American-Eurasian J. Agric. & Environ. Sci., 5 (1): 31-38, 2009 ISSN 1818-6769 © IDOSI Publications, 2009 Corresponding Author: Hassan E. Abd-Elsalam, Mubarak City for Scientific Research and Technology Applications, New Borg El-Arab, Alexandria, Egypt E-mail: zamhelsayed@yahoo.com 31 Isolation and Identification of Three-Rings Polyaromatic Hydrocarbons (Anthracene and Phenanthrene) Degrading Bacteria Hassan E. Abd-Elsalam, Elsayed E. Hafez, Azhar A. Hussain, 1 2 3 Amany G. Ali and Amr A. El-Hanafy 4 5 Department of Environmental Biotechnology, 1 Genetic Engineering and Biotechnology Research Institute (GEBRI), Department of Plant Molecular Pathology, Arid Lands Cultivation Research Institute (ALCRI) 2 Microbiology and Botony Department, Women's College, Ain Shams University, Cairo, Egypt 3 Academy of Scientific Research and Technology Scholarship, Cairo, Egypt 4 Department of Nucleic Acid Research, GEBRI, 5 Mubark City for Scientific Researches and Technology Application, New Borg EL-Arab, Alexandria, Egypt Abstract: There are many industrial areas which produce large amount of poly-aromatic hydrocarbons (PAHs) in Egypt which represent severe hazards effects on the ambient environment. Bacterial strains were isolated from different contaminated sites in middle delta, Egypt and screened for PAHs degradation. Enrich media was used to isolate the anthracene and phenanthrene degrading bacteria. Fourteen bacterial isolates showed high degradation for both anthracene and phenantherene. For genotyping, these isolates were subjected to RAPD-PCR using four different primers. The data showed that the fourteen isolates were not related to each other. Only four isolates showed the highest ability for degradation were subjected to 16S rDNA sequence for identification. Partial sequence of 16S rDNA revealed that these isolates were Escherichia coli (EF105548), Soil bacterium (EF105549). Alcaligenes sp. (EF105546) and Thiobacter subterraneus (EF105547).The average degradation rates of anthracene by Escherichia coli (EF105548), Soil bacterium (EF105549), Alcaligenes sp. (EF105546) and Thiobacter subterraneus (EF105547) were 28.57, 30.19, 26.5875 and 32.11%, while those of phenanthrene were 42.45, 48.44, 34.35 and 40.45% for these strains, respectively. Key words:PAHs Anthracene Phenanthrene RAPD-PCR 16S rDNA E. coli Soil bacterium Alcaligenes sp. Thiobacter subterraneus INTRODUCTION organic compounds with two or more fused aromatic rings Environmental pollution was increased by increasing relatively low solubility in water, but are highly lipophilic the industry development all over the world and [2, 3]. Most of the PAHs with low vapor pressure in the air especially in Egypt; increment of these pollution caused are adsorbed on particles. When dissolved in water or many hazards for all organisms, even for humans such as adsorbed on particulate matter, PAHs can undergo carcinogenicity and toxicity. Also there has been photodecomposition when exposed to ultraviolet light increasing pollution with hydrocarbon compounds, many from solar radiation. In the atmosphere, PAHs can react of these hydrocarbons considered to be a potential health with pollutants such as ozone, nitrogen oxides and sulfur hazard [1, 2]. Some of hydrocarbon compounds pollutants dioxide, yielding diones, nitro- and dinitro-PAHs and are polycyclic aromatic hydrocarbons (PAHs). Polycyclic sulfonic acids, respectively. PAHs may also be degraded aromatic hydrocarbons (PAHs) are a large group of by some microorganisms in the soil [2, 4]. in linear, angular, or cluster arrangements. They have a