Antiestrogenicity and estrogenicity in leachates from solid waste deposits

A great deal of effort has been devoted to developing new in vitro and in vivo methods to identify and classify endocrine disrupting chemicals that have been identified in environmental samples. In this study an in vitro test based on recombinant yeast strains transfected with genes for the human estrogen receptor α was adapted to examine the presence of estrogenic and antiestrogenic substances in six Swedish landfill leachates. Antiestrogenic effects were measured as inhibition of the estradiol induced response with the human estrogen receptor α, and quantified by comparison with the corresponding inhibitory effects of a known antiestrogen, hydroxytamoxifen. The estrogenicity was within the range of that determined in domestic sewage effluents, from below the limit of detection to 29 ng estradiol units L^?1. Antiestrogenicity was detected in some of the investigated landfill leachates, ranging between <38 and 3800 μg hydroxytamoxifen equivalents L^?1. There was no apparent relation between the type of waste deposited on the landfills and the antiestrogenic effect. Fractionation of a landfill leachate showed that estrogenic compounds were located in two dominant fractions. Three estrogenic compounds were found that accounted for the estrogenic activity in extracts of leachates: bisphenol A, estradiol, and ethinylestradiol. The bisphenol may have been released from decomposing plastic waste and the estrogenic steroids from earlier deposits of municipal sewage sludge and pharmaceutical waste. Fractionation of leachates from three parts of a landfill showed that the antiestrogenic activity was distributed in at least four fractions and somewhat different in different flows of leachate. This indicated a heterogeneous mixture of antiestrogenic substances. © 2009 Wiley Periodicals, Inc. Environ Toxicol, 2011.     

Determining toxicity of leachates from Florida municipal solid waste landfills using a battery-of-tests approach

The toxicity and physicochemical parameters of municipal solid-waste (MSW) landfill leachates from six sites in north and north-central Florida, United States, were determined. Landfill leachates are complex mixtures of organic and inorganic constituents. Leachate toxicity was assayed using the acute C. dubia (48-h) and Microtoxtrade mark (15-min) assays and the chronic S. capricornutum (96-h) assay. The landfill leachates were shown to be highly toxic to both C. dubia and S. capricornutum with an EC(50) < 10% and < 15%, respectively. However, Microtoxtrade mark was not as sensitive to the leachates. Based on these results, the assays were ranked for their sensitivities to the landfill leachates; C. dubia > S. capricornutum > Microtoxtrade mark. Chemical analyses showed high concentrations of un-ionized ammonia and salts in the landfill leachates but low concentrations of heavy metals. The toxicity of the landfill leachates varied between the sites sampled and within each of the landfill sites. A significant relationship was found between the total ammonia content of the leachates and toxicity as determined by the C. dubia and S. capricornutum assays. Although the chemical constituents in Florida MSW landfill leachates may be more dilute, the toxicity of the leachates is equivalent to that from industrial waste landfills.     

Cytogenetic effects of leachates from tannery solid waste on the somatic cells of Vicia faba

The contamination of surface- and groundwater by the leaching of solid wastes generated by industrial activities as a result of water runoff and rainfall is a matter of great concern. The leachates from tannery solid waste (TSW), a major environmental pollutant, were examined for their possible genotoxic effects on the somatic cells of Vicia faba. Leachates were prepared from solid wastes procured from leather-tanning industrial sites, and V. faba seedlings were exposed to three test concentrations, 2.5%, 5%, and 10%, through soil and aqueous media for 5 days. The root tips examined for cytogenetic damage revealed that leachate of TSW significantly inhibited the mitotic index and induced significantly frequent chromosomal and mitotic aberrations (CA/MA) in a dose-dependent manner. The chemical analysis of TSW samples revealed that the chief constituents were chromium and nickel, which may cause genetic abnormalities. The frequency of aberrations was found to be higher in the root meristematic cells of Vicia faba exposed through the aqueous medium than those exposed through the soil medium. The results of the present study indicated that contamination of potable water bodies by leachates of TSW may cause genotoxicity. For the biomonitoring of complex mixtures of toxicants with the V. faba bioassay, the use of the aqueous medium seems to be a more promising method than the use of the soil medium.     

Genetic damage induced by electronic waste leachates and contaminated underground water in two prokaryotic systems

The inappropriate and unsafe management practices related to disposal and recycling of electronic wastes in Nigeria has led to environmental and underground water contamination. Reports on the level and type of contamination as well as the possible DNA damage effects of this contamination are insufficient. This study evaluated the DNA damaging potential of e-waste simulated and raw leachates, and its contaminated underground water using the SOS chromotest on Escherichia coli PQ37 and the Ames Salmonella fluctuation test on Salmonella typhimurium strains TA98 and TA100, without and with metabolic activation. Physico-chemical parameters of the samples were also analyzed. The result of the Ames test showed induction of base pair substitution and frameshift mutation by the test samples. However, the TA100 was the more responsive strain for the three samples in terms of mutagenic index in the absence and presence of metabolic activation. The SOS chromotest results were in agreement with those of the Ames Salmonella fluctuation test. Nevertheless, the E. coli PQ37 system was slightly more sensitive than the Salmonella assay for detecting genotoxins in the tested samples. Lead, cadmium, manganese, copper, nickel, chromium, arsenic, and zinc contents analyzed in the samples were believed to play a significant role in the observed DNA damage in the microbial assays. The results of this study showed that e-waste simulated and raw leachates, and its contaminated underground water are of potential mutagenic and genotoxic risks to the exposed human populace.     

Chemical composition and toxicity of waste dump leachates using Selenastrum capricornutum Printz (Chlorococcales,Chlorophyta)

Chemical composition and toxicity of leachates from a municipal waste dump in Buenos Aires Province were analyzed. Three sets of samples, obtained in 1996 and 1997, were compared. Levels of nitrogen, phosphorus, and organics (expressed as COD) were distinctly lower in 1997. Such differences were mainly attributed to higher rainfall in that year. The Selenastrum capricornutum assay showed a high sensitivity to heavy metals (Cd=0.049 mg L⁻¹; Cr hexavalent=0.247 mg L⁻¹ and Zn=0.037 mg L⁻¹). Cu, Zn, and Pb concentrations were toxic to algae in leachate samples of the first year, while the EC₅₀ leachates were slightly toxic (86.6 and 99.31%). Most likely, high levels of organics (COD=4640 and 3470 mg L⁻¹) form complex mixtures with metals reducing their toxicity. The EC₅₀ leachate=48.5% in the last year may be explained by high concentrations of Pb and Cu. © 2000 John Wiley & Sons, Inc. Environ Toxicol 15: 76–80, 2000     

Toxicity analysis of leachates from hazardous wastes via microtox and daphnia magna

One of the most common ways of surface and ground water contamination from hazardous wastes is through its leachates. Toxicity is a meaningful parameter allowing an integrated evaluation of the potential danger of leachates. Two bioassays were considered for measuring toxicity: Microtox and Daphnia magna. The toxicity was measured on leachates obtained, using different leaching procedures, from wastes of a pesticide manufacturing industry and sludge from electroplating wastewater. Twenty-two tests were carried out, measuring IC50 for D. magna and EC50 for Microtox. Both bioassays were compared using three toxicity criteria. The first criterium is classification on toxic/non-toxic at 25 and 50% leachate concentration. The second criterium is classification on percent ranks, and the third on log ranks. Considering these criteria, the agreement between both bioassays is within a 75–85%. It is shown that both, Microtox and D. magna assays could be used as toxicity indicators for the wastes considered. The sensitivity of the bioassays is different depending on leachate composition. In all samples, the leachate concentration of chemicals was measured.     

Adverse effect of tannery waste leachates in transgenic Drosophila melanogaster: role of ROS in modulation of Hsp70, oxidative stress and apoptosis

Leachate is a complex chemical mixture of chemicals produced as a result of leaching of solid wastes. The potential toxicity of leachates is a major environmental health concern. The present study evaluated the role of ROS in tannery leachates induced Hsp70 expression, antioxidant enzymes and apoptosis in Drosophila. Different concentrations (0.05-2.0%) of leachates prepared from tannery waste at different pH (7.00, 4.93 and 2.88) were mixed with Drosophila food and fed to the larvae for 2-48 h to examine the different stress and apoptotic markers. A concentration- and time-dependent significant increase in Hsp70 expression, ROS generation, antioxidant enzymes activities and MDA content were observed in the exposed larvae. Activities of antioxidant enzymes were delayed compared with Hsp70 expression and MDA level in the exposed organisms. Apoptotic cell death was observed in the exposed larvae at higher concentrations concurrent with a significant regression in Hsp70 along with a higher level of ROS generation. A positive correlation drawn between ROS generation and apoptotic markers and a negative correlation between apoptotic markers and Hsp70 expression at these concentrations indicated the important role of ROS in the induction of cellular damage in the exposed organisms. There was a significant generation of ROS in the larvae exposed to 0.5% of leachates which did not interfere with the protection of their cells by Hsp70 and antioxidant enzymes. However, generation of significantly higher levels of ROS in the larvae exposed to 1.0% and 2.0% leachates may decrease Hsp70 expression thus leading to mitochondria-mediated caspase-dependent apoptotic cell death.     

Estrogenic effect of leachates and soil extracts from lysimeters spiked with sewage sludge and reference endocrine disrupters

Several experiments were conducted to evaluate the behavior and performance of some potential endocrine disrupters (ECDs). Two in vitro screening assays, one based on MCF7-cell proliferation (E-screen test) and the other on estrogenic receptor activity [enzyme-linked receptor assay (ELRA)], were used for the tests, which were done in lysimeters 80 cm in diameter with depth of 30 cm (shallow) or 90 cm (deep). A sandy soil was used to fill in all lysimeters, which were spiked on the surface with either: (a) a sewage sludge (SS) at a dose equivalent to 20 tons ha-1; (b) a mixture of reference ECDs, comprising 17 alpha- and 17 beta-estradiol (E2), nonylphenol, octylphenol, and bisphenol A at doses 100 times higher than the maximum concentrations respectively found in the applied SS; or (c) a mixture of ECDs and SS. After percolation of the lysimeters with rain and/or artificial water, five leachates were sampled from each lysimeter during a period of 210 days. Immediately after the lysimeter percolation experiments, four and six soil fractions were dissected from, respectively, the 30-cm and 90-cm lysimeters and extracted by water. Both the leachate and soil extract samples were analyzed for their estrogenicity using the assays indicated above. The E-screen assay was highly sensitive only for some leachate and extract samples but gave no response for most leachates and soil extracts. The results of the ELRA assay suggests a significantly higher estrogenicity of leachate samples from shallow lysimeters compared with that of leachates from deep lysimeters. In contrast, the estrogenic effect measured for soil extracts of shallow lysimeters was lower than that measured for soil extracts of deep lysimeters. The results of the E-screen assay suggests the occurrence of a fast mobilization of applied ECDs and a moderate retardation effect of native ECDs contained in applied SS in the sandy soil used in the lysimeters. In lysimeters spiked with a mixture of SS and ECDs, the washing-out effect of ECDs in the first leachate fraction decreased, but the distribution of ECDs in the lysimeters increased. The relatively high estrogenic impact measured for soil water extracts suggests that the ECDs were mostly associated with water-soluble fractions of organic matter and/or water-suspended fractions of the mineral soil matrix. The application of SS to agricultural and forest fields may determine the immobilization of ECDs in soil or their movement to surface and/or groundwater. Therefore, an endocrine risk of exposure exists for the water and soil organisms.     

Assessment of geographical variation in the respiratory toxicity of desert dust particles

Abstract

The health consequences of sand particle inhalation are incompletely understood. This project evaluated the respiratory toxicity of sand particles collected at military bases near Fort Irwin USA, in Iraq (Camp Victory, Taji and Talil), and Khost Afghanistan. Our primary focus was on assessing the role of soluble metals in the respiratory toxicity of the sand particles using in vitro and in vivo methods. Replicating rat type II alveolar cell cultures (RLE-6TN) were exposed to sand extracts or vehicle control in serum-free media for ≤24?h. Cytotoxicity was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and assessment of lactate dehydrogenase leakage. The relative in vitro cytotoxicity of the sand extracts was Taji?≈?Talil?>?Afghanistan?>?Camp Victory?≈?Fort Irwin. We also assessed extracts of Camp Victory, Afghanistan, and Taji sand for acute and delayed pulmonary toxicity in rats following intratracheal administration. Assessments included biochemical analysis of bronchoalveolar lavage fluid (BALF) and lung histopathology. The in vitro cytotoxicity assay results were partially predictive of in vivo responses. The more cytotoxic Taji sand extract induced an acute irritant response in rats following intratracheal administration. Rats given the less cytotoxic Camp Victory sand extract had minimal biochemical or cytological BALF changes whereas rats given either the Afghanistan or Taji sand extracts demonstrated BALF changes that were suggestive of mild lung inflammation. Unexpectedly, we observed similar lung pathology in all extract-exposed rats. The results of our study can be used to prioritize future particle inhalation studies or guide epidemiological study design.     

Comparison of phytotoxicity and mammalian cytotoxicity of nontrichothecene mycotoxins

The phytotoxicity and mammalian cytotoxicity of four nontrichothecene mycotoxins (apicidin, sambutoxin, wortmannin, HC-toxin) were compared. Phytotoxicity was evaluated in terms of electrolyte leakage, growth inhibition, and reduction in chlorophyll content. Based on the parameters evaluated, the relative order of phytotoxicity to duckweed (Lemna pausicostata L.) was wortmannin > HC-toxin > apicidin > sambutoxin. A 48-hr exposure to 10 microM wortmannin, HC-toxin or apicidin caused electrolyte leakage from duckweed. The IC50 values for growth inhibition and chlorophyll reduction for wortmannin, HC-toxin, and apicidin were 0.2 and 2.6 microM, 15.4 and 12.6 microM, and 27.7 and 45.3 microM, respectively. Based on the parameters measured, a 72-hr exposure to 100 microM sambutoxin was not toxic to duckweed. Kudzu (Pueraria lobata L.) leaf disc assays revealed a similar trend in relative toxicities, but higher mycotoxin concentrations were required to elicit phytotoxic effects compared to duckweed. All four mycotoxins were cytotoxic to four mammalian cell cultures. However, in contrast to plants, wortmannin was the least toxic (IC50 = 10 to 20 microM) and sambutoxin exhibited a high level of toxicity (IC50 = 0.5 to 1 microM).     

Use of freshwater algae and duckweeds for phytotoxicity testing

The toxicity of contaminated water of different origins and chemicals [Cr(III), Pb(II), Cu(II), Cd(II), pyrene] were tested using four test species: the alga Selenastrum capricornutum (new name Raphidocelis subcapitata), the duckweed Lemna minor, and the crustaceans Thamnocephalus platyurus and Daphnia magna. On the basis of the results obtained, the sensitivity of plant species and problems concerning the interpretation of the phytotoxicity data are discussed. The data indicated that the sensitivities of crustaceans and plant species both to individual contaminants and to mixtures are unpredictable and that there is no reason to consider plant species less sensitive than animal species. Lemna minor is more sensitive than Selenastrum capricornutum. With colored samples, duckweed is preferable for toxicity testing. To raise the predictive utility of the phytotoxicity data, it is recommended that natural water be used in the test procedure.     

Genotoxicity of hazardous leachates from solid wastes evaluated for environmental impact with the ames test

Fourteen series of toxic wastes from various origins (metallurgy, chemical industries, incinerators, industrial treatment plants, etc.) have been studied for their genotoxic properties with the Salmonella reversion assay. To consider realism of environmental impact, a strategy is proposed to assess genotoxicity of the water-soluble fraction of the wastes. Water-extractable micropollutants were further concentrated by liquid–liquid extraction or lyophilization prior to genotoxicity testing. None of the 14 crude aqueous fractions was shown to be mutagenic. On the contrary, positive responses were registered on the concentrated phases of two solid wastes derived from one chemical industry and from the treatment of organophosphorus wastewaters. Chemical analysis did not reveal the presence of known mutagens in the positive extracts. Interacting effects between micropollutants is thus hypothesized to explain genotoxicity.     

Evaluation of the toxic and genotoxic potential of landfill leachates using bioassays

Landfill leachates are liquid effluents with elevated concentrations of chemical compounds that can cause serious environmental pollution. In the south of the state of Santa Catarina, Brazil, a sanitary landfill was installed that employs a system of anaerobic/facultative lagoons for the treatment of its leachate. The present work examined the toxic and genotoxic potential of untreated and treated landfill leachates using bioassays. The chemical, toxic, genotoxic and mutagenic properties of the untreated leachate and the treated leachate were determined. Examination of the chemical properties showed a marked decrease in parameters after treatment, as well as in toxicity towards all the organisms tested. The results of the comet assay demonstrated that both leachates showed genotoxicity in all of the organisms tested, indicating the persistence of genotoxic substances even after treatment. A significant decrease in micronucleated cells was detected in Geophagus brasiliensis exposed to the treated leachate compared to untreated.     

Negative geotropic effect and phytotoxicity of N-quinolinphthalamic acids and allied substances]

N-Quinolinphthalamic acids and allied substances have been prepared and studied for negative geotropic effect and phytotoxic activity. The substances (Tables I leads to III) prepared by fusion of the required anhydride and amine were tested to evaluate the negative geotropic effect on germinating roots of Lens esculenta Moench s.1. and phytotoxicity in both pre- and post-emergence tests against five representative weed species at a dose of 6 kg/ha. It was found that substances (I), (VII), (VIII) and (IX) showed negative geotropic effect of the same order of magnitude as that of the reference substance (alpha-naphthylphthalamic acid). Substances (VII), (VIII) and (IX) proved highly active in the phytotoxicity tests. The simultaneous appearance of geotropic and pre-emergence phytotoxic activity seems to be connected with the presence of an 8-quinoline residue on the amide nitrogen.