Microbiological techniques
FRANKLIN
The number of colony forming units (cfu) was determined by swabbing the cheek mucosa on the right and on the left side at tooth 36 level and tooth 46 level. A slight pressure was applied to the mucous membrane and the area of sampling was limited to 1 cm
Sterilizable stainless steel template for standardized sampling within an area of 1 cmAfter sampling, the swab was flushed in 0.9% NaCl and the supernatant plated on Columbia agar plates (BD Becton Dickinson Corporation, Franklin Lakes, NJ, USA). One part of the saliva sample was mixed with nine parts on a previously validated inactivator containing 3% Tween 80, 0.3% lecithin, 0.1% histidine and 0.1% sodium thiosulfate. The respective dilutions were plated onto Columbia blood agar plates following incubation for 48 h at 37 °C in a microaerophilic atmosphere. Afterwards, aerobic cultivated cfu (total bacteria count without distinction of colony morphology, species, etc.) were counted.