The cerium oxide nanoparticles toxicity induced physiological, histological and biochemical alterations in freshwater mussels, Unio crassus.

INTRODUCTION: Releasing of cerium oxide nanoparticles (nano-CeO2) to the nature has increased due to the widespread use in many fields ranging from cosmetics to the food industry. Therefore, nano-CeO2 has been included in the Organization for Economic Co-operation and Development's (OECD) priority list for engineering nanomaterials. In this study, the effects of nano-CeO2 on the freshwater mussels were investigated to reveal the impact on the freshwater systems on model organism. METHODS: First, the chemical and structural properties of nano-CeO2 were characterized in details. Second, the freshwater mussels were exposed to environmentally relevant concentrations of nano-CeO2 as 10 mg, 25 mg and 50 mg/L during 48-h and 7-d. Third, after the exposure periods, hemolymph and tissue samples were taken to analyse the Total Hemocyte Counts (THCs) histology and oxidative stress parameters (total antioxidant status, glutathione, glutathione-S-transferase, and advanced oxidative protein products). RESULTS: Significant decrease of the THCs was observed in the nano-CeO2 exposed mussels compared to the control group (P < 0.05). The histological results showed a positive association between nano-CeO2 exposure concentration in the water and level of tissue damage and histopathological alterations were detected in the gill and the digestive gland tissues. Oxidative stress parameters were slightly affected after exposure to nano-CeO2 (P > 0.05). In conclusion, this study showed that acute exposure of freshwater mussels to nano-CeO2 did not pose significant biological risk. However, it has been proven that mussels are able to accumulate nano-CeO2 significantly in their bodies. CONCLUSION: This suggests that nano-CeO2 may be a potential risk to other organisms in the ecosystem through trophic transfer in the food-web based on their habitat and niche in the ecosystem.

Genotoxicity of sub-lethal di-n-butyl phthalate (DBP) in Nile tilapia (Oreochromis niloticus).

This study aimed to assess genotoxicity in Nile tilapia (Oreochromis niloticus) erythrocytes after exposure to a sub-lethal concentration of 10 mg L(-1) di-n-butyl phthalate (DBP) for 24 and 96 h. The results showed that mean MN frequencies in both DBP and ethyl methane-sulfonate (EMS, positive control for MN bioassay) groups were significantly different (p<0.01) with respect to control and solvent control groups, in both exposure scenarios. When analysing nuclear abnormalities, the frequency of notched nuclei was significantly different (p<0.05) but the frequencies of other subtypes did not change. The 96-h exposure led to an increase in the mean frequencies of notched nuclei, and also caused significant differences between MN frequencies in all groups (p<0.01). Our findings indicate that sub-lethal DBP concentrations when tested in controlled laboratory conditions have genotoxic potential towards Nile tilapia. Further detailed studies should be done for the determination of the environmental risk assessment for aquatic life since DBP is a high risk contaminant of freshwater and marine ecosystems.

Impact of DBP on histology and expression of HSP 70 in gill and liver tissue of Cyprinus carpio.

Di-n-butyl phthalate (DBP) widely used plasticizer in the plastic industry, affects regulation of the endocrine system and causes toxicity in animals. In the present study, the aim was to study the toxic effects/damages of DBP exposure using Hsp70 levels and histopathological changes in Carp liver and gill. Hsp70 expression levels were assessed as specific biomarker of in vivo ecotoxicological stress. Carp (Cyprinus carpio) were exposed to sub-lethal concentration of DBP (di-n-butyl phthalate, 1 mg/L) for 4, 24 and 96 h. Gill and liver tissues were evaluated histopathologically and RNA quantifications for Hsp70 expression levels were carried out using a two-step real-time RT-PCR. In liver, a rapid but non-significant increase in mRNA levels in the first 4 h was observed. mRNA levels significantly increased up to 2-3 fold after 24 and 96 h (p < 0.05). However, irregular mRNA level changes were also recorded: Gill specific and time-dependent regulation of Hsp70 expression were 4-5 fold inhibition after 4 and 24 h (p < 0.05), then increased up to 4 fold after 96 h (p < 0.05). Histopathological findings support altered transcription results as: Epithelial lifting, hyperplasia, fusion of secondary lamellae, telangiectasis, passive hyperemia and hydropic degeneration. Significant alterations of Hsp70 levels were likely due to a tissue specific response against chemical stress, cellular damage and lesions due to DBP. Carp was found to be a suitable experimental model for toxicology, and Hsp70 mRNA levels are reliable, specific biomarkers.

Toxicity and molecular effects of di-n-butyl phthalate (DBP) on CYP1A, SOD, and GPx in Cyprinus carpio (common carp).

Di-n-butyl phthalate (DBP), a widely used plasticizer in the plastic industry, affects regulation of the endocrine system and causes toxicity in animals. In the present study, we evaluated a series of ecotoxicological stress biomarkers in the common carp (Cyprinus carpio) as an experimental model to test for alterations in gene expression at a sublethal concentration of 1 mg/L DBP for 4, 24, and 96 h. In gills, an immediate increase in CYP1A messenger RNA (mRNA) levels was observed within the first 4 h and persisted for 96 h. Protein levels were nearly consistent with mRNA levels. However, a time-dependent inhibition was observed in CYP1A levels in the liver within 96 h. Superoxide dismutase (SOD) and glutathione peroxidase (GPx) levels increased gradually in liver with exposure time to a maximum level of 11-fold. Varied responses of different tissues were likely due to xenobiotic metabolism of DBP. In conclusion, evaluating the tissue-specific alterations of CYP1A, SOD, and GPx levels can be used as specific and effective biomarkers for ecotoxicological monitoring of DBP pollution. We strongly recommend using molecular tools to ecotoxicologists for aquatic monitoring of newly emerging pollutants.

Sublethal toxicity of esbiothrin relationship with total antioxidant status and in vivo genotoxicity assessment in fish (Cyprinus carpio L., 1758) using the micronucleus test and comet assay.

Esbiothrin, synthetic pyrethroid with quick activity against insects, is widely used against household pests and in public health. Despite widespread use, data on ecotoxicity and genotoxic effects are extremely scarce. The aim of the present study is to evaluate the genotoxic potential of esbiothrin on a model fish species Cyprinus carpio L., 1758 (Pisces: Cyprinidae, koi) using the micronucleus test and comet assay in peripheral blood erythrocytes. Effects of two sublethal exposure concentrations on plasma total antioxidant status (TAS mmol/L), and Hct values were examined. On the basis of the 96 h LC50 data from U.S. EPA ecotox database (32 µg/L) two sublethal exposure concentrations (5 and 10 µg/L) were used together with ethyl methanesulfonate (EMS) (5 mg/L) as positive control. Five fish were used for each dose/duration group (24, 48, and 72 h) under controlled laboratory conditions. The fish showed behavioral changes at the higher dose. Plasma TAS (mmol/L) levels decreased in 24 h; an increase was observed slightly for 48 and obviously for 72 h in both exposure doses. Similarly, hematocrit (Hct) values differed between exposure duration but no significant differences in mean values were found between groups of the same exposure time. The general trend was a rise after 48 h, which decreased afterwards. Our results revealed significant increases in the frequencies of micronuclei and levels of DNA strand breaks and thus demonstrated the genotoxic potential of this pesticide on fish, a nontarget organism of the aquatic ecosystem. To our knowledge this is the first study to report observable genotoxic effects of esbiothrin on fish.

Sublethal propoxur toxicity to juvenile common carp (Cyprinus carpio L., 1758): biochemical, hematological, histopathological, and genotoxicity effects.

The sublethal toxicological and genotoxic potential of propoxur, a widely used carbamate insecticide against household pests, in veterinary medicine, and in public health, was evaluated on carp as a model species (Cyprinus carpio L., 1758) using the erythrocyte micronucleus test. Based on the 96-h lethal concentration, 50% (LC50) data from the U.S. Environmental Protection Agency ECOTOX Database (10 mg/L), a sublethal exposure concentration of 5 mg/L was used under static bioassay laboratory conditions. Histopathological evaluation showed no significant changes in spleen, intestine, muscle, or skin tissues. However, the following conditions were recorded: hyperemia, branchitis in primary lamella, and telangiectasis, hyperplasia, fusion, epithelial lifting, and epithelial desquamation in secondary lamella of gill tissues; hemorrhage, destruction, prenephritis, and inflammation and desquamation in the tubules; edema in the kidney; passive hyperemia, albumin, and hydropic degeneration in the liver; and hyperemia, chromatolysis, and glial proliferation in brain tissues. No statistically significant increases in micronuclei frequencies were found. Hematological parameters showed decreased hematocrit values and mean corpuscular volume values, as well as increased erythrocyte and leukocyte counts compared with the control group (p < 0.01). Plasma glucose, cholesterol, triglycerides, phosphorus, sodium, total plasma protein, chloride, and aspartate aminotransferase levels were increased (p < 0.01). Only plasma calcium and potassium levels decreased in the experimental group. Propoxur has an ecotoxicological potential on fish, a nontarget organism.

Evaluation of potential genotoxicity of five food dyes using the somatic mutation and recombination test.

In this study, different concentrations of five food dyes (amaranth, patent blue, carminic acid, indigotine and erythrosine) have been evaluated for genotoxicity in the Somatic Mutation and Recombination Test (SMART) of Drosophila melanogaster. Standard cross was used in the experiment. Larvae including two linked recessive wing hair mutations were chronically fed at different concentrations of the test compounds in standard Drosophila Instant Medium. Feeding ended with pupation of the surviving larvae. Wings of the emerging adult flies were scored for the presence of spots of mutant cells which can result from either somatic mutation or somatic recombination. For the evaluation of genotoxic effects, the frequencies of spots per wing in the treated series were compared to the control group, which was distilled water. The present study shows that carminic acid and indigotine demonstrated negative results while erythrosine demonstrated inconclusive results. In addition 25 mg mL(-1) concentration of patent blue and 12.5, 25 and 50 mg mL(-1) concentrations of amaranth demonstrated positive results in the SMART.

Genotoxicity assessment of carp (Cyprinus carpio L.) fingerlings by tissue DNA damage and micronucleus test, after environmental exposure to fenitrothion.

The purpose of this study was to evaluate the adverse effects of sublethal doses of fenitrothion, an organophosphothionate insecticide on brain, gill, liver, and muscle tissues as a ratio of 8-OHdG to dG to indicate the DNA damage and erythrocyte micronucleus frequency for genotoxicity of carp (Cyprinus carpio L.) fingerlings. In our study, the mean weights and lengths of the fish (n = 4-12) were 31.13 ± 14.24 g and 12.53 ± 1.41, respectively. Before the experiment, fish were maintained in aerated dechlorinated tap water at 21.8 ± 1 °C and fed daily with commercial feed at a rate of 2% of their body weights. Experiments were conducted under static conditions in the aquaria. Technical grade (95%) fenitrothion was diluted in acetone to give a dosing solution of 10 mg/L. The increased lesions/106 DNA bases (p < 0.05) of liver tissue of exposure group (0.49 ± 0.18) was observed when compared to control group (0.28 ± 0.30). There was not any significant differences between brain tissues, no damage were detectable in gill and muscle tissues of control groups, and in exposure groups altered levels of damage were detected for gill (0.06 ± 0.05) and muscle (0.16 ± 0.21) tissues. The increased micronucleus frequencies (%) in erythrocytes of carp following the exposure to 48 h fenitrothion (6.43 ± 3.89; p<0.05) was observed when compared to control group (1.29 ± 1.03). The available data indicate that there is still lack of well-established dose-response relationships between occupational or environmental exposures and the induction of 8-OHdG. Such biomarkers may be used in assessing adverse/toxic effects of pesticides as environmental stressors.

The acute toxicity of fenitrothion on narrow-clawed crayfish (Astacus leptodactylus Eschscholtz, 1823) in association with biomarkers of lipid peroxidation.

The aim of this research was to evaluate the acute toxicity of fenitrothion to the crayfish (Astacus leptodactylus Eschscholtz, 1823), which is chosen as an alternative aquatic organism to fish by using the static test system and evaluate the basic lipid peroxidation parameters for the first 24 h. Crayfish of 27.3 ± 0.56 g mean weight and 10.0 ± 0.72 cm mean length were selected for the bioassay experiments. The experiments were repeated three times in 20 liters of tap water. The temperature of water was 21 ± 1°C. The data obtained were statistically evaluated by using a computer program developed by the United States Environmental Protection Agency, based on Finney's probit analysis method and the 96-h LC(50) value for crayfish was calculated to be 15.75 µg/L. The 95% lower and upper confidence limits for the LC(50) were 9.45 to 25.01 µg/L. In addition to the acute toxicity bioassay experiments, 24-h oxidative stress parameters such as malondialdehyde (MDA) levels and ferrous oxidation assay (FOX HP [hydrogen peroxide] equivalents) were also determined. Only MDA levels of hepatopancreas decreased at 5, 10, and 20 µg/L of fenitrothion doses. We can conclude that fenitrothion is highly toxic to crayfish, a nontarget organism in the ecosystem, and the lipid peroxidation indicators can be easily used for monitoring environmental effects.

Quantum chemical treatment of beta-sitosterol molecule.

Beta-sitosterol is used as a dietary supplement for lowering plasma cholesterol, and has atherosclerosis preventive, anti-inflammatory, antimicrobial, antipyretic, induced apoptosis, and anticancer effects. In order to understand the effect of the molecule we have investigated the molecule theoretically. The structural, vibrational, and electronic properties of the beta-sitosterol molecule have been investigated theoretically by performing molecular mechanics (MM+ force field), semiempirical self-consistent-field molecular-orbital (PM3), and density functional theory (B3LYP) calculations. The geometry of the considered molecule has been optimized; the vibrational dynamics and the electronic properties have been calculated in its ground state in the gas phase. The optimized structure of the molecule is not planar, and its heat of formation is exothermic. The calculated infrared spectrum for beta-sitosterol agrees well qualitatively with the experimentally determined FTIR spectrum. The interfrontier molecular orbitals are localized mainly on the double C-C bond, and the energy difference of the corresponding orbitals is relatively small, which makes the molecule kinetically stable. According to the calculated dipole moment, beta-sitosterol is a polar molecule. The calculated results for the beta-sitosterol molecule in the present study will aid in elucidation of the mechanism of action and may further be used in lipid metabolism drug design studies.

Quantum chemical investigation of nitrotyrosine (3-nitro-L-tyrosine) and 8-nitroguanine.

The structural, vibrational and electronic properties of nitrotyrosine and 8-nitroguanine have been investigated theoretically by performing the molecular mechanics (MM+ force field), the semi-empirical self-consistent-field molecular-orbital (PM3), and density functional theory calculations. The geometry of the nitrotyrosine and 8-nitroguanine molecules have been optimized, the vibrational dynamics and the electronic properties calculated in their ground states in the gas phase.

Sublethal cyfluthrin toxicity to carp (Cyprinus carpio L.) fingerlings: biochemical, hematological, histopathological alterations.

This study measures a whole series of biological endpoints, and histopathological and behavioral changes of carp (Cyprinus carpio L.) fingerlings in order to investigate the adverse effects of a sublethal concentration of 10 microg/L cyfluthrin for 48h and one week using the semi-static test system. The most significantly altered findings of this study was that total antioxidant status levels decreased in 48h, chloride levels decreased in one week, and sodium and phosphorous levels increased in the 48h-exposed group. Plasma cortisol increased in both the 48h-exposed and the one-week-exposed groups. The unique tissue was the brain that had an increased level of malondialdehyde after one week of exposure. Such biomarkers may be used in assessing adverse/toxic effects of pesticides as environmental stressors. Potential oxidative-stress-inducing effect of synthetic pyrethroid cyfluthrin, as lipid peroxidation, in addition to its sodium channel blocking, is reported for the first time as an additional mechanism of action.

Investigation of acute toxicity of fenitrothion on guppies Poecilia reticulata.

Fenitrothion, an organophosphothionate insecticide (CAS number: 122-14-5) and potential toxic pollutant contaminating aquatic ecosystems, was investigated in the present study for acute toxicity. Guppy fish (Poecilia reticulata) were selected for the bioassay experiments. The experiments were repeated three times and the 96 h LC(50) was determined for the guppies. The static test method of acute toxicity test was used. The water temperature was regulated at 23 +/- 1 degrees C. In addition, behavioral changes at each fenitrothion concentration were observed for the individual fish. Data obtained from the fenitrothion acute toxicity tests were evaluated using the probit analysis statistical method. The 96 h LC(50) value for guppy was estimated as 3.28 mg l(-1). Values in the range of microg l(-1) and mg l(-1) have been reported for various other fish species.

Acute toxicity, behavioral changes, and histopathological effects of deltamethrin on tissues (gills, liver, brain, spleen, kidney, muscle, skin) of Nile tilapia (Oreochromis niloticus L.) fingerlings.

Deltamethrin, a synthetic pyrethroid contaminating aquatic ecosystems as a potential toxic pollutant, was investigated in the present study for acute toxicity. The purpose of this study was to evaluate LC(50) values of deltamethrin on Nile tilapia (Oreochromis niloticus L.) fingerlings and investigate histopathological responses of fish exposed to deltamethrin. The 48 h LC(50) value for Nile tilapia fingerlings was estimated as 4.85 microg/L using static test system. In addition, behavioral changes at each deltamethrin concentration were observed closely. All fish, exposed to 5 microg/L deltamethrin revealed severe morphological alterations in the gills and liver. In the gills hyperemia, fusion of secondary lamellae and telangiectasis were observed; whereas hydropic degenerations in liver were observed in all examined fish. The results are significant for reporting acute deltamethrin toxicity in terms of behavioral and histopathological changes: Deltamethrin is highly toxic to fingerlings.

Investigation of acute toxicity of chlorpyrifos-methyl on guppy Poecilia reticulata.

Static bioassays were made to determine acute toxicity of chlorpyrifos-methyl, a wide spectrum organophosphorus insecticide and potential toxic pollutant of aquatic ecosystem, Guppy fish (Poecilia reticulata). Bioassays were made at a regulated temperature of 22+/-1 degrees C and were repeated three times. Lethal doses of the insecticides were determined using LC50 software programme of U.S. EPA based on Finney's Probit Analysis statistical method. The 96 h LC50 value and 95% confidence limit of chlorpyrifos-methyl for Guppy was estimated as 1.79 (1.47-2.10) mg/l. The fish exposed to chlorpyrifos-methyl exhibited behavioral changes in the form of neurotoxin toxicity: less general activity than control group, loss of equilibrium, erratic swimming and staying motionless at a certain location generally at mid-water level for prolonged periods. The 1 mg/l (lowest) concentration had similar behavior (NOEC) with the control group.

Investigation of acute toxicity of fenitrothion on peppered corydoras (Corydoras paleatus) (Jenyns, 1842).

Fenitrothion, as an organophosphothionate insecticide, is a contact insecticide and selective acaricide, also used as a vector control agent for malaria in public health programs. A 96 h LC50 value of fenitrothion, a potential toxic pollutant contaminating aquatic ecosystems, was determined on the adult peppered corydoras (Corydoras paleatus). The experiments were repeated three times. The static test method of acute toxicity test was used. Water temperature was regulated at 23 +/- 1 degrees C. In addition, behavioral changes at each fenitrothion concentration were observed for the individual fish. Data obtained from acute toxicity tests were evaluated using the Probit Analysis Statistical Method. The 96 h LC50 value for peppered corydoras was estimated as 3.51 mg/l.

Investigation of acute toxicity of deltamethrin on guppies (Poecilia reticulata).

Deltamethrin, a synthetic pyrethroid pesticide contaminating aquatic ecosystems as a potential toxic pollutant, was investigated in the present study for acute toxicity. Guppy fish (Poecilia reticulata) were selected for the bioassay experiments. The 48-h LC50 was determined for the guppies. The experiments were repeated three times and a total of 210 guppies were used. The static test method of acute toxicity test was used. In addition, behavioral changes at each deltamethrin concentration were determined for the individuals. Data obtained from the deltamethrin acute toxicity tests were evaluated using the Probit Analysis Statistical Method. The 48-h LC50 value for guppy was estimated as 5.13 microg/L.

Investigation of acute toxicity of permethrin on guppies Poecilia reticulata.

Permethrin, a synthetic pyrethroid pesticide and potential toxic pollutant contaminating aquatic ecosystems, was investigated in the present study for acute toxicity. Guppy fish (Poecilia reticulata) were selected for the bioassay experiments. The experiments were repeated 3 times and the 48-h LC(50) was determined for the guppies. The static test method of acute toxicity test was used. Water temperature was regulated at 20+/-1 degrees C. In addition, behavioral changes at each permethrin concentration were observed for the individual fish. Data obtained from the permethrin acute toxicity tests were evaluated using the probit analysis statistical method. The 48-h LC(50) value for guppy was estimated as 245.7 microg/l. Values in the range of 0.05-97.0 microg/l have been reported for various other fish species.

Water quality and impacts of pollution sources for Eymir and Mogan Lakes (Turkey).

Mogan and Eymir Lakes are two shallow lakes, interconnected hydrologically in the close vicinity of Ankara, Turkey. A total of 245 km(2) of the total 971.4 km(2) watershed is under environmental protection status as "Gölbasi Specially Protected Area". Potential impacts from extensive agriculture, recreation, incomplete infrastructure and other human activities, such as residential settlements, are discussed with reference to previous and more recent pollution monitoring. Six monitoring stations enabling follow-up of previous work were selected in this study. These were on the creeks feeding the lake systems. Generally, summer months showed heavier pollution loads, with Eymir Lake concentrating the pollutants due to flow from Mogan Lake. When compared with the 1995 study; COD, total-P, Kjeldahl-N in the six stations were close or slightly decreased in the present study. Suspended solids significantly decreased; possibly due to erosion control measures and decreased domestic wastewater. The improvement in the pollution state of the lakes is attributed to the construction of a sewage system going around Mogan Lake and collecting wastewater discharges and restrictions to urban settlement development around the lakes brought by the 1/25,000 land use plan controlling further impact from residential developments within the protected area boundaries. The study, while addressing water quality and interactions due to human activities in shallow lakes, also discusses problems associated with human impacts in protected areas with the aim of presenting a complicated case study.

Investigation of acute toxicity of beta-cypermethrin on guppies Poecilia reticulata.

Beta-cypermethrin, a synthetic pyrethroid pesticide and potential toxic pollutant, contaminating aquatic ecosystems was investigated in the present study for acute toxicity. Guppy fish (Poecilia reticulata) was selected for the bioassay experiments. The experiments were repeated three times and the 48-h LC50 was determined for the guppies. The static test method of acute toxicity test was used. Water temperature was regulated at 22 +/- 1 degrees C. In addition, behavioral changes at each beta-cypermethrin concentration were observed for the individual fish. Data obtained from the beta-cypermethrin acute toxicity tests were evaluated using the probit analysis statistical method. The 48-h LC50 value for guppy was estimated as 21.4 microg/l.