative to reference samples with the sea urchin fertilization test. In contrast, only 2% of the 165 samples were significantly toxic in the amphipod tests. The causes of toxicity were not determined. However, concentrations of numerous trace metals, pesticides, polychlorinated biphenyl (PCB) congeners, polycyclic aromatic hydrocarbons (PAHs), and ammonia were highly correlated with pore-water toxicity. Concentrations of many substances, especially total dichlorodiphenyltrichloroethanes (DDTs), endrin, total PCBs, certain PAHs, lead, and zinc, occurred at concentrations in the toxic samples that equaled or exceeded concentrations that have been previously associated with sediment toxicity.

Florida↗

Interlaboratory study of precision: Hyalella azteca and Chironomus tentans freshwater sediment toxicity assays

Standard 10-d whole-sediment toxicity test methods have recently been developed by the U.S. Environmental Protection Agency (EPA) for the amphipod Hyalella azteca and the midge Chironomus tentans . An interlaboratory evaluation of method precision was performed using a group of seven to 10 laboratories, representing government, academia, and environmental consulting firms. The test methods followed the EPA protocols for 4-d water-only reference toxicant (KCl) testing (static exposure) and for 10-d whole-sediment testing. Test sediments included control sediment, two copper-containing sediments, and a sediment contaminated primarily with polycyclic aromatic hydrocarbons. Reference toxicant tests resulted in H. azteca and C. tentans median lethal concentration (LC50) values with coefficents of variation (CVs) of 15.8 and 19.6%, respectively. Whole sediments which were moderately contaminated provided the best estimates of precision using CVs. Hyalella azteca and C. tentans tests in moderately contaminated sediments exhibited LC50 CVs of 38.9 and 13.5%, respectively. The CV for C. tentans growth was 31.9%. Only 3% (1 of 28) of samples exceeded acceptable interlaboratory precision limits for the H. azteca survival tests. No samples exceeded the intralaboratory precision limit for H. azteca or C. tentans survival tests. However, intralaboratory variability limits for C. tentans growth were exceeded by 80 and 100% of the laboratories for a moderately toxic and control sample, respectively. Interlaboratory variability limits for C. tentans survival were not exceeded by any laboratory. The results showed these test methods to have relatively low variance and acceptable levels of precision in interlaboratory comparisons.

Environmental Toxicology and Chemistry↗

Acute toxicity of three fire-retardant and two fire-suppressant foam formulations to the early life stages of rainbow trout ( Oncorhynchus mykiss )

Laboratory studies were conducted with five early life stages of rainbow trout, Oncorhynchus mykiss , to determine the acute toxicities of five fire-fighting chemical formulations in standardized soft and hard water. Eyed egg, embryo–larvae, swim-up fry, and 60- and 90-d posthatch juveniles were exposed to three fire retardants (Fire-Trol LCG-R, Fire-Trol GTS-R, and Phos-Chek D75-F) and two fire-suppressant foams (Phos-Chek WD-881 and Silv-Ex). Swim-up fry were generally the most sensitive life stage, whereas the eyed-egg was the least sensitive. Toxicity of fire-fighting formulations was greater in hard water than in soft water for all life stages tested with Fire-Trol GTS-R and Silv-Ex and for 90-d-old juveniles tested with Fire-Trol LCG-R. The fire-suppressant foams were more toxic than the fire retardants. The 96-h median lethal concentrations (LC50s) were ranked from the most toxic to the least toxic formulation as follows (ranges are the lowest and highest 96-h LC50 calculated for each formulation): Phos-Chek WD-881 (11–44 mg/L), Silv-Ex (11–78 mg/L), Phos-Chek D75-F (218–>3,600 mg/L), Fire-Trol GTS-R (207–>6,000 mg/L), and Fire-Trol LCG-R (872–>10,000 mg/L). Toxicity values suggest that accidental entry of fire-fighting chemicals into aquatic environments could adversely affect fish populations.

Environmental Toxicology and Chemistry↗

Small mammal populations in Maryland meadows during four years of herbicide (brominal®) applications

The herbicide Brominal® was applied at the recommended rate to one plot in each of three paired 0.6-ha plots; the other three plots were used as controls. Plots were sprayed once in the fall of 1988 and 1989 and twice in the spring of 1990 and 1991. Small mammals were trapped three times during each activity season (April-October) to obtain population estimates before and after spraying and in the spring preceding fall spraying or the fall following spring spraying. Population estimates immediately after spraying gave no evidence of direct mortality. By 1991, dicot vegetation on treated plots was suppressed and mean numbers of meadow voles (Microtus pennsylvanicus) were less than on control plots. Because meadow voles favor dicots over monocots in their diet, reduced availability of dicots may have been related to the smaller vole population estimates. Species diversity of small mammals was negatively correlated with size of vole populations, but was not different between treated and control plots. Brominal apparently induced opaque corneas in nine voles. The condition was found in two voles too small to have been conceived at the time of the last previous spray nearly 8 months earlier, suggesting exposure to residue alone.

Environmental Toxicology and Chemistry↗

A comparison of β-adrenoceptors and muscarinic cholinergic receptors in tissues of brown bullhead catfish ( Ameiurus nebulosus ) from the black river and old woman creek, Ohio

β-Adrenoceptors (βARs) and muscarinic cholinergic receptors were measured in brain, gill, and heart tissues of brown bullhead catfish exposed to polycyclic aromatic hydrocarbons in the Black River, Ohio, USA, and were compared to values from Old Woman Creek, Ohio, a reference site. A decreased number of βARs were found in the gill from Black River fish, possibly indicating a compensatory response subsequent to chemical stress.

Environmental Toxicology and Chemistry↗

A resource conservative procedure for comparison of dose-response relationships

The evaluation of effects of toxicants on a wildlife community can be complicated by varying responses among the community's constituent populations. Even within populations, considerable variability in dose-response relations may result from different avenues of exposure to the toxicant. Full-scale investigations of the dose-response relations among a variety of species and avenues of exposure can therefore be prohibitively expensive, whether this expense is measured by the number of experimental animals needed, by the human resources committed to the study, or by laboratory expenses. We propose an abbreviated protocol for investigations of multiple dose-response relations that is designed to limit these expenses. The protocol begins with the judicious choice of a baseline dose-response relation to be estimated by a full-scale study involving a minimum of five doses levels, with 10 subjects per dose level. This relation is then used as the basis for rapid screening of subsequent dose-response relations, which are compared to the baseline relation by testing for differences in the median effective dosages. These secondary studies can consist of as few as 14 animals exposed to the estimated median lethal concentration from the baseline study. We describe MS-DOS-compatible software available from the authors that can be used to analyze these data.

Environmental Toxicology and Chemistry↗

The role of water ventilation and sediment ingestion in the uptake of benzo[A]pyrene in gizzard shad ( Dorosoma cepedianum )

The objective of this study was to determine whether sediment ingestion or water ventilation was the primary route of uptake for benzo[ a ]pyrene (B a P) in the gizzard shad ( Dorosoma cepedianum ), a detritivorous fish. Two experiments were conducted in which fish were exposed to sediments spiked with 1 μg/g B a P. In the first experiment, fish were prevented from feeding by esophagus ligation. In the second experiment, 20 nonligated fish and 30 ligated fish were added to the aquarium. The nonligated fish roiled the water as they fed. Fish were collected 4, 8, 15, and 22 d after the experiments began. Gizzard shad metabolize B a P; therefore, the concentrations of B a P equivalents (parent B a P plus metabolite) were determined. Concentrations of B a P equivalents were significantly greater in the ligated fish in experiment 2 relative to those in experiment 1. In contrast, the concentration of B a P equivalents in the ligated fish in experiment 2 was not significantly different than that in the nonligated fish. Our results suggest that ventilation of turbid water may be a significant source of B a P for gizzard shad. Sediment ingestion, however, does not appear to significantly influence the total body concentration of B a P equivalents in gizzard shad.

Environmental Toxicology and Chemistry↗

A two-step experimental design for a sediment bioassay using growth of the amphipod Hyalella azteca for the test end point

We designed a sediment bioassay using 25% growth inhibition of Hyalella azteca as the end point. Hyalella azteca exhibits size-specific fecundity, so growth is a surrogate of reproductive production. We investigated density effects on growth to address whether crowding could affect test interpretation; amphipods in 14,000/m 2 exposures were 16 to 20% smaller than those at 7,000/m 2 . Using power analysis, we found that 20 to 25 samples are required to determine significance when α = 0.10 and 1 − β = 0.90. To minimize the need for laboratory resources, we designed a two-step (screening and confirmatory) bioassay, which we tested with field-collected sediments. The screening bioassay compared 11 sediments to a reference. Three sediments were “toxic” (significant growth inhibition when 1 − β = 0.66 and n = 5), five sediments were “nontoxic” (>90% of reference), and three sediments were “possibly toxic” (growth inhibition was insignificant). In the confirmatory bioassay, three possibly toxic and two nontoxic samples were reevaluated. Two were toxic (1 − β = 0.91 and n = 20), and the remaining four samples were nontoxic. In summary, five sediments were toxic and six sediments were nontoxic. The two-step analysis used minimal laboratory resources but maximized statistical power, where needed, to discriminate growth effects.

Environmental Toxicology and Chemistry↗

Ecological restructuring in experimental aquatic mesocosms due to the application of diflubenzuron

Diflubenzuron is a chitin-inhibiting insecticide that is registered for use in controlling a multitude of invertebrate pests. The selective mode of action of diflubenzuron provided us with a unique tool for examining the indirect responses of an experimental aquatic ecosystem following direct impacts at the primary consumer (invertebrate) trophic level. Twelve outdoor aquatic mesocosms (0.1 ha each) were randomly assigned to three treatments of diflubenzuron (four mesocosms/treatment): control, monthly (five total 10-μg/L applications), or biweekly (twice monthly; nine total 10-μg/L applications). Direct impacts on invertebrates (numbers and species composition of insects and zooplankton) and indirect impacts on fish (mortality, growth, and reproduction of bluegill sunfish and largemouth bass), phytoplankton (biomass and productivity), and water quality were measured over 16 weeks posttreatment. Both monthly and biweekly treatments of diflubenzuron decreased total numbers (fivefold reduction) and species richness (twofold reduction) and increased dominance (twofold increase) of zooplankton within 4 weeks posttreatment; Cladocerans, Copepods, and Rotifers were all sensitive. Diflubenzuron reduced species richness of emergent insects, which resulted in increased dominance by a few species; however, impacts on emergent insects were confounded by a treatment × time interaction. Direct reductions in invertebrate grazers caused indirect increases in algal biomass. Indirect effects on biomass (50% reduction) and individual weight (50% reduction) of juvenile bluegill occurred because of apparent decreases in invertebrate food resources. No statistically significant impacts were observed on adult bluegill or largemouth bass for the duration of the experiment. Results indicated that diflubenzuron had both direct and indirect impacts on the experimental aquatic ecosystems under the conditions tested.

Environmental Toxicology and Chemistry↗

Purification and immunochemical detection of β-naphthoflavone-and phenobarbital-induced avian cytochrome P450 enzymes

Livers from mallards ( Anas platyrhynchos ) were treated with either β-naphthoflavone (50 mg/kg) or phenobarbital (70 mg/kg). Purification of induced hepatic cytochrome P450 was accomplished using both DEAE and hydroxyapatite columns, as well as sodium dodecyl sulfate polyacrylamide gel electrophoresis separation. Polyclonal antibodies to these proteins were then produced in young male New Zealand White rabbits. β-Naphthoflavone (βNF)- and phenobarbital (PB)-treated red-winged blackbird, screech owl, European starling, and lesser scaup liver microsomes were analyzed in western blots for species cross-reactivity. Although all four of these avian species exhibited cross-reactivity with antibodies to βNF-induced mallard P450, all but the lesser scaup revealed a protein of higher molecular weight than that of the βNF-induced mallard. In addition, only the lesser scaup exhibited cross-reactivity with the anti-PB-induced mallard P450 antibodies.

Environmental Toxicology and Chemistry↗

Transboundary pollution: Persistent organochlorine pesticides in migrant birds of the Southwestern United States and Mexico

The hypothesis that migratory birds accumulate persistent organochlorine pesticides (POPs) during the winter in Latin America has been prevalent for many years, particularly since 1,1,1-trichloro-2,2–bis(p-chlorophenyl)ethane (DDT) was banned in the United States in 1972. It has been suggested that peregrine falcons ( Falco peregrinus ), black-crowned night herons ( Nycticorax nycticorax ), white-faced ibises ( Plegadis chihi ), various migratory waterfowl and shorebirds, and other avian species accumulate higher concentrations of POPs while on migration or on their wintering grounds in Latin America. Nonetheless, the data obtained thus far are limited, and there is no clear pattern to suggest that such accumulation occurs on a widespread basis. In this review wildlife contaminant studies conducted along the U.S.-Mexico border and throughout Mexico are discussed. The results for the most part seem to indicate that no major accumulation of 2,2-bis(p-chlorophenyl)-1,1-dichloroethylene) (DDE), the most persistent organochlorine compound, has occurred or been reported for most parts of Mexico. The majority of the DDE values in birds from Mexico were similar to those reported in birds from the southwestern United States during the same years. More work needs to be done, particularly in those cotton-producing areas of Mexico where DDT was applied heavily in the past (e.g., Chiapas and Michoacan). Because DDT is still used for malaria control and may still be used in agriculture in Chiapas, this state is probably the one where most migrant species would still be at a significant risk of increased accumulation of DDE and DDT.

Environmental Toxicology and Chemistry↗

Contaminant concentrations and biomarker response in great blue heron eggs from 10 colonies on the upper Mississippi River, USA

In 1993, great blue heron ( Ardea herodias ; GBH) eggs were collected from 10 colonies on the upper Mississippi River (UMR). They were then artificially incubated until pipping and analyzed for mercury, selenium, and organochlorines. Livers of embryos were analyzed for hepatic microsomal ethoxyresorufin- O -dealkylase (EROD) activity and four measures of oxidative stress. Brains were measured for asymmetry and blood was measured for the coefficient of variation of DNA (DNA CV). Organochlorine concentrations were generally low (geometric mean DDE = 1.3 μg/g wet weight; polychlorinated biphenyl [PCB] = 3.0 μg/g; 2,3,7,8-tetrachlorodibenzo- p -dioxin [TCDD] = 11.5 pg/g). Eggshell thickness was negatively correlated with DDE concentrations. Mercury (geometric mean = 0.8 μg/g dry weight) and selenium (3.1 μg/g dry weight) concentrations in GBH eggs were within background levels. EROD activity was not correlated with total PCBs, TCDD, or toxic equivalents (TEQs), based on the relative contribution of individual PCB congeners, dibenzodioxins (PCDDs), and dibenzofurans (PCDFs) to total calculated TEQs. Three of the four measures of oxidative stress were correlated with mercury concentrations. Twenty of 43 (47%) embryo brains were asymmetrical and the embryos with asymmetrical brains had higher EROD concentrations in the liver and higher DNA CV in the blood than embryos with symmetrical brains.

Environmental Toxicology and Chemistry↗

Polychlorinated dibenzo-p-dioxins, dibenzofurans, biphenyls and 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents in fishes from Saginaw Bay, Michigan

Concentrations of polychlorinated biphenyls (PCBs), polychlorinated dibenzo- p -dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), polychlorinated naphthalenes (PCNs), polychlorinated dibenzothiophenes (PCDTs), and 2,3,7,8-tetrachlo-rodibenzo- p -dioxin (2,3,7,8-TCDD) equivalents were measured in composite samples of forage fishes and several sizes of walleyes ( Stizostedion vitreum ) and common carp ( Cyprinus carpio ) collected from Saginaw Bay, Michigan, USA. The concentrations of 2,3,7,8-TCDD equivalents determined from the use of TCDD equivalency factors (TEFs) and instrumentally determined concentrations of individual congeners were designated TEq. The 2,3,7,8-TCDD equivalents determined on the same extracts with the H4IIE bioassay were defined as TCDD-EQ. Concentrations of TCDD-EQ and TEq were significantly, but poorly correlated ( R 2 = 0.44). Concentrations of TCDD-EQ and TEq ranged from 10.6 to 348 and 13.5 to 69.5 ng/kg, wet weight, respectively. The ratio of TCDD-EQ to TEq ranged from 0.5 to 5.0, with the greatest ratio observed in common carp but the average ratio between TEq and TCDD-EQ was approximately 1.0. In common carp, larger walleye, and medium-sized alewife ( Alosa pseudoharengus ) there were significant concentrations of TCDD-EQ that could not be accounted for by TEq calculated from an additive model of PCBs, PCDDs, or PCDFs. As much as 75% of the TCDD-EQ in some samples could not be accounted for by TEqs. It is unlikely that PCNs, PCDTs, or polychlorinated diphenyl ethers (PCDEs) occurred at concentrations that were, based on their relative potencies, great enough to contribute significant quantities of TCDD-EQ. The discrepency between TCDD-EQs and TEqs was not due to antagonisms among congeners or extraction efficiencies.

Michigan↗

Acute toxicity of fire-retardant and foam-suppressant chemicals to yalella azteca (Saussure)

Acute toxicity tests were conducted with Hyalella azteca Saussure (an amphipod) exposed in soft and hard waters to three fire retardants (Fire-Trol GTS-R, Fire-Trol LCG-R, and Phos-Chek D75-F) and two foam suppressants (Phos-Chek WD-881 and Silv-Ex). The chemicals were slightly to moderately toxic to amphipods. The most toxic chemical to amphipods in soft and hard water was Phos-Chek WD-881 (96-h mean lethal concentration [LC50] equal to 10 mg/L and 22 mg/L, respectively), and the least toxic chemical to amphipods in soft water was Fire-Trol GTS-R (96-h LC50 equal to 127 mg/L) and in hard water was Fire-Trol LCG-R (96-h LC50 equal to 535 mg/L). Concentrations of ammonia in tests with the three fire retardants and both water types were greater than reported LC50 values and probably were the major toxic component. Estimated un-ionized ammonia concentrations near the LC50 were frequently less than the reported LC50 ammonia concentrations for amphipods. The three fire retardants were more toxic in soft water than in hard water even though ammonia and un-ionized ammonia concentrations were higher in hard water tests than in soft water tests. The accidental entry of fire-fighting chemicals into aquatic environments could adversely affect aquatic invertebrates, thereby disrupting ecosystem function.

Environmental Toxicology and Chemistry↗

Effects of 3,3′,4,4′,5-pentachlorobiphenyl (PCB 126), 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), or an extract derived from field-collected cormorant eggs injected into double-crested cormorant (Phalacrocorax auritus) eggs

Double-crested cormorant ( Phalacrocorax auritus ) eggs were injected with either 3,3′,4,4′,5-pentachlorobiphenyl (PCB 126), 2,3,7,8-tetrachlorodibenzo- p -dioxin (TCDD), or an extract derived from field-collected double-crested cormorant eggs. These compounds were injected into the yolks of cormorant eggs from an isolated colony on Lake Winnipegosis, Manitoba, Canada. Upon hatching, chicks were necropsied. The brain, bursa, heart, liver, and spleen were removed and weighed. An approximate median lethal dose (LD50) of 158 μg/kg egg was determined for PCB 126, which is 69 times greater than the LD50 determined for the chicken ( Gallus domesticus ) in a previous study. A significantly greater mortality occurred at the highest dose of TCDD (4.0 μg/kg egg) when compared to the vehicle control. However, the mortality data did not provide sufficient information for the determination of an LD50. The cormorant egg extract did not adversely affect hatchability. No significant increases were observed in the incidence of developmental abnormalities, including pronounced edema, in any of the treatment groups, nor were there any relevant effects on body and organ weights. Based on the results from this study, the cormorant appears to be considerably less sensitive to polyhalogenated diaromatic hydrocarbons than the chicken, which has been the typical species used for egg injection studies.

Manitoba↗

An automated HPLC method for the fractionation of polychlorinated biphenyls, polychlorinated dibenzo-p-dioxins, and polychlorinated dibenzofurans in fish tissue on a porous graphitic carbon column

The Ah (aryl-hydrocarbon) hydroxylase-receptor active polychlorinated biphenyls (PCBs), polychlorinated dibenzo- p -dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) were fractionated by an automated high-performance liquid chromatography (HPLC) system using the Hypercarb™ porous graphitic carbon (PGC) column. This commercially available column was used to fractionate the di-, mono-, and non- ortho PCBs into three fractions for gas chromatography (GC)/electron capture detection analysis, and a fourth fraction containing the PCDDs/PCDFs for GC/mass spectrometry analysis. The recoveries of the PCBs ranged from 68 to 96%, and recoveries of the PCDDs/PCDFs ranged from 74 to 123%. The PGC column has the advantage of faster separations (110 min versus 446 min) and less solvent use (275 ml versus 1,100 ml) compared with automated fractionation of these compounds on activated carbon (PX-21), while still affording good separation of the classes. The PGC column may have an advantage over the pyrenyl-based HPLC method because it has a greater loading capacity (400 μg total PCBs versus 250 μg). Overall, the PGC is a standard column that provides reproducible fractionation of PCDD/PCDFs and PCBs for analytical measurement in environmental samples.

Environmental Toxicology and Chemistry↗

Distribution of organochlorine contaminants in double-crested cormorant eggs and sibling embryos

Double-crested cormorant ( Phalacrocorax auritus ) fresh eggs and sibling embryos at pipping were collected from a polychlorinated biphenyl (PCB)-contaminated colony in Green Bay, Wisconsin, USA. Egg contents were analyzed for organochlorine (OC) contaminants, including 15 arylhydrocarbon-active PCB congeners. In order to determine the significance of tissue removal on the subsequent estimate of contaminant burden, embryos were decapitated and the heads, yolk sac, liver, fecal sac (allantois), and carcass remainder were analyzed separately. The distribution of contaminant concentration in the embryos was yolk sac > liver > carcass > head > fecal sac. The distribution of contaminant mass in the embryos was yolk sac > carcass > liver > head > fecal sac. For example, mass of total PCBs (TPCB) was yolk sac = 58%, carcass = 31%, liver = 5%, head = 3%, and fecal sac = 1%. Eighteen additional OCs, including 13 PCB congeners, had distribution patterns similar to that of TPCB concentration and mass. Excluding the head of the embryo from the chemical analysis overestimated TPCB concentrations by 15% (16 vs 14 μg/g). In contrast, excluding the liver from the chemical analysis underestimated TPCB concentration by only 4% (13.5 vs 14 μg/g). Mean concentrations of OCs were not significantly different between fresh eggs and sibling embryos.

Environmental Toxicology and Chemistry↗