Bernardi Sara, Continenza Maria Adelaide, Al-Ahmad Ali, Karygianni Lamprini, Follo Marie, Filippi Andreas, Macchiarelli Guido
Department of Life, Health and Environmental Sciences, University of L'Aquila, Via Vetoio, Coppito 2, 67100, L'Aquila, Italy.
Department of Operative Dentistry and Periodontology, Center of Dental Medicine, Oral and Maxillofacial Surgery, Medical Center, University of Freiburg, Hugstetter Straße, 55, 79106, Freiburg, German.
New Microbiol. 2019 Apr;42(2):108-113. Epub 2019 Apr 29.
The present study involved a qualitative and quantitative evaluation of tongue dorsum biofilms sampled from halitosis patients and healthy volunteers. The aim of the study was to quantify the distribution of Streptococcus spp. and Fusobacterium nucleatum within the oral halitosis biofilm in order to highlight the role of these bacterial members in halitosis. Tongue plaque samples from four halitosis-diagnosed patients and four healthy volunteers were analyzed and compared. The visualization and quantification of the tongue dorsum biofilm was performed combining fluorescence in situ hybridization (FISH) and confocal laser scanning microscopy (CLSM). Eubacteria, Streptococcus spp. and Fusobacterium nucleatum were stained using specific fluorescent probes. For a comparison of the two tested biofilm groups the Wilcoxon rank-sum test was used. Morphological analysis by CLSM illustrated the distribution of the species which were tracked. Streptococcus spp. appeared to be enclosed within the samples and always associated to F. nucleatum. Furthermore, compared to the control group the biofilm within the halitosis group contained significantly higher proportions of F. nucleatum and Streptococcus spp., as revealed by the FISH and CLSM-analysis. The total microbial load and relative proportions of F. nucleatum and Streptococcus spp. can be considered as causative factors of halitosis and thus, as potential treatment targets.
本研究对从口臭患者和健康志愿者采集的舌背生物膜进行了定性和定量评估。该研究的目的是量化口腔口臭生物膜中链球菌属和具核梭杆菌的分布，以突出这些细菌成分在口臭中的作用。对四名经口臭诊断的患者和四名健康志愿者的舌菌斑样本进行了分析和比较。结合荧光原位杂交（FISH）和共聚焦激光扫描显微镜（CLSM）对舌背生物膜进行可视化和定量分析。使用特异性荧光探针对真细菌、链球菌属和具核梭杆菌进行染色。为比较两个测试的生物膜组，使用了Wilcoxon秩和检验。CLSM的形态学分析说明了所追踪物种的分布。链球菌属似乎被包裹在样本中，并且总是与具核梭杆菌相关联。此外，FISH和CLSM分析显示，与对照组相比，口臭组内的生物膜中具核梭杆菌和链球菌属的比例显著更高。具核梭杆菌和链球菌属的总微生物负荷及相对比例可被视为口臭的致病因素，因此，可作为潜在的治疗靶点。
