Search

It is increasingly clear that the public, native tribes, and governmental agencies are interested in assessing the well-being of natural resources and ecosystems. This may take the form of understanding species presence, monitoring population status and trends, measuring behavior, or quantifying physiology, biological stresses, or chemical/radiological exposure through biomarkers. Often there is a separation between understanding the biological aspects of species well-being and assessing exposure to contaminants. In this paper we examine the applicability of using scuba sampling aimed primarily at specimen collection for radionuclide analysis to assess species presence/absence and to compare among sampling sites and depths. We were especially interested in whether dive transects could provide information on species presence and potential exposure to environmental contaminants. In June/July 2004 we sampled at 49 depth stations along 19 transects at Amchitka and Kiska Islands in the western Aleutian Islands in the Northern Pacific/Bering Sea region. Amchitka Island, a former World War II U.S. Navy base, was the site of three underground nuclear test shots from 1965 to 1971. Four to six transects were established at three Amchitka sites and two Kiska Sites, and 2 to 4 stations were sampled on each transect. Bottom conditions, weather and currents prevented a complete sampling of all stations. There were interspecific differences in the percent of stations where biota were found and collected, in their occurrence near the three test shots on Amchitka, and in the depth where they were found. There were no significant differences between Amchitka and Kiska Island in the percent of stations where species were found. These data suggest that information gathered incidentally to the collection of specimens for chemical/radiological analysis can prove useful for understanding the presence of benthic organisms along particular transects, at given depths, and at different geographical locations. This information also provides a baseline for the range of organisms that could be exposed to future physical or chemical/radiological stressors. The data are useful for developing future biomonitoring plans to assess biological well-being and chemical/radiological exposure only if they are published and available to the public, public policy makers, and managers. Just as it is critical to select endpoints and bioindicators that are of interest for assessing both human and ecological health, specimens should be collected using a protocol that is useful for both chemical/radiological analysis and biological information.

Recreational and subsistence fishing plays a major role in the lives of many people, although most Americans obtain their fish from supermarkets or other commercial sources. Fish consumption has generally increased in recent years, largely because of the nutritional benefits. Recent concerns about contaminants in fish have prompted federal and state agencies to analyze fish (especially freshwater fish targeted by recreational anglers) for contaminants, such as mercury and polychlorinated biphenyls (PCBs), and to issue fish consumption advisories to help reduce the public health risks, where warranted. Scientists engaged in environmental sampling collect fish by a variety of means, and analyze the contaminants in those fish. Risk assessors use these levels as the basis for their advisories. Two assumptions of this methodology are that scientists collect the same size (and types) of fish that fishermen catch, and that, for some contaminants (such as methylmercury and PCBs), levels increase with the size and age of the fish. While many studies demonstrate a positive relationship between size and mercury levels in a wide range of different species of fish, the assumption that scientists collect the same size fish as fishermen has not been examined. The assumption that scientists collect the same size fish as those caught (and eaten) by recreationalists or subsistence fishermen is extremely important because contaminant levels are different in different size fish. In this article, we test the null hypothesis that there are no differences in the sizes of fish collected by Aleut fishermen, scientists (including divers), and commercial trawlers in the Bering Sea from Adak to Kiska. Aleut fishermen caught fish using rod-and-reel (fishing rods, hook, and fresh bait) from boats, as they would in their Aleutian villages. The scientists collected fish using rod-and-reel, as well as by scuba divers using spears up to 90 ft depths. A fisheries biologist collected fish from a research/commercial trawler operated under charter to the National Oceanographic and Atmospheric Administration (NOAA). The fish selected for sampling, including those caught commercially in the Bering Sea, represented different trophic levels, and are species regularly caught by Aleuts while fishing near their villages. Not all fish were caught by all three groups. There were no significant differences in length and weight for five species of fish caught by Aleuts, scientists, and fisheries trawls, and for an additional 3 species caught only by the Aleut and scientist teams. There were small, but significant, differences in the sizes of rock greenling (Hexagrammos lagocephalus) and red Irish lord (Hemilepidotus hemilepidotus) caught by the scientist and Aleut fishermen. No scientists caught rock greenling using poles; those speared by the divers were significantly smaller than those caught by the Aleuts. Further, there were no differences in the percent of males in the samples as a function of fishing method or type of fishermen, except for rockfish and red Irish lord. These data suggest that if scientists collect fish in the same manner as subsistence fishermen (in this case, using fishing rods from boats), they can collect the same-sized fish. The implications for exposure and risk assessment are that scientists should either engage subsistence and recreational fishermen to collect fish for analysis, or mimic their fishing methods to ensure that the fish collected are similar in size and weight to those being caught and consumed by these groups. Further, total length, standard length, and weight were highly correlated for all species of fish, suggesting that risk assessors could rely on recreational and commercial fishermen to measure total lengths for the purpose of correlating mercury levels with known size/mercury level relationships. Our data generally demonstrate that the scientists and trawlers can collect the same size fish as those caught by Aleuts, making contaminant analysis, and subsequent contaminant analysis, representative of the risks to fish consumers.

The "peroxidase" activity of the copper-zinc superoxide dismutase is a poorly sustained activity because of the competing inactivation of the enzyme. New evidence suggests that the bound oxidant may be partitioning between oxidizing the enzyme or oxidizing small anions. At constant peroxide, nitrite and azide only partially protect the enzyme (50%) against loss of copper(I) and inactivation up to one anion per copper. Beyond that level, there is no further protection. Bicarbonate ion also protects, but larger amounts are required. These data suggest that there is significant oxidation of the enzyme even in the presence of the small anions and therefore the formation of the bound oxidant cannot be sustained in a true catalytic process., (Copyright 2000 Academic Press.)

N,N'-Diethyldithiocarbamate has been shown to be an analytical stain for copper in native polyacrylamide gels of the copper-zinc superoxide dismutase from purified preparations as well as from crude red cell extracts, i.e., lysates from which hemoglobin has been removed (Jewett, S. L., and Rocklin, A. M. (1994) Anal. Biochem. 217, 236-240). Applying this methodology, it was found that the relative amounts of copper-containing forms of copper-zinc superoxide dismutase (EC 1.15.1.1) from bovine red cell extracts did not change significantly with either the age or with hydrogen peroxide treatment of the red cells. Furthermore, no significant changes were seen in the specific activity of the dismutase in either type of experiment. These observations for both types of experiments are contrary to what was expected from similar studies reported in the literature. However, discrepancies may be accounted for by hemoglobin interference in indirect dismutase assays of the previous work. In the case of the peroxide treatment of red cells, however, there is an additional factor in that the dismutase is protected from peroxide-mediated changes in copper content and heterogeneity by the hemoglobin present. This protection was demonstrated in in vitro experiments using only a 24-fold excess of hemoglobin over the dismutase.

N,N'-Diethyldithiocarbamate reacts with copper in the copper-zinc superoxide dismutase (EC 1.15.1.1) in polyacrylamide gels to form stable yellow-brown bands that are quantifiable at 448 nm. This method of examining superoxide dismutase has been applied to crude extracts of the enzyme obtained from red cell lysates from which hemoglobin has been removed by chloroform-ethanol precipitation. This treatment did not affect the activity and heterogeneity of purified dismutase added to lysates and recovered by the same method. The bands that develop in the dithiocarbamate-stained gels of the extracts correspond exactly to the bands of dismutase activity obtained with a positive activity stain using dianisidine, indicating that the dismutase is the only copper protein that gives rise to these bands. The amount of superoxide dismutase in the bands, determined by comparing the areas under unknown peaks to areas obtained with a standard dismutase sample, agrees with the amount predicted from indirect superoxide dismutase activity measurements. Any color in gels due to trace hemoglobin or hemoglobin degradation products is bleached overnight during staining with the dithiocarbamate.

Although numerous reviews and articles have been written about indirect superoxide dismutase assays, little has been reported about the variation of the amount of superoxide dismutase giving 50% inhibition (SOD50), defined as one unit of dismutase activity, with variations in assay conditions. This report indicates that there is a linear dependence of the SOD50 on the magnitude of the uninhibited rate of oxidation of the organic substance for two indirect assays, one involving epinephrine autoxidation and the other NADH oxidation. For one indirect assay involving an acceleration of the oxidation rate of an organic substrate, the amount of superoxide dismutase giving a doubling of the oxidation rate, defined as one unit of dismutase activity, also shows this linear dependence on the autoxidation rate of the organic substance. The significance of this dependence and its implications for experimenters using these assays is discussed.