D. George Dixon

Phytoremediation of arsenate contaminated soil by transgenic canola and the plant growth-promoting bacterium Enterobacter cloacae CAL2

It was previously observed that transgenic tomato plants that express the Enterobacter cloacae UW4 1-aminocyclopropane-1-carboxylate (ACC) deaminase (EC 4.1.99.4) gene, and thereby produce lower levels of ethylene, were partially protected from the deleterious effects of six different metals. However, since tomato plants are unlikely to be utilized in the phytoremediation of contaminated terrestrial sites, transgenic canola (Brassica napus) plants that constitutively express the same gene were generated and tested for their ability to proliferate in the presence of high levels of arsenate in the soil and to accumulate it in plant tissues. The ability of the plant growth-promoting bacterium E. cloacae CAL2 to facilitate the growth of both non-transformed and ACC deaminase-expressing canola plants was also tested. In the presence of arsenate, in both the presence and absence of the added plant growth-promoting bacterium, transgenic canola plants grew to a significantly greater extent than non-transformed canola plants.

Toxicity of sixty‐three metals and metalloids to Hyalella azteca at two levels of water hardness

The toxicity of all atomically stable metals in the periodic table, excluding Na, Mg, K, and Ca, was measured in one-week exposures using the freshwater amphipod Hyalella azteca in both Lake Ontario, Canada, and soft water (10% Lake Ontario). Metals were added as atomic absorption standards (63 metals), and also as anion salts for 10 metals. Lethal concentrations resulting in 50% mortality (LC50s) were obtained for 48 of the metals tested; the rest were not toxic at 1,000 microg/L. The most toxic metals on a molar basis were Cd, Ag, Pb, Hg, Cr (anion), and Tl, with nominal LC50s ranging from 5 to 58 nmol/L (1 to 58 nmol/L measured). These metals were followed by U, Co, Os, Se (anion), Pt, Lu, Cu, Ce, Zn, Pr, Ni, and Yb with nominal LC50s ranging from 225 to 1,500 nmol/L (88-1,300 nmol/L measured). Most metals were similarly or slightly more toxic in soft water, but Al, Cr, Ge, Pb, and U were >17-fold more toxic in soft water; Pd was less toxic in soft water. Atomic absorption (AA) standards of As and Se in acid had similar toxicity as anions, Sb was more toxic as the AA standard, and Cr and Mn were more toxic as anions. One-week LC50s for H. azteca correlate strongly with three-week LC50s and three-week effect concentrations resulting in 50% reduction in reproduction (EC50s) in Daphnia magna.

Fathead minnow (Pimephales promelas) reproduction is impaired in aged oil sands process-affected waters.

Large volumes of fluid tailings are generated during the extraction of bitumen from oil sands. As part of their reclamation plan, oil sands operators in Alberta propose to transfer these fluid tailings to end pit lakes and, over time, these are expected to develop lake habitats with productive capabilities comparable to natural lakes in the region. This study evaluates the potential impact of various oil sands process-affected waters (OSPW) on the reproduction of adult fathead minnow (Pimephales promelas) under laboratory conditions. Two separate assays with aged OPSW (>15 years) from the experimental ponds at Syncrude Canada Ltd. showed that water containing high concentrations of naphthenic acids (NAs; >25 mg/l) and elevated conductivity (>2000 μS/cm) completely inhibited spawning of fathead minnows and reduced male secondary sexual characteristics. Measurement of plasma sex steroid levels showed that male fathead minnows had lower concentrations of testosterone and 11-ketotestosterone whereas females had lower concentrations of 17β-estradiol. In a third assay, fathead minnows were first acclimated to the higher salinity conditions typical of OSPW for several weeks and then exposed to aged OSPW from Suncor Energy Inc. (NAs ∼40 mg/l and conductivity ∼2000 μS/cm). Spawning was significantly reduced in fathead minnows held in this effluent and male fathead minnows had lower concentrations of testosterone and 11-ketotestosterone. Collectively, these studies demonstrate that aged OSPW has the potential to negatively affect the reproductive physiology of fathead minnows and suggest that aquatic habitats with high NAs concentrations (>25 mg/l) and conductivities (>2000 μS/cm) would not be conducive for successful fish reproduction.

Pathway of anthracene modification under simulated solar radiation

Exposure of polycyclic aromatic hydrocarbons (PAHs) to sunlight results in rapid structural photomodification generally via oxidation reactions. These PAH modification products are in many cases more toxic than their parent compounds. In this study, anthracene (ANT), a rapidly photooxidized PAH, was irradiated with simulated solar radiation (SSR, 100 micromol m(-2) s(-1)) in aqueous solution to examine the photomodification pathway. The photoproducts formed were identified by HPLC. The ANT product profile after 9 h in SSR was very complex, with more than 20 compounds detected. The photoproducts formed were anthraquinones, benzoic acids, benzaldehydes and phenols showing the process to be oxidative in nature. Some of the anthraquinones were themselves subject to photooxidation, and were thus intermediates in the product pathway. The kinetics of ANT photooxidation revealed a pseudo first-order reaction with a half-life of 2 h under the SSR source used. The kinetics of product formation allowed deduction of a probable photomodification pathway. This study indicates that PAH photooxidation products are likely to exist as complex, dynamically changing mixtures in PAH contaminated aquatic environments.

A holistic approach to ecosystem health assessment using fish population characteristics

The status of a fish population is a reflection of the overall condition of the aquatic environment in which that population resides. As such, fish population characteristics can be used as indicators of environmental health. Simple and inexpensive methods to follow fish population responses to environmental degradation are lacking. This paper outlines a protocol whereby environmental impacts on fish populations are classified by five patterns based on characteristics such as mean age, fecundity and condition factor. The patterns summarize population changes and describe responses to exploitation, recruitment failure, the presence of multiple stressors, food limitation and niche shifts. Classification is best based on the selection, and appropriate sampling, of a comparable reference population. Population characteristics can be used to examine ecosystems exposed to stressors for evidence of long-term damage, and when used with biochemical indicators, can be a powerful tool for ecosystem health assessment. The five responses are illustrated using published data on a number of species challenged by increased predation pressure, acidification, eutrophication, mine waste and reservoir impoundment. Application of this scheme will aid in directing and focusing research efforts on crucial aspects impacted by changing conditions.

Photoinduced toxicity of polycyclic aromatic hydrocarbons to Daphnia magna: ultraviolet-mediated effects and the toxicity of polycyclic aromatic hydrocarbon photoproducts.

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous environmental contaminants known for their photoinduced toxicity. This toxicity may occur through two mechanisms: Photosensitization, and photomodification. Photosensitization generally leads to the production of singlet oxygen, a reactive oxygen species that is highly damaging to biological molecules. Photomodification of PAHs, usually via oxidation, results in the formation of new compounds and can occur under environmentally relevant levels of actinic radiation. The toxicities of 16 intact PAHs to Daphnia magna were assessed under two ultraviolet radiation conditions. The toxicity of intact PAHs generally increased in the presence of full-spectrum simulated solar radiation relative to that in the presence of visible light plus ultraviolet A only. Despite the knowledge of a bipartite mechanism of phototoxicity that includes photosensitization and photomodification, few studies have examined the effects of PAH photoproducts on animals. To expand the existing data, 14 PAH photoproducts (oxy-PAHs) also were assayed, most of which were highly toxic without further photomodification. Two photoproducts of benzo[a]pyrene, 1,6- and 3,6-benzo[a]pyrenequinone, were the most toxic compounds tested, followed closely by benz[a]anthraquinone. Each of these three compounds had a median effective concentration in the low nanomolar range. The data presented highlight the effects of ultraviolet radiation on mediating PAH toxicity and the need to analyze absorption spectra of contaminants in the prediction of photoinduced toxicity. The importance of the role of photomodification also is stressed, because several oxy-PAHs, an unregulated group of contaminants, were highly toxic to D. magna, a key bioindicator species in aquatic ecosystems.

Distribution and Food-web Transfer of Mercury in Napoleon and Winam Gulfs, Lake Victoria, East Africa

Abstract Mercury (Hg) concentrations were measured for the food webs and water of Napoleon Gulf (Uganda) and Winam Gulf (Kenya) in northern Lake Victoria. Water total mercury (THg) concentrations in Lake Victoria range from 1.7 to 5.8 ng/L, while methylmercury (MeHg) concentrations range from 0.2 to 1 ng/L. Water Hg concentrations in Lake Victoria are higher than in temperate great lakes, including Lakes Baikal, Michigan, and Ontario, but the top predator Nile perch have relatively low THg concentrations compared to temperate piscivorous fish. While the water Hg concentrations are similar between Napoleon and Winam gulfs, the THg concentrations in biota are significantly higher in Napoleon Gulf than in the same species from Winam Gulf, which may be due to biogeochemical differences in each gulf. THg concentrations in Nile perch and Nile tilapia consistently increase with total length in both gulfs and the rates of increase are similar. The rates of THg bioaccumulation, as indicated by the regression slopes of log-THg vs. stable nitrogen isotope values for each food web (slopes of 0.163 and 0.165 for Napoleon and Winam gulfs, respectively), are within the ranges of bioaccumulation rates observed in temperate and tropical lakes elsewhere which suggests that Hg bioaccumulates at a similar rate in diverse aquatic food webs, regardless of latitude or species composition.

Acute toxicity, uptake, depuration and tissue distribution of tri-n-butyltin in rainbow trout, Salmo gairdneri

Abstract The tri- n -butyltin cation (Bu 3 Sn + ), the active ingredient in organotin-containing antifouling paints, was highly toxic to rainbow trout ( Salmo gairdneri , mean weight, 1.5 g); the 96-h LC 50 was 1.41 μg Sn/l. Lake trout ( Salvelinus namaycush , mean weight, 5.9 g) were more tolerant, with a 96-h LC 50 of 5.21 μg Sn/l. Rainbow trout concentrated significant levels of Bu 3 Sn + during a 64-d exposure at 0.21 μg Sn/l, with bioconcentration factors of 406 and 570 (based on Bu 3 Sn + and total Sn, respectively). Rainbow trout depurated Sn slowly on transfer to Bu 3 Sn + -free water. During the 32 d period following transfer, the whole-body concentration of Bu 3 Sn + fell by 25% while the total Sn concentration was reduced by 17%. The total Sn concentrations in tissues of rainbow trout at the end of a 15-d exposure to 0.42 μg Sn/l indicated that Bu 3 Sn + partitions into trout on the basis of a three-compartment model. Peritoneal fat (mean concentration, 9.18 mg Sn/kg) constituted one compartment, kidney, liver and gall bladder/bile (mean concentration range, 3.07 to 3.72 mg Sn/kg) a second, and all other tissues (mean concentration range, 0.49 to 1.53 mg Sn/kg) a third. After 15 d of depuration the system had simplified; only two compartments (liver plus gall bladder/bile and all other tissues) were apparent. Varying proportions of Bu 3 Sn + and its metabolites di- n -butyltin, n -butyltin and inorganic tin were present in all tissues sampled. The percent of metabolites in liver (74) and gall bladder/bile (89) were, however, significantly higher than the levels in all other tissues, which ranged from 10 to 43. This suggests hepatic dealkylation and biliary-fecal excretion.

Applications of the aquatic higher plant Lemna gibba for ecotoxicological assessment

Although higher plants represent a significant portion of the total biomass in some aquatic environments, their use in ecosystem evaluation has lagged behind that of other organisms. This is partly due to a lack of convenient aquatic higher plant systems that can be employed for ecotoxicological assessment. However, the aquatic C-3 monocot Lemna gibba has many attributes that makes it useful for ecosystem health assessment. In this report, using examples from the literature and our research, some of the applications Lemna has for environmental research are discussed. Toxicant impacts on Lemna can be readily assessed in terms of growth; the plants multiply quickly and changes in biomass (which doubles approximately every 2 days) can be accurately measured by counting leaves. The plants are small, allowing for simultaneous multiple replication. The small size also makes the lighting conditions easy to control; sunlight can be accurately simulated and specific spectral regions can be enhanced or deleted. Lemna is amenable to in vitro chlorophyll and photosynthesis assays, which make excellent companion endpoints for growth. The plants assimilate chemicals directly from the growth medium, facilitating controlled toxicant application. Furthermore, Lemna has a high bioconcentration capacity, indicating a potential for use in bioremediation technologies.

Sampling-rate calibration for rapid and nonlethal monitoring of organic contaminants in fish muscle by solid-phase microextraction.

Solid-phase microextraction (SPME) is a promising technique for determining organic contaminants within biotic systems. Existing in vivo SPME-kinetic calibration (SPME-KC) approaches are unwieldy due to the necessity of predetermining a distribution coefficient for the analyte of interest in the tissue and the preloading of a calibrating compound to the fiber. In this study, a rapid and convenient SPME alternative calibration method for in vivo analysis, termed SPME-sampling rate (SPME-SR) calibration, was developed and validated under both laboratory and field conditions to eliminate such presampling requirements. Briefly, the SPME probe is inserted into tissue, in this study fish dorsal-epaxial muscle, for 20 min allowing the concentrations of target analytes in the fish muscle to be determined by the extracted amount of analyte and the predetermined sampling rates. Atrazine, carbamazepine, and fluoxetine were detected nonlethally in the low ppb levels within fish muscle, with both laboratory and field-derived results obtained by in vivo SPME-KC comparable (within a factor of 1.27) to those obtained by lethal sampling followed by tissue liquid extraction. The technique described in this study represents an important advance which broadens the application of SPME in vivo sampling technology.