Changes in gill morphology of Atlantic salmon (Salmo salar) smolts due to addition of acid and aluminum to stream water

One-year-old Atlantic salmon smolts were held in three artificial channels adjacent to a softwater (mean sp. cond. 30 microS cm(-1), circumneutral stream. Water in one channel was untreated (mean pH 6.25); the others received additions of acid (to mean pH 5.6), or acid plus aluminum (to mean pH 5.5; mean exchangeable Al 158 microg litre(-1)). Gills were sampled after 16 and 23 days of exposure for morphometric examination. On primary lamellae, chloride cells were more numerous in both experimental treatments than in controls. In contrast, numbers of chloride cells on secondary lamellae were elevated only in fish exposed to acid without added Al. Chloride cell size and shape also varied with time and treatment. Fewer gill mucous cells were found in fish exposed to acid plus Al than in controls. Chloride cell proliferation and structural changes may represent an attempt to compensate for increased ionic effluxes with low pH stress by increasing uptake. However, if Al concentrations are high, chloride cells do not proliferate along the secondary lamellae, or proliferating cells are damaged and lost. This may limit the potential to increase ionic uptake.     

Large variation in lipid content, SigmaPCB and delta13C within individual Atlantic salmon (Salmo salar)

Many studies that investigate pollutant levels, or use stable isotope ratios to define trophic level or animal origin, use different standard ways of sampling (dorsal, whole filet or whole body samples). This study shows that lipid content, SigmaPCB and delta(13)C display large differences within muscle samples taken from a single Atlantic salmon. Lipid- and PCB-content was lowest in tail muscles, intermediate in anterior-dorsal muscles and highest in the stomach (abdominal) muscle area. Stable isotopes of carbon (delta(13)C) showed a lipid accumulation in the stomach muscle area and a depletion in tail muscles. We conclude that it is important to choose an appropriate sample location within an animal based on what processes are to be studied. Care should be taken when attributing persistent pollutant levels or stable isotope data to specific environmental processes before controlling for within-animal variation in these variables.     

Avoidance of toxic mixing zones by Atlantic salmon (Salmo salar L.) and brown trout (Salmo trutta L.) in the limed River Audna, southern Norway

Chlorophyll a (chl a) fluorescence was used to determine the effects of treatments with gaseous HF or aqueous solutions of NaF on the photosynthetic apparatus of spinach prior to the appearance of visible injury. Placing the petioles in 2 mM NaF for 3 h resulted in the accumulation of 240 ppm F in leaf blades. The second oldest leaves of spinach plants accumulated similar concentrations (270 ppm F) when the plants were exposed to gaseous HF at 5 microg F m(-3) for 6 days. These NaF and HF treatments did not result in visible injury nor did they affect Fo, Fm or Fv/Fm. However, during the slow (>2 s) induction kinetics, fluorescence quenching in fluoride-treated leaves increased during the P to S phase and the M peak was no longer resolved. This was due, in part, to increased photochemical quenching. The results are consistent with a reduced ability to develop or maintain a trans-thylakoid proton gradient in chloroplasts containing elevated concentrations of F.     

Carry-over of dietary organochlorine pesticides, PCDD/Fs, PCBs, and brominated flame retardants to Atlantic salmon (Salmo salar L.) fillets

Information on carry-over of contaminants from feed to animal food products is essential for appropriate human risk assessment of feed contaminants. The carry-over of potentially hazardous persistent organic pollutants (POPs) from feed to fillet was assessed in consumption sized Atlantic salmon (Salmo salar). Relative carry-over (defined as the fraction of a certain dietary POP retained in the fillet) was assessed in a controlled feeding trial, which provided fillet retention of dietary organochlorine pesticides (OCPs), dioxins (PCDD/Fs), polychlorinated biphenyls (PCBs), and brominated flame retardants (BFRs). Highest retention was found for OCPs, BFRs and PCBs (31-58%), and the lowest retentions were observed for PCDD/Fs congeners (10-34%). National monitoring data on commercial fish feed and farmed Atlantic salmon on the Norwegian market were used to provide commercially relevant feed-to-fillet transfer factors (calculated as fillet POP level divided by feed POP level), which ranged from 0.4 to 0.5, which is a factor 5-10 times higher than reported for terrestrial meat products. For the OCP with one of the highest relative carry-over, toxaphene, uptake and elimination kinetics were established. Model simulations that are based on the uptake and elimination kinetics gave predicted levels that were in agreement with the measured values. Application of the model to the current EU upper limit for toxaphene in feed (50 μg kg⁻¹) gave maximum fillet levels of 22 μg kg⁻¹, which exceeds the estimated permissible level (21 μg kg⁻¹) for toxaphene in fish food samples in Norway.     

Tissue bioaccumulation patterns, xenobiotic biotransformation and steroid hormone levels in Atlantic salmon (Salmo salar) fed a diet containing perfluoroactane sulfonic or perfluorooctane carboxylic acids

In the present study, groups of juvenile Atlantic salmon (Salmo salar) were fed gelatine capsules containing fish-food spiked with PFOA or PFOS (0.2 mg kg⁻¹ fish) and solvent (methanol). The capsules were given at days 0, 3 and 6. Blood, liver and whole kidney samples were collected prior to exposure (no solvent control), and at days 2, 5, 8 and 14 after exposure (Note: that day 14 after exposure is equal to 7 d recovery period). We report on the differences in the tissue bioaccumulation patterns of PFOS and PFOA, in addition to tissue and compound differences in modulation pattern of biotransformation enzyme genes. We observed that the level of PFOS and PFOA increased in the blood, liver and kidney during the exposure period. Different PFOS and PFOA bioaccumulation patterns were observed in the kidney and liver during exposure- and after the recovery periods. Particularly, after the recovery period, PFOA levels in the kidney and liver tissues were almost at the control level. On the contrary, PFOS maintained an increase with tissue-specific differences, showing a higher bioaccumulation potential (also in the blood), compared with PFOA. While PFOS and PFOA produced an apparent time-dependent increase in kidney CYP3A, CYP1A1 and GST expression, similar effects were only temporary in the liver, significantly increasing at sampling day 2. PFOA and PFOS exposure resulted in significant decreases in plasma estrone, testosterone and cortisol levels at sampling day 2, and their effects differed with 17α-methyltestostrerone showing significant decrease by PFOA (also for cholesterol) and increase by PFOS. PFOA significantly increased estrone and testosterone, and no effects were observed for cortisol, 17α-methyltestosterone and cholesterol at sampling day 5. Overall, the changes in plasma steroid hormone levels parallel changes in CYP3A mRNA levels. Given that there are no known studies that have demonstrated such tissue differences in bioaccumulation patterns with associated differences in toxicological responses in any fish species or lower vertebrate, the present findings provide some potential insights and basis for a better understanding of the possible mechanisms of PFCs toxicity that need to be studied in more detail.     

Reproduction, larval growth, and reproductive development in African clawed frogs (Xenopus laevis) exposed to atrazine

Reproductive success and development of F2 offspring from F1 adult African clawed frogs (Xenopus laevis) exposed to atrazine throughout larval development and as sexually mature adults was examined. Larval X. laevis were exposed to one of four nominal concentrations of atrazine (0, 1, 10, 25 microg atrazine/l) beginning 96 hr after fertilization and continuing through two years post-metamorphosis. Clutch size and survival of offspring were used as measurement endpoints to gauge reproductive success of the F1 frogs. Larval survivorship and time to metamorphosis were used to gauge developmental success of the F2 offspring from atrazine-exposed frogs. Testes in F1 and F2 frogs were examined for incidence of anomalies, such as testicular ovarian follicles, and sex ratios in F2 offspring were investigated to determine if exposure to atrazine caused trans-generational effects (effects on F2 individuals due to exposure of F1 individuals). There were no effects of any of the studied concentrations of atrazine on clutch size of F1 frogs. There were also no effects on hatching success or time to metamorphosis. Sex ratios did not differ between F2 offspring among treatments. There was no evidence to suggest a transgenerational effect of atrazine on spawning success or reproductive development of X. laevis. This is consistent with the presence of robust populations of X. laevis in areas where they are exposed to atrazine that has been used for several decades for weed control in production of corn. Our observations also are consistent with the results of most other studies of frogs where no effects were found to be associated with exposure to atrazine. Our data do not support the hypothesis that atrazine significantly affects reproductive fitness and development of frogs.     

Mortality response and LC50 values for juvenile and adult crayfish, Procambarus clarkii exposed to Thiodan (insecticide), Treflan, MSMA, Oust (herbicides) and Cutrine-Plus (algicide)

Toxicities of three herbicides (Treflan, MSMA and Oust), an algicide (Cutrine-plus) and an insecticide (Thiodan) to juvenile (3.0-3.4 cm) and adult (9.0-10.0 cm) crayfish, Procambarus clarkii, were determined by 96 h static bioassays. Freshly prepared 0.01% and 1.0% aqueous stock solutions of these pesticides were diluted to desired concentrations. Mortalities were recorded each day up to 96 h. Aged tap-water was used for preparing all test solutions. Per cent mortalities were analyzed for linear regression and LC(50) values were computed by probit analysis. LC(50) values for juvenile P.clarkii in the descending order of toxicity were: 24 ppb Thiodan, 13 ppm Treflan, 101 ppm MSMA, 461 ppm Cutrine-plus, 12,174 ppm Oust; and for adults these values were: 423 ppb Thiodan, 26 ppm Treflan, 1019 ppm MSMA and 2945 ppm Cutrine-plus. No LC(50) values for adult crayfish could be computed for Oust herbicide which caused no mortalities up to 60,000 ppm concentration. Comparing the overall toxicities of these pesticides, Thiodan was the most toxic to all crayfish; followed by Treflan, MSMA, Cutrine-plus and Oust. Oust was more than 1/2 million times less toxic than Thiodan to juvenile crayfish. Published LC(50) values indicate that freshwater crayfish are more tolerant than marine decapods, Crangon septemspinosa and Mysidopsis bahia, to Thiodan insecticide and Treflan herbicide.     

Metallothionein induction, antioxidative responses, glycogen and growth changes in Tubifex tubifex (Oligochaete) exposed to the fungicide, fenhexamid

Laboratory studies were conducted to determine the effects of different concentrations of fenhexamid (0.1, 1, and 10 mg L(-1)) on growth, oxidative stress, protein, glycogen, and metallothionein (MT) contents in Tubifex tubifex after an exposure of 2, 4, and 7 days. In addition, residues of the fungicide were followed in water and in the worms. In water, fenhexamid concentration decreased slowly (maximum -2 +/- 0.03% after 2 days for 1 mg L(-1)). In the worms, it increased after 4 days and decreased thereafter, confirming that the worms were exposed to the fungicide and not to a degradation product. LC50 values were between 95.22 +/- 5.36 and 32.11 +/- 1.8 mg L(-1) depending on exposure time. Exposure to fenhexamid had a negative effect on T. tubifex growth (maximum effect -12.2 +/- 0.8% after 7 days with 10 mg L(-1)) demonstrating the toxic effect of the pesticide. This growth rate decrease was accompanied by a reduction in protein and glycogen contents. The activity of catalase (CAT), and glutathione reductase (GR) increased in response to the fungicide demonstrating an oxidative stress in the worms. In contrast glutathion-S-transferase activity (GST) decreased. Exposure to fenhexamid also induced synthesis of MT (maximum +78 +/- 8% after 2 days for 10 mg L(-1)). The specificity of MT concentration increase in response to metals is discussed.     

Survival, growth and reproduction of Daphnia carinata (Crustacea: Cladocera) exposed to chronic cadmium stress at different food (Chlorella) levels

In nature, organisms have to respond to a diversity of factors acting simultaneously. The present investigation was conducted to study whether changes in food (Chlorella) levels could modify the chronic toxicity of cadmium on the various life-history parameters, such as survivorship, longevity, life expectancy, fecundity, age at first reproduction, R(0), T, r and growth rates of the cladoceran Daphnia carinata. The study indicated that at low food levels (0.5 x 10(6) cells ml(-1) Chlorella), cadmium concentrations in the range of 27-162 microg litre(-1) reduced these life-history parameters by 50% (EC(50)). At medium food levels (1.5 x 10(6) ml(-1) Chlorella) the EC(50) of cadmium was in the range of 51-127 microg litre(-1). At high food levels (4.5 x 10(6) cells ml(-1) Chlorella), the toxic effect of cadmium was greatly reduced. The decreases in survival, growth and reproduction of D. carinata at high cadmium-low food levels affected the fitness parameter 'r'. The study emphasises the need to include reproductive parameters other than mere survival in toxicity bioassays. The study also stresses the need to incorporate in laboratory tests other relevant factors that might modify pollutant toxicity.     

Low hepatic 7-ethoxyresorufin-O-deethylase (EROD) activity and minor alterations in retinoid and thyroid hormone levels in flounder (Platichthys flesus) exposed to the polychlorinated biphenyl (PCB) mixture, Clophen A50

The effect of the polychlorinated biphenyl (PCB) mixture Clophen A50 on hepatic cytochrome P4501A1 dependent EROD (7-ethoxyresorufin-O-deethylase) activity, plasma thyroid hormone levels and plasma, kidney and liver retinoid concentrations of the euryhaline flatfish flounder (Platichthys flesus) was determined 2 and 10 days after i.p. (intraperitoneal) injection with 20, 100 and 500 mg Clophen A50/kg body weight. No effect of Clophen A50 on total cytochrome P450 content in flounder liver was observed at both time points. A six-fold, dose-dependent, significant increase in EROD activity was found at exposure day 10 in flounder receiving 100 or 500 mg Clophen A50/kg body weight. Plasma retinol concentrations were not altered at both time points after Clophen A50 administration, whereas renal retinol levels showed a minor dose-related increase at day 2 and day 10 of exposure. Significant alterations in hepatic retinoid concentrations were observed, which were not dependent on the dose of PCB administered. In addition Clophen A50 administration did not result in a dose-related alteration of total T4 concentrations in plasma. Total T3 concentrations in plasma were only significantly increased at day 2 after exposure, whereas free T4 concentrations were increased at both time points after Clophen A50 administration. These data indicate that with regard to the parameters investigated and in contrast to other fish species studied, the flounder is not a sensitive species to PCB exposure.