Isolation and identification of bacteria from the root canal of the teeth diagnosed as the acute pulpitis and acute periapical abscess

Yeon-Jae Lee; Mi-Kwang Kim; Ho-Keel Hwang; Joong-Ki Kook

doi:10.5395/JKACD.2005.30.5.409

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Original Article Isolation and identification of bacteria from the root canal of the teeth diagnosed as the acute pulpitis and acute periapical abscess: Yeon-Jae Lee¹, Mi-Kwang Kim², Ho-Keel Hwang^1,3, Joong-Ki Kook^2,3; 2005;30(5):-422.
DOI: https://doi.org/10.5395/JKACD.2005.30.5.409
Published online: September 30, 2005

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

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

³Oral Biology Research Institute, College of Dentistry, Chosun University, Korea.

Corresponding author: Joong-Ki Kook. Department of Oral Biochemistry, and Oral Biology Research Institute, College of Dentistry, Chosun University, 375 Seo-suk Dong, Dong-Gu, Gwang-ju, 501-759 Korea. Tel: 82-62-230-6877, Fax: 82-62-224-3706, jkkook@chosun.ac.kr

• Received: April 11, 2005 • Revised: May 16, 2005 • Accepted: June 13, 2005

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Abstract
REFERENCES

Abstract

The aim of this study was to identify the bacteria isolated from acute endodontic lesions by cell culture and 16S rDNA sequencing. The necrotic pulpal tissue was collected from 17 infected root canals, which were diagnosed as being either an acute pulpitis or acute periapical abscess. Samples were collected aseptically from the infected pulpal tissue of the infected root canals using a barbed broach and a paper point. The cut barbed broaches and paper points were transferred to an eppendorf tube containing 500 ul of 1 X PBS. The sample solution was briefly mixed and plated onto a BHI-agar plate containing 5% sheep blood. The agar plates were incubated in a 37℃ anaerobic chamber for 7 days. The bacteria growing on the agar plate were identified by 16S rRNA coding gene (rDNA) cloning and sequencing at the species level. Among the 71 colonies grown on the agar plates, 56 strains survived and were identified. In dental caries involving the root canals, Streptococcus spp. were mainly isolated. Actinomyces, Clostridia, Bacteroides and Fusobacteria were isolated in the periapical lesion without dental caries. Interestingly, two new Actinomyces spp. (ChDC B639 and ChDC B631) were isolated in this study. These results showed that there was diversity among the species in endodontic lesions. This suggests that an endodontic infection is a mixed infection with a polymicrobial etiology. These results may offer the bacterial strains for pathogenesis studies related to an endodontic infection.
Keywords: Acute pulpitis; Acute periapical abscess; Bacteria; 16S rRNA gene

Table 1

Clinical features of the patients

Table 2

Bacteria isolated from each of patient

ChDC; Department of Oral Biochemistry, College of Dentistry, Chosun University

Table 3

Bacteria isolated from necrotized pulpal tissue

ChDC; Department of Oral Biochemistry, College of Dentistry, Chosun University

^*; GenBank accession number

Table 4

Summary of the isolated clones derived from endodontic lesions

Table 5

Comparison of the bacteria isolated from the endodontic infection according to the presence of caries in infected teeth

Table 6

Comparison of the bacteria isolated from the endodontic infection according to the presence of the apical lesion

REFERENCES

1. Gomes BP, Lilley JD, Drucker DB. Clinical significance of dental root canal microflora. J Dent. 1996;24: 47-55.Article PubMed
2. Fouad AF, Kum KY, Clawson ML, Barry J, Abenoja C, Zhu Q, Caimano M, Radolf JD. Molecular characterization of the presence of Eubacterium spp. and Streptococcus spp. in endodontic infections. Oral Microbiol Immunol. 2003;18: 249-255.Article PubMed PDF
3. Kurihara H, Kobayashi Y, Francisco IA, Isoshima O, Nagai A, Murayama YA. Microbiological and immunological study of endodontic-periodontic lesions. J Endod. 1995;21: 617-621.PubMed
4. Abou-Rass M, Bogen G. Microorganisms in closed periapical lesions. Int Endod J. 1998;31: 39-47.Article PubMed
5. Baumgartner JC, Watkins BJ, Bae KS, Xia T. Association of black-pigmented bacteria with endodontic infections. J Endod. 1999;25: 413-415.Article PubMed
6. Rolph HJ, Lennon A, Riggio MP, Saunders WP, MacKenzie D, Coldero L, Bagg J. Molecular identification of microorganisms from endodontic infections. J Clin Microbiol. 2001;39: 3282-3289.Article PubMed PMC PDF
7. Tanner A, Stillman N. Oral and dental infections with anaerobic bacteria: clinical features, predominant pathogens, and treatment. Clin Infect Dis. 1993;16: S304-S309.Article PubMed
8. Siqueira JF Jr, Rocas IN, Souto R, de Uzeda M, Colombo AP. Checkerboard DNA-DNA hybridization analysis of endodontic infections. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2000;89: 744-748.Article PubMed
9. Baumgartner JC, Bae KS, Xia T, Whitt J, David LL. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and polymerase chain reaction for differentiation of Prevotella intermedia and Prevotella nigrescens. J Endod. 1999;25: 324-328.Article PubMed
10. Siqueira JF Jr, Rocas IN, Oliveira JC, Santos KR. Molecular detection of black-pigmented bacteria in infections of endodontic origin. J Endod. 2001;27: 563-566.Article PubMed
11. Maidak BL, Olsen GJ, Larsen N, Overbeek R, McCaughey MJ, Woese CR. The ribosomal database project (RDP). Nucleic Acids Res. 1996;24: 82-85.Article PubMed PMC
12. Munson MA, Pitt-Ford T, Chong B, Weightman A, Wade WG. Molecular and cultural analysis of the microflora associated with endodontic infections. J Dent Res. 2002;81: 761-766 Erratum in: J Dent Res. 2003. 82:69. J Dent Res 2003. 82:247, 2002.Article PubMed PDF
13. Yoo SY, Kim MK, Kim HS, Hwang HK, Kim PS, Lim SY, Oh SH, Min JB, Kook JK. Identification of bacteria from periapical abscess using 16S rDNA clone libraries. Korean J Microbiol Biotechnol. 2004;32: 195-198.
14. Sundqvist G. Taxonomy, ecology, and pathogenicity of the root canal flora. Oral Surg Oral Med Oral Pathol. 1994;78: 522-530.Article PubMed
15. Le Goff A, Bunetel L, Mouton C, Bonnaure-Mallet M. Evaluation of root canal bacteria and their antimicrobial susceptibility in teeth with necrotic pulp. Oral Microbiol Immunol. 1997;12: 318-322.Article PubMed
16. Love RM, Jenkinson HF. Invasion of dentinal tubules by oral bacteria. Crit Rev Oral Biol Med. 2002;13: 171-183.PubMed
17. 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

Tables & Figures

Table 1

Clinical features of the patients

Table 2

Bacteria isolated from each of patient

ChDC; Department of Oral Biochemistry, College of Dentistry, Chosun University

Table 3

Bacteria isolated from necrotized pulpal tissue

ChDC; Department of Oral Biochemistry, College of Dentistry, Chosun University

^*; GenBank accession number

Table 4

Summary of the isolated clones derived from endodontic lesions

Table 5

Comparison of the bacteria isolated from the endodontic infection according to the presence of caries in infected teeth

Table 6

Comparison of the bacteria isolated from the endodontic infection according to the presence of the apical lesion

REFERENCES

1. Gomes BP, Lilley JD, Drucker DB. Clinical significance of dental root canal microflora. J Dent. 1996;24: 47-55.Article PubMed
2. Fouad AF, Kum KY, Clawson ML, Barry J, Abenoja C, Zhu Q, Caimano M, Radolf JD. Molecular characterization of the presence of Eubacterium spp. and Streptococcus spp. in endodontic infections. Oral Microbiol Immunol. 2003;18: 249-255.Article PubMed PDF
3. Kurihara H, Kobayashi Y, Francisco IA, Isoshima O, Nagai A, Murayama YA. Microbiological and immunological study of endodontic-periodontic lesions. J Endod. 1995;21: 617-621.PubMed
4. Abou-Rass M, Bogen G. Microorganisms in closed periapical lesions. Int Endod J. 1998;31: 39-47.Article PubMed
5. Baumgartner JC, Watkins BJ, Bae KS, Xia T. Association of black-pigmented bacteria with endodontic infections. J Endod. 1999;25: 413-415.Article PubMed
6. Rolph HJ, Lennon A, Riggio MP, Saunders WP, MacKenzie D, Coldero L, Bagg J. Molecular identification of microorganisms from endodontic infections. J Clin Microbiol. 2001;39: 3282-3289.Article PubMed PMC PDF
7. Tanner A, Stillman N. Oral and dental infections with anaerobic bacteria: clinical features, predominant pathogens, and treatment. Clin Infect Dis. 1993;16: S304-S309.Article PubMed
8. Siqueira JF Jr, Rocas IN, Souto R, de Uzeda M, Colombo AP. Checkerboard DNA-DNA hybridization analysis of endodontic infections. Oral Surg Oral Med Oral Pathol Oral Radiol Endod. 2000;89: 744-748.Article PubMed
9. Baumgartner JC, Bae KS, Xia T, Whitt J, David LL. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and polymerase chain reaction for differentiation of Prevotella intermedia and Prevotella nigrescens. J Endod. 1999;25: 324-328.Article PubMed
10. Siqueira JF Jr, Rocas IN, Oliveira JC, Santos KR. Molecular detection of black-pigmented bacteria in infections of endodontic origin. J Endod. 2001;27: 563-566.Article PubMed
11. Maidak BL, Olsen GJ, Larsen N, Overbeek R, McCaughey MJ, Woese CR. The ribosomal database project (RDP). Nucleic Acids Res. 1996;24: 82-85.Article PubMed PMC
12. Munson MA, Pitt-Ford T, Chong B, Weightman A, Wade WG. Molecular and cultural analysis of the microflora associated with endodontic infections. J Dent Res. 2002;81: 761-766 Erratum in: J Dent Res. 2003. 82:69. J Dent Res 2003. 82:247, 2002.Article PubMed PDF
13. Yoo SY, Kim MK, Kim HS, Hwang HK, Kim PS, Lim SY, Oh SH, Min JB, Kook JK. Identification of bacteria from periapical abscess using 16S rDNA clone libraries. Korean J Microbiol Biotechnol. 2004;32: 195-198.
14. Sundqvist G. Taxonomy, ecology, and pathogenicity of the root canal flora. Oral Surg Oral Med Oral Pathol. 1994;78: 522-530.Article PubMed
15. Le Goff A, Bunetel L, Mouton C, Bonnaure-Mallet M. Evaluation of root canal bacteria and their antimicrobial susceptibility in teeth with necrotic pulp. Oral Microbiol Immunol. 1997;12: 318-322.Article PubMed
16. Love RM, Jenkinson HF. Invasion of dentinal tubules by oral bacteria. Crit Rev Oral Biol Med. 2002;13: 171-183.PubMed
17. 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

Citations

Citations to this article as recorded by

Microorganism penetration in dentinal tubules of instrumented and retreated root canal walls.In vitroSEM study
Saad Al-Nazhan, Alaa Al-Sulaiman, Fellwa Al-Rasheed, Fatimah Alnajjar, Bander Al-Abdulwahab, Abdulhakeem Al-Badah
Restorative Dentistry & Endodontics.2014; 39(4): 258. CrossRef
Microbial profile of asymptomatic and symptomatic teeth with primary endodontic infections by pyrosequencing
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Journal of Korean Academy of Conservative Dentistry.2011; 36(6): 498. CrossRef

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Isolation and identification of bacteria from the root canal of the teeth diagnosed as the acute pulpitis and acute periapical abscess

J Korean Acad Conserv Dent. 2005;30(5):409-422. Published online September 30, 2005

DOI: https://doi.org/10.5395/JKACD.2005.30.5.409

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Isolation and identification of bacteria from the root canal of the teeth diagnosed as the acute pulpitis and acute periapical abscess

Table 1

Clinical features of the patients

Table 2

Bacteria isolated from each of patient

ChDC; Department of Oral Biochemistry, College of Dentistry, Chosun University

Table 3

Bacteria isolated from necrotized pulpal tissue

ChDC; Department of Oral Biochemistry, College of Dentistry, Chosun University

^*; GenBank accession number

Table 4

Summary of the isolated clones derived from endodontic lesions

Table 5

Comparison of the bacteria isolated from the endodontic infection according to the presence of caries in infected teeth

Table 6

Comparison of the bacteria isolated from the endodontic infection according to the presence of the apical lesion

Table 1 Clinical features of the patients

Table 2 Bacteria isolated from each of patient

ChDC; Department of Oral Biochemistry, College of Dentistry, Chosun University

Table 3 Bacteria isolated from necrotized pulpal tissue

ChDC; Department of Oral Biochemistry, College of Dentistry, Chosun University

^*; GenBank accession number

Table 4 Summary of the isolated clones derived from endodontic lesions

Table 5 Comparison of the bacteria isolated from the endodontic infection according to the presence of caries in infected teeth

Table 6 Comparison of the bacteria isolated from the endodontic infection according to the presence of the apical lesion