Most bacteria in natural and clinical settings grow as surface-attached biofilms, which are bacterial communities that have self-assembled into an encased matrix of extracellular polymeric substances (EPS). To form these bacterial biofilm communities and infect host cells, an intercellular signaling process described as quorum sensing (QS) is very important. For the opportunistic pathogen, Pseudomonas aeruginosa, QS regulates the expression of many genes important to biofilm initiation, EPS production, and virulence. While much has been learned about select factors that regulate biofilm formation in vitro and in animal models, the specifics by which multispecies groups of cells form biofilms are not yet clear. Correlated mass spectrometric and Raman imaging will be used to develop a multiplex analysis method for studying host-associated microbial communities. These methods allow the determination of individual bacterial species and their microbial products within a mixed bacterial community growing in situ on surfaces. A long-term goal is the design of detection and diagnostic strategies informed by an understanding of bacterial interactions and signature biomolecule production. Work toward this goal will begin by conducting experiments with the opportunistic pathogen Pseudomonas aeruginosa, specifically developing methods to distinguish P. aeruginosa from other bacterial species, distinguish separate P. aeruginosa strains from each other, and determine the spatial distribution of four types of P. aeruginosa molecules associated with virulence: acyl- homoserine lactones (AHL), quinolones, such as pseudomonas quinolone signal (PQS), rhamnolipids, and exopolymeric substances (EPS). Successfully implementing this research program will result in bacterial mapping methods with improved sensitivity and resolution, substantially enhancing our ability to identify specific species of bacteria in mixed samples, and to identify and map specific chemical products produced by these bacteria that are critical to host colonization, infection, and virulence.

Public Health Relevance

Most infections are the result of surface-attached biofilm communities of bacteria that colonize host surfaces and the bacterium Pseudomonas aeruginosa is an opportunistic pathogen that readily forms biofilms on many surfaces. We propose to use P. aeruginosa as a model species for developing correlated mass spectrometric and Raman imaging to perform multiplex analysis, enabling determination of individual bacterial species and many microbial products produced by these bacteria within a mixed bacterial community growing in situ on surfaces. We will use our new methodology to distinguish separate P. aeruginosa strains from each other, and determine the spatial distribution of acyl-homoserine lactone (AHL) and quinolone signaling molecules, biosurfactant rhamnolipids, and exopolymeric substances (EPS) produced by P. aeruginosa, each of which contributes to biofilm formation affecting virulence of P. aeruginosa.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI113219-05
Application #
9474586
Study Section
Enabling Bioanalytical and Imaging Technologies Study Section (EBIT)
Program Officer
Brown, Liliana L
Project Start
2014-06-01
Project End
2019-05-31
Budget Start
2018-06-01
Budget End
2019-05-31
Support Year
5
Fiscal Year
2018
Total Cost
Indirect Cost
Name
University of Notre Dame
Department
Engineering (All Types)
Type
Biomed Engr/Col Engr/Engr Sta
DUNS #
824910376
City
Notre Dame
State
IN
Country
United States
Zip Code
46556
Mattingly, Anne E; Weaver, Abigail A; Dimkovikj, Aleksandar et al. (2018) Assessing Travel Conditions: Environmental and Host Influences On Bacterial Surface Motility. J Bacteriol :
Dunham, Sage J B; Ellis, Joseph F; Baig, Nameera F et al. (2018) Quantitative SIMS Imaging of Agar-Based Microbial Communities. Anal Chem 90:5654-5663
Tomoshige, Shusuke; Dik, David A; Akabane-Nakata, Masaaki et al. (2018) Total Syntheses of Bulgecins A, B, and C and Their Bactericidal Potentiation of the ?-Lactam Antibiotics. ACS Infect Dis 4:860-867
Hu, Jiayun; Fu, Kaiyu; Bohn, Paul W (2018) Whole-Cell Pseudomonas aeruginosa Localized Surface Plasmon Resonance Aptasensor. Anal Chem 90:2326-2332
Morales-Soto, Nydia; Dunham, Sage J B; Baig, Nameera F et al. (2018) Spatially dependent alkyl quinolone signaling responses to antibiotics in Pseudomonas aeruginosa swarms. J Biol Chem 293:9544-9552
Si, Tong; Li, Bin; Comi, Troy J et al. (2017) Profiling of Microbial Colonies for High-Throughput Engineering of Multistep Enzymatic Reactions via Optically Guided Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry. J Am Chem Soc 139:12466-12473
Dunham, Sage J B; Ellis, Joseph F; Li, Bin et al. (2017) Mass Spectrometry Imaging of Complex Microbial Communities. Acc Chem Res 50:96-104
Hu, Jiayun; Bohn, Paul W (2017) Optical Biosensing of Bacteria and Bacterial Communities. J Anal Test 1:
Polisetti, Sneha; Baig, Nameera F; Morales-Soto, Nydia et al. (2017) Spatial Mapping of Pyocyanin in Pseudomonas Aeruginosa Bacterial Communities Using Surface Enhanced Raman Scattering. Appl Spectrosc 71:215-223
Si, Tong; Li, Bin; Zhang, Ke et al. (2016) Characterization of Bacillus subtilis Colony Biofilms via Mass Spectrometry and Fluorescence Imaging. J Proteome Res 15:1955-62

Showing the most recent 10 out of 16 publications