Nano-titanium dioxide induced genotoxicity and histological lesions in a tropical fish model, Nile tilapia (Oreochromis niloticus).

This study evaluated potential genotoxic and histopathological effects of nano-TiO2 (0.1, 0.5 and 1 mg/L) in Nile tilapia over 7, 14 and 21 days of exposure. Bulk TiO2 (1 mg/L) along with controls was used for comparison. Comet assay revealed that nano-TiO2 can induce erythrocytic DNA damage in a concentration dependent manner. However, micronuclei induction was observed only at the lowest concentration. Elevated organ damage indices indicate nano-TiO2 induced histological alterations in liver and intestine. Severe histological alterations induced by nano-TiO2 in the fish were necrosis of hepatic parenchyma and intestinal mucosa. Bulk TiO2 exposure had no effect on the histological structure of the intestine but increased liver damage indices and erythrocytic DNA damage compared to the controls indicating dissolved form of TiO2 is not biologically inert. More research efforts are needed to generate in vivo toxicity data on realistic levels of nano-TiO2 and bulk TiO2 for environmental risk assessments.

Comparison of Sensitivity of Tropical Freshwater Microalgae to Environmentally Relevant Concentrations of Cadmium and Hexavalent Chromium in Three Types of Growth Media.

Sensitivity of tropical freshwater microalgae (Mesotaenium sp., Chlorococcum sp. and Scenedesmus sp.) to environmentally relevant concentrations of hexavalent chromium (Cr6+) and cadmium (Cd2+) was compared individually in three growth media viz. Bold's Basal Medium (BBM), Test Medium 1 (TM1) and Test Medium 2 (TM2) based on fluorescence reduction. Free metal content of growth media was determined by Visual MINTEQ (version 3.1). After 24 h, relative fluorescence of microalgae in the three media decreased with increased metal concentration showing a concentration dependent graded toxicity response. All microalgae were more sensitive to the metals when grown in TM1, when compared, more sensitive to Cr6+ than Cd2+. Metal speciation indicated that TM1 and TM2 media have higher percentage of bioavailable Cd2+ than BBM, and chromium was present mainly as CrO42- and HCrO4-. The results suggest that the TM1 medium is more suitable under short term exposure of microalgae to metals in environmental monitoring.

Bioassessment of the Effluents Discharged from Two Export Oriented Industrial Zones Located in Kelani River Basin, Sri Lanka Using Erythrocytic Responses of the Fish, Nile Tilapia (Oreochromis niloticus).

Complex effluents originating from diverse industrial processes in industrial zones could pose cytotoxic/genotoxic hazards to biota in the receiving ecosystems which cannot be revealed by conventional monitoring methods. This study assessed potential cytotoxicity/genotoxicity of treated effluents of two industrial zones which are discharged into Kelani river, Sri Lanka combining erythrocytic abnormality tests and comet assay of the tropical model fish, Nile tilapia. Exposure of fish to the effluents induced erythrocytic DNA damage and deformed erythrocytes with serrated membranes, vacuolations, nuclear buds and micronuclei showing cytotoxic/genotoxic hazards in all cases. Occasional exceedance of industrial effluent discharge regulatory limits was noted for color and lead which may have contributed to the observed cytotoxicity/genotoxicity of effluents. The results demonstrate that fish erythrocytic responses could be used as effective bioanalytical tools for cytotoxic/genotoxic hazard assessments of complex effluents of industrial zones for optimization of the waste treatment process in order to reduce biological impacts.

Sensitivity of Four Cyanobacterial Isolates from Tropical Freshwaters to Environmentally Realistic Concentrations of Cr(6+), Cd(2+) and Zn(2.).

Sensitivity of four tropical cyanobacteria viz. Coelosphaerium sp., Synechococcus sp., Oscillatoria sp. and Chroococcus sp. to environmentally relevant concentrations of Cr(6+), Cd(2+) and Zn(2+)was assessed based on fluorescence change as a proxy for growth reduction. At 24 h exposure, the growth reduction inthe cyanobacteria followed the order: Zn(2+) < Cr(6+) ≤ Cd(2+). Of the four cyanobacteria, Synechococcus was the most sensitive for Cr(6+), where as Chroococcus was the most sensitive for Cd(2+)and Zn(2+). Sensitivity was gradually decreased by 96 h implying the acquisition of tolerance by cyanobacteria to heavy metal ions with prolonged exposure.

Assessment of bile fluorescence patterns in a tropical fish, Nile tilapia (Oreochromis niloticus) exposed to naphthalene, phenanthrene, pyrene and chrysene using fixed wavelength fluorescence and synchronous fluorescence spectrometry.

Bile fluorescence patterns in Nile tilapia, a potential fish for biomonitoring tropical water pollution were assessed following exposure to selected polycyclic aromatic hydrocarbons (PAHs): naphthalene, phenanthrene, pyrene and chrysene. Non-normalized fixed wavelength fluorescence signals in the fish exposed to these PAHs reflected dose and/or time response relationships of their metabolism. Normalizing signals to biliverdin introduced deviations to these response patterns. The optimal wavelength pairs (excitation/emission) for synchronous fluorescence scanning measurements of bile metabolites of naphthalene, phenanthrene, pyrene and chrysene were identified as 284/326, 252/357, 340/382 and 273/382 respectively. This study supports the use of bile fluorescence in Nile tilapia by fixed wavelength fluorescence and synchronous fluorescence spectrometry with non-normalized data as a simple method for screening bioavailability of these PAHs.

Use of biomarkers in Nile tilapia (Oreochromis niloticus) to assess the impacts of pollution in Bolgoda Lake, an urban water body in Sri Lanka.

The present study reports the first analysis of water pollutants in Sri Lankan waters using a suite of biomarkers in Nile tilapia (Oreochromis niloticus) residing in Bolgoda Lake which receives urban, industrial and domestic wastes from multiple sources. The fish were collected from the lake in the dry period (April 2005) and wet periods (September 2005, October 2006) and the levels of biomarkers viz. hepatic ethoxyresorufin O-deethylase (EROD), glutathione S-transferase (GST), metallothioneins, biliary fluorescent aromatic compounds, brain and muscle cholinesterases (ChE) were compared with those of the laboratory reared control fish and the fish obtained from a less polluted water body, Bathalagoda reservoir (reference site). The results revealed that biomarker levels of the fish collected from the reference site were not significantly different from the controls. Hepatic EROD and GST activities in fish from Bolgoda Lake were induced 4.2-16.6 folds and 1.4-3.3 folds respectively compared with the control fish. Analysis of bile in the lake fish revealed recent uptake of naphthalene, pyrene and benzo(a)pyrene type polycyclic aromatic hydrocarbons (PAHs). The induction of EROD activities in feral fish reflects the exposure of fish to aryl hydrocarbon receptor agonists including PAHs present as pollutants in the Bolgoda Lake. Cholinesterase activity in the fish inhabiting one sampling site of Bolgoda Lake was lower (22-40% inhibition) than the activity measured in the control fish indicating the presence of anticholinesterase pollutants in the area. Hepatic metallothionein levels in the lake fish were higher (1.9-3.2 folds) in comparison to the controls indicating metal exposure. The results support the potential use of these biomarkers in Nile tilapia in assessing pollution in tropical water bodies.

Effects of biological and technical factors on brain and muscle cholinesterases in Nile tilapia, Oreochromis niloticus: implications for biomonitoring neurotoxic contaminations.

Influence of body length, body weight, gender, sexual maturity, and tissue storage on brain and muscle cholinesterases (ChE) in Nile tilapia was evaluated considering its potential use in biomonitoring neurotoxic contaminations in tropical environments. Results show that ChE activities in both tissues decreased significantly with increased total length (4-24.5 cm) or body weight (1-186 g) of the fish and the relationships were curvilinear. Comparisons of the slopes and elevations of the regression lines of the logarithmic ChE and body size relationships of males with those of females indicated that gender had no significant effect on the body size-specific ChE activities. Response of the ChE of sexually mature males to chlorpyrifos exposure was similar to that of females. Gonadal maturity stage of this fish does not seem to influence ChE activities. Storage of tissues at -80 degrees C for 28 days had no significant effect on ChE activities in the control fish and the fish exposed to carbofuran. However, a partial reactivation of brain ChE activities was observed in the fish exposed to carbosulfan after 28 days of storage. The results emphasize the importance of consideration of body size of the fish and storage time of the tissues in order to formulate accurate conclusions about the neurotoxic chemical exposure when ChE of the fish is used in biomonitoring programs.

Body size-related differences in the inhibition of brain acetylcholinesterase activity in juvenile Nile tilapia (Oreochromis niloticus) by chlorpyrifos and carbosulfan.

Influence of body size on inhibition of brain acetylcholinesterase (AChE) activity of juvenile Nile tilapia, Oreochromis niloticus by chlorpyrifos and carbosulfan was investigated concerning its potential use in the biomonitoring of anticholinesterase pesticides in tropical water bodies. Three size groups of fish (fry: 3-4 cm, fingerlings: 6-8 cm, sub-adults: 10-12 cm in total length) were exposed to a series of concentrations of chlorpyrifos (0.5-12 microg L(-1)) or carbosulfan (1-10 microg L(-1)), and concentration-response for inhibition and recovery of the AChE enzyme was evaluated in comparison to the controls at different time points, 2, 6, 10, and 14 d. The AChE activities of the control fish followed the order of decreasing activity, fry>fingerlings>sub-adults. AChE activities of the fry were nearly 2-fold higher than that of the sub-adults. Following 48 h of pesticide exposure, the AChE activity of the three size groups of fish decreased significantly in comparison to the respective controls in a concentration-dependent manner. The activity was greatly inhibited in the fry (39-85%) compared to sub-adults (18-47%) exposed to the most of the similar concentrations of the pesticides. Median effective in vivo inhibition concentrations (48 h IC50) of chlorpyrifos for fry, fingerlings, and sub-adult stages were 0.53, 0.75, and 3.86 microg L(-1), respectively, whereas the corresponding values for carbosulfan were 3.37, 7.02, and 8.72 microg L(-1). When fish were maintained in the initial pesticide medium for 14 days, AChE activity restored gradually depending on the initial pesticide exposure concentration and the size group of the fish. Results indicate that brain AChE of Nile tilapia is a promising biomarker for assessment of anticholinesterase pesticide contaminations in water. However, body size of Nile tilapia should be taken into account when using this biomarker in biomonitoring programmes.

Activation of 14C-toluene to covalently binding metabolites by rat liver microsomes.

14C-Toluene was incubated with rat liver microsomes in the presence of an NADPH-generating system and metabolites were concentrated on cyclohexyl cartridges. The metabolites were separated by reverse phase HPLC and identified by comparing the retention time to standards. 14C-Toluene was converted to 14C-benzylalcohol, 14C-cresols, and an unidentified 14C-metabolite. Some of the radioactivity was found to bind covalently to microsomal macromolecules, preferentially to proteins. The binding was proportional to incubation time and microsomal protein concentration and required NADPH and molecular oxygen. The binding was greatly diminished when microsomes were heat denatured. The binding process was partially inhibited by carbon monoxide and SKF 525-A. When microsomes from phenobarbital- and 3-methylcholanthrene-treated rats were employed, binding was enhanced by 8- and 4-fold, respectively. The binding process was effectively diminished by the presence of reduced glutathione or cysteine in the incubation mixture and was not affected by lysine. Styrene oxide greatly enhanced binding. UDP-glucuronic acid, superoxide dismutase, and ascorbic acid also diminished the binding to some degree. It was concluded that toluene undergoes a hepatic microsomal monooxygenase-mediated activation, and the resultant reactive metabolites binds covalently to microsomal proteins.

A comparative study of the effects of benzene, toluene, and xylenes on their in vitro metabolism and drug-metabolizing enzymes in rat liver.

Male Sprague-Dawley rats were injected ip with benzene, toluene, or a mixture of xylene isomers at 20 mmol hydrocarbon/kg daily for 3 days. The effects of administration of these hydrocarbons upon their own in vitro metabolism, as well as upon cytochrome P-450, NADPH-cytochrome c reductase, aminopyrine N-demethylase, aniline hydroxylase, glutathione, glutathione S-transferase, and UDPglucuronyltransferase in liver were studied. Each hydrocarbon studied increased its own in vitro metabolism. Benzene had no effect on the metabolism of toluene or xylenes. Toluene and xylenes increased the metabolism of benzene, toluene, and xylenes. Cytochrome P-450 was elevated by toluene and xylenes, but was not affected by benzene. NADPH-cytochrome c reductase was induced by all three hydrocarbons. Aminopyrine N-demethylase and aniline hydroxylase were induced by toluene and xylenes and were not affected by benzene. Glutathione was elevated by benzene, decreased by xylenes, and not affected by toluene. Glutathione S-transferase was induced differentially by these hydrocarbons toward various substrates: toward 1-chloro-2,4-dinitrobenzene by benzene and toluene, toward 1,2-dichloro-4-nitrobenzene by benzene and xylenes, and no effect toward 1,2-epoxy-3-(p-nitrophenoxy)propane by any hydrocarbons. UDPglucuronyltransferase was induced by benzene and toluene when o-aminophenol and phenol were used as the substrate. Xylenes had no effect. Benzene was more effective at inducing conjugation enzymes. Xylenes were more effective at inducing cytochrome P-450 dependent enzymes. Toluene was equipotent at inducing both types of enzymes. The results indicate that the addition of methyl groups to the aromatic ring affects the inductive pattern of these monocyclic aromatic hydrocarbons.