Accepted Manuscript Quantitation of biofilm and planktonic life forms of coexisting periodontal species Maribasappa Karched, Radhika Bhardwaj, Anandavalli Inbamani, Sirkka Asikainen PII:

S1075-9964(15)30014-7

DOI:

10.1016/j.anaerobe.2015.04.013

Reference:

YANAE 1438

To appear in:

Anaerobe

Received Date: 4 February 2015 Revised Date:

21 April 2015

Accepted Date: 24 April 2015

Please cite this article as: Karched M, Bhardwaj R, Inbamani A, Asikainen S, Quantitation of biofilm and planktonic life forms of coexisting periodontal species, Anaerobe (2015), doi: 10.1016/ j.anaerobe.2015.04.013. This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.

RI PT

ACCEPTED MANUSCRIPT

M AN U

SC

Quantitation of biofilm and planktonic life forms of coexisting periodontal species

Maribasappa Karched, Radhika Bhardwaj, Anandavalli Inbamani, Sirkka Asikainen* Oral Microbiology, General Facility Laboratory, Faculty of Dentistry, Kuwait University, Kuwait.

TE D

Running title: Multispecies biofilms of periodontal species

AC C

EP

*Corresponding author Professor Sirkka Asikainen Department of Bioclinical Sciences Faculty of Dentistry Kuwait University PO Box 24923 Safat 13110 Kuwait Tel: +965-24636807 Email: [email protected]

1

ACCEPTED MANUSCRIPT Abstract Background: Complexity of oral polymicrobial communities has prompted a need for developing in vitro models to study behavior of coexisting bacteria. Little knowledge is available of in vitro co-growth of several periodontitis-associated species without early colonizers of dental plaque.

RI PT

The aim was to determine temporal changes in the quantities of six periodontal species in an in vitro biofilm model in comparison with parallel planktonic cultures.

Material and methods: Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Prevotella intermedia, Parvimonas micra, Campylobacter rectus and

SC

Fusobacterium nucleatum were anaerobically grown as multispecies and monospecies biofilms and parallel planktonic cultures using cell culture plates and microfuge tubes,

M AN U

respectively. After incubating 2, 4, 6, 8 days, biofilms and planktonic cultures were harvested, DNA extracted and the target species quantified using qPCR with speciesspecific 16S rDNA primers. Biofilm growth as monocultures was visualized at day 2 and 8 with confocal microscopy and crystal violet staining.

Results: The six species were found throughout the test period in all culture conditions, except that P. gingivalis and F. nucleatum were not detected in multispecies planktonic

TE D

cultures at day 8. In multispecies biofilm, P. gingivalis qPCR counts (cells/ml) increased (P

Quantitation of biofilm and planktonic life forms of coexisting periodontal species.

Complexity of oral polymicrobial communities has prompted a need for developing in vitro models to study behavior of coexisting bacteria. Little knowl...
9MB Sizes 6 Downloads 15 Views

Recommend Documents