Key words: infection stone, biofilm, urease, glycocalyx

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1 Key words: infection stone, biofilm, urease, glycocalyx

2

3 740 林 俊秀 Fig. 1 SEM of cut surface of urinary stones: (a) S. epidermidis biofilm embedded in apatite bladder stone (Group I), (b) K. Pneumonia and E. faecalis biofilm embedded in struvite renal stone (Group I), (c) MRSA biofilm embedded in renal stone of calcium oxalate (Group M), (d) E. faecalis and non-fermentative gramnegative rod biofilm embedded in renal stone of calcium oxalate (Group M) (a) (b) (c) (d) た もの は,腎 結 石9例,尿 管 結 石1例,膀 8例 の 計18例(37.5%)で あ った.治 療 法 別 で は 胱結石 腎,尿 管 結 石 で は 経 皮 的 腎尿 管 砕 石 術 が そ れ ぞ れ 19例,6例 と大 半 を 占 め て い た.膀 胱 結 石 で は9 例 が 経 尿 道 的 砕 石 術,5例 が膀 胱高位切 開 による 膀 胱 切 石 術 で あ っ た. 感 染 症 学雑 誌 第69巻 第6号

4 Table 1 Classification of bacteria studied according to stone culture and SEM results Table 2 Organisms isolated from stone culture

5 Table 3 Urease activity among bacteria isolated from stones Table 4 Glycocalyx (toluidine blue) producibility among bacteria isolated from stones

6 Table 5 Glycocalyx (safranine) producibility among bacteria isolated from stones Table 6 Comparison of urease and glycocalyx producibility between GPC and GNR Urease Glycocalyx (toluidine blue) Glycocalyx (safranine)

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8 8) Nemoy, N. J. & Stamy, T. A.: Surgical, bacteriological, and biochemical management of Infection stone". J. A. M. A., 215: , " ) Christensen, C. D., et al.: Adherence of slime- strains of staphylococcus epider- producing midis to smooth surfaces. Infect. Immun., 37: , ) Carroll, G. & Brennan, R. V.: The role of infection in nephrolithiasis. J. Urol., 68: 88-95, ) Westbury, E. J.: Some observations on the quantitative analysis of over 1000 urinary calculi. Brit. J. Urol., 46: , ) Nickel, J. C., et al.: An ecological study of infected urinary stone genesis in an animal model. Brit. J. Urol., 59: 21-30, ) Hirano, S., et al.: Scanning electron microscopic examinations of renal stones associated with urinary infections. Jpn. J. Urol., 29: 33-40, ) Thompson, R. B. & Stamy, T. A.: Bacteriology of infected stones. Urology, 2: , ) Mayberry-Carson, K.J., et al.: Bacterial adherence and glycocalyx formation in osteomyelitis experimentally induced with staphylococcus aureus. Infect. Immune., 43: 825-1) Charton, M., et al.: Urinary tract infection in percutaneous surgery for renal calculi. J. Urol., 833, : 15-17, ) Davenport, D. S.: Usefulness of a test for 2) Edward, M., et al.: The fate of residual fragments after extracorporeal shock wave lith- significant infections with coagulase-negative slime production as a marker for clinically otripsy monotherapy of infection stones. J. staphylococci. J. Infect. Dis., 153: , Urol., 145: 6-10, ) Griffith, D. P.: Struite stones. Kidney Int., 13: 18) Ching-Lin, Tsai., et al.: Quantitation of , glycocalyx production in coagulase-negative staphylococcus. J. Orthop. Res., 6: , ) Dajani, A. M. & Shehabi, A. A.: Bacteriology and composition of infected stone. Urol., 22: , ) Christensen, G. D.: Adherence of coagulasenegative staphylocci to plastic tissue culture 6) Nickel, J.C., et al.: Ultrastructural microbial ecology of infection-induced urinary stones. J. plates: A quantitative model for the adherence Urol., 133: , of staphylococci to medical devices. J. Clin. Microbiol., 22: , 1985.

9 Studies on Characteristics of Bacteria which cause Infection Stone Toshihide HAYASHI Okayama University, Medical School Culture of stones and electrical microscopic observation were performed with stones collected from forty-eight patients who underwent surgery for urolithiasis. Thirty-six strains of bacteria in the stones obtained from stone culture were classified as nineteen strains of bacteria deriving from primary infection stones (Group A-I) and seventeen strains of bacteria deriving from metabolic stones (Group A-M). As for their ability to produce urease and glycocalyx, they were studied in comparison with forty-nine strains of stone surface-adhering bacteria (Group B). Glycocalyx producing ability was examined by the provisional quantitative toluidine blue assay and safranine straining method. As for the electrical microscopic observation, formation of biofilm bacterium was observed in all twenty cases of primary infection stones and in thirteen cases (46.4%) of twenty-eight cases of metabolic stones. Urease producing ability per stone was 6/14 (43.9%) in Group A-I, 4/13 (30.8%) in Group A-M and, 4/32 (12.5%) in Group B. There was a significant difference (p <0.05) between Group A-I and Group B. Similarly for the glycocalyx producing ratio (toluidine blue assay), the values were 9/14 (64.3%) in Group A-I, 11/3 (84.6%) in Group A-M and 10/32 (31.3%) in Group B. As for the glycocalyx producing ratio (safranine method), the values were 10/14 (71.4%) in Group A-I, 9/13 (69.2%) in Group A-M and 9/32 (28.1%) in Group B. As for the glycocalyx producing ability, Group A-I and Group A-M both showed significantly higher production ratios compared to Group B for both toluidine blue method and safranine method. From the above reuslts, it was suggested that both in the primary and secondary infection stones, the formation or growing of stones was strongly related to the glycocalyx producing ability of the bacteria.

Fig.2. Sensitivity distribution of clinical isolates of S. epidermidis (24 strains, 106 CFU/ml) Staphylococcus aureus Staphylococcus epider- midis Ent

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