Comparison of BCYE and BMPA media on recovery rate of Legionella pneumophila

  • Lucky Moehario
  • Enty Tjoa
  • Mutiara J. Taslim
  • Yohanna Angelina
Keywords: Legionella pneumophila, specific medium, BCYE, BMPA, recovery rate


Latar belakang: Legionella pneumophila (L. pneumophila) telah banyak diketahui sebagai penyebab legionellosis; habitat nya di berbagai sumber air, lingkungan lembab dan hangat. Metode kultur menggunakan media spesifik masih menjadi baku emas untuk identifikasi L. pneumophila. Penelitian ini bertujuan membandingkan recovery rate L. pneumophila ATCC®33823 pada medium spesifik BCYE, dan medium selektif BMPA.

Metode: Dilakukan dilusi serial pada suspensi 0,5 McFarland L. pneumophila ATCC®33823; 100 ul dari tiap tingkat dilusi diinokulasi pada medium Buffered Charcoal Yeast Extract (BCYE) dan medium BMPA (medium BCYE yang ditambahkan suplemen BMPA-α), secara duplo. Konsentrasi suspensi dihitung menggunakan metode Angka Lempeng Total (ALT) dari Standar Nasional Indonesia No. 01-2332.3-2006. Persentase recovery rate dihitung dan dianalisis secara statistik menggunakan SPSS versi 23,0.

Hasil: Jumlah koloni L. pneumophila yang tumbuh pada medium BMPA jauh lebih tinggi daripada yang tumbuh di medium BCYE; konsentrasi tertinggi yang diperoleh adalah pada medium BMPA sebesar 1,45 x 107 CFU/ml. Persentase recovery rate pada medium BMPA adalah 96,67%, dan 60,67% pada medium BCYE.

Kesimpulan: Recovery rate medium BMPA untuk pertumbuhan koloni L. pneumophila ATCC®33823 jauh lebih tinggi daripada media BCYE, karena itu medium BMPA dapat direkomendasikan untuk kultivasi L. pneumophila, khususnya pada program surveilans berbagai sumber air dengan biaya lebih terjangkau.

Kata kunci: Legionella pneumophila, medium spesifik, BCYE , BMPA, recovery rate



Background: Legionella pneumophila (L. pneumophila) has been known as the etiology of legionellosis; they live in aquatic environment, warm and moist. Culture method using specific medium remains as the gold standard in the identification of L. pneumophila. This study aimed to compare the recovery rate of L. pneumophila ATCC® 33823 on the specific medium BCYE for the cultivation of Legionella, and BMPA, the selective medium.

Methods: Suspension of L. pneumophila ATCC® 33823 of 0.5 McFarland was diluted to 10 fold serial dilution; 100 ul of each dilution was inoculated on Buffered Charcoal Yeast Extract (BCYE) medium, and BMPA (BCYE supplemented with BMPA-α) in duplicate manner. The concentration was calculated using Total Plate Count standard as of Indonesian Nasional Standard number 01-2332.3-2006. The percentage of recovery rate was calculated, and the statistical analysis was performed using SPSS version 23.0.


Results: Numbers of colonies of L. pneumophila grew on BMPA was much higher than on BCYE medium; the highest concentration was yielded on BMPA medium i.e. 1.45x107 CFU/ml. The recovery rates were 96.67% and 60.67% on BMPA medium and BCYE subsequently.

Conclusion: The recovery rate of the BMPA medium on the colony growth of L. pneumophila ATCC®33823 was markedly higher than the BCYE, therefore BMPA medium can be suggested to be used in the cultivation of L. pneumophila especially in the routine surveillance program for water sources with less cost.

Keywords: Legionella pneumophila, specific medium, BCYE , BMPA, recovery rate


CDC [Internet]. About legionnaires disease and pontiac fever. 2019. [Cited 2020 May 25]. Available from:

Farver CF. Bacterial Diseases. Pulmonary Pathology [Internet]. 2nd ed. Philadelphia, PA: Elsevier; 2018. [Cited 2018 Sep 16] p 163–200. Available from:!/content/book/3-s2.0 B978032 3393089000108?scrollTo=%23hl0001580.

Yackley JK. Notes from the Field: Legionellosis outbreak associated with a hotel aquatics facility - Tennessee, 2017. [Cited 2019 Dec 6]. MMWR Morb Mortal Wkly Rep [Internet]. 2018; 67. Available from: .

Edelstein PH. Legionnaires’ disease, pontiac fever, and related illnesses. In: Feigin and Cherry’s Textbook of Pediatric Infectious Diseases [Internet]. 8th ed. Philadelphia, PA: Elsevier; 2019. [Cited 2018 Sep 18] p 1228-1235. e4. Available from:!/content/book/3-s2.0-B9780323376921001374? scrollTo=%23hl0000638.

Rubin LG. Legionella Species. Principles and Practice of Pediatric Infectious Diseases [Internet]. 5th ed. Philadelphia, PA: Elsevier; 2018 [Cited 2018 Sep 16]; p. 948-952.e1. Available from: https://www.clinicalkey. com/#!/content/book/3-s2.0-B9780323401814001778?scrollTo=%23hl0000336.

Byrne BG, McColm S, McElmurry SP, Kilgore PE, Sobeck J, Sadler R, et al. Prevalence of infectioncompetent serogroup 6 Legionella pneumophila within premise plumbing in Southeast Michigan. mBio. 2018 Mar 7; 9(1): e00016-18.

Al-Matawah Q, Al-Zenki S, Al-Azmi A, Al-Waalan T, Al-Salameen F, Hejji AB. Legionella detection and subgrouping in water air-conditioning cooling tower systems in Kuwait. Environ Sci Pollut Res Int. 2015 Jul; 22(13): 10235–41.

Aksono EB, Farahdiba AA, Hestianah EP. Legionella pneumophila bacteria detected in swimming pool water of Surabaya by using nested Polymerase Chain Reaction. J Vet. 2017; 18(2): 221–5.

Yasmon A, Yusmaniar Y, Anis A, Bela B. Simultaneous detection of Legionella species and Legionella

pneumophila by duplex PCR (dPCR) assay in cooling tower water samples from Jakarta, Indonesia. Med J Indones. 2010 Nov 1; 19(4): 223–7.

Ministry of Health Republic of Indonesia. Minister of Health Decree No. 1538/Menkes/SK/1/2003 about Standards of Legionella Specimen Management. Jakarta : Ministry of Health of Indonesia. 2003.

Sikora A, Wojtowicz-Bobin M, Koziol-Montewka M, Magrys A, Gladysz I. Prevalence of Legionella pneumophila in water distribution systems in hospitals and public buildings of the Lublin region of eastern Poland [Internet] [Cited 2020 May 25]. Available from: distributionsystems-in-hospitals and,72258,0,2.html/

D’Alessandro D, Fabiani M, Cerquetani F, Orsi G. Trend of legionella colonization in hospital water supply. Ann Ig Med Prev E Comunità. 2015 Jun 5; 27: 460–6.

Prussin AJ, Schwake DO, Marr LC. Ten questions concerning the aerosolization and transmission of Legionella in the built environment [Internet]. [Cited 2020 May 25]. Available from: https://www.ncbi.nlm.

Ministry of Health Republic of Indonesia. Minister of Health Decree No_7_Th_2019_ttg_Kesehatan_ Lingkungan_Rumah_Sakit.pdf [Internet]. [Cited 2019 Oct 29]. Available from: http://aspak.yankes.kemkes. Rumah_Sakit.pdf.

Agarwal S, Abell V, File TM. Nosocomial (Health Care–Associated) legionnaire’s disease. Infect Dis Clin North Am. 2017 Mar; 31(1): 155–65.

Mercante JW, Winchell JM. Current and emerging Legionella diagnostics for laboratory and outbreak investigations. Clin Microbiol Rev. 2015 Jan; 28(1):95–133.

Kotrbancová M, Špaleková M, Fulová M, Trnková K, Perželová J. Legionellosis and its diagnosis. Epidemiol Mikrobiol Imunol Cas Spolecnosti Epidemiol Mikrobiol Ceske Lek Spolecnosti JE Purkyne [Internet]. 2017;66(3): 133–9. [Cited 2020 May 25]. Available from:

Giglio O, Diella G, Trerotoli P, Consonni M, Palermo R, Tesauro M, et al. Legionella detection in water networks as per ISO 11731:2017: can different filter pore sizes and direct placement on culture media influence laboratory results? Int J Environ Res Public Health. 2020 Mar 20;17: 1-8.

CDC [Internet]. Monitoring your building water for Legionella. 2019. [Cited 2020 May 25]. Available from: html.

Whelen C. Legionella and Bordetella. Textbook of diagnostic microbiology. 6th ed. Philadelphia: Saunders; 2018. p. 401–7.

Caroll KC, Brooks GF, Jawetz E, Melnick JL, Adelberg EA (ed). Bacteriology. Jawetz, Melnick, & Adelberg’s medical microbiology. 28th ed. New York: McGraw Hill Medical; 2019. p. 315–8.

Pierre DM, Baron J, Yu VL, Stout JE. Diagnostic testing for Legionnaires’ disease. Ann of Clin Microbiol and Antimicrob [Internet]. 2017 August 29 [Cited 2020 May 24]; 16 (59) : 1-4. Available from: https://annclinmicrob.

Veenendaal HR, Brouwer-Hanzens AJ, van der Kooij D. Incubation of premise plumbing water samples on Buffered Charcoal Yeast Extract at elevated temperature and pH selects for Legionella pneumophila. Water Res. 2017 15; 123: 439–47.

Engl ards unit NISPH, st 61 Colindale Avenue London NW9 5EQ Email, st. UK Standards for Microbiology Investigation: ID 18 Identification of Legionella species - GOV.UK. Public Health Engl. (03): 09–13. Public Health England. (2015).

Badan Standardisasi Nasional. Cara uji mikrobiologibagian 3: penentuan angka lempeng total (ALT) pada produk perikanan. Standar Nasional Indonesia. Jakarta; 2006. Indonesian.

Roelofsen E, Leeuwen MV, Meijer-Severs GJ, Wilkinson MHF, Degener JE. Evaluation of the effects of storage in two different swab fabrics and under three different transport conditions on recovery of aerobic and anaerobic bacteria. Journal of Clinical Microbiology [Internet]. 1999 [Cited 2019 October 30]; 37(9): 3041-3. Available from:

Catherine AB, Edelstein PH. Precision and accuracy of recovery of Legionella pneumophila from seeded tap water by filtration and centrifugation. Applied and Environmental Microbioogyl [Internet]. 1995 [Cited 2019 November 21]; 61(5):1805–9. Available from:

Descours G, Cassier P, Forey F, Ginevra C, Etienne J, Lina G, et al. Evaluation of BMPA, MWY, GVPC and BCYE media for the isolation of Legionella species from respiratory samples. J Microbiol Methods [Internet]. 2014 Mar 1; 98: 119–21. Available from: 10.1016/j. mimet.2014.01.001.

Edelstein PH. Improved semiselective medium for isolation of Legionella pneumophila from contaminated clinical and environmental specimens. J Clin Microbiol [Internet]. 1981 Sep [Cited 2018 August 16];14(3):298–303. Available from:

Council of Europe. Microbial examination of non-sterile products: microbial enumeration tests. Pharmacopeia. 2018.

Fliermans CB, Cherry WB, Orrison H, Smith SJ, Tison DL, Pope DH. Ecological niche of Legionella pneumophila. Applied and Environmental Microbiology. 1981;41(1):9-16.

Moehario LH, Robertus T, Grace Y, Tjoa E. Screening of Legionella pneumophila from water sources in the hospitals in Jakarta. Health Science Journal of Indonesia [Internet]. 2019 [Cited 2018 August 15]; 10(1):21–6. Available from:

How to Cite
Moehario, L., Tjoa, E., Taslim, M., & Angelina, Y. (2020). Comparison of BCYE and BMPA media on recovery rate of Legionella pneumophila. Health Science Journal of Indonesia, 11(1), 32-37.

Most read articles by the same author(s)