Proteomics study revealed altered proteome of Dichogaster curgensis upon exposure to fly ash.

Fly ash is toxic and its escalating use as a soil amendment and disposal by dumping into environment is receiving alarming attention due to its impact on environment. Proteomics technology is being used for environmental studies since proteins respond rapidly when an organism is exposed to a toxicant, and hence soil engineers such as earthworms are used as model organisms to assess the toxic effects of soil toxicants. This study adopted proteomics technology and profiled proteome of earthworm Dichogaster curgensis that was exposed to fly ash, with main aim to elucidate fly ash effects on cellular and metabolic pathways. The functional classification of identified proteins revealed carbohydrate metabolism (14.36%), genetic information processing (15.02%), folding, sorting and degradation (10.83%), replication and repair (3.95%); environmental information processing (2.19%), signal transduction (9.61%), transport and catabolism (17.27%), energy metabolism (6.69%), etc. in the proteome. Proteomics data and functional assays revealed that the exposure of earthworm to fly ash induced protein synthesis, up-regulation of gluconeogenesis, disturbed energy metabolism, oxidative and cellular stress, and mis-folding of proteins. The regulation of ubiquitination, proteasome and modified alkaline comet assay in earthworm coelomocytes suggested DNA-protein cross link affecting chromatin remodeling and protein folding.

Biomarker responses in the earthworm, Dichogaster curgensis exposed to fly ash polluted soils.

Earthworms are globally accepted as a model organism in terrestrial ecotoxicology for assessment of environmental pollution. This study evaluated and compared effects of fly ash polluted soils collected from two geographically different thermal power plants on biomarker responses in the earthworm, Dichogaster curgensis. To evaluate relationship between distance sampling and biomarker responses in the earthworm D. curgensis, soil samples at 0.5, 1 and 3km from thermal plant were analyzed for physico-chemical properties and metal concentrations. Biochemical alterations, lysosomal membrane stability, genotoxic effects, and histological changes were examined on 1, 7, and 14 d of exposure to fly ash contaminated soils collected from different thermal power plants. The activities of superoxide dismutase (SOD), glutathione peroxidase (GPx), and malondialdehyde (MDA) levels were significantly increased, while glutathione reductase (GR) activity was found to be decreased in treated animals. Catalase (CAT) and glutathione-S- transferase (GST) activities were found to be increased initially up to 7d exposure and further decreased on 14d exposure. D. curgensis exposed to fly ash contaminated soils showed significant lysosomal membrane destabilization and DNA damage. Extensive histopathological changes were observed in the tissues of the body wall and intestinal tract of the exposed D. curgensis along with accumulation of heavy metals. These results demonstrate that soil pollution around thermal power plants has adverse biological effects of on the indicator organism D. curgensis and no correlation was found between distance and extent of biological biochemical responses.

Genotoxicity of cadmium chloride in the marine gastropod Nerita chamaeleon using comet assay and alkaline unwinding assay.

This paper presents an evaluation of the genotoxic effects of cadmium chloride (CdCl2 ) on marine gastropod, Nerita chamaeleon following the technique of comet assay and the DNA alkaline unwinding assay (DAUA). In this study, the extent of DNA damage in gill cells of N. chamaeleon was measured after in vivo exposure to four different concentrations (10, 25, 50, and 75 µg/L) of CdCl2 . In vitro exposure of hydrogen peroxide (H2 O2 ; 1, 10, 25, and 50 µM) of the gill cells showed a significant increase in the percentage tail DNA, Olive tail moment, and tail length (TL). Significant changes in percentage tail DNA by CdCl2 exposure were observed in all exposed groups of snails with respect to those in control. Exposure to 75 µg/L of CdCl2 produced significant decrease in DNA integrity as measured by DAUA at all duration with respect to control. In vivo exposure to different concentrations of CdCl2 (10, 25, 50, and 75 µg/L) to N. chamaeleon showed considerable increase in DNA damage as observed by both alkaline comet assay and the DAUA. The extent of DNA damage in marine gastropods determined by the application of alkaline comet assay and DAUA clearly indicated the genotoxic responses of marine gastropod, N. chamaeleon to a wide range of cadmium concentration in the marine environment.

Simultaneous decolorization of reactive Orange M2R dye and reduction of chromate by Lysinibacillus sp. KMK-A.

Azo dyes constitute the largest and diverse group of dyes, widely used in number of industries that are contributing toward organic and inorganic load of effluent treatment. In the present study, Lysinibacillus sp. KMK-A was able to effectively decolorize Orange M2R dye up to 2000 mg l(-1) (Vmax of 19.6 mg l(-1) h(-1) and Km of 439 mg l(-1)) and reduce Cr(VI) up to 250 mg l(-1) (Vmax of 3.6 mg l(-1) h(-1) and Km 28.3 mg l(-1)). It also has an ability of simultaneous decolorization of Orange M2R dye (200-1000 mg l(-1)) with reduction of Cr(VI) (50-200 mg l(-1)). Significant reduction in total organic carbon content, chemical and biological oxygen demand along with spectroscopic and chromatographic analysis confirmed the biotransformation of Orange M2R. Involvement of enzymes namely azoreductase and chromate reductase was observed during biotransformation. The phyto and geno toxicity studies demonstrated that metabolites of dye degradation were non-toxic. Higher tolerance with simultaneous decolorization and detoxification of azo dyes in presence of Cr(VI) makes Lysinibacillus sp. KMK-A, a potential candidate for eco-friendly remediation of metal contaminated dye effluents.

Effect of fly ash on biochemical responses and DNA damage in earthworm, Dichogaster curgensis.

Fly ash is receiving alarming attention due to its hazardous nature, widespread usage, and the manner of disposal; leading to environmental deterioration. We carried out bio-monitoring and risk assessment of fly ash in earthworms as a model system. Dichogaster curgensis were allowed to grow in presence or absence of fly ash (0-40%, w/w) for 1, 7, and 14 d. The biochemical markers viz. catalase (CAT), superoxide dismutase (SOD), glutathione reductase (GR), glutathione peroxidase (GPx), glutathione S-transferase (GST), and malondialdehyde (MDA) level were measured. The comet and neutral red retention assays were performed on earthworm coelomocytes to assess genetic damages and lysosomal membrane stability. The results revealed increased activities of SOD, GPx, GST, and MDA level in a dose-response manner while GR activity was decreased with increasing concentrations of fly ash. No obvious trend was observed in the CAT activity and fly ash concentration. Lysosomal membrane destabilization was noted in the earthworms exposed to 5% and more fly ash concentration in a dose and time dependent manner. The comet assay demonstrated that the fly ash induced DNA damage and DNA-protein crosslinks in earthworm coelomocytes.