A study on the hemolytic properties of Prevotella nigrescens

Ju-Seok Kwak; Hoon-Sang Jang; Seok-Woo Jang; Su-Jong Lee; Yong-Wook Yu; Kyung-San Min

doi:10.5395/JKACD.2005.30.4.335

Articles

Page Path: HOME > Restor Dent Endod > Volume 30(4); 2005 > Article

Original Article A study on the hemolytic properties of Prevotella nigrescens: Ju-Seok Kwak¹, Hoon-Sang Jang¹, Seok-Woo Jang¹, Su-Jong Lee¹, Yong-Wook Yu², Kyung-San Min¹; 2005;30(4):-343.
DOI: https://doi.org/10.5395/JKACD.2005.30.4.335
Published online: July 30, 2005

¹Department of Conservative Dentistry, College of Dentistry, Wonkwang University, Korea.

²Department of Oral Biochemistry, College of Dentistry, Wonkwang University, Korea.

Corresponding author: Kyung-San Min. Instructor, Department of Conservative Dentistry, College of Dentistry, Wonkwang University, 344-2 Shinyong-dong, Iksan, Cheonbuk, Korea, 570-749. Tel: 82-63-850-6930, Fax: 83-63-850-1932, mksdd@wonkwang.ac.kr

• Received: January 12, 2005 • Revised: February 18, 2005 • Accepted: February 21, 2005

213 Views
0 Download

prev next

Full Article

Download PDF

Abstract
REFERENCES

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 NaN₃ 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. NaN₃ 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℃, but the hemolytic activity was significantly decreased at 95℃.

5. P. nigrescens cultured in 10% CO₂ condition showed higher hemolytic activity than the bacteria cultured in the anaerobic condition.
Keywords: P. nigrescens; Hemolytic property; Iron; Virulence; Erythrocyte; Hemolysin inhibitor

Table 1

Hemolytic activity of P. nigrescens isolates

Table 2

Hemolytic activity of P. nigrescens ATCC 33563 and P. nigrescens 2304 strains based on erythrocytes

^aRelative hemolytic activity (%)

^bND; not detected

Table 3

Effect of NaN3 on hemolytic activity of P. nigrescens ATCC 33563 and P. nigrescens 2304

^aRelative hemolytic activity (%)

^bND; not detected

^*p < 0.05 compared with control by Mann-Whitney U test

Table 4

Effect of dithiothreitol (DTT) on hemolytic activity of P. nigrescens ATCC 33563 and P. nigrescens 2304

^aRelative hemolytic activity (%)

^bND; not detected

^*p < 0.05 compared with control by Mann-Whitney U test

Table 5

Effect of pH on hemolytic activity of P. nigrescens ATCC33563 and 2304

^aND; not detected

Table 6

Effect of temperature on hemolytic activity of P. nigrescens ATCC 33563 and P. nigrescens 2304

^aND; not detected

Table 7

Effect of various culture status on hemolytic activity of P. nigrescens ATCC 33563 and P. nigrescens 2304

^aND ; not detected

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. 2001;183: 3770-3783.Article PubMed PMC PDF
2. Slots J, Kamma JJ. General health risk of periodontal disease. Int Dent J. 2001;51: 417-427.Article PubMed
3. Loesche WJ. Role of anaerobic bacteria in periodontal disease. Ann Otol Rhinol Laryngol Suppl. 1991;154: 43-45.Article PubMed PDF
4. Daly CG, Seymour GJ, Kieser JB. Bacterial endotoxin: a role in chronic inflammatory periodontal disease? J Oral Pathol. 1980;9: 1-15.Article PubMed
5. Daly C, Fitzgerald GF, Davis R. Biotechnology of lactic acid bacteria with special reference to bacteriophage resistance. Antonie Van Leeuwenhoek. 1996;70: 99-110.Article PubMed PDF
6. Daly C, Mitchell D, Grossberg D, Highfield J, Stewart D. Bacteraemia caused by periodontal probing. Aust Dent J. 1997;42: 77-80.Article PubMed
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. 1992;42: 542-546.Article PubMed
8. Takahashi N, Yamada T. Glucose metabolism by Prevotella intermedia and Prevotella nigrescens. Oral Microbiol Immunol. 2000;15: 188-195.Article PubMed PDF
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. 1995;134: 63-67.Article PubMed
10. Chattopadhyay K, Bhattacharyya D, Banerjee KK. Vibrio cholerae hemolysin. Eur J Biochem. 2002;269: 4351.Article PubMed PDF
11. Eberhard TH, Sledjeski DD, Boyle MD. Mouse skin passage of a Streptococcus pyogenes Tn917 mutant of sagA/pel restores virulence, beta-hemolysis and sagA/pel expression without altering the position or sequence of the transposon. BMC Microbiol. 2001;1: 33.Article PubMed PMC PDF
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. 2002;26: 175-186.Article PubMed
13. Hertle R, Hilger M, Weingardt-Kocher S, Walev I. Cytotoxic action of Serratia marcescens hemolysin on human epithelial cells. Infect Immun. 1999;67: 817-825.Article PubMed PMC PDF
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. 2001;155: 339-344.Article PubMed
15. Byrd W, Hooke AM. Immunization with temperature-sensitive mutants of Actinobacillus pleuropneumoniae induces protective hemolysin-neutralizing antibodies in mice. Curr Microbiol. 1997;34: 149-154.Article PubMed PDF
16. Chu L, Bramanti TE, Ebersole JL, Holt SC. Hemolytic activity in the periodontopathogen Porphyromonas gingivalis:kinetics of enzyme release and localization. Infect Immun. 1991;59: 1932-1940.Article PubMed PMC PDF
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. 1992;60: 71-78.Article PubMed PMC PDF
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. 2002;156: 1029-1038.Article PubMed PMC PDF
19. Falkler WA Jr, Clayman EB, Shaefer DF. Haemolysis of human erythrocytes by the Fusobacterium nucleatum associated with periodontal disease. Arch Oral Biol. 1983;28: 735-739.Article PubMed
20. Okamoto M, Maeda N, Kondo K, Leung KP. Hemolytic and hemagglutinating activities of Prevotella intermedia and Prevotella nigrescens. FEMS Microbiol Lett. 1999;178: 299-304.Article PubMed
21. Sayers NM, James JA, Drucker DB, Blinkhorn AS. Possible potentiation of toxins from Prevotella intermedia, Prevotella nigrescens, and Porphyromonas gingivalis by cotinin. J Periodontol. 1999;70: 1269-1275.PubMed
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. 1994;65: 56-61.Article PubMed
23. Bae KS, Baumgartner JC, Shearer TR, David LL. Occurrence of Prevotella nigrescens and Prevotella intermedia in infections of endodontic origin. J Endod. 1997;23: 620-623.Article PubMed
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. 1988;26: 544-551.Article PubMed PMC PDF
25. Beem JE, Nesbitt WE, Leung KP. Identification of hemolytic activity in Prevotella intermedia. Oral Microbiol Immunol. 1998;13: 97-105.Article PubMed
26. Gadeberg OV, Orskov I. In vitro cytotoxic effect of alpha-hemolytic Escherichia coli on human blood granulocytes. Infect Immun. 1984;45: 255-260.Article PubMed PMC PDF
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. 1994;62: 2085-2093.Article PubMed PMC PDF
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. 2000;37: 1372-1378.Article PubMed
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. 1987;55: 3225-3227.Article PubMed PMC PDF

Tables & Figures

Table 1

Hemolytic activity of P. nigrescens isolates

Table 2

Hemolytic activity of P. nigrescens ATCC 33563 and P. nigrescens 2304 strains based on erythrocytes

^aRelative hemolytic activity (%)

^bND; not detected

Table 3

Effect of NaN3 on hemolytic activity of P. nigrescens ATCC 33563 and P. nigrescens 2304

^aRelative hemolytic activity (%)

^bND; not detected

^*p < 0.05 compared with control by Mann-Whitney U test

Table 4

Effect of dithiothreitol (DTT) on hemolytic activity of P. nigrescens ATCC 33563 and P. nigrescens 2304

^aRelative hemolytic activity (%)

^bND; not detected

^*p < 0.05 compared with control by Mann-Whitney U test

Table 5

Effect of pH on hemolytic activity of P. nigrescens ATCC33563 and 2304

^aND; not detected

Table 6

Effect of temperature on hemolytic activity of P. nigrescens ATCC 33563 and P. nigrescens 2304

^aND; not detected

Table 7

Effect of various culture status on hemolytic activity of P. nigrescens ATCC 33563 and P. nigrescens 2304

^aND ; not detected

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. 2001;183: 3770-3783.Article PubMed PMC PDF
2. Slots J, Kamma JJ. General health risk of periodontal disease. Int Dent J. 2001;51: 417-427.Article PubMed
3. Loesche WJ. Role of anaerobic bacteria in periodontal disease. Ann Otol Rhinol Laryngol Suppl. 1991;154: 43-45.Article PubMed PDF
4. Daly CG, Seymour GJ, Kieser JB. Bacterial endotoxin: a role in chronic inflammatory periodontal disease? J Oral Pathol. 1980;9: 1-15.Article PubMed
5. Daly C, Fitzgerald GF, Davis R. Biotechnology of lactic acid bacteria with special reference to bacteriophage resistance. Antonie Van Leeuwenhoek. 1996;70: 99-110.Article PubMed PDF
6. Daly C, Mitchell D, Grossberg D, Highfield J, Stewart D. Bacteraemia caused by periodontal probing. Aust Dent J. 1997;42: 77-80.Article PubMed
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. 1992;42: 542-546.Article PubMed
8. Takahashi N, Yamada T. Glucose metabolism by Prevotella intermedia and Prevotella nigrescens. Oral Microbiol Immunol. 2000;15: 188-195.Article PubMed PDF
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. 1995;134: 63-67.Article PubMed
10. Chattopadhyay K, Bhattacharyya D, Banerjee KK. Vibrio cholerae hemolysin. Eur J Biochem. 2002;269: 4351.Article PubMed PDF
11. Eberhard TH, Sledjeski DD, Boyle MD. Mouse skin passage of a Streptococcus pyogenes Tn917 mutant of sagA/pel restores virulence, beta-hemolysis and sagA/pel expression without altering the position or sequence of the transposon. BMC Microbiol. 2001;1: 33.Article PubMed PMC PDF
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. 2002;26: 175-186.Article PubMed
13. Hertle R, Hilger M, Weingardt-Kocher S, Walev I. Cytotoxic action of Serratia marcescens hemolysin on human epithelial cells. Infect Immun. 1999;67: 817-825.Article PubMed PMC PDF
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. 2001;155: 339-344.Article PubMed
15. Byrd W, Hooke AM. Immunization with temperature-sensitive mutants of Actinobacillus pleuropneumoniae induces protective hemolysin-neutralizing antibodies in mice. Curr Microbiol. 1997;34: 149-154.Article PubMed PDF
16. Chu L, Bramanti TE, Ebersole JL, Holt SC. Hemolytic activity in the periodontopathogen Porphyromonas gingivalis:kinetics of enzyme release and localization. Infect Immun. 1991;59: 1932-1940.Article PubMed PMC PDF
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. 1992;60: 71-78.Article PubMed PMC PDF
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. 2002;156: 1029-1038.Article PubMed PMC PDF
19. Falkler WA Jr, Clayman EB, Shaefer DF. Haemolysis of human erythrocytes by the Fusobacterium nucleatum associated with periodontal disease. Arch Oral Biol. 1983;28: 735-739.Article PubMed
20. Okamoto M, Maeda N, Kondo K, Leung KP. Hemolytic and hemagglutinating activities of Prevotella intermedia and Prevotella nigrescens. FEMS Microbiol Lett. 1999;178: 299-304.Article PubMed
21. Sayers NM, James JA, Drucker DB, Blinkhorn AS. Possible potentiation of toxins from Prevotella intermedia, Prevotella nigrescens, and Porphyromonas gingivalis by cotinin. J Periodontol. 1999;70: 1269-1275.PubMed
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. 1994;65: 56-61.Article PubMed
23. Bae KS, Baumgartner JC, Shearer TR, David LL. Occurrence of Prevotella nigrescens and Prevotella intermedia in infections of endodontic origin. J Endod. 1997;23: 620-623.Article PubMed
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. 1988;26: 544-551.Article PubMed PMC PDF
25. Beem JE, Nesbitt WE, Leung KP. Identification of hemolytic activity in Prevotella intermedia. Oral Microbiol Immunol. 1998;13: 97-105.Article PubMed
26. Gadeberg OV, Orskov I. In vitro cytotoxic effect of alpha-hemolytic Escherichia coli on human blood granulocytes. Infect Immun. 1984;45: 255-260.Article PubMed PMC PDF
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. 1994;62: 2085-2093.Article PubMed PMC PDF
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. 2000;37: 1372-1378.Article PubMed
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. 1987;55: 3225-3227.Article PubMed PMC PDF

Citations

Citations to this article as recorded by

CanvasJS.com

ePub Link

Cite

CITE: export Copy Download Format; Close

Download Citation

Download a citation file in RIS format that can be imported by all major citation management software, including EndNote, ProCite, RefWorks, and Reference Manager.

Format:

RIS — For EndNote, ProCite, RefWorks, and most other reference management software
BibTeX — For JabRef, BibDesk, and other BibTeX-specific software

Include:

Citation for the content below
Citation and abstract for the content below

A study on the hemolytic properties of Prevotella nigrescens

J Korean Acad Conserv Dent. 2005;30(4):335-343. Published online July 30, 2005

DOI: https://doi.org/10.5395/JKACD.2005.30.4.335

XML Download

A study on the hemolytic properties of Prevotella nigrescens

Table 1

Hemolytic activity of P. nigrescens isolates

Table 2

Hemolytic activity of P. nigrescens ATCC 33563 and P. nigrescens 2304 strains based on erythrocytes

^aRelative hemolytic activity (%)

^bND; not detected

Table 3

Effect of NaN3 on hemolytic activity of P. nigrescens ATCC 33563 and P. nigrescens 2304

^aRelative hemolytic activity (%)

^bND; not detected

^*p < 0.05 compared with control by Mann-Whitney U test

Table 4

Effect of dithiothreitol (DTT) on hemolytic activity of P. nigrescens ATCC 33563 and P. nigrescens 2304

^aRelative hemolytic activity (%)

^bND; not detected

^*p < 0.05 compared with control by Mann-Whitney U test

Table 5

Effect of pH on hemolytic activity of P. nigrescens ATCC33563 and 2304

^aND; not detected

Table 6

Effect of temperature on hemolytic activity of P. nigrescens ATCC 33563 and P. nigrescens 2304

^aND; not detected

Table 7

Effect of various culture status on hemolytic activity of P. nigrescens ATCC 33563 and P. nigrescens 2304

^aND ; not detected

Table 1 Hemolytic activity of P. nigrescens isolates

Table 2 Hemolytic activity of P. nigrescens ATCC 33563 and P. nigrescens 2304 strains based on erythrocytes

^aRelative hemolytic activity (%)

^bND; not detected

Table 3 Effect of NaN3 on hemolytic activity of P. nigrescens ATCC 33563 and P. nigrescens 2304

^aRelative hemolytic activity (%)

^bND; not detected

^*p < 0.05 compared with control by Mann-Whitney U test

Table 4 Effect of dithiothreitol (DTT) on hemolytic activity of P. nigrescens ATCC 33563 and P. nigrescens 2304

^aRelative hemolytic activity (%)

^bND; not detected

^*p < 0.05 compared with control by Mann-Whitney U test

Table 5 Effect of pH on hemolytic activity of P. nigrescens ATCC33563 and 2304

^aND; not detected

Table 6 Effect of temperature on hemolytic activity of P. nigrescens ATCC 33563 and P. nigrescens 2304

^aND; not detected

Table 7 Effect of various culture status on hemolytic activity of P. nigrescens ATCC 33563 and P. nigrescens 2304