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The sub-lethal exposure (20% and 60%) of LC^sub 50^ of herbicide butachlor of different feed materials of buffalo dung with agro-wastes of different exposure periods was observed in the laboratory. There was time and dose dependent Sub-lethal (20% and 60%) of LC^sub 50^ of different exposure periods were observed after treatment of different combination of buffalo dung with agro-wastes. The prolong duration in clitellum development (days), significant reduction in cocoon production/worm and hatchlings/cocoon as well as survival of hatchlings were observed. The sub lethal effect of herbicide butachlor on the earthworm Eutyphoeus waltoni was both time and dose dependent. The prolong duration of clitellum development was observed in 60% LC^sub 50^ of all exposure periods in buffalo dung. Maximum per cenatage of cocoon production, hatchling/cocoon and survival of hatchling was observed in 20% LC^sub 50^ of all exposure periods in buffalo dung, wheat straw and gram bran combination. There was no mortality observed in control. There was no significant (P< 0.05, t- test) difference between control and 20% of LC^sub 50^ of BD+Ws+Gb of all exposure periods shows that the vermicompost of feed material of this combination was more potential for use in agricultural fields because it enhance the tolerance power of earthworm Eutyphoeus waltoni against herbicide butachlor.The sub-lethal exposure (20% and 60%) of LC^sub 50^ of herbicide butachlor of different feed materials of buffalo dung with agro-wastes of different exposure periods was observed in the laboratory. There was time and dose dependent Sub-lethal (20% and 60%) of LC^sub 50^ of different exposure periods were observed after treatment of different combination of buffalo dung with agro-wastes. The prolong duration in clitellum development (days), significant reduction in cocoon production/worm and hatchlings/cocoon as well as survival of hatchlings were observed. The sub lethal effect of herbicide butachlor on the earthworm Eutyphoeus waltoni was both time and dose dependent. The prolong duration of clitellum development was observed in 60% LC^sub 50^ of all exposure periods in buffalo dung. Maximum per cenatage of cocoon production, hatchling/cocoon and survival of hatchling was observed in 20% LC^sub 50^ of all exposure periods in buffalo dung, wheat straw and gram bran combination. There was no mortality observed in control. There was no significant (P< 0.05, t- test) difference between control and 20% of LC^sub 50^ of BD+Ws+Gb of all exposure periods shows that the vermicompost of feed material of this combination was more potential for use in agricultural fields because it enhance the tolerance power of earthworm Eutyphoeus waltoni against herbicide butachlor.
ABSTRACT:
The sub-lethal exposure (20% and 60%) of LC^sub 50^ of herbicide butachlor of different feed materials of buffalo dung with agro-wastes of different exposure periods was observed in the laboratory. There was time and dose dependent Sub-lethal (20% and 60%) of LC^sub 50^ of different exposure periods were observed after treatment of different combination of buffalo dung with agro-wastes. The prolong duration in clitellum development (days), significant reduction in cocoon production/worm and hatchlings/cocoon as well as survival of hatchlings were observed. The sub lethal effect of herbicide butachlor on the earthworm Eutyphoeus waltoni was both time and dose dependent. The prolong duration of clitellum development was observed in 60% LC^sub 50^ of all exposure periods in buffalo dung. Maximum per cenatage of cocoon production, hatchling/cocoon and survival of hatchling was observed in 20% LC^sub 50^ of all exposure periods in buffalo dung, wheat straw and gram bran combination. There was no mortality observed in control. There was no significant (P< 0.05, t- test) difference between control and 20% of LC^sub 50^ of BD+Ws+Gb of all exposure periods shows that the vermicompost of feed material of this combination was more potential for use in agricultural fields because it enhance the tolerance power of earthworm Eutyphoeus waltoni against herbicide butachlor.
KEYWORDS: Herbicide, Butachlor, buffalo dung, Agro-wastes, Sub lethal exposure, Eutyphoeus waltoni, Reproduction , Development.
INTRODUCTION:
Earthworms are the most important biotic component of soils which play an important role in mineralization and breakdown of organic matter. They are also important contributors to the recycling of carbon and nitrogen in the ecosystem and most suitable bio-indicator organisms for testing chemicals of the soil (Edwards and Lofty 1977; Callahan 1988; Goats and Edwards 1988; Cock et al., 1980; Gobi et al., 2004).Earthworm have been used model animals for the study the effect of agrochemicals on soil fauna. Herbicides manifest low toxicity on earthworms, but indirectly can produce the reduction of the populations by decreasing the organic matter input and weed coverage. Earthworms are also important contributors to the recycling of carbon and nitrogen in the ecosystem. The herbicide 2,4-D has very toxic for growth and reproduction of earthworm Eisenia fetida (Helling et al., 2000; Zhou et al., 2007;Corriela and Moreira 2010).Gobi (2010) reported that the percentage of clitellum development decreased with increasing concentration of butachlor. Use of specific herbicides, fungicides and insecticides in the agricultural field can be highly toxic to earthworm and they will suppress or nearly eliminated earthworm population (Williamson 2000; Van- Gestrel and Van Dis 1988; Ribidoux et al., 1999).
In recent years, through a number of studies for assessing the toxicity of pesticides and herbicides to earthworm mortality, growth and reproduction were carried out. The large amounts of organic wastes are produces during the intensive agricultural. Disposal of these large quantities of animal dung and agro-wastes are serious problems. More over many animal wastes caused serious odor and pollution problems. Eutyphoeus waltoni has been found abundantly in the agricultural fields of eastern U.P. and the sufficient number of Eutyphoeus waltoni is necessary for better recycling of wastes in agricultural fields. (Kumar and Singh, 2014). Butachlor has also been reported to be carcinogenic and can adversely disrupt the reproductive process and affect the thyroid and sex steroid hormones in Zebrafish. (Ou et al., 2000; Chang et al., 2011). The toxicity of 2,4-D against the Eutyphoeus waltoni was higher in sandy soil at 24h to 240h exposure periods.(Singh and Singh, 2015). Earthworms can contribute extensively to soil formation through consumption of dead plant and animal matter, mixing of the particles during digesting, depositing their casts throughout the soil column and improving aeration and drainage of the soil burrowing (Kavitha et al., 2011).
The aim of present study was to investigate the sub lethal effect of LC50 of different exposure periods of herbicide butachlor in different combination of buffalo dung with agro wastes of as feed materials under laboratory conditions.
MATERIAL AND METHODS:
The cultured earthworm Eutyphoeus waltoni were used for the experiment. The buffalo dung, wheat straw and gram bran were collected from different part of Gorakhpur district of U.P. India. Commercially available herbicides butachlor (2-chloro 2,6 diethyl N, butoxymethyl acetanilide) was purchased from Aristo biotech and life science Pvt. Ltd., E-26, G.I.D.C. Manjusar, savli, Distt.Vadodara-391755, Gujrat (India) and used in the experiment. The LC50 different exposure periods of butachlor in different combinations of feed materials on the earthworm Eutyphoeus waltoni was recorded by Singh and Singh (2016).
Measurement of Reproduction and Development of Eutyphoeus Waltoni:
The experiment for study of reproduction and development of earthworm Eutyphoeus waltoni were performed by the method of Chauhan and Singh (2012).The experiment were conducted on cemented earth surface. One kg of different combinations of animal dung, agro wastes in different ratio were kept on 30 x 30x 10 cm in bed form at room temperature in dark. Buffalo dung is used as control. The vermicomposting beds were turned over manually every 24 hours for 10 days in order to eliminated volatile substances. After treatment 20% and 60% of LC50 of butachlor of different combination of feed materials of all exposure periods, ten adult earthworms were incubated in each vermibed for the observation of clitellum development, cocoon production, hatchlings and survival of hatchlings. The clitellum development was observed after one week of earthworms inoculation. After this, cocoons were observed each and every day until cocoon production become ceased (about 13- 14 weeks). After isolation, cocoons were freshly laid inside a petridish containing moist filtered paper at 30±2 °C and 70±5 % RH. Hatching of cocoons were determined after stipulated incubation and number of progeny emerged per cocoon were recorded. The percent survivability of hatchlings was recorded after hatching from cocoons in same treated feed materials.
Statistical Analysis:
Each experiment was replicated at least six times. Data were analyzed by student t- test and analysis of variance (ANOVA) in between different exposure periods and different combinations of feed materials for all parameters. After 13 weeks of experiment, the treated earthworms were transferred into untreated same feed materials of buffalo dung and agro-wastes for the observation of withdrawal of clitellum development, cocoon production/worm, hatchlings and survival of hatchlings.
RESULTS:
In control earthworms clitellum development (days), cocoon production/earthworm, hatchlings/cocoon and survival of hatchlings was 26.29±0.53, 32.39±0.54, 3.79 and 2.68, respectively (Table 1-6). The prolong duration in clitellum development (days), reduction in cocoon production/ earthworm and hatchlings/cocoon as well as survival of hatchlings were observed in treatment of sub lethal exposure of LC50 of butachlor of different combination of feed materials of all exposure periods during the experiment. The treatment of 20% and 60% of 24h LC50 of butachlor of BD+WS+GB caused prolong duration in clitellum development29.28±1.16, 32.10±1.01, and maximum reduction cocoon production/ earthworm 23.11±1.18, 21.97±1.15 and hatchling/ cocoon 82.06% and 65.17% as well as survival of hatchling 75% and 67.91%, respectively (Table 1 and fig 1-6). The time and dose dependent effect were observed after treatment of sub-lethal exposure of (20% and 60%) of LC50 of all exposure periods of different combinations of buffalo dung with agro wastes on the clitellum development, cocoon production and hatchlings of cocoon as well as survival of hatchlings of earthworm Eutyphoeus waltoni (Table 1-6 and fig-1-4). The prolong duration in clitellum development (days) and maximum reduction in cocoon production/worm and hatchlings/cocoon as well as survival of hatchlings was observed in the treatment of 20% of 24h LC50 (0.279mg/kg) of buffalo dung with wheat straw and gram bran (BD+Ws+Gb) 20% of 24h LC50 (0.210 mg\kg) of buffalo dung + wheat straw (BD+Ws) and 20% of 24h LC50 (0.306 mg\kg) of buffalo dung with gram bran (BD+Gb) as well as 20% of 24h LC50 (0.208 mg\kg) of buffalo dung respectively on the clitellum development period, cocoon production/ worm and hatchlings/cocoon as well as survival hatchling of Eutyphoeus waltoni, respectively.
No significant (P<0.05,t-test) difference between control and 20% of LC50 of BD+Ws+Gb was observed. The significant analysis of variance (ANOVA)was observed in clitellum development, cocoon production, hatchling and survival of hatchlings in between different exposure periods and sub lethal exposure of different combinations feed materials expect all treatment of 20% of LC50 of BD+Ws+Gb of all exposure periods. (Table 1-6 and fig 1-4).
After 90 days of experiments the treated earthworm were transferred in to untreated feed materials of same combination of wastes materials. There was significant recovery of clitellum development observed after withdraw of treated earthworm in untreated feed materials of BD (37.85+0.62) BD+Ws (34.69+0.58) BD+Gb (35.25+0.37) BD+Ws+Gb (30.65+0.41). The maximum significant recovery was obtained in 20% of LC50 24h of butachlor in the combination of feed material BD+Ws+Gb.
DISCUSSION:
The sub-lethal exposure of (20% and 60%) of LC50 of butachlor for all exposure periods of different combinations of buffalo dung with agro wastes on the clitellum development, cocoon production and hatchlings of cocoon as well as survival of hatchlings of earthworm Eutyphoeus waltoni were time and dose dependent. Herbicides affect the feeding behavior of earthworms, which was reflected in the weight loss and reproduction capacity (Venter et al., 1988; Bustos-Obregon and Goicochea 2002). Indiscriminate use of pesticides may affect non-target organisms in the soil and can cause serious damage to ecosystem (Reinecke and Reinecke 2007).
Smith et al., (1992) reported that soil animals, especially earthworms, are one of the best bio-indicators of pesticide contamination. The agrochemical concentration is higher in surface layers; earthworm activity is very much reduced in the soil surface layer (Keogh and Whitehead 1975; Cock et al.,1980).Gobi (2010) reported that percentage of clitellum development decreased with increasing concentration of butachlor. Similarly Helling et al., (2000) reported that the fungicide copper oxychloride reduced cocoon production with increased concentration of fungicide in Eisenia fetida.
Brown (1978) reported that some herbicides are directly toxic to earthworms while others have virtually no effects. The earthworms represented a major proportion of total biomass of terrestrial invertebrates up to 80% which play an important role in ingestion of large quantity of decomposed litter, manure and other organic matter and convert it into rich top soil (Sandoval et al., 2001). Earthworms are regarded as a reference compartment to observe soil contaminant bioavailability and are used to evaluate the lethal and sub lethal effects of chemicals contaminants and pollutants (Rida 1997). Eguchi et al., (1995) reported that earthworms are considered not only composting agents but also nature's ploughs, aerators, moisture retainers, crushers, and biological agents. Vermicasting have led to significant increases in the yields of several crops, with significant reductions in pesticides use and almost zero chemical fertilizer imputes (Dash and Senapathi 1986).
The Clitellum development and Cocoon production by Eutyphoeus waltoni was more in BD+Ws+Gb than the other combinations. Loh et al., (2004) reported that biomass gain and cocoon production by Eisenia fetida was more in cattle waste than goat waste. Yasmin and Souza (2007) have reported that pesticides influence the reproduction (cocoon production, a reduced mean and maximum number of hatchlings per cocoon and a longer incubation time) of worms in dose- dependent manner, with greater impact at higher concentration of chemical. Xiao et al., (2006) showed that acetochlor had no long term effect on the reproduction of Eisenia fetida at field dose. Choo and Baker (1998) also found that cocoon production in Aporrectodea trapezoides was inhibited by endosulfan and fenamiphos and methiocarb at normal application rates. Negative impact of pesticides on the earthworm growth and development has reported by various researchers. According to Navarro and Obregon (2004), Malathion also has a direct cytotoxic effect causing coiling of the tail, with increase of metachromasia of the chromatin of the spermatozoa and altering the sperm count.
The combination of buffalo dung with wheat straw and gram bran have maximum biomass, weight and length (Nath et al.,2009; Chauhan and Singh 2012; Kumar and Singh 2013). Nath et al., (2009) reported that the feed material of buffalo dung with gram bran have rich organic nutrients. Simileraly BD+Ws+Gb combination have rich organic content which accelerates the clitellum development and cocoon production, hatching and survival of hatchlings Eutyphoeus waltoni. Neuhauser et al., (1979) observed that the food availability and population density have affected the sexual maturation in earthworms. Kaushal et al., (1995) reported that the different cattle dung have differences in growth rate of earthworm Drawida nepalenesis.
CONCLUSION:
Among all the treatment of sub-lethal exposure of LC50 of different combinations of feed materials, the tertiary combination of buffalo dung with wheat straw and gram bran have more potency to increase the tolerance power of earthworm Eutyphoeus waltoni in agricultural fields.
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Vandana Singh, Keshav Singh*
Vermibiotechnology Laboratory, Department of Zoology,
D.D.U. Gorakhpur University, Gorakhpur, 273 009, UP, India
*Corresponding Author Email: [email protected]
Copyright A&V Publications Oct-Dec 2016