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Biotransformation of trenbolone acetate metabolites and estrogens derived from animal feeding operations in soils, waste storage systems, and in land applied manure has been well characterized. Yet recent data demonstrate potential for steroid transport into the environment directly from feedyard pens via runoff or airborne particulate matter. Therefore, the objective of this study was to determine steroid transformation rates in beef cattle excreta. Feces and urine were collected from steers recently treated with steroidal implants. Excreta were stored and periodically extracted over 112 d then analyzed for trenbolone acetate metabolites and estrogens by liquid chromatography mass spectrometry. Conjugated steroids were present primarily in urine, and conjugates quickly degraded to free steroid with a half-life of 0.6–1.0 d. The primary trenbolone acetate metabolite, 17α-trenbolone, had a half-life of 5.1–9.5 d. Likewise, 17α-estradiol was the predominant estrogen, with a half-life of 8.6–53 d. Secondary trenbolone metabolites formed from 17α-trenbolone biotransformation were observed at low concentrations less than 10% initial 17α-trenbolone concentrations. Estrone was the primary metabolite of 17α-estradiol and concentrations of estrone exceeded initial 17α-estradiol concentration in all sample types. These results suggest manure-borne steroids are more stable in excreta than in soil microcosms.

Metal oxide nanoparticles, such as copper oxide (CuO), are mass produced for use in a variety of products like coatings and ceramics. Acute exposure to CuO nanoparticles has caused toxicity to many aquatic organisms, yet there is no information on the effect of prolonged CuO nanomaterial exposures. This study examined effects of chronic exposure to CuO nanoparticles on Xenopus laevis growth and development. Experiments included a 14 d subchronic exposure and a 47 d chronic exposure throughout metamorphosis. The subchronic exposure caused mortality in all tested CuO concentrations, and significant growth effects occurred after exposure to 2.5 mg L−1 CuO. Chronic exposure to 0.3 mg L−1 CuO elicited significant mortality and affected the rate of metamorphosis. Exposure to lower concentrations of CuO stimulated metamorphosis and growth, indicating that low dose exposure can have hormetic effects.

Metal oxide nanomaterials have exhibited toxicity to a variety of aquatic organisms, especially microbes and invertebrates. To date, few studies have evaluated the toxicity of metal oxide nanomaterials on aquatic vertebrates. Therefore, this study examined effects of ZnO, TiO(2), Fe(2)O(3), and CuO nanomaterials (20-100 nm) on amphibians utilizing the Frog Embryo Teratogenesis Assay Xenopus (FETAX) protocol, a 96 h exposure with daily solution exchanges. Nanomaterials were dispersed in reconstituted moderately hard test medium. These exposures did not increase mortality in static renewal exposures containing up to 1,000 mg L(-1) for TiO(2), Fe(2)O(3), CuO, and ZnO, but did induce developmental abnormalities. Gastrointestinal, spinal, and other abnormalities were observed in CuO and ZnO nanomaterial exposures at concentrations as low as 3.16 mg L(-1) (ZnO). An EC(50) of 10.3 mg L(-1) ZnO was observed for total malformations. The minimum concentration to inhibit growth of tadpoles exposed to CuO or ZnO nanomaterials was 10 mg L(-1). The results indicate that select nanomaterials can negatively affect amphibians during development. Evaluation of nanomaterial exposure on vertebrate organisms are imperative to responsible production and introduction of nanomaterials in everyday products to ensure human and environmental safety.

The long-term environmental impact and potential human health hazards resulting from Hurricanes Katrina and Rita throughout much of the United States Gulf Coast, particularly in the New Orleans, Louisiana, USA area are still being assessed and realized after more than four years. Numerous government agencies and private entities have collected environmental samples from throughout New Orleans and found concentrations of contaminants exceeding human health screening values as established by the United States Environmental Protection Agency (USEPA) for air, soil, and water. To further assess risks of exposure to toxic concentrations of soil contaminants for citizens, particularly children, returning to live in New Orleans following the storms, soils collected from schoolyards prior to Hurricane Katrina and after Hurricane Rita were screened for 26 metals. Concentrations exceeding USEPA Regional Screening Levels (USEPA-RSL), total exposure, non-cancer endpoints, for residential soils for arsenic (As), iron (Fe), lead (Pb), and thallium (Tl) were detected in soil samples collected from schoolyards both prior to Hurricane Katrina and after Hurricane Rita. Approximately 43% (9/21) of schoolyard soils collected prior to Hurricane Katrina contained Pb concentrations greater than 400mgkg(-1), and samples from four schoolyards collected after Hurricane Rita contained detectable Pb concentrations, with two exceeding 1700mgkg(-1). Thallium concentrations exceeded USEPA-RSL in samples collected from five schoolyards after Hurricane Rita. Based upon these findings and the known increased susceptibility of children to the effects of Pb exposure, a more extensive assessment of the soils in schoolyards, public parks and other residential areas of New Orleans for metal contaminants is warranted.

Contamination by hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) has been identified at areas of explosive manufacturing, processing, storage, and usage. Anaerobic conversion of RDX to N-nitroso metabolites (hexahydro-1-nitroso-3,5-dinitro-1,3,5-triazine (MNX), hexahydro-1,3-dinitroso-5-nitro-1,3,5-triazine (DNX), and hexahydro-1,3,5-trinitroso-1,3,5-triazine (TNX)) has been demonstrated in the environment and in gastrointestinal tracts of mammals in vivo. Thus, potential exists for exposure to these N-nitroso compounds. While exposed to TNX via drinking water ad libitum, deer mice (Peromyscus maniculatus) were bred in three generations to produce cohorts F1A-D, F2A-B, and F3A. TNX was administered at four exposure levels: control (0 microg L(-1)), 10 microg L(-1), 100 microg L(-1), and 1000 microg L(-1). Endpoints investigated include: offspring production, offspring survival, offspring weight gain, and offspring organ weights. TNX exposure decreased litter size and increased postpartum mortality of offspring at the highest exposure level.

We evaluated the use of the gas exchange rate as an ecologically relevant indicator of chemical stress in avian embryos/eggs. Northern bobwhite quail (Colinus virginianus) were exposed to octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX) via feed containing nominal concentrations of 0, 12.5, 50.0, and 125.0 mg kg(-1). Metabolic rates (oxygen consumption) of developing quail eggs were then measured via respirometry to examine potential effects of HMX exposure. Metabolic rates were examined on 5, 9, and 21 d of incubation. Next, concentrations of HMX in embryos/eggs were determined by liquid chromatography-mass spectrometry. Mean (+/-SE) concentrations of HMX in eggs were 21.0+/-5.9, 1113+/-79.0, 3864+/-154.0, and 7426+/-301.1 ng g(-1) in control, low, medium and high dose groups, respectively. There were significant differences in oxygen consumption among the three embryo ages, however differences among the ages were not consistent among dose groups (age x dose group interaction p

Given the potent carcinogenic effects of most N-nitroso compounds, the reductive transformation of the common explosive hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) to a group of N-nitroso derivatives, hexahydro-1-nitroso-3,5-dinitro-1,3,5-triazine (MNX), hexahydro-1,3-dinitroso-5-nitro-1,3,5-triazine (DNX), and hexahydro-1,3,5-trinitroso-1,3,5-triazine (TNX) in the environment have caused concerns among the general public. Questions are arising about whether the same transformations also occur in mammals, and if true, to what extent. This study investigated the N-nitroso derivatives production in the deer mouse GI tract following RDX administration. Findings verified that such transformations do occur in the mammalian GI tract at notable levels: the average MNX concentrations in deer mice stomach were 85 microg/kg and 1318 microg/kg for exposure to 10mg/kg and 100mg/kg diet, respectively. DNX in stomach were 217 microg/kg for the 10mg/kg dose group and 498 microg/kg for the 100mg/kg dose group. Changes in other toxic endpoints including body weight gain, food consumption, organ weight, and behavior were also reported.

Hexahydro-1,3,5-trinitro-1,3,5-triazine, a cyclonitramine commonly known as RDX, is used in the production of military munitions. Contamination of soil, sediment, and ground and surface waters with RDX has been reported in different places around the world. Acute and subacute toxicities of RDX have been relatively well documented in terrestrial vertebrates, but among aquatic vertebrates the information available is limited. The objective of this study was to characterize the acute toxicity of RDX to larval zebrafish. Mortality (LC50) and incidence of vertebral column deformities (EC50) were two of the end points measured in this study. The 96-h LC50 was estimated at 22.98 and 25.64 mg l −1 in two different tests. The estimated no-observed-effective-concentration (NOEC) values of RDX on lethality were 13.27 ± 0.05 and 15.32 ± 0.30 mg l −1 ; and the lowest-observed-effective-concentration (LOEC) values were 16.52 ± 0.05 and 19.09 ± 0.23 mg l −1 in these two tests, respectively. The 96-h EC50 for vertebral deformities on survivors from one of the acute lethality tests was estimated at 20.84 mg l −1 , with NOEC and LOEC of 9.75 ± 0.34 and 12.84 ± 0.34 mg l −1 , respectively. Behavioral aberrations were also noted in this acute toxicity study, including the occurrence of whirling movement and lethargic behavior. The acute effects of RDX on survival, incidence of deformities, and behavior of larval zebrafish occurred at the high end of the most frequently reported concentrations of RDX in aquatic environments. The chronic effects of RDX in aquatic vertebrates need to be determined for an adequate assessment of the ecological risk of environmental RDX.

Since previous studies had determined organic contamination in Savannah Harbor sediments, an assessment was needed to determine its potential effects on wildlife species which frequently used surrounding areas as feeding and breeding habitats. Ninety percent of the black neck stilt egg contents evaluated contained p,p′-DDE, and 14% of the egg contents contained more than one PCB congener. Ninety percent of chorioallantoic membranes evaluated contained p,p′-DDE. Data from this study will provide information necessary to evaluate proposed measures to enhance bird use of confined disposal facilities associated with Savannah Harbor and will serve as a baseline for future ecotoxicological information gathered to better manage bird nesting areas along the Atlantic coast.

Concentrations of mercury in biological samples collected for environmental studies are often less than 0.1 microgram/g. Low mercury concentrations and small organ sizes in many wildlife species (approximately 0.1 g) increase the difficulty of mercury determination at environmentally relevant concentrations. We have developed a digestion technique to extract mercury from small (0.1 g), biological samples at these relevant concentrations. Mean recoveries (+/- standard error) from validation trials of mercury fortified tissue samples using cold vapor atomic absorption spectroscopy for analysis ranged from 102 +/- 4.3% (2.5 micrograms/L, n = 15) to 108 +/- 1.4% (25 micrograms/L, n = 15). Recoveries of inorganic mercury were 99 +/- 5 (n = 19) for quality assurance samples analyzed during environmental evaluations conducted during a 24 month period. This technique can be used to determine total mercury concentrations of 60 ng Hg/g sample. Samples can be analyzed in standard laboratories in a short time, at minimal cost. The technique is versatile and can be used to determine mercury concentrations in several different matrices, limiting the time and expense of method development and validation.

PCBs were found in eggs and chorioallantoic membranes (CAMs) of loggerhead sea turtles (Caretta caretta). Total PCB concentrations in CAMs were larger than concentrations in eggs. Total PCB concentrations in egg and CAM tissues were highly correlated (r2 = 0.782; p = 0.0001). Sums of PCB congeners within homologue groups were also correlated in the two tissues. Data from these turtle eggs indicate 1) PCB concentrations in CAMs represent PCB concentrations in whole eggs and 2) CAMs can be collected in a nonlethal manner to determine PCB concentrations in sea turtle hatchlings.

Nitrous acid (HNO2), nitric acid (HNO3), nitric oxide (NO) and nitrogen dioxide (NO2) were measured over a four day period in Clairmont, CA. Concentrations of each nitrogen oxide (NOx) compound varied diurnally during the study period HNO3, NO2, and NO generally reached maxima during daylight hours. Conversely, HNO2 concentrations were highest at night. HNO3 and HNO2 concentrations increased with increasing NO and NO2. HNO3 dependance became limited at high NO2 and NO. HNO2 concentrations declined when high NO and NO2 concentrations were present. The decline in HNO2 is postulated to result from direct photolysis during the time periods of NO and NO2 maxima or from reactions to form HNO3.

Chorio-allantoic membranes (CAMS) were collected and analyzed for chlorinated hydrocarbons as part of a wildlife toxicology demonstration project at Naval Air Station (NAS) Whidbey Island, Washington, USA. Concentrations of DDT, DDE, DDD, Aroclor 1254, and Aroclor 1260 were found at concentrations below 0.4 ppm for 13 of 14 samples. The low correlations among DDT and its metabolites in CAMS suggest herons are not being exposed to a consistent source of these compounds. Comparison of chlorinated hydrocarbon data for CAMS from three Puget Sound heron colonies, NAS Whidbey, Samish Island and Dumas Bay, indicates contaminant burdens in herons from NAS Whidbey and Samish Island are significantly lower than burdens in herons from Dumas Bay.

Metal oxide nanomaterials have exhibited toxicity to a variety of aquatic organisms, especially microbes and invertebrates. To date, few studies have evaluated the toxicity of metal oxide nanomaterials on aquatic vertebrates. Therefore, this study examined effects of ZnO, TiO(2), Fe(2)O(3), and CuO nanomaterials (20-100 nm) on amphibians utilizing the Frog Embryo Teratogenesis Assay Xenopus (FETAX) protocol, a 96 h exposure with daily solution exchanges. Nanomaterials were dispersed in reconstituted moderately hard test medium. These exposures did not increase mortality in static renewal exposures containing up to 1,000 mg L(-1) for TiO(2), Fe(2)O(3), CuO, and ZnO, but did induce developmental abnormalities. Gastrointestinal, spinal, and other abnormalities were observed in CuO and ZnO nanomaterial exposures at concentrations as low as 3.16 mg L(-1) (ZnO). An EC(50) of 10.3 mg L(-1) ZnO was observed for total malformations. The minimum concentration to inhibit growth of tadpoles exposed to CuO or ZnO nanomaterials was 10 mg L(-1). The results indicate that select nanomaterials can negatively affect amphibians during development. Evaluation of nanomaterial exposure on vertebrate organisms are imperative to responsible production and introduction of nanomaterials in everyday products to ensure human and environmental safety.

Uptake and accumulation kinetics of hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) and its two major N-nitroso metabolites, hexahydro-1-nitroso-3,5-dinitro-1,3,5-triazine (MNX) and hexahydro-1,3,5-trinitroso-1,3,5-triazine (TNX), in earthworms was investigated. Results indicated that RDX and its N-nitroso metabolites were rapidly absorbed into earthworms (Eisenia fetida), reaching the highest concentrations within a few days. Accumulation of RDX was greater than its N-nitroso metabolites, as evidenced by a higher bioconcentration factor (BCF); BCFs were 1.86, 0.39, and 0.05 for RDX, MNX, and TNX, respectively. RDX and its N-nitroso metabolites were also rapidly eliminated from the earthworm and/or transformed to other metabolites, as evidenced by the rapid decrease of test compounds in earthworms after reaching their highest concentrations. The uptake of MNX and TNX increased as exposure concentration increased. Although these earthworms might (anaerobically) degrade RDX to MNX and MNX to TNX, it is hypothesized that this process would be slow. Other biotransformation pathways may be involved in biodegradation of RDX and its N-nitroso metabolites due to the fact that concentrations of tested compounds decreased in both soil and earthworms. It is hoped that these data can be used to refine environmental management strategies for RDX and for performing specific risk assessments of RDX and its N-nitroso metabolites.

Hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) transforms anaerobically into N-nitroso compounds: hexahydro-1-nitroso-3,5-dinitro-1,3,5-triazine (MNX), hexahydro-1,3-dinitroso-5-nitro-1,3,5-triazine (DNX), and hexahydro-1,3,5-trinitroso-1,3,5-triazine (TNX). Exposure to these N-nitroso metabolites may occur in areas contaminated with explosives, as anaerobic degradation occurs via some bacteria and is one remediation strategy used for RDX. Few papers report acute oral toxicity and none have evaluated age dependent toxicity of RDX or its N-nitroso metabolites. Median lethal dose (LD50) was determined in deer mice (Peromyscus maniculatus) of three age classifications 21 d, 50 d, and 200 d for RDX, MNX, and TNX using the US EPA up-and-down procedure (UDP). Hexahydro-1,3,5-trinitro-1,3,5-triazine and N-nitroso metabolites caused similar overt signs of toxicity. Median lethal dose for 21 d deer mice were 136, 181, and 338 mg/kg for RDX, MNX, and TNX, respectively. Median lethal dose for 50 d deer mice were 319, 575, and 338 mg/kg for RDX, MNX, and TNX, respectively. Median lethal dose for 200 d deer mice were 158, 542, and 999 mg/kg for RDX, MNX, and TNX, respectively. These data suggest that RDX is the most potent compound tested, and age dependent toxicity may exist for all compounds and could play a role in RDX and RDX N-nitroso metabolite ecological risk evaluation of terrestrial wildlife at RDX contaminated sites.

In toxicokinetics studies, interactions between chemicals in mixtures has been largely neglected. This study examines a mixture of perchlorate and arsenate because (1) they have the potential to co-occur in contaminated aquatic habitats, and (2) a previous study by the authors found possible toxicological interactive effects. In the present study, zebrafish (Danio rerio) were exposed to two concentrations of sodium perchlorate (10 and 100 mg l−1), sodium arsenate (1 and 10 mg l−1), and the mixture-sodium perchlorate + sodium arsenate (10 + 1 mg l−1 and 100 + 10 mg l−1 Na2HAsO4-high mixture) for 90 d. Their uptake and accumulation by zebrafish was evaluated at 10, 30, 60, and 90 d. In addition, depuration was examined at 1, 3, and 5 d after cessation of the exposure. The uptake of either chemical was concentration-dependent, with significantly higher uptake at high concentrations at either exposure interval. In contrast, there was no significant difference in whole body residue between single chemicals and the corresponding mixture except for 100 mg l−1 sodium arsenate at 90 d. However, there was increasing accumulation over time at the high concentration of either chemical alone and their mixture, and this increasing trend was more pronounced in the single chemical exposures than in the mixture. At the concentrations tested in the current study, both chemicals reduced the uptake but enhanced the depuration of the other chemical from the zebrafish. This study represents the first examination of the interaction of two anions-perchlorate and arsenate with respect to toxicokinetics.

Livers of bullfrogs (Rana catesbeiana) from a polychlorinated biphenyl (PCB) contaminated watershed and hazardous waste site located in Pickens County, South Carolina, contained significantly higher concentrations of PCBs (2.33 and 2.26 ppm, respectively) than those from a reference site (0.05 ppm). Green frogs (R. clamitans) from the two contaminated sites also accumulated higher levels of PCBs (2.37 and 3.88 ppm, respectively) than those from the reference site (0.02 ppm). No temporal variation was observed in PCB concentrations of bullfrogs or green frogs from the contaminated sites between 1992 and 1993. Levels of PCBs in the livers of northern water snakes (Nerodia sipedon) were significantly higher in snakes from the contaminated watershed (13.70 ppm) than in those from the waste site (2.29 ppm) and two reference sites (2.50 and 1.23 ppm). When compared to frogs, significantly higher bioaccumulation occurred in water snakes from the contaminated watershed. No significant differences in PCB levels were found with respect to sex or body size (snout-vent length (SVL) or body mass) for frogs or snakes. PCBs were detected also in eggs of both frogs and snakes. Results of this study provide baseline data and document the bioaccumulation of PCB residues in frog and snake tissues; however, the significance of these tissue residues to reproduction, survival, growth/development, and population dynamics in contaminated habitats is unknown.