Cryptic Oral Microbiota: What Is Its Role as Obstructive Sleep Apnea-Related Periodontal Pathogens?

Periodontitis has been commonly linked to periodontopathogens categorized in Socransky's microbial complexes; however, there is a lack of knowledge regarding "other microorganisms" or "cryptic microorganisms", which are rarely thought of as significant oral pathogens and have been neither previously categorized nor connected to illnesses in the oral cavity. This study hypothesized that these cryptic microorganisms could contribute to the modulation of oral microbiota present in health or disease (periodontitis and/or obstructive sleep apnea (OSA) patients). For this purpose, the presence and correlation among these cultivable cryptic oral microorganisms were identified, and their possible role in both conditions was determined. Data from oral samples of individuals with or without periodontitis and with or without OSA were obtained from a previous study. Demographic data, clinical oral characteristics, and genera and species of cultivable cryptic oral microorganisms identified by MALDI-TOF were recorded. The data from 75 participants were analyzed to determine the relative frequencies of cultivable cryptic microorganisms' genera and species, and microbial clusters and correlations tests were performed. According to periodontal condition, dental-biofilm-induced gingivitis in reduced periodontium and stage III periodontitis were found to have the highest diversity of cryptic microorganism species. Based on the experimental condition, these findings showed that there are genera related to disease conditions and others related to healthy conditions, with species that could be related to different chronic diseases being highlighted as periodontitis and OSA comorbidities. The cryptic microorganisms within the oral microbiota of patients with periodontitis and OSA are present as potential pathogens, promoting the development of dysbiotic microbiota and the occurrence of chronic diseases, which have been previously proposed to be common risk factors for periodontitis and OSA. Understanding the function of possible pathogens in the oral microbiota will require more research.

Patients with obstructive sleep apnea can favor the predisposing factors of periodontitis by the presence of P. melaninogenica and C. albicans, increasing the severity of the periodontal disease.

Objective: The aim of this study was to analyze the cultivable oral microbiota of patients with obstructive sleep apnea (OSA) and its association with the periodontal condition. Methods: The epidemiology profile of patients and their clinical oral characteristics were determined. The microbiota was collected from saliva, subgingival plaque, and gingival sulcus of 93 patients classified into four groups according to the periodontal and clinical diagnosis: Group 1 (n = 25), healthy patients; Group 2 (n = 17), patients with periodontitis and without OSA; Group 3 (n = 19), patients with OSA and without periodontitis; and Group 4 (n = 32), patients with periodontitis and OSA. Microbiological samples were cultured, classified, characterized macroscopically and microscopically, and identified by MALDI-TOF-MS. The distribution of complexes and categories of microorganisms and correlations were established for inter- and intra-group of patients and statistically evaluated using the Spearman r test (p-value <0.5) and a multidimensional grouping analysis. Result: There was no evidence between the severity of OSA and periodontitis (p = 0.2813). However, there is a relationship between the stage of periodontitis and OSA (p = 0.0157), with stage III periodontitis being the one with the highest presence in patients with severe OSA (prevalence of 75%; p = 0.0157), with more cases in men. The greatest distribution of the complexes and categories was found in oral samples of patients with periodontitis and OSA (Group 4 P-OSA); even Candida spp. were more prevalent in these patients. Periodontitis and OSA are associated with comorbidities and oral conditions, and the microorganisms of the orange and red complexes participate in this association. The formation of the dysbiotic biofilm was mainly related to the presence of these complexes in association with Candida spp. Conclusion: Periodontopathogenic bacteria of the orange complex, such as Prevotella melaninogenica, and the yeast Candida albicans, altered the cultivable oral microbiota of patients with periodontitis and OSA in terms of diversity, possibly increasing the severity of periodontal disease. The link between yeasts and periodontopathogenic bacteria could help explain why people with severe OSA have such a high risk of stage III periodontitis. Antimicrobial approaches for treating periodontitis in individuals with OSA could be investigated in vitro using polymicrobial biofilms, according to our findings.

Fumigant toxicity and biochemical effects of selected essential oils toward the red flour beetle, Tribolium castaneum (Coleoptera: Tenebrionidae).

Tribolium castaneum Herbst (Coleoptera: Tenebrionidae) is an insect popularly known as the red flour beetle, it is widely distributed worldwide and can cause serious damage to stored grains. Chemical control is the most used method for managing this pest, however, some substances are toxic to mammals and the environment. Therefore, the development of new effective and safe insecticides is necessary. Essential oils (EOs) can be considered as a potential alternative in the development of pesticides due to their physicochemical properties and varied effects against insects. In the current study, was determined the fumigant toxicity and biochemical effects of selected essential oils against T. castaneum. The 23 selected EOs were characterized by GC-MS and their fumigant lethal concentrations were determined. An exploratory Cluster analysis was performed to find a relationship between fumigant toxicity and chemical composition. Finally, the inhibition of the catalytic activity of acetylcholinesterase (AChE), glutathione S-transferase (GST) and catalase (CAT) was evaluated using protein homogenates obtained from T. castaneum. The results indicated that EOs with the highest fumigant potential were those with greater diversity in their composition, while the least active EOs presented mainly monoterpenes. The most active EOs were those obtained from Foeniculum vulgare and Zanthoxylum monophyllum with LC50 values of 16.23 and 18.54 µL/L air respectively. Regarding the inhibition of the enzymatic activity of the 23 EOs evaluated at 500 µL/L, only two caused an inhibition greater that 50% on AChE, which corresponded to EOs from Piper nigrum and Rosmarinus officinalis. Likewise, EOs from C. sinensis, Piper aduncum and Zanthoxylum monophyllum were the only ones able to inhibiting GST activity by more than 50%. Respecting CAT inhibition, 7 EOs caused and inhibition greater than 50%, highlighting those from Lavandula angustifolia, C. sempervirens and Eucalyptus sp. These results show that the EOs evaluated in this study seems to be a promising bio-controller of T. castaneum since have high fumigant toxicity and exert different mechanisms of action.

Effects of Essential Oils from 24 Plant Species on Sitophilus zeamais Motsch (Coleoptera, Curculionidae).

Chemical control of the maize weevil (Sitophilus zeamais) has been ineffective and presents serious collateral damage. Among plant-derived insecticides, essential oils (EOs) are suitable candidates to control this stored products pest. In this work, the insecticidal activities of 45 natural EOs against S. zeamais adults were screened, and the most promising ones (24 EOs) were characterized by GC-MS. The repellent and toxic effects (contact and fumigant) of these 24 EOs were determined, and by a cluster analysis they were classified into two groups considering its fumigant activity and contact toxicity. For the EOs with the highest fumigant potential (14 oils) and their main active constituents (17 compounds), lethal concentrations were determined. The most active EOs were those obtained from L. stoechas and L. alba, with LC50 values of 303.4 and 254.1 µL/L air and characterized by a high content of monoterpenes. Regarding the major compounds, the oxygenated monoterpenes R-(+)-pulegone (LC50 = 0.580 mg/L air), S-(-)-pulegone (LC50 = 0.971 mg/L air) and R-(-)-carvone (LC50 = 1.423 mg/L air) were the most active, as few variations in their concentrations significantly increased insect mortality.