Restorative Dentistry and Endodontics

The Korean Academy of Conservative Dentistry (대한치과보존학회)
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A STUDY ON THE HEMOLYTIC PROPERTIES OF PREVOTELLA NIGRESCENS

Prevotella nigrescens의 용혈특성에 관한 연구

  • Kwak, Ju-Seok (Department of Conservative Dentistry, College of Dentistry, Wonkwang University) ;
  • Jang, Hoon-Sang (Department of Conservative Dentistry, College of Dentistry, Wonkwang University) ;
  • Jang, Seok-Woo (Department of Conservative Dentistry, College of Dentistry, Wonkwang University) ;
  • Lee, Su-Jong (Department of Conservative Dentistry, College of Dentistry, Wonkwang University) ;
  • Yu, Yong-Wook (Department of Oral Biochemistry, College of Dentistry, Wonkwang university) ;
  • Min, Kyung-San (Department of Conservative Dentistry, College of Dentistry, Wonkwang University)
  • 곽주석 (원광대학교 치과대학 치과보존학교실) ;
  • 장훈상 (원광대학교 치과대학 치과보존학교실) ;
  • 장석우 (원광대학교 치과대학 치과보존학교실) ;
  • 이수종 (원광대학교 치과대학 치과보존학교실) ;
  • 유용욱 (원광대학교 치과대학 구강생화학교실) ;
  • 민경산 (원광대학교 치과대학 치과보존학교실)
  • Published : 2005.07.01
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Abstract

Hemolytic property is a specific feature of bacteria to obtain iron which is essential for its survival in host tissues. Therefore, it is thought to be one of several factors of virulence. The purpose of this study was to investigate the hemolytic properties of Prevotella nigrescens isolated from the teeth diagnosed as pulp necrosis and apical periodontitis under the presence of hemolysin inhibitors such as $NaN_3$ and dithiothreitol. heat, various pH and cultural conditions. The results were as follows; 1. Clinically isolated P. nigrescens strains and standard P. nigrscens ATCC 33563 showed hemolytic activity. 2. P. nigrescens showed higher hemolytic activity against human erythrocytes than sheep or horse erythrocytes. 3. $NaN_3$ and dithiothreitol (DTT) reduced the hemolytic activity of P. nigrescens in a dose dependent manner (p<0.05). 4. Optimal pH for the maximum hemolytic activity of P. nigrescens was 4.0 and the hemolysin was stable under the $50^{\circ}C$, but the hemolytic activity was significantly decreased at $95^{\circ}C$. 5. P. nigrescens cultured in $10\%\;CO_2$ condition showed higher hemolytic activity than the bacteria cultured in the anaerobic condition.

세균의 용혈활성은 세균이 숙주 내에서 생존하기 위해 필요한 철을 획득하기 위한 특성이며 기능 면에서 볼 때 숙주에 대한 중요한 독력인자로 간주될 수 있다. 본 연구에서는 괴사치수 및 치근단 치주염으로 진단된 환자의 근관에서 분리한 Prevotella nigrescens의 용혈활성을 다양한 조건 하에서 측정하여 그 특성을 규명하는 것을 목적으로 한다. 1. 임상에서 분리한 P. nigrescens와 표준균주인 P. nigrescens ATCC 33563에서 모두 용혈활성이 나타났다. 2. 사람, 면양 및 말 세 가지 종에 대해 용혈활성을 비교한 결과 사람의 적혈구에서 가장 강한 용혈활성을 나타내었다. 3. 용혈소 억제제인 $NaN_3$와 dithiothreitol(DTT)는 농도의존적으로 P. nigrescens의 용혈활성을 감소시켰다 (p<0.05) 4. P. nigrescens가 최대 용혈활성을 나타내는 최적의 pH는 4이었으며, $50^{\circ}C$이하의 온도에서는 용혈활성을 보였으나 $95^{\circ}C$에서 급격히 감소하였다. 5.배양조건에 따른 P. nigrescens의 용혈활성을 비교한 결과 $10\%\;CO_2$배양기에 배양한 경우 혐기성 조건에서 배양한 것보다 더 높은 용혈활성을 보였다.

Keywords

References

  1. Paster BJ, Boches SK, Galvin JL, Ericson RE, Lau CN, Levanos VA, Sahasrabudhe A, Dewhirst FE. Bacterial diversity in human subgingival plaque. J Bacteriol 183:3770-3783, 2001 https://doi.org/10.1128/JB.183.12.3770-3783.2001
  2. Slots J, Kamma JJ. General health risk of periodontal disease. Int Dent J 51:417-427, 2001 https://doi.org/10.1002/j.1875-595X.2001.tb00854.x
  3. Loesche WJ. Role of anaerobic bacteria in periodontal disease. Ann Otol Rhinol Laryngol Suppl 154:43-45, 1991
  4. Daly CG, Seymour GJ, Kieser JB. Bacterial endotoxin: a role in chronic inflammatory periodontal disease? J Oral Pathol 9:1-15, 1980 https://doi.org/10.1111/j.1600-0714.1980.tb01383.x
  5. Daly C, Fitzgerald GF, Davis R. Biotechnology of lactic acid bacteria with special reference to bacteriophage resistance. Antonie Van Leeuwenhoek 70:99-110, 1996 https://doi.org/10.1007/BF00395928
  6. Daly C, Mitchell D, Grossberg D, Highfield J, Stewart D. Bacteraemia caused by periodontal probing. Aust Dent J 42:77-80, 1997 https://doi.org/10.1111/j.1834-7819.1997.tb00100.x
  7. Shah HN, Gharbia SE. Biochemical and chemical studies on strains designated Prevotella intermedia and proposal of a new pigmented species. Prevotella nigrescens sp. nov. Int J Syst Bacteriol 42:542-546, 1992 https://doi.org/10.1099/00207713-42-4-542
  8. Takahashi N, Yamada T. Glucose metabolism by Prevotella intermedia and Prevotella nigrescens. Oral Microbiol Immunol 15:188-195, 2000 https://doi.org/10.1034/j.1399-302x.2000.150307.x
  9. Amano A, Kuboniwa M, Kataoka K, Tazaki K, Inoshita E, Nagata H, Tamagawa H, Shizukuishi S. Binding of hemoglobin by Porphyromonas gingivalis. FEMS Microbiol Lett 134:63-67, 1995 https://doi.org/10.1111/j.1574-6968.1995.tb07915.x
  10. Chattopadhyay K, Bhattacharyya D, Banerjee KK. Vibrio cholerae hemolysin. Eur J Biochem 269:4351, 2002 https://doi.org/10.1046/j.1432-1033.2002.03137.x
  11. Eberhard TH, Sledjeski DD, Boyle MD. Mouse skin passage of a Streptococcus pyogenes Tn917 mutant of sagA/pel restores virulence, beta-hemolysis and sagN/pel expression without altering the position or sequence of the transposon. BMC Microbiol 1:33, 2001 https://doi.org/10.1186/1471-2180-1-33
  12. Jurgens D, Ozel M, Takaisi-Kikuni NB. Production and characterization of Escherichia coli enterohemolysin and its effects on the structure of erythrocyte membranes. Cell Biol Int 26:175-186, 2002 https://doi.org/10.1006/cbir.2001.0831
  13. Hertle R, Hilger M, Weingardt-Kocher S, Walev I. Cytotoxic action of Serratia marcescens hemolysin on human epithelial cells. Infect Immun 67:817-825, 1999
  14. Ali-Vehmas T, Vikerpuur M, Pyorala S, Atroshi F. Characterization of hemolytic activity of Staphylococcus aureus strains isolated from bovine mastitic milk. Microbiol Res 155:339-344, 2001 https://doi.org/10.1016/S0944-5013(01)80013-6
  15. Byrd W, Hooke AM. Immunization with temperature-sensitive mutants of Actinobacillus pleuropneumoniae induces protective hemolysin-neutralizing antibodies in mice. Curr Microbiol 34:149-154, 1997 https://doi.org/10.1007/s002849900160
  16. Chu L, Bramanti TE, Ebersole JL, Holt SC. Hemolytic activity in the periodontopathogen Porphyromonas gingivalis: kinetics of enzyme release and localization. Infect Immun 59:1932-1940, 1991
  17. Haque A, Sugimoto N, Horiguchi Y, Okabe T, Miyata T, Iwanaga S, Matsuda M. Production, purification, and characterization of botulinolysin, a thiol-activated hemolysin of Clostridium botulinum. Infect Immun 60:71-78, 1992
  18. Glomski IJ, Gedde MM, Tsang AW, Swanson JA, Portnoy DA. The Listeria monocytogenes hemolysin has an acidic pH optimum to compartmentalize activity and prevent damage to infected host cells. J Cell Biol 156:1029-1038, 2002 https://doi.org/10.1083/jcb.200201081
  19. Falkler WA Jr, Clayman EB, Shaefer DF. Haemolysis of human erythrocytes by the Fusobacterium nucleatum associated with periodontal disease. Arch Oral Biol 28:735-739, 1983 https://doi.org/10.1016/0003-9969(83)90109-7
  20. Okamoto M, Maeda N, Kondo K, Leung KP. Hemolytic and hemagglutinating activities of Prevotella intermedia and Prevotella nigrescens. FEMS Microbiol Lett 178:299-304, 1999 https://doi.org/10.1111/j.1574-6968.1999.tb08691.x
  21. Sayers NM, James JA, Drucker DB, Blinkhorn AS. Possible potentiation of toxins from Prevotella in tennedia, Prevotella nigrescens, and Porphyromonas gingivalis by cotinin. J Periodontol 70:1269-1275, 1999 https://doi.org/10.1902/jop.1999.70.11.1269
  22. Gharbia SE, Haapasalo M, Shah HN, Kotiranta A, Lounatmaa K, Pearce MA, Devine DA. Characterization of Prevotella intermedia and Prevotella nigrescens isolates from periodontic and endodontic infections. J Periodontol 65:56-61, 1994 https://doi.org/10.1902/jop.1994.65.1.56
  23. Bae KS, Baumgartner JC, Shearer TR, David LL. Occurrence of Prevotella nigrescens and Prevotella intermedia in infections of endodontic origin. J Endod 23:620-623, 1997 https://doi.org/10.1016/S0099-2399(97)80173-2
  24. Scheffer J, Konig W, Braun V, Goebel W. Comparison of four hemolysin-producing organisms (Escherichia coli, Serratia marcescens, Aeromonas hydrophila, and Listeria monocytogenes) for release of inflammatory mediators from various cells. J Clin Microbiol 26:544-551, 1988
  25. Beem JE, Nesbitt WE, Leung KP. Identification of hemolytic activity in Prevotella intermedia. Oral Microbiol Immunol 13:97-105, 1998 https://doi.org/10.1111/j.1399-302X.1998.tb00719.x
  26. Gadeberg OV, Orskov I. In vitro cytotoxic effect of alphahemolytic Escherichia coli on human blood granulocytes. Infect Immun 45:255-260, 1984
  27. Konig B, Konig W. Effect of growth factors on Escherichia coli alpha-hemolysin-induced mediator release from human inflammatory cells: involvement of the signal transduction pathway. Infect Immun 62:2085-2093, 1994
  28. Krasilnikov OV, Merzlyak PG, Yuldasheva LN, Rodrigues CG, Bhakdi S, Valeva A. Electrophysiological evidence for heptameric stoichiometry of ion channels formed by Staphylococcus aureus alpha-toxin in planar lipid bilayers. Mol Microbiol 37:1372-1378, 2000 https://doi.org/10.1046/j.1365-2958.2000.02080.x
  29. Mengaud J, Chenevert J, Geoffroy C, Gaillard JL, Cossart P. Identification of the structural gene encoding the SH-activated hemolysin of Listeria monocytogenes: listeriolysin O is homologous to streptolysin O and pneumolysin. Infect Immun 55:3225-3227, 1987