Searchable abstracts of presentations at key conferences in endocrinology
Endocrine Abstracts (2016) 41 GP79 | DOI: 10.1530/endoabs.41.GP79

ECE2016 Guided Posters Diabetes (10 abstracts)

Metagenomic analysis of saliva and biofilm microbiome of type 1 diabetics under continuous subcutaneous insulin infusion

Eunice Carrilho 1, , Ana Coelho 2 , Anabela Paula 1 , Francisco Caramelo 1 , Mário Silva 2 , Francisco Carrilho 3 , Miguel Melo 1 , Luísa Barros 3 , Carla Batista 3 , Filomena Botelho 4 & Marques Ferreira 1,


1Faculty of Medicine, University of Coimbra, Coimbra, Portugal; 2Faculty of Dental Medicine, University of Porto, Porto, Portugal; 3Department of Endocrinology, Diabetes and Metabolism, Coimbra Hospital and University Centre, Coimbra, Portugal; 4Institute of Biomedical and Imaging and Life Sciences (IBILI), Coimbra, Portugal.


Introduction: The microvascular complications of diabetes and consequent autonomic neuropathy may cause changes in saliva’s secretion and composition. Hyperglycemic levels can also contribute to various oral pathological changes due to alteration of fungi or bacterial flora and dental biofilm surface, leading to the onset of candidiasis, periodontitis and dental caries. The aim of this study was to determine total bacterial load of saliva and dental biofilm of diabetic patients under continuous subcutaneous insulin infusion and that of non-diabetics.

Methods: Twenty patients with type 1 diabetes under continuous subcutaneous insulin infusion were included and 20 non-diabetic subjects were randomized as age-sex-matched controls. Unstimulated whole saliva and dental biofilm were collected from all patients under fasting conditions (at least 2 h). DNA was extracted from samples adding Bacteria Lysis Buffer (Roche) plus Proteinase K. The mixtures were incubated during 1 h at 65°C, followed by an in house extraction protocol based in DNA precipitation and isolation with cold etanol. Total bacterial load was quantified by Real-time PCR (qPCR) using primers targeting conserved bacterial 16S rDNA sequence in the LightCycler 2.0 instrument (Roche) by SYBR green detection.

Results: Diabetic patients had a significantly higher saliva and biofilm total bacterial load than non-diabetic subjects. No significant correlation was found between total bacterial load and glycated hemoglobin A1C.

Conclusion: This study suggests that type 1 diabetic patients have a higher saliva and biofilm total bacterial load than non-diabetic individuals, regardless of metabolic control. The knowledge of the diabetic oral microbiome may pave the way to the redefinition of clinical protocols and to the creation of new public health policies related to this population.

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