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Abstract Fish consumption is an important route of exposure to persistent organic pollutants (POPs) in dolphins as well as humans. In order to assess the potential risks associated with these contaminants, 39 whole fish and 37 fillets from fish representing species consumed by dolphins and humans captured from Charleston Harbor and tributaries, South Carolina, USA, were measured for a suite of POPs. Polychlorinated biphenyls (PCBs) were the predominant contaminant with concentrations ranging from 5.02 to 232.20 ng/g in whole fish and 5.42–131.95 ng/g in fillets (weight weight ww) followed by total organochlorine pesticides (OCPs) and polybrominated diphenyl ethers (PBDEs). Total POPs levels varied by location and species with general trends indicating significantly higher levels in fish from the Cooper (93.4 ng/g ww) and Ashley Rivers (56.2 ng/g ww) compared to Charleston Harbor (31.6 ng/g ww). Mullet and spot were found to have significantly higher PCBs, OCPs and total POPs, 2–3 times higher than red drum; mullet were also significantly higher in OCPs compared to seatrout. PCB concentrations in whole fish and fillets exceeded EPA human screening values for cancer risk in all fish sampled. For PCBs in fillets, all samples had values of maximum allowable meals per month that were less than the EPA, FDA guidelines for recommended fish meals per month, suggesting lower (more stringent) allowable fish meals per month. All fish exceeded PBDE wildlife values and all fish except two exceeded the level where 95% of the dolphin population would have tissue levels below the health effect threshold. Considering that POP concentrations in fish potentially consumed by humans exceed human health effect thresholds levels, consumption advisories should be considered as a prudent public health measure. Graphical abstract fx1 Highlights • Higher contaminant levels in fish from Cooper/Ashley Rivers than Charleston Harbor. • Mullet and spot were significantly higher in PCBs and pesticides than red drum. • PCBs were the predominant contaminant in whole fish and fillets from Charleston SC. • PCBs in all fillets exceed EPA human health values, consumption advisories warranted. • Whole fish PCBs, PBDEs exceed wildlife screening values, a concern for local dolphins. [ABSTRACT FROM AUTHOR]

The former lead-zinc mine at Mestersvig, Greenland, continues to contaminate the surrounding environment despite its operations ceasing over 50 years ago. Elevated concentrations of heavy metals are found in water, sediment and biota in the terrestrial, freshwater and marine environments. To shed light on the present contamination and its potential effects on local fish we investigated gill and liver histology of sculpins ( Myoxocephalus spp.) around the former mining area. Two species of sculpins were caught; shorthorn sculpins ( M. scorpius ; n = 16) and fourhorn sculpins ( M. quadricornis ; n = 17) at a contaminated site, Nyhavn, and shorthorn sculpins ( M. scorpius ; n = 25) at the reference site. In a previous study we found histopathological changes in the tissues of the sculpins, and we suspected this to be related to elevated heavy metal tissue concentrations. Concentrations of Fe, Hg, Mn, Pb, Se and Zn were significantly higher in sculpins at Nyhavn compared to the reference site. Reference NOED and LOEC thresholds for biochemistry, tissue lesions, growth, survival and reproduction for hepatic Hg, As, Cd and Pb from the ERED database were exceeded in both sculpin species. Histopathological investigations of the sculpins gills revealed significant increases in the prevalence of hyperplastic epithelium, inflammation, intensity of neutral and total mucus cells and chloride cells along with an increased infection of colonial Peritricha. At the contaminated Nyhavn site, fourhorn sculpins had a significantly higher prevalence of chondroplastic tissue and intensity of neutral, mixed and total mucus cells in the gills compared to the shorthorn sculpins. The data indicate that both sculpin species could be useful indicator species for environmental monitoring of metal pollution in Arctic areas. However, confounding effects of gender and species should be investigated further. Effects on other biomarkers as well as baseline measurements should be included in future environmental monitoring efforts around mining activities in Greenland. [ABSTRACT FROM AUTHOR]

The decline in marine and freshwaters catches in recent years in Colombia has led to a change in dietary habits, with an increase in the purchase and consumption of imported fish. This is of particular concern as fish are sometimes caught in mercury-contaminated waters, and are subsequently sold canned or uncanned. In addition, canned tuna has received little attention as it is widely assumed that concentrations are low. In this study, total mercury (THg) and methylmercury (MeHg) concentrations were evaluated in three imported fish species marketed in Colombia, Prochilodus lineatus, Prochilodus reticulatus, and Pangasianodon hypophthalmus, plus four brands of canned tuna and one of sardines. One brand of tuna showed the highest mean concentrations of THg (0.543 ± 0.237 μg/g, wet weight, ww) and MeHg (0.518 ± 0.337 μg/g ww), while concentrations in P. hypophthalmus were approximately 30 times lower (≈0.02 µg/g ww). The estimated weekly intake (EWI) in children was above the provisional tolerable weekly intake (PTWI) of MeHg established by the Joint FAO/World Health Organization (WHO) Expert Committee on Food Additives (JECFA) in 2007, 1.6 μg/kg body weight (bw) per week, for all the canned tuna brands. Values for adults were below PTWI, whereas for women of childbearing age, values were above PTWI only for brand D of canned tuna. The estimate of the potential risk indicated that MeHg levels in canned tuna can generate negative effects in vulnerable groups, while the EWI of fresh fish did not pose a threat to the general population. Therefore, establishing strategies to address the high consumption of canned tuna, and continuous monitoring to control commercial food, are recommended to decrease Hg exposure. [ABSTRACT FROM AUTHOR]

To tackle the ever-present global concern regarding human exposure to persistent organic pollutants (POPs) via food products, this study strived to indicate associations between organochlorine pesticides (OCPs) and polychlorinated biphenyls (PCBs) in lake-fish tissue depending on the species and sampling season. Apart from the monitoring initiatives recommended in the Global Monitoring Plan for POPs, the study discussed 7 OCPs and 18 PCB congeners determined in three Cyprinidae species (rudd, carp, and Prussian carp) from Vransko Lake (Croatia), which are widely domesticated and reared as food fish across Europe and Asia. We exploit advanced classification algorithms, the Kohonen self-organizing maps (SOM) and Decision Trees (DT), to search for POP patterns typical for the investigated species. As indicated by SOM, some of the dioxin-like and non-dioxin-like PCBs (PCB-28, PCB-74, PCB-52, PCB-101, PCB-105, PCB-114, PCB-118, PCB-156 and PCB-157), α-HCH and β-HCH caused dissimilarities among fish species, but regardless of their weight and length. To support these suggestions, DT analysis sequenced the fish species and seasons based on the concentration of heavier congeners. The presented assumptions indicated that the supplemental application of SOM and DT offers advantageous features over the usually rough interpretation of POPs pattern and over the single use of the methods. [ABSTRACT FROM AUTHOR]

To trace the most concerned bioavailable mercury (Hg) in aquatic environment, fish samples were collected from three typical regions in China, including 3 rivers and 1 lake in the Tibetan Plateau (TP, a high altitude background region with strong solar radiation), the Three Gorges Reservoir (TGR, the largest artificial freshwater reservoir in China), and the Chinese Bohai Sea (CBS, a heavily human-impacted semi-enclosed sea). The Hg isotopic compositions in fish muscles were analyzed. The results showed that anthropogenic emissions were the main sources of Hg in fish from TGR and CBS because of the observed negative δ 202 Hg and positive Δ 199 Hg in these two regions (TGR, δ 202 Hg: − 0.72 to − 0.29‰, Δ 199 Hg: 0.15 – 0.52‰; CBS, δ 202 Hg: − 2.09 to − 0.86‰, Δ 199 Hg: 0.07 – 0.52‰). The relatively higher δ 202 Hg and Δ 199 Hg (δ 202 Hg: − 0.37 – 0.08‰, Δ 199 Hg: 0.50 – 1.89‰) in fish from TP suggested the insignificant disturbance from local anthropogenic activities. The larger slopes of Δ 199 Hg/Δ 201 Hg in fish from TGR (1.29 ± 0.14, 1 SD) and TP (1.25 ± 0.06, 1 SD) indicated methylmercury (MeHg) was produced and photo-reduced in the water column before incorporation into the fish. In contrast, the photoreduction of Hg 2+ was the main process in CBS (slope of Δ 199 Hg/Δ 201 Hg: 1.06 ± 0.06, 1 SD). According to the fingerprint data of Hg isotopes, the most important source for aquatic bioavailable Hg in TP should be the long-range transported Hg, contrasting to the anthropogenic originated MeHg from surface sediments and runoffs in TGR and inorganic Hg from continental inputs in CBS. Therefore, the isotopic signatures of Hg in fish can provide novel clues in tracing sources and behaviors of bioavailable Hg in aquatic systems, which are critical for further understanding the biogeochemical cycling of Hg. [ABSTRACT FROM AUTHOR]

Selenium pollution from the E.W. Brown Electric Generating Station was investigated in Herrington Lake, KY. Coal ash wastewater is discharged as surface water overflow from ash disposal ponds into the lake via a National Pollutant Discharge Elimination System permit issued by the Kentucky Division of Water, but the permit does not restrict or limit the amount of selenium released. Unpermitted discharges occur from seeps and drainage through leaks in ash pond dams. Together, these discharges have resulted in selenium concentrations in water, sediment, benthic macroinvertebrates, and fish that are 2–9 times the level that is toxic for fish reproduction and survival. A large proportion (12.2%, or 25 times background) of juvenile largemouth bass ( Micropterus salmoides , the only species examined) exhibited spinal and/or craniofacial malformations that are consistent with selenium poisoning. Teratogenic Deformity Index values indicated a 3.05% population-level impact on the bass fishery, with total selenium-induced mortality (including pre-swimup mortality) estimated to be in excess of 25% per year. These findings confirm that coal ash discharges into Herrington Lake are contributing selenium to the Lake that is poisoning fish. [ABSTRACT FROM AUTHOR]

Carbon Nanotubes are among the most promising materials for the technology industry. Their unique physical and chemical proprieties may reduce the production costs and improve the efficiency of a large range of products. However, the same characteristics that have made nanomaterials interesting for industry may be responsible for inducing toxic effects on the aquatic organisms. Since the carbon nanotubes toxicity is still a controversial issue, we performed tests of acute and subchronic exposure to a commercial sample of multiwalled carbon nanotubes in two fish species, an exotic model ( Danio rerio ) and a native one ( Astyanax altiparanae ). Using the alkaline version of the comet assay on erythrocytes and the piscine micronucleous, also performed on erythrocytes, it was verified that the tested carbon nanotubes sample did not generate apparent genotoxicity by means of single/double DNA strand break or clastogenic/aneugenic effects over any of the species, independently of the exposure period. Although, our findings indicate the possibility of the occurrence of CNTs-DNA crosslinks. Apparently, the sample tested induces oxidative stress after subchronic exposure as shown by activity of superoxide dismutase and catalase. The data obtained by the activity levels of acetylcholinesterase suggests acute neurotoxicity in Astyanax altiparanae and subchronic neurotoxicity in Danio rerio . [ABSTRACT FROM AUTHOR]

The present study reports total mercury (THg) and methylmercury (MeHg) concentrations in 32 different lean fish species from the Western Mediterranean Sea, with a special focus on the Balearic Islands. The concentrations of THg ranged between 0.05 mg/kg ww and 3.1 mg/kg ww (mean 0.41 mg/kg ww). A considerable number of the most frequently fish species consumed by the Spanish population exceed the maximum levels proposed by the European legislation when they originate from the Mediterranean Sea, such as dusky grouper (100% of the examined specimens), common dentex (65%), conger (45%), common sole (38%), hake (26%) and angler (15%), among others. The estimated weekly intakes (EWI) in children (7–12 years of age) and adults from the Spanish population (2.7 µg/kg bw and 2.1 µg/kg bw, respectively) for population only consuming Mediterranean fish were below the provisional tolerable weekly intake (PTWI) of THg established by EFSA in 2012, 4 µg/kg bw. However, the equivalent estimations for methylmercury, involving PTWI of 1.3 µg/kg bw, were two times higher in children and above 50% in adults. For hake, sole, angler and dusky grouper, the most frequently consumed fish, the estimated weekly intakes in both children and adults were below the maximum levels accepted. These intakes correspond to maximum potential estimations because fish from non-Mediterranean origin is often consumed by the Spanish population including the one from the Balearic Islands. [ABSTRACT FROM AUTHOR]

Exposure to contaminants in fish may be associated with adverse health outcomes even as fish consumption is generally considered beneficial. Risk assessments conducted to support regulatory analyses rely on quantitative fish consumption estimates. Here we report the results of a national survey of high-frequency fish consumers (n = 2099) based on a survey population statistically representative of ~17.6 million U.S. individuals consuming three or more fish meals per week. The survey was conducted during 2013 using an on-line survey instrument. Total fish consumption averaged 111 g/day from market, restaurant and self-caught sources. Depending on the season, the incidence of individuals reporting consumption of self-caught species ranged between 10–12% of our high-frequency fish consuming demographic, averaging approximately 30 g/day and comprising 23% of total fish consumption from all sources of fish. Recreational or self-caught consumption rates vary regionally and are poorly understood, particularly for high-frequency consumers, making it difficult to support national-scale assessments. A divergence between sport-fishing and harvesting of fish as a food-staple is apparent in survey results given differences in consumption patterns with income and education. Highest consumption rates were reported for low income respondents more likely to harvest fish as a food staple. By contrast, the incidence of self-caught fish consumption was higher with income and education although overall consumption rates were lower. Regional differences were evident, with respondents from the East-South Central and New England regions reporting lowest consumption rates from self-caught fish on the order of 12–16 g/day and those from Mountain, Pacific and Mid-Atlantic regions reporting highest rates ranging from 44 to 59 g/day. Respondent-specific consumption rates together with national-level data on fish tissue concentrations of PCBs, MeHg, and PFOS suggest that 10–58% of respondents reporting self-caught fish consumption are exposed to concentrations of these contaminants that exceed threshold levels for health effects based on a target hazard index of one, representing 2.3 M to 19 M individuals. The results of this nationwide survey of high-frequency fish consumers highlights regional and demographic differences in self-caught and total fish consumption useful for policy analysis with implications for distributional differences in potential health impacts in the context of both contaminant exposures as well as protective effects. [ABSTRACT FROM AUTHOR]

The aim of this study was to investigate and compare total mercury (Hg), selenium (Se), and Se:Hg molar ratios in fish muscles (phytophages n=3; benthophages n=32; predators n=5) and semiaquatic carnivores, including piscivores (the European otter n=8, the feral American mink n=7) and the omnivorous raccoon (n=37) from a riverine European ecosystem in a Se-deficient area. The Hg concentration in fish reached 0.337 μg/g dry weight, dw (0.084 μg/g wet weight, ww). We found significant differences among Hg levels in tested vertebrate groups (predators vs benthophages: 0.893 vs 0.281 μg/g; piscivores vs omnivores: 6.085 vs 0.566 μg/g dw). Fish groups did not differ in Se concentrations, with a mean value of 0.653 μg/g dw. Significant differences were revealed between Se levels in piscivorous and omnivorous carnivores (0.360 vs 0.786 μg/g dw, respectively). Fish Se:Hg molar ratio values were >2.2. Benthophages had higher the ratio than predators but similar to phytophages. Among carnivores, piscivores had much lower the ratio than raccoon (0.14 vs 3.75) but raccoon and fish medians did not significantly differ. We found almost two times higher Se levels in fish and raccoons compared to piscivores, possibly resulting from lower fish Se digestibility by piscivores in contrast to higher absorption of plant Se by many fish and omnivorous raccoons. Considering that a tissue Se:Hg molar ratio <1 may be connected with a Hg toxicity potential increase, we assume that piscivores in Se-deficient area are in worse situation and more exposed to Hg than fish and omnivores. [ABSTRACT FROM AUTHOR]

Metal bioaccumulation and induction of biomarkers such as lipid peroxidation (LPO), superoxide dismutase (SOD), catalase (CAT), glutathione S transferase (GST), reduced glutathione (GSH) and DNA damage are potential indicators of stress in Channa punctatus exposed to effluents. In canal water, receiving thermal power plant discharges, Fe and Ni concentrations exceeded the recommended guidelines set by the United Nations Environment Programme Global Environment Monitoring System (UNEPGEMS). Fe was highly bioavailable and accumulated in all organs (liver, kidney, muscle and integument). The highest metal pollution index (MPI) value of 41.2 was observed in kidney and the lowest 13.5 in muscle tissue. LPO, SOD, CAT and GST levels were significantly higher in liver and kidney, whereas GSH levels declined significantly compared to fish from the reference site. Concomitant damage to DNA was observed with significantly higher mean tail length in the exposed fish gill cells (26.5 µm) and in liver (20.8 µm) compared to reference fish. Therefore, it can be concluded that the thermal power plant effluent had the potential to cause oxidative stress and DNA damage in C. punctatus . [ABSTRACT FROM AUTHOR]

The extensive use of malathion, an organophosphate pesticide, raises the possibility of its undesirable toxicity to non-target organisms. Agricultural run-off and vector control sprays are the major sources of exposure to this pesticide for aquatic organisms. Some earlier studies have reported the presence of malathion at concentrations ranging from 18.12 μg/L to 105.2 μg/L in various water samples. In this study, we have tested the hypothesis that these sub-lethal yet environmentally significant concentrations of malathion has serious toxicological implications on the fingerlings of Labeo rohita. Exposure to increasing concentration of malathion (10, 50 and 100 μg/L) was reflected in the serum concentration of the pesticide and also in the inhibition of acetylcholinesterase activity in fish brains. Increased abnormalities in liver function test coupled with a rise in the oxidative stress response were observed in gills, liver and kidney. However, the increase in antioxidant enzyme activities like superoxide dismutase and glutathione-S-transferase by malathion exposure suggested a hormetic response. Tissue injury due to malathion was evident from the morphological and nuclear anomalies in the H-E stained sections of gill, liver and kidney. Cell cycle analysis of these organs further fortified the histopathological findings. This study elucidates the sub-lethal toxicity of environmentally relevant malathion concentrations on Labeo rohita which indicates the potential health hazard posed to human beings consuming this fish. This calls for careful application of malathion in areas adjoining to inland fisheries. [ABSTRACT FROM AUTHOR]

The crown-of-thorns starfish Acanthaster planci is a venomous starfish whose venom provokes strong cytotoxicity. In the present study, the purified cytotoxic toxin of A. planci venom (CAV) was identified as plancitoxin I protein by mass spectrum analyses. This study aims to investigate the molecular mechanism underlying the cytotoxicity function of plancitoxin I by focusing on the oxidative stress, mitochondrial dysfunction and endoplasmic reticulum (ER) stress pathway in human melanoma A375.S2 cells. The results indicated that after being treated with CAV toxin, A375.S2 cells significantly decreased viability in a dose-dependent manner. The CAV was found to reduce the cellular antioxidant enzymes such as SOD and CAT, and there was a significant decrease in total thiol level and mtDNA integrity, and it enhanced the lipid peroxidation. In addition, CAV increased cytosolic Ca 2 + concentration, and enhanced the expression of the ER molecular chaperones GRP78 and CHOP in a dose-dependent manner. CAV significantly elevated the activity of caspase-3, -8 and -9, and reduced the ratio of Bcl-2/Bax. The cells exhibited apoptosis were determined by using propidium iodide (PI) staining of DNA fragmentation (sub-G1 peak). In summary, the results demonstrated that plancitoxin I inhibits the proliferation of A375.S2 cells through induction of oxidative stress, mitochondrial dysfunction and ER stress associated apoptosis. [ABSTRACT FROM AUTHOR]

The aim of this paper was to investigate baseline toxicity to rats and effect of exposure routes on toxicity in rats and fish. In this paper, 1588 industrial chemicals were selected to investigate baseline toxicity to rats. The results showed that rat toxicity varies around a constant for classified compounds or homologues. The toxic contributions of substituted functional groups have been calculated and alkanes were used as baseline toxicity. The toxic contributions, equal to toxic ratios (TR), show that small changes in chemical structure can result in different toxic effect in rat toxicity. However, this situation has not been observed in fish toxicity because the threshold of excess toxicity (e.g. log TR = 1) was too high to distinguish differences in toxicity. Very close critical body residues (CBRs) calculated from percentage of absorption and bioconcentration factors indicate that most of aliphatic chemicals may share the same modes of toxic action between rat and fish species. The high estimation error of bioconcentration factor calculated from computer programs for some compounds suggests that classification of excess toxicity should be based on the CBRs, rather than the TR because the TR is closely related to the exposure routes. [ABSTRACT FROM AUTHOR]

Distribution and marketing of chemicals require appropriate labelling of health, physical and environmental hazards according to the United Nations global harmonisation system (GHS). Labelling for (human) acute toxicity categories is based on experimental findings usually obtained by oral, dermal or inhalative exposure of rodents. There is a strong societal demand for replacing animal experiments conducted for safety assessment of chemicals. Fish embryos are considered as alternative to animal testing and are proposed as predictive model both for environmental and human health effects. Therefore, we tested whether LC50s of the fish embryo acute toxicity test would allow effectively predicting of acute mammalian toxicity categories. A database of published fish embryo LC50 containing 641 compounds was established. For these compounds corresponding rat oral LD50 were identified resulting in 364 compounds for which both fish embryo LC50 and rat LD50 was available. Only a weak correlation of fish embryo LC50 and rat oral LD50 was obtained. Fish embryos were also not able to effectively predict GHS oral acute toxicity categories. We concluded that due to fundamental exposure protocol differences (single oral dose versus water-borne exposure) a reverse dosimetry approach is needed to explore the predictive capacity of fish embryos. [ABSTRACT FROM AUTHOR]

Highlights: [•] Alternatives are yet only available for acute fish toxicity testing. [•] Limited access to animal test data hinders the development of alternatives. [•] Successful validation of alternative approaches is required to replace animal tests. [•] Mechanistic approaches should be developed and implemented in regulatory testing. [•] Integrated testing strategies combine experimental and non-testing alternatives. [Copyright &y& Elsevier]

Abstract: Goliath grouper have undergone significant global population declines with potential biological extinction for some subpopulations. Although overfishing and habitat loss are important drivers of these declines, the negative effects of contaminants may also play a role. The life history patterns of goliath grouper may make this species especially prone to exposure to contaminants and may exacerbate bioaccumulation of toxic substances, including mercury, which has documented detrimental health effects. Therefore, we analyzed mercury (in muscle, liver, kidney, gonad, and brain tissue) and the histology of key organs (liver, kidney and gill tissue) in 56 goliath groupers from U.S. waters. Total mercury concentration was greatest in liver tissue, followed by kidney, muscle, gonad, and brain. Maximum mercury concentration ranged from 22.68μg/g in liver tissue to 0.89μg/g in brain tissue. Mean mercury concentration ranged from 2.87μg/g in liver tissue to 0.37μg/g in brain tissue with a mean of 0.63μg/g in muscle. Mean mercury concentrations observed in goliath grouper from U.S. waters were within the range known to cause direct health effects in fish after long-term exposure. The lesions and histological changes observed in the liver, kidney, and gills of goliath groupers were similar to those found in other fish following laboratory mercury-exposure trials and to those found in mercury-contaminated fish in wild populations carrying similar or even lower concentrations. We suggest that exposure to mercury and other environmental influences such as pathogens and reduced temperatures could be co-factors in the histological effects or anomalies observed in the present study, and resulting stresses may be involved in the observed population declines in the species. [Copyright &y& Elsevier]

Abstract: Squirrelfishes and soldierfishes (Holocentridae) are among the most conspicuous species in the nocturnal reef fish community. However, there is no clear consensus regarding their evolutionary relationships, which is reflected in a complicated taxonomic history. We collected DNA sequence data from multiple single copy nuclear genes and one mitochondrial gene sampled from over fifty percent of the recognized holocentrid species and infer the first species-level phylogeny of the Holocentridae. Our results strongly support the monophyly of the clades Myripristinae (soldierfishes) and Holocentrinae (squirrelfishes). The molecular phylogenies differ with regard to previous hypotheses of relationships within the Myriprisitinae, resolving a clade of cryptic reef associated and deep water non-reef dwelling lineages (Corniger + Plectrypops + Ostichthys) that is the sister lineage to a monophyletic Myripristis. Within Holocentrinae, Neoniphon and Sargocentron are strongly supported as paraphyletic, while Holocentrus is nested within Sargocentron. Using Bayesian ancestral state reconstruction methods, we demonstrate the taxonomically diagnostic characters for Neoniphon and Sargocentron likely represent character states with a complex evolutionary history that is not reflective of shared common ancestry. We propose a new classification for Holocentrinae, recognizing four lineages that are treated as genera: SargocentronFowler, 1904, HolocentrusScopoli, 1777, FlameoJordan and Evermann, 1898, and NeoniphonCastelnau, 1875. [Copyright &y& Elsevier]

Abstract: Caspase-8 is an initiator caspase that plays a crucial role in some cases of apoptosis by extrinsic and intrinsic pathways. Caspase-8 structure and function have been extensively studied in mammals, but in fish the characterization of that initiator caspase is still scarce. In this work, the sea bass counterpart of mammalian caspase-8 was sequenced and characterized, and its involvement in the apoptogenic activity of a toxin from a fish pathogen was assessed. A 2472 bp cDNA of sea bass caspase-8 was obtained, consisting of 1455 bp open reading frame coding for 484 amino acids and with a predicted molecular weight of 55.2 kDa. The sea bass caspase-8 gene has 6639 bp and is organized in 11 introns and 12 exons. Several distinctive features of sea bass caspase-8 were identified, which include two death effector domains, the caspase family domains p20 and p10, the caspase-8 active-site pentapeptide and potential aspartic acid cleavage sites. The sea bass caspase-8 sequence revealed a significant degree of similarity to corresponding sequences from several vertebrate taxonomic groups. A low expression of sea bass caspase-8 was detected in various tissues of non-stimulated sea bass. Furthermore, it is shown that stimulation of sea bass with mid-exponential phase culture supernatants from Photobacterium damselae ssp. piscicida (Phdp), known to induce selective apoptosis of macrophages and neutrophils, resulted in an increased expression of caspase-8 in the spleen, one of the main affected organs by Phdp infection. [Copyright &y& Elsevier]

Fish toxicity testing has been conducted since the 1860's in order to help define safe levels of chemical contaminants in lakes, rivers and coastal waters. The historical emphasis on acute lethality testing of chemicals has more recently focussed on long term sublethal effects of chemicals on fish and their prey species. Fish toxicity testing is now embedded in much environment legislation on chemical safety while it is recognized that animal use should be Replaced, Reduced and Refined (the 3Rs) where possible. The OECD Fish Toxicity Testing Framework provides a useful structure with which to address the needs of environmental safety assessment whilst implementing the 3Rs. This commentary aims to promote the implementation of the recommendations of the OECD Fish Toxicity Testing Framework. [ABSTRACT FROM AUTHOR]

Abstract: The variability of toxicity data contained within databases was investigated using the widely used US EPA ECOTOX database as an example. Fish acute lethality (LC50) values for 44 compounds (for which at least 10 data entries existed) were extracted from the ECOTOX database yielding a total of 4654 test records. Significant variability of LC50 test results was observed, exceeding several orders of magnitude. In an attempt to systematically explore potential causes of the data variability, the influence of biological factors (such as test species or life stages) and physical factors (such as water temperature, pH or water hardness) were examined. Even after eliminating the influence of these inherent factors, considerable data variability remained, suggesting an important role of factors relating to technical and measurement procedures. The analysis, however, was limited by pronounced gaps in the test documentation. Of the 4654 extracted test reports, 66.5% provided no information on the fish life stage used for testing. Likewise, water temperature, hardness or pH were not recorded in 19.6%, 48.2% and 41.2% of the data entries, respectively. From these findings, we recommend the rigorous control of data entries ensuring complete recording of testing conditions. A more consistent database will help to better discriminate between technical and natural variability of the test data, which is of importance in ecological risk assessment for extrapolation from laboratory tests to the field, and also might help to develop correction factors that account for systematic differences in test results caused by species, life stage or test conditions. [Copyright &y& Elsevier]

Abstract: To investigate the possible involvement of the nitric oxide radical (NO) in ciguatera fish poisoning (CFP), the in vitro effects of the main Pacific ciguatoxin (P-CTX-1B) and bacterial lipopolysaccharide (LPS) were comparatively studied on neuroblastoma Neuro-2a and on macrophage RAW 264.7 cell lines. NO accumulation was quantified by measuring nitrite levels in cellular supernatant using Griess reagent while the up-regulation of inducible nitric oxide synthase (iNOS) at the mRNA level was quantified via Real-Time Reverse-Transcription Polymerase Chain Reaction (RT-PCR). P-CTX-1B caused a concentration- and time-dependent induction of iNOS in RAW 264.7 cells but not in Neuro-2a cells. NO production was evidenced by increased nitrite levels in the 10μM range after 48h of RAW 264.7 cells exposure to LPS and P-CTX-1B (0.05μg/ml and 6nM, respectively). The expression of iNOS mRNA peaked at 8h for LPS then gradually decreased to low level at 48h. In contrast, a sustained level was recorded with P-CTX-1B in the 8–48h time interval. The addition of N ω-nitro-l-arginine methyl ester (l-NAME), a stereoselective NOS inhibitor, strongly diminished NO formation but had no effect on iNOS mRNA synthesis. The implication of NO in CFP paves the way for new therapies for both western and traditional medicines. [Copyright &y& Elsevier]