Possible Toxicological Effect of Pesticides , Fipronil On Earthworm ( Eudrilus eugeniae )

Pollution is the preface of infect into an environment, may also defined as an unwanted modify in the physical, chemical and biological characteristics of air, water and soil which influence human life, lives of other useful alive plants and animals, industrial progress, alive conditions and florescent property Water is the most necessary & major necessaries of life. One can rarely survive without water also for a few days. At that 2/3 of the world’s float up region is enclosed by water but yet we are blank. An incubation study was conducted to know the toxicity level of Fipronil in soil fauna by using earthworm species Eudrilus eugeniae as a biomarker. The earthworm were incubated in the spiked soil, with concentration of 17ml, 18ml, 19ml, 20ml for a period of 24hours and 48 hours. From the study, 50% mortality was observed in 17ml. The LC50 concentration were calculated from that it was clear that Fipronil is toxic to earthworm.


Introduction
Soil can also be clear as a natural body which is creation at the border between lithosphere and biosphere by inter chains of the entire ingredient involve in soil arrangement considering both alive and dead. So soil contains not only metal but organic (human) and Organo metal (complex or chilate) combination. Ecological pollution is organism the burning dare of existing living being on the earth. Pollution is the preface of infect into an environment, may also defined as an unwanted modify in the physical, chemical and biological characteristics of air, water and soil which influence human life, lives of other useful alive plants and animals, industrial progress, alive conditions and florescent property. Most of the surface flows of rivers in our nation take place during the heavy rain season of 4 to 5 months and mainly as flood flows. Water toxic waste become when unnecessary equipment go in to water, modification the verity of water (Alrumman et al., 2016) and damaging to atmosphere and human health (Briggs, 2003). Water is a significant natural source used for consumption and other developmental aim in our life (Bidi et al., 2016). An earthworm is a tube-shaped, segmented worm found in the phylum is Annelida. Earthworms are commonly found in living in soil, feeding on live and death organic matter. An earthworm's digestive system runs through the length of its body. It conducts respiration through its skin. I have a double transport system composed of coelomic fluid that moves within the fluid filled coelom and a simple, closed blood circulatory system. It has a central and a peripheral nervous system.
They change soil structure, fertility, develop plant growth and are significant in sustaining productivity (Bucker field, 1998). Earthworm is one which has always interested to mankind; it is nature's own farmer, aerator, crusher, composter, and humidity builder of the top soil and above all is soil's intimate friend and benefactor (Watanabe, 1975 andLal, 1988). The more Chemical cluster of germicides that are generally useful Organophosphate, Carbamate, Chlorinated Hydrocarbons, Pyrethroids and Nicotinoids. Dangerous spraying and shocking management of the chemical pesticides may due to very danger of the health hazard. Fipronil, an Nphenylpyrazole, was introduced into the United States in 1996 for use in animal health, inside pest manage, and commercial turf and crop defense. The use of chemical pesticides has taken place seriously significant to assure both quality and productivity of agricultural products (Oerke and Dehne., 2004;Hayasaka et al., 2012) and to manage household pests such as termites, fire ants, and mosquitoes (Elliott and Barnes, 1963;Drees, 2014). Pesticide coverage is linked with diverse illness along cancer, hormone disruption, asthma, allergies, and hypersensitivity (Van Maele-Fabry et al., 2010). A stroke of proof as well breathe for the opposite effect of pesticide coverage major to birth demerit, abridged birth heaviness, fetal death, etc. (Baldi et al., 2010;Meenakshi et al., 2012;Wickerham et al., 2012).
Pesticides have the possible to due to refuse in farmland bird populations by deplete their food stuff provide. This may occur due to organisms in use as food stuff by farmland birds are either invertebrate vermin or harmful grass that are intentionally notice by pesticides, or because non target organisms that are even diet items for birds are eject occasionally by pesticide application. As these treating do not absorb through poisoning of birds because the toxicity of the energetic ingredients in pesticides or their metabolites, they have be termed not direct impact (Newton, 1995). The analysis that not direct impact of pesticides were drivers of the deny in Grey Partridge populations in Britain was only feasible due to Possible Toxicological Effect of Pesticides, Fipronil On Earthworm (Eudrilus eugeniae) 106 the demography and behavioral biology of the Grey Partridge had been the topic of a compact and continuing study by the Game sanitation faith and others (Potts, 1986;Aebischer & Ewald, 2004).
When croplands are treating, some effects of pesticides take place on non-target terrestrial and marine ecosystem, as well as on adjacent agro ecosystems . As pesticides humiliate, they may filter into soil and water, or they may be desolate or unstable arrive adjacent, or far, areas (Kreisler and Heiss, 2008). Women who consume lots of fish from impure waters have been exposed to have children with high charge of mental, evolution, and behavioral disarray (Ram, 2002). The impact on fish is fall mostly duration the reproductive cycle (Burdick et al., 1967). Some farmers account under estimated the toxicity of the pesticides and thought that throw of report pesticides in runny water was their protected removal. The unless selection to removal of the conclude pesticides was to put in the ground them in fields (Khooharo, 2008). The moiety life of fipronil is 122-128 days in aerobic soils. Under aerobic situation, obviously occurring soil organisms collapse fipronil to form fipronil sulfone. Fipronil can also be hydrolyzed to form fipronil-amide (Bobe et al., 1998).

Fipronil
Fipronil has display neurotoxicity in the severe and sub chronic rat neurotoxicity studies, as well as in the rat chronic/on cogenicity and chronic dog studies. Behavioral changes and loss plenary brain weights were look only at orbit where there was motherly toxicity (decreased body weight, body-weight gain and food consumption) (USEPA, 1998).

Proposed Study
As per statistic, production and productivity have increased. However, the high chemical usage of pesticides to bring about these spectacular increases in food production is not without its problems. A visible parallel correlation between higher productivity, high chemical input use and environment degradation effects is evident in south Gujarat were commercial agriculture is widespread. Crop like rice, maize, jowar, cotton, mangoes, cheeku, bajra, wheat, tobacco, ground nuts, tur, gram all types of vegetables are grown in north Gujarat to get more production farmers use various types of pesticides. Fipronil is most commonly used pesticides which are used as insecticides. Therefore, present work is aimed to evaluate the toxicity effects of widely used pesticides, fipronil an economically important animal Eudrilus eugeniae. An earthworm is a tube-shaped, segmented worm found in the phylum is Annelida. Earthworms are commonly found in living in soil, feeding on live and death organic matter. An earthworm's digestive system runs through the length of its body. It conducts respiration through its skin. I have a double transport system composed of coelomic fluid that moves within the fluid filled coelom and a simple, closed blood circulatory system. It has a central and a peripheral nervous system.
The earthworms are a group of invertebrates belonging to the phylum Annelida and Class Oligochaeta and represented by more than 1000 species. Earthworm is a face organism and it is present in moist and dark places in mud. Earthworms are of great economic value to mankind because they improve the soil quality by their action. Earthworms ingest organic material and facilitate the redistribution of crop residues and organic matter throughout the soil profile. In the Indian subcontinent earthworms are represented by 509 species in 67 genera under 10 families. They are segmented worms, bilaterally symmetrical, with an external gland (clitellum) for producing the egg case (cocoon), a sensory lobe in front of the mouth (prostomium), and an anus at the end of the animal body, with a small number of bristles (setae) on each segment.
Earthworms are a group of soil macro fauna well known for their remarkable contribution in organic matter recycling. The worms have long been related with creative soils. They change soil structure, fertility, develop plant growth and are significant in sustaining productivity (Bucker field, 1998). Earthworm is one which has always interested to mankind; it is nature's own farmer, aerator, crusher, composter, and humidity builder of the top soil and above all is soil's intimate friend and benefactor (Watanabe, 1975 andLal, 1988). Earthworms play a vital role in soil fertility through the influence on water regime, aggregation, litter involment, soil organic dynamics and microbial activity (Briones et al., 1998).
Assessment of fipronil toxicity was designed to carry out with TLm (Median Tolerance Limit) test for 96 hrs.

A. Experimental Animal
Healthy and active Earthworm Eudrilus eugeniae having size of 6.51.5 cm were procured from the Ganpat University, Kherva village of Mahesana district, India. Before investigation Earthworm maintained at least for 10 days in laboratory floor containing dough of fertilizer. All dough of fertilizer were kept in the laboratory in cool place and covered with bag of jute and also use iron net to prevent the escape of animals and the foreign particles falling in. Dead specimens were removed immediately. Dough of fertilizer renewed every week. Chose different concentration like 17ml, 18ml, 19ml, and 20ml this concentration of solutions were prepared from the stock solution, and these are used for the bioassay test. The soil was mixed carefully with a glass rod.

B. Pesticides
The technical formulation of Fipronil (Lethal, SC 5%) a type of organophosphorus pesticide was selected for the study. The pesticide was purchased from the local pesticide supplier, Visnagar, Gujarat, India and used to check its impact on Earthworm.

C. Bioassay Test
The median tolerance limit (TLm-96 hrs) of Fipronil for the was Eudrilus eugeniae determined in the laboratory through static renewal bioassay test using standard method as described in APHA, (1995).
For the purpose of bioassay test, earthworms from the vermicompost (100gm) were used. For determining the LC50 selected 20 worms having similar size were inserted into each earthen port containing pesticide in different concentrations (ppm). First, the animals were exposed to different concentrations for range finding test where the lowest concentration at which 0% and the highest concentration for 50% mortality occurred.
No feed was given before 24 hrs and during the bioassay test. Test medium was renewed for every 24 hrs with their respective test concentration of the toxicant without aeration. Dead specimens were removed immediately as and when found mortality rate was recorded at the end of 24, 48, 72 and 96hrs. The median tolerance limit (TLm 96 hrs) was obtained by plotting graph with concentration series against the mortality rate for 96hrs. All the exposures were repeated at least three times to maintain the consistency in the results.

A. Bio-assay Test
Depicts the mortality and survival of Eudrilus eugeniae exposed to different concentration of Fipronil. Concentration at which minimum mortality (50%) Eudrilus eugeniae recorded was 17 ml and maximum concentration where 70% mortality noted was 20 ml. The exposure of Eudrilus eugeniae Fipronil treatment was most critical at 24 hrs.

B. Behavioral Responses
After application pesticides in soil instantly, animals were trying to jump out side of the earthen port. When the earthworm was exposed to various concentration of fipronil, they move immediately to the surface of the earthen port. The movement of earthworm was observed to be disrupted from 24 hrs of exposure. Irregular movement followed by imbalanced slide activity was observed. The sliding behavior was sudden, rapid non directed spurt of surface movement. The earthworm progressively showed signs of higher concentration (17, 18, 19, 20 ml) showed that animal lost their equilibrium and response to external stimuli such as touch by drowning to the soil trap and correspondingly died. Eudrilus eugeniae body color changes dark red to light red.  Table 1 Mortality of Eudrilus eugeniae exposed to Fipronil S. No.

Concentration of Fipronil in (ml)
No. of Earthworm Exposed Evaluated as toxicity grade of urea was categorized as "very toxic" to E. fetida. With consistent area of contact exposure of urea to the earthworm in different concentrations showed various toxic effects in filter paper substrate 28 μg/cm 2 by an average calculation. Thus, the comparative medium on the Petri plate bed. A geometric concentration series of test solution (5,10,20,40, and 80 mg/5 ml) was prepared and tested, in which mortality of earthworms was observed in 20, 40, and 80 mg concentrations after 18, 12, and 6 hr, respectively. Only the 5 mg/5 ml concentration was non-toxic to earthworm for 48 hr. The lethal concentration for 48 hr to E. fetida was 10 mg/5 ml concentration. The earthworm was not able to move around after 40 hr, neural retention and defoliation starts, and the body parts separate leading to the death. The lethal effect of the regularly used chemical fertilizer urea is killing the friend of farmer in a drastic way. In this preliminary screening experiment, the chemicals are shocked or absorbed into the earthworm body mainly through the outer dermal layer of skin when it is moving around the filter paper (Smetak et al., 2007, Estevez et al., 1996, Potter et al., 1985. The study found the organophosphorus pesticide chlorpyrifos is toxic to be earthworm species Eudrilus eugeniae at a concentration of 100 ppm, which showed 100 per cent mortality within 24 hours. Earthworm is one of the macro fauna indicator species. From the prohibit analysis high level of LC value can be seen from prohibit analysis so the study gives the information that the chlorpyrifos is toxic to earthworms at high concentrations levels from 100 ppm. L. rohita was silvery white in body in the control group throughout the experiment. The body colour changed from original silvery white to dark colour in pesticide treated fish. The fish maintained in freshwater behaved normal as body colour changed from original silvery white usual. But when the fish was exposed to pesticides monocrotophos and lambda cyhalothrin, erratic swimming, abnormal posture, disbalance, sluggishness, imbalance in posture, increase in surface activity, opercula movement, and gradual loss of equilibrium and spreading of excess of mucus all over the surface of the body were observed. A survey of LC50 values of different pesticides to the fish for different periods of exposure reveals the occurrence of a wide differences between duration of exposure and types of fishes (Macek and McAllister, 1970;Holden, 1972;Carter and Graves, 1973;Bakthavathasalam, 1980;Koundinya and Ramamurthi, 1980;Padmini, 1980;Rani et al., 1990;Dhanalakshmi, 1991;Sadhu, 1993;Pickering and Henderson, 1966;Santhakumar and Balaji, 2000;Mathivanan, 2004 andRamasamy et al., 2007). Changes in body colour have been reported in Anabas testudineusafter exposure to monocrotophos (Santhakumar and Balaji, 2000), C. punctatusto organophosphorus (Sandhu, 1993) and Cyprinuscarpioto ammonia stress (Israeli-weinstein and Kimmel, 1998).

Mortality in Numbers
The behavioral changes are considered directly related to complex physiological responses and have often been used as a sensitive indicator of stress (Little and Finger, 1990). The International Journal of Research in Engineering, Science and Management Volume-3, Issue-12, December-2020 https://www.ijresm.com | ISSN (Online): 2581-5792 109 change of body colour, behavioral changes such as irregular swimming movements, loss of equilibrium, restlessness and excess secretion of mucous suggest that L. rohita has undergone chemical stress when exposed to pesticide and the present study could be taken as an indicator of aquatic pollution. Result of LC50 96hr for both toxins showed that it decreases with increasing toxin concentration and duration of exposure. It means that by increasing the duration, lower concentrations of the toxins were required to kill 50% of fish and LC50values in the first 24 hours of the experiment were more than the end of the 96-hour LC50. Exposure time is one of the effective factors in OPs toxicity (Larkin & Tjeerdema, 2000). When fish are exposed to a constant concentration of toxins, their tolerance diminishes over time and the toxin has more time to affect. In toxic environments, fish exhibited irregular, erratic and darting swimming movements and loss of equilibrium which is due to inhibition of acetyl cholinesterase (AChE) activity leading to accumulation of acetylcholine in cholinergic synapses causing hyper stimulation.

Conclusion
After this experiment it was found that when pesticides mixing in soil. Within 24hrs, the effects are most pronounced on the earthworm. After 24hrs, the earthworms become stable. The pesticides concentration in the soil decreases so it does not have an effect on the earthworm after 24hrs-96hrs.
Finding of the work carried out during the experiment Eudrilus eugeniae different concentration of Fipronil which is used in agriculture to control the pests and indicated that the pesticide was lower toxic. Due to high humidity and low temperature, the effect of pesticides on the earthworm is less effect and low humidity and high temperature, the effect of pesticides on earthworm is more effect. Although the pesticide is more effective, the more effect on the earthworm. From the study, 50% mortality was observed in 17ml. The LC50 concentration were calculated from that it was clear that Fipronil is toxic to earthworm.