Co-localized or randomly distributed? Pair cross correlation of in vivo grown subgingival biofilm bacteria quantified by digital image analysis
Autor: | Annette Moter, Birgit Riep, Lawrence E. Wolinsky, Ulf B. Göbel, Anton Friedmann, Claudia Schillinger, Judith Kikhney, Holger Daims, Renate Lux, Annett Petrich |
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Přispěvatelé: | Semple, Malcolm Gracie |
Jazyk: | angličtina |
Rok vydání: | 2012 |
Předmět: |
Male
Bacterial Diseases Image Processing Colony Count Colony Count Microbial lcsh:Medicine Prevotella intermedia Image analysis Microbial ecology Microbial Computer-Assisted Oral Diseases Image Processing Computer-Assisted Pathology Bacteroides Tannerella forsythia Species interactions lcsh:Science 0303 health sciences Multidisciplinary biology medicine.diagnostic_test Perio-Endo Infectious Diseases Medical Microbiology Medicine Female Infection Porphyromonas gingivalis Research Article Clinical Pathology General Science & Technology Oral Medicine Microbiology Fluorescent in situ hybridization 03 medical and health sciences Forsythia Diagnostic Medicine In vivo medicine Humans Periodontal Pocket Dental/Oral and Craniofacial Disease Biology Periodontal Diseases 030304 developmental biology Fusobacterium nucleatum 030306 microbiology lcsh:R Biofilm Bacteriology biology.organism_classification Clinical Microbiology stomatognathic diseases Biofilms Dentistry Fusobacterium Infections Bacterial pathogens lcsh:Q Bacterial Biofilms Software Fluorescence in situ hybridization |
Zdroj: | PLoS ONE, Vol 7, Iss 5, p e37583 (2012) PLoS ONE PloS one, vol 7, iss 5 |
ISSN: | 1932-6203 |
Popis: | The polymicrobial nature of periodontal diseases is reflected by the diversity of phylotypes detected in subgingival plaque and the finding that consortia of suspected pathogens rather than single species are associated with disease development. A number of these microorganisms have been demonstrated in vitro to interact and enhance biofilm integration, survival or even pathogenic features. To examine the in vivo relevance of these proposed interactions, we extended the spatial arrangement analysis tool of the software daime (digital image analysis in microbial ecology). This modification enabled the quantitative analysis of microbial co-localization in images of subgingival biofilm species, where the biomass was confined to fractions of the whole-image area, a situation common for medical samples. Selected representatives of the disease-associated red and orange complexes that were previously suggested to interact with each other in vitro (Tannerella forsythia with Fusobacterium nucleatum and Porphyromonas gingivalis with Prevotella intermedia) were chosen for analysis and labeled with specific fluorescent probes via fluorescence in situ hybridization. Pair cross-correlation analysis of in vivo grown biofilms revealed tight clustering of F. nucleatum/periodonticum and T. forsythia at short distances (up to 6 µm) with a pronounced peak at 1.5 µm. While these results confirmed previous in vitro observations for F. nucleatum and T. forsythia, random spatial distribution was detected between P. gingivalis and P. intermedia in the in vivo samples. In conclusion, we successfully employed spatial arrangement analysis on the single cell level in clinically relevant medical samples and demonstrated the utility of this approach for the in vivo validation of in vitro observations by analyzing statistically relevant numbers of different patients. More importantly, the culture-independent nature of this approach enables similar quantitative analyses for "as-yet-uncultured" phylotypes which cannot be characterized in vitro. |
Databáze: | OpenAIRE |
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