农药垃圾渗滤液毒性评价和hazard quotient derivation by Allium assay
Sarvesh1*and Prabhakar P. Singh1
1Department of Environmental Sciences, Dr. R.M.L. Avadh University, Faizabad, 2244 001 India
DOI:http://dx.doi.org/10.12944/CWE.2.2.20
本研究设想探索的影响pesticide solid waste leachate to Allium cepa (common onion) bulbs. Bulbs exposed to the leachate showed hampered root growth and morphological deformities. At 15% and higher concentrations of leachate after 5 days, gall like swellings were noticed around the mitotic zone (zone of root growth). From the dose response curve, the EC50 values for the 5th, 10th, 15th and 20th day were calculated and the highest EC50 value, 24.9%, was for the 5th day while the lowest EC50 value, 20%, was for the 20th day. The EC50 decreases from 5th to the 20th day successively. The 5th day EC50 was used to determine the hazard quotient (HQ) of the pesticide waste leachate from the dumping site. A value of 4.49 as HQ suggests that that there is considerable risk, as any value of HQ above one (>1) is environmentally unacceptable. The response in root growth pattern, residue analysis and the HQ indicates that the leachate is toxic to rot growth of onion and steps for proper management of hazardous waste and leachate are urgently required.
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Sarvesh, Singh P.P. Pesticide waste leachate toxicity evaluation and hazard quotient derivation by Allium assay. Curr World Environ 2007;2(2):221-224 DOI:http://dx.doi.org/10.12944/CWE.2.2.20
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Sarvesh, Singh P.P. Pesticide waste leachate toxicity evaluation and hazard quotient derivation by Allium assay. Curr World Environ 2007;2(2):221-224. Available from://www.a-i-l-s-a.com/?p=693
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Article Publishing History
Received: | 2007-11-12 |
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Accepted: | 2007-12-23 |
Introduction
According to a 1997 market estimate, approximately 5684 million pounds of pesticide active ingredients are applied throughout the world1and the release of waste from all pesticide industries will be in direct proportion to the quantity of pesticide produced. Pesticides leaching from agricultural fields and dumping sites to groundwater is an environmentally relevant and highly variable process.2Sub-surface leaching of contaminants from landfills and waste dumps causes severe degradation of the groundwater, at many places, exceeding the WHO permissible limits in drinking water.3Water quality impairment due to leachate contamination can occur due to either percolation downwards or lateral movement according to the slope,4deteriorating the surrounding water bodies. Several monitoring studies have revealed groundwater contamination by pesticides.5
Risk from solid waste dumps/disposals and contaminated soils has been reported in many studies6,7和风险特征的估计incidence and severity of the effects likelihood to occur in an environmental compartment due to actual or predicted exposure to a chemical. The hazard quotient (HQ), ratio of predicted or measured environmental concentrations (PECs) to predicted no-effect concentrations (PNECs), is used for risk characterization.8The present study deals with the residue analysis of the leachate of waste produced by a pesticide industry, engaged in commercial production of lindane (gamma isomer of hexachlorocyclohexane, ã-HCH), its effect on the growth ofAllium ceparoot and derivation of risk quotient associated with the pesticide waste dump.
Material and Methods
Pesticide Waste and Leachate
The pesticide solid waste for the preparation of the leachate was collected from the freshly deposited dumping site of a pesticide industry situated at Chinhut, Lucknow, Uttar Pradesh, India. The samples were brought to the laboratory in airtight bags. The batch leaching test EN12457-2 proposed by the European Commission9(EC), was employed to prepare the leachate for the toxicity testing.
Residue analysis of leachate
Pesticide residue analysis was performed as per methodology of Singhet al.10
Test organism and growth conditions
Bulbs of common onion,A. cepa(diameter 15-20 mm, weight 3-4 gm), were purchased from a certified shop and were properly cleaned, after peeling out the outer scales the bulbs were left under running water for 30 minutes before using for experimental purpose. During the experiment the bulbs were maintained in a BOD incubator at a temperature of 20±2°C in darkness for proper root growth.
Test solutions
The leachate was diluted with tap water to obtain the desired test concentrations of 5.0%, 7.5%, 10%, 15%, 20%, 25%, 50% and 100%. A similar set of control was also run in parallel along with the test concentrations with tap water as growth medium.
Figure 1: Growth of A. cepa root on 5th day, 10th, 15th and 20th day (EC50=24. 25%, 21.49%, 20.6% and 20% respectively) Click here to view figure |
Determination of EC50value
Bulbs were placed over test-tubes filled with different test concentrations of the leachate. For each concentration three bulbs were used. Using no pregrowth of the roots, the experiment will mimic a continuous exposure that is often prevailing in the environment or in daily life. Concentrations were renewed every 24-hour and the length of root bundles, length of ten roots from each bulb, were measured on the 5thday as described by Fiskesjo.11The root lengths were also measured on 10th, 15thand 20thday. The lengths of the root bundles in control bulbs were also measured. Considering control root length as standard, average length of the roots of treated bulbs on the 5th, 10th, 15thand 20thday were plotted against different concentrations and EC50values test were calculated by graphical interpolation.12
Elucidation of the risk quotient
Characterization of risk by the generation of leachate from the waste site is performed by using the EC50value of the leachate forA.cepaexposed to 5 days. PEC was determined by direct monitoring i.e., chemical residue analysis of the leachate13while PNEC was calculated from the lowest EC50value and applying the extrapolation factor, according to Calow and Forbes14and Girling et.al.15The HQ was derived by the use of PEC/PNEC.16
Results
Residue analysis (Table 1) of the pesticide waste leachate shows that it contains 205.50±41, 167.69±1.52, 438.01±6.22 and 87.2±1.13 µg/l of , β, γ and δ hexachlorocyclohexane (HCH) respectively i.e., 0.898 mg/l of HCH (sum of all the isomers).
On the 5thday in 5% concentration, 98.8% growth was recorded while at 100% leachate concentration, root growth was 4.1%. On the 10th
Table 1: Analysis of residues in leachate of pesticide waste collected from the dumping site
S. No. | Hexachlorocyclohexane (HCH) Isomer | Values (µg/l) |
1 | α- HCH | 205.5±4.10 |
2 | β- HCH | 167.69±1.52 |
3 | γ- (HCH | 438.01±6.22 |
4 | δ- HCH) | 87.2±1.13 |
Table shows the values obtained after the analysis of the leachate of the pesticide solid waste; the various isomers of HCH are present in varying amounts and are subsequently used to derive the predicted or measured environmental concentration (PEC). day, the growth in 5% concentration was 99.5% while at 100% concentration growth was 3.9%. On the 15thday the difference between 50% and 100% concentration was maximum (39.94%) and on 20thday this difference was 40%.
Determination of EC50value
EC50values ofA. ceparoot growth were 24.25%, 21.49%, 20.6% and 20% (Fig. 1) on 5th, 10th, 15thand 20thday respectively. Besides control, maximum growth was recorded in 5%, 7.5%, 10% and 15% concentration during 5thto 10thday. However, reduction in growth was observed in these sets during 10thto 15thand 15thto 20thday. In the 100% concentration the roots ceased to grow from 8thday onwards. During the total exposure period the maximum growth (for a 5 day period) of 51.4% was recorded for the 50% concentration during 10thto 15thday. On the 5thday at 25% (H” 5thday EC50) concentration, the roots showed curls and twists.
On the 20thday at 20% (= 20thday EC50) concentration gall like swellings were noticed around the root hair zone.
Elucidation of the hazard quotient
The PEC of 0.898 mg/l was derived from the residue analysis of the leachate while PNEC was calculated, as 0.20 mg/l. Risk from leachate generation in the form of hazard quotient was determined to be 4.49. The residue analysis of the leachate gave the PEC and the EC50was used to derive the PNEC, which were later used to characterize risk.
Discusssion
The EC50values decreased with increase in the exposure duration. A finding by Wardet al.172002 in which landfill leachates were shown to have high toxicity to bothC. dubiaandS. capricornutumwith an EC50< 10% and < 15% respectively suggests that the leachate we tested in this study was moderately toxic. The abnormalities in the growth of roots were in agreement with Fiskesjo,18according to which the growing roots tips may, after various treatments, take the shapes of hooks, spirals or tumours. Such observations may give information of specific action of a chemical.19The growth ofAlliumroot was inhibited by the leachate in a dose-dependent manner and any inhibition of growth reflects toxic effects on metabolic processes.20Results of study of comparative effects of insecticides on theAlliumand mammalian test systems indicate that theAlliumtest responds to these insecticides similar to mammalian test systems21and hence elicits a correlation betweenAlliumroot meristem cells andin vivo/in vitromammalian test systems. These results can be used to relate the leachate toxicity to mammals; since the leachate is toxic toAllium(hampering the root meristem cells) it will be toxic to mammals also.
Similarly obtainment of PEC, PNEC and RCR values was accomplished by Palma,et. al.22,15A PEC/PNEC quotient e” 1 signifies that there is risk of effects occurring and a large quotient is likely to indicate a high level of risk.14Thus a RCR of 4.492 indicates towards moderate to high risk.
Fisksejo11and Chauhan and Sunderaraman23have given consideration for 5thday measurement of root lengths. This 5thday concept may have arisen for the assessment of toxicity of pesticidal sprays and formulations because the pesticides are applied at a particular time and there are intervals between the successive applications. So the impact of pesticide can be evaluated after 5thday from the time of exposure. But in case of leachate toxicity from a solid waste site there is continuous leaching to the nearby soil or water body, therefore, the test was extended to 20 days. A growth period of 48 or 72 hour is suitable as a standard; after 7-8 days the differences between treated and control roots were even clearer, and thus the longer treatment preferred.19The EC50values, for root inhibition, reported here can be used to determine appropriate leachate concentrations for genotoxicity assays.
As a conclusion it is recommended that a proper disposal of solid waste and regular monitoring of ecological effects of the leachates generated from such dumping site is essential for preservation of water quality and ecosystem health.
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