Effect of Bdellovibrio bacteriovorus HD100 on multispecies oral communities.

The predation of Bdellovibrio bacteriovorus on different periodontal pathogens has already been described. However, it is necessary to consider the polymicrobial nature of periodontal disease. The current study explores the predation of Bdellovibrio on oral pathogens organized in multispecies communities. The effect of the predator was evaluated on in vitro six species communities with microbial culturing. Additionally, the effect on ex vivo subgingival plaque and saliva samples from periodontitis patients was assessed. In the latter experiment results were examined with microbial culturing, quantitative polymerase chain reaction (qPCR) and denaturing gradient gel electrophoresis (DGGE). The latter technique was used to get an overview of the whole mixed microbial population. Results showed that even in more complex models, B. bacteriovorus was still able to predate on Fusobacterium nucleatum and Aggregatibacter actinomycetemcomitans. However predation on Prevotella intermedia and Porphyromonas gingivalis could not be validated in multispecies models. The effect of Bdellovibrio was not restricted to the target bacteria. Changes in the overall ecology of the different models were evident. It could be concluded that the efficiency of predation decreased when complexity of the models increased. However, B. bacteriovorus was able to attack two important oral pathogens, F. nucleatum, and A. actinomycetemcomitans, even when present in ex vivo clinical samples. These effects still have to be validated in in vivo models to see the impact of Bdellovibrio on the whole bacterial ecology.

Citrullination and proteolytic processing of chemokines by Porphyromonas gingivalis.

The outgrowth of Porphyromonas gingivalis within the inflammatory subgingival plaque is associated with periodontitis characterized by periodontal tissue destruction, loss of alveolar bone, periodontal pocket formation, and eventually, tooth loss. Potential virulence factors of P. gingivalis are peptidylarginine deiminase (PPAD), an enzyme modifying free or peptide-bound arginine to citrulline, and the bacterial proteases referred to as gingipains (Rgp and Kgp). Chemokines attract leukocytes during inflammation. However, posttranslational modification (PTM) of chemokines by proteases or human peptidylarginine deiminases may alter their biological activities. Since chemokine processing may be important in microbial defense mechanisms, we investigated whether PTM of chemokines by P. gingivalis enzymes occurs. Upon incubation of interleukin-8 (IL-8; CXCL8) with PPAD, only minor enzymatic citrullination was detected. In contrast, Rgp rapidly cleaved CXCL8 in vitro. Subsequently, different P. gingivalis strains were incubated with the chemokine CXCL8 or CXCL10 and their PTMs were investigated. No significant CXCL8 citrullination was detected for the tested strains. Interestingly, although considerable differences in the efficiency of CXCL8 degradation were observed with full cultures of various strains, similar rates of chemokine proteolysis were exerted by cell-free culture supernatants. Sequencing of CXCL8 incubated with supernatant or bacteria showed that CXCL8 is processed into its more potent forms consisting of amino acids 6 to 77 and amino acids 9 to 77 (the 6-77 and 9-77 forms, respectively). In contrast, CXCL10 was entirely and rapidly degraded by P. gingivalis, with no transient chemokine forms being observed. In conclusion, this study demonstrates PTM of CXCL8 and CXCL10 by gingipains of P. gingivalis and that strain differences may particularly affect the activity of these bacterial membrane-associated proteases.

Inter-bacterial correlations in subgingival biofilms: a large-scale survey.

AIM: Although the complexity of the oral ecology and the ecological differences between health and disease are well accepted, a clear view on the dynamics in relation to disease is lacking. In this study, the prevalence and abundance of 20 key oral bacteria was assessed in health and disease and more importantly a closer look was given to the inter-bacterial relationships. MATERIALS AND METHODS: A blinded microbiological database was analysed in this cross-sectional, retrospective study. The database was constructed based on microbiological analyses of samples from 6308 patients, with gradations of periodontitis (healthy to periodontitis). Data concerning the abundance of 20 oral bacteria and probing pocket depth were provided. RESULTS: Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola, Eubacterium nodatum, Porphyromonas micra and Porphyromonas intermedia showed a clear increase in abundance and prevalence with increasing pocket depth. Correlation matrices illustrated that almost all microorganisms were in one way correlated to other species and most of these correlations were significant. Several beneficial bacteria showed strong correlations with other beneficial bacteria. CONCLUSION: Knowledge on bacterial correlations can pave the way for new treatment options focusing on restoring the shifted balance.

Bacterial antagonism against periodontopathogens.

BACKGROUND: The aim of the current study is to compare the prevalence of commensal bacteria, with beneficial properties, for healthy and diseased individuals and additionally to examine the inhibitory effect of some commercial dietary probiotics on periodontopathogens, comparing this inhibitory effect to that of orally derived beneficial bacteria. METHODS: Subgingival plaque samples from 35 patients (healthy and periodontitis patients) were analyzed. Growth inhibition of the periodontal pathogens Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum, and Aggregatibacter actinomycetemcomitans was examined using the agar overlay technique and agar well diffusion method. The quantification of the inhibitory effect was checked with the agar well diffusion method. RESULTS: Using the agar overlay technique, the prevalence of strains antagonistic toward P. gingivalis, A. actinomycetemcomitans, and F. nucleatum was found to be higher in healthy individuals than in individuals with periodontitis, but this could not be validated by the agar well diffusion assay. Compared with the antagonistic activity of the isolated strains, the probiotic strains overall showed a stronger inhibition of the periodontal pathogens. CONCLUSION: It was shown that some oral bacteria can cause antagonism toward periodontopathogens, and these observations underline the therapeutic potential of applications that stimulate oral health by the application of beneficial effector strains.

Do probiotics offer opportunities to manipulate the periodontal oral microbiota?

BACKGROUND: As in other fields of healthcare, probiotics have been introduced for prevention and treatment of periodontal diseases. OBJECTIVE: This review was initiated to explore whether the use of probiotics can influence the periodontal microbiota and periodontal health. MATERIALS AND METHODS: Literature on the mode of action of oral probiotics was reviewed and a systematic review was performed on the microbiological and clinical effects of oral probiotics on periodontal health. RESULTS: Three animal and 11 in vivo human studies were retrieved. Six studies reported on microbiological effects whereas eight studies report on clinical effects. Seven studies were performed on healthy or gingivitis patients and four studies on periodontitis patients. Many of the retrieved studies are pilot in nature and with low quality. The high degree of heterogeneity between studies hampered analysis. CONCLUSION: Taking into consideration all limitations, the currently available data indicate an effect of probiotics on the oral microbiota and a more limited effect on clinical periodontal outcome measures. However, there is an urgent need for properly conducted clinical trials where probiotics are used as adjuncts to standard periodontal care, similar to antibiotics, using probiotic strains with, at least at an in vitro level, proven periodontal probiotic effects.