Variability of the fimA gene in Porphyromonas gingivalis isolated from periodontitis and non-periodontitis patients

Objective: The goal of this study was to determine the genetic variability of the fimA gene in Porphyromonas gingivalis isolates from Spanish patients. Study Design: Pooled subgingival samples were taken, processed and cultured in non-selective blood agar medium. Pure cultures of one to six isolates per patient were obtained and PCR and PCR-RFLP were used for fimbrillin gene (fimA) type determination of the extracted genomic (DNA). Results: Two hundred and twenty four Porphyromonas gingivalis isolates from 65 patients were analyzed consisting of 15 non-periodontitis patients (66 isolates) and 50 with periodontitis (158 isolates). Genotype II was the most prevalent (50.9%), while the other types of fimbriae did not exceed fifteen percent of prevalence. Isolates with types II and IV of fimbriae were significantly more prevalent in periodontitis patients than isolates with genotype I. Co-infection was observed in 17.65% of the patients analyzed. Conclusion: The results suggest that in this population Porphyromonas gingivalis with type II of fimbriae are significantly more predominant in periodontitis patients than genotype I

Detection and expression analysis of tet (B) in Streptococcus oralis

Tetracycline resistance can be achieved through tet genes, which code for efflux pumps, ribosomal protection proteins and inactivation enzymes. Some of these genes have only been described in either Gram-positive or Gram-negative bacteria. This is the case of tet (B), which codes for an efflux pump and, so far, had only been found in Gram-negative bacteria. In this study, tet (B) was detected in two clinical Streptococcus oralis strains isolated from the gingival sulci of two subjects. In both cases, the gene was completely sequenced, yielding 100% shared identity and coverage with other previously published sequences of tet (B). Moreover, we studied the expression of tet (B) using RT-qPCR in the isolates grown with and without tetracycline, detecting constitutive expression in only one of the isolates, with no signs of expression in the other one. This is the first time that the presence and expression of the tet (B) gene has been confirmed in Gram-positive bacteria, which highlights the potential of the genus Streptococcus to become a reservoir and a disseminator of antibiotic resistance genes in an environment so prone to horizontal gene transfer as is the oral biofilm

Tetracycline and multidrug resistance in the oral microbiota : differences between healthy subjects and patients with periodontitis in Spain

Introduction : Antibiotic resistance is widely found even among bacterial populations not having been exposed to selective pressure by antibiotics, such as tetracycline. In this study we analyzed the tetracycline-resistant subgingival microbiota of healthy subjects and of patients with periodontitis, comparing the prevalence of tet genes and their multidrug resistance profiles. Methods : Samples from 259 volunteers were analyzed, obtaining 813 tetracycline-resistant isolates. The prevalence of 12 antibiotic resistance genes was assessed, and multidrug profiles were built. Each isolate was identified by 16S rRNA sequencing. Differences in qualitative data and quantitative data were evaluated using the chi-square test and the Mann-Whitney-U test, respectively. Results : tet (M) was the most frequently detected tet gene (52.03%). We observed significant differences between the prevalence of tet (M), tet (W), tet (O), tet (32) and tet (L) in both populations studied. Multidrug resistance was largely observed, with resistance to kanamycin being the most detected (83.64%). There were significant differences between the populations in the prevalence of kanamycin, chloramphenicol, and cefotaxime resistance. Resistant isolates showed significantly different prevalence between the two studied groups. Conclusion : The high prevalence of multidrug resistance and tetracycline resistance genes found in the subgingival microbiota, highlights the importance of performing wider and more in-depth analysis of antibiotic resistance in the oral microbiota

Development and viability of biofilms grown on experimental abutments mimicking dental implants : an in vivo model

To determine whether an experimental abutment mimicking the macro- and microstructure of a dental implant is a suitable method for recovering biofilm, and to describe the features of biofilms formed around such abutments on healthy implants. Experimental abutments were used in 15 patients without peri-implant diseases. After 14 days? absence of dental hygiene in this area, the abutments were retrieved and analyzed through confocal laser scanning microscopy and scanning electron microscopy. The biofilm formation on the surface of the first 5 abutments was determined by a fluorescence-staining method using SYTO9 nucleic acid stain. In order to study the biofilm?s coverage and vitality, 10 additional abutments were assessed using live and dead bacterial viability. Descriptive and bivariate analyses of the data were performed. A global plaque coverage of the abutments was observed in all cases. The submucosal area of the abutment was mostly covered with biofilm (over 21%). Moreover, significant differences between supra- and subgingival locations were detected. This in vivo experimental model allows detailed observation of the extensive plaque growth found on exposed experimental abutments mimicking dental implants when hygiene measures are absent. The biofilm coverage is significantly higher in the supragingival zone than in the subgingival portion

Acyl homoserine lactone-mediated quorum sensing in the oral cavity: a paradigm revisited

Acyl homoserine lactones (AHLs), the quorum sensing (QS) signals produced by Gram-negative bacteria, are currently considered to play a minor role in the development of oral biofilm since their production by oral pathogens has not been ascertained thus far. However, we report the presence of AHLs in different oral samples and their production by the oral pathogen Porphyromonas gingivalis. The importance of AHLs is further supported by a very high prevalence of AHL-degradation capability, up to 60%, among bacteria isolated from dental plaque and saliva samples. Furthermore, the wide-spectrum AHL-lactonase Aii20J significantly inhibited oral biofilm formation in different in vitro biofilm models and caused important changes in bacterial composition. Besides, the inhibitory effect of Aii20J on a mixed biofilm of 6 oral pathogens was verified using confocal microscopy. Much more research is needed in order to be able to associate specific AHLs with oral pathologies and to individuate the key actors in AHL-mediated QS processes in dental plaque formation. However, these results indicate a higher relevance of the AHLs in the oral cavity than generally accepted thus far and suggest the potential use of inhibitory strategies against these signals for the prevention and treatment of oral diseasesThis work was supported by the grant “Axudas do Programa de Consolidación e Estructuración de Unidades de Investigación Competitivas (GPC)” from the Consellería de Cultura, Educación e Ordenación Universitaria, Xunta de Galicia (ED431B2017/53) and by the project INTERREG-POCTEP-0227-CODIGOMAIS-1-E. A.M. was supported by a predoctoral fellowship from the Consellería de Cultura, Educación e Ordenación Universitaria, Xunta de Galicia (ED481A-2015/311)S

Erythritol-enriched powder and oral biofilm regrowth on dental implants: an in vitro study.

BACKGROUND: Peri-implant mucositis and peri-implantitis are the main biological complications associated with dental implants. Since most authors agree that bacteria play a major etiological role, the main aims of this study were to determine if a formulation of erythritol and chlorhexidine applied with an air polishing system inhibits biofilm regrowth over dental implants and to compare the decontamination capacity of this therapy with that of mechanical removal by saline and gauze. MATERIAL AND METHODS: A multispecies biofilm (P. gingivalis, A. actinomycetemcomitans, F. nucleatum, A. naeslundii, V. parvula and S. oralis) was grown for 14 days on 52 dental implants in an artificial mouth. These implants were divided into three groups according to the applied treatment: 14 negative control (CON), 19 erythritol-chlorhexidine (ERY) and 19 gauze with saline (GAU) samples. Twelve dental implants from the ERY and GAU groups and 8 implants from the CON group were re-incubated for 7 additional days after treatment. The bacterial count was performed by quantitative polymerase chain reaction (qPCR) using propidium monoazide (PMA). A descriptive and bivariate analysis of the data was performed. RESULTS: The erythritol and chlorhexidine formulation significantly inhibited biofilm regrowth in comparison with the mechanical treatment (GAU), since a significant decrease in all the species was observed in the ERY group (except for Aggregatibacter actinomycetemcomitans). The antibiofilm and antibacterial capacity of the two active treatment groups (ERY and GAU) was similar for a 14 days multispecies in vitro biofilm, except for the lower count of A. naeslundii in the GAU group. CONCLUSIONS: The use of erythritol powder with chlorhexidine applied with an air polishing system reduces biofilm regrowth over dental implants when compared with mechanical removal by saline and gauze. This effect might be beneficial for patients included in peri-implant maintenance programs

Validation of ATP bioluminescence as a tool to assess antimicrobial effects of mouthrinses in an in vitro subgingival-biofilm model

Objectives. The aim of this investigation was to evaluate whether the adenosine triphosphate (ATP) bioluminescence method is an appropriate tool to assess the efficacy of antiseptic mouthrinses in terms of quantitative reductions of total viable microbial counts in mixed biofilm populations in vitro. Study Design. Three mouthrinses, containing respectively, chlorhexidine and cetylpyridinium chloride (CHX/CPC), essential oils (EO) and amine fluoride/stannous fluoride (AFSF), as well as Phosphate Buffered Saline (PBS) used as control, were tested in an in vitro static biofilm model by ATP bioluminescence and compared to culture method. Biofilms were grown on saliva-coated hydroxyapatite disks for 72 hours and then exposed for 1 minute to the mouthrinse or control by immersion. The antibacterial effect of the rinses was tested by analysis of variance. The reliability of the ATP bioluminescence method was assessed by calculating the Pearson correlation coefficients when compared to the viable cell counts obtained by culture. Results. Using ATP bioluminescence, the antimicrobial activity of the tested mouthrinses was demonstrated when compared to the PBS control. The ATP bioluminescence values were significantly correlated (0.769, p<0.001) to the viable cell counts. CHX/CPC and AFSF showed similar antimicrobial activity, although AFSF had a less homogeneous effect, being both more effective than the EO rinse. Conclusion. ATP bioluminescence viability testing may be considered a useful tool to assess the in vitro efficacy of antibacterial compounds. In the proposed model, CHX/CPC and AFSF containing mouthrinses demonstrated superior antimicrobial activity, as compared to EO rinses, in a multispecies biofilm model

Microbial profile of placentas from Tanzanian mothers with adverse pregnancy outcomes and periodontitis

Aim: To investigate microbial profiles in placentas from a population of East African mothers with and without adverse pregnancy outcomes and with regard to their periodontal status. Material and Methods: Thirty-six placentas from pregnant women from Tanzania were classified into three groups according to both pregnancy outcome and the mother's periodontal health. The microbial composition in each group was then compared using 16S rRNA metagenomics. Additionally, placenta specimens were analyzed histologically for chorioamnionitis by a single pathologist blinded to the clinical data. Results: The greatest differences were observed in the group of mothers with periodontitis. The microbial load was low in all three groups of mothers. Periodontitis had a notable influence on the structure of the placental microbiota. Three phyla and 44 genera were associated with periodontitis, whereas only the Tenericutes phylum was associated with the adverse pregnancy variable. Streptococcaceae and Mycoplasmataceae families were associated with both periodontitis and adverse pregnancy outcomes. Finally, although the differences for chorioamnionitis were not significant, this intra-amniotic infection was more frequent in the placentas from mothers with periodontitis

Maturational stages: comparison of growth and physical capacity indicators in adolescents

Introduction: The identification of physical capacity is an important marker related to healthy behavior during childhood and adolescence, in which some factors appear to contribute to motor performance such as maturation and hormonal levels. Objective: To compare growth indicators, physical capacity and hormonal markers according to gender and maturational stage in adolescents. Methods: Eighty-nine adolescents of both genders aged 10-13 years participated in the study. Sexual maturation was evaluated using the Tanner’s self-evaluation method. Physical capacity (explosive strength of upper and lower limbs, upper limb velocity and agility) and hormonal markers (testosterone and estradiol) were evaluated through the chemiluminescence method. Results: In the comparison by gender, girls had higher weight (p = 0.023), height (p = 0.018) and fat percentage values (p = 0.001), while boys presented better motor performance for the explosive strength of upper limbs (p = 0.005) and lower limbs (p = 0.011), agility (0.018) and upper limb velocity (p = 0.014). Regarding maturational stage, boys did not present differences in any variable analyzed; (Stage V versus I), height (stage III, IV and V versus I) and upper limb explosive strength (stage III and IV versus I). Conclusion: Growth, weight and height, as well as explosive strength of upper limbs were higher in girls at more advanced maturational stages and appear to be gender dependent

Utilização de diferentes métodos para a determinação da idade óssea em jovens

O estudo tem como objetivo correlacionar os resultados da idade óssea em meninos e meninas em diferentes métodos. Na metodologia para verificar a idade óssea, foram realizadas aferições de estatura, diâmetros de úmero e fêmur, perímetro corrigido de braço, dobra tricipital, idade cronológica para compor o modelo matemático, e raio-x de mão e punho para utilização do método de Grave e Brown (1976). No tratamento estatístico realizado no programa SPSS 20.0 utilizando estatística descritiva, correlação de Pearson e diferença significativa entre os métodos. As correlações obtiveram resultados positivos para meninos e meninas, respectivamente, com resultados entre raio x e idade cronológica (M=0,84/F=0,53), modelo matemático (M=0,85/F=0,56) e estatura (M=0,80/F=0,28), assim como entre o modelo matemático e estatura (M=0,95/ F=0,84) e idade cronológica (M=0,95/F=0,76). Concluímos, desta forma, existir correlação entre os métodos, não existindo diferença significativa entre os mesmos, destacando assim o modelo matemático por ser um método prático e de fácil aplicação para uso no meio esportivo