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).

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.

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.

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.

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.

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 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.

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.

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.

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.

Global transcriptional analysis of short-term hepatic stress responses in Atlantic salmon (Salmo salar) exposed to depleted uranium.

Potential environmental hazards of radionuclides are often studied at the individual level. Sufficient toxicogenomics data at the molecular/cellular level for understanding the effects and modes of toxic action (MoAs) of radionuclide is still lacking. The current article introduces transcriptomic data generated from a recent ecotoxicological study, with the aims to characterize the MoAs of a metallic radionuclide, deplete uranium (DU) in an ecologically and commercially important fish species, Atlantic salmon (Salmo salar). Salmon were exposed to three concentrations (0.25, 0.5 and 1.0 mg/L) of DU for 48 h. Short-term global transcriptional responses were studied using Agilent custom-designed high density 60,000-feature (60 k) salmonid oligonucleotide microarrays (oligoarray). The microarray datasets deposited at Gene Expression Omnibus (GEO ID: GSE58824) were associated with a recently published study by Song et al. (2014) in BMC Genomics. The authors describe the experimental data herein to build a platform for better understanding the toxic mechanisms and ecological hazard of radionuclides such as DU in fish.

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.

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).

Sublethal effects in Atlantic salmon (Salmo salar) exposed to mixtures of copper, aluminium and gamma radiation.

The present study was designed to investigate the effects in presmolt of Atlantic salmon (Salmo salar) exposed to copper (Cu), aluminium (Al) and gamma radiation, individually or in combination. Fish were exposed for 48 h to metals added to lake water; 10, 40 and 80 µg Cu/L, 250 µg Al/L and a combination of 40 µg Cu/L and 250 µg Al/L. In addition, gamma radiation (4-70 mGy delivered over 48 h) was added as an additional exposure stressor. Selected endpoints were chosen to reveal different toxic mechanisms and included Cu and Al accumulation on gills, blood chemistry and haematological variables (plasma sodium and chloride, haematocrit, glucose), hepatic levels of reduced and oxidised glutathione (GSH and GSSG) and hepatic transcriptional response of glutathione peroxidase (GPx), gamma-glutamylcysteine synthetase (GCS), metallothionein (MT) and ubiquitin. Exposure to Cu alone resulted in gill accumulation of Cu, reduction of plasma ions and increased transcriptional response of GPx, MT and ubiquitin. Exposure to Al alone reduced plasma ion levels but did not affect any of the hepatic biomarkers except for ubiquitin. The combined metal exposure (Cu + Al) altered the GSH levels, however GPx and MT were not affected suggesting a different mode of detoxification in the combined exposure. Gamma radiation appeared to influence GSH and ubiquitin levels. The observed effects seemed to be both stressor and concentration dependent.

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.

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.

Bioaccumulation of persistent organic pollutants (POPs) in fish species from Lake Koka, Ethiopia: The influence of lipid content and trophic position.

The concentrations and bioaccumulation of persistent organic pollutants (POPs) were determined in four fish species from Lake Koka, Ethiopia, representing 2-3 levels in the food chain of the lake. Dichlorodiphenyltrichloroethanes (DDTs), endosulfans, polychlorinated biphenyls (PCBs) and chlorpyrifos were identified, with DDTs as the most predominant pesticide, with concentration ranging from 0.05 to 72.53ngg(-1) wet weight (ww). All fish tissue samples collected from different species of the lake contained residues of DDTs. The maximum level of DDTs was found in the fattiest, African sharptooth catfish (Clarias gariepinus) sampled from the lake, with a mean concentration of 15.15ngg(-1)ww. The significant (P<0.05) relationship between concentrations of DDTs and δ(15)N indicates that DDTs biomagnified in the food web of the lake. The 4,4'-DDE to 4,4'-DDT ratio in Oreochromis niloticus (0.6) and Cyprinus carpio (0.5) were below 1, indicating ongoing use of DDTs in the study area and recent exposure of these fish species.

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.