Table of Contents  
ORIGINAL ARTICLE
Year : 2016  |  Volume : 8  |  Issue : 2  |  Page : 92-95

Detection and prevalence of Capnocytophaga in periodontal Health and disease


Department of Periodontology, Maratha Mandal's Nathajirao G. Halgekar Institute of Dental Science's and Research Center, Belgaum, Karnataka, India

Date of Web Publication16-Dec-2016

Correspondence Address:
Abhinav S Baheti
Dwarka Maternity Home, Junapress, Tal at Post Shevgaon, Ahmednagar - 414 502, Maharashtra
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/0975-8844.195911

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  Abstract 

Context/Background: Periodontal disease is a multifactorial disease, in which bacteria play a major role. Capnocytophaga species form a part of human oral flora both in health and disease. They have been implicated as putative periodontal pathogens, and yet, they are less understood members of plaque flora. No studies have been conducted on the association of Capnocytophaga species with periodontal diseases in India. Aim: The aim of this study was to detect the prevalence of Capnocytophaga species in patients with healthy periodontium, gingivitis, and periodontitis using culture method. Methods: Forty patients each with healthy periodontium, gingivitis, and periodontitis were selected. Subgingival plaque samples were collected from all the patients using sterile curettes and transferred to transport medium and sent to the laboratory. The plaque samples were inoculated on blood agar and trypticase-blood-bacitracin-polymyxin agar to grow Capnocytophaga species. Later, Gram-staining and microscopy were done to confirm the presence of Capnocytophaga in each sample. The prevalence of Capnocytophaga species was statistically analyzed using Chi-square test, Kruskal–Wallis analysis of variance, and Mann–Whitney U-test. Results: Capnocytophaga was detected in 21 (52.50%) samples out of 40 samples of gingivitis group, 11 (27.50%) samples of healthy group, and 12 (30%) samples of periodontitis group. Conclusions: Capnocytophaga is more prevalent in gingivitis compared to healthy periodontium and periodontitis. Capnocytophaga has the potential to cause periodontal disease, but as it is less competitive in the periodontal pocket, it is usually overgrown by other rapidly growing bacteria.

Keywords: Capnocytophaga, culture, gingivitis, periodontitis


How to cite this article:
Pudakalkatti PS, Baheti AS, Hattarki SA, Kambali SS, Naik RM. Detection and prevalence of Capnocytophaga in periodontal Health and disease. J Orofac Sci 2016;8:92-5

How to cite this URL:
Pudakalkatti PS, Baheti AS, Hattarki SA, Kambali SS, Naik RM. Detection and prevalence of Capnocytophaga in periodontal Health and disease. J Orofac Sci [serial online] 2016 [cited 2017 Apr 28];8:92-5. Available from: http://www.jofs.in/text.asp?2016/8/2/92/195911


  Introduction Top


Capnocytophaga species consist of Gram-negative rods, most of which are residents of normal oral microflora. “Capnos” means “smoke or carbon dioxide (CO2),” whereas “cytophaga” means “cell digester.” Capnocytophaga species typically are aerotolerant anaerobes which require significant quantities of CO2 for growth under aerobic as well as anaerobic conditions and with the ability to form thin, spreading colonies on solid media.[1],[2],[3]

An early study by Newman et al. has implicated them as a periodontal pathogen,[4] whereas a study by Dzink et al. questioned this association and stated that Capnocytophaga ochracea is a beneficial species.[5] The Capnocytophaga species form a part of the human oral flora both in health and disease.[6] However, their role, whether they are commensal of the oral cavity or pathogens responsible for the periodontal disease, is not clear. They have been suggested as putative periodontal pathogens,[7] and yet, they are less studied and understood members of plaque flora.

Aim

The aim of this study was to detect the prevalence of Capnocytophaga species in patients with healthy periodontium, gingivitis, and periodontitis using culture method.


  Materials and Methods Top


The cases for the study were selected from the patients visiting the Department of Periodontology from April 2012 to October 2012, and the samples were analyzed at the Department of Molecular Biology and Immunology. The study cases were taken from the patient pool, visiting the Department of Periodontology to provide a more representative study sample. The study was approved by the Ethical Committee of the institute. The study cases were explained about the study and written informed consent was obtained from all the participants.

Healthy, gingivitis, and periodontitis groups were made,[8] and forty patients were included in each group by simple random sampling. Inclusion criteria for healthy group were: absence of any clinical sign of gingival inflammation, probing depth ≤3 mm, and no clinical attachment loss. Inclusion criteria for gingivitis group were: generalized presence of clinical signs of gingival inflammation, probing depth ≤3 mm, and no clinical attachment loss. Whereas inclusion criteria for periodontitis group were: generalized presence of clinical signs of gingival inflammation, generalized probing depth ≥5 mm, and generalized clinical attachment loss of ≥3 mm.

Exclusion criteria were: patients with any systemic disease, smokers, pregnant or lactating women, cervical/proximal/subgingival caries or restorations, periodontal or antimicrobial therapy within 3 months before sampling.

Plaque samples were collected from subgingival sites from the base of gingival sulcus or pocket in healthy, gingivitis, and periodontitis groups using sterile curettes. Sample from each subject was immediately transferred to the reduced transport fluid (transport medium) and was sent to the laboratory of the Department of Molecular Biology and Immunology. The prevalence of Capnocytophaga species in healthy periodontium, gingivitis, and periodontitis patients was detected using culture technique. The plaque samples were inoculated on blood agar and trypticase-blood-bacitracin-polymyxin (selective medium for Capnocytophaga) agar [Figure 1]. Then, plates were inoculated for 24–72 hin the presence of 10% CO2. Characteristic gliding motility on blood agar was used to identify the growth of Capnocytophaga [Figure 2]. Later, Gram-staining and microscopy were done to confirm the presence of Capnocytophaga in each sample [Figure 3].
Figure 1: Blood agar and trypticase-blood-bacitracin-polymyxin agar showing Capnocytophaga colonies

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Figure 2: Blood agar showing gliding motility of Capnocytophaga

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Figure 3: Gram-staining showing Gram-negative fusiform Capnocytophaga bacilli

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Statistical analysis

The prevalence of Capnocytophaga species was compared using Chi-square test, Kruskal–Wallis analysis of variance, and Mann–Whitney U-test.


  Results Top


Results of assessment of the prevalence of Capnocytophaga in forty plaque samples from each group are depicted in [Table 1]. We found Capnocytophaga in 21 (52.50%) samples out of 40 samples of gingivitis group. In healthy group 11 (27.50%) and periodontitis group 12 (30%) samples showed the presence of Capnocytophaga. Significantly higher prevalence of Capnocytophaga was observed in gingivitis group compared to other two groups.
Table 1: Comparison of the prevalence of Capnocytophaga among three groups

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Significantly higher prevalence of Capnocytophaga was observed in gingivitis group compared to healthy group [Table 2]. Furthermore, the prevalence of Capnocytophaga was significantly higher in gingivitis group compared to periodontitis group [Table 3]. No significant difference in the prevalence between healthy and periodontitis groups was observed [Table 4].
Table 2: Comparison of the prevalence of Capnocytophaga among healthy and gingivitis groups

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Table 3: Comparison of the prevalence of Capnocytophaga among gingivitis and adult periodontitis groups

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Table 4: Comparison of the prevalence of Capnocytophaga among healthy and adult periodontitis groups

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  Discussion Top


Periodontal disease is a multifactorial disease, characterized by interaction between pathologic bacteria and host defense mechanisms which can eventually lead to periodontal destruction and tooth loss.

No studies have been conducted on the association of Capnocytophaga species with periodontal diseases in India. Moreover, as the prevalence of microorganisms varies depending on the geographical distribution, food habits, and immune response of an individual, this study was done.[9]

In the present study, the prevalence of Capnocytophaga in gingivitis group was significantly higher compared to healthy and periodontitis groups. Concentrations and incidence of Capnocytophaga were studied by Holdeman et al. in healthy gingiva, gingivitis, and periodontitis.[10] They found higher concentrations of this organism in gingivitis, and they concluded that Capnocytophaga species are not as competitive in the periodontal pocket, and their role is overestimated in the periodontal disease. Results of the present study are in line with this study.

In a study by Irving et al., gnotobiotic rats were monoinfected with different strains of Capnocytophaga, and no plaque was allowed to form during the course of study.[11] They found that all strains caused periodontal disease. It suggests that when other competitive organisms are not present, Capnocytophaga species can cause periodontal disease.

Savitt and Socransky stated that Capnocytophaga gingivalis is associated with gingivitis.[6] Dzink et al. found that Capnocytophaga ochracea is a beneficial species.[5] These studies suggest that different Capnocytophaga species are associated with different periodontal conditions. This difference in pathogenicity may be present due to uneven distribution of virulence factors among different species.[12]

This study was done in Karnataka, and as the prevalence of microorganisms varies depending on the geographical distribution,[9] care should be taken while generalizing the results of this study.

Further studies with individual Capnocytophaga species are essential to determine the exact role of Capnocytophaga in periodontal health and disease.

One of the limitations of this study is culture method used for microbial diagnosis which is less sensitive compared to newer molecular techniques available such as polymerase chain reaction.[13] The use of these newer molecular techniques could have affected the results.


  Conclusion Top


Capnocytophaga is more prevalent in gingivitis compared to healthy periodontium and periodontitis. Pathogenicity of different strains of Capnocytophaga depends on the distribution of virulence factors among different species. Capnocytophaga has the potential to cause periodontal disease, but as it is less competitive in the periodontal pocket, it is usually overgrown by other rapidly growing bacteria.

Financial support and sponsorship

The study was funded by the Department of Periodontology and Department of Molecular Biology and Immunology at Maratha Mandal's Nathajirao G. Halgekar Institute of Dental Science's and Research Center, Belgaum.

Conflicts of interest

There are no conflicts of interest.

 
  References Top

1.
Leadbetter ER, Holt SC, Socransky SS. Capnocytophaga: New genus of gram-negative gliding bacteria. I. General characteristics, taxonomic considerations and significance. Arch Microbiol 1979;122:9-16.  Back to cited text no. 1
    
2.
Holt SC, Leadbetter ER, Socransky SS. Capnocytophaga: New genus of gram-negative gliding bacteria. II. Morphology and ultrastructure. Arch Microbiol 1979;122:17-27.  Back to cited text no. 2
    
3.
Socransky SS, Holt SC, Leadbetter ER, Tanner AC, Savitt E, Hammond BF. Capnocytophaga: New genus of gram-negative gliding bacteria. III. Physiological characterization. Arch Microbiol 1979;122:29-33.  Back to cited text no. 3
    
4.
Newman MG, Socransky SS, Savitt ED, Propas DA, Crawford A. Studies of the microbiology of periodontosis. J Periodontol 1976;47:373-9.  Back to cited text no. 4
    
5.
Dzink JL, Socransky SS, Haffajee AD. The predominant cultivable microbiota of active and inactive lesions of destructive periodontal diseases. J Clin Periodontol 1988;15:316-23.  Back to cited text no. 5
    
6.
Savitt ED, Socransky SS. Distribution of certain subgingival microbial species in selected periodontal conditions. J Periodontal Res 1984;19:111-23.  Back to cited text no. 6
    
7.
Kesic L, Milasin J, Igic M, Obradovic R. Microbial etiology of periodontal disease-Mini review. Med Biol 2008;15:1-6.  Back to cited text no. 7
    
8.
Genco RJ, Goldman HM, Cohen DW. Contemporary Periodontics. St. Louis: Mosby; 1990.  Back to cited text no. 8
    
9.
Socransky SS, Haffajee AD. Periodontal microbial ecology. Periodontol 2000 2005;38:135-87.  Back to cited text no. 9
    
10.
Holdeman LV, Moore WE, Cato EP, Burmeister JA, Palcanis KG, Ranney RR. Distribution of Capnocytophaga in periodontal microfloras. J Periodontal Res 1985;20:475-83.  Back to cited text no. 10
    
11.
Irving JT, Socransky SS, Tanner AC. Histological changes in experimental periodontal disease in rats monoinfected with gram-negative organisms. J Periodontal Res 1978;13:326-32.  Back to cited text no. 11
    
12.
Laughon BE, Syed SA, Loesche WJ. API ZYM system for identification of Bacteroides spp. Capnocytophaga spp. and spirochetes of oral origin. J Clin Microbiol 1982;15:97-102.  Back to cited text no. 12
    
13.
Eick S, Pfister W. Comparison of microbial cultivation and a commercial PCR based method for detection of periodontopathogenic species in subgingival plaque samples. J Clin Periodontol 2002;29:638-44.  Back to cited text no. 13
    


    Figures

  [Figure 1], [Figure 2], [Figure 3]
 
 
    Tables

  [Table 1], [Table 2], [Table 3], [Table 4]



 

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