RP084 (LIM-1117-818754) Analysis of TCDD-Induced Oxidative Stress in Chicken Liver During Development.
Start time: 8:00 AM
Lim, J¹, DeWitt, J.C², Watkins III, J.B¹, Henshel, D.S¹, ¹ Indiana University, Bloomington, IN, U.S.A² US EPA NHEERL/ETD/ITB, Research Triangle Park, NC, U.S.A
2,3,7,8-tetrachlorodibenzo-p- dioxin (TCDD) is known to induce oxidative stress during development in animals. TCDD exposure during development does not lead to direct genetic alterations, but TCDD-induced oxidative stress may play a major role as a promoter and/or an initiator of malignant tumors. Thus, exposure to TCDD is believed to increase radical oxygen species (ROS) production mainly in liver and affect major endogenous antioxidants associated with removing ROS in cellular components. This research focused on determining the activity changes of antioxidant enzymes such as glutathione peroxidase (GPx), glutathione reductase (GRx), catalase, and superoxide dismutase (SOD) in chicken liver during development. TCDD of 2 ppt, 20 ppt, and 200 ppt was injected into chicken eggs via the airsac at embryonic day zero (E0) with a no-injection control (NIC) and a vehicle control (sunflower oil) to induce oxidative stress in hatchling chicks. Liver tissue (0.3g) was homogenized and changes in activity of the antioxidant enzymes were determined to evaluate the levels of oxidative stress. Reduction of enzyme activity in GPx, GRx, catalase, and SOD was apparent; a 45∼67% decrease in GPx, a 50∼54% in GRx, a 20∼32% in catalase, a 35%∼46% in SOD. No statistically significant differences were observed between TCDD dose groups (P-values >0.05), which shows that any does-response relationship was not recognized. However, this study demonstrates that the induction of oxidative stress by embryonic TCDD exposure in ovo leads to the significant decrease of the antioxidant enzyme activity and thus is likely to contribute to the embryotoxicity of TCDD in the hatchling chicks.

RP085 (MIL-1117-837037) Heart ventricle wall thinning in five species of passerines exposed to environmental mixtures of PCBs during development.
Start time: 8:00 AM
Millsap, D¹, Wakhungu, P², Heise, S³, Yeager, R⁴, Sparks, D⁵, Henshel, D⁶, ¹ Indiana University, Bloomington, IN, USA² Indiana University, Bloomington, IN, USA³ Indiana University, Bloomington, IN, USA⁴ Indiana University, Bloomington, IN, USA⁵ U.S. Fish and Wildlife Service, Bloomington, IN, USA⁶ Indiana University, Bloomington, IN, USA
Five passerine species collected from PCB-contaminated sites and a reference site were quantitatively assessed for external heart morphometrics and deformities. Nestling hearts were transversely sectioned, digitally photographed, and ventricle wall thickness measurements were taken using Image Pro Plus 5.0 software. External heart morphometrics and deformities were correlated with left, right, and inner septa ventricle wall measurements for each species and site. Tree swallow nestlings from PCB-contaminated sites showed a statistically significant increase in right ventricle wall thinness as compared to the reference site. There was also a statistically significant correlation between the external deformities center roll, macrosurface roughness, and ventricular notching in tree swallow nestlings from PCB-contaminated sites as compared to the reference site. No statistically significant correlations were found between external deformities and ventricle wall thinness for Carolina chickadee, eastern bluebird, house wren, or red-winged blackbird.

RP086 (YES-1117-805201) Evaluation of the genotoxicity of some textile dyes and wastewater.
Start time: 8:00 AM
Yesilada, E¹, Ozata, L², Dogan, E³, ¹ Inonu University, Malatya, Turkey² Inonu University, Malatya, Turkey³ Inonu University, Malatya, Turkey
In the present study, four textile dyes namely Chrocion Scarlet, Chrocion Yellow, Remazol Turquoise Blue, Remazol Red RR and textile wastewater were evaluated for mutagenic and recombinagenic effects using the wing spot test of Drosophila melanogaster (somatic mutation and recombination test, SMART). The standard cross (ST) and the improved high-bioactivation cross (HB) were used. The latter cross is characterized by a high sensitivity to promutagens and procarcinogens. Three-day-old larvae, transheterozygous for the multiple wing hairs and flare genes, were chronically fed with different concentrations of test compounds. Feeding ended with pupation of the surviving larvae and the genetic changes induced in somatic cells of the imaginal discs of the wings lead to the formation of mutant clones on the wing blade. Point mutation, chromosome breakage and mitotic recombination produce single spots; while twin spots are produced only by mitotic recombination. All concentrations of textile dyes and textile wastewater, ethyl methanesulfonate (EMS) and urethane caused a decrease in survival proportional to concentration used. Treatment of the standard and the high-bioactivation crosses with Chrocion Yellow, Remazol Turquoise Blue and Remazol Red RR gave positive results, apparent from increase in the frequency of the single spots. Chrocion Scarlet gave inconsistent result in both crosses. Textile wastewater increased the number of all spots type in both crosses and proved to be more genotoxic in the HB cross than in the ST cross. This study proved the genotoxicity of these textile dyes, and suggested that further studies should be made on other dyes and some other toxic industrial pollutant discharges in ecosystems, using D. melanogaster wing spot test as an indicator to monitor pollutant genotoxicity.

RP087 (HAM-1117-846640) Comparative Analysis of the Toxic Effects of Perchlorate in Arabidopsis and in Sorghum.
Start time: 8:00 AM
Hamissou, Mijitaba¹, Dunkerley, Ray¹, ¹ Jacksonville State University, Jacksonville, Alabama, USA
Perchlorate contamination in ground water was recently reported in several southwestern states of the United States. It is used in rocket fuel, airbags, matches, and ammunition. When released into the environment, perchlorate becomes an environmental pollutant posing potential immunotoxic conditions in plants and animals. Environmental contaminants alter plant growth and metabolism. However, some plant species have evolved to tolerate or accumulate soil and water contaminants by extracting them from their immediate surrounding. The toxic actions of perchlorate are believed to be exerted on key metabolic enzymes. Like heavy metal pollutants, perchlorate may induce a burst of reactive oxygen species (ROS) creating therefore conditions for secondary oxidative stress to the plants. Literatures reviewed have indicated that higher plants respond to environmental pollutants by sequestering them in their vacuoles or by synthesizing phytochelatin and small molecular weight proteins to neutralize the toxic effects. In this research, mortality, growth parameters, and the activities of ROS scavenger molecules, superoxide dismutase (SOD) and peroxidase (PO) are investigated in arabidospsi thaliana and sorghum bicolor grown in perchlorate solutions. Plants were screened in potted soil and hydroponically in perchlorate containing solutions, adjusted to pH 6.9 for 14 days. Chlorosis, biomass production, and mortality rate were measured daily for 14 days. SOD and PO activities were determined from the cytoplasmic proteins and isolated chloroplasts at the conclusion of the experiment. The data indicated that perchlorate affect arabidopsis more severely than sorghum. The oxidative responses, as measured by the activities of SOD and PO, were significantly higher in Arabidopsis than in sorghum in perchlorate treated plants.

RP088 (SMI-1117-645717) Influence of seed type on risk of exposure to granivorous birds foraging on pesticide-treated seeds.
Start time: 8:00 AM
Mineau, P¹, Smith, G², ¹ Canadian Wildlife Service, Ottawa, Ontario, Canada² Carleton University, Ottawa, Ontario, Canada
The pre-sowing application of pesticides to seeds is a very popular and potentially efficient method of crop protection. However, these seed treatments can pose a serious threat to the health of many agriculturally associated granivorous birds, sometimes having lethal effects. Assessing the potential risk of exposure from a seed treatment involves the weighted incorporation of several factors (e.g., density of exposed seeds, bird hunger, presence of other foods, and social feeding effects). An additional factor, and one that is commonly under emphasized, is the attractiveness of the crop seed in question to local birds. In North America, there is a shortage of information on the species of birds foraging in agricultural areas, and their relative preference for various crop seeds. Accordingly, we conducted a field study to identify bird species that are at risk to exposure from seed treatments by assessing their foraging habits on five locally grown types of crop seed (corn, barley, oat, wheat, soybean). Feeding stations were positioned at the edge of 10 actively farmed agricultural fields around Ottawa, Ontario, Canada, in summer/fall of 2004 and spring 2005. Video cameras were placed at each station at early dawn and data on species, crop seed attractiveness, meal size, and foraging behaviour assessed at a later date from the recordings. These results were incorporated with established toxicological data for specific seed-treatment pesticides and used to calculate theoretical exposure levels to foraging granivorous birds. These data will provide assistance in making more accurate risk assessments for bird species feeding on pesticide-treated seed.

RP089 (QUI-1116-004984) Developmental and behavioral effects on reproduction from androgen disruption in Japanese quail.
Start time: 8:00 AM
Quinn, Jr., M¹, Lavoie, E², Thompson, N², Touart, L³, Ottinger, M, ¹ U.S. Army Center for Health Promotion and Preventive Medicine, Aberdeen Proving Ground, MD, USA² University of Maryland, Animal and Avian Sciences, College Park, MD, USA³ US Environmental Protection Agency, Washington, DC, USA
In avian species, gonadal differentiation occurs during early embryonic development. A sexually dimorphic pattern of gonadal steroids that are produced during this time helps to modulate the sexual differentiation of accessory sex structures and neuroendocrine systems that regulate endocrine and behavioral components of reproduction. This, as well as differentiation of the hypothalamic-pituitary-gonadal axis, occurs mainly during late embryonic and early posthatch development. This critical period later determines not only male or female endocrine patterns, but sexually dimorphic behavioral patterns as well. Embryonic exposure to androgen-active chemicals can, therefore, cause lasting alterations in adult reproductive function. We exposed quail embryos to either 0.5, 5, 50, or 125 g trenbolone acetate, an androgenic chemical, at day 4 of incubation, or 20.0 or 40.0 g p,p-DDE [ethylene, 1, 1-dichloro-2,2-bis(p-chlorophenyl)], an anti-androgen, at day 1 of incubation. Developmental endpoints included onset of puberty, ovarian follicle counts, male proctodeal gland size, testicular weight, sperm motility, and sperm penetration of the germinal disc. Behavioral endpoints measured included latency to mount, latency to successful copulation, and number of successful cloacal contacts. Onset of puberty was delayed for over a month in males exposed to trenbolone and accelerated by approximately 8 days in females exposed to DDE. Male copulatory behavior was depressed in birds exposed to both chemicals, as measured by the number of attempts to mount and latency to mount and successfully copulate. This study illustrates that although exposure levels of endocrine disrupting chemicals may not always cause outright toxicity in wild populations, more subtle and non-lethal effects, that may otherwise go undetected in field studies, are able to elicit long term alterations in development that can cause detrimental effects on a population′s fitness.

RP090 (QUI-1116-004201) Embryonic estradiol exposure disrupts avian immune system development in Japanese quail.
Start time: 8:00 AM
Quinn, Jr., M¹, McKernan, M², Lavoie, E³, Hoffman, B³, Touart, L⁴, Ottinger, M³, ¹ .S. Army Center for Health Promotion and Preventive Medicine, Aberdeen Proving Ground, MD, USA² USGS, Patuxent Wildlife Research Center, Laurel, MD, USA³ University of Maryland, Animal and Avian Sciences Department, College Park, MD, USA⁴ US Environmental Protection Agency, Washington, DC, USA
An embryo dose range finding protocol was developed to address dose range needs for the avian two generation test. This study evaluated sensitivity of immune endpoints in Japanese quail embryonically exposed to estradiol. Eggs were injected with vehicle control (sesame oil) or 0.5, 50 or 500 g/egg of estradiol (E2) in sesame oil at embryonic day 4. Quail were sampled on day of hatch and at maturity. Bursa of Fabricius and spleen weights were measured in hatchlings and adults. Bursal morphology was assessed in hatchlings, and bursal follicle size and numbers were measured. Bursa weights were decreased in a dose dependent, however adult bursas from the 500 g treatment group were larger than controls. Spleen mass remained unchanged in both hatchlings and adults. Morphological abnormalities in the bursa included thickening of the epithelial layers surrounding the bursal plicae that were more than three times as thick as controls. Although follicle number remained unchanged, follicle size was reduced in the 50 and 500 g hatchling treatment groups. This study identifies the bursa of Fabricius as a sensitive target of estradiol. The main role of the bursa is B cell maturation, therefore development of overall immunocompetence is dependent on the maturation of this lymphoid organ. The effects of estradiol on the bursa support the potential for disruption of immune system development by embryonic exposure to estrogenic endocrine disrupting chemicals. Funded by Battelle Labs in support of a U.S. EPA contract.

RP091 (YEA-1117-827341) Chicken embryonic survivorship as a function of phototherapy-dioxin co-exposure.
Start time: 8:00 AM
Yeager, R¹, Millsap, D ¹, Lim, J¹, Heise, S¹, Eells, J², Whelan, H³, Henshel, D¹, ¹ Indiana University, Bloomington, IN, USA² University of Wisconsin-Milwaukee, Milwaukee, WI, USA³ Medical College of Wisconsin, Milwaukee, WI, USA
Photobiomodulation by red to near-infrared (630-1000 nm) light has been shown to stimulate mitochondrial energy (ATP) production, accelerate wound healing and promote cellular survival following a toxic insult. 2,3,7,8 Tetrachlorodibenzo-p-dioxin (TCDD) increases late embryo mortality in chickens by causing biochemical changes that culminate in decreased available energy. We tested the hypothesis that in ovo treatment with a red light-emitting diode (LED) array would attenuate TCDD-related embryo mortality. Domestic chicken eggs (Gallus gallus) were divided into ten treatment groups (no-inject, sunflower oil vehicle, and 2, 20, 200 ppt TCDD), all with or without LED treatment (670 nm at a fluence of 4 J/cm² at 24 hr. intervals). We observed an LED-induced decrease in mortality in the 200 ppt dioxin treated group (41% decrease in mortality rate). When week-three mortality was analyzed by correcting for the death occurring within the first two weeks of incubation, there remained a 23% decrease in mortality for the LED versus non-light treated chicks exposed to 200ppt dioxin. Thus, at doses of TCDD in excess of the 100ppt lethality threshold, LED treatment significantly attenuates TCDD toxicity. In contrast, at sub-lethal doses of TCDD (2 and 20ppt) and in oil-injected vehicle controls, LED-treatment increased the mortality rate. Thus an oil-NIR light interaction appears to contribute to early death at sub-lethal doses of TCDD. Future studies in our laboratory are aimed toward quantifying oxidative stress, P450 induction, and ATP levels in order to elucidate the mechanisms behind these relationships, as well as understanding the oil-light interaction effects.

RP092 (AUG-1117-821209) Embryotoxicity of 2,3,7,8-TCDD to the wood duck (Aix sponsa).
Start time: 8:00 AM
Augspurger, T¹, Di Giulio, R², Tillitt, D³, ¹ U.S. Fish and Wildlife Service, Raleigh, NC, USA² Duke University, Durham, NC, USA³ U.S. Geological Survey, Columbia, MO, USA
Eggs of wood ducks (Aix sponsa) collected from uncontaminated sites were injected with 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) to determine the wood duck's sensitivity relative to other birds. Four separate groups of 66 to 97 eggs (split into six treatments) were injected in 2004 and 2005. TCDD was dissolved in triolein which was injected at 0.1 microliters per gram egg. Injections were made into the yolk at embryonic day 0 and eggs were incubated with weekly assessments of mortality. Treatments included doses from 0 to 4,600 pg TCDD /g egg. Statistically significant embryo mortality was not observed at these concentrations through incubation day 25 (representing 90% of the incubation period for this species). Among 13 other avian genera for which egg-injected TCDD or polychlorinated biphenyl congener embryo lethality data are available (from the literature), our 4,600 pg/g No Observed Adverse Effect Level (NOAEL) ranks near the middle. Because this NOAEL for embryo mortality is unbounded (no higher concentrations were assessed), the wood duck appears to be among the more TCDD-tolerant of the species that have been assessed by egg injection. There was about a 9-fold increase in hepatic EROD activity over controls in the 4,600 pg/g treatment. Significant hepatic P4501A induction was observed at doses of 1,500 pg/g and 4,600 pg/g based on immunohistochemical staining. When exposures were extended to 2-weeks post-hatch (to allow total resorption of yolk and therefore exposure to all yolk-associated TCDD), there was still no statistically significant embryo mortality (doses from 0 to 3,200 pg/g). Liver, heart, eye and brain histology and hepatic EROD measurements from the longer exposures are pending.

RP093 (WIL-1117-821122) Development of an extraction method and in vitro bioassay for the putative AVM toxin.
Start time: 8:00 AM
Wiley, F^{1, 2}, Twiner, M², Van Dolah, F², Bowerman, W¹, Wilde, S³, Leighfield, T², ¹ Clemson University, Pendleton, SC, USA² NOAA-National Ocean Service, Charleston, SC, USA³ South Carolina Department of Natural Resources, Charleston, SC, USA
Avian vacuolar myelinopathy (AVM), a neurological disease of unknown origin, has been linked to ingestion of nuisance aquatic vegetation. The pathology and nature of AVM epornitics suggest a toxin as the causative agent, and the working hypothesis is that a species of epiphytic cyanobacteria associated with the vegetation is the source of this putative toxin. The purpose of this project is to develop a method for putative toxin extraction, as well as develop a rapid and sensitive in vitro assay capable of toxin detection, which can then be used to guide toxin fractionation and isolation studies. Samples of aquatic vegetation (Hydrilla verticillata and associated epiphytes) were collected and confirmed as either positive or negative for the putative AVM toxin by mallard (Anas platyrhynchos) bioassay. Equivalent amounts of positive and negative (control) vegetation are now being processed using a solvent extraction series of increasing polarity (hexane, acetone, methanol, water). Each resulting fraction will be tested for the presence of AVM toxin by mallard bioassay, as well as screened for cytotoxicity using several cell lines. Preliminary cell line results reveal cytotoxicity associated with the non-polar hexane fraction of the AVM-positive vegetation, as well as the acetone and methanol fractions of both AVM-positive and AVM-negative vegetation. The extracts require confirmation by mallard bioassay before hypothesizing a link between cytotoxicity and a putative AVM toxin.

RP094 (STO-1117-748705) Genotoxic effects of in ovo herbicide exposure in domestic chickens and ducks.
Start time: 8:00 AM
Stoddart, R.¹, Wayland, M.², Wolf, T.³, Wickstrom, M.¹, ¹ University of Saskatchewan, Saskatoon, Saskatchewan, Canada² Environment Canada, Saskatoon, Saskatchewan, Canada³ Agriculture and Agri-Foods Canada, Saskatoon, Saskatchewan, Canada
Agricultural practices such as low tillage and fall seeding, which are intended to conserve water and topsoil, often result in increased ground cover in the early spring. In prairie habitat, this sparse cover provides preferred nesting sites for waterfowl and upland game birds. Because the nesting period often coincides with herbicide treatment of many crops, eggs of ground nesting birds have the potential to be exposed during spraying. Common herbicide formulations used for weed control on the Canadian prairies include 2,4-dichlorophenoxy acetic acid (2,4-D) and Buctril-M® (bromoxynil:MCPA). There is concern about potential sublethal effects of these herbicides on developing birds. This study assessed the effects of in ovo exposure to 2,4-D ester or Buctril-M® herbicide formulations on DNA integrity in newly hatched domestic chickens (Gallus gallus) and ducks (Anas platyrhynchos). Eggs (20 per treatment group) were sprayed at either normal field application rates or at 10 times recommended rates, on days 6 or 15 (chickens) and 6 or 21 (ducks) of incubation to evaluate risks from herbicide exposure during early or late developmental stages, respectively. Control groups consisted of eggs sprayed with water only, and eggs injected with cyclophosphamide, a known genotoxicant. Potential damage to genetic material was evaluated using flow cytometry to measure DNA variability in red blood cells collected from 21-day-old birds. This method estimates the variation in DNA content among cells, which increases as a result of abnormal distribution caused by chromosomal damage. DNA content differences within individuals are calculated as the half peak coefficient of variation (HPCV). HPCV values were compared using a nested ANOVA model to detect differences among treatment groups (p> 0.05).

RP095 (HOF-1117-676968) Phosphorus amendment reduces hematological, hepatic and renal toxicity of lead-contaminated sediment to mallards.
Start time: 8:00 AM
Hoffman, D.¹, Heinz, G.¹, Audet, D.², ¹ USGS Patuxent Wildlife Research Center, Laurel, Maryland, USA² U.S. Fish and Wildlife Service, Spokane, Washington, USA
Ingestion of lead-contaminated sediments has resulted in lead poisoning of waterfowl for decades in the Coeur d Alene River Basin in Idaho. This study examined whether the addition of phosphoric acid to contaminated sediments would reduce the bioavailability and toxicity of lead to mallards (Anas platyrhynchos). Mallards received diets containing 12% clean sediment (controls) or 12% sediment from each of three different sites containing up to 6990 ug /g lead (dw) with or without phosphoric acid amendment for 8 weeks. Amendment resulted in reductions in tissue lead concentrations of up to 64% for blood, 57% for liver, and 77% for kidney. Unamended lead-contaminated sediment resulted in the following hematological and plasma alterations: 90% or greater depression of red blood cell ALAD activity, elevated free erythrocyte protoporphyrin (FEP) concentration, lower hematocrit and hemoblobin concentrations (as much as 30%), elevated plasma enzyme activities (ALT, CK and LDH-L) and creatinine concentration. Hepatic effects included: 1.6 fold elevation of liver GSH concentration, higher GSH S-transferase and GSSG reductase activities, and lower PBSH concentration. Renal effects included 2.1 fold elevation of kidney GSH concentration with resulting lower GSSG to GSH ratios, elevated GGT activity, and 1.7 fold increase in lipid peroxidation (TBARs). Phosphorus amendment restored hematocrit, hemoblobin and plasma enzyme activities so that they did not differ from controls and lowered elevated FEP concentrations by up to 80%. Amendment restored all hepatic variables as well as the renal variables TBARS concentration and GGT activity so they did not differ from controls. Although amendments of phosphorus substantially reduced the bioavailability of lead and some of the toxic effects, lead concentrations in the tissues of mallards fed the amended sediments were still above those believed to be harmful to waterfowl under the present conditions.

RP097 (NEW-1117-721057) Avian toxicity reference values (TRVs) for perfluorooctane sulfonate (PFOS).
Start time: 8:00 AM
Newsted, J¹, Jones, P², Coady, K³, Giesy, J^{2, 4}, ¹ Entrix, Inc, Okemos, MI, USA² National Food Safety and Toxicology Center, Michigan State University, East Lansing, MI, USA³ Warner Southern College, Lake Wales, FL, USA⁴ Zoology Department, NFSTC, Michigan State University, East Lansing, MI, USA
Toxicity Reference Values (TRVs) and Predicted No Effect Concentrations (PNECs) were derived for perfluorooctane sulfonate (PFOS) based on the characteristics of a top avian predator. The benchmarks are protective of avian populations and were based on acute and chronic dietary exposures of northern bobwhite quail (Colinus virginianus) and mallard (Anas platyrhynchos). Toxicological endpoints included mortality, growth, feed consumption, and histopathology. Reproductive endpoints included egg production, fertility, hatchability and survival and growth of offspring. Based on USEPA Great Lakes Initiative (GLI) methodology, an uncertainty factor (UF) of 36 was derived. Using this UF, the TRV based on PFOS dietary intake was 0.021 mg PFOS/kg/d while for serum and liver TRVs were 1.7 ug PFOS/ml and 0.6 ug PFOS/g ww, respectively. Based on European Comission (EC) methodology, a correction factor of 2 (for LOEL to NOEL) and an assessment factor of 30, for a total adjustment of 60, were used to derive PNECs. PNECS based on dietary and mean serum and liver PFOS concentrations were 0.013 mg PFOS/kg, bw/d, 1.0 ug PFOS/ml and 0.35 ug PFOS/g ww, respectively. Due to the conservative assumptions used the analyses, PFOS concentrations at or less than these values would not be expected to pose risks to avian populations. However, in light of laboratory no-observed-effect levels, population-level effects would not actually be expected to occur until 6.0 mg PFOS/kg, diet, 4.0 ug PFOS/g ww liver or 8.0 ug PFOS/ml serum, was exceeded.

RP098 (DAI-1117-833424) Liver Metal Concentrations in Greater Sage-grouse.
Start time: 8:00 AM
Dailey, R.¹, Raisbeck, M.¹, Siemion, R.¹, Cornish, T.¹, ¹ Wyoming State Veterinary Laboratory, Laramie, WY, US
Metal reference ranges for any species are valuable diagnostic tools. Reference ranges can help identify a variety of disorders, assess the overall health of an animal, and reflect metal concentrations in the animals environment. Collecting a sufficient number of individuals to determine an analyte reference range for wildlife species can be an especially difficult task. This study reports liver metal concentration ranges for As, V, Cr, Mn, Fe, Ni, Co, Cu, Hg, Se, Zn, Mo, Cd, Ba, Pb and Tl in 32 greater sage-grouse (Centrocercus urophasianus). Sage-grouse sent to the Wyoming State Veterinary Laboratory for diagnostic evaluation were subsequently used in this study. Birds were received from 12 different locations throughout the state, as well as 3 locations in Montana. All birds received a complete diagnostic evaluation, and in addition, liver metal concentrations were determined using Inductively Coupled Plasma Mass Spectrometry. All results for Hg, Tl and As were below their respective detection limits of 0.05, 0.05 and 0.5 mg/kg tissue wet weight. Most values for V and Co were below or slightly above their detection limits 0.1 and 0.05 mg/kg, respectively. Remaining analytes were above the detection limit (0.05mg/kg tissue wet weight) and highly variable.

RP099 (ISA-1117-832329) Avian consumption and use of acid metalliferous water: Toxicological assessments of exposure, effects and susceptibility.
Start time: 8:00 AM
Isanhart, J, MacRae, R², Roberts, C¹, Hooper, M¹, ² US Fish and Wildlife Service, Albuquerque, NM, United States¹ Texas Tech University, Lubbock, TX, United States
Drinking water is a necessity for most birds during annual migration. When migration routes pass through arid environments, water supplies are often in low supply and birds are forced to rely on what few water sources may exist. Industrial and municipal development, agriculture, and other human-induced alterations of the environment, provide avifauna with alternative water sources other than perennial streams, ponds, and springs. Some of these alternative sources provide relatively clean water for consumption and use; however, other water generated from mine waste, oil and gas facilities, and other industrial and commercial processes contain contaminants, sometimes at elevated concentrations, that could result in adverse effects to birds using them. Reports concerning isolated incidents in the western United States have indicated that some of these water sources, specifically acidic, metal rich mine tailings ponds, are lethal to waterfowl, shorebirds, and passerines if ingested. Data indicate that migratory-stressed birds, often in a fasted and/or dehydrated physiological state, may be more sensitive to the acute effects of tailings water. The purpose of our studies was to address the following topics: 1) the influence of physiological status on the acute toxic properties of synthetic acid metalliferous water (AMW) to mallard ducks 2) the potential role that clean water availability and consumption might play in protecting mallard ducks after a toxic exposure to AMW 3) the potential for alleviation of AMW's avian toxic effects by neutralizing its acidic character with lime and 4) the ability of study birds, when given the choice of a number of sources of drinking water, to find a clean water source in the presence of contaminated water. Data are presented on synthetic acid mine water components, behavioral and toxicological observations, mallard water consumption, histopathological findings, and tissue metal concentrations.

RP100 (HUN-1117-832244) Development of extrapolation factors for toxicological effects between exposure media in plant and earthworm studies.
Start time: 8:00 AM
Hunsicker, A¹, Newell, P¹, Podolsky, J¹, ¹ Environmental Health Associates, Inc., Albuquerque, NM, USA
Toxicity reference values (TRVs) are developed for screening-level ecological risk assessments (SLERAs) through a primary toxicity study evaluation process. Adverse effects for plants and invertebrates are based on the concentration of a chemical of potential concern in soil because the primary exposure route is root uptake and oral/dermal contact, respectively. However, the bioavailability and ultimately the toxicity of a chemical to plants and invertebrates is influenced by the soil organic matter (OM) content or type of exposure medium (e.g., filter paper, nutrient solution). Consequently, soil TRV data sets are typically limited to studies using exposure media similar to the soils at the site being studied. As a result, studies and data are eliminated from the literature review if the exposure is in artificial media or in soils with OM content that is not or cannot be discerned as similar to that of the site soil. This exclusion results in limited data sets that lead to an incomplete toxicological assessment of chemicals by a SLERA. The purpose of this poster is to present an evaluation of the relationship between chemical toxicity and exposure media type in terrestrial plant and earthworm toxicity studies. Chemicals with toxicity data in multiple media types were evaluated and used to develop specific factors for the chemical or chemical group (i.e., PCBs and metals) that extrapolate toxicity results from one exposure media type to another. Use of these factors in developing TRVs from previously excluded toxicity data will provide a more complete toxicological assessment for the site of concern. An assessment of the suitability of using these factors in the development of TRVs is also provided to gauge the value of further research to continue this evaluation for other chemicals or chemical groups.

RP101 (RAB-1117-837749) Development of a screening phyto-toxicity test for drilling fluids use for onshore drilling.
Start time: 8:00 AM
Rabke, S¹, ¹ M-I SWACO, Houston, TX, USA
The use of a toxicity test as a regulatory tool for drilling fluids used in offshore drilling activities has been used since 1985 in the Gulf of Mexico. The use of a toxicity test as an environmental performance standard has promoted many advances in drilling fluid technology through the use of innovative chemistry. However, onshore no such standard toxicity test exists for drilling fluids and the current focus regulating onshore discharges using chemical and physical parameters is not promoting advanced drilling fluid technology. The use of a phyto-toxicity test may prove to be a useful tool to evaluate the environmental performance of drilling fluids and begin to move the regulatory framework away from waste disposal toward promotion of beneficial reuse of drill cuttings as soil. Such a test would have to show the appropriate sensitivity to various know chemistries that are used in drilling fluids. One possible test is a seed germination and root elongation test. This test is a short-term screening test that can be easily performed. This study will present data on the evaluation of seed germination and root elongation tests to various common drilling fluid additives.

RP102 (ROY-1117-821196) Toxicity reference values for inhalation of volatile organic compounds by terrestrial mammals.
Start time: 8:00 AM
Lee, B¹, Roy, M¹, ¹ URS Corporation, Austin, Texas, USA
Risk to burrowing mammals from volatile organic compounds (VOCs) in subsurface soil can be significant even though risk to terrestrial mammals above ground may not. The concentrations of VOCs in burrows can be much higher than in the above ground ambient air where atmospheric dilution and dispersion occurs. At a military airfield in the American Midwest, burrowing mammals were a potential receptor to VOC releases. Inhalation toxicity reference values (TRVs) were derived for an ecological risk assessment. Primary toxicity literature was and reviewed for information on ecologically relevant endpoints (reproduction, development, survival, and growth). Relevant toxicity values were used to calculate a daily dose. The daily dose for inhalation is equal to chemical air concentration (ppm) times chemical molecular weight (g/mole) times mammal inhalation rate (m3/day) divided by 24.45 (mol/L) times mammal body weight (kg). Uncertainty factors were applied when necessary to extrapolate a chronic no observed adverse effect level (NOAEL) or a chronic lowest observed adverse effect level (LOAEL) from an acute (LD50) or subchronic LOAEL or chronic LOAEL dose. The chronic NOAEL and LOAEL were used to develop TRVs for the following VOCs: 1,1-dichloroethene, 1,2,4-trimethylbenzene, 2-butanone (methyl ethyl ketone), acetone, carbon tetrachloride, chloroform, ethylbenzene, naphthalene, tetrachloroethene, and total xylenes. Hazard quotients (HQs) based on NOAEL and LOAEL TRVs were calculated each VOC hazard assessment for the thirteen-lined ground squirrel (Spermophilus tridecemlineatus), the ecological receptor. HQ analysis was able to demonstrate there was not unacceptable hazard concern to burrowing mammals for residual VOCs in soil.

RP103 (AAA-1117-468903) Ecotoxicitiy of main gasoline components to Asian earthworm Perionyx excavatus.
Start time: 8:00 AM
An, YJ¹, Jeong, S², ¹ Konkuk University, Seoul, South Korea² Kunsan National University, Department of Environmental Engineering
Soil pollution by gasoline has become a significant environmental problem due to its adverse eco-effects. Toluene and methyl tert-butyl ether (MTBE) are major volatile organic compounds (VOCs) of gasoline. Due to their high volume percentage in fuel as well as their physicochemical properties, they are commonly found in gasoline-contaminated soils. Toluene is known to be toxic to a range of biota. MTBE is classified as a potential human carcinogen by U. S. EPA. They are released into environment not as a single component, and often found as a mixture. It is important to know the influence of main gasoline components such toluene and MTBE to soil organisms, which can be initially exposed to those compounds as a result of the leakage of gasoline from underground storage tanks. There is little information available on the toxicity of toluene and MTBE to soil biota. In this study, the comparative and combined toxicities of toluene and MTBE to the Asian earthworm, Perionyx excavatus were evaluated. Earthworms have been widely used as bioindicators for soil ecotoxicity assessment, and Perionyx excavatus was reported to be a sensitive earthworm species. The median lethal concentration (LC50) was measured to determine the toxic levels of toluene and MTBE individually. The combined effect of toluene and MTBE was investigated using toxic unit model. It was found that toluene was more toxic than MTBE, and the combined toxicity was almost additive to P.excavatus. Treatment with toluene and MTBE also affected the behavior and morphology of P. excavatus. The morphological abnormalities included coiling, curing, mucous secretions, swelling, thinning and fragmentation. It appeared that the VOCs induced metabolic and functional damages in earthworms.

RP103b (SQU-1127-858892) Impacts Of The Effluents Of A Thermal Power Plant On The Chlorophyll Pigments And The Biomass Of Mustard Crops.
Start time: 8:00 AM
Squib, M¹, ¹ DEPARTMENT OF BIOLOY, FACULTY OF NATURAL SCIENCES, TURKEYEN CAMPUS, GEORGETOWN, GUYANA
Oxides of sulphur, nitrogen and carbon particulates are the major air pollutants by the thermal power plant complex of kasimpur (Aligarh, U.P, India) running on 3192MT of coal/day. The effect of these gaseous pollutants was studied on 110 days old Mustard crops (Brassica juncea, variety-T59), growing at 0.5, 2, 4, 6, and 20 Km leeward from power plant complex. These gaseous pollutants induce a significant reduction in the biomass of root and shoot and chlorophyll pigments up to a distance of 4 Km in comparison to the reference site at 20Km leeward from the source of pollution. The significant percent reduction in chlorophyll a (30%), chlorophyll b (27%) and total chlorophyll (28%), causing a serious loss in the root biomass (86%), shoot biomass (83%) and total biomass (83%) of the crops. The loss in chlorophyll contents and biomass showed a significant and positive correlation with the distance from the source of pollution. A significant correlation (r = 0.89) also existed between the total chlorophyll and the total biomass of the crops. The data obtained in the present work indicates that the degree of response in the Brassica juncea increased with the decreasing distance from the source of pollution.

RP103c (COE-1122-405963) PCDDs and PCDFs in Small Mammals Foraging in the Tittabawassee River Floodplain, Michigan.
Start time: 8:00 AM
Coefield, S¹, Zwiernik, M¹, Seston, R¹, Fredricks, T¹, Moore, J¹, Tazelaar, D¹, Kay, D², Jones, P², Bradley, P¹, Giesy, J¹, ¹ National Food Safety and Toxicology Center, Center for Integrative Toxicology, Zoology Department, Michigan State University, East Lansing, MI, USA² ENTRIX, Inc, Okemos, MI, USA
The Tittabawassee River located in central Michigan is known to contain elevated concentrations of polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) in sediments, floodplain soils, and fish downstream of Midland, MI. The Aquatic Toxicology Laboratory at Michigan State University is investigating the impact of this contamination on the terrestrial ecosystem, using the Great Horned Owl (GHO) and insectivorous shrews as indicator species. Mammals, including herbivorous small mammals (chipmunks, mice, voles, and flying squirrels) and insectivorous shrews were sampled at two locations upstream and four locations downstream of Midland from 2003-2004. The results from the mammal data will be used in part to extrapolate GHO dietary exposure to PCDF/DDs. Total concentrations of seventeen 2,3,7,8 PCDD/PCDF congeners, expressed as avian TEQs, ranged from 0.228-4.61 ng/kg, ww, for herbivorous small mammals and 0.38-25.3 ng/kg, ww, for insectivorous shrews at the reference area. Avian TEQs downstream of Midland ranged from 3.48-880 ng/kg, ww, for herbivorous small mammals and 22.9-8790, ww, ng/kg for shrews. The significant difference between the shrew and concentrations in other small mammals is due to their different diets and underlines the importance of dietary composition in determining contaminant exposure.