Maheswar Lenka

Monitoring and assessment of mercury pollution in the vicinity of a chloralkali plant. IV. Bioconcentration of mercury in in situ aquatic and terrestrial plants at Ganjam, India

In situ aquatic and terrestrial plants including a few vegetable and crop plants growing in and around a chloralkali plant at Ganjam, India were analyzed for concentrations of root and shoot mercury. The aquatic plants found to bioconcentrate mercury to different degrees included Marsilea spp., Spirodela polyrhiza, Jussiea repens, Paspalum scrobiculatam, Pistia stratiotes, Eichhornia crassipes, Hygrophila schulli, Monochoria hastata and Bacopa monniera. Among wild terrestrial plants Chloris barbata, Cynodon dactylon, Cyperus rotundus and Croton bonplandianum were found growing on heavily contaminated soil containing mercury as high as 557 mg/kg. Analysis of mercury in root and shoot of these plants in relation to the mercury levels in soil indicated a significant correlation between soil and plant mercury with the exception of C. bonplandianum. Furthermore, the tolerance to mercury toxicity was highest with C. barbata followed by C. dactylon and C. rotundus, in that order. The rice plants analyzed from the surrounding agricultural fields did not show any significant levels of bioconcentrated mercury. Of the different vegetables grown in a contaminated kitchen garden with mercury level at 8.91 mg/kg, the two leafy vegetables, namely cabbage (Brassica oleracea) and amaranthus (Amaranthus oleraceous), were found to bioconcentrate mercury at statistically significant levels. The overall study indicates that the mercury pollution is very much localized to the specific sites in the vicinity of the chloralkali plant.

Studies on the Ability of Water Hyacinth (Eichhornia crassipes) to Bioconcentrate and Biomonitor Aquatic Mercury

Water hyacinth (Eichhornia crassipes, Mart solms) plants were employed to assess bioconcentration and genotoxicity of aquatic mercury. Plants were exposed to water contaminated with mercuric chloride (MC) or phenyl mercuric acetate (PMA) at 0.001 to 1.0 mg litre(-1), or mercury contaminated effluent from a chloralkali plant for various periods of 4 t0 96 h. Root samples taken after 4, 8, 12, 24, 48, 72 and 96 h of exposure were analysed for bioconcentration of mercury spectrophotometrically, and the root meristems were fixed in aceto-ethanol for cytological analysis to determine the frequencies of cells with micronuclei (MNC). Ethyl methane sulfonate and tap water served as positive and negative controls, respectively. The results indicated that bioconcentration of mercury in root tissue was both time- and concentration-dependent, providing evidence that water hyacinth is a good absorbant of aquatic mercury. The frequency of root meristematic cells with MNC followed a concentration-response. The findings indicate the potential of water hyacinth plants for in situ monitoring and for mitigation of aquatic mercury pollution.

Monitoring and assessment of mercury pollution in the vicinity of a chloralkali plant. I, Distribution, availability and genotoxicity of sediment mercury in the Rushikulya estuary, India

Industrial effluent discharged into the Rushikulya estuary from a chloralkali plant has been identified as the source of mercury which has polluted the estuary for the past two decades. Sediment samples taken from eight sites representing the effluent channel, effluent discharge point and the course of the estuary, covering a distance of 2 km, were analyzed for total mercury. Phthalate buffer (pH 2–6) was used to study leachability of mercury from sediments as a function of pH. Chemical extraction with 10% HNO3, ammonium acetate-EDTA and 0.1 and 0.05 M CaCl2 was used to assess bioavailability of mercury from the sediments. Further, sediment samples were bioassayed by the Allium micronucleus (MNC) assay. The endpoints measured were root length, root mercury (nioconcentrated mercury) and frequency of root meristematic cells with MNC. The mercury concentrations in sediment samples ranged from 1.6 to 192 mg kg−1. The results indicated that pH was an important factor regulating the availability of mercury; the lower the pH the higher the availability of mercury. Of the different chemical extractants used, 0.05 M CaCl2 extractable mercury showed the highest correlation with bioconcentrated mercury (r2 = 0.91, P ⩽ 0.01), emphasizing the fact that 0.05 M CaCl2 was the best predictor of bioavailable mercury from the sediment. Similarly, the frequency of root meristematic cells with MNC was highly correlated not only with bioconcentrated mercury (r2 = 0.92, P ⩽ 0.01), but also with bioavailable mercury, as evidenced by 0.05 M CaCl2 extractable mercury (r2 = 0.85, P ⩽ 0.01). The Allium MNC assay can assess mercury pollution and takes into account the bioavailability of the metal. The overall assessment indicated that, although localized, the sediment of the Rushikulya estuary is contaminated with high concentrations of mercury with respect to both bioavailability and toxicity.

Monitoring and assessment of mercury pollution in the vicinity of a chloralkali plant. II Plant-availability, tissue-concentration and genotoxicity of mercury from agricultural soil contaminated with solid waste assessed in barley (Hordeum vulgare L.).

Barley (Hordeum vulgare L.) was used to assess plant-availability, tissue-concentration and genotoxicity of mercury from the solid waste deposits of a chloralkali plant. Seeds of H. vulgare, presoaked in distilled water, were allowed to germinate and grow on agricultural soil mixed with solid waste containing 2550+/-339 mg Hg kg(-1) at different proportions (0.75, 1.5, 2.5 and 5%). Plants raised from germinating seeds on uncontaminated agricultural soil served as controls. On day 7, germination counts and seedling heights were recorded. The concentration of mercury in soil, and plant tissue (dry weight) were determined at different stages of plant growth from day 7 till maturity and harvest. The availability of mercury from the soil was assessed by extracting mercury at a range of pH values (2-6) and by chemical methods. The embryonic shoots excised at 36 h of germination were subjected to cytological analysis to determine mitotic index and frequency of mitoses with chromosomal aberrations. The pollen mother cells from anthers of young M1-spikes were analysed to score meiotic aberrations. Subsequently, the pollen fertility and seed set were determined. Furthermore, M2-seedlings were analysed for chlorophyll-deficient mutations. The cytogenetic analysis revealed the effects of mercury on the mitotic and meiotic chromosomes which were significantly correlated with soil-mercury. The bioconcentration of mercury in aerial tissues decreased with the age of the plant. Roots, in particular, were found to concentrate most of the mercury taken up by the plant. At the time of harvest, the bioconcentration of mercury in straw was at a minimum. The accumulation of mercury in grain, which was significant, did not increase with the increase in concentration of mercury in soil but maintained a plateau, indicating a restriction of transport of mercury through the phloem. The unique advantage with the use of Hordeum assay is that, besides assessing the germline toxicity, the assay takes into account the possible contamination of the agricultural food-chain.

Allium micronucleus (MNC) assay to assess bioavailability, bioconcentration and genotoxicity of mercury from solid waste deposits of a chloralkali plant, and antagonism of l-cysteine

Samples of solid waste from a chloralkali plant were bioassayed employing the Allium micronucleus (MNC) assay. The endpoints measured were root length, root mercury and frequency of root meristematic cells with MNC. Chemical extraction methods such as 10% HNO3, ammonium acetate-EDTA and 0.05 or 0.1 M CaCl2 were used to assess the bioavailability of mercury from soil contaminated with solid waste. Analysis of mercury was by cold vapour atomic absorption spectrophotometry. The frequency of MNC induced by contaminated soil, which followed a concentration-response curve, was statistically correlated with soil mercury, extractable mercury and root mercury. The antagonism of L-cysteine against the suppression of root growth and induction of MNC by solid waste suggested the involvement of mercury as well as thiol groups in the process. The threshold assessment values, such as the median effective concentration (EC50) for root growth, gross toxicity concentration tested (GTCT) as indicated by complete inhibition of sprouting of roots, lowest effective concentration tested (LECT) and highest ineffective concentration tested (HICT) for induction of MNC, were determined. These assessment values may be useful for environmental management and regulatory purposes. Furthermore, the detection limit of the Allium MNC assay for solid waste mercury as indicated by LECT was 9.6 mg kg-1 which corresponded to 0.13 mg kg-1 of 0.05 M CaCl2 extractable or 8.3 mg kg-1 dry weight bioconcentrated mercury.

Water hyacinth (Eichhornia crassipes) to biomonitor genotoxicity of low levels of mercury in aquatic environment

The mitotic cell-cycle duration of root meristematic cells of Eichhornia crassipes as determined by the colchicine labelling method was approximately 24 h at 30 +/- 1 degrees C. In one experiment the intact root meristems of E. crassipes were subjected to 1 h acute exposure to water contaminated with maleic hydrazide (MH), 56 ppm, or methyl mercuric chloride (MMCl), 0.1-0.5 ppm, followed by recovery in tap water for 4-48 h. In a second experiment the roots were subjected to 96 h exposure to water contaminated with MH, 56 ppm, or MMCl, 0.0001-0.1 ppm. In both experiments the cytological end-point measured was the frequency of cells with micronuclei (MNC). In the first experiment, while in the MH-exposed root meristems the frequency of MNC was significant at 40 h of recovery, MMCl induced significant MNC at 12, 20, 24, 40, and 40 h of recovery depending on the concentration. In the second experiment both test chemicals induced MNC which was concentration-dependent in case of MMCl. The highest ineffective concentration tested (HICT) and lowest effective concentration tested (LECT) for MMC determined in this experiment were 0.0005 ppm and 0.001 ppm, respectively. The present work provides evidence that E. crassipes could be a promising in situ environmental biomonitoring assay system.

Mercury-tolerance ofChloris barbata Sw. andCyperus rotundus L. isolated from contaminated sites

Chloris barbata Sw. andCyperus rotundus L. from a mercury contaminated site near a chloralkali plant were tested for relative tolerance to Hg by root-elongation method. The above two species from the Hg-contaminated site exhibited high tolerance to Hg compared to the same species from a non-contaminated site. Tolerance to Hg was higher inChloris barbata than inCyperus rotundus.

Monitoring and assessment of mercury pollution in the vicinity of a chloralkali plant. III. Concentration and genotoxicity of mercury in the industrial effluent and contaminated water of Rushikulya estuary, India

Aquatic mercury pollution of the Rushikulya estuary in the vicinity of the chloralkali plant at Ganjam, India was monitored over a period from October 1987 to May 1989. The concentrations of aquatic mercury in the water samples taken from the effluent channel and from different sites along the course of the estuary covering a distance of 2 km were periodically recorded and ranged from 0 to 0.5 mg/l. The bioconcentration and genotoxicity of aquatic mercury in the samples were assessed by the Allium micronucleus (MNC) assay. The frequency of cells with MNC was highly correlated not only with bioconcentrated mercury (root mercury) but also with the levels of aquatic mercury. The threshold assessment values such as effective concentration fifty (EC50) for root growth, lowest effective concentration tested (LECT), and highest ineffective concentration tested (HICT) for induction of MNC in Allium MNC assay for the present aquatic industrial mercury were determined to be 0.14, 0.06 and 0.02 mg/l, respectively.