Legionella pneumophila

Transcription

1 Legionella pneumophila / Legionella pneumophila - IMViC DNase lactamase Isolation & Identification of Legionella pneumophila from cooling towers May A.H. AL-Allaf Environmental & Pollution of Science Control Research Center Mosul university Amera M.M. Al-Rawi Department of Biology/ college Mosul university ABSTRACT The study included isolation of Legionella pneumophila from cooling water during March 2004 Septemper The bacterium was identified depending on ability to grow on selective media and microscopical examination which revealed that it is gram negative as well as biochemical tests were done and showed its variation reaction for

2 oxidase and positive reactions for catalase, motility, gelatinase, hippurate hydrolysis and starch hydrolysis as well as it was positive for -lactamase, DNase, Lipase and protease and it was negative for urease, carbohydrate fermentation, nitrate reductase and IMViC tests. Legionella Mathews & Roy, 2000.? Legionellaceae 20-2 (Addiss et al., 1989) Baker & Breach, 1980; Rodgers&Pasculle, 1991 ; Brooks et al., 2001) L-Cysteine HCl Bufferd Charcoal Yeast 7 3 Extract Agar (BCYE) (Tierney et al., 2003) L. pneumophila Cooling towers

3 (Bopp et al., BCYE Heat Treatment 1981) (0.2 ) (0.2 M) [(ph 2.2) (Forbes et al., 2002) Feeley Gorman Agar (FG) L-shape (Joklik Candle jar Growth factors CO 2? L- Cysteine HCl et al., 1980) Ferric pyrophosphate Ferric nitrate (Feeley et al., 1978,1979; Ristroph et al., 1980) (Behrman et al.,2004)

4 Modified Wadowsky Yee (MWY) Charcoal Yeast Extract Agar with Yeast Extract Broth Dyes Blood Agar Charcoal Yeast Extract Agar without Cysteine ( Hadgson & Casey, 1998 ; Brooks et al., 2001) (Hebert, 1981) (Prescott et al., 1996) (Thornsberry & Kirven, 1978) - Lactamase ( Collee et al., 1996 ; Koneman et al., 1997) IMViC Nutrient Gelatin Sulfide-Indol Motility SIM Starch Hydrolysis Hippurate Hydrolysis

5 DNase Agar Egg Yolk Agar Casein Hydrolysis Phenol Red Peptone Water Nitrate Broth Christensens Urea Agar Peptone Water Glucose Phosphate Peptone Water Simmon's Citrate Agar (MacFaddin, 1985 ; Collee et al., 1996 ) L. pneumophila Koid Casey Hadgson Heng

6 Industrial Water Society (IES) Wahala 200 Bopp Edelstien?? BCYE (Reinthaler et al., 1993; Ta et al., 1995) FG (Collee et al., 1996 ; Forbes et al., 2002) Feeley Charcoal Yeast Extract Hemoglobin Taylor Harrison

7 Weaver Agar Medium Edelstein MWY Vickers Ristroph Collee Tetramethyl-P-Phenyl Diamin Dihydro Chloride Brooks Forbes

8 Hebert - Lactamase Weaver Kirven Thornsberry - Lactamase Brooks DNase 2001 DNase Nitrate Reductase

9 Brenner IMViC L. pneumophila Tests ± Oxidase test Catalase test + Motility test + Gelatinase test + Hippurate hydrolysis test + Starch hydrolysis test + Lactamase test + Lipase test + DNase test + Protease test - Urease test - Carbohydrate Fermentation - Nitrate reductase - IMViC tests IMViC Variable ± Positive Negative

10 BCYE FG MWY

11 9 10 Control Control x -Lactamase

12 Addiss, D.G. ; Davis, J.P. and Laventure, M Community acquired Legionnaires disease associated with acooling tower. Evidence for longer-distance transport of Legionella pneumophila. Am. J. Epidemiol. 130: Baker, F.J. and Breach, M.R Medical Microbiological Techniques 1 st ed., Butterworths, London. p Behrman, R.E. ; Kliegman, R.M. and Jenson, H.B Nelson, Textbook of Pediatrics. 17 th ed. Saunders. An Imprint of Elsevier Science. p Bopp, C.A. ; Sumner, J.W. ; Morris, G.K., and Wells, J. G Isolation of Legionella spp. from environmental water samples by low-ph treatment and use of a selective. J. Clin. Microbiol. 13: Brenner, D.J Ten new species of Legionella. Intern. J. Syst. Bacteriol. 35: Brooks, G.F. ; Butel, J.S. and Morse, S.A Jawetz, Melinck, and Adclberg s, Medical Microbiology. 22 nd ed., Appetton and Lange, New York. p Collee, J.G. ; Fraser, A.G. ; Marmion, B.P. and Simmons, A Mackie and McCarteny Practical Medical Microbiology. 14 th ed., Churchill Livingston Inc. New York. p

13 Edelstein, P.H Comparative study of selective media isolation of Legionella pneumophila from potable water. J. Clin. Microbiol. 16: Feely, J.C. ; Gipson, R. J. and Gorman, G.W Charcoal yeast extract agar: Primary isolation media for Legionella pneumophila. J.Clin. Microbiol. 10: Feeley, J.C. ; Gorman, G.W. ; Weaver, R. E. ; Mackel, D.C. and Smith, H.W Primary isolation media for Legionnaires disease bacterium. J. Clin. Microbiol. 8: Forbes, A.B. ; Sahm, D.F. and Weissfeld, A. S Bailey and Scott s Diagnostic Microbiology. 9 th ed., Mosby. p Hadgson, J.M. and Casey, B.J Prevalence of Legionella bacteria in building water systems, article. Clayton Group Services. New York. 10: Hebert, G.A Hippurate hydrolysis by Legionella pneumophila. J. Clin. Microbiol. 13: Heng, B.H. ; Goh, K.T. ; Ng, D.L. and Ling, A.E Surveillance of Legionellosis and Legionella bacteria in the built environment in Singapore. Ann. Acad. Med. Singapore 26: Joklik, W.K. ; Willett, H.P. and Amos, D. B Zinsser Microbiology. 17 th ed. Appleton Century Crofts. New York. p Koid, M. ; Saito, A. ; Kusano, N. and Higa, F Detection of Legionella spp. in cooling tower water by the

14 polymerase chain reaction method. Appl. Environ. Microbiol. 59: Koneman, E.W. ; Allen, S.D. ; Janda, W.M. ; Screchenberger, D.C. and Winn, W. C Color Atlas and Textbook of Diagnostic Microbiology 5 th ed., Lippincott- Raven Publishers, Philadelphia, USA. p MacFaddin, J.F Biochemical Tests for Identification of Medical Bacteria. 2 nd Ed., Waverly Press. Inc. Baltimore. USA. Mathews, M. and Roy, C.R Identification and subcellular localization of the Legionella pneumophila. Icm protein : a factor essential for establishment of a replicative organelle in eukaryoticl host cells. Infect. Immun. 68: Prescott, L. M. ; Harley, J. P. and Klein, D.A Microbiology. 3 rd ed., Wm C. Brown Communication, Inc. Iowa, USA. Reinthaler, F.F. ; Sattler, J. ; Schaffler-Dulling, K. ; Weinmayr, B. and Marth, E Comparative study of procedures for isolation and cultivation of Legionella pneumophila from tap water in hospitals. J. Clin. Microbiol. 31: Ristroph, J.D. ; Hedlund, K.W. and Allen, R. G Liquid for growth of Legionella pneumophila. J. Clin. Microbiol. 11: Rodgers, F.G. and Pasculle, A.W Legionella. In Balows A, Hausler, W.J. Jr., Herrmann, K.L. (eds.). Manual of Clinical Microbiology, Washington DC, Am. Soc. Microbiol. p

15 Ta, A.C. ; Stout, J.E. ; Yu, V.L. and Wagener, M.M Comparison of culture methods for monitoring Legionella species in hospital potable water systems and recommendations for standardization of such methods. J. Clin. Microbiol. 33: Thornsberry, C. and Kirven, L.A Lactamase of the Legionnaires bacterium. Curr. Microbiol. 1: Tierney, L.M. ; McPhee, S.J. and Papadakis, M.A Current Medical Diagnosis and Treatment 42 nd ed., Lange Medical Books. McGraw-Hill. New York. p Vickers, R.M. ; Brown, A. and Garrity, G.M Dye-containing buffered charcoal yeast extract for the differentiation of members of the family Legionellaceae. J. Clin. Microbiol. 13: Wahala, W.M First isolation of Legionella pneumophila in Srilanka. Ceylon J. Med. 45 : Weaver, R.E Cultural and staining characteristics. In : Jones, G.L. ; Hebert G.A. (eds.). Legionnaires ; the Disease Bacteriology and Methodology. Centers for Disease, Control, Atlanta, Georgia,p

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