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急性牙髓感染患者不同口腔环境中的微生物生态学及预测代谢途径。

Microbial ecology and predicted metabolic pathways in various oral environments from patients with acute endodontic infections.

机构信息

Programa de Pós-graduação em Odontologia, Faculdade de Odontologia, Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil.

Curso de Odontologia, Universidade Regional Integrada do Alto Uruguai e das Missões, Erechim, Brazil.

出版信息

Int Endod J. 2020 Dec;53(12):1603-1617. doi: 10.1111/iej.13389. Epub 2020 Sep 22.

Abstract

AIM

To assess in a cross-sectional clinical study the effect of antibiotics on the diversity, structure and metabolic pathways of bacterial communities in various oral environments in patients with acute primary infections.

METHODOLOGY

Samples of saliva (SA), supragingival biofilm (SB) and from the pulp cavity (PC) were collected from teeth with acute primary infections and then grouped according to previous use of antibiotics (NoAtb = no antibiotics [n = 6]; Atb = antibiotics [n = 6]). DNA sequencing was conducted using MiSeq (Illumina, San Diego, CA, USA). The V1-V3 hyper-variable region of the 16S rRNA gene was amplified. A custom Mothur pipeline was used for 16S rRNA processing. Subsequent analyses of the sequence dataset were performed in R (using vegan, phyloseq and ggplot2 packages) or QIIME.

RESULTS

Twelve patients aged from 22 to 56 years were recruited. Participants in the Atb group had taken the beta-lactamics amoxicillin (5/6) or cephalexin (1/6) for 2-3 days. A total of 332 bacterial taxa (OTUs) were identified, belonging to 120 genera, 60 families and nine phyla. Firmicutes (41%) and Bacteroidetes (38%) were the most abundant phyla in all samples. Taxa clustered significantly by oral site (PCoA analysis; P < 0.05, ANOSIM). Use of antibiotics had little effect on this clustering. However, SA, SB and PC had different degrees of richness, diversity and evenness. The greatest diversity was observed in SB samples and the least diversity was observed in PC samples. Metabolic prediction identified 163 pathways and previous use of antibiotics had a major effect on the estimated functional clustering in SA and PC samples.

CONCLUSION

The ecological niche had a strong influence on the bacterial content of samples from various oral sites. Previous exposure to antibiotics may exert an effect on the phylogenetic composition of SA. Metabolic pathways appear to be modulated by antimicrobial agents in SA and PC samples. The dynamics of host/microbial interactions in the apical region and the functional ecology of the infected pulp cavity should be revisited.

摘要

目的

在一项横断面临床研究中评估抗生素对急性原发性感染患者不同口腔环境中细菌群落多样性、结构和代谢途径的影响。

方法

采集急性原发性感染牙齿的唾液(SA)、龈上生物膜(SB)和牙髓腔(PC)样本,然后根据抗生素使用情况进行分组(NoAtb = 无抗生素[ n = 6];Atb = 抗生素[ n = 6])。使用 MiSeq(Illumina,圣地亚哥,CA,美国)进行 DNA 测序。扩增 16S rRNA 基因的 V1-V3 高变区。使用定制的 mothur 管道进行 16S rRNA 处理。随后在 R(使用 vegan、phyloseq 和 ggplot2 包)或 QIIME 中对序列数据集进行分析。

结果

共招募了 12 名年龄在 22 至 56 岁之间的患者。Atb 组的参与者服用了β-内酰胺类的阿莫西林(5/6)或头孢氨苄(1/6),持续 2-3 天。共鉴定出 332 个细菌分类群(OTUs),属于 120 个属、60 个科和 9 个门。所有样本中最丰富的菌群是厚壁菌门(41%)和拟杆菌门(38%)。OTU 聚类分析表明口腔部位具有显著差异(PCoA 分析;P<0.05,ANOSIM)。抗生素的使用对这种聚类的影响很小。然而,SA、SB 和 PC 具有不同程度的丰富度、多样性和均匀度。SB 样本的多样性最大,PC 样本的多样性最小。代谢预测确定了 163 条途径,抗生素的使用对 SA 和 PC 样本中估计的功能聚类有很大影响。

结论

生态位对来自不同口腔部位样本的细菌含量有很强的影响。先前接触抗生素可能会对 SA 的系统发育组成产生影响。代谢途径似乎在 SA 和 PC 样本中被抗菌药物调节。根尖区宿主/微生物相互作用的动态和感染牙髓腔的功能生态学应重新审视。

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