Hans-Christian Teien

Effects of combined γ-irradiation and metal (Al+Cd) exposures in Atlantic salmon (Salmo salar L.).

These experiments were designed to investigate transcriptional effects in Atlantic salmon (Salmo salar) after exposure in vivo to ionizing gamma radiation combined with subtoxic levels of aluminum (Al) and cadmium (Cd). Juvenile fish (35 g) in freshwater with or without Al and Cd (255 microg Al/L + 6 microg Cd/L) were exposed to a 75 mGy dose of gamma-irradiation, and induced responses were compared to those of controls. The transcriptional levels of eight genes encoding proteins known to respond to stress in fish were quantified in liver of fish exposed for 5 h to gamma radiation, to Al and Cd or to the combination of Al, Cd and gamma radiation. The studied genes were caspase 3B, caspase 6A, caspase 7, p53 (apoptosis), glutathione reductase (GR), phospholipid hydroperoxide glutathione peroxidase (GSH-Px), (oxidative stress), metallothionein (MT-A) (metal stress) and ubiquitin (Ubi) (protein degradation). The results showed that gamma-irradiation alone induced significant upregulation of caspase 6A, GR, GSH-Px, MT-A and Ubi compared to the control group, while 5 h exposure to Al+Cd alone did not induce any of the studied genes compared to the control. No significant upregulation of the series of investigated genes could be observed in fish exposed to gamma-irradiation in combination with Al+Cl. In conclusion, the results suggest that the presence of Al+Cd in the water counteracted the gamma-irradiation effect by modifying the transcription of genes encoding proteins involved in the defense mechanisms against free radicals in the cells.

Early stress responses in Atlantic salmon (Salmo salar) exposed to environmentally relevant concentrations of uranium

Uranium (U) is a naturally occurring heavy metal widely used in many military and civil applications. Uranium contamination and the associated potential adverse effects of U on the aquatic environment have been debated during recent years. In order to understand the effect and mode of action (MoA) of U in vivo, juvenile Atlantic salmon (Salmo salar) were exposed to 0.25 mg/L, 0.5 mg/L and 1.0mg/L waterborne depleted uranyl acetate, respectively, in a static system for 48 h. The U concentrations in the gill and liver were analyzed by inductively coupled plasma mass spectrometry (ICP-MS) and the resulting biological effects were determined by a combination of analysis of gene expression and micronuclei formation. The hepatic transcriptional level of 12 biomarker genes from four stress-response categories, including oxidative stress (γ-glutamyl cysteine synthetase (GCS), glutathione reductase (GR), glutathione peroxidase (GPx)), DNA damage and repair (P53, cyclin-dependent kinase inhibitor 1 (P21), growth arrest and DNA damage-inducible gene gamma (Gadd45G), proliferating cell nuclear antigen (PCNA), Rad51), apoptosis (Bcl2-associated X protein (BAX), Bcl-x, Caspase 6A,) and protein degradation (Ubiquitin) were evaluated by quantitative real-time polymerase chain reaction (q-rtPCR). The results clearly showed accumulation of U in the gill and liver with increasing concentrations of U in the exposure water. The effects of U on differential hepatic gene expression also occurred in a concentration-dependent manner, although deviations from ideal concentration-response relationships were observed at the highest U concentration (1.0 mg/L). All the genes tested were found to be up-regulated by U while no significant micronuclei formation was identified. The results suggest that U may cause oxidative stress in fish liver at concentrations greater than 0.25 mg/L, giving rise to clear induction of several toxicologically relevant biomarker genes, although no significant adverse effects were observed after the relatively short exposure period.

Speciation of selected trace elements in three Ethiopian Rift Valley Lakes (Koka, Ziway, and Awassa) and their major inflows.

The Ethiopian Rift Valley Lakes (ERVLs) are water resources which have considerable environmental, economic and cultural importance. However, there is an increasing concern that increasing human activities around these lakes and their main inflows can result in increased contamination of these water bodies. Information on total concentrations of some trace elements is available for these lakes and their inflows; however, data on the trace element speciation is lacking. Therefore, the objective of this study was to determine the low molecular mass (LMM) trace element species and also, evaluate the influence of flooding episodes on the LMM trace element fractions. At-site size and charge fractionation system was used for sampling of water from the lakes Koka, Ziway and Awassa and their main inflows during the dry and wet seasons. The results showed that chromium (Cr), manganese (Mn), cobalt (Co), nickel (Ni), copper (Cu), zinc (Zn), and lead (Pb) in Lake Koka and its inflows as well as in Lake Ziway were predominantly present as HMM (high molecular mass, i.e., >10 kDa) forms, while arsenic (As), selenium (Se), cadmium (Cd) were more mobile during the dry season. In Lake Awassa, all except Cr and Mn were predominantly found as LMM species (low molecular mass, i.e. <10 kDa) which can be attributed to the high concentrations of LMM DOC (dissolved organic carbon). During the wet season, results from the Lake Koka and its inflows showed that all trace elements were predominantly associated with HMM forms such as colloids and particles, demonstrating that the mobility of elements was reduced during the wet season. The colloidal fraction of elements such as Cr, Ni, and Cd was also correlated with dissolved Fe. As the concentration of LMM trace element species are very low, the mobility, biological uptake and the potential environmental impact should be low.

Toxicity of road deicing salt (NaCl) and copper (Cu) to fertilization and early developmental stages of Atlantic salmon (Salmo salar)

In many countries, salting of ice or snow covered roads may affect aquatic organisms in the catchment directly or indirectly by mobilization of toxic metals. We studied the toxicity of road deicing salt and copper (Cu) on the vulnerable early life stages of Atlantic salmon (Salmo salar), from fertilization till hatching. Controlled episodic exposure to road salt (≥ 5,000 mg/L) during fertilization resulted in reduced swelling and less percent egg survival. Exposure to Cu both during and post fertilization caused delayed hatching. Larval deformities were, however found as an additional effect, when eggs were exposed to high salt concentration (≥ 5,000 mg/L) mixed with Cu (10 μg Cu/L) during fertilization. Thus, it appears that the sensitivity of early developmental stages of Atlantic salmon increased when exposed to these stressors, and road salt application during spawning can pose threat to Atlantic salmon in water bodies receiving road runoff. The study gives insight on assessment and management of risks on Atlantic salmon population posed by road related hazardous chemicals.

Risk of acute toxicity for fish during aluminium application to hardwater lakes

To assess the risk of aluminium (Al) toxicity during the restoration of the eutrophic lake Tiefwarensee by hypolimnetic addition of NaAl(OH)(4)-solution (aluminate) the generally limnological monitoring was accompanied by fractionation of Al in water and using Al accumulation on fish gills as bioindicator. The concentration of reactive Al species in the alkaline water (pH 8) peaked at 2mgL(-1) in parts of the anoxic hypolimnion and was 0.088+/-0.053mgL(-1) (n=70) in the epilimnion during the five years of treatment. During an Al treatment cycle in summer 2003, perches showed significant Al accumulation on gills (100microg Al g(-1) dw) whereas roaches, breams and silver carps remained unaffected. Thus, the Al toxicity towards several fish species seems to be low, although the concentration of reactive Al in the lake water increased by a factor of 2. However, high Al toxicity due to lake treatment with aluminate could not be excluded, as high Al-gill concentration was observed. An Al balance two years after the treatment indicates complete export of the added Al into the sediment.

Hepatic transcriptomic profiling reveals early toxicological mechanisms of uranium in Atlantic salmon (Salmo salar)

BackgroundUranium (U) is a naturally occurring radionuclide that has been found in the aquatic environment due to anthropogenic activities. Exposure to U may pose risk to aquatic organisms due to its radiological and chemical toxicity. The present study aimed to characterize the chemical toxicity of U in Atlantic salmon (Salmo salar) using depleted uranium (DU) as a test model. The fish were exposed to three environmentally relevant concentrations of DU (0.25, 0.5 and 1.0 mg U/L) for 48 h. Hepatic transcriptional responses were studied using microarrays in combination with quantitative real-time reverse transcription polymerase chain reaction (qPCR). Plasma variables and chromosomal damages were also studied to link transcriptional responses to potential physiological changes at higher levels.ResultsThe microarray gene expression analysis identified 847, 891 and 766 differentially expressed genes (DEGs) in the liver of salmon after 48 h exposure to 0.25, 0.5 and 1.0 mg/L DU, respectively. These DEGs were associated with known gene ontology functions such as generation of precursor metabolites and energy, carbohydrate metabolic process and cellular homeostasis. The salmon DEGs were then mapped to mammalian orthologs and subjected to protein-protein network and pathway analysis. The results showed that various toxicity pathways involved in mitochondrial functions, oxidative stress, nuclear receptor signaling, organ damage were commonly affected by all DU concentrations. Eight genes representative of several key pathways were further verified using qPCR No significant formation of micronuclei in the red blood cells or alterations of plasma stress variables were identified.ConclusionThe current study suggested that the mitochondrion may be a key target of U chemical toxicity in salmon. The induction of oxidative stress and uncoupling of oxidative phosphorylation may be two potential modes of action (MoA) of DU. These MoAs may subsequently lead to downstream events such as apoptosis, DNA repair, hypoxia signaling and immune response. The early toxicological mechanisms of U chemical toxicity in salmon has for the first time been systematically profiled. However, no other physiological changes were observed. Future efforts to link transcriptional responses to adverse effects have been outlined as important for understanding of potential risk to aquatic organisms.

Fish mortality during sea salt episodes—catchment liming as a countermeasure

Aluminium (Al) toxicity is usually associated with acid rain and acidified freshwater systems. The present work demonstrates that acute fish mortality (50%) also occurs in moderate acidified salmon rivers during sea salt episodes. Furthermore, catchment liming was proved to be an efficient measure to counteract the fish toxicity. The impact of sea salt episodes on river water qualities and on Atlantic Salmon (Salmo salar L.) was studied in two rivers situated at the west coast of Norway. During February-May 2002, fish were kept in tanks and continually exposed to the changing water qualities. Changes in Al-species were followed using in situ fractionation techniques. During storm events and high sea salt deposition, the sea salt concentration increased (190 to 580 microM Cl), pH decreased (pH 5.3 to 4.6) and the concentration of low molecular mass (LMM) cationic Al-species (Al(i)) increased (0.7 to 3.0 microM) in the river. Subsequently, Al accumulated in fish gills (6 to 19 micromol g(-1) dw) causing ionoregulatory and respiratory failures as well as mortality. In water the concentration of LMM Al(i) stayed enhanced during four weeks, while the physiological stress responses in surviving fish remained high for a longer time (>eight weeks). To counteract Al toxicity, one of the tributary catchments had been limed four years earlier. Due to catchment liming (1000 kg ha(-1)) the water concentration of LMM Al(i)(<0.7 microM) and the Al accumulation in gills remained relatively low (<7 micromol g(-1) dw) during the storm and no fish mortality occurred.

Aluminum exposure impacts brain plasticity and behavior in Atlantic salmon (Salmo salar).

SUMMARY Aluminum (Al) toxicity occurs frequently in natural aquatic ecosystems as a result of acid deposition and natural weathering processes. Detrimental effects of Al toxicity on aquatic organisms are well known and can have consequences for survival. Fish exposed to Al in low pH waters will experience physiological and neuroendocrine changes that disrupt homeostasis and alter behavior. To investigate the effects of Al exposure on both the brain and behavior, Atlantic salmon (Salmo salar) kept in water treated with Al (pH 5.7, 0.37±0.04 μmol 1−1 Al) for 2 weeks were compared with fish kept in under control conditions (pH 6.7, <0.04 μmol 1−1 Al). Fish exposed to Al and acidic conditions had increased Al accumulation in the gills and decreased gill Na+, K+-ATPase activity, which impaired osmoregulatory capacity and caused physiological stress, indicated by elevated plasma cortisol and glucose levels. Here we show for the first time that exposure to Al in acidic conditions also impaired learning performance in a maze task. Al toxicity also reduced the expression of NeuroD1 transcript levels in the forebrain of exposed fish. As in mammals, these data show that exposure to chronic stress, such as acidified Al, can reduce neural plasticity during behavioral challenges in salmon, and may impair the ability to cope with new environments.

Main results of the 2012 joint Norwegian–Russian expedition to the dumping sites of the nuclear submarine K-27 and solid radioactive waste in Stepovogo Fjord, Novaya Zemlya

This paper reports the main results of the 2012 joint Norwegian-Russian expedition to investigate the radioecological situation of the Stepovogo Fjord on the eastern coast of Novaya Zemlya, where the nuclear submarine K-27 and solid radioactive waste was dumped. Based on in situ gamma measurements and the analysis of seawater and sediment samples taken around the submarine, there was no indication of any leakage from the reactor units of K-27. With regard to the radioecological status of Stepovogo Fjord, activity concentrations of all radionuclides in seawater, sediment and biota in 2012 were in general lower than reported from the previous investigations in the 1990s. However in 2012, the activity concentrations of (137)Cs and, to a lesser extent, those of (90)Sr remained elevated in bottom water from the inner part of Stepovogo Fjord compared with surface water and the outer part of Stepovogo Fjord. Deviations from expected (238)Pu/(239,240)Pu activity ratios and (240)Pu/(239)Pu atom ratios in some sediment samples from the inner part of Stepovogo Fjord observed in this study and earlier studies may indicate the possibility of leakages from dumped waste from different nuclear sources. Although the current environmental levels of radionuclides in Stepovogo Fjord are not of immediate cause for concern, further monitoring of the situation is warranted.

Dose-dependent hepatic transcriptional responses in Atlantic salmon (Salmo salar) exposed to sublethal doses of gamma radiation

Due to the production of free radicals, gamma radiation may pose a hazard to living organisms. The high-dose radiation effects have been extensively studied, whereas the ecotoxicity data on low-dose gamma radiation is still limited. The present study was therefore performed using Atlantic salmon (Salmo salar) to characterize effects of low-dose (15, 70 and 280 mGy) gamma radiation after short-term (48h) exposure. Global transcriptional changes were studied using a combination of high-density oligonucleotide microarrays and quantitative real-time reverse transcription polymerase chain reaction (qPCR). Differentially expressed genes (DEGs; in this article the phrase gene expression is taken as a synonym of gene transcription, although it is acknowledged that gene expression can also be regulated, e.g., at protein stability and translational level) were determined and linked to their biological meanings predicted using both Gene Ontology (GO) and mammalian ortholog-based functional analyses. The plasma glucose level was also measured as a general stress biomarker at the organism level. Results from the microarray analysis revealed a dose-dependent pattern of global transcriptional responses, with 222, 495 and 909 DEGs regulated by 15, 70 and 280 mGy gamma radiation, respectively. Among these DEGs, only 34 were commonly regulated by all radiation doses, whereas the majority of differences were dose-specific. No GO functions were identified at low or medium doses, but repression of DEGs associated with GO functions such as DNA replication, cell cycle regulation and response to reactive oxygen species (ROS) were observed after 280mGy gamma exposure. Ortholog-based toxicity pathway analysis further showed that 15mGy radiation affected DEGs associated with cellular signaling and immune response; 70mGy radiation affected cell cycle regulation and DNA damage repair, cellular energy production; and 280mGy radiation affected pathways related to cell cycle regulation and DNA repair, mitochondrial dysfunction and immune functions. Twelve genes representative of key pathways found in this study were verified by qPCR. Potential common MoAs of low-dose gamma radiation may include induction of oxidative stress, DNA damage and disturbance of oxidative phosphorylation (OXPHOS). Although common MoAs were proposed, a number of DEGs and pathways were still found to be dose-specific, potentially indicating multiple mechanisms of action (MOAs) of low-dose gamma radiation in fish. In addition, plasma glucose displayed an apparent increase with increasing radiation doses, although the results were not significantly different from the control. These findings suggested that sublethal doses of gamma radiation may cause dose-dependent transcriptional changes in the liver of Atlantic salmon after short-term exposure. The current study predicted multiple MoA for gamma radiation and may aid future impact assessment of environmental radioactivity in fish.