This study consisted of a site characterization followed by a biomonitoring study utilizing the zebra mussel, <i>Dreissena polymorpha</i>, at the Times Beach Confined Disposal Facility (CDF), located in Buffalo, New York. Concentrations of the selected contaminants (polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), and the following metals: arsenic (As), chromium (Cr), barium (Ba), mercury (Hg), cadmium (Cd), lead (Pb), selenium (Se) and silver (Ag), were at or below detection limits in the water column. In the sediment toxicant concentrations were as high as 549 mg/kg for total PAHs, 9 mg/kg for PCB Aroclor 1248, and 54, 99, 6, 355, 637, and 16 mg/kg for the metals: As, Ba, Cd, Cr, Pb, and Hg respectively. To predict contaminant bioavailability, elutriate and whole sediment toxicity tests were performed utilizing the cladoceran, <i>Daphnia magna</i>. The whole sediment tests showed a significant impact. Control survival was 84%, while the sediment treatment survival range was 1-7%. Mean control reproduction was 86.8 neonates, whereas treatment group reproduction ranged from 1.4 to 9.0. Zebra mussels, placed both in the water column (Upper) and at the sediment level (Lower), survived the 34-day exposure. Contaminants which significantly accumulated in zebra mussel tissue during the exposure period (mg/kg) were total PAHs (6.58), fluoranthene (1.23), pyrene (1.08), chrysene (0.98), benzo(a)anthracene (0.60), PCB Aroclor 1248 (1.64), As (0.97), Cr (2.87), and Ba (7.00). The accumulation of benzo(a)anthracene was statistically higher in the Upper mussels; however, this did not occur for any other toxicant. Accumulation of these contaminants in zebra mussel tissue represent a potential hazard to organisms (ie. fish and birds) which feed on them. / Master of Science

The objective of this study was to evaluate the impact of extreme acidic and alkaline pH excursions of fish populations in streams receiving fly ash settling basin effluent. Fish communities were sampled above and below the effluent outfall in a mountain stream, Adair Run, adjacent to the coal-fired Glen Lyn Power Plant in southwestern Virginia. This effluent, which exhibited a pH increase of up to pH 9.3 as the basin nearly filled, contributed to a downstream decline in species diversity and an increased dominance of the stoneroller (Campostoma anomalum), which displaced more sensitive species. Laboratory avoidance studies, utilizing a steep-gradient trough apparatus, were used to investigate the potential role of avoidance behavior that may alter fish distributions in acidic and alkaline waters. Stonerollers, rainbow trout (Salmo gairdneri), golden shiners (Notemigonus crysoleucas) and spotfin shiners (Notropis spilopterus) exhibited first significant avoidance responses to continuously decreasing pH conditions at pH 6.0-7.0, well above the 96-hr LC50 values of 4.05 and 4.24 for trout and golden shiners, respectively. When the pH was gradually increased, avoidance thresholds of pH 9.5- 11.0 were observed, beyond the 96-hr LC50 values of 9.13 and 8.86 for trout and golden shiners. Single acute alkaline exposures initially yielded more rapid avoidance responses, with acclimation to alkaline conditions frequently occurring within 30 minutes. Gill histological alterations, as observed by transmission electron microscopy, were detected following exposure to extreme pH excursions, with more rapid changes occurring with alkaline exposures. Gill tissue showed marked recovery within 8-14 days. / Master of Science

The effects of sublethal copper on metabolism were in vestigated in bluegill (Lepomis macrochirus) by measuring whole body oxygen consumption in fish exposed to sublethal copper alone and in conjunction with a temperature increase. In vitro oxygen consumptions of liver, brain, and gill were also measured under these two conditions, as was the accumulation of copper in these tissues. In addition, the concentration of copper in bile was measured. Copper was found to decrease whole body oxygen consumption in animals exposed to copper alone, although the oxygen consumptions of tissues were not significantly altered. This indicates that copper is acting to decrease VO₂ at a higher level of integration than the individual tissues. In animals subjected to an increase in temperature as well as sublethal copper, oxygen consumption was higher than controls five days after the temperature was increased, indicating a delay in temperature acclimation. This increase was reflected in higher in vitro oxygen consumption in the liver and gill indicating that sublethal copper delays temperature acclimation by acting directly on the tissues. Tissue copper accumulation was seen first in the gills followed by accumulation in the liver. Copper was not found to accumulate in the brain. Increased copper levels were found in the bile at all tested exposure times. A discussion of the ecological implications of these findings is included. / M. S.

Changes in heart rate, ventilatory activity and oxygen consumption were determined in trout (Salmo gairdneri) and brown bullhead catfish (Ictalurus nebulosus) during exposure to a steadily increasing concentration of waterborne cyanide selected to produce death in 8-9 hours for each species. The lethal cyanide concentration for the bullheads was an order of magnitude higher than for trout. Trout developed an immediate and gradually increasing bradycardia throughout the exposure period. Cyanide produced tachycardia in the bullhead followed by a gradual onset of bradycardia as the concentration of cyanide was raised. Pericardial injection of atropine (a muscarinic cholinergic antagonist) indicated that bradycardia in the trout was due initially to increased vagal tone but later due to the direct effect of cyanide on the heart. Hyperventilation in the trout persisted throughout the exposure period, although the rate and amplitude fluctuated and was variable between individual fish. During the last hour of exposure (highest cyanide concentration), ventilation was characterized by rapid, shallow breaths with a sudden respiratory arrest. The bullheads showed hyperventilation during the first 3 hours of exposure followed by a gradual, linear drop in ventilation rate and amplitude until death occurred. Cardiac and ventilatory responses in both species were attributed to stimulation of central and peripheral chemoreceptors by cyanide. Evidence is presented which suggests the initial response in the bullheads was due, at least in part, to gustatory stimulation by the cyanide. Oxygen consumption of the trout remained above pre-exposure levels for the majority of the test period. Oxygen consumption in the bullhead paralleled the changes in heart and ventilatory rates. Whole-body lactate and pyruvate levels of fingerlings of both species during cyanide exposure were measured to estimate the extent of anaerobiosis. Whole-body lactate levels were greater in the bullheads than the trout, indicating a higher capacity for anaerobiosis, probably due to a greater fuel supply. Whole-body pyruvate levels in both species did not change from control levels until the last hour of cyanide exposure. / M.S.

Studies were conducted to evaluate the effect of pre-irrigation media moisture deficit, irrigation application rate, and intermittent irrigation on irrigation efficiency [(amount applied - amount leached) amount applied' 100] of spray stake-irrigated, container-grown plants. In the first experiment, pine bark-filled containers were irrigated to replace moisture deficits of 600, 1200, or 1800 ml. Deficits were returned in single, continuous applications at application rates of 148, 220, and 270 ml/min. Application rate did not affect irrigation efficiency. Efficiency decreased with increased medium moisture deficit. In the second experiment, containers, at 600 ml media moisture deficits, were irrigated with 400 or 600 ml (65% and 100% water replacement, respectively). Irrigation volumes were returned in a single, continuous application or in 100ml applications with 30 min intervals between irrigations (intermittent). Irrigation efficiency was greater with intermittent irrigation, 95 % and 84 % for 400 and 600 ml replacement, respectively, than with continuous irrigation, 84% and 67% for 400 and 600 ml replacement, respectively. When applied intermittently, most water loss from containers occurred after 400 ml were applied. In the third experiment, containers were irrigated with 600 ml (100% water replacement) in 50, 100, or 150 ml applications with 20, 40, or 60 min intelVals between applications. A control was included in which 600 ml was applied in a single, continuous application. Irrigation efficiency increased with decreasing application volume and increasing length of interval between applications. The greatest efficiency (86%) was produced with an irrigation regimen of 50 ml applications with at least 40 min between applications. The control treatment (continuous) produced an irrigation efficiency of 62%. When applied intermittently, most water loss from containers occurred after a total of 300 ml were applied. / Master of Science

The purpose of this study was to compare the simultaneous effects of the water soluble fraction (WSF) of JP-4, a common military and civilian jet fuel, on survival, ventilatory rate, preference-avoidance behavior, and blood enzymes and ions of the bluegill sunfish (Lepomis rnacrochirus) to determine possible interrelationships and which procedures might be most descriptive of sublethal stress. The dynamic 96-h LCSO for WSF JP-4 was 26.2% (percent of the maximum soluble amount of JP-4). A concentration of 5.1% WSF JP-4 caused a detectable shift in ventilatory functions (rate and amplitude); this concentration was equivalent to the 96-h LCOl. Fish did not display a strong preference-avoidance reaction when exposed to WSF JP-4. Some avoidance occurred at 3.5% and 4.9% WSF JP-4. At concentrations near the 96-h LCSO fish appeared to lose their ability to detect WSF JP-4, indicating potential for fish not to avoid lethal levels of WSF JP-4. Few significant changes in whole and serum blood parameters were measured in fish exposed to 13% WSF JP-4; at near lethal concentrations changes were primarily attributed to osmoregulatory failure. Therefore such changes in blood parameters would not be useful to validate or confirm exposure to WSF JP-4. There were few significant changes in gill and liver histology of fish exposed to WSF JP-4 other than an increase in size or number of lipid vacuoles in the liver of exposed fish. / Ph. D.

Chlorella vulgaris and Chlorella pyrenoidosa were compared relative to their abilities to grow at osmotic potentials of -0.1, -0.5, -1.0, -1.5, -2.0 MPa, [polyethylene glycol(PEG)-induced], and for osmotically-induced changes in lipid composition. C. vulgaris growth was inhibited as osmotic potentials decreased, while C. pyrenoidosa growth was moderately inhibited at -2.0 hPa. C. vulgaris produced increasing concentrations of triglycerides and sterol esters and decreasing levels of polar lipids and sterols as osmotic concentrations increased. Polar lipids, triglycerides, and sterols declined in C. pyrenoidosa while steryl esters remained constant. Ratios of free sterols to polar lipids were 10-fold greater in C. pyrenoidosa and were unaffected by reduced osmotic potentials. In C. vulgaris the sterol to polar lipid ratio declined. Decreasing osmotic potentials in a continuous culture of C. vulgaris, lowered cell lipid concentration, and had no effect on chlorophyll concentrations. The greatest decrease occurred as the osmotic potential decreased from -0.1 to -0.5 HPa. Decreasing osmotic potential caused the phospholipid concentrations to decline. Saturation of triglycerides and free fatty acids increased and decreased, respectively, while polar lipids remained fairly constant. However, the sterol to phospholipid ratio increased as the osmotic potential was lowered. BAS 13-338 (4-chloro-5-(dimethylamino)-2-phenyl-5- 3(2H)pyridazinone) had no effect on C. vulgaris resistance to osmotic stress, but caused growth inhibition as concentrations increased. However, BAS 13-338 was effective in decreasing growth inhibition of C. vulgaris grown in inhibitory levels of ethanol. BAS 13-338 had differing effects on the lipid composition of C. vulgaris when grown in PEG at an osmotic potential of -1.5 MPa compared to -0.1 MPa with 0.33% ethanol. The greatest effects were observed in the ethanol treatments where the qualitative composition of precursor sterols increased as the level of BAS 13-338 increased. This investigation confirmed the important role of lipids in responding to environmental stress through observations of lipid responses to osmotic stress and by manipulation of lipid concentrations using BAS 13-338. Resistance to ethanol inhibition but not osmotic inhibition was achieved in the investigation. / Ph. D.

This research describes aquatic macroinvertebrate community responses to heavy metals (copper, zinc) in experimental streams and at metal-impacted sites in the field. Experiments employed substrate-filled trays which were colonized in the field and then transferred to laboratory or outdoor streams. Laboratory experiments conducted over three seasons showed that acute (96 h) exposure to copper (Cu) at 15-32 μg Cu/L significantly reduced macroinvertebrate abundance and number of taxa during each season. Owing to differences in sensitivity among taxa, the percent composition of dominant groups varied between control and dosed streams. Mayflies were quite sensitive to Cu, particularly during the summer when water temperatures were higher. Community responses to Cu and Zn in outdoor experimental streams were similar to those observed at metal-impacted sites in the field. Control streams and field reference Stations were dominated by mayflies and Tanytarsini chironomids. In contrast, treated Streams and impacted field sites were dominated by net-Spinning caddisflies (Hydropsychidae) and Orthocladiini chironomids. The similarity of these experimental results to those observed in the field suggest that macroinvertebrate community responses to heavy metals are highly predictable. Responses of these communities to Cu were greatly influenced by water quality. Effects were more severe in New River Streams, where water hardness and alkalinity were low, compared to Clinch River Streams, where hardness and alkalinity were higher. In soft water Streams, abundance was reduced by 84% after 10 d exposure to Cu (measured concentration = 13 μg/L). In contrast, abundance was reduced by only 45% in hard water Streams after 10 d at Similar Cu levels. These results demonstrate the importance of accounting for water quality characteristics of receiving systems when establishing site-specific criteria for metals. Chronic exposure (14 d) to sublethal levels of Cu (< 6 μg/L) increased Vulnerability of caddisflies (Hydropsyche morosa and Chimarra sp.) to predation by the Stonefly, Paragnetina fumosa. Caddisflies were also the major component of stonefly diets and were consumed Significantly more frequently in dosed Streams than controls. These results demonstrate that single Species bioassays were inadequate for predicting effects of toxicants on community level processes. / Ph. D.

Effects of hexavalent chromium on the life cycle of Chironomus riparius were examined in the laboratory, using sublethal (0.5 mg/1) and 40% lethal (2.0 mg/1) concentrations. Chironomus egg masses were introduced into both control and chromium-treated 21 1 aquaria. Larvae were subsampled at 5-day intervals, measured for length, and weighed to generate growth curves. Emerging adults were allowed to mate and oviposit. Twenty female pupae were collected from control and treatment (0.5 mg/1) populations within 12 hours of pupation. Their bodies were analyzed for lipid content by calculating loss in dry weight after a chloroform-methanol extraction. Chromium body burdens were measured in both larvae and adults from treatment (2.0 mg/1) and control populations. Effects of chromium on hatching rates of egg masses were studied. Growth was stimulated at both lethal and sublethal concentrations of chromium. Lipid content of female pupae was significantly higher in treatment populations than in controls. Both stimulated growth and increased lipid storage may be the results of metal activation of enzymes involved in energy processing and storage in the insect. Reproductive success, as measured by numbers of offspring per adult female, declined with increasing chromium concentrations. Chromium concentrations in adult males and larvae were significantly greater for treatment populations. Metal body burdens may have become concentrated in eggs or sperm of Chironomus and caused the observed decrease in reproductive success. No direct toxicity of chromium on egg masses was observed at concentrations up to 5 mg/1. / M.S.

Chan Kwan-ling. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2005. / Includes bibliographical references (leaves 91-105). / Abstracts in English and Chinese. / Abstract --- p.i / Acknowledgements --- p.v / Contents --- p.vi / List of figures and tables --- p.ix / Chapter Chapter 1 --- Introduction / Chapter 1.1 --- Literature review --- p.1 / Chapter 1.1.1 --- The effect of pollutants on the genetic materials of aquatic organisms --- p.1 / Chapter 1.1.1.1 --- Response of individual to genotoxicants --- p.1 / Chapter 1.1.1.2 --- Effects of genotoxicants on population structure --- p.3 / Chapter 1.1.2 --- Application of genetic markers in monitoring water pollution --- p.3 / Chapter 1.1.2.1 --- DNA adduct --- p.4 / Chapter 1.1.2.2 --- Sister chromatid exchange (SCE) test --- p.5 / Chapter 1.1.2.3 --- Micronucleus --- p.6 / Chapter 1.1.2.4 --- DNA strand breaks --- p.7 / Chapter 1.1.3 --- Single-cell gel electrophoresis (comet) assay --- p.9 / Chapter 1.1.4 --- Test organisms for comet assay --- p.12 / Chapter 1.2 --- Objective of the present study --- p.13 / Chapter Chapter 2 --- Genotoxicity of pollutants on Hyale crassicornis / Chapter 2.1 --- Introduction --- p.22 / Chapter 2.2 --- Materials and methods --- p.24 / Chapter 2.2.1 --- Sampling of amphipods --- p.24 / Chapter 2.2.2 --- Acclimation --- p.24 / Chapter 2.2.3 --- Acute toxicity test --- p.26 / Chapter 2.2.4 --- The effect of test duration on DNA damage --- p.27 / Chapter 2.2.5 --- Effect of toxicants on DNA damage --- p.28 / Chapter 2.2.6 --- Comet assay --- p.29 / Chapter 2.2.7 --- Chemicals --- p.34 / Chapter 2.2.8 --- Data analysis --- p.34 / Chapter 2.3 --- Results --- p.34 / Chapter 2.4 --- Discussion --- p.47 / Chapter Chapter 3 --- Genotoxicity of hydrogen peroxide on different tissue types of Metapenaeus ensis / Chapter 3.1 --- Introduction --- p.54 / Chapter 3.2 --- Materials and Methods --- p.57 / Chapter 3.2.1 --- Collection and acclimation of shrimps --- p.57 / Chapter 3.2.2 --- Incubation --- p.59 / Chapter 3.2.3 --- Comet Assay --- p.60 / Chapter 3.2.4 --- Chemicals --- p.61 / Chapter 3.2.5 --- Data analysis --- p.61 / Chapter 3.3 --- Results --- p.61 / Chapter 3.4 --- Discussion --- p.67 / Chapter Chapter 4 --- Genotoxicity of wastewater on Hyale crassicornis / Chapter 4.1 --- Introduction --- p.71 / Chapter 4.2 --- Materials and Methods --- p.72 / Chapter 4.2.1 --- Collection of wastewater samples --- p.72 / Chapter 4.2.2 --- Metal content analysis --- p.73 / Chapter 4.2.3 --- Genotoxic effect of wastewater samples on Hyale crassicornis --- p.74 / Chapter 4.2.4 --- Chemicals --- p.76 / Chapter 4.2.5 --- Statistical analysis --- p.77 / Chapter 4.3 --- Results --- p.77 / Chapter 4.3.1 --- Metals content in water samples --- p.77 / Chapter 4.3.2 --- DNA damage --- p.79 / Chapter 4.4 --- Discussion --- p.79 / Chapter Chapter 5 --- Conclusions --- p.89 / References --- p.91