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Alabama Academy of Science Symposium, 2006 "Southeastern Society for Environmental Toxicology and Chemistry" Annual Meeting, Troy University.

THE EVALUATION OF CRYO-PROTECTION ON EMBRYOS OF XENUPUS LARVA. Marie Pierre-Louis, J.S.U., Jacksonville, AL 36265. Dr. Cline, Dept of Biology, J.S.U., Jacksonville, AL 36265.

The use of cryo-preserving embryos of Amphibians could be useful in toxic testing. This research focused on the evaluation of cryo-protection on Xenopus embryos. Five solutions (2 different kinds of ethylene glycol, Galactose, DMSO, and PVP) were used as cryo-protection. Twenty embryos were used for each treatment. For each treatment, embryos were placed in a Petri dish containing 8ml of the cryo-protection solution plus one control solution for 5 minutes. . Twenty embryos were placed into 8M of ethylene glycol than replaced into 2M of ethylene glycol. The same amount of embryos was placed into galactose, DMSO and PVP. Then the embryos were placed into plastic straws capped with parafin. Those straws were then placed in a -20[degrees]C freezer until thaw. After 3 days, the embryos were unfrozen by swirling them gently into a water bath for about one minute. They were then placed into a Petri dish containing control solution, and observed. Embryos in the DMSO, galactose, and ethylene glycol lysised. For the PVP, nineteen of those embryos were recognizable but had some deformation. Thirty minutes later, the eggs started to reshape. Some cells were alive but not the embryos. The PVP was seen to be the best cryo-protection but more tests including toxicity testing need to be done. This experiment was supported by the Alabama Academy of Science research award.

ANALYSIS OF ALKYLPHENOLS IN WASTEWATER USING SOLID-PHASE EXTRACTION AND GAS CHROMATOGRAPHY/MASS SPECTROMETRY. Yassin A. Jeilani, Bianca M. Coley and Victor M. Ibeanusi, Spelman College, Environmental Science and Studies, Atlanta, GA 30314.

Interest in the health effects associated with alkylphenols has made the analysis in environmental matrices a priority. Solid-Phase Extraction (SPE) method was developed for the extraction of selected alkylphenols in wastewater followed by derivatization and analysis by gas chromatography/mass spectrometry in single ion monitoring mode. The analytical method was used in the analysis of influent and effluent samples collected from a local wastewater treatment plant. The effect of sample pH, SPE sorbent and sample volume used in the extraction were compared in terms of recovery data.

PHYTOTOXICITY OF MONOMERIC AND POLYMERIC ALUMINUM TO NATIVE WETLAND PLANT SPECIES. Ivan Stefanov and Marina Koether, Department of Chemistry and Biochemistry, Kennesaw State University, Kennesaw, GA 30144. Heather Sutton, Department of Biological and Physical Sciences, Kennesaw State University, Kennesaw, GA 30144.

The most commonly used coagulant for the treatment of drinking water is aluminum sulfate (alum). The aluminum (Al) species in alum is the monomeric species A[l.sup.3+] (Al([H.sub.2]O[).sub.6.sup.3+]). However, polymeric Al coagulants such as polyaluminum chloride used to treat drinking water may contain up to 10% of the Al as the polymeric species, A[l.sub.13] [.sup.7+] ([Al[O.sub.4]A[l.sub.12](OH[).sub.24]([H.sub.2]O[).sub.12].sup.7+]). These Al species can enter the environment through the land farming of the aluminum hydroxide sludge produced during the sedimentation process in the treatment of drinking water. Aluminum in this sludge can then enter waterways and wetlands through leaching or runoff. The toxicity of Al [.sub.13.sup.7+] in the environment relative to Al [.sup.3+] is in question. The goal of this study was to investigate the relative toxicity to wetland plants of Al [.sub.13.sup.7+] as compared to Al [.sup.3]. Native wetland plant species tested were swamp milkweed (Asclepias incarnata) and saw-tooth sunflower (Helianthus grosseserratus). Additionally, radish (Raphanus sativus) was tested, as it is an agricultural species commonly used as a surrogate in place of native plants. Concentrations ranging from 0 to 500mg/L were evaluated. Endpoints measured were shoot and root length and mass, as well as internal plant concentrations of aluminum and various nutrients. Despite high variability in the data, a number of patterns emerged. Radish showed more differences in response between the two aluminum species than the native plant species did. With some of the internal plant aluminum or nutrient concentrations, the two aluminum species caused a different pattern of response.


Perchlorate is a highly water soluble anion that moves through ground and surface waters, accumulating rapidly in the environment. It is used as an oxidizer in solid propellants, ballistics, in rocket and missile fuels, fireworks, munitions. Lakes and underground waters near rocket-assembly plants and military bases were shown to have high level of perchlorate. When released into the environment, perchlorate becomes an environmental pollutant posing immunotoxic conditions in animals and possibly in plants. The toxic effects of most pollutants are believed to be exerted on key metabolic enzymes, on photosynthetic apparatus, on growth, yield and quality. The objectives of this research are to determine the effects of perchlorate contamination on the plants' photosynthetic abilities and to investigate the involvement of perchlorate in oxidative stress induction in Arabidopsis thaliana, sorghum bicolor, and Nicotiana tobaccum. Plants were grown and maintained in potted soil in a growth chamber and then screened for toxicological responses when challenged with perchlorate solutions. Chloroplasts were isolated by differential centrifugation and assayed for the light reactions of photosynthesis. Isolated cytoplasmic proteins and chloroplasts were assayed for SOD activities. Plant growth, biomass accumulation were also recorded. The data showed that sorghum and tobacco responded to perchlorate by shifting to underground growth. Higher SOD activities and reduced NADP[H.sub.2] were also recorded in perchlorate treatments.


Waters and soils in Anniston, AL are contaminated by PCB-laden runoff from landfills at the local Monsanto plant. Despite clean-up efforts, soils, waters and the blood of local residents still have elevated levels of PCBs. In order to assess the effects of current PCB contamination on aquatic organisms, we are analyzing the development of the frog Xenopus laevis when exposed to control solutions, waters and aqueous sediment extracts from streams near the Monsanto site using both the standard Frog Embryo Teratogenesis Assay--Xenopus (FETAX) and extended incubations of frog embryos in control solutions and test samples. The standard FETAX assay shows subtle but significant growth inhibition in embryos exposed to contaminated waters and sediment extracts, though mortality and malformation rates are not affected. Because anuran embryos are relatively insensitive to the effects of PCBs and dioxins during the first five days of development and become more sensitive later, we are developing an extended modification of the FETAX assay where embryos are exposed to samples for two weeks or more. Preliminary results indicate embryos raised in contaminated Snow Creek water for 24 days show significant growth inhibition compared to the controls. We are continuing extended incubation assays over the first 2-4 weeks of development and are plan to develop a similar assay using zebrafish as a new model organism.

GERONTOGENE EFFECTS ON THE RESISTANCE OF NEMATODES TO HYDROGEN PEROXIDE OR NICKEL TOXICITY. Jeffrey Zuiderveen, Joanne Brown, Dept. of Biology, Columbus State Univ., Columbus, GA 31804 and Aisha Kelly, Ga. Perimeter College, Conyers, GA 30012.

Gerontogenes are genes that, when mutated, cause an increase in the lifespan of the organisms beyond that of the wild-type. Several of these genes have been identified in the nematode Caenorhabditis elegans. Among these are the clk-1 and the daf-2 mutations. Although they affect separate pathways, nematodes with either one of these mutations will live twice as long as the wild-type, and those with both mutations have double the longevity of the single mutations. To investigate the gerontogene effects on toxicity, nematodes were subjected to varying concentrations of toxicants (hydrogen peroxide, nickel chloride) that utilized different mechanisms of toxicity. Survival was observed for 1-2 weeks, depending on the toxicant. Results seemed to indicate that these mutations tended to make the nematodes slightly less susceptible to hydrogen peroxide, but quite a bit more sensitive to nickel. Therefore, while these gene mutations may possibly seem desirable to researchers who seek a "Fountain of Youth" for humans, the nickel tests should give pause if those striving for greater longevity don't wish to live in a closed environment.

COMPARATIVE ACUTE TOXICITY OF HEAVY METALS TO DRAGONFLY LARVAE. Valerie Tollett and Timothy Rice, Dept. of Biology, Univ. of S. Ala., Mobile, AL 36688.

Dragonflies (Insecta: Odonata) occupy an important position in aquatic ecosystems, serving as both predator and prey throughout their life cycle. These natural history features create the potential for long term exposure to toxicants from a variety of sources. Despite their importance in aquatic ecosystems, there is little field or laboratory data regarding the impact that environmental chemicals have no odonates. To determine the impact of heavy metals on dragonfly larvae, acute toxicity tests were conducted in the laboratory. Larvae measuring 5-10mm were collected from the field and held in a circulating, filtering system for one week. The species, Pachydiplax longipennis, was chosen for this study due to their abundance and wide distribution. Seven day acute toxicity tests were conducted to compare the effects of equimolar concentrations (0, 0.04, 0.35, 0.89, 2.2mM) of lead, copper and cadmium. Using immobility as the endpoint, the larvae exhibited a high tolerance to cadmium and lead, but high levels of mortality to copper. This species of odonate appears to be extremely resistant to metals compared in general to other common aquatic test species (e.g. Daphnia, Chironomus, Rana, Xenopus). High resistance to metal pollution could make odonate larvae a potential indicator of poor water quality in contaminated habitats. Their ability to withstand and potentially accumulate high levels of metals might also put their predators at risk from ingestion of contaminated larvae or adults.

LEAD ASSESSMENT WITHIN THE SOIL, WATER, AND VEGETATION IN SHOOTING RANGES AT THE GRAND BAY NERR. Katherine E. VanDeven and Timothy Rice, Dept. of Biology, University of South Alabama, Mobile, AL 36688.

The Grand Bay National Estuary Research Reserve was established in 1999 by NOAA and is maintained by the Mississippi Department of Marine Resources. The reserve is located in Jackson County, Mississippi between Pascagoula and the Alabama state line. The abandoned Live Oak Rifle and Pistol Range lie within the boundaries of the NERR and is the main study site. The Live Oak Rifle and Pistol Range ecosystem consists of forested areas, wetlands that are seasonal, and a permanent pond. This project addresses the amount of lead present in the Live Oak Rifle and Pistol Range. Four collection sites were chosen throughout the range, and included: a short-range target, a long-range target, a pond behind the range, and the shooting area. The ditch water along the road was also sampled to determine the difference between contaminated road water and the range water. The lead levels for the soil samples ranged from 19.57 [micro]g/g to 137,835.9 [micro]g/g. Lead levels for the vegetation samples ranged from 2.66 [micro]g/g to 2290.37 [micro]g/g. Lead levels in water samples ranged from 0.45 [micro]g/L to 557.4 [micro]g/L; some of these samples were above EPA acceptable limits. These values were comparable to levels reported from other shooting ranges. These initial results indicate possible lead contamination within this estuary reserve.

ACUTE TOXICITY OF FIPRONIL AND ITS ENANTIOMERS TO MARINE AND FRESHWATER NON-TARGETS. Jay Overmyer, Univ. of GA, Dept. of Ent., Athens, GA 30602. A.W. Garrison, USEPA, NERL, Athens, GA 30605. J. Avants, USEPA, NERL, Athens, GA 30605. M. DeLorenzo, NOAA/NOS, Charleston, SC 29412. P. Key, NOAA/NOS, Charleston, SC 29412. K. Chung, NOAA/NOS, Charleston, SC 29412. B. Konwick, Univ. of GA, Dept. of Env. Health Sci., Athens, GA 30602. W. A. Wilson, Univ. of GA, Dept. of Env. Health Sci., Athens, GA 30602. M. Black, Univ. of GA, Dept. of Env. Health Sci., Athens, GA 30602

Fipronil is a phenylpyrazole insecticide used in agriculture and domestic settings for controlling various insect pests in crops, lawns and residential structures. Fipronil is chiral; however, it is released into the environment as a racemic mixture of two enantiomers. In this study, the acute toxicity of the (+) and (-) enantiomers and the racemic mixture of fipronil were assessed using Ceriodaphnia dubia (water flea), Simulium vittatum IS-7 (black fly), Xenopus laevis (African clawed frog), Palaemonetes pugio (grass shrimp), Mercenaria mercenaria (hardshell clam), and Dunaliella tertiolecta (phytoplankton). Results showed that S. vittatum IS-7 was the most sensitive freshwater species while P. pugio was the most sensitive marine species. Ceriodaphnia dubia and S. vittatum IS-7 were the only species that showed greater sensitivity to a specific isomer or the racemate of fipronil. Increased mortality and minimal recovery was observed in all species tested for recovery from fipronil exposure.

DISTRIBUTION OF CRASPEDACUSTA SOWERBII IN CENTRAL ALABAMA OVER THE LAST 3 YEARS. Kaci Rodgers and James Rayburn, Biology Dept., Jacksonville State University 700 Pelham Road North, Jacksonville, AL 36265-1602.

Craspedacusta sowerbii, which is better known as the freshwater jellyfish was first described in 1880. Researchers suggest the freshwater jellyfish may have been transported from pond to pond on the feet of waterbirds. These jellyfish have been found in lakes, farm ponds, rivers, and water-filled quarries. The freshwater jellyfish is an invasive species. It has been found in many different areas in the Southern United States. In Alabama, it has been found in a few areas around Jacksonville State University. The freshwater jellyfish has been reported to live in water with a ph ranging 6.5-8.5. This research is aimed to predict when the freshwater jellyfish might appear by analyzing data logs of when they have appeared. Different areas across Alabama, where these jellyfish have been spotted, will be evaluated to help pinpoint the areas and time of year this species shows up. The data analysis will determine the prime months when jellyfish collection might be significant.

THE DEVELOPMENTAL TOXICITY OF RESMETHRIN TO EMBRYOS OF XENOPUS LAEVIS. James Rayburn, Biology Department, Jacksonville State University, Jacksonville, AL 36265.

Natural pyrethrin insecticides are commonly used as mosquito-cides and are currently being used locally in Jacksonville AL. These insecticides are used during the spring when many amphibian species are breeding and larvae are developing. Because this could be sensitive time for frog species, frogs could be affected by these types of chemicals. Resmethrin was chosen as a representative pyrethrin. Pyrethroid insecticides are known to be neurotoxic to mammals. The developmental effects of pyrethrin on aquatic species (in particular amphibian species) is unclear. The Frog Embryo Teratogenesis Assay Xenopus (FETAX) was used to determine the toxicity of pyrethrin in accordance with the ASTM (1991) procedures. This procedure exposed early blastula embryos to a free living tadpole to the pyrethrin. Mortality, Malformation and embryo length were collected. Data was analyzed using Toxtools. Both pure Resmethrin and a water soluble fraction of a commercial resmethrin were tested. Results indicated that resmethrin does not cause significant mortalities at concentrations >50 ppm. Malformations were observed. Forty-eight hour old Xenopus embryos exhibited a higher startle response than controls, even at concentrations as low as 1 ppm. This study suggested behavioral studies on responses on embryos exposed to resmethrin should be performed.

EFFECTS OF NANOPARTICLES ON GILL FUNCTION. Roxana Weil, Kelly Hyndman,Nancy Denslow and David Barber, UF

Nanomaterials have very high surface areas and high percentages of their component atoms on the surface, which leads them to have unusual characteristics. Because of these characteristics, nanomaterials have many uses including sporting goods, cosmetics, paints, electronics, composites, and propellants. Due to their widespread applications, nanomaterials will inevitably be released into the aquatic environment, where they will interact with aquatic organisms. The unique physical and chemical properties of nanomaterials may also lead to unique biological properties. Unfortunately, toxicity testing of nanomaterials has been extremely limited, and factors that influence the biological properties of nanomaterials are poorly understood. The goal of this study was to determine if aluminum nanoparticles adversely affect fish and if the effects are different than soluble aluminum. Zebrafish (Danio rerio) were exposed to 0, 0.5, 2.5 and 12.5 mg aluminum/L as aluminum chloride or 51 nm spherical aluminum nanoparticles for up to 96 hours. Particles were characterized before and after exposure to determine changes in aggregation and dissolution. No mortality was induced by aluminum nanoparticle exposure, however, exposure to aluminum nanoparticles produced dose dependent reduction of gill Na/K-ATPase activity, indicating that gill function was compromised. To determine if observed effects were due to direct actions on the gill, primary cultures of gill filaments were exposed to soluble or nano-sized aluminum (0-12.5mg/L). Finally, to determine if uptake of intact particles occurs at the gill surface, zebrafish and gill filaments were exposed to insoluble CdSe/ZnS quantum dots for up to 48 hours and examined by fluorescence microscopy. This research was supported by a grant from the National Science Foundation (BES-0540920).

REPRODUCTIVE SUCCESS OF PIMEPHALES PROMELAS EXPOSED TO p,p'-DDE. David S. Barber, Center for Environmental and Human Toxicology and Elizabeth J. Ray, School of Natural Resources and Environment, University of Florida, Gainesville, FL 32608.

p, p'-DDE is the most persistent metabolite of the widely-used pesticide, DDT. Laboratory studies have shown DDE and other endocrine disrupting compounds (EDCs) alter hormone levels and gene expression patterns in the fathead minnow (Pimephales promelas), but have not linked those findings to population-level effects. The goal of this study was to examine the relationship between DDE body burden, expression of standard biomarkers, and reproductive output. The number of eggs laid, percent of eggs fertilized, and percent of eggs hatched as well as survival of offspring were measured in fathead minnows exposed to DDE. A control group (vehicle only) and three treatment groups (1.63 ppm, 11.48 ppm, and 104.25 ppm DDE in fish feed), each containing eight females and four males, was measured for 28 days. Survival rates and growth of offspring laid between 21-28d of the experiment were monitored until the onset of first reproduction. The number of eggs laid in the highest treatment group was reduced by 74% as compared to the control group. Average DDE body burden of fish in the highest treatment group was 110.57 ppm. This experiment suggests DDE exposure does not affect fertilization, hatchability, or fry survival rates, but can reduce the total number of eggs produced by females as well as growth and time to first reproduction of fish exposed in ovo.

THE CONCENTRATION OF THREE METALS IN TISSUES OF SELECT MARINE SPECIES FROM MOBILE BAY Bernice Moser and James R. Rayburn. Dept. of Biology, Jacksonville State University, Jacksonville Al 36265.

Mobile Bay, and estuary in the Gulf of Mexico, has more than 400-point source dischargers of pollutants and eight ancillary water bodies that have been labeled as impaired due to the high levels of toxic chemicals such as pesticides, priority organics, and metals by the Alabama Department of Environmental Management (Peachey, 2003). The ecosystem is important to the fishing industry in Mobile and many of these species are susceptible to metal contamination either by their association with the sediments or by bioaccumulation. To determine an estimate of metal contamination in Mobile Bay selected fish species were bottom collected using the research vessel Verril from Dauphin Island Sea Lab between channel markers 17 and 19. Sample collection began in February of 2005, and fish collected included: gulf menhaden (Brevortia patronus), hardhead catfish (Arius felis), hogchoker (Trinectes maculates), spot (Leiostomus xanthurus), and the least puffer (Sphoeroides parvus). The fish were dissected, liver and muscle tissue removed for metal analysis and frozen until processed. Samples were weighed and using trace-metal grade nitric and hydrochloric acid, digested for graphite furnace atomic absorption spectrometry. The metals examined included lead, copper, and zinc. The data was analyzed using ANOVA to compare between species.
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Publication:Journal of the Alabama Academy of Science
Article Type:Conference notes
Geographic Code:1U6AL
Date:Apr 1, 2006
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