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The aim of this study is to compare the efficacy of two methods for collecting saliva samples from infants under 2 years of age for cariogenic streptococci (CS) count. Two collection methods were applied in 11 infants. In Method (A), saliva samples were collected by swabbing the inner cheek mucosa and floor of the mouth in figure of eight motions with a sterile cotton swab until it was soaked. In method (B), saliva samples were collected by aspiration of 1 ml of saliva with a sterile plastic syringe on the floor of the mouth, after stimulation with glove. The samples were cultured in modified Gold’s broth (MSMG), and on trypticase, yeast extract, sucrose, cystine and bacitracin culture medium (TYSCB). In method (A), the swab with the sample was unloaded in situ on TYSCB and placed in PBS medium for transport. Then, 100 μl of the eluate was seeded in MSMG. In method (B) 100 μl were seeded in TYSCB and 100 μl in MSMG. Both culture media were incubated under capnophilic conditions for 48 hours at 37 °C. Colony forming units (CFU/ml) were counted by calibrated operators (kappa = 0.75). The presence of cariogenic streptococci (CS) (Streptococcus mutans-Streptococcus sobrinus) was determined by qPCR in the samples collected by both methods. The CFU/ml counts in MSMG differed significantly between methods (p = 0.021). In TYSCB, the recovery of CFU/ml was higher in method (A), without significant difference (p = 0.705). The molecular technique detected presence of CS, with no difference between collection methods. Collecting saliva samples by swabbing proved more effective in terms of recovery of microorganisms, and did not affect the detection of presence of CS by molecular techniques.

A strong association between periodontitis and higher susceptibility to pregnancy complications like preeclampsia has been reported although the mechanisms remain elusive. Trophoblast cells modulate the recruitment and functional shaping of maternal leukocytes at early stages to sustain an antiinflammatory microenvironment and fetal growth. Neutrophil activation with reactive oxygen species (ROS) release is associated with preeclampsia. Our aim was to study the effect of the gingival crevicular fluid (GCF) from pregnant women on trophoblast cell function and trophoblast-neutrophil interaction.Pregnant women at 16-20 weeks of gestation (n = 27) and non-pregnant women (n = 8) as the control group were studied for gingivoperiodontal clinical score evaluation and GCF collection. Total bacteria and common periodontal pathogens were analyzed in GCF samples. The effect of each GCF sample was tested on first trimester trophoblast-derived cells to assess cell migration, cytokine expression and glucose uptake. Also, the effect of GCF on human peripheral neutrophil chemoattraction by trophoblast cells and ROS formation was assessed.Gingival crevicular fluid from pregnant women reduced trophoblast cell migration, increased proinflammatory marker expression and glucose uptake. A significant correlation between gingivoperiodontal score and trophoblast dysfunction was observed. Upon conditioning of trophoblast cells with GCF, only the GCF from pregnant women stimulated neutrophil chemoattraction. Similarly, GCF from pregnant but not from non-pregnant controls stimulated ROS formation in neutrophils.Gingival crevicular fluid from pregnant women is deleterious for first trimester trophoblast cell function. These effects could lead to placental homeostasis disruption underlying a pathogenic mechanism of pregnancy complications associated to periodontal disease.