Microbiology Commons^™

A Biofilm Matrix-Associated Protease Inhibitor Protects Pseudomonas Aeruginosa From Proteolytic Attack, Boo Shan Tseng, Courtney Reichhardt, Gennifer E. Merrihew, Sophia A. Araujo-Hernandez, Joe J. Harrison, Michael J. Maccoss, Matthew R. Parsek

Life Sciences Faculty Research

Pseudomonas aeruginosa produces an extracellular biofilm matrix that consists of nucleic acids, exopolysaccharides, lipid vesicles, and proteins. In general, the protein component of the biofilm matrix is poorly defined and understudied relative to the other major matrix constituents. While matrix proteins have been suggested to provide many functions to the biofilm, only proteins that play a structural role have been characterized thus far. Here we identify proteins enriched in the matrix of P. aeruginosa biofilms. We then focused on a candidate matrix protein, the serine protease inhibitor ecotin (PA2755). This protein is able to inhibit neutrophil elastase, a bactericidal enzyme …

Nitric Oxide Inhibits Biofilm Formation By Vibrio Fischeri Via The Nitric Oxide Sensor Hnox, Cecilia Thompson

Dissertations

Nitric oxide (NO) is an important signal secreted by the squid, Euprymna scolopes, and is thought to limit symbiotic biofilm formation, the initial step in colonization by the bacterium, Vibrio fischeri. Previous work demonstrated that a mutant defective for NO-sensor HnoX (heme nitric oxide/oxygen binding) had a competitive advantage over wild-type V. fischeri, but the mechanism by which this occurred remains unknown. HnoX is encoded upstream of hahK, a recently identified positive regulator of biofilm formation. I thus hypothesized that HnoX inhibits colonization by controlling HahK-induced biofilm formation during the initiation of colonization.

I assessed the impact of an hnoX …

The 'Pseudomonas Aeruginosa' Psl Polysaccharide Is A Social But Noncheatable Trait In Biofilms, Yasuhiko Irie, Aled E. Roberts, Kasper N. Kragh, Vernita D. Gordon, Jaime B. Hutchison, Rosalind J. Allen, Gavin Melaugh, Thomas Bjarnsholt, Stuart A. West, Stephen P. Diggle

Biology Faculty Publications

Extracellular polysaccharides are compounds secreted by microorganisms into the surrounding environment, and they are important for surface attachment and maintaining structural integrity within biofilms. The social nature of many extracellular polysaccharides remains unclear, and it has been suggested that they could function as either cooperative public goods or as traits that provide a competitive advantage. Here, we empirically tested the cooperative nature of the PSL polysaccharide, which is crucial for the formation of biofilms in Pseudomonas aeruginosa. We show that (i) PSL is not metabolically costly to produce; (ii) PSL provides populationlevel benefits in biofilms, for both growth and antibiotic …

Are Cdi Systems Multicolored, Facultative, Helping Greenbeards?, Elizabeth S. Danka, Erin C. Garcia, Peggy A. Cotter

Microbiology, Immunology, and Molecular Genetics Faculty Publications

Competitive and cooperative interactions between organisms, including bacteria, can significantly impact the composition of a community and the fitness of its members, as well as the fitness of their hosts when communities are living on or within other organisms. Understanding the underlying mechanisms is critical to the development of strategies to control microbiological communities that impact animal and plant health and also for understanding the evolution of social behaviors, which has been challenging for evolutionary biologists. Contact-dependent growth inhibition (CDI) is a phenomenon defined by the delivery of a protein toxin to the cytoplasm of neighboring bacteria upon cell–cell contact, …

Burkholderia Cenocepacia J2315-Mediated Destruction Of Staphylococcus Aureus Nrs77 Biofilms., Rachel Thompson

College of Arts & Sciences Senior Theses

Cystic fibrosis (CF) is an inherited disorder that affects over 30,000 people in the US and more than 70,000 people worldwide. Recurring bacterial infections in CF patients result in tissue damage that dramatically lowers respiratory function and are ultimately fatal. The formation of bacterial biofilms in the mucus and on lung epithelial tissue allows pathogens to be protected from antibiotics and the host immune system, making treatment of infection difficult. The interactions between CF pathogens in co-culture biofilms are not well understood and were examined in this study. Staphylococcus aureus and Burkholderia cenocepacia, two common CF pathogens, were used …

Multiscale Modeling Of Bacterial Colonies: How Pili Mediate The Dynamics Of Single Cells And Cellular Aggregates, Wolfram Pönisch, Christoph A. Weber, Guido Juckeland, Nicolas Biais, Vasily Zaburdaev

Publications and Research

Neisseria gonorrhoeae is the causative agent of one of the most common sexually transmitted diseases, gonorrhea. Over the past two decades there has been an alarming increase of reported gonorrhea cases where the bacteria were resistant to the most commonly used antibiotics thus prompting for alternative antimicrobial treatment strategies. The crucial step in this and many other bacterial infections is the formation of microcolonies, agglomerates consisting of up to several thousands of cells. The attachment and motility of cells on solid substrates as well as the cell–cell interactions are primarily mediated by type IV pili, long polymeric filaments protruding from …

Novel Approaches To Mitigating Bacterial Biofilm Formation And Intercellular Communication, Stephen Kasper

Legacy Theses & Dissertations (2009 - 2024)

Long thought of as solitary single-cell organisms, it is now widely accepted that bacteria can act and cooperate as social organisms. Phenomena such as biofilm formation and quorum sensing (QS) are two intimately intertwined cooperative behaviors that significantly contribute to the pathogenesis of many bacteria. Biofilms are surface associated communities of bacteria encased in a secreted extracellular matrix, which provides several advantages over an individualized lifestyle, such as increased protection from antimicrobial agents as well as enhanced opportunity for the exchange of genetic material. Bacterial QS is a system of population-based communication through the production, sensing, and response to chemical …

The Pseudomonas Aeruginosa Efflux Pump Mexghi-Opmd Transports A Natural Phenazine That Controls Gene Expression And Biofilm Development, Hassan Sakhtah, Leslie Koyama, Yihan Zhang, Diana K. Morales, Blanche Fields, Alexa Price-Whelan, Deborah Hogan

Dartmouth Scholarship

Redox-cycling compounds, including endogenously produced phenazine antibiotics, induce expression of the efflux pump MexGHI-OpmD in the opportunistic pathogen Pseudomonas aeruginosa Previous studies of P. aeruginosa virulence, physiology, and biofilm development have focused on the blue phenazine pyocyanin and the yellow phenazine-1-carboxylic acid (PCA). In P. aeruginosa phenazine biosynthesis, conversion of PCA to pyocyanin is presumed to proceed through the intermediate 5-methylphenazine-1-carboxylate (5-Me-PCA), a reactive compound that has eluded detection in most laboratory samples. Here, we apply electrochemical methods to directly detect 5-Me-PCA and find that it is transported by MexGHI-OpmD in P. aeruginosa strain PA14 planktonic and biofilm cells. We …

Spatiotemporal Control Of Staphylococcus Aureus Biofilm Development, Derek E. Moormeier

Theses & Dissertations

Chronic biofilm-related infections caused by the human pathogen Staphylococcus aureus often lead to significant increases in morbidity and mortality in both hospital- and community-associated settings. Typically, S. aureus biofilm development occurs in three stages: 1) attachment, 2) tower maturation, and 3) dispersal. Here, utilizing BioFlux1000 time-lapse microscopy we have expanded upon these fundamental stages of biofilm development and also unveiled and characterized two additional stages (multiplication and exodus).

The attachment and multiplication stages were shown to be protease sensitive but independent of most cell surface-associated proteins. Following multiplication, an exodus of the biofilm population that followed the transition of the …

Identification Of Novel Cyclic Lipopeptides From A Positional Scanning Combinatorial Library With Enhanced Antibacterial And Antibiofilm Activities, Nina Bionda, Renee M. Fleeman, César De La Fuente-Núñez, Maria C. Rodriguez, Fany Reffuveille, Lindsey N. Shaw, Irena Pastar, Stephen C Davis, Robert E W Hancock, Predrag Cudic

Molecular Biosciences Faculty Publications

Treating bacterial infections can be difficult due to innate or acquired resistance mechanisms, and the formation of biofilms. Cyclic lipopeptides derived from fusaricidin/LI-F natural products represent particularly attractive candidates for the development of new antibacterial and antibiofilm agents, with the potential to meet the challenge of bacterial resistance to antibiotics. A positional-scanning combinatorial approach was used to identify the amino acid residues responsible for driving antibacterial activity, and increase the potency of these cyclic lipopeptides. Screening against the antibiotic resistant ESKAPE pathogens revealed the importance of hydrophobic as well as positively charged amino acid residues for activity of this class …

Investigating The Regulatory Mechanisms By Which Arabinose And Sypa Control Biofilm Formation In Vibrio Fischeri, Sheila Paris

Master's Theses

Biofilms are surface-associated microbial communities surrounded by an extracellular matrix. Through biofilm formation, many pathogens and symbionts are able to colonize, survive, and persist in their host. A model system used to study biofilm formation is the symbiotic bacterium Vibrio fischeri, which colonizes its host, the squid Euprymna scolopes. Complex signaling between the squid and the bacteria is essential for the proper regulation of biofilm formation as well as for persistent colonization.

The signal(s) that promote host-relevant biofilm formation are as-yet unknown, but recently it was discovered that the sugar, L-arabinose, serves as a unique signal to promote biofilm formation …

Land-Use And Algal Species Influence Bacterial Community Composition And Activity In Streams, Miguel Rojas

Master's Theses

Microbial biofilms can be found on virtually all submerged surfaces in aquatic habitats, including rivers and streams. Biofilms are complex, taxonomically diverse communities of microorganisms (including algae and bacteria) that colonize solid surfaces and support a wide range of metabolic activities. Biofilms are critical components of river and stream ecosystems because they are key drivers of nutrient cycling processes (including nitrogen cycling) and because they serve as an important food source for higher trophic levels. Biofilms in urban streams are subject to a number of anthropogenic stressors, but research on the ecology of urban biofilms and their response to stressors …

Functional Characterization Of Aggregatibacter Actinomycetemcomitans Qsebc : A Bacterial Adrengeric Receptor And Global Regulator Of Virulence., Whitney Weigel

Electronic Theses and Dissertations

In order for a pathogen to successfully colonize the host, it must be able to acquire essential nutrients and regulate gene expression to respond to environmental fluctuations. One mechanism bacteria have evolved in order to detect these fluctuations and respond is the two component signal transduction systems (TCS). Aggregatibacter actinomycetemcomitans, a dental commensal associated with localized aggressive periodontitis, contains a TCS designated QseBC that was previously shown to regulate expression of several iron acquisition genes, biofilm formation and virulence, but the activating signal was unknown []. QseBC is a conserved TCS in the Enterobacteriaceae and Pasteurellaceae, but the activating signal …

The Effects Of Electrostatic Spraying With Organic Acids In The Disintegration Of Biofilms Formed By E.Coli O157:H7 And Salmonella Typhimurium On Spinach And Cantaloupe, Ahmad Almasoud

Graduate Theses and Dissertations

Outbreaks from the consumption of fresh produce are a concern in the United States. The consumptions of fresh produce have increased recently which expose a large segment of society to such outbreaks. Spinach and cantaloupe are minimally heated or processed before consumption which makes them a possible source of foodborne illness. The objective of this research was to investigate the effect of organic acids alone and in combination to reduce attached Salmonella Typhimurium (S.T) and Escherichia coli O157:H7 (E.coli) on spinach and cantaloupe, and to disintegrate biofilm formed by these pathogens by electrostatically spraying with two organic acids. To quantify …

Candida Albicans Ethanol Stimulates Pseudomonas Aeruginosa Wspr-Controlled Biofilm Formation As Part Of A Cyclic Relationship Involving Phenazines, Annie I. Chen, Emily F. Dolben, Chinweike Okegbe, Colleen E. Harty, Yuriy Golub, Sandy Thao, Dae Gon Ha, Sven D. Willger, George A. O'Toole, Caroline S. Harwood, Lars E. P Dietrich, Deborah A. Hogan

Dartmouth Scholarship

In chronic infections, pathogens are often in the presence of other microbial species. For example, Pseudomonas aeruginosa is a common and detrimental lung pathogen in individuals with cystic fibrosis (CF) and co-infections with Candida albicans are common. Here, we show that P. aeruginosa biofilm formation and phenazine production were strongly influenced by ethanol produced by the fungus C. albicans. Ethanol stimulated phenotypes that are indicative of increased levels of cyclic- di-GMP (c-di-GMP), and levels of c-di-GMP were 2-fold higher in the presence of ethanol. Through a genetic screen, we found that the diguanylate cyclase WspR was required for ethanol …

Structural Features Of The Pseudomonas Fluorescens Biofilm Adhesin Lapa Required For Lapg-Dependent Cleavage, Biofilm Formation, And Cell Surface Localization, Chelsea D. Boyd, T. Jarrod Smith, Sofiane El-Kirat-Chatel, Peter D. Newell, Yves F. Dufrêne, George A. O'Toole

Dartmouth Scholarship

The localization of the LapA protein to the cell surface is a key step required by Pseudomonas fluorescens Pf0-1 to irreversibly attach to a surface and form a biofilm. LapA is a member of a diverse family of predicted bacterial adhesins, and although lacking a high degree of sequence similarity, family members do share common predicted domains. Here, using mutational analysis, we determine the significance of each domain feature of LapA in relation to its export and localization to the cell surface and function in biofilm formation. Our previous work showed that the N terminus of LapA is required for …

Investigating The Impact Of Novel Iron Oxide Nanoparticles On Legionella Pneumophila Biofilms And Trophic Interactions, Brennen Jenkins

All Theses

Microbial biofilms serve as the base of food webs and are important for nutrient cycling in aquatic ecosystems. Nanoparticles (NPs) that enter into these aquatic systems have the potential to settle and become trapped within biofilms. As NPs become further integrated into consumer products, understanding their fate and effects on aquatic ecosystems is of paramount importance. Previous studies from our lab show that gold NPs induce dispersal of Legionella pneumophila biofilms. NPs with platinum and iron oxide core chemistries also lead to similar dispersal events, however, silver core NPs do not seem to induce these events due to NP aggregation. …

Deletion Mutant Library For Investigation Of Functional Outputs Of Cyclic Diguanylate Metabolism In Pseudomonas Aeruginosa Pa14, Dae-Gon Ha, Megan E. Richman, George A. O'Toole

Dartmouth Scholarship

We constructed a library of in-frame deletion mutants targeting each gene in Pseudomonas aeruginosa PA14 predicted to participate in cyclic di-GMP (c-di-GMP) metabolism (biosynthesis or degradation) to provide a toolkit to assist investigators studying c-di-GMP-mediated regulation by this microbe. We present phenotypic assessments of each mutant, including biofilm formation, exopolysaccharide (EPS) production, swimming motility, swarming motility, and twitch motility, as a means to initially characterize these mutants and to demonstrate the potential utility of this library.

Control Of Candida Albicans Metabolism And Biofilm Formation By Pseudomonas Aeruginosa Phenazines, Diana K. Morales, Nora Grahl, Chinweike Okegbe, Lars E. P. Dietrich, Nicholas J. Jacobs, Deborah A. Hogan

Dartmouth Scholarship

Candidaalbicanshasdevelopmentalprogramsthatgoverntransitionsbetweenyeastandfilamentousmorphologies and between unattached and biofilm lifestyles. Here, we report that filamentation, intercellular adherence, and biofilm develop- ment were inhibited during interactions between Candida albicans and Pseudomonas aeruginosa through the action of P. aeruginosa-produced phenazines. While phenazines are toxic to C. albicans at millimolar concentrations, we found that lower concentrations of any of three different phenazines (pyocyanin, phenazine methosulfate, and phenazine-1-carboxylate) allowed growth but affected the development of C. albicans wrinkled colony biofilms and inhibited the fungal yeast-to-filament transition. Phenazines impaired C. albicans growth on nonfermentable carbon sources and led to increased production of fer- mentation products (ethanol, glycerol, and …

Characterization Of The Novel Response Regulator Sype: A Dual Regulator Of Biofilm Formation And Colonization In Vibrio Fischeri, Andrew Robert Morris

Dissertations

Biofilms, or surface-associated microbial communities encased in a matrix, represent a common mode of growth for many bacterial species. Importantly, biofilms often play significant roles in the association of bacteria with their animal hosts, including promoting attachment to and colonization of host tissues. The symbiosis between the bacterium Vibrio fischeri and its animal host Euprymna scolopes provides a natural model system for investigating the impact of biofilms on host infection. In particular, V. fischeri produces a specific biofilm to successfully initiate host colonization.

Biofilm formation by V. fischeri is a tightly regulated process involving a complex network of regulatory genes. …

Microfluidic-Assisted Atomic Force Microscopy For The Mechanical Characterization Of Soft Biological Materials, Aaron Peter Mosier

Legacy Theses & Dissertations (2009 - 2024)

Viable methods for bacterial biofilm remediation require a fundamental understanding of biofilm mechanical properties and their dependence on dynamic environmental conditions. Mechanical test data, quantifying elasticity or adhesion, may be used to perform physical modeling of biofilm behavior, thus enabling the development of novel remediation strategies. To achieve real-time, dynamic measurements of these properties, a novel analysis platform consisting of a microfluidic flowcell device has been designed and fabricated for in situ analysis using atomic force microscopy (AFM) and confocal laser scanning microscopy (CLSM). The flowcell consists of microfluidic channels for biofilm establishment that are then converted into an open …

The Evolution Of Host Specificity In The Vertebrate Gut Symbiont Lactobacillus Reuteri, Steven Frese

Department of Food Science and Technology: Dissertations, Theses, and Student Research

The vertebrate gut is home to one of the densest populations of life on Earth. This microbial community has a profound effect on host health, nutrition, development, behavior, and evolution. However, very little is known about how these microbes have evolved with their vertebrate hosts, how and whether they select hosts or how they remain associated with their hosts. Recent work identified Lactobacillus reuteri as an organism that is composed of host-specific sub-populations, each population associated with a different host animal. Representatives from each host-associated population were tested for their ability to colonize gnotobiotic mice, which only rodent strains could …

Form And Function Of Clostridium Thermocellum Biofilms, Alexandru Dumitrache, Gideon Wolfaardt, Grant Allen, Steven N. Liss, Lee R. Lynd

Dartmouth Scholarship

The importance of bacterial adherence has been acknowledged in microbial lignocellulose conversion studies; however, few reports have described the function and structure of biofilms supported by cellulosic substrates. We investigated the organization, dynamic formation, and carbon flow associated with biofilms of the obligately anaerobic cellulolytic bacterium Clostridium thermocellum 27405. Using noninvasive, in situ fluorescence imaging, we showed biofilms capable of near complete substrate conversion with a characteristic monolayered cell structure without an extracellular polymeric matrix typically seen in biofilms. Cell division at the interface and terminal endospores appeared throughout all stages of biofilm growth. Using continuous-flow reactors with a rate …

In Vitro Antimicrobial Effect Of A Cold Plasma Jet Against Enterococcus Faecalis Biofilms, Chunqi Jiang, Christoph Schaudinn, David E. Jaramillo, Paul Webster, J. William Costerton

Bioelectrics Publications

The hypothesis that a cold plasma jet has the antimicrobial effect against Enterococcus faecalis biofilms was tested in vitro. 27 hydroxyapatite discs were incubated with E. faecalis for six days to form a monoculture biofilm on the disc surface. The prepared substrata were divided into three groups: the negative control, the positive control (5.25% NaOCl solution), and the plasma treatment group. Resultant colony-forming unit counts were associated with observations of bacterial cell morphology changes using scanning electron microscopy (SEM). Treatment of E. faecalis biofilm with the plasma and 5.25% NaOCl for 5min resulted in 93.1% and 90.0% kill (P < 0.0001), respectively. SEM detected that nearly no intact bacteria were discernible for the plasmaexposed HA disc surfaces. The demonstrated bactericidal effect of the plasma with direct surface contact may be due to the enhanced oxidation by the locally produced reactive plasma species.

Toxin-Antitoxic Loci Vapbc-1 And Vapxd Contribute To Survival And Virulence In Nontypeable Haemophilus Influenzae, Dabin Ren, Anna N. Walker, Dayle A. Daines

Biological Sciences Faculty Publications

Background: Nontypeable Haemophilus influenzae (NTHi) is a significant human pathogen responsible for respiratory tract infections and the most common cause of recurrent otitis media. Type II toxin-antitoxin (TA) systems are genetic elements that code for a stable protein toxin and a labile antitoxin that are thought to be involved in metabolic regulation of bacteria by enabling a switch to a dormant state under stress conditions. The contribution to infection persistence of the NTHi TA loci vapBC-1 and vapXD was examined in this study.

Results: Deletions in vapBC-1, vapXD and vapBC-1 vapXD significantly decreased the survival of NTHi co-cultured with …

Effects Of Larval Exposure To Bacterial Species In Three Genera On Life History Traits And Oviposition Behavior In The Southern House Mosquito, Culex Quinquefasciatus, Jonathan Gravgaard

All Theses

The biofilm is the most common niche for microorganisms living in aquatic environments. Forming a biofilm provides the means to resist shear forces, adverse osmolarity, chemical agents, and other environmental stressors. Biofilms also provide a major food source for aquatic arthropods like mosquito larvae. Culex quinquefasciatus larvae are found in freshwater pools with high organic content, and have the ability to graze on bacterial biofilms. To study how C. quinquefasciatus mosquitoes interact with bacteria in their environment I developed an experimental system that allows mosquito larvae to freely graze on bacteria in either the planktonic or biofilm state. The efficacy …

Systematic Analysis Of Diguanylate Cyclases That Promote Biofilm Formation By Pseudomonas Fluorescens Pf0-1, Peter D. Newell, Shiro Yoshioka, Kelli L. Hvorecny, Russell D. Monds, George A. O'Toole

Dartmouth Scholarship

Cyclic di-GMP (c-di-GMP) is a broadly conserved, intracellular second-messenger molecule that regulates biofilm formation by many bacteria. The synthesis of c-di-GMP is catalyzed by diguanylate cyclases (DGCs) containing the GGDEF domain, while its degradation is achieved through the phosphodiesterase activities of EAL and HD-GYP domains. c-di-GMP controls biofilm formation by Pseudomonas fluorescens Pf0-1 by promoting the cell surface localization of a large adhesive protein, LapA. LapA localization is regulated posttranslationally by a c-di-GMP effector system consisting of LapD and LapG, which senses cytoplasmic c-di-GMP and modifies the LapA protein in the outer membrane. Despite the apparent requirement for c-di-GMP for …

Novel Model For Multispecies Biofilms That Uses Rigid Gas-Permeable Lenses, Rebecca Peyyala, Sreenatha S. Kirakodu, Jeffrey L. Ebersole, Karen F. Novak

Center for Oral Health Research Faculty Publications

Oral biofilms comprise complex multispecies consortia aided by specific inter- and intraspecies interactions occurring among commensals and pathogenic bacterial species. Oral biofilms are primary initiating factors of periodontal disease, although complex multifactorial biological influences, including host cell responses, contribute to the individual outcome of the disease. To provide a system to study initial stages of interaction between oral biofilms and the host cells that contribute to the disease process, we developed a novel in vitro model system to grow biofilms on rigid gas-permeable contact lenses (RGPLs), which enable oxygen to permeate through the lens material. Bacterial species belonging to early- …

Non-Identity-Mediated Crispr-Bacteriophage Interaction Mediated Via The Csy And Cas3 Proteins, Kyle C. Cady, George A. O'Toole

Dartmouth Scholarship

Studies of the Escherichia, Neisseria, Thermotoga, and Mycobacteria clustered regularly interspaced short palindromic repeat (CRISPR) subtypes have resulted in a model whereby CRISPRs function as a defense system against bacteriophage infection and conjugative plasmid transfer. In contrast, we previously showed that the Yersinia-subtype CRISPR region of Pseudomonas aeruginosa strain UCBPP-PA14 plays no detectable role in viral immunity but instead is required for bacteriophage DMS3-dependent inhibition of biofilm formation by P. aeruginosa. The goal of this study is to define the components of the Yersinia-subtype CRISPR region required to mediate this bacteriophage-host interaction. We show that the Yersinia-subtype-specific CRISPR-associated (Cas) proteins …

Aminoglycoside Resistance Of Pseudomonas Aeruginosa Biofilms Modulated By Extracellular Polysaccharide, Wajiha Khan, Steve P. Bernier, Sherry L. Kuchma, John H. Hammond, Fariha Hasan, George A. O'Toole

Dartmouth Scholarship

Pseudomonas aeruginosa is an opportunistic pathogen that produces sessile communities known as biofilms that are highly resistant to antibiotic treatment. Limited information is available on the exact role of various components of the matrix in biofilm-associated antibiotic resistance. Here we show that the presence of extracellular polysaccharide reduced the extent of biofilm-associated antibiotic resistance for one class of antibiotics. Minimal bactericidal concentration (MBC) for planktonic and biofilm cells of P. aeruginosa PA14 was measured using a 96 well microtiter plate assay. The MBC of biofilm-grown ΔpelA mutant, which does not produce the Pel polysaccharide, was 4-fold higher for tobramycin and …