Imidacloprid is a second generation neonicotinoid pesticide (chloronicotinyl insecticide) that is used to treat plants against threatening pests. Application of imidacloprid as plant, seed and soil treatments has become a common practice to control pests on rice, maize, potatoes and vegetables. Besides controlling the pests in plants and soil, imidacloprid have come into direct or indirect contact to the nontargeted soil microorganisms and may affect their population and activity of microbial communities in soil. The aims of this research were the isolation and identification of imidacloprid degrading bacteria from soil samples in laboratory condition, and to determine effects of imidacloprid on the diversity of soil bacteria using suitable molecular markers. Several extraction methods have been published for determination of imidacloprid residues in different matrixes, but there is no record of Soxhlet extraction for imidacloprid residue analysis and comparison of its efficiencywith other extraction methods. Therefore reliable extraction method with appropriate use of organic solvent were optimized.