Histochemical and biochemical markers in trout larvae exposed to river sediments

Rainbow trout larvae (Oncorhynchus mykiss) were exposed to bottom sediments collected from 4 sites of the River Po, located close to the springs (UP) of the main Italian river and downstream the confluence of 3 tributaries of its middle reach, the Rivers Ticino (TI), Adda (AD) and Lambro (LA), which drain areas characterized by increasing anthropogenic stress. After 7 days of exposure the trout larvae were examined for both biochemical and histochemical markers and results indicate the presence of a toxicological risk for the fish community of the AD- and particularly LA-downstream stretches. The reach of the River Po receiving the Ticino River was confirmed to be less hazardous in conjunction with the UP area.     

Assessment of the environmental significance of heavy metal pollution in surficial sediments of the River Po

The magnitude and ecological relevance of metal pollution of the middle Po river deriving from the River Lambro tributary was investigated by applying different (complementary) sediment quality assessment approaches: (1) comparisons of concentrations with regional reference data, and (2) comparisons with consensus-based sediment quality guidelines (SQGs), as well as by investigations of the partitioning patterns of target heavy metals (Cd, Cu, Ni, Pb, Zn). Total metal concentrations in the surficial sediments revealed significant pollution inputs on the whole river stretch investigated, with a distinct peak at the inlet of the River Lambro. Based on the geoaccumulation index of target heavy metals, the middle reach of River Po has to be considered as moderately polluted with Cd (1相似文献   

Androgenic and antiandrogenic activities in water and sediment samples from the river Lambro, Italy, detected by yeast androgen screen and chemical analyses

The river Lambro is the most polluted tributary of the river Po in North Italy and was chosen as a representative water course discharging industrialized areas. Water and sediment samples of the river Lambro were investigated regarding the presence of endocrine disrupting compounds. A combined procedure was used consisting of solid-phase extraction and HPLC based fractionation of samples, of screening for (anti)androgenic activity using the yeast androgen screen (YAS) and of chemical analysis using HPLC-MS/MS and GC-MS. Androgenic and antiandrogenic activities were found in specific fractions of the water and sediment while the total extracts showed antiandrogenic activity only. The chemical analysis of the fractions and total extracts with antiandrogenic activities revealed the presence of compounds with suspected antiandrogenic potency such as bisphenol A, iprodione, nonylphenol, p,p'-DDE and tert-octylphenol but other unknown compounds contributed mainly to the observed antiandrogenic activities. The antiandrogenic load of the river Lambro ranged between 1.34 and 17.1 microM flutamide-equivalents and may pose a risk to aquatic environments. Future screenings for EDC in the environment that have the potential to interfere with reproduction of aquatic organisms should be extended to different modes of actions including (anti)androgenic ones.     

Ah Receptor Agonists in UV-exposed Toluene Solutions of Decabromodiphenyl Ether (decaBDE) and in Soils Contaminated with Polybrominated Diphenyl Ethers (PBDEs) (9 pp)

Goal, Scope and Background The use of polybrominated diphenyl ethers (PBDEs) as flame retardants increases the risk for emissions of other brominated compounds, such as polybrominated dibenzodioxins (PBDDs) and dibenzofurans (PBDFs). The large homology in structure of PBDD/Fs and mechanism of toxic action, i.e. the capacity to activate the Ah receptor (AhR) pathway, compared to their well-studied chlorinated analogues, justifies a raised concern to study the environmental levels and fate of these compounds. Decabromodiphenyl ether (decaBDE) is the most widely used PBDE today. Studies on photolytic debromination of decaBDE in organic solvents have shown debromination of decaBDE, as well as formation of PBDFs. However, little is known about the transformation mechanisms and there are only scarce data on photoproducts and PBDE transformation in environmentally relevant matrices. In this study, mechanism-specific dioxin bioassays were used to study photolytic formation of AhR agonists in toluene solutions of decaBDE. In addition, the influence of irradiation time and UV-light wavelength on the formation was studied. PBDE congener patterns and presence of PBDD/Fs were analysed. Further, AhR agonists were analysed in agricultural soils contaminated with PBDEs. Soils were also exposed to UV-light to study changes in AhR agonist levels. Methods Toluene solutions of decaBDE were irradiated using three different spectra of UV-light, simulating UV-A (320-400 nm), UV-AB (280-400 nm), and UV-ABC (250-400 nm). Additionally, decaBDE solutions were exposed to narrow wavelength intervals (10 nm bandwidth) with the central wavelengths 280, 290, 300, 310, 320, 330, 340, 350, 360 nm. AhR agonists in decaBDE solutions were analysed with two different bioassays, the chick embryo liver-cell assay for dioxins (Celcad) and the dioxin responsive, chemically activated luciferase expression assay (DR-Calux). Also, the decaBDE solutions were analysed with LRGC-LRMS to obtain PBDE congener patterns for breakdown of decaBDE, and with HRGC-HRMS, for presence of PBDD/Fs. Four soils were exposed to UV-AB light, under both dry and moist conditions. Levels of AhR agonists in soil extract fractions, before and after UV-exposure, were analysed with the DR-Calux. Results and Discussion Significant levels of photoproducts able to activate the AhR pathway, up to 31 ng bio-TEQ/ml, were formed in UV-exposed decaBDE solutions. The transformation yield of decaBDE into AhR agonists was estimated to be at the 0.1%-level, on a molar basis. The net formation was highly dependent on wavelength, with the sample irradiated at 330 nm showing the highest level of dioxin-like activity. No activity was detected in controls. PBDE analysis confirmed decaBDE degradation and a clear time-dependent pattern for debromination of PBDE congeners. AhR agonist effect in the recalcitrant fractions of the soils corresponded to the levels of chemically derived TEQs, based only on chlorinated dioxin-like compounds in an earlier study. It was concluded that no significant levels of other AhR agonists, e.g. PBDFs, were accumulated in the soil. UV-light caused changes in AhR-mediated activity in the more polar and less persistent fractions of the soils, but it is not known which compounds are responsible for this. Recommendations and Perspective . The laboratory experiments in this study show that high levels of AhR agonists can be formed as photoproducts of decaBDE and it is important to elucidate if and under which conditions this might occur in nature. However, soil analysis indicates that photoproducts of PBDE do not contribute to the accumulated levels of persistent dioxin-like compounds in agricultural soil. Still, more data is needed to fully estimate the environmental importance of PBDE photolysis and occurrence of its photoproducts in other environmental compartments. Analysis with dioxin bioassays enabled us to gather information about photoproducts formed from decaBDE even though the exact identities of these compounds were not known. Conclusion Bioassays are valuable for studying environmental transformation processes like this, where chemical analysis and subsequent toxicological evaluation requires available standard compounds and information on toxicological potency. The use of bioassays allows a rapid evaluation of toxicological relevance.     

Bioavailability of polychlorinated dibenzo-p-dioxins and dibenzofurans from contaminated Wisconsin River sediment to carp

The bioavailability of 2,3,7,8-TCDD from sediment to freshwater fish was studied in laboratory exposures. Carp (10g) exposed to Wisconsin River sediment (39 pg/g) for 55 days accumulated 7.5 pg/g. Maintaining exposured fish in clean water for an additional 205 days resulted in depuration of 32–34% TCDD. These values compare to field data where sediments ranged from 30–200 pg/g and 1.5 kg carp (70 pg/g) depurated 55% in 325 days. Analysis of sediment, laboratory exposed fish, and Wisconsin River fish for other PCDDs and PCDFs showed residues of 2,3,7 and 8 substituted congeners selectively enriched.     

Analysis of perfluorooctanoate (PFOA) and other perfluorinated compounds (PFCs) in the River Po watershed in N-Italy

C7-C11 perfluorinated carboxylates (PFACs) and perfluorooctansulfonate (PFOS) were analysed in selected stretches of the River Po and its major tributaries. Analyses were performed by solid-phase extraction (SPE) with Oasis HLB cartridges and methanol elution followed by LC-MS-MS detection using 13C-labelled internal standards. High concentration levels ( approximately 1.3 microg l(-1)) of perfluorooctanoate (PFOA) were detected in the Tánaro River close to the city Alessandria. After this tributary, levels between 60 and 337 ng l(-1) were measured in the Po River on several occasions. The PFOA concentration close to the river mouth in Ferrara was between 60 and 174 ng l(-1). Using the river discharge flow data in m3 s(-1) at this point (average approximately 920 m3 s(-1) for the year 2006), a mass load of approximately 0.3 kg PFOA per hour or approximately 2.6 tons per year discharged in the Adriatic Sea has been calculated. PFOS concentration levels in the Po River at Ferrara were approximately 10 ng l(-1).     

Acute toxicity of copper,lead, cadmium,and zinc to early life stages of white sturgeon (Acipenser transmontanus) in laboratory and Columbia River water

Populations of white sturgeon (Acipenser transmontanus) are in decline in North America. This is attributed, primarily, to poor recruitment, and white sturgeon are listed as threatened or endangered in several parts of British Columbia, Canada, and the United States. In the Columbia River, effects of metals have been hypothesized as possible contributing factors. Previous work has demonstrated that early life stage white sturgeon are particularly sensitive to certain metals, and concerns over the level of protectiveness of water quality standards are justified. Here we report results from acute (96-h) toxicity tests for copper (Cu), cadmium (Cd), zinc (Zn), and lead (Pb) from parallel studies that were conducted in laboratory water and in the field with Columbia River water. Water effect ratios (WERs) and sensitivity parameters (i.e., median lethal accumulations, or LA50s) were calculated to assess relative bioavailability of these metals in Columbia River water compared to laboratory water, and to elucidate possible differences in sensitivity of early life stage white sturgeon to the same concentrations of metals when tested in the different water sources. For Cu and Pb, white sturgeon toxicity tests were initiated at two life stages, 8 and 40 days post-hatch (dph), and median lethal concentrations (LC50s) ranged between 9–25 μg Cu/L and 177–1,556 μg Pb/L. LC50s for 8 dph white sturgeon exposed to Cd in laboratory water and river water were 14.5 and 72 μg/L, respectively. Exposure of 8 dph white sturgeon to Zn in laboratory and river water resulted in LC50s of 150 and 625 μg/L, respectively. Threshold concentrations were consistently less in laboratory water compared with river water, and as a result, WERs were greater than 1 in all cases. In addition, LA50s were consistently greater in river water exposures compared with laboratory exposures in all paired tests. These results, in combination with results from the biotic ligand model, suggest that the observed differences in toxicity between river water exposures and laboratory water exposures were not entirely due to differences in water quality and metal bioavailability but rather in combination with differences in fish sensitivity. It is hypothesized that differences in concentrations of calcium in the different water sources might have resulted in differences in acquired sensitivity of sturgeon to metals. Canadian water quality guidelines, US national criteria for the protection of aquatic life, and water quality criteria for the state of Washington were less than LC50 values for all metals and life stages tested in laboratory and Columbia River water. With the exception, however, that 40 dph white sturgeon exposed to Cu in laboratory water resulted in threshold values that bordered US national criteria and criteria for the state of Washington.     

Polychlorinated dibenzo-p-dioxins and dibenzofurans in River Po sediments

The River Po is the main Italian river draining one of the most populated and industrialised regions in Italy. As part of a monitoring project to assess environmental quality in the River Po, we measured the concentrations of polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) in bottom sediments collected along the whole course of the river, from the spring to the delta, downstream from the confluence of its main tributaries. The aim was to investigate the level of contamination in the main Italian river and the contribution of contaminant loads from the subbasins. Composite sediment samples were collected in summer and winter surveys in low-flow water conditions and analysed by HRGC-HRMS for PCDD and PCDF homologue groups and for the 2,3,7,8-substituted congeners. The spatial trend observed in the Po River for PCDD and PCDF concentrations varied depending on the load of contaminants received from the tributaries and the processes of sedimentation. The sum of PCDD and PCDF concentrations, and the toxic equivalent content, ranged from 121 to 814 and from 1.3 to 13 ng/kg dry weight sediment, respectively. These levels of contamination seem lower than in the sediments of rivers draining highly industrialised areas. The PCDD and PCDF homologue profiles in all the samples were very similar, suggesting a common source of this contamination in the River Po. Principal component analysis suggested that widespread sources, such as urban runoff and domestic wastewaters, are probably the main cause of these levels of PCDDs and PCDFs.     

Acute and Chronic Toxicity of Chlorine Dioxide (ClO2) and Chlorite (ClO2ˉ) to Rainbow Trout (Oncorhynchus mykiss) (4 pp)

Goal, Scope and Background Chlorite (ClO2ˉ) is a primary decomposition product when chlorine dioxide (ClO2) is added during water treatment; therefore the toxic effects of both compounds on aquatic organisms are possible. Limited data are available concerning their toxicity to fish. The aim of this study was to investigate sensitivity of rainbow trout to acute and chronic toxicity of chlorine dioxide and chlorite, and to estimate the Maximum-Acceptable-Toxicant-Concentration (MATC) of those compounds in fish. Methods The acute and chronic toxicity of chlorine dioxide and chlorite to larval and adult rainbow trout was investigated in 96-hour to 20-day laboratory exposures evaluating the wide range spectrum of biological indices under semi-static conditions. Results and Discussion Median lethal concentration (96-hour LC50) values derived from the tests were: 2.2 mg/l for larvae; 8.3 mg/l for adult fish and 20-day LC50 for larvae was 1.6 mg/l of chlorine dioxide, respectively. Chlorite was found to be from 48 to 18 times less acutely toxic to larvae and adult fish, correspondingly. Both chemical compounds induced similar toxic effects in rainbow trout larvae during chronic tests (they affected cardio-respiratory and growth parameters), but chlorine dioxide had a higher toxic potency than chlorite. A significant decrease in the heart rate and respiration frequency of larvae was established. However, within an increase in exposure duration recovery of cardio-respiratory responses was seen to have occurred in larvae exposed to chlorite. Meanwhile, in larvae exposed to chlorine dioxide, a significant decrease in cardio-respiratory responses remained during all 20-day chronic bioassays. Chlorine dioxide also more strongly affected growth parameters of rainbow trout larvae at much lower test concentrations. Decreased rate of yolk-sack resorption occurred only in the tests with chlorine dioxide. Conclusions Maximum-Acceptable-Toxicant-Concentration (MATC) of 0.21 mg/l for chlorine dioxide and of 3.3 mg/l for chlorite to fish was derived from chronic tests based on the most sensitive parameter of rainbow trout larvae (growth rate). According to substance toxicity classification accepted for Lithuanian inland waters, chlorine dioxide and chlorite can be referred to substances of \moderate\ toxicity to fish. Recommendations and Outlook Due to its very reactive nature, chlorine dioxide is rapidly (in a few hours) reduced to chlorite, which is persistent also as a biocide but 16 times less toxic to fish, according to MATC. Therefore, it is much more likely that fish will be exposed to chlorite than to chlorine dioxide in natural waters. Presently accepted, the Maximum-Permitted-Concentration of total residual chlorine (TRC) in waste-water discharging into receiving waters is 0.6 mg/l. If this requirement will not be exceeded, it is unlikely that fish would be exposed to lethal or even to sublethal concentrations of chlorine dioxide or chlorite. Furthermore, chlorine dioxide does not generate toxic nitrogenous (chloramines) or carcinogenic organic residuals (trihalomethanes). All these properties make chlorine dioxide a more promising biocide than chlorine.     

Characteristics of organic phosphorus fractions in different trophic sediments of lakes from the middle and lower reaches of Yangtze River region and Southwestern Plateau, China

In this study, the characteristics of organic phosphorus (Po) fractions in sediments of six lakes from the middle and lower reaches of Yangtze River region and Southwestern China Plateau, China were investigated using a soil Po fractionation scheme, and the relationships between Po, inorganic phosphorus (Pi) and pollution status were also discussed. The results show that the rank order of Po fractions was: residual Po>HCl-Po>fulvic acid-P>humic acid-P>NaHCO3-Po, with their average relative proportion 8.7:4.6:3.2:2.1:1.0. Po fractions, especially nonlabile Po, were significantly correlated with organic matter, Po and NaOH-Pi. Different distribution patterns of P fractions were observed in those two different regions. Po fractions in the heavily polluted sediments were higher than those in moderately and no polluted sediments, it is suggested that Po should be paid more attention in the lake eutrophication investigation.     

Applying adult emergence as an endpoint in a post-exposure laboratory test using two midge species (Diptera: Chironomidae)

Several approaches have been used to evaluate biological impairment in aquatic ecosystems which can be categorized as either laboratory or field. In the recent years, the laboratory toxicity test approach has been extended to field exposures where ambient factors are allowed to influence the test response. Field exposures of laboratory test organisms require method modifications. In this paper, a novel in situ method is described which measures growth, survival and emergence of sediment inhabiting insects (Diptera: Chironomidae) that are used in standardized laboratory toxicity testing. Two standard chironomid species (Chironomus riparius and Chironomus tentans) were used to test the suitability of the approach and to compare the performance of the species. The larvae were transferred to the laboratory for emergence after 7 days in situ exposure which was compared to laboratory responses. Growth, survival and emergence were significantly lower in the in situ pre-exposure than in the laboratory. Also, emergence success was significantly lower in one reference sediment (LMR) than in the other test sediments in both in situ and the laboratory treatment. These lower response levels likely resulted from sediment characteristics and artifacts related to the exposure in the in situ chamber. Feeding and water quality within the exposure chamber appear to be factors that may differ markedly from the laboratory exposure and may affect organism responses. C. riparius developed (growth, emergence time) faster than C. tentans in all treatments, otherwise the species responded similarly. C. riparius may be a better alternative for the chronic in situ exposures because of shorter exposure times and reduced feeding requirements.     

Effects of pH and feeding regime on methylmercury accumulation within aquatic microcosms

The effect of pH (pH 5, 6 or 7) on accumulation of radiolabelled methylmercury was examined using a laboratory microcosm system. Accumulation of labelled mercury at three trophic levels, primary consumers (Daphnia magna), secondary consumers (rainbow trout, Salmo gairdneri) and tertiary consumers (walleye, Stizostedion vitreum) was not significantly affected by pH. Our results are in direct contrast with field observations of elevated methylmercury concentrations in fish from low pH water and indicate that the elevated mercury residues observed in the field result from factors other than the direct effects of pH on accumulation of methylmercury by aquatic organisms. Both water and diet were important as sources of the labelled mercury which was accumulated by walleye. Walleye which were fed rainbow trout, that had accumulated labelled mercury from within the experimental microcosms, accumulated almost twice as much labelled mercury as walleye fed non-labelled prey, or walleye which were not fed. Walleye fed non-labelled rainbow trout accumulated slightly more labelled mercury than walleye which were not fed, presumably as a result of the higher metabolic rate of the fish which were fed.     

A study of polonium-210 distribution aspects in the riverine ecosystem of Kaveri, Tiruchirappalli, India

Distribution of a natural radionuclide (210Po) in water, sediment and biota was quantified along a 95 km reach of the Kaveri River, India. It is observed that the soft tissues of animals concentrated a higher level of 210Po than the hard parts like shell and bone. The ranges of 210Po activity in biological samples were 2.32-10.78 Bq kg(-1) wet (aquatic weeds), 18.94-28.55 Bq kg(-1) wet (plankton), 32.51 to 46.17 Bq kg(-1) wet (snail), 57.42 to 105.78 Bq kg(-1) wet (bivalve), 12.13 to 19.10 Bq kg(-1) wet (prawn) and 1.86 to 4.17 Bq kg(-1) wet (fish). The concentration factors (CFs) for the aquatic organisms ranged from 10(3) to 10(4), indicating a high affinity of 210Po for organic moieties. The bivalve mollusc, Lamellidens marginalis, accumulated higher concentrations of 210Po, suggesting that it could serve as a biomonitor of 210Po radionuclide in riverine systems. Further, the levels of 210Po in water, sediment and biological samples from impoundments were higher than those from stations located in the open river. This may be largely due to aerial inputs of the radionuclide, accumulation of radionucliderich silt and organic matter, and increased biological production in the impounded water body.     

Sediment toxicity and stormwater runoff in a contaminated receiving system: consideration of different bioassays in the laboratory and field.

Several field and laboratory assays were employed below an urban storm sewer outfall to define the relationship between stormwater runoff and contaminant effects. Specifically, two bioassays that measure feeding rate as a toxicological endpoint were employed in the field and in the laboratory, along with bioassays measuring survival and growth of test organisms. In 7 to 10 d in situ exposures, amphipod leaf disc processing, growth and survival were monitored. Different exposure scenarios were investigated by varying the mesh size (74 microns or 250 microns mesh) and method of deployment (water column, sediment surface, or containing sediment) of in situ exposure chambers. Hyalella azteca, Daphnia magna, and Pimephales promelas survival were monitored in 48 h in situ exposures. Feeding inhibition was investigated via enzyme inhibition of H. azteca and D. magna and via leaf disc processing measurements of the detritivore H. azteca. Additionally, we investigated the extent of phototoxicity at this site via field exposures in sun and shade and laboratory exposures with and without UV light. The measurement of detritivore leaf disc processing, and thus its usefulness as an endpoint, was hindered by individual variability in the amount of leaf consumed and by leaf weight gain during the summer field exposures. For D. magna, enzyme inhibition measured in a laboratory exposure did not reveal the toxicity observed in field exposures. For H. azteca, enzyme inhibition measured in the laboratory indicated toxicity similar to that observed in short term chronic in situ exposures. Enzyme inhibition also did not detect differences in toxicity due to variations in flow conditions. There were no statistically significant effects of any exposure on P. promelas survival or H. azteca growth, and there were no statistically significant effects due to mesh size or sun exposure. Survival of H. azteca was the most sensitive and the least variable endpoint. Effects on survival were noted in the same treatments over short-term chronic exposures in the laboratory and in situ. Significant differences in survival were noted due to the method of deployment under low flow conditions. In situ chambers containing sediment resulted in greater mortality in the 10 d low flow in situ experiments. Under high flow conditions, significant reductions in survival and leaf disc processing were noted under all methods of deployment at the two impacted sites over a 7 d exposure. Also under high flow conditions, significantly greater mortality of H. azteca was reported at the downstream field site when sediment was included in the chamber at deployment. These results suggest that significant toxicity at this site is due to accumulation of contaminants in the sediment and the mobilization of these contaminants during a storm event. In situ exposures detected toxicity not observed in laboratory exposures. These results suggest that a combination of laboratory and field bioassays is most useful in defining field effects.     

Environmental contaminants and biochemical response in eel exposed to Po river water

Groups of eels (Anguilla anguilla) were exposed to Po river water under control conditions at "la Casella" Fluvial Hydrobiology Station for 30-day periods between November and May. At the end of the exposure period, fish were sacrificed and cytochrome P4501A was evaluated in liver microsomes using both catalytic (EROD) and semiquantitative immunodetection (ELISA) assays. At the same time, water samples were taken for chemical analyses of PCBs, PAHs and pesticides. Eel was chosen as bio-indicator on the bases of experiments on cytochrome P450 expression and activity under basal or induced conditions and in consideration of the ecological and economic relevance and ease of handling. Low levels of cytochrome P450 during Winter were followed by a step increase during Spring, associated with a substantial concentration increase of agrochemicals. A good correlation was found between measurements of cytochrome P4501A by EROD and ELISA in dose-effect experiments, however, ELISA showed a higher sensitivity. Immunochemical techniques may be used in addition to enzyme activity measurements both to detect lower levels of cytochrome P450 induction (where they proved more sensitive) and as quality control. These results suggest that measurement of cytochrome P4501A under controlled conditions, using eel as bio-indicator, can be a useful tool in monitoring Po river ecosystems.     

Effects of mercury exposure on ionic regulation in the crayfish Orconectes propinquus

Hemolymph sodium, potassium and calcium concentrations were determined in crayfish (Orconectes propinquus) exposed to (203)HgCl(2) mixed with food to a concentration of 1 microg Hg g(-1). Dummy-fed animals were exposed to Hg-dosed food wrapped in dialysis tubing to control for mercury reaching the animals via leaching from food to water. Hemolymph analyses were made following 14-day Hg exposures and again after a further 21-day 'depuration' period during which all animals were fed Hg-free food. After 14 days, the mercury reached a concentration of 0.175 microg g(-1) in the hepatopancreas and approximately half this level in the gills of Hg-fed animals. No depuration occurred from the hepatopancreas although the gills lost approximately two-thirds of their labelled mercury during the depuration period. Hemolymph sodium concentrations in Hg-exposed crayfish, both fed and dummy-fed, after 14 days were significantly lower than in Hg-free controls and remained low following the 21-day depuration period. Hemolymph calcium concentrations were lower in Hg-fed animals than in dummy-fed and control animals after 14 days although calcium levels rose in all treatments after 35 days. This may have been due to the incidence of pre-molt animals in all experimental groups, although the relationship between this and mercury exposure was not established unequivocally. Hemolymph potassium levels showed no differences between treatments.     

Toxic Effects of Crude Oil Combined with Oil Cleaner Simple Green on Yolk-sac Larvae and Adult Rainbow Trout Oncorhynchus mykiss (4 pp)

Background, Goal and Scope Cleaner CRYSTAL Simple Green (SG) was used for the cleanup of the oil spill in the Baltic Sea near Lithuania in 2001. No scientific data are available on the effects and consequences of its application for local aquatic life. The aim of this study was to determine and compare sublethal effects of a) solution SG; b) crude oil alone; c) SG in combination with oil on rainbow trout Oncorhynchus mykiss at different stages of its development in laboratory conditions.Methods Laboratory studies were performed on adult rainbow trout (4-day duration) and on yolk-sac larvae (25-day duration) evaluating their biological parameters. Concentrations of water-soluble and thin-dispersed fractions of petroleum hydrocarbons were measured using gas chromatography.Results and Discussion SG solution (0.5 mg/l) did not affect the survival of larvae and adult fish, and no significant changes were determined in respiratory parameters of the exposed larvae and adult fish. The most expressed alterations were found in morphological parameters (a decrease in the average body mass) of larvae and in haematological indices (a decrease in the leukocyte count) of adult fish at the end of the tests. Crude oil (1610 mg/l) did not affect the survival of adult fish during the 4-day exposure. An increase in larvae mortality rate (~ 36%) was recorded at the end of the tests. A significant decrease in the average body mass and heart rate of larvae as well as in gill ventilation frequency of larvae and adult fish were determined. SG combined with oil induced an increase in larval mortality  46% of individuals died at the end of the tests. No mortality was recorded in adult fish. The average body mass and heart rate of larvae were significantly decreased. Marked changes were also found in respiratory parameters (gill ventilation frequency of larvae and adult fish significantly decreased, while “coughing” rate increased). A 1-day, 2-day exposure of fish to SG combined with oil induced a significant decrease in the leukocyte count of adult fish, which was also determined at the end of the tests. The augmentation of adverse impact could be explained by the data obtained from our studies. When SG was added into dilution water with crude oil the concentration of petroleum hydrocarbons in the mixture increased 3 ~ 4.5 times after 24 h and 96 h, respectively. Conclusions The comparative study of the effects of crude oil alone, SG and SG combined with oil showed that their toxic effects on fish differed. Oil combined with SG was found to be more toxic to fish (larvae and adults) than SG alone and oil alone. Fish at early stages of development (yolk-sac larvae) were more sensitive to the effects of the compounds studied than adults.Recommendations and Outlook To diminish the negative impact of oil spill cleanup using chemicals on aquatic ecosystems, it is recommended to carry out more comprehensive studies of their effects and after-effects in laboratory conditions using a wide scale of local aquatic organisms. The selected species of the most sensitive aquatic organisms should include those which are unable to escape the impact of combined action of oil and cleaners. Special attention should be directed to the research of effects of these pollutants on studied organisms at their most sensitive stages of life (reproduction, hatching, early stages of development), as after-effects of exposure to pollutants may be observed in future generations.