A method for growing a biofilm under low shear at the air–liquid interface using the drip flow biofilm reactor

Goeres, Darla M.; Hamilton, Martin A.; Beck, Nicholas A.; Buckingham‐Meyer, Kelli; Hilyard, Jackie D; Loetterle, Linda R.; Lorenz, Lindsey A.; Walker, Diane K.; Stewart, Philip S.

doi:10.1038/nprot.2009.59

Cited by 193 publications

(158 citation statements)

References 14 publications

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“…Some organisms may respond differently to varying types of fluid and flow combinations, including the drip rate of the liquid, material of the substratum, exposure to gases (or lack thereof) and experiment length. Ceramic or metal substrates can be used when simulating the oral cavity, and catheter, endotracheal tube lumens, and other biomedically relevant polymer substrates, are all potential surfaces that can be used with drip flow reactors [35,56].…”

Section: Drip Flow Reactormentioning

confidence: 99%

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Biofilm Models of Polymicrobial Infection

2015

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Interactions between microbes are complex and play an important role in the pathogenesis of infections. These interactions can range from fierce competition for nutrients and niches to highly evolved cooperative mechanisms between different species that support their mutual growth. An increasing appreciation for these interactions, and desire to uncover the mechanisms that govern them, has resulted in a shift from monomicrobial to polymicrobial biofilm studies in different disease models. Here we provide an overview of biofilm models used to study select polymicrobial infections and highlight the impact that the interactions between microbes within these biofilms have on disease progression. Notable recent advances in the development of polymicrobial biofilm-associated infection models and challenges facing the study of polymicrobial biofilms are addressed. Polymicrobial interactions in biofilmsOver the past 2 decades there has been a revolutionary paradigm shift in the field of microbiology with the appreciation that bacteria present in most biological systems exist in biofilms, rather than in a free-living state. This understanding has dramatically changed the way we study bacteria in the laboratory and resulted in the development of many new experimental systems that replicate biofilm environments [1][2][3][4][5]. Studies utilizing these systems have demonstrated time and time again that bacteria behave very differently when in a biofilm than during planktonic growth. In many ways, we now have to relearn everything we thought we knew about bacterial behavior through the view of the biofilm lens.Similarly, the recent explosion of metagenomic studies has significantly increased our appreciation of the complexity of the microbial populations present in biofilms [6,7]. This is also true for infections, most of which are thought to be biofilm related and inherently polymicrobial, including various species of bacteria, fungi and viruses [8]. It is thought that microbes act in concert to establish biofilms, which in turn can increase tolerance to antimicrobials, exacerbate of the host's immune response and increase persistence at the infection site [7,[9][10][11].Genetic diversity of microbes within biofilm communities is thought to increase the fitness of the residing community, making them more equipped to survive environmental stresses. In large part, this is due to an expanded gene pool, which can be more easily shared within the confines of a biofilm community [12]. Community composition and interactions within the community can have huge influences on bacterial behavior. Thus, just as the behavior of planktonic versus biofilm-associated bacteria is dramatically different, so is that of bacteria in single species versus multispecies systems.Interactions between microbes are complex and highly dependent on context. They can range from fierce competition for nutrients and niches, manifested by antagonistic behavior, to highly evolved cooperative mechanisms between different species that support their mutual grow...

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Section: Drip Flow Reactormentioning

confidence: 99%

“…Biofilm models of polymicrobial infection review future science group www.futuremedicine.com endotracheal tube lumens, and other biomedically relevant polymer substrates, are all potential surfaces that can be used with drip flow reactors [35,56].…”

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confidence: 99%

Biofilm Models of Polymicrobial Infection

2015

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“…Biofilms were grown using a modified DFR (model 110; Biosurface Technologies Corporation) on glass microscope slides (25675 mm) as described by Goeres et al (2009). Each slide was washed with detergent, rinsed with distilled water, dried and placed in the DFR.…”

Section: Determination Of Optimal Antimicrobial Agent Concentrationmentioning

confidence: 99%

Evidence for persisters in Staphylococcus epidermidis RP62a planktonic cultures and biofilms

Shapiro

Nguyen

Chamberlain

2011

Journal of Medical Microbiology

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The pathogenesis of Staphylococcus epidermidis in foreign device-related infections is attributed primarily to its ability to form biofilms on a polymer surface. One mechanism proposed for the survival of organisms in a biofilm is the presence of persister cells. Persister cells survive antibiotic treatment without acquiring heritable antibiotic resistance. This study was conducted to determine if S. epidermidis RP62a growing in planktonic cultures and biofilms could survive as persister cells following treatment with levofloxacin and vancomycin. S. epidermidis RP62a produced a small percentage of persisters (levofloxacin, 3.09¾10 "7 %; vancomycin, 8.21¾10 "5 %) when grown to exponential phase, whereas biofilms contained 28 and 94 % persisters, following exposure to levofloxacin and vancomycin, respectively. The highest percentages of persisters were obtained during stationary phase in planktonic cultures and the lowest percentages of persisters were obtained during mid-exponential phase. An increase in persister number was not due to activation of quorum-sensing regulons. Confocal laser scanning microscopy images of biofilms exposed to levofloxacin demonstrated that the antibiotic was able to kill bacteria throughout the biofilm. Our results suggest that antibiotic tolerance in biofilms and in planktonic cultures of S. epidermidis RP62a is due in part to the presence of persister cells.

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“…To further analyze the LPS, CPS, and PGA mutants, we examined biofilm formation in the drip-flow reactor, a continuousflow model with an air-liquid interface and low shear force considered to be more representative of the lung environment (35). While the O-antigen mutant barely formed detectable thin biofilms, parental strain 4074Nal r , LPS core mutant CG3, and capsule mutant 33.2 formed similar amounts of thick, rough biofilms (Fig.…”

Section: Resultsmentioning

confidence: 99%

Surface Polysaccharide Mutants Reveal that Absence of O Antigen Reduces Biofilm Formation of Actinobacillus pleuropneumoniae

et al. 2016

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c Actinobacillus pleuropneumoniae is a Gram-negative bacterium belonging to the Pasteurellaceae family and the causative agent of porcine pleuropneumonia, a highly contagious lung disease causing important economic losses. Surface polysaccharides, including lipopolysaccharides (LPS) and capsular polysaccharides (CPS), are implicated in the adhesion and virulence of A. pleuropneumoniae, but their role in biofilm formation is still unclear. In this study, we investigated the requirement for these surface polysaccharides in biofilm formation by A. pleuropneumoniae serotype 1. Well-characterized mutants were used: an Oantigen LPS mutant, a truncated core LPS mutant with an intact O antigen, a capsule mutant, and a poly-N-acetylglucosamine (PGA) mutant. We compared the amount of biofilm produced by the parental strain and the isogenic mutants using static and dynamic systems. Compared to the findings for the biofilm of the parental or other strains, the biofilm of the O antigen and the PGA mutants was dramatically reduced, and it had less cell-associated PGA. Real-time PCR analyses revealed a significant reduction in the level of pgaA, cpxR, and cpxA mRNA in the biofilm cells of the O-antigen mutant compared to that in the biofilm cells of the parental strain. Specific binding between PGA and LPS was consistently detected by surface plasmon resonance, but the lack of O antigen did not abolish these interactions. In conclusion, the absence of the O antigen reduces the ability of A. pleuropneumoniae to form a biofilm, and this is associated with the reduced expression and production of PGA.A ctinobacillus pleuropneumoniae is a Gram-negative bacterium belonging to the Pasteurellaceae family and is the causative agent of porcine pleuropneumonia, a disease causing important economic losses to the swine industry worldwide (1). Several virulence factors of A. pleuropneumoniae have been identified. These factors include the Apx toxins, iron uptake systems, and surface polysaccharides. These polysaccharides are divided into three categories: the lipopolysaccharides (LPS), the capsular polysaccharides (CPS), and the poly-N-acetyl-D-glucosamine polymer (PGA) present in the biofilm matrix (2-4).LPS are large biomolecules composed of three well-defined regions: (i) lipid A, anchored in the outer membrane; (ii) the core oligosaccharide; and (iii) the O antigen, a polysaccharide consisting of repeating units. Altman and coworkers described the structure of the A. pleuropneumoniae serotype 1 O antigen to be branched tetrasaccharide repeating units composed of two Lrhamnopyranosyl residues, one D-glycopyranosyl residue, and one 2-acetamido-2-deoxy-D-glucose residue (5). The LPS inner core oligosaccharide is relatively conserved among A. pleuropneumoniae serotype 1, 2, 5a, and 5b strains and is composed of 2-keto-3-deoxyoctulosonic acid and heptose, whereas the outer core oligosaccharide is variable among serotypes and is generally made of a variable number of branching hexoses (6). The A. pleuropneumoniae LPS has been identified to b...

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A method for growing a biofilm under low shear at the air–liquid interface using the drip flow biofilm reactor

Cited by 193 publications

References 14 publications

Biofilm Models of Polymicrobial Infection

Biofilm Models of Polymicrobial Infection

Evidence for persisters in Staphylococcus epidermidis RP62a planktonic cultures and biofilms

Surface Polysaccharide Mutants Reveal that Absence of O Antigen Reduces Biofilm Formation of Actinobacillus pleuropneumoniae

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