Key words: change serotype, Pseudomonas aeruginosa anti-pseudomonal drug,

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1 Key words: change serotype, Pseudomonas aeruginosa anti-pseudomonal drug,

2 Table 1 Incidence of changes in serotype of Pseudomonas aeruginosa isolates induced by anti-pseudomonal drugs Number of isolates changed in serotype Number of isolates tested *1); 1-4MIC, 35t, 18-hr incubation *2); Test isolates

3 Table 2 Changes in serotype of Pseudomonas aeruginosa isolates induced by anti-pseudomonal drugs *1); No change *2); Non -typable Table 3 Effect of drug concentrations on change in serotype of Pseudomonas aeruginosa isolates induced by anti-pseudomonal drugs Drug concentration-serotype Incubation; 35 Ž, 18hours *; No change

4 Fig. 1 Growth curves of serotype variants of Pseudomonas aeruginosa TA-2 induced by cefsulodin. a) Control (Drug free) b) CFS 1/2MIC (0.78pg/ml) c) CFS 2NIIC (3.13ug/ml) Fig. 2 Growth curves of serotype variants of Pseudomonas aeruginosa TA-13 induced by gentamicin. a) Control (Drug free) b) GM 1/2MIC (6.25}ig/ml)

5 Fig. 3 Changes in drug-susceptibility of serotype variants of Pseudomonas aeruginosa TA-13 induced by anti-pseudomonal drugs. (Variant MIC/parent MIC)

6 Fig. 4 Changes in drug-susceptibility of serotype variants of Pseudomonas aeruginosa TA-2 induced by anti-pseudomonal drugs. (Variant MIC/parent MIC)

7 3) Ojeniyi, B.,Rosdahl, V. T. & Hoiby, N.: Changes in serotype caused by cell to cell contact between different Pseudomonas aeruginosa strains from cystic fibrosis patients. Acta Path. Microbiol. Immunol. Scand. B, 95: 23-27, ) Ojeniyi, B.: Bacteriophages in sputum of cystic fibrosis patients as a possible cause of in vivo changes in serotypes of Pseudomonas aeruginosa. Acta Path. Microbiol. Immunol. Scand., 96: , ) Lennette, E. H., Balows, A., Hausler, W. J. Jr. & Truant, J. P.: Manual of Clinical Microbiol- 3rd ed., American Society for ogy, Microbiol- ogy, Washington, D. C., Pseudomonas, p , ) Homma, J. Y., Kim, K. S., Yamada, H., Ito, M., Shionoya, H. & Kawabe, Y.: Serological typing of Pseudomonas aeruginosa and its cross- infection. Jpn. J. Exp. Med., 40: , ) Pitt,T. L.: Epidemiological typing Pseudomonas aeruginosa. Eur. J. Clin. Microbiol. Infect. Dis., 7: , ) Lanyi, B. & Lantos, J.: Antigenic changes in P.aeruginosa in vivo and after lysogenisation in vitro. Acta Microbiol. Acad. Sci. Hung., 23: , ) Tejedor, C., Foulds, J. & Zasloff, M.: Bacteriophages in sputum of patients with bron-

8 chopulmonary Pseudomonas infection. Infec. Immunity36: , ) Ojeniyi, B., Wolz, C., Doring, G., Lam, J. S., Rosdahl, V. T. & Hoiby, N.: Typing of polyagglutinable Pseudomonas aeruginosa isolates from cystic fibrosis patients. Acta Path. Microbiol. Immunol. Scand., 98: , ) Kropinski, A. M., Auzio, J., Angus, B. L. & Hancock, R. E. W.: Chemical and chromatographic analysis of lipopolysaccharide from an antibiotic-supersusceptible mutant of Pseudomonas aeruginosa. Antimicrob. Agents Chemother., 21; , ) Legakis, N. J., Aliferopoulou, M. & Papapetropoulou, M.: Serotypes of Pseudomonas aeruginosa in clinical specimens in relation to antibiotic susceptibility. J. Clin. Microb., 16: , ) Livermore, D. M.: Penicillin-binding proteins, porins and outer-membrane permeability of carbenicillin-resistant and -susceptible strains of Pseudomonas aeruginosa. J. Med. Microb., 18: , ) Livermore, D. M. & Pitt, T. L.: Dissociation of surface properties and "intrinsic" resistance to fl-lactams in Pseudomoas aeruginosa. J. Med. Microb., 22: , ) Trias, J. & Nikaido, H.: Protein D2 channel of the Pseudomonas aeruginosa outer membrane has a binding site for for basic amino acids and peptides. J. Biol. Chem., 265: , Changes in Serotypes of Clinical Isolates of Pseudomonas aeruginosa by Anti-Pseudomonal Drugs Intetsu KOBAYASHI1,2), Miyuki HASEGAWA1), Shuichi MIYAZAKP), Minoru NISHIDAL2) Sachiko GOT02) 1) Chemotherapy Division, Mitsubishi-Yuka Bio-Clinical Laboratories 2)Department of Microbiology, School of Medicine, Toho University Forty-two isolates of P. aeruginosa from various infections were each incubated in Mueller-Hinton broth including piperacillin, cefsulodin, ceftazidime, imipenem, gentamicin or norfloxacin (1MIC- 4MIC) at 35 Ž for 18 hours, and serotyped using monoclonal antibodies. Serotypes of 4 (9.5%)-8 (19.0%) of the 42 isolates each changed to different groups after incubation. No relationship was found between serotypes of the formed variants and anti-pseudomonal drugs used. When P. aeruginosa TA-2 was exposed to cefsulodin at different concentrations (1MIC and 2MIC) under the above conditions, the distinct variants different in serotypes were formed according to the drug concentrations. Furthermore, P. aeruginosa TA-2 and TA-13 were incubated in Mueller-Hinton broth including cefsulodin (1/2 or 2MIC) and gentamicin (1/2MIC), respectively, and the growth curves of parent and variant cells were determined. In the experiments, the variants appeared 6 hours after onset of the incubation and grew with the parents. The present results may explain our findings previously reported; coexistence of colonies different in serotype of P. aeruginosa isolated from some individual patients. The results indicate the possibility that alteration in bacterial surfaces in association with changes in serotypes might occur in vivo in these patients infected with P. aeruginosa and treated with anti-pseudomonal drugs.

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