Aquatic Toxicology

The influence of long-term exposure of goldfish to dietary cadmium (Cd) on its accumulation in tissues, growth, ovarian development, luteinizing hormone (LH) secretion and a response to hormonal stimulation of spawning were evaluated. The study was conducted on four groups of females for the period of 3 years, from the age of 10 weeks to second spawning. Four doses of Cd were applied in the feed: 0 (control group), 0.1, 1 and 10 mg Cd g −1 of feed (wet weight). The highest dose of Cd (10 mg g −1) inhibited growth and caused several behavioural effects. In contrast, lower dose of Cd (1 mg g −1) stimulated fish growth. The doses of Cd from 0.1 to 1 mg Cd g −1 did not influence ovarian development. The gonado-somatic index (GSI) and histological analysis of ovaries showed no differences in ovarian development between the control group and the groups receiving these doses of Cd. However, in the group receiving the highest Cd dose, GSI decreased. This was associated with persistent, long-lasting elevation of plasma LH levels. Ovulation did not occur in this group. Injections of salmon GnRH-analogue (sGnRHa) alone or with domperidone (a dopamine receptor antagonist) in sexually mature fish caused an increase of LH levels in all groups, although in the group fed with the highest Cd dose the effect was weaker than in the other groups. After the first spawning season, a negative effect of lower Cd doses (0.1 and 1 mg Cd g −1) on ovarian recrudescence (rebuilding of ovaries) and on the response to the consecutive hormonal stimulation of spawning was observed (lower number of ovulating females). There was a significantly higher content of Cd in the livers of fish than in their muscles. The results of hormonal stimulation of spawning and histological analysis of ovaries suggest that in goldfish cadmium acts mainly at the level of ovary rather than on the pituitary gland. We suppose that in the natural environment cadmium present in the feed can play an important role in the accumulation of this element in fish tissues and can influence vital physiological processes.

In ecotoxicological studies involving community-level investigations, rapid and multiparametric fluorescence-based methods may provide substantial advantages over traditional methods used for structural and functional community analysis. Therefore, multiwavelength-excitation pulse-amplitude modulated (PAM) fluorometry was applied in this study to assess long-term changes in periphyton community structure, shortterm effects on periphyton functioning (photosynthesis) and pollution induced community tolerance (PICT). For inter-calibration, periphyton structure was evaluated by chemotaxonomic analysis of accessory pigments and a four-wavelength-excitation PAM fluorometer. Short-term effects of herbicides were evaluated by fluorescence quenching analysis and 14 C-incorporation as a proxy of primary production. Subsequently, the method was applied to assess structural and functional changes in periphyton communities after isoproturon exposure for 14 and 26 days, respectively. Results showed good correlation of the PAM fluorescence-based measurements with traditional methods for biofilms in the initial colonisation phase for structural and functional parameters. However, for biofilms older than 9 weeks PAM fluorescence may underestimate biomass. Multiwavelength-excitation PAM fluorometry showed good correlation with marker pigment concentrations indicating that this method provides a reliable estimate of the community structure. PAM fluorometry was able to quantify changes of biomass and follow relative shifts in class composition of biofilms under exposure of isoproturon. Short-term tests based on the quantification of the inhibition of the effective quantum yield revealed a concentration-dependent increase of PICT. The observation of two succession phases of the biofilms after 14 and 26 days of growth, respectively, revealed that sensitivity of biofilms decreased with increasing age and biomass, respectively, but PICT remained a characteristic parameter of exposed communities in a concentration-dependent relationship. In conclusion, multiwavelength-excitation PAM fluorometry has considerable potentials in multispecies and multiparameter assessment of toxic effects on community level in terms of (1) a combined and rapid evaluation of structural and functional parameters in parallel, (2) screening of trends over time, (3) observing effects in replication and (4) being non-destructive. The approach therefore provides a perspective for a better understanding of community-level effects as species interactions in terms of PICT and therefore a higher ecological realism in risk assessment of toxicants.

Recently, a novel approach to a highly sensitive and quantitative detection of rare earth element (REE) ions including La3+, Eu3+ and Tb3+, by the polymerase chain reaction (PCR) technique, has been reported. The detection of REE ions is based on the catalytic nature of REE ions targeting the deoxyribonucleic acid (DNA), thus monitoring of the ions can be achieved by reading the level of intact DNA by PCR. Despite of its high sensitivity (at ppb to ppt levels), the conventional PCR-based REE detection protocol requires certain length of time (1-2 hours). In the present study, we modified the PCR-based REE detection protocols by employing the high-speed PCR, and performed the automated and rapid detection of La3+ in small-sized aqueous samples within 5min.

50-day's free access https://authors.elsevier.com/a/1dnnLB8ccr0ST ........................................................................................................................................... Cyanobacteria are notorious for producing harmful algal blooms that present an ever-increasing serious threat to aquatic ecosystems worldwide, impacting the quality of drinking water and disrupting the recreational use of many water bodies. Remote sensing techniques for the detection and quantification of cyanobacterial blooms are required to monitor their initiation and spatiotemporal variability. In this study, we developed a novel semi-analytical approach to estimate the concentration of cyanobacteria-specific pigment phycocyanin (PC) and common phytoplankton pigment chlorophyll a (Chl a) from hyperspectral remote sensing data. The PC algorithm was derived from absorbance-concentration relationship, and the Chl a algorithm was devised based on a conceptual three-band structure model. The developed algorithms were applied to satellite imageries obtained by the Hyperspectral Imager for the Coastal Ocean (HICO™) sensor and tested in Lake Kinneret (Israel) during strong cyanobacterium Microcystis sp. bloom and out-of-bloom times. The sensitivity of the algorithms to errors was evaluated. The Chl a and PC concentrations were estimated with a mean absolute percentage difference (MAPD) of 16% and 28%, respectively. Sensitivity analysis shows that the influences of backscattering and other water constituents do not affect the estimation accuracy of PC (~2% MAPD). The reliable PC/Chl a ratios can be obtained at PC concentrations above 10 mg m−3. The computed PC/Chl a ratio depicts the contribution of cyanobacteria to the total phytoplankton biomass and permits investigating the role of ambient factors in the formation of a complex planktonic community. The novel algorithms have extensive practical applicability and should be suitable for the quantification of PC and Chl a in aquatic ecosystems using hyperspectral remote sensing data as well as data from future multispectral remote sensing satellites, if the respective bands are featured in the sensor.

Fish are particularly sensitive to a wide variety of agrochemicals including glyphosate herbicide that may arise from not only deliberate discharge of these chemicals into waterways but also from approved agricultural practices. In the present study, the toxicity of glyphosate an herbicide was investigated with emphasis on histopathological effects. Nile tilapias (Oreochromis niloticus) juvenile 15 ± 1.0 g were exposed to 0, 2, 9, 30, 97, 310 mg/l of glyphosate. The lethal concentration (LC 50) value of glyphosate was 1.05 mg/l for 96 h of exposure. Glyphosate concentration corresponding to the 96 h LC 50 values for juvenile tilapia was used to study the effects of glyphosate exposure for 4 days in inducing histopathological changes of gills, livers, kidneys and brains. In the gills, filament cell proliferation, lamellar fusion, lamellar cell hyperplasia, and epithelial lifting were observed. In the liver, there was vacuolation of hepatocytes and necrosis. Kidney lesion consisted of hyaline droplets in the tubular epithelial cells and pkynosis. The brain lesion also consisted of generalized neuronal degeneration and spongiosis. The changes in these tissues occur predominantly in the 96 h exposure. Respiratory stress, erratic swimming and instant death of fish were observed in exposed fish, which varied with the concentration of the toxicant and this showed that mortality increased with increase in concentration. Glyphosate is highly toxic to Nile tilapias, which are more susceptible to this herbicide; therefore their use near fish farm or in area close to aquatic environment should be discouraged.

Numerous studies have indicated that aquatic pollution affects parasite populations and communities. However, the responses of individual taxa and functional groups to specific contaminants, and their effect sizes, have yet to be assessed quantitatively. We began by summarizing general trends in the literature, and found that reports of significant changes in parasitism were most commonly observed in response to eutrophication and metal contamination. Among parasite taxa, significant changes were most commonly reported for acanthocephalans, digeneans and microparasites. We then conducted a quantitative metaanalysis of the effects of pollution on parasitism in aquatic animals. We calculated signed standardized effect sizes (as Cohen's d) for all published studies that provided the necessary descriptive statistics, and compared them among major contaminant types (pesticides, hydrocarbons, polychlorinated biphenyls, pulp mill effluent, metals, sewage, eutrophication, and acidification) and parasite taxa (Acanthocephala, Cestoda, Digenea, Monogenea, Nematoda and microparasites). Effect sizes were not significantly different from zero for many parasite/contaminant interactions, and tended to be highly variable within individual taxa. However, consistently strong, significant negative effects were noted in Digenea and Monogenea, especially in response to metal pollution. Significant effect sizes were typically negative, indicating that pollutants have negative effects on parasite populations. Contaminants also had a slightly negative effect on community richness. When parasites were grouped into heteroxenous (with >1 obligatory host in life cycle) vs. monoxenous (1 obligatory host in life cycle) taxa, the latter were more susceptible to a wide range of pollutants. Similarly, directly exposed (external parasites and the free-living transmission stages of internal parasites) and freshwater taxa were more susceptible to a wider range of pollutants than indirectly exposed (internal parasites) and marine taxa. This meta-analysis represented a first attempt to consider the size of the effect of pollution on parasites, and highlighted the potential of susceptible parasite taxa, communities, and functional groups for use in the biological assessment of pollution. For instance, our results indicate that freshwater monogeneans and digeneans are good candidates as potential biological indicators of pollution in aquatic ecosystems.

In order to mimic the biological effects of an oil spill in Arctic waters, we examined several types of biomarkers (genes, enzymes, metabolites, and DNA damage) in polar cod Boreogadus saida experimentally exposed to the water soluble fractions of crude oil. During 4 weeks of exposure, induction of the studied biomarkers exceeded baseline levels. The mRNA expression of the cytochrome P4501A1 (cyp1a1) gene was the most promising biomarker, with glutathione S-transferase (gst) as a suitable complement. The delayed ethoxyresorufin O-deethylase (EROD) and GST activities and their persistence following 2 weeks of depuration may allow detection of previous exposures in field samples. The composition of PAH metabolites in the bile indicated the bioavailability of different PAH size-classes. Although mRNA expressions of antioxidant defense genes were induced at start of the exposure, with the strongest responses from catalase and cytosolic superoxide dismutase, they were poor for oil monitoring purposes due to their very short response times. Significant DNA damage demonstrated genotoxicity even at low PAH concentrations (<15 g L −1 ) and was correlated with benzo(a)pyrene and pyrene metabolites in the bile.

Nuclear radioisotope accidents are potentially ecologically devastating due to their impact on marine organisms. To examine the effects of exposure of a marine organism to radioisotopes, we irradiated the intertidal copepod Tigriopus japonicus with several doses of gamma radiation and analyzed the effects on mortality, fecundity, and molting by assessing antioxidant enzyme activities and gene expression patterns. No mortality was observed at 96h, even in response to exposure to a high dose (800Gy) of radiation, but mortality rate was significantly increased 120h (5 days) after exposure to 600 or 800Gy gamma ray radiation. We observed a dose-dependent reduction in fecundity of ovigerous females; even the group irradiated with 50Gy showed a significant reduction in fecundity, suggesting that gamma rays are likely to have a population level effect. In addition, we observed growth retardation, particularly at the nauplius stage, in individuals after gamma irradiation. In fact, nauplii i...

Environmental pollutants are hypothesized to be one of the causes of recent declines in wild populations of Atlantic salmon (Salmo salar ) across Eastern Canada and the United States. Some of these pollutants, such as polychlorinated biphenyls and dioxins, are known to induce expression of the CYP1A subfamily of genes. We applied a highly sensitive technique, quantitative reverse transcription-polymerase chain reaction (RT-PCR), for measuring the levels of CYP1A induction in Atlantic salmon. This assay was used to detect patterns of CYP1A mRNA levels, a direct measure of CYP1A expression, in Atlantic salmon exposed to pollutants under both laboratory and field conditions. Two groups of salmon were acclimated to 11 and 17 8C, respectively. Each subject then received an intraperitoneal injection (50 mg kg (1 ) of either b-naphthoflavone (BNF) in corn oil (10 mg BNF ml (1 corn oil) or corn oil alone. After 48 h, salmon gill, kidney, liver, and brain were collected for RNA isolation and analysis. All tissues showed induction of CYP1A by BNF. The highest base level of CYP1A expression (2.56)/10 10 molecules/mg RNA) was found in gill tissue. Kidney had the highest mean induction at five orders of magnitude while gill tissue showed the lowest mean induction at two orders of magnitude. The quantitative RT-PCR was also applied to salmon sampled from two streams in Massachusetts, USA. Salmon liver and gill tissue sampled from Millers River (South Royalston, Worcester County), known to contain polychlorinated biphenyls (PCBs), showed on average a two orders of magnitude induction over those collected from a stream with no known contamination (Fourmile Brook, Northfield, Franklin County). Overall, the data show CYP1A exists and is inducible in Atlantic salmon gill, brain, kidney, and liver tissue. In addition, the results obtained demonstrate that quantitative PCR analysis of CYP1A expression is useful in studying ecotoxicity in populations of Atlantic salmon in the wild. #

The platinum group metals (PGMs) platinum (Pt), palladium (Pd), and rhodium (Rh) are used in automobile catalytic converters, from which they have been emitted into the environment to an increasing degree during the last 20 years. Despite the bioavailability of these metals to plants and animals, studies determining the effects of PGMs on organisms are extremely rare. In the present study, effects of various concentrations of PtCl 2 (0.1, 1, 10, 50 and 100 g/L) were investigated with respect to the induction of hsp70 and histopathological alterations in the zebrafish, Danio rerio and the ramshorn snail, Marisa cornuarietis. Histopathological investigations revealed effects of Pt on both species, which varied between slight and strong cellular reactions, depending on the PtCl 2 concentration. The hsp70 level in M. cornuarietis did not show an increase following Pt exposure whereas it was significantly elevated at 100 g/L PtCl 2 in D. rerio.

Dieldrin and p,p′-DDE are ubiquitous contaminants known to act as endocrine disruptors, causing impaired development and reproduction in fish and wildlife. In order to elucidate the mechanisms by which dieldrin and p,p′-DDE cause endocrine disruption in largemouth bass (Micropterus salmoides), fish were exposed subchronically through the diet to both contaminants. Following 120 days of exposure, p,p′-DDE decreased estradiol in females, but increased 11-ketotestosterone in both sexes. Dieldrin on the other hand, decreased estradiol and 11-ketotestosterone in both sexes. Both pesticides also altered steady state mRNA expression levels of a set of genes chosen to represent three possible mechanisms of endocrine disruption: (1) direct interaction with soluble sex steroid receptors, (2) biosynthesis of endogenous sex hormones, and (3) metabolism of endogenous hormones. p,p′-DDE acted as a weak estrogen, increasing the expression of vitellogenin and estrogen receptor α in the liver. p,p′-DDE also altered the expression of genes involved in the synthesis of endogenous hormones as well as their metabolism. Dieldrin, on the other hand, only altered expression of vitellogenin and not estrogen receptor α . Dieldrin also altered the expression of genes involved in hormone synthesis and metabolism, and it dramatically lowered plasma hormone levels. Both pesticides targeted expression of genes involved in all three modes of action, suggesting that they each have multiple modes of action.

Increasing emissions of anthropogenic mercury represents a growing concern to the health of high trophic level marine mammals. In its organic form, this metal bioaccumulates, and can be toxic to several physiological endpoints, including the immune system. In this study, we (1) evaluated the effects of inorganic mercury (mercuric chloride, HgCl 2 ) and organic mercury (methylmercuric chloride, MeHgCl) on the in vitro function of lymphocytes isolated from the peripheral blood of beluga whales (Delphinapterus leucas); (2) characterized the potential protective effects of sodium selenite (Na 2 SeO 3 ) on cell proliferation of HgCl 2 or MeHgCl-treated beluga whale lymphocytes; and (3) compared these dose-dependent effects to measurements of blood Hg in samples collected from traditionally harvested beluga whales in the western Canadian Arctic. Lymphocyte proliferative responses were reduced following exposure to 1 M of HgCl 2 and 0.33 M of MeHgCl. Decreased intracellular thiol levels were observed at 10 M of HgCl 2 and 0.33 M of MeHgCl. Metallothionein induction was noted at 0.33 M of MeHgCl. Concurrent exposure of Se provided a degree of protection against the highest concentrations of inorganic Hg (3.33 and 10 M) or organic Hg (10 M) for T-lymphocytes. This in vitro protection of Se against Hg toxicity to lymphocytes may contribute to the in vivo protection in beluga whales exposed to high Hg concentrations. Current Hg levels in free-ranging beluga whales from the Arctic fall into the range of exposures which elicited effects on lymphocytes in our study, highlighting the potential for effects on host resistance to disease. The implications of a changing Arctic climate on Hg fate in beluga food webs and the consequences for the health of beluga whales remain pressing research needs.

After the definitive ban on tin-based antifouling substances, new organic compounds have recently been introduced in antifouling paint formulations, as either principal or booster biocides. In most cases, previous risk assessment of these biocides has been inadequate so that their possible effects on aquatic ecosystems is a matter of great concern. We studied the effects of two new organic biocides often associated in paint formulations, Sea-Nine 211 TM (4,5 dichloro-2-n-octyl-4-isothiazoline-3-one) and chlorothalonil (2,4,5,6-tetrachloroisophthalonitrile), on haemocytes of the compound ascidian Botryllus schlosseri exposed for 60 min to various concentrations (from 0.1 to 10 M) of the xenobiotics. This species had previously proved to be a good bioindicator of organotin compounds. Both compounds, at concentrations of 1 and 10 M, altered the morphology of phagocytes, and these changes were closely related to disrupting effects on cytoskeletal components. At the same concentrations, phagocytosis, which requires cytoskeletal modifications for pseudopod formation, was severely hindered. Both compounds were able to induce apoptosis of Botryllus blood cells, probably as a consequence of severe oxidative stress related to the reported decrease of intracellular reduced glutathione (GSH) content. In the case of Sea-Nine 211 TM , a substantial increase in intracellular Ca 2+ and a negative effect on Ca 2+-ATPase activity may also be involved in the activation of the cell death machinery. Cytochrome-c-oxidase was also significantly inhibited by the two biocides, indicating perturbation of the mitochondrial respiratory chain. Isodynamic mixtures of Sea-Nine 211 TM and chlorothalonil were used to evaluate the occurrence of interactions between the two compounds. Results suggest the combined action of partial additivity when cell-spreading and cytochrome-c-oxidase activity were considered, and were indicative of antagonism in the case of the GSH depletion. On the whole, our results indicate that short-term in vitro exposure of haemocytes to high concentrations of Sea-Nine 211 TM and chlorothalonil provokes a marked reduction in haemocyte functionality, higher than or comparable to that of TBT. These assays of acute toxicity stress the immunosuppressive potential of these compounds, which, although counterbalanced by their short half-life in the marine environment, can lead to biocoenosis dismantling through rapid bioaccumulation by filter-feeding non-target benthic organisms.

The purpose of this study was to discuss the effect of the chemical structure of anionic and non-ionic surfactants and surface activity on toxicity. Single and binary mixtures of three ether carboxylic derivative surfactants and three amine-oxide-based non-ionic surfactants were used. Toxicity was determined using three test organisms: freshwater crustaceans (Daphnia magna), luminescent bacteria (Vibrio fischeri), and microalgae (Selenastrum capricornutum). The toxicity of surfactants is related to the hydrophobic alkyl chain, the degree of ethoxylation, and the critical micelle concentration of surfactants. Relationships found agreed with the fact that the lower toxicity is shown by the shorter alkyl chain. There is a strong relation between surface activity and toxicity: the toxicity increased as the CMC of the surfactant or mixtures of surfactants decreased. Commercial products are formulated using surfactants mixtures, so it is important to know their behavior using an easily measured property: the least toxic mixtures were formed by the surfactants having lower individual toxicity. Around the CMC, our data show a synergism for the binary mixtures. The results have given rise to a classification of the different surfactants and their mixtures according to the organism test, as safe, harmful or toxic. V. fischeri was in general the most sensitive microorganism to the toxic effect of the surfactants, followed by Daphnia magna, while Selenastrum capricornutum was more tolerant. These results can be useful for selecting technically efficient surfactants and their mixtures with a lower ecotoxicity on the aquatic environment.

A variety of chemicals including foaming agents are used to aid in the protection of forest resources from the wild land fires. The fire-fighting foams (FFFs) are formulations composed principally of surfactants. As a part of Japanese national project, environmentally inert FFFs have been developed. There is an emerging need for testing the impacts of these FFFs on the living organisms composing the typical landscapes or ecosystems in Japan. In the present report, we aimed to describe our latest attempt for assessing both the short-tem and long-term impacts of FFFs in aquatic (fresh water) and semi-aquatic (wetland) ecosystems , by employing the laboratory-sized model assays (preliminary studies) and the biotope-based observation, respectively. In the biotope assays which was based on the observation in compact biotopes mimicking the fresh water environments and wetland, both the acute and long-term eco-toxic impacts of two FFFs (soap-based and synthetic detergent-based) were assessed. Spraying of synthetic detergent-based foam formula was shown to be more toxic compared to soap-based formula and mock water treatment.

Cerium oxide nanoparticles (CeO2 NP) are increasingly used in industrial applications and may be released to the aquatic environment. The fate of CeO2 NP and effects on algae are largely unknown. In this study, the short term effects of CeO2 NP in two different agglomeration states on the green algae Chlamydomonas reinhardtii were examined. The role of dissolved cerium(III) on toxicity, its speciation and the dissolution of CeO2 NP were considered. The role of cell wall of C. reinhardtii as a barrier and its influence on the sensitivity to CeO2 NP and cerium(III) was evaluated by testing both, the wild type and the cell wall free mutant of C. reinhardtii. Characterization showed that CeO2 NP had a surface charge of ∼0mV at physiological pH and agglomerated in exposure media. Phosphate stabilized CeO2 NP at pH 7.5 over 24h. This effect was exploited to test CeO2 NP dispersed in phosphate with a mean size of 140nm and agglomerated in absence of phosphate with a mean size of 2000nm. The level of dissolved cerium(III) in CeO2 NP suspensions was very low and between 0.1 and 27nM in all tested media. Exposure of C. reinhardtii to Ce(NO3)3 decreased the photosynthetic yield in a concentration dependent manner with EC50 of 7.5±0.84μM for wild type and EC50 of 6.3±0.53μM for the cell wall free mutant. The intracellular level of reactive oxygen species (ROS) increased upon exposure to Ce(NO3)3 with effective concentrations similar to those inhibiting photosynthesis. The agglomerated CeO2 NP caused a slight decrease of photosynthetic yield at the highest concentrations (100μM), while no effect was observed for dispersed CeO2 NP. The low toxicity of agglomerated CeO2 NP was attributed quantitatively to Ce(3+) ions co-occurring in the nanoparticle suspension whereas for dispersed CeO2 NP, dissolved Ce(3+) was precipitated with phosphate and not bioavailable. Furthermore CeO2 NP did not affect the intracellular ROS level. The cell wall free mutant and wild type of C. reinhardtii showed the same sensitivity to CeO2 NP and Ce(NO3)3, indicating a minor role of the cell wall on toxicity. For both algae strains, a flocculation of cells was observed upon exposure to agglomerated CeO2 NP and Ce(NO3)3, only algae exposed to agglomerated CeO2 NP were tightly packed in exopolymeric substances.

Although toxic responses of freshwater
planarians after exposure to environmental toxicants can be
observed through external toxicological end points, physiological
responses inside the bodies of treated planarians
have rarely been investigated. The present study was
designed, using cadmium (Cd) as a reference toxicant, to
determine its bioaccumulation and toxicodynamics in the
freshwater planarian, Dugesia japonica, after acute toxicity
was obtained. Accumulated Cd concentrations, metallothionein
levels, and the oxidative status in planarians were
determined after exposure to Cd. Furthermore, we
hypothesized that the acute death of Cd-treated planarians
was associated with increased oxidative stress. After
Cd-treated planarians were coexposed to antioxidant,
N-acetylcysteine (NAC), we found that NAC protected
planarians from Cd lethality by maintaining the oxidative
status and decreasing the bioaccumulation of Cd. The
results of the present study support planarians being used
as a practical model for toxicological studies of environmental
contaminants in the future.

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Two bed media were tested (gravel and Filtralite) in shallow horizontal subsurface flow (HSSF) constructed wetlands in order to evaluate the removal of ammonia and nitrate for different types of wastewater (acetate-based and domestic wastewater) and different COD/N ratios. The use of Filtralite allowed both higher mass removal rates (1.1 g NH4–N m2 d1 and 3 g NO3–N m2 d1) and removal efficiencies (&gt;62% for ammonia, 90–100% for nitrate), in less than 2 weeks, when compared to the ones observed with gravel. The COD/N ratio seems to have no significant influence on nitrate removal and the removal of both ammonia and nitrate seems to have involved not only the conventional pathways of nitrification–denitrification. The nitrogen loading rate of both ammonia (0.8–2.4 g NH4–N m2 d1) and nitrate (0.6–3.2 g NO3–N m2 d1) seem to have influenced the respective removal rates.

In order to mimic the biological effects of an oil spill in Arctic waters, we examined several types of biomarkers (genes, enzymes, metabolites, and DNA damage) in polar cod Boreogadus saida experimentally exposed to the water soluble fractions of crude oil. During 4 weeks of ...

Nuclear magnetic resonance (NMR) based metabolomics in combination with multivariate data analysis may become valuable tools to study environmental effects of pharmaceuticals and other chemicals in aquatic organisms. To explore the usefulness of this approach in fish, we have used 1 H NMR metabolomics to compare blood plasma and plasma lipid extracts from rainbow trout exposed to the synthetic contraceptive estrogen ethinylestradiol (EE 2 ) with plasma from control fish. The plasma metabolite profile was affected in fish exposed to 10 ng/L but not 0.87 ng/L of EE 2 , which was in agreement with an induced vitellogenin synthesis in the high dose group only, as measured by ELISA. The main affected metabolites were vitellogenin, alanine, phospholipids and cholesterol. The responses identified by this discovery-driven method could be put in context with previous knowledge of the effects of estrogens on fish. This adds confidence to the approach of using NMR metabolomics to identify environmental effects of pharmaceuticals and other contaminants.

Pharmaceutically active compounds are produced worldwide and consumed in large
quantities, so these chemicals are frequently detected in limnic environments posing
potential ecological risks. Thirty pharmaceutically active compounds were selected
for examination of their acute toxicity for freshwater planarians (Dugesia japonica).
Among the 30 compounds tested, diclofenac, mefenamic acid, naproxen, propranolol
HCl, and diphenhydramine HCl had a 48-h nominal LC50 below10 mg L

1, and for
18 chemicals, it exceeded 100 mg L

1. The 96-h nominal LC50 was below 10 mg L

1
for diclofenac, mefenamic acid, naproxen, propranolol HCl, diltiazem HCl,
diphenhydramine HCl, hydroxyzine HCl, and triprolidine HCl, and for 15 chemicals,
it exceeded 100 mg L

1. Among different therapeutic groups, analgesics were most
toxic to planarians, and antibiotics were least toxic. Antihistamines and beta blockers
varied in their acute toxicity to planarians. At the current environmental levels, none
of the tested pharmaceutically active compounds may have acutely harmful impacts
on aquatic invertebrates. To answer the question whether chronic, long-term exposure
to pharmaceutically active compounds may entail ecological risks for aquatic
ecosystems, further investigations with different end points in multiple species tests
are needed.

Recognising the scientific and regulatory need for testing relatively hydrophobic or 'difficult substances', the OECD currently recommends that selected organic solvents may be used in aquatic toxicity testing in order to help achieve more effective dispersion of the toxicant. The OECD recommends a maximum solvent concentration of 100 l l −1 (with specific gravity equivalents to 100 l l −1 in parentheses) for acetone (79 mg l −1 ), dimethylformamide (95 mg l −1 ), dimethylsulfoxide (1.10 mg l −1 ), ethanol (78.9 mg l −1 ), methanol (79.2 mg l −1 ) and triethylene glycol (1.12 mg l −1 ). While this recommendation is supported by historical data, we have recently observed evidence that some solvents may affect the reproduction of certain fish species, and also impact biomarkers of endocrine disruption. This review presents available data on the effects of solvents in aquatic organisms, supplemented by relevant information from mammalian studies (e.g. effects on liver enzyme induction potentially altering the metabolism of sex hormones). In conclusion, it is recommended that maximum effort should be given to avoiding the use of carrier solvents wherever possible, for example through the use of saturation columns or other physical methods (e.g. stirring or ultrasonification). Where solvent use is necessary, however, it is recommended that in reproduction studies with aquatic organisms, the maximum solvent concentration should not exceed 20 l l −1 of dilution water.

There is an increasing body of evidence to support the significant role of invertebrates in assessing impacts of environmental contaminants on marine ecosystems. Therefore, in recent years massive efforts have been directed to identify viable and ecologically relevant invertebrate toxicity testing models. Tigriopus, a harpacticoid copepod has a number of promising characteristics which make it a candidate worth consideration in such efforts. Tigriopus and other copepods are widely distributed and ecologically important organisms. Their position in marine food chains is very prominent, especially with regard to the transfer of energy. Copepods also play an important role in the transportation of aquatic pollutants across the food chains. In recent years there has been a phenomenal increase in the knowledge base of Tigriopus spp., particularly in the areas of their ecology, geophylogeny, genomics and their behavioural, biochemical and molecular responses following exposure to environmental stressors and chemicals. Sequences of a number of important marker genes have been studied in various Tigriopus spp., notably T. californicus and T. japonicus. These genes belong to normal biophysiological functions (e.g. electron transport system enzymes) as well as stress and toxic chemical exposure responses (heat shock protein 20, glutathione reductase, glutathione S-transferase). Recently, 40,740 expressed sequenced tags (ESTs) from T. japonicus, have been sequenced and of them, 5673 ESTs showed significant hits (E-value, >1.0E−05) to the red flour beetle Tribolium genome database. Metals and organic pollutants such as antifouling agents, pesticides, polycyclic aromatic hydrocarbons (PAH) and polychrlorinated biphenyls (PCB) have shown reproducible biological responses when tested in Tigriopus spp. Promising results have been obtained when Tigriopus was used for assessment of risk associated with exposure to endocrine-disrupting chemicals (EDCs). Application of environmental gene expression techniques has allowed evaluation of transcriptional changes in T. japonicus with the ultimate aim of understanding the mechanisms of action of environmental stressors. Through a better understanding of toxicological mechanisms, ecotoxicologists may use this ecologically relevant species in risk assessment studies in marine systems. The combination of uses as a whole-animal bioassay and gene expression studies indicate that Tigriopus may serve as an excellent tool to evaluate the impacts of marine pollution throughout the coastal region. The purpose of this review is to illustrate the potential of using Tigriopus to fulfill the niche as an important invertebrate marine model organism for ecotoxicology and environmental genomics. In addition, the knowledge gaps and areas for further studies have also been discussed.

The industrial use of nanoparticles is rapidly increasing, and this has given rise to concerns about potential biological impacts of engineered particles released into the environment. So far, relatively little is known about uptake, accumulation and responses to engineered nanoparticles by plants. In this study, the effects of alumina nanoparticles on growth, morphology and photosynthesis of Lemna minor were quantified. It was found that alumina nanoparticles substantially increase biomass accumulation of L. minor. Such a stimulatory effect of alumina nanoparticles on growth has not been reported previously. Enhanced biomass accumulation was paralleled by morphological adjustments such as increased root length and number of fronds per colony, and by increased photosynthetic efficiency. Metal nanoparticles have previously been shown to enhance the energy transfer efficiency of isolated reaction centres; therefore it is proposed that the mechanism underlying the alumina mediated enhancement of biomass accumulation in L. minor is associated with increased efficiencies in the light reactions of photosynthesis.

The commercially important freshwater prawn, Macrobrachium rosenbergii post larvae (PL), 1.5±0.2 cm and
0.1±0.03 g were subjected to static renewal type acute toxicity bioassays against two orgnophophate insecticides,
quinalphos (Ekalux EC 25) and dimethoate 30% EC (TAFGOR). The 96 hr LC50 values were determined to be 0.774
μgl-1for quinalphos and 0.856 mgl-1for dimethoate. The PL were exposed to lethal (the 96 hr LC50) and sub-lethal (1/2nd and
1/4th of the 96 hr LC50) concentrations of these insecticides (quinalphos: 0.774, 0.384 and 0.193 μgl-1; dimethoate: 0.856,
0.428, 0.214 mgl-1) for a duration of 4, 8 and 12 days to study their acute and chronic impacts on whole body activities of
enzymes, the neurotransmitter, acetyl cholinesterase (AChE), the antioxidant, catalase and metabolic enzymes, glutamate
oxaloacetate transaminase (GOT) and glutamate pyruvate transaminase (GPT). The activity levels of AChE and catalase
were found to be significantly (P<0.05) decreased in test prawns when compared with control, whereas, there were significant
elevations in GOT and GPT levels (P<0.05). Among GOT and GPT, the impact was more on GOT than GPT. The
dosage and time dependent manner of inhibition or elevation in activities of these enzymes were recorded. Among these
two insecticides, quinalphos showed more impacts than dimethoate on this non-target organism.

Reclaimed water from small wastewater treatment facilities in the rural areas of the Beira Interior region (Portugal) may constitute an alternative water source for aquifer recharge. A 21-month monitoring period in a constructed wetland treatment system has shown that 21,500 m3 year−1 of treated wastewater
(reclaimed water) could be used for aquifer recharge. A GIS-based multi-criteria analysis was performed, combining ten thematic maps and economic, environmental and technical criteria, in order to produce a suitability map for the location of sites for reclaimed water infiltration. The areas chosen for aquifer
recharge with infiltration basins are mainly composed of anthrosol with more than 1 m deep and fine sand texture, which allows an average infiltration velocity of up to 1 m d−1. These characteristics will provide a final polishing treatment of the reclaimed water after infiltration (soil aquifer treatment (SAT)),
suitable for the removal of the residual load (trace organics, nutrients, heavy metals and pathogens).
The risk of groundwater contamination is low since the water table in the anthrosol areas ranges from 10 m to 50 m. On the other hand, these depths allow a guaranteed unsaturated area suitable for SAT. An area of 13,944 ha was selected for study, but only 1607 ha are suitable for reclaimed water infiltration.
Approximately 1280 m2 were considered enough to set up 4 infiltration basins to work in flooding and
drying cycles.

Variations in cytochrome P450 enzyme (CYPs) distribution and function between animal groups could result in differential metabolism and elimination kinetics for certain contaminants. Although a number of studies have suggested that differences in polychlorobiphenyl (PCB) accumulation profiles between crustacea and fish might result from differential CYP patterns, the relationship between PCB bioaccumulation and CYP capacities has not been demonstrated in these organisms. In the present study we investigated the hepatic microsomal catalytic activities in three deep-sea fish species, Alepocephalus rostratus (Alepocephalidae), Coelorinchus mediterraneus (Macrouridae), and Lepidion lepidion (Moridae), and the decapod crustacean Aristeus antennatus (Decapoda), using six fluorescent CYP-mediated substrates, namely ER (7-ethoxyresorufin), PR (7-pentoxyresorufin), BR (7-benzyloxyresorufin), CEC (3-cyano-7-ethoxycoumarin), DBF (dibenzylfluorescein) and BFC (7-benzyloxy-4-trifluoromethylcoumarin). Furthermore, we related the metabolic activities to the accumulation patterns of 41 PCB congeners in the muscle of these organisms. The results indicated a marked difference in the presence and activities of CYP isoforms between fish and the crustacean A. antennatus. Liver microsomes of the three selected fish species were capable of metabolizing all six CYP-mediated substrates and enzymes were identified as primarily belonging to CYP1A and CYP3A subfamilies. In contrast, hepatopancreas microsomes from A. antennatus only showed activity for PR and DBF substrates, generally related to mammalian CYP2-like enzymes. Furthermore, a direct relationship between metabolic activities and PCB accumulation profiles could be established. Results revealed that A. antennatus accumulated significantly higher proportions of PCBs 28, 52, 118, 138, 158 and 169 than fish, which is in accordance with the previously observed lack of CYP1A-like biotransformation capacities. Moreover, A. antennatus exhibited lower levels of PCBs 87, 149, 153, 170, 180, 183, 194 and 206 indicating that this crustacean is able to metabolize congeners considered mammalian CYP2B inducers. Hence, the present findings highlight the role of CYP-mediated metabolism in the congener-specific accumulation of PCBs in aquatic organisms and stress the need to further investigate quantitative and qualitative differences in xenobiotic metabolism among animal groups.

Acute toxicity tests of five metals (aluminum, cadmium, iron, lead, zinc) were performed to determine LC50 values in two
species of freshwater rotifers: Asplanchna brigthwellii and its prey Brachionus calyciflorus. We conducted the tests
using neonates less than 24 hr-old, each test consisted of five replicates, negative control and five metal concentrations
(Al, Cd, Fe, Pb, Zn). We found that the prey rotifer B. calyciflorus was more sensitive to Al, Cd, Pb and Fe than the
predator rotifer A. brightwellii. For both rotifers Cd was the most toxic of the five metals. Our B. calyciflorus strain is
sensitive when compared with other B. calyciflorus strains and other species and genera of the family Brachionidae. In
the other hand, LC50 values of A. brigthwellii are compared with rotifer and copepod predators.

Female three-spined sticklebacks were fed with freeze-dried chironomids contaminated with low or high doses of polybrominated diphenyl ethers (PBDEs) (Bromkal 70-5DE), polychlorinated naphthalenes (PCNs) (Halowax 1014) and polychlorinated biphenyls (PCBs) (Clophen A50). After 3.5 months of exposure, reproduction studies were started. No significant difference in number of eggs laid was found between the control and exposed groups. Spawning success (considered successful if it occurred within 24 h) in the control group was 80%, whereas it was 20% and 25% in the groups that received high doses of Bromkal 70-5DE or Clophen A50, respectively. After spawning, the fish were dissected for chemical, biochemical and morphological analyses. Levels of PBDE in the low-and high-dose Bromkal 70-5DE groups were 861 + 271 and 1630 + 275 mg/kg fat, respectively, whereas the corresponding concentrations of PCN in the Halowax 1014 groups were 845 + 43 and 1929 + 72 mg/kg fat, respectively. Concentrations of PCB in fish from the Clophen A50 groups were 1972 + 158 and 3594 + 521 mg/kg fat, respectively. Uptake efficiency was approx. 40% for PCB and 20% for PBDE and PCN. Hepatic cytochrome P450-dependent ethoxyresorufin-O-deethylase (EROD) activity was induced by Clophen A50 and Halowax 1014, whereas Bromkal 70-5DE did not significantly induce the enzyme system. EROD activity was 0.8 pmol/mg prot./min in the control group, whereas it was between 2.7 and 16.3 pmol/mg prot./min in the exposed groups. Furthermore, the morphological examination of the liver disclosed pronounced lipid accumulation in all exposed groups.

Bakteri Vibrio parahaemolyticus (Gambar 1) merupakan bakteri halofilik (hidup dengan kondisi kadar garam tinggi, hingga 30%). Habitat alami bakteri ini adalah di muara dan perairan laut pesisir. Bakteri ini memproduksi toksin hemolisin termostabil ekstraseluler yaitu "Thermostable Direct Haemolysin (TDH)" dan "Thermostable Direct Haemolysin-Related Haemolysin (TRH)". Kedua toksin ini masing-masing disandikan oleh gen virulensi tdh dan trh. Adanya gen tdh dan gen trh tersebut menentukan tingkat patogenitas strain V.

Antidepressants are among the most commonly detected human pharmaceuticals in the aquatic environment. Since their mode of action is by modulating the neurotransmitters serotonin, dopamine, and norepinephrine, aquatic invertebrates who possess transporters and receptors sensitive to activation by these pharmaceuticals are potentially affected by them. We review the various types of antidepressants, their occurrence and concentrations in aquatic environments, and the actions of neurohormones modulated by antidepressants in molluscs and crustaceans. Recent studies on the effects of antidepressants on these two important groups show that molluscan reproductive and locomotory systems are affected by antidepressants at environmentally relevant concentrations. In particular, antidepressants affect spawning and larval release in bivalves and disrupt locomotion and reduce fecundity in snails. In crustaceans, antidepressants affect freshwater amphipod activity patterns, marine amphipod photoand geotactic behavior, crayfish aggression, and daphnid reproduction and development. We note with interest the occurrence of non-monotonic dose responses curves in many studies on effects of antidepressants on aquatic animals, often with effects at low concentrations, but not at higher concentrations, and we suggest future experiments consider testing a broader range of concentrations. Furthermore, we consider invertebrate immune responses, genomic and transcriptomic sequencing of invertebrate genes, and the ever-present and overwhelming question of how contaminant mixtures could affect the action of neurohormones as topics for future study. In addressing the question, if antidepressants affect aquatic invertebrates at concentrations currently found in the environment, there is strong evidence to suggest the answer is yes. Furthermore, the examples highlighted in this review provide compelling evidence that the effects could be quite multifaceted across a variety of biological systems.

The present study was planned aiming to investigate the effects of atrazine
herbicide exposure on stress biomarkers acetylcholinesterase activity (AchE) and
cortisol, oxidative stress responses and the histopathological changes in liver of
female Clarias gariepinus and the ameliorative effect of vitamin E supplementation
(240mg/kg diet) on the degree of atrazine sublethal toxicity (1.37mg/l) for 6 weeks.
Chronic exposure to atrazine revealed a marked inhibition in the activity of AchE.
But, significant increase of serum cortisol level was recorded. Moreover, atrazine
exposure was associated with a marked induction of oxidative damage in liver tissue
as evidenced by increased level of lipid peroxidation (LPO) and reduced glutathione
(GSH) content. Atrazine exposure also led to a significant increase in the activities of
catalase (CAT) and superoxide dismutase (SOD).Furthermore, histopathological
examination of the liver of exposed fish showed dilatation and congestion of blood
vessels, fatty degeneration, necrosis and pyknotic nuclei of hepatocytes. However,
fish fed diet supplemented with vitamin E exhibited protective effect by minimizing
the atrazine-induced toxicity, through measured values more or less similar to the
control group fish.
Key words: Atrazine, Clarias gariepinus, Cortisol, Oxidative stress, Histopathology

There is an increasing body of evidence to support the significant role of invertebrates in assessing impacts of environmental contaminants on marine ecosystems. Therefore, in recent years massive efforts have been directed to identify viable and ecologically relevant invertebrate toxicity testing models. Tigriopus, a harpacticoid copepod has a number of promising characteristics which make it a candidate worth consideration in such efforts. Tigriopus and other copepods are widely distributed and ecologically important organisms. Their position in marine food chains is very prominent, especially with regard to the transfer of energy. Copepods also play an important role in the transportation of aquatic pollutants across the food chains. In recent years there has been a phenomenal increase in the knowledge base of Tigriopus spp., particularly in the areas of their ecology, geophylogeny, genomics and their behavioural, biochemical and molecular responses following exposure to environmental stressors and chemicals. Sequences of a number of important marker genes have been studied in various Tigriopus spp., notably T. californicus and T. japonicus. These genes belong to normal biophysiological functions (e.g. electron transport system enzymes) as well as stress and toxic chemical exposure responses (heat shock protein 20, glutathione reductase, glutathione S-transferase). Recently, 40,740 expressed sequenced tags (ESTs) from T. japonicus, have been sequenced and of them, 5673 ESTs showed significant hits (E-value, >1.0E−05) to the red flour beetle Tribolium genome database. Metals and organic pollutants such as antifouling agents, pesticides, polycyclic aromatic hydrocarbons (PAH) and polychrlorinated biphenyls (PCB) have shown reproducible biological responses when tested in Tigriopus spp. Promising results have been obtained when Tigriopus was used for assessment of risk associated with exposure to endocrine-disrupting chemicals (EDCs). Application of environmental gene expression techniques has allowed evaluation of transcriptional changes in T. japonicus with the ultimate aim of understanding the mechanisms of action of environmental stressors. Through a better understanding of toxicological mechanisms, ecotoxicologists may use this ecologically relevant species in risk assessment studies in marine systems. The combination of uses as a whole-animal bioassay and gene expression studies indicate that Tigriopus may serve as an excellent tool to evaluate the impacts of marine pollution throughout the coastal region. The purpose of this review is to illustrate the potential of using Tigriopus to fulfill the niche as an important invertebrate marine model organism for ecotoxicology and environmental genomics. In addition, the knowledge gaps and areas for further studies have also been discussed.

The use of alternative feeds aims to maintain productivity and reduce animal production costs. The objective of this study was to determine the nutritional value of Water Spinach (WS) (Ipomoea aquatica), as well as replacement of fishmeal (FM) by water spinach in diets for Nile tilapia fry, Oreochromis niloticus. To determine the growth performance of Nile tilapia fed different levels of water spinach, 150 fish (3.25±0.44g; 3.6±0.2cm) were randomly distributed in flow through system of 15 plastic aquariums each containing about 20 liters, and fed during 7 weeks with isoprotein (30% digestible protein) and isoenergetic (14.35 kJ g-1 digestible energy) diets containing replacement levels of 0, 25, 50, 75 and 100% of FM digestible protein by WS digestible protein. The experimental design was completely randomized with five treatments and three replicates. The protein efficiency ratio, absolute body weight, live weight gain and feed conversion ratio content significantly decreased in fish fed diets containing WS levels above 25%. Therefore, WS can replace up to 25% of FM without impairing juvenile Nile tilapia growth performance, feed efficiency, and body composition.

Pokok bahasan dalam perkuliahan mata kuliah ini terdisi atas Ruang Lingkup toksikologi perairan, nasib zat beracun dalam tubuh biota perairan, distribusi zat toksikan di air, sedimen dan biota perairan, aneka kondisi toksi, mekanisme aksi, wujud dan sifat efek toksik, tolok ukur toksisitas secara kualitatif dan kuantitatif, ragam uji toksisitas.

Organophosphorus pesticides are the most widely and commonly used insecticides worldwide. Toxicity and impact of Dichlorvos, an organophosphate pesticide polluting aquatic ecosystems as a potential toxicant was reviewed. This article summarized the LC 50 values of dichlorvos to various fish species and significant findings pertaining to its lethal and sublethal toxic effects in various aspects of ecotoxicological perturbations in fish which can be viewed as biomarkers of pesticide toxicity. These biomarkers reported due to toxic effect of the dichlorvos can be used to monitor pollution risk assessment in aquatic ecosystems.