The Burkholderia cepacia complex comprises groups of genomovars (genotypically distinct strains with very similar phenotypes) that have emerged as important opportunistic pathogens in cystic fibrosis (CF) patients. The inflammatory response against bacteria in the airways of CF individuals is dominated by polymorphonuclear cells and involves the generation of oxidative stress, which leads to further inflammation and tissue damage. Bacterial catalase, catalase-peroxidase and superoxide dismutase activities may contribute to the survival of B. cepacia following exposure to reactive oxygen metabolites generated by host cells in response to infection. In the present study the authors investigated the production of catalase, peroxidase and SOD by isolates belonging to various genomovars of the B. cepacia complex. Production of both catalase and SOD was maximal during late stationary phase in almost all isolates examined. Native PAGE identified 13 catalase electrophoretotypes and two SOD electrophoretotypes (corresponding to an Fe-SOD class) in strains belonging to the six genomovars of the B. cepacia complex. Seven out of 11 strains displaying high-level survival after H2O2 treatment in vitro had a bifunctional catalase/peroxidase, and included all the genomovar III strains examined. These isolates represent most of the epidemic isolates that are often associated with the cepacia syndrome. The majority of the isolates from all the genomovars were resistant to extracellular \(O_{2}^{{-}}\) , while resistance to intracellularly generated \(O_{2}^{{-}}\) was highly variable and could not be correlated with the detected levels of SOD activity. Altogether the results suggest that resistance to toxic oxygen metabolites from extracellular sources may be a factor involved in the persistence of B. cepacia in the airways of CF individuals.
Bals, R., Weiner, D. J. & Wilson, J. M.(1999). The innate immune system in cystic fibrosis lung disease. J Clin Invest103, 303-307.[CrossRef][Google Scholar]
Bandyopadhyay, P. & Steinman, H. M.(1998).Legionella pneumophila catalase-peroxidases: cloning of the katB gene and studies of KatB function. J Bacteriol180, 5369-5374.
[Google Scholar]
Bannister, J. V. & Calabrese, L.(1987). Assays for superoxide dismutase. Methods Biochem Anal32, 279-312.
[Google Scholar]
Barnes, A. C., Horne, M. T. & Ellis, A. E.(1996). Effect of iron on expression of superoxide dismutase by Aeromonas salmonicida and associated resistance to superoxide anion. FEMS Microbiol Lett142, 19-26.[CrossRef][Google Scholar]
Barnes, A. C., Balebona, M. C., Horne, M. T. & Ellis, A. E.(1999). Superoxide dismutase and catalase in Photobacterium damselae subsp. piscidia and their roles in resistance to reactive oxygen species. Microbiology145, 483-494.[CrossRef][Google Scholar]
Beauchamp, C. & Fridovich, I.(1971). Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. Anal Biochem44, 276-287.[CrossRef][Google Scholar]
Brown, S. M., Howell, M. L., Vasil, M. L., Anderson, A. J. & Hassett, D. J.(1995). Cloning and characterization of the katB gene of Pseudomonas aeruginosa encoding a hydrogen peroxide-inducible catalase: purification of KatB, cellular localization, and demonstration that it is essential for optimal resistance to hydrogen peroxide. J Bacteriol177, 6536-6544.
[Google Scholar]
Burkholder, W. H.(1950). Sour skin, a bacterial rot of onion bulbs. Phytopathology40, 115-117.
[Google Scholar]
Burns, J. L., Jonas, M., Chi, E. Y., Clark, D. K., Berger, A. & Griffith, A.(1996). Invasion of respiratory epithelial cells by Burkholderia (Pseudomonas) cepacia. Infect Immun64, 4054-4059.
[Google Scholar]
Carlioz, A. & Touati, D.(1986). Isolation of superoxide dismutase mutants in Escherichia coli: is superoxide dismutase necessary for aerobic life? EMBO J5, 623-630.
[Google Scholar]
Cline, M. J.(1975). Chemotaxis, phagocytosis and microbial killing. In The White Cell , pp. 71-82. Edited by M. J. Cline. Cambridge, MA:Harvard University Press.
De Groote, M. A., Ochsner, U. A., Shiloh, M. U. & 7 other authors (1997). Periplasmic superoxide dismutase protects Salmonella from products of phagocyte NADPH-oxidase and nitric oxide synthase. Proc Natl Acad Sci USA94, 13997–14001.[CrossRef][Google Scholar]
Fang, F. C., DeGroote, M. A., Foster, J. H. & 8 other authors (1999). Virulent Salmonella typhimurium has two periplasmic Cu,Zn-superoxide dismutases. Proc Natl Acad Sci USA96, 7502–7507.[CrossRef][Google Scholar]
Forehand, J. R., Johnston, R. B. J. & Bomalaski, J. S.(1993). Phospholipase A2 activity in human neutrophils. Stimulation by lipopolysaccharide and possible involvement in priming for an enhanced respiratory burst. J Immunol151, 4918-4925.
[Google Scholar]
Franzon, V. L., Arondel, I. & Sansonetti, P. I.(1990). Contribution of superoxide dismutase and catalase activities to Shigella flexneri pathogenesis. Infect Immun58, 529-535.
[Google Scholar]
Gillis, M., Van, T. V., Bardin, R. & 7 other authors (1995). Polyphasic taxonomy in the genus Burkholderia leading to an emended description of the genus and proposition of Burkholderia vietnamiensis sp. nov. for N2-fixing isolates from rice in Vietnam. Int J Syst Bacteriol45, 274–289.[CrossRef][Google Scholar]
Govan, J. R. W. & Deretic, V.(1996). Microbial pathogenesis in cystic fibrosis: mucoid Pseudomonas aeruginosa and Burkholderia cepacia. Microbiol Rev60, 539-574.
[Google Scholar]
Govan, J. R. W. & Vandamme, P.(1998). Agricultural and medical microbiology: a time for bridging gaps. Microbiology144, 2373-2375.[CrossRef][Google Scholar]
Govan, J. R., Hughes, J. E. & Vandamme, P.(1996).Burkholderia cepacia: medical, taxonomic and ecological issues. J Med Microbiol45, 395-407.[CrossRef][Google Scholar]
Heikkila, R. E. & Cabbat, F.(1976). A sensitive assay for superoxide dismutase based on the autoxidation of 6-hydroxydopamine. Anal Biochem75, 356-362.[CrossRef][Google Scholar]
Hughes, J. E., Stewart, J., Barclay, G. R. & Govan, J. R.(1997). Priming of neutrophil respiratory burst activity by lipopolysaccharide from Burkholderia cepacia. Infect Immun65, 4281-4287.
[Google Scholar]
Imlay, J. A. & Linn, S.(1988). DNA damage and oxygen radical toxicity. Science240, 1302-1309.[CrossRef][Google Scholar]
Katsuwon, J. & Anderson, A. J.(1989). Response of plant-colonizing pseudomonads to hydrogen peroxide. Appl Environ Microbiol55, 2985-2989.
[Google Scholar]
Katsuwon, J. & Anderson, A. J.(1992). Characterization of catalase activities in a root-colonizing isolate of Pseudomonas putida. Can J Microbiol38, 1026-1032.[CrossRef][Google Scholar]
Kelley, T. G. & Drumm, M. L.(1998). Inducible nitric oxide synthase expression is reduced in cystic fibrosis murine and human airway epithelial cells. J Clin Invest102, 1200-1207.[CrossRef][Google Scholar]
Kitzler, J. W., Minakami, H. & Fridovich, I.(1990). Effects of Paraquat on Escherichia coli: differences between B and K-12 strains. J Bacteriol172, 686-690.
[Google Scholar]
Koch, C. & Hoiby, N.(1993). Pathogenesis of cystic fibrosis. Lancet341, 1065-1069.[CrossRef][Google Scholar]
LiPuma, J. J., Dasen, S. E., Nielson, D. W., Stern, R. C. & Stull, T. L.(1990). Person-to-person transmission of Pseudomonas cepacia between patients with cystic fibrosis. Lancet336, 1094-1096.[CrossRef][Google Scholar]
Loewen, P. C. & Triggs, B. L.(1984). Genetic mapping of katF, a locus that with katE affects the synthesis of a second catalase species in Escherichia coli. J Bacteriol160, 668-675.
[Google Scholar]
Mahenthiralingam, E., Campbell, M. E., Henry, D. A. & Speert, D.(1996). Epidemiology of Burkholderia cepacia infection in patients with cystic fibrosis: analysis by randomly amplified polymorphic DNA fingerprinting. J Clin Microbiol34, 2914-2930.
[Google Scholar]
Manca, C., Paul, S., Barry, C. L. I., Freedman, V. H. & Kaplan, G.(1999).Mycobacterium tuberculosis catalase and peroxidase activities and resistance to oxidative killing in human monocytes in vitro. Infect Immun67, 74-79.
[Google Scholar]
Mandell, G. L.(1975). Catalase, superoxide dismutase, and virulence of Staphylococcus aureus. In vitro and in vivo studies with emphasis on staphylococcal–leukocyte interaction. J Clin Invest55, 561-566.[CrossRef][Google Scholar]
Marcinkeviciene, J. A., Magliozzo, R. S. & Blanchard, J. S.(1995). Purification and characterization of the Mycobacterium smegmatis catalase-peroxidase involved in isoniazid activation. J Biol Chem270, 22290-22295.[CrossRef][Google Scholar]
Marolda, C. L., Hauröder, B., John, M. A., Michel, R. & Valvano, M. A.(1999). Intracellular survival and saprophytic growth of isolates from the Burkholderia cepacia complex in free-living amoebae. Microbiology145, 1509-1517.[CrossRef][Google Scholar]
Martin, D. W. & Mohr, C. D.(2000). Invasion and intracellular survival of Burkholderia cepacia. Infect Immun68, 24-29.[CrossRef][Google Scholar]
Membrillo-Hernandez, J., Coopamah, M. D., Anjum, M. F., Stevanin, T. M., Kelly, A., Hughes, M. N. & Poole, R. K.(1999). The flavohemoglobin of Escherichia coli confers resistance to a nitrosating agent, a nitric oxide releaser, and paraquat and is essential for transcriptional responses to oxidative stress. J Biol Chem274, 748-754.[CrossRef][Google Scholar]
Mongkolsuk, S., Loprasert, S., Vattanaviboon, P., Chanvanichayachai, C., Chamnongpol, S. & Supsamran, N.(1996). Heterologous growth phase- and temperature-dependent expression and H2O2 toxicity protection of a superoxide-inducible monofunctional catalase gene from Xanthomonas oryzae pv. oryzae. J Bacteriol178, 3578-3584.
[Google Scholar]
Odell, E. W. & Segal, A. W.(1991). Killing of pathogens associated with chronic granulomatous disease by the non-oxidative microbicidal mechanisms of human neutrophils. J Med Microbiol34, 129-135.[CrossRef][Google Scholar]
Palleroni, N. J.(1992). Human and animal pathogenic pseudomonads. In The Prokaryotes: a Handbook on the Biology of Bacteria; Ecophysiology, Isolation, Identification, Applications , pp. 3086-3103. Edited by A. Balows, H. G. Trüper, M. Dworkin, W. Harder & K. H. Schleifer. New York:Springer.
Saini, L., Galsworthy, S., John, M. & Valvano, M. A.(1999). Intracellular survival of isolates from the Burkholderia cepacia complex in a murine macrophage cell line. Microbiology145, 3465-3475.
[Google Scholar]
Schnell, S. & Steinman, H. M.(1995). Function and stationary-phase induction of periplasmic copper-zinc superoxide dismutase and catalase/peroxidase in Caulobacter crescentus. J Bacteriol177, 5924-5929.
[Google Scholar]
Smith, A. W., Green, J., Eden, C. E. & Watson, M. L.(1999). Nitric oxide-induced potentiation of the killing of Burkholderia cepacia by reactive oxygen species: implications for cystic fibrosis. J Med Microbiol48, 419-423.[CrossRef][Google Scholar]
Speert, D. P., Bond, M., Woodman, R. C. & Curnutte, J. T.(1994). Infection with Pseudomonas cepacia in chronic granulomatous disease: role of non-oxidative killing by neutrophils in host defense. J Infect Dis170, 1524-1531.[CrossRef][Google Scholar]
Steinman, H. M.(1993). Function of periplasmic copper-zinc superoxide dismutase in Caulobacter crescentus. J Bacteriol175, 1198-1202.
[Google Scholar]
Suttorp, N. & Simon, L. M.(1982). Lung cell oxidant injury. Enhancement of polymorphonuclear leukocyte-mediated cytotoxicity in lung cells exposed to sustained in vitro hyperoxia. J Clin Invest70, 342-350.[CrossRef][Google Scholar]
Tablan, O. C., Chroba, T. L., Schidlow, D. V. & 7 other authors (1985).Pseudomonas cepacia colonization in patients with cystic fibrosis: risk factors and clinical outcome. J Pediatr107, 382–387.[CrossRef][Google Scholar]
Tager, A. M., Wu, J. & Vermeulen, M. W.(1998). The effect of chloride concentration on human neutrophil functions: potential relevance to cystic fibrosis. Am J Respir Cell Mol Biol19, 642-652.
[Google Scholar]
Tsolis, R. M., Baumler, A. J. & Heffron, F.(1995). Role of Salmonella typhimurium Mn-superoxide dismutase (SodA) in protection against early killing by J774 macrophages. Infect Immun63, 1739-1744.
[Google Scholar]
Vandamme, P., Holmes, B., Vancanneyt, M. & 8 other authors (1997). Occurrence of multiple genomovars of Burkholderia cepacia in cystic fibrosis patients and proposal of Burkholderia multivorans sp. nov. Int J Syst Bacteriol47, 1188–1200.[CrossRef][Google Scholar]
Vandamme, P., Mahenthiralingam, E., Holmes, B., Coyene, T., Hoste, B., De Vos, P., Henry, D. & Speert, D. P.(2000). Identification and population structure of Burkholderia stabilis sp. nov. (formerly Burkholderia cepacia Genomovar IV). J Clin Microbiol38, 1042-1047.
[Google Scholar]
Wayne, L. G. & Diaz, G. A.(1986). A double staining method for differentiating between two classes of mycobacterial catalase in polyacrylamide electrophoresis gels. Anal Biochem157, 89-92.[CrossRef][Google Scholar]
Weisiger, R. A. & Fridovich, I.(1973). Superoxide dismutase: organelle specificity. J Biol Chem248, 3582-3592.
[Google Scholar]
Welch, D. F., Sword, C. P., Brehm, S. & Dusanic, D.(1979). Relationship between superoxide dismutase and pathogenic mechanisms of Listeria monocytogenes. Infect Immun23, 863-872.
[Google Scholar]
Wilks, K. E., Dunn, K. L. R., Farrant, J. L., Reddin, K. M., Gorringe, A. R., Langford, P. R. & Kroll, J. S.(1998). Periplasmic superoxide dismutase in meningococcal pathogenicity. Infect Immun66, 213-217.
[Google Scholar]
Worthen, G. S., Seccombe, J. F., Clay, K. L., Guthrie, L. A. & Johnston, R. B. J.(1988). The priming of neutrophils by lipopolysaccharide for production of intracellular platelet-activating factor. Potential role in mediation of enhanced superoxide secretion. J Immunol140, 3553-3559.
[Google Scholar]
Xiu, Q. & Pan, S. Q.(2000). An Agrobacterium catalase is a virulence factor involved in tumourigenesis. Mol Microbiol35, 407-414.[CrossRef][Google Scholar]
Zughaier, S. M., Ryley, H. C. & Jackson, S. K.(1999). Lipopolysaccharide (LPS) from Burkholderia cepacia is more active than LPS from Pseudomonas aeruginosa and Stenotrophomonas maltophilia in stimulating tumour necrosis factor alpha from human monocytes. Infect Immun67, 1505-1507.
[Google Scholar]