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1.
PLoS One ; 11(9): e0163001, 2016.
Article in English | MEDLINE | ID: mdl-27631785

ABSTRACT

Enterococci are considered as transient constituent components of the oral microbiome that may cause a variety of oral and systemic infections. As there is sparse data on the oral enterococcal prevalence, we evaluated the Enterococcus spp. and their virulence attributes including antimicrobial resistance in a healthy Brazilian cohort. A total of 240 individuals in different age groups were studied (children 4-11 yrs, adolescents 12-17 yrs, young adults 18-29 yrs, adults 30-59 yrs, elderly over 60 yrs). Oral rinses were collected and isolates were identified by API 20 Strep and confirmed by 16S rDNA sequencing. E. faecalis isolates, in particular, were evaluated for virulence attributes such as their biofilm formation potential, and susceptibility to antimicrobials and an antiseptic, chlorhexidine gluconate. A total of 40 individuals (16.6%) and 10% children, 4% adolescents, 14% young adults, 30% adults, and 25% elderly carried oral enterococci. The oral enterococcal burden in adolescents was significantly lower than in the adults (p = 0.000) and elderly (p = 0.004). The proportion of carriers was higher among females (p = 0.001). E. faecalis was the most frequent isolate in all the age groups (p = 0.000), followed by E. durans and E. faecium. Whilst all the clinical isolates were able to form biofilms, only a proportion of them were able to produce lipase (92%), hemolysin (38%), and gelatinase (39%). Of all the isolates 53.8% were resistant to tetracycline, 12.3% to amoxicillin, 16.0% to ampicillin, 20.8% to chloramphenicol and 43.4% to erythromycin. None of the isolates were resistant to vancomycin. Our data suggest that in this Brazilian cohort the oral cavity may act as a significant reservoir of rather virulent and antibiotic resistant enterococci, with an increasing degree of carriage in the adults and elderly. Hence clinicians should be cognizant of this silent reservoir of virulent enterococci that may pose a particular threat of nosocomial infection.


Subject(s)
Enterococcus/isolation & purification , Mouth/microbiology , Adolescent , Adult , Brazil , Child , Child, Preschool , Cohort Studies , Enterococcus/drug effects , Enterococcus/genetics , Enterococcus/pathogenicity , Genes, Bacterial , Humans , Microbial Sensitivity Tests , Middle Aged , RNA, Ribosomal, 16S/genetics , Virulence , Young Adult
2.
Environ Health Prev Med ; 20(5): 354-9, 2015 Sep.
Article in English | MEDLINE | ID: mdl-26108971

ABSTRACT

OBJECTIVES: The familial clustering observed in chronic kidney disease of uncertain etiology (CKDu) characterized by tubulointerstitial damages in the North Central Region of Sri Lanka strongly suggests the involvement of genetic factors in its pathogenesis. The objective of the present study is to use whole-exome sequencing to identify the genetic variants associated with CKDu. METHODS: Whole-exome sequencing of eight CKDu cases and eight controls was performed, followed by direct sequencing of candidate loci in 301 CKDu cases and 276 controls. RESULTS: Association study revealed rs34970857 (c.658G > A/p.V220M) located in the KCNA10 gene encoding a voltage-gated K channel as the most promising SNP with the highest odds ratio of 1.74. Four rare variants were identified in gene encoding Laminin beta2 (LAMB2) which is known to cause congenital nephrotic syndrome. Three out of four variants in LAMB2 were novel variants found exclusively in cases. CONCLUSION: Genetic investigations provide strong evidence on the presence of genetic susceptibility for CKDu. Possibility of presence of several rare variants associated with CKDu in this population is also suggested.


Subject(s)
Exome , Genetic Predisposition to Disease/genetics , Renal Insufficiency, Chronic/genetics , Case-Control Studies , Genetic Predisposition to Disease/epidemiology , Humans , Laminin/genetics , Laminin/metabolism , Male , Polymorphism, Single Nucleotide , Renal Insufficiency, Chronic/epidemiology , Risk Factors , Shaker Superfamily of Potassium Channels/genetics , Shaker Superfamily of Potassium Channels/metabolism , Sri Lanka/epidemiology
3.
J Investig Clin Dent ; 6(1): 32-9, 2015 Feb.
Article in English | MEDLINE | ID: mdl-25154801

ABSTRACT

AIM: No oral niche can be considered to be segregated from the subjacent milieu because of the complex community behavior and nature of the oral biofilms. The aim of this study was to address the paucity of information on how these species are clonally related to the subjacent gingival crevice bacteria. METHODS: We utilized a metagenomic approach of amplifying 16S rDNA from genomic DNA, cloning, sequencing and analysis using LIBSHUFF software to assess the genetic homogeneity of the bacterial species from two infected root canals and subjacent gingival crevices. RESULTS: The four niches studied yielded 186 clones representing 54 phylotypes. Clone library comparisons using LIBSHUFF software indicated that each niche was inhabited by a unique flora. Further, 42% of the clones were of hitherto unknown phylotypes indicating the extent of bacterial diversity, especially in infected root canals and subjacent gingival crevices. CONCLUSIONS: We believe data generated through this novel analytical tool shed new light on understanding oral microbial ecosystems.


Subject(s)
Bacteria/classification , Dental Plaque/microbiology , Dental Pulp Cavity/microbiology , Gingiva/microbiology , Microbial Consortia/physiology , Adult , Bacteria/genetics , Biodiversity , DNA, Bacterial/analysis , Dental Pulp Exposure/microbiology , Genome, Microbial/genetics , Humans , Incisor/injuries , Male , Metagenome/genetics , Periapical Diseases/microbiology , Phylogeny , Polymerase Chain Reaction , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Tooth Fractures/microbiology
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