TP011 (LEE-1122-018410) Risk assessment of heavy metals via exposure pathways of soil, water and crop plant in the abandoned metal mine sites.
Start time: 8:00 AM
Lee, Jin-Soo ¹, Chon, Hyo-Taek¹, ¹ School of Civil, Urban and Geosystem Eng., Seoul National University, Seoul 151-744, Korea, South Korea
Current assessment models derive the total human exposure by evaluating the fate and transport of toxic elements through exposure pathways such as drinking water, food intake, dust inhalation and hand-to-mouth soil ingestion. In order to investigate the contamination levels of heavy metals and assess the risk of adverse health effects on human exposure to heavy metals influenced by past mining activities, environmental geochemical surveys were undertaken around two abandoned metal mine sites (Dokok Au-Ag-Cu and Hwacheon Au-Ag-Pb-Zn mines). Tailing, soil and crop plant samples in the vicinity of the Dokok and the Hwacheon mines were collected from agricultural land around the mine sites. Rice samples were taken from paddy fields. Groundwaters used as a drinking water were collected around the tailings in these mine areas. After appropriate sample preparation, these samples were analyzed for As, Cd, Cu, Pb and Zn by ICP-AES and ICP-MS. High concentrations of heavy metals were found in tailings from the Dokok (98.2 Cd mg/kg, 2,550 Cu mg/kg, 4,200 Pb mg/kg, 18,020 Zn mg/kg) and the Hwacheon (12.4 Cd mg/kg, 580 Pb mg/kg, 1,300 Zn mg/kg) mines. These significant concentrations can impact on soils and waters around the tailing dumps. Elevated levels of As, Cd, Cu, Pb and Zn were also found in agricultural soils from these mine areas. Non-cancer (toxic) risks are indicated in terms of a hazard quotient (H.Q.). H.Q. is ADD (average daily dose)/RfD (reference dose). A toxic risk exists for H.Q.>1. To calculate the hazard index (H.I.) the ADD from three identified pathways (soil, groundwater and food (rice grain) pathways) compared to the relevant RfD obtained from the US-EPA database IRIS is summed. The hazard index of As in the Hwacheon mine area was higher value more than 1.0. Therefore, toxic risk for As exists via exposure (ingestion) of contaminated soil, water and rice grain in this mine area. The cancer risks of being exposed to As by the rice ingestion and drinking water routes in the Hwacheon mine area are 1E-3 and 1E-4, respectively. The As cancer risk via exposure pathways of rice and drinking water exceeded the acceptable risk of 1 in 100,000 for regulatory purposes. Thus, the daily intake of rice and water by the local residents can pose a potential health threat due to long-term arsenic exposure.

TP013 (ARN-1117-824716) Potential effects of commonly used synthetic sea salts on copper toxicity tests using Mytilus.
Start time: 8:00 AM
Arnold, W¹, Cotsifas, Jeffrey², Playle, Richard³, ¹ Copper Development Association Inc., New York, NY, USA² Pacific EcoRisk Inc., Martinez, CA, USA³ Department of Biology, Wilfrid Laurier University, Waterloo, ON, Canada
The salinity of effluents, ambient, and laboratory water samples often requires adjustment to meet guidelines for testing of marine species. Sample salinity is often increased via addition of synthetic sea salts. The US Environmental Protection Agency in its saltwater test guidelines specifically mentions that commercial sea salts, such as Forty Fathoms® (now Crystal Sea Marinemix®, Bioassay Grade), HW Marinemix® or equivalent salts can be used for salinity adjustment. This study focused on the use of synthetic sea salts and their potential effects on toxicity tests of samples where copper is of concern. Thirty-two commercial toxicity testing laboratories in Canada and the USA were surveyed, 27 of which use artificial sea salts. The proportion using Crystal Sea Marinemi® (Bioassay Grade), HW Marinemix®, Instant Ocean® and Tropical Marin® were 56%, 22%, 11%, and 11%, respectively. Saline solutions (30 g/L) were made by adding each sea salt to reverse osmosis deionized water. In addition modified GP2, Kent Marine®, and Red Sea Salt® were similarly prepared. All solutions were analyzed for dissolved organic carbon (DOC) and copper concentrations. Crystal Sea Marinemix® had elevated DOC concentrations (5.4 to 6.4 mg C/L) while other brands generally contained <1 mg/L DOC. Some salt solutions at 30 g/L contained >2.0 g dissolved Cu/L. Dissolved copper EC50s for Mytilus sp. (probably M. galloprovincialis) were determined in saline solutions made from Crystal Sea Marinemix® (9.7 g Cu/L), HW Marinemix® (8.6 g Cu/L) and GP2 (7.5 g Cu/L). Based on a model developed by Arnold (2005), natural DOC at levels found in Crystal Sea Marinemix® should have increased the dissolved copper EC50 to approximately 30 g Cu/L. Excitation-emission spectral analysis and toxicity testing indicate that the DOC in Crystal Sea Marinemix® differs considerably from natural DOC, which could explain the low level of influence of the DOC on copper toxicity.

TP014 (WEB-1117-744866) Effects of Multiple Mining and Municipal Effluents on Fish from Junction Creek, Sudbury, Ontario. 1. Basic Environmental Effects Monitoring (EEM).
Start time: 8:00 AM
Weber, L¹, Dube, M^{1, 2}, Rickwood, C^{1, 2}, Driedger, K¹, Portt, C³, Janz, D¹, ¹ University of Saskatchewan, Saskatoon, SK, Canada² National Water Research Institute, Environment Canada, Saskatoon, SK, Canada³ Cam Portt and Associates, Guelph, ON, Canada
The INCO mining site in Sudbury, Ontario, Canada is highly confounded with three metal mining-related discharges potentially affecting the Junction Creek system in a cumulative manner (Garson, Nolin and Copper Cliff wastewater treatment plants (WWTP)), as well as urban run-off and municipal WWTP discharges. Nonetheless, collecting resident fish is an important first step in monitoring for potential deleterious effects of the effluent discharges under the Environmental Effects Monitoring (EEM) program through the Fisheries Act of Canada. Basic EEM end-points (metal body burdens, age, body weight, length, liver weight, gonad weight, egg size and fecundity) were measured in adult fathead minnow (Pimephales promelus) and creek chub (Semotilus atromaculatus) collected in May 2004. Metal body burdens of Cd, Cu, Rb, La, Se and Sr were elevated in one or both species in the current study in fish collected downstream of metal mining effluent inputs to Junction Creek compared to those from reference sites. On the other hand, body burdens of Al, As, Co, Eu, Ga, Hg, U, V and Zn were decreased in one or both species from metal mining effluent-exposed sites. There were no significant differences among sites in overall fish length or body weight. However, length-at-age was significantly lower in male and female creek chub collected downstream of Garson and Nolin WWTP compared to reference sites. Liver weight was significantly increased in male fathead minnows collected downstream of Garson and Copper Cliff WWTP, while gonad weight was significantly decreased in creek chub collected downstream of Garson WWTP. Comparing all basic EEM end-points in the current study, the ones that were the most sensitive were reproductive (egg size and fecundity). Future studies and monitoring of metal mining effluent should focus on these and other sub-organismal reproductive or early life stage survival end-points.

TP015 (WEB-1117-746038) Effects of Multiple Mining and Municipal Effluents on Fish from Junction Creek, Sudbury, Ontario. 2. Investigation of Cause.
Start time: 8:00 AM
Weber, L¹, Dube, M^{1, 2}, Rickwood, C^{1, 2}, Driedger, K¹, Portt, C³, Janz, D¹, ¹ University of Saskatchewan, Saskatoon, SK, Canada² National Water Research Institute, Environment Canada, Saskatoon, SK, Canada³ Cam Portt and Associates, Guelph, ON, Canada
The INCO mining site in Sudbury, Ontario, Canada is highly confounded with three metal mining-related discharges potentially affecting the Junction Creek system in a cumulative manner (Garson, Nolin and Copper Cliff wastewater treatment plants (WWTP)) as well as urban run-off and municipal sewage discharges. After basic morphometric end-points are obtained for Environmental Effects Monitoring (EEM) through the Fisheries Act of Canada, an Investigation of Cause (Phase II) is required. Sub-organismal reproductive and bioenergetic end-points were compared to the basic EEM end-points (see accompanying poster) in the same adult fathead minnow (Pimephales promelus) and creek chub (Semotilus atromaculatus) collected in May 2004 from Junction Creek. Higher 17-estradiol levels were found in female fathead minnows downstream of Copper Cliff WWTP and municipal sewage WWTP discharges (CC/STP), while male creek chub downstream of Garson and Nolin WWTP discharges had lower 11-ketotestosterone levels. Histological analyses revealed that male fathead minnows, but not creek chub, collected downstream of Garson WWTP and CC/STP had less mature gonads and greater testicular cell degeneration, particularly Sertoli cells, compared to reference fish. Female fathead minnows collected downstream of CC/STP and creek chub collected downstream of Nolin WWTP showed elevated ovarian follicle atresia. Vitellogenin, an indicator of exposure to estrogenic contaminants, was significantly elevated (35-fold higher) in male fathead minnow collected downstream of CC/STP compared to reference sites. Liver glycogen did not differ among fish collected from different sites, while liver triglycerides were significantly lower in fish downstream of CC/STP. Overall, gonadal histology produced the most consistent and sensitive response pattern in fathead minnow and creek chub exposed to metal mining effluents. These sub-organismal reproductive and bioenergetic end-points form the basis for the Investigation of Cause for metal mining effluent in Junction Creek.

TP016 (ARN-1117-828746) Probabilistic methods appropriate for estimating site-specific copper criteria — A San Francisco Bay case study.
Start time: 8:00 AM
Warren-Hicks, W¹, Arnold, W², ¹ EcoStat, Inc., Mebane, NC, USA² Copper Development Association Inc., New York, NY, USA
This poster presents new advances in probabilistic methods for incorporating site-specific or regional-specific chemistry into the development of copper criteria, using San Francisco Bay as a case study. In this paper we present methods for deriving and incorporating spatial and temporal variability of copper and water chemistry when deriving site-specific copper criteria. Two approaches for establishing probabilistic risk-based copper criteria in San Franciso Bay are presented. The first is based on the work of Arnold (Integr Environ Assess Manag, 2005) in which dissolved organic carbon is shown to be an important water quality parameter in determining the bioavailability and thus toxicity of copper in saltwater tests with Mytilus sp. Site-specific log normal distributions of copper concentrations in the Bay are derived and used to establish site-specific risk. The second approach enhances the methods adopted by the California Toxics Rule based on site-specific water effects ratios (WERs). Spatial distributions of WERs are shown to be log normally distributed. Using probabilistic methods incorporating spatial and temporal variability, risk-based site-specific criteria are derived for specific sites in San Francisco Bay and compared to existing copper criteria.

TP017 (MAC-1124-478203) Can the unit world approach provide a common and consistent framework for assessing metals and organics?
Start time: 8:00 AM
Mackay, D¹, Webster, E¹, Harvey, C¹, Doyle, P², ¹ Trent University, Peterborough, Ontario, Canada² Environment Canada, Gatineau, Quebec, Canada
There is an obvious incentive to establish a common "playing field" for assessing the toxicity, fate and exposure potential of all chemicals of commerce, including metals and organics. A number of chemical groups are identified and their environmental properties, similarities and differences are discussed, especially with relation to to their environmental toxicity, fate and exposure of all chemicals can be evaluated and compared, at least for the purposes of relative ranking, using unit world models based on the fugacity or aquivalence equilibrium criteria. Examples are given of such assessments for a variety of chemicals including organics and metals and the results are discussed. It is concluded that a common, consistent assessment framework can be developed.

TP018 (BAS-1125-067990) Modulation of gene expression in the spleen of Peromyscus and Rats exposed to lead.
Start time: 8:00 AM
Basu, A¹, Showe, L², Nebozhyn, M², Spotila, J¹, ¹ Environmental Science Program, Dept. of Bioscience and Biotechnology, Drexel University, Philadelphia² Microarray Facility, The Wistar Institute, Philadelphia
The toxic metal lead (Pb++) is a widespread environmental health hazard as well as a special concern for the US Army, which extensively utilizes lead in ammunition. We have employed cDNA microarrays to analyze the effects of acute Pb++- exposure on gene expression patterns in deer mice (Peromyscus maniculatus<\i>), a widely available wild rodent in North America, and laboratory rats (Rattus rattus<\i>). In deer mice we identified novel putative targets of Pb++-mediated toxicity in spleen in the laboratory, including several genes belonging to the metabolic and transport pathways of acetyl-CoA, an important metabolic component in the mitochondrial section of intermediate metabolism. The imbalance in Acetyl-CoA metabolism may limit production of succinyl-CoA, one of the two reactants necessary for the synthesis of -aminolevulenic acid. Lead toxicity is known to inhibit the synthesis of -aminolevulenic acid, a precursor of heme, which leads to anemia. We also determined gene modulation in the spleens of Peromyscus maniculatus<\i> collected from contaminated sites on an Army ammunition plant. Expression of some genes was enhanced and expression of other genes was depressed. We also measured gene expression in laboratory rats, a model organism which has been the subject of many lead toxicity studies. We compared gene expression data with the blood-Pb level of individual mice and rats. This allowed us to identify a set of putative target genes modulated in Peromyscus maniculatus<\i> and laboratory rats due to lead exposure. These data will allow us to create a gene expression database on an environmental toxicant.

TP019 (HER-1126-557889) Low Level Exposure Of Amphiban Ebryos To Aluminium: Tissue Redidue And Resistence To Challenge Conditions.
Start time: 8:00 AM
Perez-Coll, C^{1, 2}, Servant, R³, Castañaga, L¹, Herkovits, J¹, ¹ Consejo Nacional de Investigaciones Científicas y Técnicas, Argentina² Escuela de Ciencia y Tecnología, Universidad Nacional de San Martín³ Comisión Nacional de Energía Atómica
Low-level exposures to physico-chemical agents are common environmental scenarios. The main purpose of this study was to evaluate the possibility to establish links between tissue residue values in free living vertebrate embryos treated with a wide range of low Al concentrations and their response in case of challenge to lethal concentrations of this metal. Bufo arenarum embryos were treated during 14 days with Al in 3 low but gradually increasing Al concentrations. The final Al concentrations were: 2,6 (A), 26 (B), 256 (C) g Al3+/L (in natural fresh water bodies, dissolved Al in the circumneutral pH was found in a range from 1 to 50 g/L). At 48 and 240 hr post treatment the Al uptake was determined with an atomic absorption spectrophotometer resulting in A: 13 and 14.25; B: 13.45 and 12.5; C: 13.5 and 30 g/mg embryo (ww) respectively. Batches of treated embryos plus their controls were challenged with 3.4, 3.7 and 4 mg Al3+/L. The results reported as lethality up till 7 days post challenge point out a significant beneficial effect (higher survival) in the embryos acclimated with a range of concentrations similar to those found in unpolluted fresh water. The results allow concluding that: i) the Al uptake for exposure scenarios bellow and up to values found in unpolluted freshwater achieves its maximal value within the initial 48 hr of exposure and seems not to be related to the Al concentration in the media (the Bioconcentration Factors range from around 80,000 to 800 depending on treatment); however higher exposure to Al resulted in a significant increase in the Al uptake in the embryo, and, ii) the acclimation to Al resulted in an increased resistance to this metal but only in case of treatments with concentrations around the values found in natural fresh water. Thus, Bufo arenarum embryos seems to be very well adapted to Al as found in unpolluted fresh water bodies while their tissue residue values reflect the Al in the media only for concentrations higher than those found in unpolluted freshwater.

TP020 (HER-1126-558375) Oxidative stress in amphibian embryos exerted by means of physico chemical agents.
Start time: 8:00 AM
Herkovits, J¹, Pérez-Coll, C¹, Fridman, O¹, D´Eramo, J¹, Castañaga, L¹, Stockert, J^{1, 2}, ¹ Instituto de Ciencias Ambientales y Salud, Fundación PROSAMA, Buenos Aires, Argentina² 2Departamento de Biología, Facultad de Ciencias, Universidad Autónoma de Madrid, Cantoblanco, Madrid, España
A large number of physico chemical agents such as UV-B irradiation, metals and pesticides produce reactive oxygen species (ROS: singlet oxygen, superoxide and hydroxyl radicals, H2O2) imposed a strong selective pressure which favoured organisms not only able to prevent the adverse effects of oxidative stress, but also to use O2 as the final receptor of electrons in the respiratory chain. ROS are produced physiologically and they participate in metabolic signalling pathways that regulate cell survival, proliferation and apoptosis. However, they are also generated by a large number of xenobiotics and toxic agents. This "good/bad" balance of ROS is highly relevant for living organisms, and it is often surpassed by external conditions that induce oxidative stress, resulting in damage and death. The adverse effects of oxidative stress on Bufo arenarum embryos at stage 25 (complete operculum) were studied using a wide spectrum of chemical compounds and physical agents. In this work, we describe different experimental conditions that show the possibility (i) to prevent, or at least, to mitigate the adverse effects of oxidative stress, and (ii) to exacerbate the toxicity by means of the synergic effect of agents inducing oxidative stress. In the first case, the photodynamic treatment (generation of ROS in the presence of a photosensitizer, O2 and visible light) exerting lethal effects on the embryos were prevented by previous incubation of the animals with well-known antioxidants (glutathione, ascorbic acid; 1 mM for 48 h). Likewise, the lethality induced by ultraviolet B light (UVB) was prevented with Zn and Se in single or combined pretreatments. Zn ions also protected against the lethality produced by Cu, Al, Cd, Ni, and Pb ions. For the synergic response amphian embryos were subjected to photodynamic treatments plus Zn, UV-B plus Ni and 2,4-D plus Cu resulting in higher lethality than expected from single treatments. The results point out the possibility to evaluate by means of the AMPHITOX test a large number of physico chemical agents with adverse or beneficial effects in relation to oxidative stress as a main mechanisms of toxicity

TP021 (HER-1126-558812) Stage-Dependent Susceptibility To UV-B Radiation In Bufo arenarum Embryos.
Start time: 8:00 AM
Castañaga, L¹, Pérez-Coll, C¹, Herkovits, ¹, ¹ Instituto de Ciencias Ambientales y Salud, Fundacion PROSAMA Paysandu 752
The depletion of the stratospheric ozone layer causes great concern due to the higher intensities of UV-B radiation on living organisms and seems to be one of the factors related to the decline of amphibian populations worldwide. The main purpose of this study was to evaluate the susceptibility of Bufo arenarum embryos at different developmental stages to UV-B radiation. Groups of ten embryos by duplicate at the developmental stages of Blastula (B), Gastrula (G), Neurula (N), Tail bud (TB), Gill circulation (GC), Opercular fold (OF) and Complete operculum (CO) were treated with a range of UV-B radiant exposure from 547.2 to 27360 J/m2 (administered within 100 min.) and mortality was evaluated from 24 hours up to 168 hr post exposure. The LD10, LD50 and LD90 values were obtained and plotted as isolethality curves. Malformations exerted by UV-B were also evaluated. The LD10, 50 and 90 values in J/m2 at 168 hr post exposure were: B: 8552; 18931 and 41902; G: 5022, 10157, and 20539; N:17960; 22176 and 27381; TB: 3539, 5235 and 7744; GC: 1793, 2307 and 2994; OF: 2542, 3180 and 3979; CO: 2003, 2829 and 4036. As an average most of the embryos died within the acute post treatment period. Malformations were obtained in all developmental stages but UV-B resulted teratogenic (Teratogenic Index higher than 1.5) in N, GC and CO. GC stage resulted the most susceptible to UV-B as irradiation ranging from 547.2 to 1641.6 J/m2 exerted severe malformations in 100% of the treated embryos. The most common alterations were: persistent yolk plug, bifid spine, reduced body size, delayed development, asymmetry, microcephaly and anencephaly, flexure in tail and body toward the irradiation side, agenesis or partial gill development, abnormal pigment distribution, hypermotility. The results allow to conclude that after a very high UV-B resistance at early developmental stages (e.g. blastula-gastrula and neurula), the susceptibility of Bufo arenarum embryos to this agent increases very significantly for both lethality and malformations.

TP022 (HER-1126-559098) Evoecotoxicology#: Living Organisms As Biomarkers Of The Coevolution With Environmental Agents During The Evolutionary Process.
Start time: 8:00 AM
Herkovits, J¹, ¹ Instituto de Ciencias Ambientales y Salud, Fundación PROSAMA, Paysandu 752
Life forms interacted with changing environmental conditions during almost 4,000 million years. For most of the evolutionary process our understanding of both the environment and life forms is extremely limited and controversial. This article focuses on the hypothesis that living organisms (mainly) at developmental stages could be considered as biomarkers of the coevolution of life forms and environmental features during the evolutionary process. In this context the (almost) anaerobic metabolism at blastula stage embryo could reflect the existence of multicellular organisms in the deep anoxic Earth while the transition to aerobic metabolism at gastrula could be related to the rise in free O2 around 2 billion years ago and the evolution towards tridermic organisms. Complementarily the high resistance to physico chemical agents at early developmental stages could reflect that living organisms had to cope with very adverse environmental conditions during the initial 2 billion years (including UV-B irradiation) while the high susceptibility at developmental stages with complex cell differentiation and morphogenetic processes (e.g. organogenesis and metamorphosis) contribute to the vision that those cell activities were achieved in times of low environmental stress conditions. Taking into account that also mass extinction events could be associated to chemical bomb scenarios (Herkovits 2001), the resistance at developmental stages to physico chemical agents could contribute to the understanding of the environmental stress supported by any specie during its phylogenesis. Evoecotoxicology by considering living organisms as biomarkers of the interactions with chemical and physical agents during the evolutionary process could contribute to a better understanding of the evolutionary process in the Earth. # Evoecotoxicology: The science concerned with the register in living organisms of the interactions with chemical and physical agents during the evolutionary process.