Abstract
ObjectiveThe current manuscript aims to determine the prevalence, duration and bacterial diversity of bacteraemia following dental extractions using conventional culture-dependent methods and 16S rDNA pyrosequencing.MethodsThe study group included 8 patients undergoing dental extractions under general anaesthesia. Peripheral venous blood samples were collected at baseline, 30 seconds and 15 minutes after the dental extractions. Blood samples were analysed for bacteraemia applying conventional microbiological cultures under aerobic and anaerobic conditions as well as pyrosequencing using universal bacterial primers that target the 16S ribosomal DNA gene.ResultsTransient bacteremia was detected by culture-based methods in one sample at baseline time, in eight samples at 30 seconds, and in six samples at 15 minutes after surgical procedure; whereas bacteraemia was detected only in five blood samples at 30 seconds after dental extraction by using pyrosequencing. By applying conventional microbiological methods, a single microbial species was detected in six patients, and Streptococcus viridans was the most frequently cultured identified bacterium. By using pyrosequencing approaches however, the estimated blood microbial diversity after dental extractions was 13.4±1.7 bacterial families and 22.8±1.1 genera per sample.ConclusionThe application of 16S rDNA pyrosequencing underestimated the prevalence and duration of bacteraemia following dental extractions, presumably due to not reaching the minimum DNA required for PCR amplification. However, this molecular technique, unlike conventional culture-dependent methods, revealed an extraordinarily high bacterial diversity of post-extraction bacteraemia. We propose that microorganisms recovered by culture may be only the tip of an iceberg of a really diverse microbiota whose viability and potential pathogenicity should be further studied.
Highlights
Bacteraemia is defined as the presence of bacteria in blood
A recent review of the literature revealed a prevalence of transient bacteraemia after dental extractions (BDE) that varies between 30% and 76% in children and between 58% and 100% in adults [6]
By applying 16S rDNA polymerase chain reaction (PCR) amplification (Figure 1) followed by pyrosequencing, the common bacterial diversity at family level could be identified in subgingival samples
Summary
Bacteraemia is defined as the presence of bacteria in blood. A feature that is unique to the oral bacterial biofilm, the subgingival plaque is its close proximity to a highly vascularised milieu. After reviewing published data of oral-derived bacteraemia significant differences were detected between studies in relation to the microbiological procedures applied, and to the transport and culture media, the atmosphere and incubation times used, and the characteristics of the isolates phenotypic identification process [6,13]. All these factors could affect bacterial isolation and identification ( of fastidious oral bacteria), and it has been stated that ‘‘oral bacteria recovered from blood by culture are probably only part of those present’’ [5]. The aim of the present study was to determine the prevalence, duration and bacterial diversity of bacteraemia following dental extractions using the conventional culture-based microbiological techniques and the 16S rDNA pyrosequencing approach, which is the first time that is applied to this kind of samples
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