{"references": ["R. Tello, G, Harthell, P. Costello, K. Slokes. \"Legionella infected\nthoracic aortic graft,\" Comput Med Imaging Graph. 17(1), 1993, pp. 61-\n77.", "P. Villari, E. Motti, C. Farullo, I, Torre. \"Comparison of conventional\nculture and PCR methods for the detection of Legionella pneumophila in\nwater,\" Lett Appl Microbiol. 27, 1998, pp. 106-110.", "F. F. Reinthaler, J. Sattler, K. Schaffler-Dullnig, B. Weinmayr, E. Marth,\n\"Comparative Study of Procedures for Isolation and Cultivation of\nLegionella pneumophila from Tap Water in Hospitals\" Journal of\nClinical Microbiology. 31(5), 1993, pp. 1213-1216.", "W. S. Shadrach, K. Rydzewski, U. Laube, G. Holland, M. \u251c\u00fbzel, A. F.\nKiderlen, A. Flieger, \"Balamuthia mandrillaris, Free-Living Ameba and\nOpportunistic Agents of Encephalitis, Is Potential Host for Legionella\npneumophila Bacteria\" Applied and Environmental Microbiology. 71(5),\n2005, pp..2244-2249.", "A. C. England, D.W. Fraster,. \"Sporadic and epidemic nasocomial\nlegionellosis in the United States\" Epidemiology features. Am J Med 70\n(3), 1981, pp. 707-711.", "R. C. Mahon, G. J. Manuselis, Diagnostic Microbiology. WB Saunders\nCo London, 1995, pp. 430-439.", "S. Buchbinder, K. Trebesius, J. Heesemann, \"Evaluation of detection of\nLegionella SPP. In water samples by fluorescence in situ hybridization,\nPCR amplification and bacterial culture\" Int J Med Microbiol 292, 2002.\npp. 241-145.", "H. Miyamoto, H. Yamamoto, K. Arima, J. Fujii, K. Maruta, K. Izu, T.\nShiomori, S. Yoshida, \"Development of a New Seminested PCR Method\nfor Detection of Legionella Species and Its Application to Surveillance\nof Legionellae in Hospital Cooling Tower Water\". Applied and\nEnvironmental Microbiology 63(7), 1997, 2489-2494.", "H. Yamamoto, Y. Hashimoto, T. Ezaki, \"Comparison of detection\nmethods for Legionella species in environmental water by colony\nisolation fluorescent antibody staining and polymerase chain reaction\"\nMicrobiol Immunol. 37, 1993 pp. 617-622.\n[10] A. H. Movahedian, M. R. Shahmansouri, A. A. Neshat, M. Fazeli,\n\"Identification of Legionella in the hot water supply of a General\nHospital in Isfahan\" J. of Research in Medical Sciences 6,\n2004, pp. 40-46.\n[11]N. Wellinghausen, C. Frost, R. Marre, \"Detection of\nLegionellae in Hospital Water Samples by Quantitative\nReal Time Light Cycler PCR,\" Applied and Environmental\nMicrobiology. 67(9), 2001, pp. 3985-3993.\n[12] G. F. Brooks, E. Jawetz, J. I. Melnick, E. A. Adelberg, \"Legionella\npneumophila In Medical Microbiology\" 19th Edition. Appleton. 1991,\npp. 263-266.\n[13] M. M. Aslani, M. Pourmansour, M. Motavalian, \"Isolation of Legionella\npneumophila from Tehran Hospital water samples,\" Iranian\nBiochemical Journa,l 1(1), 1997, pp. 65-67.\n[14] M. Moosavian, B. Fathollahzadeh, K. Amoli, \"Isolation of Legionella\npneumophila Serogroups 1, 8 & 10 (Causative Agents of Legionnairs\nDisease) From Water Sources in Tehran, Iran,\" Iranian Biomedical\nJournal. 2(2), 1998, pp. 83-87.\n[15] R. Hosseini-Doust S David, \"Isolation of Legionella from Hospitals\nWater System,\" Kosar Medical Journal. 3(3), 1998, 145-150.\n[16] F. F. Reinthaler, J. Sattler, K. Schaffler-Dullnig, Comparative Study of\nProcedures for Isolation and Cultivation of Legionella pneumophila\nfrom Tap Water in Hospitals. Journal of Clinical Microbiology, May\n1993, pp 1213 - 1216."]}

Legionella pneumophila is involved in more than 95% cases of severe atypical pneumonia. Infection is mainly by inhalation the indoor aerosols through the water-coolant systems. Because some Legionella strains may be viable but not culturable, therefore, Taq polymerase, DNA amplification and semi-nested-PCR were carried out to detect Legionella-specific 16S-rDNA sequence. For this purpose, 1.5 litter of water samples from 77 water-coolant system were collected from four different hospitals, two nursing homes and one student hostel in Kerman city of Iran, each in a brand new plastic bottle during summer season of 2006 (from April to August). The samples were filtered in the sterile condition through the Millipore Membrane Filter. DNA was extracted from membrane and used for PCR to detect Legionella spp. The PCR product was then subjected to semi-nested PCR for detection of L. pneumophila. Out of 77 water samples that were tested by PCR, 30 (39%) were positive for most species of Legionella. However, L. pneumophila was detected from 14 (18.2%) water samples by semi-nested PCR. From the above results it can be concluded that water coolant systems of different hospitals and nursing homes in Kerman city of Iran are highly contaminated with L. pneumophila spp. and pose serious concern. So, we recommend avoiding such type of coolant system in the hospitals and nursing homes.