Biochemical and Physiological Responses in Atlantic Salmon (Salmo salar) Following Dietary Exposure to Copper and Cadmium

Three experiments were conducted with Atlantic salmon (Salmo salar) to assess the effects of dietary exposure to copper and cadmium. The results presented here provide an overview, details of each experiment will be published in full elsewhere. In the first experiment, salmon parr exposed for four weeks to 35 and 700 mg Cu kg⁻¹ diet had significantly elevated intestinal copper concentrations, cell proliferation (PCNA) and apoptosis rates compared to control fish. No differences were observed in gill or plasma copper concentrations among the groups. In contrast to the controls, the Cu exposed groups did not grow significantly during the exposure period. The second experiment (three months exposure) was conducted to assess the effects of dietary copper (control, 35, 500, 700, 900 or 1750 mg Cu kg⁻¹ diet) on growth and feed utilization in salmon fingerlings. Growth was significantly reduced after three months exposure to dietary Cu concentrations above 500 mg kg⁻¹. Similarly, copper body burdens were significantly higher in fish exposed to elevated dietary copper concentrations (above 35 mg Cu kg⁻¹ diet). In the third experiment, salmon parr were exposed to one of six dietary cadmium concentrations (0, 0.5, 5, 25, 125 or 250 mg Cd kg⁻¹ diet) for four months. Cadmium accumulated in the liver>intestine>gills of exposed fish. Rates of apoptosis and cell proliferation in the intestine increased following exposure to dietary cadmium. Exposure to elevated concentrations of dietary cadmium had no effect on growth in salmon parr. Results from these studies indicate that cellular biomarkers have potential as early warning signs of negative effects on the overall fitness of an organism.     

Chronic effects of tributyltin on the caprellid amphipod Caprella danilevskii

In order to examine the biological effects by TBT exposure, experiments involving the exposure of five levels of TBT concentrations (0, 10, 100, 1000 and 10,000 ngl(-1)) were conducted on the caprellid amphipod Caprella danilevskii, over a generation after hatching. Marked delays in growth and molting during the early developmental stage and maturation were found in both 100 and 1000 ng TBTCl l(-1) concentrations in spite of the sex. All specimens died in 10,000 ng TBTCl l(-1) within 4 days after hatching. Inhibition of maturation and reproduction such as delaying in the achievement of maturity and a decrease in the number of juveniles hatched was apparent in 10 and 100 ng TBTCl l(-1) concentrations. Furthermore, brood loss, and failure in egg formation and hatching were observed as the TBT concentration became higher. No significant changes in sex ratio were seen at any of the TBT exposure levels during hatching and maturation in the present study. This phenomenon was different from a previous study in which the proportion of females was increased with an increase in TBT concentrations in the embryonic exposure experiment. This suggests that sex disturbance is induced during the embryonic stage of the species. A drastic decrease in survival rate was observed at 10 ng TBTCl l(-1) (25%) which corresponds to the mean level in coastal waters. Therefore, the present encountering ambient TBT concentration may influence populations of C. danilevskii in the coastal environment.     

Development of a laboratory exposure system using marine fish to carry out realistic effect studies with produced water discharged from offshore oil production

A biotest system for environmentally realistic exposure of fish to produced water (PW) was developed and tested. Authentic PW was collected at an oil production platform in the North Sea and preserved by freezing in multiple aliquots a 25 L. After transport to the test laboratory onshore, daily PW aliquots were thawed, homogenised and administered to the test fish, Atlantic cod (Gadus morhua), in two diluted exposure concentrations, 0.1% and 0.5%, during a 15 d period, using a continuous flow-through exposure setup. Positive control groups were exposed to two crude oil treatments for comparison. Chemical analyses showed that alkylphenol (AP) and PAH concentrations in PW exposure waters were very low. Observations of significantly increased AP and PAH metabolite levels in PW exposed fish demonstrated the suitability of the biotest system for its use in biological exposure/effect studies of PW, and it also demonstrated the sensitivity of bile metabolites as PW exposure markers in fish. The relevance of the biotest system for PW effect studies and for validating modelled environmental risk estimates of PW dischargers from offshore oil production is discussed.     

Organochlorine contaminant exposure and associations with hematological and humoral immune functional assays with dam age as a factor in free-ranging northern fur seal pups (Callorhinus ursinus)

Potential effects of organochlorine contaminant (OC) exposure on humoral immune function and health of free-ranging northern fur seals were investigated. Forty-two "neonates" were captured for blood sample collection and re-sampled as "pups" 29-51 days later. OCs were extracted from whole blood to identify polychlorinated biphenyl congeners and chlorinated pesticides by high performance liquid chromatography. Humoral immune function was assessed by antibody responses to vaccination and immunoglobulin concentrations. Additional health status indicators included leukocyte counts and haptoglobin concentrations. Mean OC concentrations were higher in neonates than at recapture. Neonates of young dams had higher mean OC concentrations than neonates of older dams. A higher proportion of old dam's pups developed a twofold or greater increase in antigen-specific antibodies compared to young dam's pups. Higher OC exposure and poor immune responses in first-born pups may indicate a higher risk of secondary morbidity and mortality than pups of multiparous dams.     

Contamination and toxic effects of persistent endocrine disrupters in marine mammals and birds

In recent years, several species of marine mammals and birds have been affected by uncommon diseases and unusual mortalities. While several possible causative factors have been attributed for these events, a prominent suspect is exposure to man-made toxic contaminants. Particularly, some of these man-made chemicals can disrupt normal endocrine physiology in animals. At CMES, our studies focus on exposure and toxic effects of endocrine disrupting chemicals, particularly organochlorines, in higher trophic level wildlife. Endocrine disrupting chemicals, such as organochlorine insecticides, polychlorinated biphenyls, organotins etc. are found in tissues of a wide variety of wildlife. Extremely high concentrations have been found in animals afflicted with diseases and/or victims of mass mortalities. Elevated contamination by organochlorines has been found in open sea animals such as cetaceans and albatrosses, which seemed to be attributable to their low capacity to metabolize toxic persistent contaminants. Significant correlations between biochemical parameters (serum hormone concentrations and cytochrome P450 enzyme activities) and residues of endocrine disrupting chemicals were found in some species of marine animals, which indicates that these chemicals may impose toxic effects in animals even at the current levels of exposure. In general, water birds and marine mammals accumulated the dioxin-like compounds with much higher concentrations than humans, implying higher risk from exposure in wildlife. The future issues of endocrine disrupting chemicals in humans and wildlife will have to be focused in developing countries.     

Lysosomal and lipid-associated parameters in the livers of three species of arctic seabird chicks: species differences and relationships with contaminant levels

Lysosomal membrane stability, lipofuscin (LF), malondialdehyde (MDA), neutral lipid (NL) levels, as well as halogenated organic compounds (HOCs), Cr, Cd, Pb and Fe concentrations were analyzed in liver of black-legged kittiwake (BK), herring gull (HG), and northern fulmar (NF) chicks. There were significant species differences in the levels of NL, LF and lysosomal membrane stability. These parameters were not associated with the respective HOC concentrations. LF accumulation was associated with increasing Cr, Cd and Pb concentrations. HG presented the lowest lysosomal membrane stability and the highest.LF and NL levels, which indicated impaired lysosomes in HG compared to NF and BK. Lipid peroxidation was associated with HOC and Fe²⁺ levels. Specific HOCs showed positive and significant correlations with MDA levels in HG. The study indicates that contaminant exposure can affect lysosomal and lipid associated parameters in seabird chicks even at low exposure levels. These parameters may be suitable markers of contaminant induced stress in arctic seabirds.     

Effects of the herbicide diuron on the early life history stages of coral

The effects of the herbicide diuron on the early life history stages of broadcast spawning and brooding corals were examined in laboratory experiments. Fertilisation of Acropora millepora and Montipora aequituberculata oocytes were not inhibited at diuron concentrations of up to 1000 microg l(-1). Metamorphosis of symbiont-free A. millepora larvae was only significantly inhibited at 300 microg l(-1) diuron. Pocillopora damicornis larvae, which contain symbiotic dinoflagellates, were able to undergo metamorphosis after 24 h exposure to diuron at 1000 microg l(-1). Two-week old P. damicornis recruits on the other hand were as susceptible to diuron as adult colonies, with expulsion of symbiotic dinoflagellates (bleaching) evident at 10 microg l(-1) diuron after 96 h exposure. Reversible metamorphosis was observed at high diuron concentrations, with fully bleached polyps escaping from their skeletons. Pulse amplitude modulation (PAM) chlorophyll fluorescence techniques demonstrated a reduction in photosynthetic efficiency (Delta F/F(m)') in illuminated P. damicornis recruits after a 2 h exposure to 1 microg l(-1) diuron. The dark-adapted quantum yields (F(v)/F(m)) also declined, indicating chronic photoinhibition and damage to photosystem II.     

Toxicity of Distillery Effluent to the Cyprinid Weed Fish Rasbora daniconius (Ham).

The toxicity of distillery wastes to the fish Rasbora daniconius was tested by bioassay. Ten concentrations ranging from 1.0 to 100% were employed. It was found that the fish were quite tolerant at the 1% concentration for about 24 hours of exposure and survived even up to 96 hours. Mortality was noticed in concentrations from 1.5% onwards. In the concentrations of 3.0% and above there was a heavy mortality beginning at 15 hours of exposure. From the chemistry of the medium studied it was found that the toxicity was inversely proportional to pH but directly proportional to conductivity and bicarbonate alkalinity of the medium.     

Effects of Short‐term and Long‐term Aluminium Stress on Photosynthesis,Respiration, and Reproductive Capacity in a Unicellular Green Flagellate (Euglena gracilis)

Effects of short‐term (1 h exposure) and long‐term (7 d exposure) aluminium stress on photosynthesis and reproductive capacity have been studied in Euglena gracilis strain Z. Following concentrations of Al_tot (added as AlCl₃) were tested: 0.5 mg L^‐1, 1.0 mg L^‐1, 1.5 mg L^‐1, 2.5 mg L^‐1, 5.0 mg L^‐1, 7.5 mg L^‐1, 10.0 mg L^‐1, and 15.0 mg L^‐1 Al, respectively. Growth rates at different aluminium concentrations did not show significant differences, except at 15.0 mg L^‐1Al. Initial respiration was higher in long‐term than in the short‐term experiments. It is supposed that an energy‐dependent mechanism of excretion of aluminium ions has been active in the stressed cells. Consequently, the cells of E. gracilis after long‐term exposure to aluminium are believed to be more acclimatised to the aluminium stress. Photosynthetic efficiency (PE) has been negatively affected by aluminium in all experiments performed. Differences between control algae and those treated with aluminium were significant in all cases. PE in long‐term experiments was in general significantly higher at all concentrations of aluminium studied, compared to the short‐term experiments. The aluminium concentrations tested led only to a general decrease in PE while the level of decrease was not especially concentration‐dependent. In general, aluminium tolerance of E. gracilis can be estimated as low, especially by short‐term exposure. However, good acclimatisation capacity of this green flagellate to aluminium doses by long‐term exposure can be supposed.     

Evidence of polycyclic aromatic hydrocarbon (PAH) exposure in fish from the Antarctic peninsula

PAH exposure was assessed in fish from Arthur Harbor, Antarctica and sites remote from human activities by measuring the concentrations of polycyclic aromatic hydrocarbon (PAH) metabolites in bile and PAH in tissues and stomach contents. The highest concentrations of biliary metabolites and tissue PAH were present in fish captured near an Argentine supply ship (Bahia Paraiso) that ran aground in Arthur Harbor in 1989. The presence of biliary PAH metabolites in fish collected near Palmer Station, a small US scientific station in Arthur Harbor, suggests low level exposure to contaminants. However, PAH tissue concentrations in fish from Palmer Station were similar to those from remote sites. Both tissue PAH and metabolite concentrations were low in fish captured at sites remote from human activities. The confirmation of PAH metabolites in the bile of fish from Arthur Harbor is direct evidence that the presence of humans has resulted in Antarctic fish being exposed to petroleum-derived PAH.     

Exposure of the clam Tapes philippinarum to 4-nonylphenol: changes in anti-oxidant enzyme activities and re-burrowing capability

The effects of 4-nonylphenol on anti-oxidant enzyme (superoxide dismutase and catalase) activity in both gills and digestive gland and re-burrowing capability of the clam Tapes philippinarum were investigated after 7 days' exposure to various sublethal concentrations of nonylphenol (NP). NP caused a significant decrease in superoxide dismutase activity in both tissues from the lowest concentration tested, whereas no significant alteration with respect to controls was observed in catalase activity. NP effects on re-burrowing of clams were evaluated in two experiments. In the first, clams exposed to the highest NP concentrations showed a marked delay in re-burrowing 48 h after the end of exposure. Modifications in normal burrowing behaviour were also observed, suggesting that NP acts via narcosis and/or neurotoxic mechanisms. In the second experiment, the elapsed time for 50% of the exposed clams to re-burrow into the sediment (RT(50)) was also calculated in a 24-h re-burrowing response test. At the highest NP concentrations, it was not possible to estimate RT(50) values, as the percentage of clams showing complete burial was less than 50%. The present study demonstrates that NP causes oxidative stress by inhibiting superoxide dismutase activity and greatly affects the re-burrowing capability of T. philippinarum, even at environmentally realistic concentrations.     

Use of exposure time and life expectancy in models for toxicity to aquatic organisms

The exposure time is a variable which is usually not incorporated into models for toxicity. However, with persistent organic pollutants (POPs) exhibiting a nonspecific mode of action and lipophilic properties this variable can be modeled by the usage of the internal concentrations as a measure of the toxicity with fish. The bioconcentration process with fish is a relatively well understood and predictable process which allows the calculation of the internal lethal concentration. When the exposure time is relatively short the critical internal lethal concentrations are relatively constant for the group of POPs whereaas the LC50 measured in the ambient water is quite variable. When the exposure time is relatively long, results on the measurement of the critical internal concentration with fish over different exposure times has demonstrated that the internal lethal concentration falls with increasing exposure times in a consistent and predictable manner. This reduction in life expectancy can be described in a model which can be used to estimate the critical internal concentration for any exposure time. It also provides information useful in assessing the risk to fish and potentially other species due to the occurrence of residues of POPs in natural aquatic systems. It is suggested that these relationships can be extended to other groups of organisms and chemicals.     

Use of exposure time and life expectancy in models for toxicity to aquatic organisms

The exposure time is a variable which is usually not incorporated into models for toxicity. However, with persistent organic pollutants (POPs) exhibiting a nonspecific mode of action and lipophilic properties this variable can be modeled by the usage of the internal concentrations as a measure of the toxicity with fish. The bioconcentration process with fish is a relatively well understood and predictable process which allows the calculation of the internal lethal concentration. When the exposure time is relatively short the critical internal lethal concentrations are relatively constant for the group of POPs whereaas the LC50 measured in the ambient water is quite variable. When the exposure time is relatively long, results on the measurement of the critical internal concentration with fish over different exposure times has demonstrated that the internal lethal concentration falls with increasing exposure times in a consistent and predictable manner. This reduction in life expectancy can be described in a model which can be used to estimate the critical internal concentration for any exposure time. It also provides information useful in assessing the risk to fish and potentially other species due to the occurrence of residues of POPs in natural aquatic systems. It is suggested that these relationships can be extended to other groups of organisms and chemicals.     

Determination of both exposure time and denudation rate from an in situ-produced 10Be depth profile: A mathematical proof of uniqueness. Model sensitivity and applications to natural cases

Measurements of radioactive in situ-produced cosmogenic nuclide concentrations in surficial material exposed to cosmic rays allow either determining the long-term denudation rate assuming that the surface studied has reached steady-state (where production and losses by denudation and radioactive decay are in equilibrium) (infinite exposure time), or dating the initiation of exposure to cosmic rays, assuming that the denudation and post-depositional processes are negligible. Criteria for determining whether a surface is eroding or undergoing burial as well as quantitative information on denudation or burial rates may be obtained from cosmogenic nuclide depth profiles. With the refinement of the physical parameters involved in the production of in situ-produced cosmogenic nuclides, a unique well-constrained depth profile now permits determination of both the exposure time and the denudation rate affecting a surface. In this paper, we first mathematically demonstrate that the exponential decrease of the in situ-produced ¹⁰Be concentrations observed along a depth profile constrains a unique exposure time and denudation rate when considering both neutrons and muons. In the second part, an improved chi-square inversion model is described and tested in the third part with actual measured profiles.     

An assessment of marine pollution in Puget Sound

The Marine Ecosystems Analysis (MESA) Puget Sound Project was initiated to determine the existing levels, sources, fates and effects of toxic chemicals in Puget Sound, Washington. An approach was developed to assess the concentrations of a suite of contaminants and to measure a suite of biological parameters known to be responsive to contaminants. Metals, aromatic hydrocarbons and synthetic organics were found throughout the Sound. Highest concentrations were in bays near urban areas, particularly Elliott Bay near Seattle and Commencement Bay near Tacoma. A variety of biological disorders, including liver tumors in flatfish, and benthic community parameters were correlated with elevated contaminant levels. Because of the high contaminant concentrations there, and the high potential for exposure of humans to them, Commencement Bay has been identified by the US Environmental Protection Agency as a high priority toxic dump site in the United States, and remedial action is planned.     

Inhibition of coral photosynthesis by the antifouling herbicide Irgarol 1051

International regulation of organotin compounds for use in antifouling paints has led to the development and increased use of replacement compounds, notably the s-triazine herbicide Irgarol 1051. Little is known about the distribution of Irgarol 1051 in tropical waters. Nor has the potential impact of this triazine upon photosynthesis of endosymbiotic microalgae (zooxanthellae) in corals been assessed. In this study Irgarol 1051 was detected in marinas, harbours and coastal waters of the Florida Keys, Bermuda and St. Croix, with concentrations ranging between 3 and 294 ng 1(-1). 14C incubation experiments with isolated zooxanthellae from the common inshore coral Madracis mirabilis showed no incorporation of H14CO3- from the sea water medium after 4-8 h exposure to Irgarol 1051 concentrations as low as 63 ng 1(-1). Reduction in net photosynthesis of intact corals was found at concentrations of l00 ng 1(-1) with little or no photosynthesis at concentrations exceeding 1000 ng 1(-1) after 2-8 h exposure at all irradiances. The data suggest Irgarol 1051 to be both prevalent in tropical marine ecosystems and a potent inhibitor of coral photosynthesis at environmentally relevant concentrations.     

Some Haematological Effects of Crude Oil on Freshwater Catfish,Heteropneustes fossilis (BLOCH)

The exposure of catfish, Heteropneustes fossilis to different concentrations (200, 500, 700, 1000, 1500, 2000 ppm) of crude oil extract for a varying period (for up to 48 h in lethal extract and 360 h in sublethal one) resulted into a number of haematological changes. All the parameters taken here were found to have been increased except the haemoglobin level which fell down to 14.3 % in an acute lethal concentration (2000 ppm) after 48 h of exposure. The haematocrit value increased significantly (12 %) in 2000 ppm. The blood sugar level showed hyperglycemia in all the concentrations. The increase in ascorbic acid (28 %) was pronounced in higher concentrations. Changes occurring in different parameters seemed to be reversible as all the parameters returned to their normal levels after returning the fish to normal media, except the nuclear swelling which did not resume the normal functioning even after a prolonged treatment (1 month or more) in the recovery jar. The haematological effects shown by the crude oil resembled partly those kept in severe hypoxic conditions and partly to the fishes poisoned by heavy metals (Cu and Zn). On the basis of results obtained in this investigation, crude oil may be categorised as a complex toxic agent.     

The Impact of the Herbicide Diuron on Photosynthesis in Three Species of Tropical Seagrass

The impact and recovery from exposure to the herbicide diuron [DCMU; 3-(3′,4′-dichlorophenyl)-1,1-dimethylurea] was assessed for three tropical seagrasses, maintained in outdoor aquaria over a 10-day period. Photosynthetic stress was detected using chlorophyll a fluorescence, measured with a Diving-PAM (pulse amplitude modulated fluorometer). Exposure to 10 and 100 μg l⁻¹ diuron resulted in a decline in effective quantum yield (ΔF/F_m^′) within 2 h of herbicide exposure in Cymodocea serrulata, Halophila ovalis and Zostera capricorni. Effective quantum yield also declined over the first 24 h of exposure in H. ovalis at even lower diuron concentrations (0.1 and 1.0 μg l⁻¹). Effective quantum yield in H. ovalis and Z. capricorni was significantly depressed at all diuron concentrations (0.1–100 μg l⁻¹) after 5 days exposure, whereas effective quantum yield in C. serrulata was only significantly lower in plants exposed to highest diuron concentrations (10 and 100 μg l⁻¹). Effective quantum yield depression was present 5 days after plants exposed to 10 and 100 μg l⁻¹ diuron were returned to fresh seawater. These results indicate that exposure to herbicide concentrations present in nearshore Queensland sediments present a potential risk to seagrass functioning.     

Radon-222 Concentration and Aquifer Lithology in North Carolina

The presence of the radioactive gas radon (Rn-222) in many ground water supplies is a potentially significant source of public exposure to ionizing radiation. A wide range of radon concentrations has been measured in ground water in North Carolina, including some far in excess of national average concentrations. North Carolina is, however, geologically complex and ground water radon concentrations vary considerably among the state's aquifers. The highest average radon concentrations occur in areas underlain by granites (geometric mean 5910 pCi/l), and the lowest occur in the Atlantic Coastal Plain region (48 pCi/l). Average radon levels intermediate between these extremes are characteristic of the large areas of North Carolina underlain by gneisses, schists and metavolcanic rocks. Relative average radon concentrations in ground water from the rock types surveyed are consistent with relative abundances of uranium, the parent element of radon, in these rocks. Although other geologic and hydrologic factors also have an effect, aquifer lithology is a useful predictor of the concentration of radon in ground water. The occurrence of high radon concentrations in certain aquifer types; such as granites, shows that geologic factors should be considered in estimates of population exposure to radon, and that knowledge of aquifer geology can help to predict ground water radon concentrations in areas where field sampling has not been done.