Study of oral microbiota and diet as casual factors of Black Stains in the pediatric patient

The aim of the following study is to identify and quantify microorganisms in the saliva of black stain patients using species-specific probes for Actinomyces israelii, Actinomyces naeslundii, Streptococcus mutans and Lactobacillus spp, as well as a universal probe for bacterial 16S rDNA, to obtain the figure of the total bacterial counts (for test and control group). This study represents a first phase of the research line on extrinsic tooth pigmentation on developmental age patients. Materials and methods. The selected sample is composed of 20 subjects aged between 2 and 15 years and in good general and oral health among the patients included in a dental examination trial at the Department of Pediatric Dentistry of the University Hospital of Palermo, after obteining informed consent from parents. Subjects with a positive anamnesis for systemic diseases or with enamel defects were excluded from the study. Exclusion criteria were also a diet including regular use of tea, coffee, iron supplements, or antibiotic therapies in the 3 months prior to enrollment and the use of mouthwashes/toothpastes containing antiseptics. Parents/tutors were asked to complete a questionnaire on their occupational status and level of education, as well as their eating habits, the frequency of brushing and the patient's dental records. A sample of 20 patients without BS was used as control group. From all subjects participating to the study a sample of 5ml of unstimulated saliva was recovered from the right and left buccal area through a sterile syringe, after 2 minutes waiting for the accumulation of an adequate amount of saliva. Statistical analysis was carried out using descriptive and inferential statistics (mean and standard deviation; chi-square test, t test for non-paired data). StatView 5.0.1 statistical software (SAS Institute, Cary, NC, USA) was used for this purpose, and the tests were considered significant at p values less than or equal to 0.05. The study protocol will be presented for approval to the Palermo 1 ethics committee.