Oral Prevotella Species and Their Connection to Events of Clinical Relevance in Gastrointestinal and Respiratory Tracts

Prevotella is recognized as one of the core anaerobic genera in the oral microbiome. In addition, members of this genus belong to microbial communities of the gastrointestinal and respiratory tracts. Several novel Prevotella species, most of them of oral origin, have been described, but limited knowledge is still available of their clinical relevance. Prevotella melaninogenica is among the anaerobic commensals on oral mucosae from early months of life onward, and other early colonizing Prevotella species in the oral cavity include Prevotella nigrescens and Prevotella pallens. Oral Prevotella species get constant access to the gastrointestinal tract via saliva swallowing and to lower airways via microaspiration. At these extra-oral sites, they play a role as commensals but also as potentially harmful agents on mucosal surfaces. The aim of this narrative review is to give an updated overview on the involvement of oral Prevotella species in gastrointestinal and respiratory health and disease.

Prevotella melaninogenica, a Sentinel Species of Antibiotic Resistance in Cystic Fibrosis Respiratory Niche?

The importance and abundance of strict anaerobic bacteria in the respiratory microbiota of people with cystic fibrosis (PWCF) is now established through studies based on high-throughput sequencing or extended-culture methods. In CF respiratory niche, one of the most prevalent anaerobic genera is Prevotella, and particularly the species Prevotella melaninogenica. The objective of this study was to evaluate the antibiotic susceptibility of this anaerobic species. Fifty isolates of P. melaninogenica cultured from sputum of 50 PWCF have been included. Antibiotic susceptibility testing was performed using the agar diffusion method. All isolates were susceptible to the following antibiotics: amoxicillin/clavulanic acid, piperacillin/tazobactam, imipenem and metronidazole. A total of 96% of the isolates (48/50) were resistant to amoxicillin (indicating beta-lactamase production), 34% to clindamycin (17/50) and 24% to moxifloxacin (12/50). Moreover, 10% (5/50) were multidrug-resistant. A significant and positive correlation was found between clindamycin resistance and chronic azithromycin administration. This preliminary study on a predominant species of the lung “anaerobiome” shows high percentages of resistance, potentially exacerbated by the initiation of long-term antibiotic therapy in PWCF. The anaerobic resistome characterization, focusing on species rather than genera, is needed in the future to better prevent the emergence of resistance within lung microbiota.

Subgingival microbiome of deep and shallow periodontal sites in patients with rheumatoid arthritis: a pilot study

Background Subgingival microbiome in disease-associated subgingival sites is known to be dysbiotic and significantly altered. In patients with rheumatoid arthritis (RA), the extent of dysbiosis in disease- and health-associated subgingival sites is not clear. Methods 8 RA and 10 non-RA subjects were recruited for this pilot study. All subjects received full oral examination and underwent collection of subgingival plaque samples from both shallow (periodontal health-associated, probing depth ≤ 3mm) and deep subgingival sites (periodontal disease-associated, probing depth ≥ 4 mm). RA subjects also had rheumatological evaluation. Plaque community profiles were analyzed using 16 S rRNA sequencing. Results The phylogenetic diversity of microbial communities in both RA and non-RA controls was significantly higher in deep subgingival sites compared to shallow sites (p = 0.022), and the overall subgingival microbiome clustered primarily according to probing depth (i.e. shallow versus deep sites), and not separated by RA status. While a large number of differentially abundant taxa and gene functions was observed between deep and shallow sites as expected in non-RA controls, we found very few differentially abundant taxa and gene functions between deep and shallow sites in RA subjects. In addition, compared to non-RA controls, the UniFrac distances between deep and shallow sites in RA subjects were smaller, suggesting increased similarity between deep and shallow subgingival microbiome in RA. Streptococcus parasanguinis and Actinomyces meyeri were overabundant in RA subjects, while Gemella morbillorum, Kingella denitrificans, Prevotella melaninogenica and Leptotrichia spp. were more abundant in non-RA subjects. Conclusions The aggregate subgingival microbiome was not significantly different between individuals with and without rheumatoid arthritis. Although the differences in the overall subgingival microbiome was driven primarily by probing depth, in contrast to the substantial microbiome differences typically seen between deep and shallow sites in non-RA patients, the microbiome of deep and shallow sites in RA patients were more similar to each other. These results suggest that factors associated with RA may modulate the ecology of subgingival microbiome and its relationship to periodontal disease, the basis of which remains unknown but warrants further investigation.

Metagenome-genome-wide association studies reveal human genetic impact on the oral microbiome

The oral microbiota contains billions of microbial cells, which could contribute to diseases in a number of body sites. Challenged by eating, drinking and dental hygiene on a daily basis, the oral microbiota is regarded as highly dynamic. Here, we report significant human genomic associations with the oral metagenome from more than 1,915 individuals, for both the tongue dorsum and saliva. Five genetic loci, APPL2, SLC2A9 and MGST1 associated with tongue dorsum, LOC102723769-OR11H1-POTEH and MTRNR2L1-LOC105371703-MIR4522 associated with salivary microbial features, reached study-wide significance (p < 3.16 * 10-11). Further analyses confirmed 6 genome-wide significant loci shared between tongue dorsum and saliva. For example, the dental caries pathogen Prevotella melaninogenica associated with MARK2-RCOR2; the periodontitis bacteria Treponema associated with CCL26-CCL24 and Porphyromonas associated with CSMD1 at both niches. Human genetics account for at least 10% of oral microbiome differences between individuals. Machine learning models showed that polygenetic risk score dominated over oral microbiome in predicting predisposing risk of dental diseases such as dental calculus and gingival bleeding. These findings indicate that human genetic differences are one explanation for a stable or recurrent oral microbiome in each individual.

Inter-site and interpersonal diversity of salivary and tongue microbiomes, and the effect of oral care tablets

Background: Oral microbiota has been linked to both health and diseases. Specifically, tongue-coating microbiota has been implicated in aspiration pneumonia and halitosis. Approaches altering one's oral microbiota have the potential to improve oral health and prevent diseases. Methods: Here, we designed a study that allows simultaneous monitoring of the salivary and tongue microbiomes during an intervention on the oral microbiota. We applied this study design to evaluate the effect of single-day use of oral care tablets on the oral microbiome of 10 healthy individuals. Tablets with or without actinidin, a protease that reduces biofilm formation in vitro, were tested. Results: Alpha diversity of the tongue microbiome was significantly lower than that of the salivary microbiome, using both the number of observed amplicon sequence variants (254 ± 53 in saliva and 175 ± 37 in tongue; P = 8.9e-7, Kruskal–Wallis test) and Shannon index (6.0 ± 0.4 in saliva and 5.4 ± 0.3 in tongue; P = 2.0e-7, Kruskal–Wallis test). Fusobacterium periodonticum, Saccharibacteria sp. 352, Streptococcus oralis subsp. dentisani, Prevotella melaninogenica, Granulicatella adiacens, Campylobacter concisus, and Haemophilus parainfluenzae were the core operational taxonomic units (OTUs) common to both sites. The salivary and tongue microbiomes of one individual tended to be more similar to one another than to those of other individuals. The tablets did not affect the alpha or beta diversity of the oral microbiome, nor the abundance of specific bacterial species. Conclusions: While the salivary and tongue microbiomes differed significantly in terms of bacterial composition, they showed inter- rather than intra-individual diversity. A one-day usage of oral care tablets did not alter the salivary or tongue microbiomes of healthy adults. Whether the use of oral tablets for a longer period on healthy people or people with greater tongue coating accumulation shifts their oral microbiome needs to be investigated.

Salivary microbiome with gastroesophageal reflux disease and treatment

The effect of oral microbial composition on periodontal health and on systemic health has been, and is being established. The oral microbiome, in turn, can be altered by local and systemic diseases and conditions. Gastroesophageal reflux disease (GERD), has been associated with increased acidity in the oral cavity resulting in dental erosion, and controversially a reduced risk of periodontal disease. We hypothesized that presence of GERD was linked to a modified microbial profile in untreated GERD patients and that the use of proton pump inhibitor (PPI) drugs: potent disruptors of gut microbiome, in GERD patients might result in a salivary microbiome that is further distinct. Untreated GERD patients showed multiple differences in salivary microbiome as compared to healthy controls. Taxa found at lower levels related to the presence of GERD not treated by PPI included: Prevotella melaninogenica, Prevotella pallens, Leptotrichia, and Solobacterium moorei and thirteen others. In contrast, GERD patients chronically using PPI showed minimal differences in salivary taxa compared to healthy controls not using PPI.

Bactéria cromogênica associada a manchas negras

Objetivos: Este trabalho tem como objetivo avaliar a associação das bactérias Prevotella Melaninogênica e Actinomyces ao biofilme escuro em dentes permanentes. Relato de Caso: Um paciente que apresentou manchas enegrecidas ao longo do contorno cervical dos dentes foi convidado a participar do presente estudo. Após seu esclarecimento e aceitação, preenchendo o TCLE, foi coletado com o auxílio de uma cureta estéril, uma parte deste biofilme que foi inserido em um eppendorff contendo soro fisiológico. O material coletado foi agitado em um Vórtex por 15 segundos e em seguida, inoculadas em 100 microlitros (µl) da solução em um meio de ágar sangue suplementado com Canamicina 5%, seguido de cultivo por 72 horas. A partir do crescimento bacteriano, as mesmas foram isoladas em ágar macconkey, chocolate e sangue em anaerobiose. Em seguida foram realizados testes de bioquimismo, utilizando-se os meios MIO (motilidade, indol e ornitina), citrato, TSI (triplo açúcar com ferro) e LIA (ágar lisina com ferro). Conclusão: Através das características clínicas associadas ao bioquimismo empregado nos testes, podemos concluir que a bactéria que está associada as manchas negras presentes no paciente é do gênero prevotella spp.

Evaluation of the Microbiological Profile of Alveolar Residual Screws and Cleft-Adjacent Teeth in Individuals With Complete Unilateral Fissures

Objective: To evaluate the microbiota profile of residual alveolar slits and teeth adjacent to the cleft in fissured individuals. Designs: This study used a cross-sectional design. Participants: Twenty individuals, aged 14 to 24 years, who had a residual fissure in the maxillary alveolar ridge region were selected. Main outcome measures: Three sites per individual were selected for microbiological collection (the site of the residual cleft and the 2 nearest teeth). The samples were analyzed using the Checkerboard DNA–DNA hybridization technique for 73 species of bacteria. Results: All the species analyzed were found in the 2 niches (slits and teeth). The bacterial species present in the largest number in the residual cracks were Prevotella melaninogenica, Prevotella nigrescens, and Streptococcus mitis. With regard to the bacterial profiles in the mesial and distal faces, the most prevalent species were P nigrescens, Veillonella parvula, and Fusobacterium nucleatum sp vicentii. The analysis of all the collected samples demonstrated very similar profiles for the mesial and distal faces, with these 2 sites even presenting the same species in greater frequencies. Higher counts of 20 bacterial species (Wilcoxon test) were observed in the dental niche, in relation to the fissure, particularly, P nigrescens, V parvula, F nucleatum sp vicentii, and Neisseria mucosa. Conclusion: Some species were significantly more prevalent in the residual alveolar fissures and in adjacent teeth. The comparison between the profiles of the 2 niches demonstrated large differences in the most frequent species in the teeth, and no qualitative differences with regard to specific pathogens.