Combined use of ecotoxicity tools and physicochemical analysis for evaluating potential toxicity of treated natural rubber processing effluents and receiving waters.

Potential toxicity of treated effluents of selected natural rubber processing industries was evaluated by integrating physicochemical analysis with Daphnia magna and Poecilia reticulata bioassays as ecotoxicity tools. Further, the efficacy of the constructed wetland treatments practiced by the industries for reducing the ecotoxicity of the final effluents reaching the receiving water course was assessed. Even after passing through the constructed wetlands, some of the measured physicochemical parameters of the final effluents did not comply with the stipulated rubber processing effluent regulatory limits. Acute toxicity data of treated effluents demonstrated greater susceptibility of D. magna compared to P. reticulata. Erythrocytic abnormality tests with P. reticulata revealed that rubber industry effluents contained cytogenotoxic contaminations which had not been completely eliminated by the treatment processes. Wetland treatment technique was not effective in reducing the cytogenotoxic effects of final effluents reaching the receiving water course. The use of ecotoxicity tools for optimization of rubber industry effluent treatment processes would help to reduce potential toxic/cytogenotoxic effects of effluent receiving waterbodies considering sustainable development goals focusing on ecosystem safety.

Comparison of Toxicity of Nano and Bulk Titanium Dioxide on Nile Tilapia (Oreochromis niloticus): Acetylcholinesterase Activity Modulation and DNA Damage.

This study compared effects of low concentrations (0.05 and 0.1 mg/L) of nano-TiO2 and bulk-TiO2 on brain, gill and liver acetylcholinesterase (AChE) and erythrocytic DNA of Nile tilapia over 7 and 14 days exposure. Both TiO2 forms did not affect brain AChE activities. Bulk-TiO2 induced elevation of gill AChE activities only after 7 days while nano-TiO2 had no effect. Liver AChE activities were increased by 0.1 mg/L bulk- and nano-TiO2 to similar extents. At 7 days, erythrocytic DNA damage was induced only by 0.1 mg/L nano- and bulk-TiO2 to similar extents, but damage was not repaired to control levels at 7 days recovery period. At 14 days continuous exposure, DNA damage was induced by 0.05, 0.1 mg/L nano-TiO2 and 0.1 mg/L of bulk-TiO2 to similar extents. Results show that both forms of TiO2 can pose genotoxic hazards to fish populations under sub-chronic exposure. However, their neurotoxic potential was not evident.

Concentrations of trace metals in Siganus javus captured in Negombo estuary, Sri Lanka: Human health risk assessment through dietary exposure.

Concentrations of aluminium, cadmium, chromium, lead, nickel and zinc in muscle and liver of a marine fish in Indo-Pacific region, Siganus javus captured in Negombo estuary were evaluated and potential human health risks associated with dietary exposure were assessed. Of the six metals analyzed, zinc was the most abundant metal in both tissues. No significant differences were found between muscle and liver with respect to cadmium and lead concentrations whereas concentrations of other metals were higher in liver compared to muscle. In human health perspective, estimated target hazard quotients for cadmium and lead were greater than the threshold of one indicating potential non-cancer health risks to heavy consumers. Estimated excess cancer risk of cadmium indicate carcinogenic health risks associated with their consumption even at moderate meal frequencies. The results revealed that consumption of S. javus from the estuary needs to be limited concerning non-cancer and cancer risks to human health.

Toxicity of Titanium Dioxide Nanoparticles to Tadpoles of Asian Common Toad (Duttaphrynus melanostictus) Following Short Term and Chronic Exposures.

Nanotoxicity data for amphibians are limited compared to other taxonomic groups. The present study assessed toxicity of titanium dioxide nanoparticles (nanoTiO2, anatase form, particle size < 25 nm) on tadpoles of Duttaphrynus melanostictus after short term and chronic exposures. Exposure to nanoTiO2 (≤ 100 mg/L) for 96 h had no significant effect on survival but upon transfer to nanoTiO2 free water for another 21 days, survival of the tadpoles pre-exposed to nanoTiO2 was greatly reduced and their development was delayed. Chronic exposure to nanoTiO2 (0.1 to 10 mg/L) for 14 days had no significant effects on acetylcholinesterase activities but induced mortalities (up to 40%) and histological alterations in the intestine and liver tissues. The results indicate that release of nanoTiO2 to aquatic ecosystems could pose negative impacts to amphibian populations.

Sensitivity of freshwater organisms to cadmium and copper at tropical temperature exposures: Derivation of tropical freshwater ecotoxicity thresholds using species sensitivity distribution analysis.

Tropical freshwater ecosystems are increasingly influenced by chemical stressors including heavy metals posing threats to biodiversity. Adequate ecotoxicity data are not available for native tropical freshwater species for deriving water quality guidelines and for conducting ecological risk assessments. Objectives of this study were (i) to generate freshwater ecotoxicity data for cadmium (Cd) and copper (Cu) for tropical temperature exposures using standard laboratory bioassays with selected freshwater species and (ii) to derive ecotoxicity thresholds (protection concentrations, PC) for tropical freshwater life based on 'tropical temperature-specific exposure' ecotoxicity data. Estimated final chronic toxicity values of the six species tested in the study indicate that the most sensitive species was the crustacean, Moina macrocopa for both metals while the algae Chlorella vulgaris and the plant Lemna perpusilla showed highest tolerance to Cd and Cu respectively. Tropical temperature-specific exposure (25-30 °C) was used as the decision criterion for deriving ecotoxicity thresholds of Cd and Cu for protection of tropical freshwater life based on species sensitivity distribution analysis of the final chronic toxicity data sets which included published toxicity data of selected species in addition to the six species tested in this study. The derived PC99, PC95, PC90 and PC80 values for protection of tropical freshwater life under chronic exposure are 0.5, 1.2, 1.9 and 3.5 µg/L for Cd and 0.34, 0.84, 1.4 and 2.6 µg/L for Cu respectively. These derived threshold chronic values (PC99 and PC95) indicate that the established freshwater quality guidelines based on temperate species for Cu may not provide sufficient protection of the freshwater species in the tropics while the available freshwater guidelines for Cd would provide adequate protection for the tropical freshwater species. The tropical freshwater ecotoxicity thresholds derived in this study may be used with some caution as reference points for site specific ecological risk assessments in the tropics.

Application of cytogenetic model Allium cepa for screening potential cytogenotoxicity of herbal-based hair dyes.

Plant models may be useful as test organisms for initial screening of potential toxicity of personal care products. The objective of the present study was to assess the efficacy of the Allium cepa (common onion) test system as a bioanalytical tool for screening potential cytotoxicity and genotoxicity of herbal-based hair dye formulations. Exposure of black hair dye formulations for 48 hours resulted in root growth retardation and mitosis suppression in the root meristems of A. cepa bulbs indicating concentration dependent cytotoxicity. At the 72 hour post exposure, cytotoxic effects on the roots were reduced but not recovered completely signifying prolong toxic action of the hair dyes. The condensed nuclei was the most frequent nuclear abnormality found in the dye exposed root meristematic cells indicating the cell death process. Induction of micronuclei and chromosomal aberrations in the root meristematic cells even at the post exposure stage indicates persistent genotoxicity of the hair dyes which may be attributed to the interactive effects of chemical mixtures present in the commercial hair dye formulations. The results revealed that A. cepa test system is an effective bioanalytical tool for screening cytogenotoxicity of commercial hair dye formulations.

Cytogenotoxicity screening of source water, wastewater and treated water of drinking water treatment plants using two in vivo test systems: Allium cepa root based and Nile tilapia erythrocyte based tests.

Biological effect directed in vivo tests with model organisms are useful in assessing potential health risks associated with chemical contaminations in surface waters. This study examined the applicability of two in vivo test systems viz. plant, Allium cepa root based tests and fish, Oreochromis niloticus erythrocyte based tests for screening cytogenotoxic potential of raw source water, water treatment waste (effluents) and treated water of drinking water treatment plants (DWTPs) using two DWTPs associated with a major river in Sri Lanka. Measured physico-chemical parameters of the raw water, effluents and treated water samples complied with the respective Sri Lankan standards. In the in vivo tests, raw water induced statistically significant root growth retardation, mitodepression and chromosomal abnormalities in the root meristem of the plant and micronuclei/nuclear buds evolution and genetic damage (as reflected by comet scores) in the erythrocytes of the fish compared to the aged tap water controls signifying greater genotoxicity of the source water especially in the dry period. The effluents provoked relatively high cytogenotoxic effects on both test systems but the toxicity in most cases was considerably reduced to the raw water level with the effluent dilution (1:8). In vivo tests indicated reduction of cytogenotoxic potential in the tested drinking water samples. The results support the potential applications of practically feasible in vivo biological test systems such as A. cepa root based tests and the fish erythrocyte based tests as complementary tools for screening cytogenotoxicity potential of the source water and water treatment waste reaching downstream of aquatic ecosystems and for evaluating cytogenotoxicity eliminating efficacy of the DWTPs in different seasons in view of human and ecological safety.

Combination of physico-chemical analysis, Allium cepa test system and Oreochromis niloticus erythrocyte based comet assay/nuclear abnormalities tests for cyto-genotoxicity assessments of treated effluents discharged from textile industries.

Bioassays for cyto-genotoxicity assessments are generally not required in current textile industry effluent discharge management regulations. The present study applied in vivo plant and fish based toxicity tests viz. Allium cepa test system and Oreochromis niloticus erythrocyte based comet assay and nuclear abnormalities tests in combination with physico-chemical analysis for assessing potential cytotoxic/genotoxic impacts of treated textile industry effluents reaching a major river (Kelani River) in Sri Lanka. Of the treated effluents tested from two textile industries, color in the Textile industry 1 effluents occasionally and color, biochemical oxygen demand and chemical oxygen demand in the Textile industry 2 effluents frequently exceeded the specified Sri Lankan tolerance limits for discharge of industrial effluents into inland surface waters. Exposure of A. cepa bulbs to 100% and 12.5% treated effluents from both industries resulted in statistically significant root growth retardation, mito-depression, and induction of chromosomal abnormalities in root meristematic cells in comparison to the dilution water in all cases demonstrating cyto-genotoxicity associated with the treated effluents. Exposure of O. niloticus to the 100% and 12.5% effluents, resulted in erythrocytic genetic damage as shown by elevated total comet scores and induction of nuclear abnormalities confirming the genotoxicity of the treated effluents even with 1:8 dilution. The results provide strong scientific evidence for the crucial necessity of incorporating cyto-genotoxicity impact assessment tools in textile industry effluent management regulations considering human health and ecological health of the receiving water course under chronic exposure.

Using species sensitivity distribution approach to assess the risks of commonly detected agricultural pesticides to Australia's tropical freshwater ecosystems.

To assess the potential impacts of agricultural pesticides on tropical freshwater ecosystems, the present study developed temperature-specific, freshwater species protection concentrations (i.e., ecotoxicity threshold values) for 8 pesticides commonly detected in Australia's tropical freshwaters. Because relevant toxicity data for native tropical freshwater species to assess the ecological risks were mostly absent, scientifically robust toxicity data obtained at ≥20 °C were used for ecologically relevant taxonomic groups representing primary producers and consumers. Species sensitivity distribution (SSD) curves were subsequently generated for predicted chronic exposure using Burrlioz 2.0 software with mixed chronic and converted acute data relevant to exposure conditions at ≥20 °C. Ecotoxicity threshold values for tropical freshwater ecosystem protection were generated for ametryn, atrazine, diuron, metolachlor, and imidacloprid (all moderate reliability), as well as simazine, hexazinone, and tebuthiuron (all low reliability). Using these SSD curves, the retrospective risk assessments for recently reported pesticide concentrations highlight that the herbicides ametryn, atrazine, and diuron are of major concern for ecological health in Australia's tropical freshwater ecosystems. The insecticide imidacloprid also appears to pose an emerging threat to the most sensitive species in tropical freshwater ecosystems. The exposed temperature-specific approach may be applied to develop water quality guideline values for other environmental contaminants detected in tropical freshwater ecosystems until reliable and relevant toxicity data are generated using representative native species.

Efficacy of Allium cepa test system for screening cytotoxicity and genotoxicity of industrial effluents originated from different industrial activities.

Efficacy of Allium cepa test system for screening cytotoxicity and genotoxicity of treated effluents originated from four types of industrial activities (two textile industries, three rubber based industries, two common treatment plants of industrial zones, and two water treatment plants) was assessed. Physico-chemical parameters including the heavy metal/metalloid levels of the effluents varied depending on the industry profile, but most of the measured parameters in the effluents were within the specified tolerance limits of Sri Lankan environmental regulations for discharge of industrial effluents into inland surface waters. In the A. cepa test system, the undiluted effluents induced statistically significant root growth retardation, mitosis depression, and chromosomal aberrations in root meristematic cells in most cases in comparison to the dilution water and upstream water signifying effluent induced cytotoxicity and genotoxicity. Ethyl methane sulphonate (a mutagen, positive control) and all the effluents under 1:8 dilution significantly induced total chromosomal aberrations in root meristematic cells in comparison to the dilution water and upstream water indicating inadequacy of expected 1:8 dilutions in the receiving waters for curtailing genotoxic impacts. The results support the use of a practically feasible A. cepa test system for rapid screening of cytotoxicity and genotoxicity of diverse industrial effluents discharging into inland surface waters.

Assessing toxicity of copper, cadmium and chromium levels relevant to discharge limits of industrial effluents into inland surface waters using common onion, Allium cepa bioassay.

Toxicity of copper, cadmium and chromium relevant to established tolerance limits for the discharge of industrial effluents into inland surface waters was evaluated by Allium cepa bioassay. The roots of A. cepa bulbs exposed to Cu(2+) (3 mg L(-1)) individually or in mixtures with Cd(2+) (0.1 mg L(-1)) or/and Cr(6+) (0.1 mg L(-1)) exhibited the highest growth inhibition, mitotic index depression and nuclear abnormalities. Root tip cells exposed to Cr(6+) or Cd(2+) alone or in mixture displayed significant chromosomal aberrations in comparison to the controls. EC50s for root growth inhibition followed the order Cu(2+) < Cd(2+) < Cr(6+) indicating greater toxicity of copper. The results show that the industrial effluent discharge regulatory limits for these metals need to be reviewed considering potential cyto-genotoxicity to biological systems.

Chlorpyrifos causes decreased organic matter decomposition by suppressing earthworm and termite communities in tropical soil.

Effects of pesticides on structural and functional properties of ecosystems are rarely studied under tropical conditions. In this study litterbag and earthworm field tests were performed simultaneously at the same tropical field site sprayed with chlorpyrifos (CPF). The recommended dose of CPF (0.6 kg a.i. ha(-1)) and two higher doses (4.4-8.8 kg a.i. ha(-1)) significantly decreased litter decomposition during the first 3 months after application, which could be explained from lower earthworm and termite abundances during this period. Species-specific effects of CPF on organism abundance and biomass were observed, with termites being mostly affected followed by the earthworm Perionyx excavatus; the earthworm Megascolex sp. was least affected. Recovery was completed within 6 months. Decomposition in the controls and lowest two treatments was completed within 4 months, which suggests the need for modification of standard test guidelines to comply with faster litter degradation under tropical conditions.

Assessment of biological effects of pollutants in a hyper eutrophic tropical water body, Lake Beira, Sri Lanka using multiple biomarker responses of resident fish, Nile tilapia (Oreochromis niloticus).

Biomarkers measured at the molecular and cellular level in fish have been proposed as sensitive "early warning" tools for biological effect measurements in environmental quality assessments. Lake Beira is a hypertrophic urban water body with a complex mixture of pollutants including polycyclic aromatic hydrocarbons (PAHs) and Microcystins. In this study, a suite of biomarker responses viz. biliary fluorescent aromatic compounds (FACs), hepatic ethoxyresorufin O-deethylase (EROD) and glutathione S-transferase (GST), brain and muscle cholinesterases (ChE), serum sorbitol dehydrogenase (SDH), and liver histology of Oreochromis niloticus, the dominant fish inhabiting this tropical Lake were evaluated to assess the pollution exposure and biological effects. Some fish sampled in the dry periods demonstrated prominent structural abnormalities in the liver and concomitant increase in serum SDH and reduction in hepatic GST activities in comparison to the control fish and the fish sampled in the rainy periods. The resident fish with apparently normal liver demonstrated induction of hepatic EROD and GST activities and increase in biliary FACs irrespective of the sampling period indicating bioavailability of PAHs. Muscle ChE activities of the resident fish were depressed significantly indicating exposure to anticholinesterase substances. The results revealed that fish populations residing in this Lake is under threat due to the pollution stress. Hepatic abnormalities in the fish may be mainly associated with the pollution stress due to recurrent exposure to PAHs and toxigenic Microcystis blooms in the Lake.

Modulation of ethoxyresorufin O-deethylase and glutathione S-transferase activities in Nile tilapia (Oreochromis niloticus) by polycyclic aromatic hydrocarbons containing two to four rings: implications in biomonitoring aquatic pollution.

Despite ubiquity of polycyclic aromatic hydrocarbons (PAHs) in the tropical environments, little information is available concerning responses of tropical fish to PAHs and associated toxicity. In the present study, effects of five PAHs containing two to four aromatic rings on hepatic CYP1A dependent ethoxyresorufin O-deethylase (EROD), glutathione S-transferase (GST) and serum sorbitol dehydrogenase (SDH) activities in Nile tilapia, a potential fish species for biomonitoring pollution in tropical waters, were evaluated. Results showed that EROD activities were induced by the PAHs containing four aromatic rings (pyrene and chrysene) in a dose dependent manner. However PAHs with two to three aromatic rings (naphthalene, phenanthrene and fluoranthene) caused no effect or inhibition of EROD activities depending on the dose and the duration. Fluoranthene was the most potent inhibitor. SDH results demonstrated that high doses of fluoranthene induced hepatic damage. GST activity was induced by the lowest dose of phenanthrene, fluoranthene and chrysene but high doses had no effect. The results indicate that induction of EROD enzyme in Nile tilapia is a useful biomarker of exposure to PAHs such as pyrene and chrysene. However EROD inhibiting PAHs such as fluoranthene in the natural environment may modulate the EROD inducing potential of other PAHs thereby influencing PAH exposure assessments.

Influence of temperature and soil type on the toxicity of three pesticides to Eisenia andrei.

Expansion of agriculture in the tropics has increased the use of pesticides that may affect the soil ecosystems. Few studies so far determined the effects of pesticides in the tropics and tropical risk assessment therefore often relies on data from temperate conditions. Hence we compared the toxicity of chlorpyrifos, carbofuran and carbendazim to the earthworm Eisenia andrei at two different temperatures reflecting temperate and tropical conditions. The toxicity of the three pesticides in both conditions decreased in the order carbendazim>carbofuran>chlorpyrifos. For chlorpyrifos and carbofuran, but not for carbendazim, survival was more sensitive at the higher temperature, probably due to increased earthworm activity. Sub-lethal effects (reproduction and growth) however, varied inconsistently with temperature and soil types. We conclude that toxicity of pesticides in tropics may not be predicted from data generated under temperate conditions, even within the same species.

In vitro and in vivo effects of cadmium on cholinesterases in Nile tilapia fingerlings: implications for biomonitoring aquatic pollution.

Effects of cadmium on in vitro and in vivo cholinesterase (ChE) activities of brain and muscle tissues of Oreochromis niloticus fingerlings were evaluated, considering its potential use in biomonitoring tropical water pollution. Results show that in vitro ChE activities were depressed significantly by millimolar concentration ranges of Cd(2+). The IC50 values of Cd(2+) on in vitro ChE activity in brain and muscle tissues were 1.56 and 4.31 mM, respectively. Exposure of fish to environmentally relevant concentrations of Cd(2+) (5-30 microg l(-1)) for 28 days evoked only a transient inhibition (21-34%) of in vivo ChE activities. Prior exposure and co-exposure of fish to 15 microg l(-1) of Cd(2+) enhanced the extent of inhibition of ChE levels induced by the organophosphorous insecticide chlorpyrifos. As high concentrations of cadmium have the potential to depress ChE activities, monitoring of metal levels in water bodies with suspected high levels of metal inputs is necessary to accurately interpret the fish ChE inhibition data in relation to insecticide contaminations.

Effect of repeated application of fenthion as a mosquito larvicide on Nile tilapia (Oreochromis niloticus) inhabiting selected water canals in Sri Lanka.

Health status of feral Nile tilapia following repeated applications of fenthion as a mosquito larvicide to selected water canals in Sri Lanka was assessed. With three spray applications of fenthion to the study sites at weekly intervals at the concentration recommended for mosquito control, condition factor and brain acetylcholinesterase activity of the fish were depressed in a time dependent manner. Prominent histopathological alterations displayed were gill hyperplasia and telangiectasis and vacoulation of hepatocytes. Observed ill health effects of fenthion on the fish demonstrate probable ecological risk to the fish populations inhabiting the water canals which receive repeated inputs of fenthion.

Acetylcholinesterase inhibition and gill lesions in Rasbora caverii, an indigenous fish inhabiting rice field associated waterbodies in Sri Lanka.

The present study was aimed at applying condition factor (CF), brain acetylcholinesterase (AChE) and gill histology as biomarkers for detecting possible exposure/effect induced by pesticides in fish residing rice field associated waterbodies in Sri Lanka. Biomarkers of an indigenous fish, Rasbora caverii collected from five sampling sites including canals near rice fields, a river and a reservoir (the reference site) were evaluated at four sampling stages covering pesticide application periods during rice cultivation season in 2004. Results indicated that CF of the fish did not show significant alterations regardless of the sampling sites or sampling stages. Site specific differences in AChE activities of the fish were not evident either prior to application of pesticides or at 7 days after Paraquat application to the rice fields. Two days after the application of a mixture of Fenthion and Phenthoate to the rice fields, AChE activity of the fish collected from canals near rice fields was significantly depressed (65-75%) compared to the fish in the reference site. The activities remain depressed to 50-56% even at 65 days after the insecticides application. Laboratory studies showed that prior exposure of R. caverii to Paraquat (2 microg l(-1), 7 days) enhanced the extent of inhibition of brain AChE activity induced by Fenthion (3 microg l(-1)) or a mixture of Fenthion (3 microg l(-1)) and Phenthoate (5 microg l(-1)). Gills of fish collected from canals near rice fields exhibited abnormal multiple divisions at the tips of some secondary lamellae in addition to hyperplasia, hypertrophy and club shaped deformities. Results indicate that application of pesticides in rice culture could manifest a threat to native fish populations residing rice field associated waterbodies. The response of brain AChE and histological changes in the gills of R. caverii allowed differentiating sampling sites after insecticide applications to the rice fields. Hence, R. caverii may be considered as a surrogate species in ecotoxicological risk evaluation of agrochemicals in the region.