Bioaccumulation of trace elements in liver and kidney of fish species from three freshwater lakes in the Ethiopian Rift Valley.

The objective of the present work was to obtain scientific information on the ecological health of three freshwater lakes (Awassa, Koka, and Ziway) situated in the Ethiopian Rift Valley by investigating possible trace element contamination accumulated in fish. Accordingly, fish liver and kidney samples were collected from three commercially important fish species (Barbus intermedius, Clarias gariepinus, and Oreochromis niloticus) in the lakes to determine the concentrations of chromium (Cr), manganese (Mn), cobalt (Co), nickel (Ni), copper (Cu), zinc (Zn), arsenic (As), selenium (Se), cadmium (Cd), and lead (Pb), using ICP-MS. Trace element concentrations were generally higher in O. niloticus compared with concentrations in B. intermedius and C. gariepinus. Compared to background values of most freshwater fish species, higher liver concentrations of Cu in C. gariepinus and O. niloticus, Mn in O. niloticus, Co in all except B. intermedius, and Zn in C. gariepinus from Lakes Ziway and Awassa were found. Cr, Co, Ni, Cd, and Pb were enriched in kidney, while Mn, Cu, Zn, As, and Se seems retained in the liver tissues. Assessment of transfer factors indicated that bioaccumulation from water and diet occurred, while uptake from sediments was low. Furthermore, the transfer factor values were generally higher for essential elements compared to the non-essential elements. Multivariate statistical analyses showed that the differences between the trace element levels were generally not significant among the lakes (p = 0.672), while significant differences were found between the fish species (p = 0.042), and between accumulation in kidney and liver (p = 0.002).

Single and multiple stressor effect of road deicers and Cu on Atlantic salmon (Salmo salar) alevins from hatching till swim-up.

Road salts are frequently used for deicing of roads in the Nordic countries. During snow-melt, the road run-off containing high concentrations of road salt and various metals such as Cu remobilized from sand, silt and dust may negatively influence organisms in downstream receiving water bodies. The present work focuses on the impact of road salt (NaCl) and Cu, separately and in mixtures on Atlantic salmon alevins from hatching till swim-up. The results showed that high road salt concentrations could induce a series of negative effects in alevins such as reduced growth, deformities, delayed swim-up and mortality. For alevins exposed to all tested road salt concentrations (100-1000mg/L), mortality was significantly higher compared to control. In exposure to Cu solutions (5-20µgCu/L), no effects on growth, morphology, swim-up or mortality of alevins compared to control were observed. In mixture solutions (road salt and Cu), ultrafiltration of the exposure water demonstrated that only 20%-40% of Cu was present as positively charged low molecular mass (LMM) Cu species assumed to be bioavailable. When exposed to road salt and Cu mixtures, negative effects in alevins such as reduced growth, deformities, delayed swim-up and mortality were observed. The overall results indicated that the road salt application could seriously affect sensitive life stages of Atlantic salmon, and application of road salt should be avoided during the late winter-early spring period.

Accumulation of lead (Pb) in brown trout (Salmo trutta) from a lake downstream a former shooting range.

An environmental survey was performed in Lake Kyrtjønn, a small lake within an abandoned shooting range in the south of Norway. In Lake Kyrtjønn the total water concentrations of Pb (14µg/L), Cu (6.1µg/L) and Sb (1.3µg/L) were elevated compared to the nearby reference Lake Stitjønn, where the total concentrations of Pb, Cu and Sb were 0.76, 1.8 and 0.12µg/L, respectively. Brown trout (Salmo trutta) from Lake Kyrtjønn had very high levels of Pb in bone (104mg/kg w.w.), kidney (161mg/kg w.w.) and the gills (137mg/kg d.w), and a strong inhibition of the ALA-D enzyme activity were observed in the blood (24% of control). Dry fertilized brown trout eggs were placed in the small outlet streams from Lake Kyrtjønn and the reference lake for 6 months, and the concentrations of Pb and Cu in eggs from the Lake Kyrtjønn stream were significantly higher than in eggs from the reference. More than 90% of Pb accumulated in the egg shell, whereas more than 80% of the Cu and Zn accumulated in the egg interior. Pb in the lake sediments was elevated in the upper 2-5cm layer (410-2700mg/kg d.w), and was predominantly associated with redox sensitive fractions (e.g., organic materials, hydroxides) indicating low potential mobility and bioavailability of the deposited Pb. Only minor amounts of Cu and Sb were deposited in the sediments. The present work showed that the adult brown trout, as well as fertilized eggs and alevins, may be subjected to increased stress due to chronic exposure to Pb, whereas exposure to Cu, Zn and Sb were of less importance.

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

BACKGROUND: Uranium (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. RESULTS: The 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. CONCLUSION: The 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.

Organochlorine pesticides and polychlorinated biphenyls in fish from Lake Awassa in the Ethiopian Rift Valley: human health risks.

Dietary intake of fish containing organic contaminants poses a potential threat to human health. In the present work, an assessment has been carried out to look at the human health risk associated with consumption of fish contaminated with organochlorine pesticides (OCPs) and polychlorinated biphenyles (PCBs) in certain fish species collected from Lake Hawassa, Ethiopia. The health risk assessment was made by comparing the concentrations of OCPs and PCBs in fish muscle tissues with reference doses given in the USEPA guidelines. Dichlorodiphenyltrichloroethanes (DDTs), endosulfans, PCBs and chloridanes were identified in fish species collected from Lake Hawassa. The most predominant pesticides were DDTs, with mean concentrations of ΣDDT ranging from 19 to 56 ng g(-1) wet weights. The highest concentrations of DDTs were found in Barbus intermedius, representing the highest trophic level. PCBs, DDT and endosulfan concentrations found in B. intermedius exceeded the reference dose for children between the ages of 0-1 year (with hazard index of above 1.0). Therefore, consumption of fish from a high trophic level (e.g. B. intermedius) from Lake Hawassa may pose a special health risk to children.

Biomagnification of DDT and its metabolites in four fish species of a tropical lake.

The concentrations and biomagnifications of dichlorodiphenyltrichloroethane (DDT) and its metabolites were examined in four fish species (Clarias gariepinus, Oreochromis niloticus, Tilapia zillii, and Carassius auratus) from Lake Ziway, Rift Valley, Ethiopia. Paired stomach content analysis, and stable isotope ratio of nitrogen (δ(15)N, ‰) and carbon (δ(13)C, ‰) were used to study the trophic position of the fish species in the lake. 4,4'-DDE, 4,4'-DDT and 4,4'-DDD were the main DDTs identified in the fish samples, with 4,4'-DDE as the most predominant metabolite, with mean concentration ranging from 1.4 to 17.8 ng g(-1) wet weight (ww). The concentrations of DDTs found in fish from Lake Ziway were, in general lower than those found in most studies carried out in other African Lakes. However, the presence of DDT in all tissue samples collected from all fish species in the lake indicates the magnitude of the incidence. Moreover, the observed mean 4,4'-DDE to 4,4'-DDT ratio below 1 in C. auratus from Lake Ziway may suggest a recent exposure of these species to DDT, indicating that a contamination source is still present. 4,4'-DDE was found to biomagnify in the fish species of the lake, and increases with trophic level, however, the biomagnification rate was generally lower than what has been reported from other areas. Significantly higher concentrations of 4,4'-DDE were found in the top consumer fish in Lake Ziway, C. gariepinus than in O. niloticus (t=2.6, P<0.01), T. zillii (t=2.5, P<0.02) and C. auratus (t=2.2, P<0.03).

Mercury concentrations in commercial fish species of Lake Phewa, Nepal.

Mercury (Hg) concentrations in four commercial fish species (Tilapia Oreochromis niloticus, Spiny Eel Mastacembelus armatus, African catfish Clarias gariepinus, and Sahar Tor putitora), were investigated in Lake Phewa, Nepal. Mean values of total mercury (THg mg kg(-1), ww) in these fishes were 0.02, 0.07, 0.05, and 0.12 respectively. Methylmercury contributed 82 % of THg. The lowest value was detected in O. niloticus, an exclusive plant feeder. The biomagnification rate of Hg through the fish community was 0.041 per δ(15)N (‰). The present investigation produced an important baseline data of Hg pollution in the fish community in this region.

Pre-anaesthetic metomidate sedation delays the stress response after caudal artery cannulation in Atlantic cod (Gadus morhua).

Recovery from caudal artery cannulation with and without pre-anaesthesia metomidate sedation was assessed in Atlantic cod (Gadus morhua). The levels of plasma cortisol, glucose, electrolytes and acid-base parameters were compared between sedated and unsedated cod and to those in uncannulated individuals, where the samples were obtained by sacrificial sampling (reference level). Metomidate sedation delayed the stress response, causing sedated cod plasma cortisol to return to the reference level more slowly [day 4 post surgery (PS)] than in unsedated cod (day 2 PS). Plasma glucose was elevated in both sedated and unsedated cod up to and including day 5 PS. Plasma K(+) was lower and pH was higher in cannulated cod than in the reference from 24 h PS until the end of experimentation, indicating a stress effect of sacrificial sampling on plasma K(+) and pH that was likely caused by an acute stress response. Metomidate sedation delayed the stress response following CA cannulation and should therefore not be used as a pre-anaesthetic sedation in Atlantic cod. The caudal artery cannulation can be a useful tool in obtaining repeated blood samples from Atlantic cod given an adequate recovery time, which was determined to be 6 days irrespective of pre-anaesthesia sedation status.

Mercury concentrations are low in commercial fish species of Lake Ziway, Ethiopia, but stable isotope data indicated biomagnification.

Stable isotope ratios of nitrogen (δ(15)N) and carbon (δ(13)C), complemented by stomach content data, were used to assess the food web structure and trophic transfer of mercury (Hg) in four commercial fish species of Lake Ziway, Ethiopia: Nile tilapia (Oreochromis niloticus), catfish (Clarias gariepinus), Tilapia zillii, and golden carp (Carassius auratus). Total mercury (THg in mg kg(-1), ww) concentrations were low, with mean values of 0.033, 0.034, 0.025, and 0.011, in C. gariepinus, T. zillii, C. auratus, and O. niloticus, respectively. The relationships between mercury concentrations against total length (TL) and total weight (TW) were positive and significant in T. zillii, C. auratus, and C. gariepinus (P<0.01), but not in O. niloticus, which even showed a decreasing tendency with increase in TL and TW. Regression of log THg vs. δ(15)N among all fish species showed a significant correlation, indicating that mercury is biomagnifying along the food web of Lake Ziway. Isotope ratios indicated that C. gariepinus occupied the highest trophic level of the food web of Lake Ziway; but contained similar THg concentrations as T. zillii, which is located at a lower trophic level, probably due to a faster growth rate of C. gariepinus, and thus an example of biodilution.

Changes in arterial PO2, physiological blood parameters and intracellular antioxidants in free-swimming Atlantic cod (Gadus morhua) exposed to varying levels of hyperoxia.

Free-swimming Atlantic cod (Gadus morhua) were exposed to water oxygen pressures (P(w)O2) ranging from 18.1 to 41.5 kPa and sampled for blood using an indwelling caudal artery cannula. Arterial blood oxygen pressure (P(a)O2) increased with increasing P(w)O2, from 12.0 kPa in normoxia (18.1 kPa) to 34.2 kPa in the highest hyperoxic level tested (41.5 kPa). Blood CO2 pressure and plasma bicarbonate concentration increased with P(w)O2, indicating reduced ventilation with increased P(w)O2. Plasma glucose, sodium and potassium were not affected by water oxygen level. Blood oxidative stress biomarkers, reduced glutathione, oxidized glutathione and the oxidative stress index (ratio between oxidized and total glutathione) differed intermittently between normoxia and hyperoxia. The oxidative stress index was higher in the blood of exposed compared to unexposed control cod. Together with elevated P(a)O2, these findings suggest increased production of reactive oxygen species and increased oxidative stress in Atlantic cod exposed to hyperoxia.

The legacy from the 50 years of acid rain research, forming present and future research and monitoring of ecosystem impact : This article belongs to Ambio's 50th Anniversary Collection. Theme: Acidification.

Acid rain and acidification research are indeed a multidisciplinary field. This field evolved from the first attempts to mitigate acid freshwater in the 1920s, then linking acid rain to the acidification in late 1950s, to the broad project-concepts on cause and effect from the late 1960s. Three papers from 1974, 1976 and 1988 demonstrate a broad approach and comprise scientific areas from analytical chemistry, biochemistry, limnology, ecology, physiology and genetics. Few, if any, environmental problems have led to a public awareness, political decisions and binding limitations as the story of acid rain. Acid precipitation and acidification problems still exist, but at a lower pressure, and liming has been reduced accordingly. However, the biological responses in the process of recovery are slow and delayed. The need for basic science, multidisciplinary studies, long time series of high-quality data, is a legacy from the acid rain era, and must form the platform for all future environmental projects.

Ecotoxicological impact of highway runoff using brown trout (Salmo trutta L.) as an indicator model.

The ecotoxicological impact of highway runoff on brown trout (Salmo trutta L.) was studied in an in situ experiment consisting of four 24 h simulated runoff episodes. Fish were maintained in 5 tanks and exposed to highway runoff from a sedimentation pond close to E6 outside the city of Oslo, Norway. The tanks had the following contaminant loadings during the episodes: stream water (control), pond inlet, pond outlet, pond inlet + stream water and pond outlet + stream water. Opposite to road salt and compared to earlier findings, the first two episodes had rather low concentrations of trace metals, hydrocarbons and polycyclic aromatic hydrocarbons. A heavy rainfall before episode 3 increased the concentrations of all the contaminants except road salt which was diluted. In addition, lowered oxygen levels led to hypoxic conditions. Overall the fish exposed to highway runoff had, compared to the control fish, higher concentrations of trace metals in gills and liver, increased activity of the antioxidant defense system represented by superoxide dismutase, catalase and metallothionein, problems with the regulation of plasma Cl and Na, as well as increased levels of blood glucose and pCO(2). Finally, this seemed to affect the metabolism of the fish through reduced condition factor. The observed effects were likely caused by multiple stressors and not by a single contaminant. The sedimentation pond clearly reduced the toxicity of the highway runoff. But even in the least polluted exposure tank (pond outlet + stream water) signs of physiological disturbances were evident.

Selective exploitation of large pike Esox lucius--effects on mercury concentrations in fish populations.

The present study outlines two main trends of mercury transfer patterns through the fish community: 1) the Hg concentrations increase with increase in the trophic level, with top predators having the highest concentrations, and 2) a fast growth rate may dilute the concentrations of Hg in fish muscle tissue (growth biodilution). In 2004, an extensive reduction in number of large pike (Esox lucius L.), was initiated by selective gillnet fishing in Lake Arungen, Norway, in order to increase the pike recruitment due to an expected reduction in cannibalism. In this connection, total mercury (THg) concentrations in the fish community were studied both before (2003) and after (2005) the removal of large pike. The delta(15)N signatures and stomach content analyses indicated that pike and perch (Perca fluviatilis L.) occupied the highest trophic position, while roach (Rutilus rutilus (L.)) was at the lower level, and rudd (Scardinius erythrophthalmus L.) at the lowest. The piscivores, pike and perch, had the highest concentrations of THg. The biomagnification rate of THg through the food web in the fish community was 0.163 (per thousand delta(15)N), with the highest uptake rate (0.232) in perch. A significant decrease in THg concentrations was found in all fish species in 2005 compared to 2003. Removal of the top predators in an Hg contaminated lake might thus be an important management tool for reducing Hg levels in fish, thereby reducing health risk to humans.

Route of exposure has a major impact on uptake of silver nanoparticles in Atlantic salmon (Salmo salar).

The potential impact of silver nanoparticles (Ag NPs) on aquatic organisms is to a large extent determined by their bioavailability through different routes of exposure. In the present study juvenile Atlantic salmon (Salmo salar) were exposed to different sources of radiolabeled Ag (radiolabeled 110m Ag NPs and 110m AgNO3 ). After 48 h of waterborne exposure to 3 µg/L citrate stabilized 110m Ag NPs or 110m AgNO3 , or a dietary exposure to 0.6 mg Ag/kg fish (given as citrate stabilized or uncoated 110m Ag NPs, or 110m AgNO3 ), Ag had been taken up in fish regardless of route of exposure or source of Ag (Ag NPs or AgNO3 ). Waterborne exposure led to high Ag concentrations on the gills, and dietary exposure led to high concentrations in the gastrointestinal tract. Silver distribution to the target organs was similar for both dietary and waterborne exposure, with the liver as the main target organ. The accumulation level of Ag was 2 to 3 times higher for AgNO3 than for Ag NPs when exposure was through water, whereas no significant differences were seen after dietary exposure. The transfer (Bq/g liver/g food or water) from exposure through water was 4 orders of magnitude higher than from feed using the smallest, citrate-stabilized Ag NPs (4 nm). The smallest NPs had a 5 times higher bioavailability in food compared with the larger and uncoated Ag NPs (20 nm). Despite the relatively low transfer of Ag from diet to fish, the short lifetime of Ag NPs in water and their transfer to sediment, feed, or sediment-dwelling food sources such as larvae and worms could make diet a significant long-term exposure route. Environ Toxicol Chem 2018;37:2895-2903. © 2018 SETAC.

Uptake and effects of 2, 4, 6 - trinitrotoluene (TNT) in juvenile Atlantic salmon (Salmo salar).

Organ specific uptake and depuration, and biological effects in Atlantic salmon (Salmo salar) exposed to 2, 4, 6-trinitrotoluene (TNT) were studied. Two experiments were conducted, the first using radiolabeled TNT (14C-TNT, 0.16mg/L) to study uptake (48h) and depuration (48h), while the second experiment focused on physiological effects in fish exposed to increasing concentrations of unlabeled TNT (1µg-1mg/L) for 48h. The uptake of 14C-TNT in the gills and most of the organs increased rapidly during the first 6h of exposure (12h in the brain) followed by a rapid decrease even though the fish were still exposed to TNT in the water. The radioactivity in the gall bladder reached a maximum after 55h, 7h after the transfer to the clean water. A high concentration of 14C-TNT in the gall bladder indicates that TNT is excreted through the gall bladder. Mortality (2 out of 14) was observed at a concentration of 1mg/L, and the surviving fish had hemorrhages in the dorsal muscle tissue near the spine. Analysis of the physiological parameters in blood from the high exposure group revealed severe effects, with an increase in the levels of glucose, urea and HCO3, and a decrease in hematocrit and the levels of Cl and hemoglobin. No effects on blood physiology were observed in fish exposed to the lower concentrations of TNT (1-100µg/L). TNT and the metabolites 2-amino-4,6-dinitrotoluene (2-ADNT) and 4-amino-2,6-dinitrotoluene (4-ADNT) were found in the muscle tissue, whereas only 2-ADNT and 4-ADNT were found in the bile. The rapid excretion and estimated bioconcentration factors (range of 2-18 after 48h in gills, blood, liver, kidney, muscle and brain) indicated a low potential for bioaccumulation of TNT.

Lower than expected mercury concentration in piscivorous African sharptooth catfish Clarias gariepinus (Burchell).

The concentrations of total mercury (THg), stable isotopes of nitrogen (delta(15)N) and carbon (delta(13)C), and the diet of the African sharptooth catfish Clarias gariepinus in Lake Awassa, Ethiopia, were studied from January 2003 to February 2004. Values of the delta(15)N were used as an index of trophic position in four length groups and compared to actual stomach contents. The diet of C. gariepinus within the length range of 201-600 mm L(T) mainly consisted of the small barb Barbus paludinosus, aquatic insects, and mollusks. The proportion of fish prey in the diet was 60% by volume, irrespective of fish size. The differences in delta(15)N values of individuals within and between length classes were less than 3 per thousand, and were not significantly related to total length, showing the similarity in trophic niche of the different sizes, which corresponded to recorded stomach contents. Mercury concentrations were in the range of 0.002-0.154 mg kg(-1) ww, and had no significant relationship to total length. Hence, even large specimens of C. gariepinus have Hg values below the WHO threshold of 0.2 mg kg(-1) ww. The slope of the regression line between log [Hg] and delta(15)N was small, 0.06, indicating the absence of trophic shifts and biomagnification of Hg in larger specimens in our samples. The low Hg concentrations in C. gariepinus compared to the Hg concentrations in other piscivorous fish species in Lake Awassa, such as Barbus intermedius and B. paludinosus, may be due to its dependence on invertebrate preys at small size, diet switching towards low Hg prey fish at larger size, and growth biodilution owing to higher growth rate.

Gill reactivity of aluminium-species following liming.

In acidified aluminium (Al) rich freshwater positively charged Al-species (Al(i)) are the key toxic components due to the accumulation in fish gills. As a countermeasure, liming is used to increase the pH and reduce the concentration of Al(i)-species; in particular low molecular mass (LMM) Al(i)-species by hydrolyses. However, very toxic high molecular mass (HMM) Al polymers can form in the unstable mixing zone immediately after liming. In the present work gill reactivity of LMM and HMM Al-species was studied under controlled conditions in eight channel-tank mixing zone systems in the field where Atlantic salmon (Salmo salar L.) kept in cages were exposed to defined mixing zone water. Mixing zones were created by continually liming acid river water (pH 5.0-5.7) high in LMM Al(i) to pH 6.0 and 6.4, respectively. Transformation processes affecting the Al-speciation as a function of time after liming were documented by in situ hollow fibre ultrafiltration interfaced with ion chromatography, while the Al accumulation in fish gills was used as bioindicator. For fish exposed to mixing zone water immediately after liming (1 min) the Al accumulation in gills (mug Al g(-1)) was higher (factor of 2) than for fish exposed to acid water prior to liming, due to the formation of gill reactive HMM Al(i)-species. The Al accumulation in gills followed a first order kinetic expression reaching steady-state conditions after 24-h exposures. The deposition rate of Al in gills (mug Al g(-1) h(-1)) correlated with the water concentrations of HMM Al(i) (R(2)=0.80) in the mixing zones, and for LMM Al(i) in the acid water (R(2)=0.92). Due to the transient nature of HMM Al(i) the deposition rate of Al decreased from the point of liming with a factor of 10 downstream the channel-tank system (i.e. 100 min after liming). The concentration of gill accumulated Al was higher (factor of 3) immediately after high level liming (pH 6.4) than following low level liming (pH 6.0). However, high level liming was more efficient in detoxifying Al in downstream waters. Furthermore, the bioavailability of a given LMM Al(i) concentration as well as bioreactivity following liming was dependent on the TOC and silicon concentration in the acid water. Increased concentration of TOC (1.5 to 4.3 mg l(-1)) and silicon (0.3 to 1.0 mg l(-1)) reduced the gill reactivity of Al(i) by approximately 50%.

Sodium silicate as alternative to liming-reduced aluminium toxicity for Atlantic salmon (Salmo salar L.) in unstable mixing zones.

When acid aluminium (Al) rich water is limed, unstable mixing zones are formed until equilibrium is reached. In such mixing zones transient high molecular mass positively charged Al-species (HMM Al(i)) being extremely gill reactive are produced, causing toxic effects in fish. The transient HMM Al(i)-species are formed due to hydrolysis and polymerization of low molecular positively charged Al-species (LMM Al(i)), e.g. initiated by liming and the subsequent increase in pH. To counteract the toxicity of transient Al polymers in such mixing zones, sodium silicate, forming non-toxic hydroxyaluminosilicate (HAS) complexes, can be used as alternative to liming. In the present work the effect of sodium silicate on polymerization of LMM Al(i) in unstable mixing zones and subsequent gill reactivity and mortality of fish was compared to results obtained from liming. Diluted sodium silicate (<1.5 g l(-1)) and lime slurry (Ca(OH)(2)), respectively, were continually added to acidified Al-rich water in six different channel-tank systems, to obtain mixing zones with pH 5.9, 6.0 and 6.4, respectively. Utilising in situ size and charge fractionation techniques and following the exposure of Atlantic presmolt (Salmo salar L.) kept in cages at defined stations along the channel-tank systems, changes of Al-species in the mixing zones, the gill reactivity of Al-species and thus Al toxicity could be followed downstream the confluences (time of reaction after mixing: 1-100 min). By increasing the pH of the acid water to 6.0 or 6.4 by sodium silicate, the detoxification of Al was faster than using lime. Using sodium silicate, the transformation of LMM Al(i), the formation of HMM Al(i), the Al deposition in fish gills and fish mortality were lower than using lime. The formation of neutral LMM Al-species (Al(o)) was, however, higher and the formation of colloidal Al-species (Al(c)) lower in the presence of silicate compared to lime. Furthermore, the Al deposition in fish gills and fish mortality decreased by increasing concentration of sodium silicate dosed. Thus, sodium silicate is a good alternative to liming, and under certain circumstances when aging of water may represent a problem (e.g. aquaculture) sodium silicate should be the preferred agent.

Hepatic transcriptional responses in Atlantic salmon (Salmo salar) exposed to gamma radiation and depleted uranium singly and in combination.

Radionuclides are a special group of substances posing both radiological and chemical hazards to organisms. As a preliminary approach to understand the combined effects of radionuclides, exposure studies were designed using gamma radiation (Gamma) and depleted uranium (DU) as stressors, representing a combination of radiological (radiation) and chemical (metal) exposure. Juvenile Atlantic salmon (Salmo salar) were exposed to 70mGy external Gamma dose delivered over the first 5h of a 48h period (14mGy/h), 0.25mg/L DU were exposed continuously for 48h and the combination of the two stressors (Combi). Water and tissue concentrations of U were determined to assess the exposure quality and DU bioaccumulation. Hepatic gene expression changes were determined using microarrays in combination with quantitative real-time reverse transcription polymerase chain reaction (qPCR). Effects at the higher physiological levels were determined as plasma glucose (general stress) and hepatic histological changes. The results show that bioaccumulation of DU was observed after both single DU and the combined exposure. Global transcriptional analysis showed that 3122, 2303 and 3460 differentially expressed genes (DEGs) were significantly regulated by exposure to gamma, DU and Combi, respectively. Among these, 349 genes were commonly regulated by all treatments, while the majority was found to be treatment-specific. Functional analysis of DEGs revealed that the stressors displayed similar mode of action (MoA) across treatments such as induction of oxidative stress, DNA damage and disturbance of oxidative phosphorylation, but also stressor-specific mechanisms such as cellular stress and injury, metabolic disorder, programmed cell death, immune response. No changes in plasma glucose level as an indicator of general stress and hepatic histological changes were observed. Although no direct linkage was successfully established between molecular responses and adverse effects at the organism level, the study has enhanced the understanding of the MoA of single radionuclides and mixtures of these.

Transcriptional changes in Atlantic salmon (Salmo salar) after embryonic exposure to road salt.

Road salt is extensively used as a deicing chemical in road maintenance during winter and has in certain areas of the world led to density stratifications in lakes and ponds, and adversely impacted aquatic organisms in the recipients of the road run-off. Aquatic vertebrates such as fish have been particularly sensitive during fertilisation, as the fertilisation of eggs involves rapid uptake of the surrounding water, reduction in egg swelling and in ovo exposure to high road salt concentrations. The present study aimed to identify the persistent molecular changes occurring in Atlantic salmon (Salmo salar) eggs after 24h exposure to high concentrations (5000 mg/L) of road salt at fertilisation. The global transcriptional changes were monitored by a 60k salmonid microarray at the eyed egg stage (cleavage stage, 255 degree days after fertilisation) and identified a high number of transcripts being differentially regulated. Functional enrichment, pathway and gene-gene interaction analysis identified that the differentially expressed genes (DEGs) were mainly associated with toxiciologically relevant processes involved in osmoregulation, ionregulation, oxidative stress, metabolism (energy turnover), renal function and developmental in the embryos. Quantitative rtPCR analysis of selected biomarkers, identified by global transcriptomics, were monitored in the eggs for an extended range of road salt concentrations (0, 50, 100, 500 and 5000 mg/L) and revealed a positive concentration-dependent increase in cypa14, a gene involved in lipid turnover and renal function, and nav1, a gene involved in neuraxonal development. Biomarkers for osmoregulatory responses such as atp1a2, the gene encoding the main sodium/potassium ATP-fueled transporter for chloride ions, and txdc9, a gene involved in regulation of cell redox homeostasis (oxidative stress), displayed apparent concentration-dependency with exposure, although large variance in the control group precluded robust statistical discrimination between the groups. A No Transcriptional Effect Level (NOTEL) of 50mg/L road salt was found to be several orders of magnitude lower than the adverse effects documented in developing fish embryos elsewhere, albeit at concentrations realistic in lotic systems receiving run-off from road salt. It remains to be determined whether these transcriptional changes may cause adverse effects in fish at ecologically relevant exposure concentrations of road salt.