Plaque Bacterial Microbiome Diversity in Children Younger than 30 Months with or without Caries Prior to Eruption of Second Primary Molars

Abstract

ObjectiveOur primary objective is to phylogenetically characterize the supragingival plaque bacterial microbiome of children prior to eruption of second primary molars by pyrosequencing method for studying etiology of early childhood caries.MethodsSupragingival plaque samples were collected from 10 caries children and 9 caries-free children. Plaque DNA was extracted, used to generate DNA amplicons of the V1–V3 hypervariable region of the bacterial 16S rRNA gene, and subjected to 454-pyrosequencing.ResultsOn average, over 22,000 sequences per sample were generated. High bacterial diversity was noted in the plaque of children with caries [170 operational taxonomical units (OTU) at 3% divergence] and caries-free children (201 OTU at 3% divergence) with no significant difference. A total of 8 phyla, 15 classes, 21 orders, 30 families, 41 genera and 99 species were represented. In addition, five predominant phyla (Firmicute, Fusobacteria, Proteobacteria, Bacteroidetes and Actinobacteria) and seven genera (Leptotrichia, Streptococcus, Actinomyces, Prevotella, Porphyromonas, Neisseria, and Veillonella) constituted a majority of contents of the total microbiota, independent of the presence or absence of caries. Principal Component Analysis (PCA) presented that caries-related genera included Streptococcus and Veillonella; while Leptotrichia, Selenomonas, Fusobacterium, Capnocytophaga and Porphyromonas were more related to the caries-free samples. Neisseria and Prevotella presented approximately in between. In both groups, the degree of shared organism lineages (as defined by species-level OTUs) among individual supragingival plaque microbiomes was minimal.ConclusionOur study represented for the first time using pyrosequencing to elucidate and monitor supragingival plaque bacterial diversity at such young age with second primary molar unerrupted. Distinctions were revealed between caries and caries-free microbiomes in terms of microbial community structure. We observed differences in abundance for several microbial groups between the caries and caries-free host populations, which were consistent with the ecological plaque hypothesis. Our approach and findings could be extended to correlating microbiomic changes after occlusion establishment and caries treatment.

Leptotrichia species in human infections II

“…clone, PCR, HOMIM[44]M/F 55 (±53.8–56.3) Leptotrichia sp. clonesHealthy and diseases (caries + periodontitis)Saliva, mouth A. xylosoxidan , M. micronuciformis , F. periodontium , V. atypica , PCR, HOMIM 124M/F < 30 Mo ±19.1 Leptotrichia spp., L. hongkongensis clonesCariesSupragingival plaque, mouth Porphyromonas , Corynebacterium, Capnocytophaga , Streptococcus spp., S. mutans clones, S. sobrinus clones, Veillonella, Neisseria , Rothia , TM7 genus incertae sedis , Actinomyces, Prevotella spp., P. histicola clones, Eikenella, Kingella , Fusobacterium, Gemella , Campylobacter, Granulicatella spp., G. adiacens clones, Abiotrophia, Selenomonas , Acinetobacter, Lactobacillus , Anaeroglobus, Ottowia , Schlegelella , HTS, PCA[45]M/F < 30 Mo ±19.0 Leptotrichia spp.CFSupragingival plaque, mouth Streptococcus spp., Capnocytophaga, Corynebacterium , TM7 genus incertae sedis , Porphyromonas, Granulicatella , Fusobacterium, Treponema , Gemella, Selenomonas , Veillonella, Dechloromonas , Actinomyces, Campylobacter , Abiotrophia , Ottowia , Eikenella, Johnsonella , Neisseria, Prevotella , Kingella, Rothia , HTS, PCA 126M/F/T 39 ± 10 Leptotrichia spp.HIV seropositiveSaliva Lactobacillus spp., Aggregatibacter, Lachnospiraceae , Rothia, Eubacterium , Tannerella, Haemophilus , Neisseria, Gemella , Granulicatella, Shuttleworthia , Streptococcus, S. mutans , Fusobacterium, Solobacterium , Campylobacter, Veillonella , Dialister , Synergistetes, Filifactor, Parvimonas , Achromobacter , Megasphaera, Selenomonas, Prevotella , Candida , culture, DGGE, HOMIM, PCA[46]M/F 43 ± 13 Leptotrichia spp.HIV seronegativeSaliva Capnocytophaga , Lachnospiraceae, Peptostreptococcaceae , Granulicatella, Veillonella , Synergistetes, Lactobacillus spp., Porphyromonas, Lactobacillus , Campylobacter, Streptococcus , S. mutans , Parvimonas, Kingella , Atopobium, Selenomonas , Aggregatibacter, Fusobacterium , Haemophilus, Megasphaera , Prevotella, Solobacterium , Gemella, Achromobacter , Eubacterium, Rothia , Slackia, Filifactor , Dialister, Neisseria , Candida , culture, DGGE, HOMIM, PCA 128M/F ≥ 18–21.5 ± 1.9 Leptotrichia spp., L. hongkongensis Healthy, supragingival plaqueMouth Corynebacterium , Capnocytophaga, Streptococcus , Cardiobacterium , Hae...…”

“…Since previous reviews [2,3], Leptotrichia species have been reported in >124 cases [4,7,16,18–69,87–93], whereby 30 cases involved L. buccalis [4,8,15,21,34,52,56,70–79,87], 24 cases L. wadei [4,20,24,34,37,42,48,56,67,80,81,90,93], 16 cases L. trevisanii [4,5,9,10,13,14,17,37,87], 14 cases L. hofstadii [34,40,49,56,81–84,93], 10 cases L. goodfellowii [4,11,12,21,56,74,85,87], eight cases L. hongkongensis [4,6,18,45,47,56], and five L. shahii [34,56,86]. L. trevisanii and L. wadei bacteremia are extremely rare; clinicians should consider these species in cases involving immunocompromised patients with oral lesions [4,5,13,17,87].…”

“…L. hongkongensis has been isolated from the blood of metastatic breast carcinoma (MBC) patients [6], the blood and amniotic fluid of a patient with a wound and respiratory difficulties [4], plaque from dental caries [45,47,56], saliva from pancreatic cancer patients and black pigmented stain caries patients (Table 2) [18,63]. …”

“…They were also recovered from oral plaque of guinea pigs [21] and feces of piglets [54], dental plaque from healthy individuals, plaque and saliva from patients with various types of caries, gingivitis, chronic periodontitis, and peri-implantitis [23,25–27,34,35,37,38,44,45,47,49,52,59,60,62,66–69,91,94], decayed tooth surfaces and discordant caries from intact enamel surfaces [53]. Leptotrichia species were also isolated from bronchoalveolar lavage fluid, and patients with leukocytosis, hypoxemia, and dyspnea [24].…”

Mutans Streptococci and Dental Caries: A New Statistical Modeling Approach

“…tween 7 and 36 months, and all studies linked MS detection to the time period coinciding with eruption of the primary teeth [Carlsson et al, 1975;Alaluusua and Renkonen, 1983;Fujiwara et al, 1991;Xu et al, 2014].…”