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Studies on Biodegradation of Pesticide from Wastewater Through Its Indigenous Microbial Flora

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Studies on Biodegradation of Pesticide from Wastewater Through Its Indigenous Microbial Flora STUDIES ON BIODEGRADATION OF PESTICIDE FROM WASTEWATER THROUGH ITS INDIGENOUS MICROBIAL FLORA SADIA SIRAJUDDIN Chapter 1 Introduction 1.1. Introduction Urbanization and extensive industrialization leads to the accumulation of a wide variety of pollutants and new technologies implanted to eradicate contaminants from the 1 STUDIES ON BIODEGRADATION OF PESTICIDE FROM WASTEWATER THROUGH ITS INDIGENOUS MICROBIAL FLORA SADIA SIRAJUDDIN environment. In agricultural field uses of pesticides has become necessary and their indiscriminate routine has inflicted serious environmental implication related to human health (Gavrilescu, 2005; Hussain et al., 2009). Pesticides intentionally made to be injurious and directly introduced into the environment from that only 5 percent reach to the target species and remaining enters into the environment. Presently, there are multiple possible methods for the clean-up of pesticides among them biodegradation proved to be most promising remedy for the treatment and detoxification of pesticides, particularly by the use of biocatalysts like enzymes, or by using micro-organisms as whole. Ghadiri, 2001; Bavcon et al., 2003; Nawab et al., 2003; Kodaka, et al., 2003; Sassman et al., 2004; Finley et al., 2010 discussed about the 2 STUDIES ON BIODEGRADATION OF PESTICIDE FROM WASTEWATER THROUGH ITS INDIGENOUS MICROBIAL FLORA SADIA SIRAJUDDIN pesticides degradation by the process of bioremediation as an substitute treatment approach that is effective, economical, , negligibly hazardous and environment-friendly. Schroll et al., (2004) stated that the degradation ability of microorganisms is directly proportional to their continuing adaptation to these compounds from the environment. Singh and Walker, (2006) further reported that indigenous microorganisms have the potential to be used for the bioremediation provides a cheap and efficient solution. According to Andreu and Pico, (2004) discussed about the pesticides are studied more than any other environmental contaminant. There is also increasing interest in their metabolites as transformation products can be present at higher levels with lower toxicity to biota in the environment than the parent pesticide itself (Nawab, et al., 3 STUDIES ON BIODEGRADATION OF PESTICIDE FROM WASTEWATER THROUGH ITS INDIGENOUS MICROBIAL FLORA SADIA SIRAJUDDIN 2003). However, metabolites also have the potential to produce an adverse impact on the environment (Sinclair and Boxall, 2003; Papadakis, and Papadopoulou-Mourkidou, 2002; Pozo et al., 2001). As a result, there is a need to identify metabolites during the process of pesticide detoxification. Potter and Wadkins, (2006); Redinbo and Potter (2005) stated that microbial enzymes advantageous for the detoxification of many pesticides. Specifically, carboxylesterases degrade many pesticides including pyrethroids (Wheelock et al., 2004; Stok et al., 2004a; Abernathy and Casida, 1973) and organophosphates (Casida and Quistad 2004; Kao et al., 1985) 4 STUDIES ON BIODEGRADATION OF PESTICIDE FROM WASTEWATER THROUGH ITS INDIGENOUS MICROBIAL FLORA SADIA SIRAJUDDIN Bass and Field, (2011) discussed about the enzyme families responsible for the degradation of pesticides that are Phosphotriesterases (Sogorb et al., 2004), Glutathione S-transferases (Enayati et al., 2005; Fournier et al., 1992) and Cytochrome P450 (Kulkarni and Hodgson 1984). 1.2. Objectives of the Study The present work is focused towards the use of efficient indigenous microbial strains and their extracellular enzyme for the degradation of the Malathion and Cypermethrin as a pilot study that can be used for commercial explications. Following are the main objectives which have been focused: 5 STUDIES ON BIODEGRADATION OF PESTICIDE FROM WASTEWATER THROUGH ITS INDIGENOUS MICROBIAL FLORA SADIA SIRAJUDDIN 1. To assess the physicochemical characteristics of wastewater of Lyari River that lead to the contamination of coastal ecosystem. The qualitative and quantitative analysis of wastewater samples have focused on parameters including pH, Temperature, DO -3 (Dissolve Oxygen), NH3 (Ammonia), TKN (Total Kjeldahl Nitrogen), PO4 (Phosphate), B0D5 (Biological Oxygen Demand), COD (Chemical Oxygen Demand), Total Coliform Count (TCC), Total Fecal Coliform Count(TFCC) and pesticide residues (malathion and cypermethrin) in order to compare range of concentration in accordance to the permissible limit provided by National Environmental Quality Standards (NEQS) for municipal and industrial effluent in Pakistan. (see in Chapter 2) 6 STUDIES ON BIODEGRADATION OF PESTICIDE FROM WASTEWATER THROUGH ITS INDIGENOUS MICROBIAL FLORA SADIA SIRAJUDDIN 2. Isolation, characterization and Polymerize chain reaction (PCR) based identification of indigenous bacteria capable for degrading malathion and cypermethrin. PCR was done for the amplification of the fragment of 16S rDNA gene of the DNA of isolated bacterial strain. The sequences were compared with the nucleotide database in GenBank and then aligned to construct a phylogenetic tree. This work as to describe the potential of pesticide degrading bacteria and to demonstrate eco- friendly technology suitable for wastewater containing pesticide. (see in Chapter 3) 3. Evaluation of Growth kinetics of metabolically versatile bacterial species namely Escherichia coli, Bacillus licheniformis, Pseudomonas aeruginosa by using 7 STUDIES ON BIODEGRADATION OF PESTICIDE FROM WASTEWATER THROUGH ITS INDIGENOUS MICROBIAL FLORA SADIA SIRAJUDDIN varying concentration of malathion and cypermethrin in minimal salt media. (see in Chapter 4) 4. Determination of the biodegradation kinetics of isolated metabolically versatile indigenous bacterial species namely Escherichia coli, Bacillus licheniformis, Pseudomonas aeruginosa under varying concentration of malathion and cypermethrin in minimal salt media. Gas chromatography was performed by using electron capture detector for the confirmation of biodegradation. (see in Chapter 5) 8 STUDIES ON BIODEGRADATION OF PESTICIDE FROM WASTEWATER THROUGH ITS INDIGENOUS MICROBIAL FLORA SADIA SIRAJUDDIN 5. To study degradation of malathion and cypermethrin by carboxylesterase of Escherichia coli IES-02 (KU593482) and analysis of novel product appearance over time by (GC- MS) gas chromatography mass spectrometry analysis. (see in Chapter 6) 1.3. Justification and Likely Benefits Around the world with the growth of agricultural activities malathion and cypermethrin are extensively used and discharged without any treatment into the environment caused seriously environmental implications because of their high toxicity. The problem is of special concern because of the development of resistance among bacterial strains to pesticides and the presence of pesticide metabolites into environment. Pesticide 9 STUDIES ON BIODEGRADATION OF PESTICIDE FROM WASTEWATER THROUGH ITS INDIGENOUS MICROBIAL FLORA SADIA SIRAJUDDIN resistance is mainly attributed by excessive and indiscriminate use of pesticides (Jayaraj, 1989). Gavrilescu, (2005) reported that the environmental pollution caused by pesticides spread over larger area. Therefore, the fate of pesticides pollution is often undefined. In fact, the damages are practically irreparable. To eliminate the antagonistic effect, it is important to treat them efficiently. Therefore, biodegradation is the best possible solution by transform organic matter into nutrients. Enzymes of microbial species break up complex compounds into metabolites which used by the microbial cells for reproduction and growth. Mixtures of species offer mineralization of particular pollutants completely because the metabolite of one species often serve as substrate to another species. 10 STUDIES ON BIODEGRADATION OF PESTICIDE FROM WASTEWATER THROUGH ITS INDIGENOUS MICROBIAL FLORA SADIA SIRAJUDDIN Enzymatic degradation of pesticides has two main advantages. Firstly, the degradation continues under relatively mild reaction conditions and secondly, the rate of reaction is faster than that by micro-organisms. Bacterial enzymatic detoxification of organophosphorus and pyrethroids pesticide has studied by many researchers, because it is economical, environment friendly and effective. Desaint et al., (2000); Pieper and Reineke, (2000) suggested that biodegradation are desirable to conventional methods because, microorganisms detoxify complex pollutants into less toxic metabolites. Carboxylesterases are major enzymes responsible for the catalysis of malathion and cypermethrin by the hydrolysis of carboxyl esters and also an important enzyme that deserve further study for applications in environmental monitoring. 11 STUDIES ON BIODEGRADATION OF PESTICIDE FROM WASTEWATER THROUGH ITS INDIGENOUS MICROBIAL FLORA SADIA SIRAJUDDIN Estimation of metabolites is also advantageous because transformation products can be present at higher levels with lower toxicity to biota in the environment than the parent pesticide itself. As a result, there is a need to identify metabolites during the process of pesticide detoxification. Furthermore kinetics of metabolites correctly describe the degradation of the parent compound. When the pesticide pollution is alarming remediation is obligatory to avoid relocation of pesticides to a more vulnerable areas. The identification of degradation products should facilitate an evaluation of the relative significance of chemical and microbial degradation in the environment. 12 STUDIES ON BIODEGRADATION OF PESTICIDE FROM WASTEWATER THROUGH ITS INDIGENOUS MICROBIAL FLORA SADIA SIRAJUDDIN 1.4. References Abernathy, C. O., & Casida,
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