The influence of circadian rhythms on pre- and post-prandial metabolism in the snake Lamprophis fuliginosus.

Measuring standard metabolic rate (SMR) and specific dynamic action (SDA) has yielded insight into patterns of energy expenditure in snakes, but less emphasis has been placed on identifying metabolic variation and associated energy cost of circadian rhythms. To estimate SMR, SDA, and identify metabolic variation associated with circadian cycles in nocturnally active African house snakes (Lamprophis fuliginosus), we measured oxygen consumption rates (VO2) at frequent intervals before and during digestion of meals equaling 10%, 20% and 30% of their body mass. Circadian rhythms in metabolism were perceptible in the VO2 data during fasting and after the initial stages of digestion. We estimated SMR of L. fuliginosus (mean mass=16.7+/-0.3 g) to be 0.68+/-0.02 (+/-SEM) mL O2/h at 25 degrees C. Twenty-four hours after eating, VO2 peaked at 3.2-5.3 times SMR. During digestion of meals equaling 10-30% of their body mass, the volume of oxygen consumed ranged from 109 to 119 mL O2 for SMR, whereas extra oxygen consumed for digestion and assimilation ranged from 68 to 256 mL O2 (equivalent to 14.5-17.0% of ingested energy). The oxygen consumed due to the rise in metabolism during the active phase of the daily cycle ranged from 55 to 66 mL O2 during digestion. Peak VO2, digestive scope, and SDA increased with increasing meal size. Comparisons of our estimates to estimates derived from methods used in previous investigations resulted in wide variance of metabolic variables (up to 39%), likely due to the influence of circadian rhythms and activity on the selection of baseline metabolism. We suggest frequent VO2 measurements over multiple days, coupled with mathematical methods that reduce the influence of undesired sources of VO2 variation (e.g., activity, circadian cycles) are needed to reliably assess SMR and SDA in animals exhibiting strong circadian cycles.

Responses of benthic fish exposed to contaminants in outdoor microcosms--examining the ecological relevance of previous laboratory toxicity tests.

Previous laboratory studies indicate that coal combustion wastes (a mixture composed of fly ash and other lower volume wastes such as bottom ash; hereafter collectively referred to as ash) adversely affect the health of benthic fish (Erimyzon sucetta; lake chubsucker), but fish in these studies were provided with ample uncontaminated food resources. Because aquatic disposal of ash can also adversely affect food resources for benthic fish, we hypothesized that changes in resources might exacerbate the effects of ash on fish observed in laboratory studies. We exposed juvenile E. sucetta in outdoor microcosms to water, sediment, and benthic resources from an ash-contaminated site or a reference site for 45 days and compared our findings to previous laboratory studies. Benthic invertebrate biomass was nearly three times greater in controls compared to ash microcosms. Total organic content of control sediment (41%) was also greater than in ash sediments (17%), suggesting that additional benthic resources may have also been limited in ash microcosms. Benthic invertebrates isolated from the ash microcosms had trace element concentrations (As, Cd, Co, Cr, Cs, Se, Sr, and V) up to 18 times higher than in weathered ash used in laboratory studies. The concentrations of trace elements accumulated by fish reflected the high dietary concentrations encountered in the ash microcosms and were associated with reduced growth (final mass = 0.07 g) and survival (25%) compared to controls (0.37 g and 67%, respectively). Accumulation of trace elements, as well as reductions in growth and survival, were more pronounced than in previous laboratory studies, suggesting that resource conditions may be important in mediating ash toxicity. Taken together, our studies suggest that ash discharge into aquatic systems is a more serious threat to the health of benthic fish than previously predicted based upon laboratory toxicity tests.

Altered swimming performance of a benthic fish (Erimyzon sucetta) exposed to contaminated sediments.

Numerous field surveys suggest that coal combustion wastes (ash) adversely affect fish populations, but few controlled laboratory studies have evaluated the responses of individual fish to ash exposure. Available information suggests that sublethal effects of ash, including decreased growth rates and reduced fecundity, may be important manifestations of ash toxicity. Here, we hypothesized that ash may also alter swimming performance, which could ultimately have important implications for the autecology of affected species. To test this hypothesis, we measured sprint speed and critical swimming speed (U(crit)) of juvenile lake chubsuckers (Erimyzon sucetta) exposed to ash in the laboratory. Fish exposed to ash for 90-100 days accumulated significant concentrations of As, Se, Sr, and V; exhibited severe fin erosion; and had reduced sprint speed and U(crit). Compared to controls, sprint speed of ash-exposed fish was reduced by 30% at 5 cm and the percent reduction was further reduced to 104% at 20 cm. Critical swimming speed was approximately 50% lower in fish exposed to ash compared to controls. Additionally, the typical positive relationship between standard length and U(crit) was absent in fish exposed to ash. Because reductions in swimming performance could not be attributed to pollutant-induced differences in body condition or fin morphology, we hypothesize that physiological disruptions (e.g., increased energy demands, decreased oxygen uptake or transport) are responsible for the observed effects and warrant further attention.

Effects of food ration on survival and sublethal responses of lake chubsuckers (Erimyzon sucetta) exposed to coal combustion wastes.

Study organisms in chronic toxicological bioassays are often provided with excessive resources to remove food limitations as a confounding experimental variable. Under more ecologically realistic situations, resources are often less abundant and such restrictions may alter the responses of organisms to environmental contaminants. Here, we investigated the interaction between resource level and sediment toxicity in the lake chubsucker, Erimyzon sucetta. For 78 days we fed fish one of three ration levels (1X, 2X, 4X; uncontaminated food) that was grazed directly from either clean sand or coal ash-contaminated sediments. Despite provision of uncontaminated food, fish exposed to the contaminated sediments accumulated significant whole body concentrations of As, Se, Sr, and V. Food ration affected the pattern of Se accumulation, with lowest concentrations accumulated by fish supplied with the lowest rations (1X). Paradoxically, fish in the 1X-ash treatment were most adversely effected by ash-exposure, despite having Se burdens much lower than fish in the 2X- and 4X-ash treatments. Fish in the 1X-ash treatment exhibited higher mortality, lower proportional growth, and increased incidence of fin erosion compared to fish provided with higher rations. Such results may, in part, be explained by the apparent inability of fish with reduced rations to maintain positive energy balance, as evidenced by their higher standard metabolic rates compared to control fish fed similar rations. Our results underscore the importance of considering resource quantity and nutritional factors in chronic bioassays in order to draw more ecologically realistic conclusions about contaminant effects.

Nondestructive indices of trace element exposure in squamate reptiles.

Compared with birds, mammals, fish, and even amphibians, very little is known about the effects of contaminants on reptiles. Recent evidence that many reptile populations may be declining has stimulated demand for toxicological studies of reptiles as well as development of nondestructive sampling techniques useful for assessing and monitoring contaminant exposure. The current study experimentally evaluated the utility of shed skins, tail clips, and blood samples as nondestructive indices of trace element exposure in banded water snakes, Nerodia fasciata. For 13.5 months, snakes were either fed fish from a coal ash-contaminated site or uncontaminated food from a reference site. Snakes fed contaminated prey accumulated As, Cd, Se, Sr, and V in various organs (i.e. liver, kidney, and/or gonads). Moreover, non-parametric discriminant function analysis revealed that snakes could be placed in two groups that reliably reflected their experimental diet based upon Se, Sr, and As concentrations in tail clips, blood, and/or shed skins. We suggest that nondestructive sampling techniques, particularly analyses of blood and tail clips, may be easily applied in evaluations of contaminant exposure in the field and laboratory and may prevent excessive destructive sampling of potentially threatened reptile species.

Detrimental effects associated with trace element uptake in lake chubsuckers (Erimyzon sucetta) exposed to polluted sediments.

Lake chubsuckers (Erimyzon sucetta) were exposed to coal ash-polluted sediments under conservative experimental conditions (filtered artificial soft water and abundant uncontaminated food). After 4 months of exposure, fish grazing the polluted sediments had significantly elevated body burdens of Se, Sr, and V. Selenium levels were particularly elevated, reaching mean whole body concentrations of 5.6 microg/g dry mass by the end of experimental manipulations. Twenty-five percent of fish exposed to pollutants died during the study. All surviving fish exposed to ash exhibited substantial decreases in growth and severe fin erosion. Total nonpolar lipids were two times higher in fish from the control treatment, but percent lipid did not differ between treatments. Because fish were presented with the same amount of food during the study, it appears fish exposed to ash utilized more energy for daily activities and/or were less efficient at converting available energy to tissues for growth and storage. The results were particularly interesting because we were unable to detect differences in standard metabolic rate (SMR) of fish between treatments. Increased energy expenditures not detectable in estimates of maintenance based on SMR, such as costs of digestion or activity, may have contributed to decreased energetic efficiency. Our findings corroborate previous studies which have documented the toxicity of ash-derived pollutants in fish.