Isolation and identification of Legionella spp. from different aquatic sources in south-west of Iran by molecular &culture methods

Document Type: Original article


Department of Pathobiology, School of Veterinary Medicine, Shiraz University, Shiraz, Iran


Legionella pneumophila, the causative agent of Legionnaires’ diseases (LD) is usually transmitted to humans via inhalation of aerosols from contaminated natural and manmade water sources. These organisms may become fatal especially in immunocompromised patients and LD is the one of the important disease from a public health perspective. This survey investigated the frequency of Legionella spp. including L. pneumophila, in some cold and warm water systems in South-West of Iran by culture and PCR methods. Thirty four water samples were collected from diverse water supply systems. After acid and heat treatments of samples, inoculated onto buffered charcoal yeast extract agar. Isolated colonies were confirmed by morphological and biochemical tests. Then the isolates were examined for icmO, sidA and lidA genes by PCR assay. This study showed that frequency of L. pneumophila was 4 by culture and 14 by PCR. PCR method to be efficient and sensitive test for rapid detection of these organisms in environmental water sources. This study emphasizes the need for effective infection control and prevention strategies to minimize the risk from exposure to potential pathogens such as Legionella spp. and to create a safe working environment.


1. Cho MS, Ahn TY, Joh K, Lee ES, Park DS. Improved PCR assay for the species-specific identification and quantitation of Legionella pneumophila in water. Appl Microbiol Biotechnol 2015;99:9227-9236.

2. Ehrhardt J, Alabi AS, Kuczius T, Tsombeng FF, Becker K, Kremsner PG, Schaumburg F, Esen M. Population structure of Legionella spp. from environmental samples in Gabon, 2013. Infect Genet Evol 2013;33:299-303.

3. Moosavian M, Dashti A. Isolation and identification of legionellosis agents from fish ponds, swimming pools and cooling towers in Khuzestan province, Iran. Jundishapur J Microbiol 2011;4:209-215.

4. Li L, Qin T, Li Y, Zhou H, Song H, Ren H, Li L, Li Y, Zhao D. Prevalence and molecular characteristics of waterborne pathogen Legionella in industrial cooling tower environments. Int J Environ Res Public Health 2015;12:12605-12617.

5. Li L, Mendis N, Trigui H, Faucher SP. Transcriptomic changes of Legionella pneumophila in water. BMC Genomics 2015;26:616-637.

6. Dimitriadi D, Velonakis E. Detection of Legionella spp. from domestic pater in the Prefecture of Arta, Greece. Journal of Pathogens2014;10:1-5.

7. Khemiri A, Lecheheb SA, Chi-Song PC, Jouenne T, Cosette P. Proteomic regulation during Legionella pneumophila biofilm development: decrease of virulence factors and enhancement of response to oxidative stress. J Water Health 2014;12:242-253.

8. Khalegi M, Navidpour L, Sepehrizadeh Z, Nazari P, Pourmand MR, Samadi N, Moosavian SM, Shahverdi AR. The sensitization of Legionella pneumophila to some antibiotics by reserpine and anti-Legionella effects of different benzofuranone derivatives. J Med Bacteriol2013;2:35-40.

9. Newton HJ, Ang DKY, van-Driel IR, Hartland EL. Molecular pathogenesis of infections caused by Legionella pneumophila. J Clin Microbiol 2010;2:274-298.

10. Flanders WD, Kirkland KH, Shelton BG. Effects of holding time and measurement error on culturing Legionella in environmental water samples. Water Res 2014;62: 293-301.

11. Chaudhry R, Valavane A, Mohan A, Dey AB. Legionella pneumophila infection associated with renal failure causing fatality in a known case of sarcoidosis. Indian J Med Microbiol 2014;32:324-327.

12. Heijnsbergen EV, Husman DR, Lodder WJ, Bouwknegt M, Van-Leeuwen AE, Bruin JP, Euser SM, Boe D, Schalk JAC. Viable Legionella pneumophila bacteria in natural soil and rain water puddles. J Apple Microbiol 2014;117:882-890.

13. Gilmour MW, Bernard K, Tracz DM, Olson AB, Corbett CR, Burdz T, Ng B, Wiebe D, Broukhanski G, Boleszczuk P, Tang P, Jamieson F, Domselaar GV, Plummer FD, Berry JD. Molecular typing of a Legionella pneumophila outbreak in Ontario, Canada.J. Med. Microbiol 2007;56:336–341.

14. Movahedian-Attar H, Shahmansouri MR, Neshat AA, Fazeli M. Identification of Legionella in the hot water supply of a general hospital in Isfahan. J Res Med Sci 2004;6:289-293.

15. D’Auria G, Jiménez-Hernandez N, Peris-Bondia F, Moya A, Latorre A. Legionella pneumophila pangenome reveals strain-specific virulence factors. BMC Genomics 2010;11:181-194.

16. Baghalasghari F, Nikaeen M, Hatamzadeh M, Hassanzadeh A. Surveillance of Legionella species in hospital water systems: the significance of detection method for environmental surveillance data. J. Water Health 2013;11:713-719.

17. Gholipour A, Moosavian M, Galehdari H. Cloning and periplasmic expression of peptidoglycan-associated lipoprotein (PAL) protein of Legionella pneumophila in Escherichia coli.Jundishapur J Microbiol 2010;3:1–9.

18. Collins S, Jorgensen F, Willis C, Walker J. Real-time PCR to supplement gold-standard culture-based detection of Legionella in environmental samples. J Appl Microbiol2015; 27:911-916.

19. Ahmadinejad M, Shakibaie MR, Shams K, Khalili M. Detection of Legionella pneumophila in cooling water systems of hospitals and nursing homes of Kerman city. Int J Biol Sci 2011; 52:799-802.

20. Ghotaslou R, YeganehSefidan F, Akhi MT, Soroush MH, Hejazi MS. Detection of Legionella contamination in Tabriz hospitals by PCR assay.Advanced Pharmaceutical Bulletin 2013; 3:131-134.

21. Mouchtouri VA, Rudge JW. Legionnaires’ disease in hotels and passenger ships: a systematic review of evidence, sources, and contributing factors. J Travel Med 2015; 1195-1982; 325-337.