Differential surface competition and biofilm invasion strategies of Pseudomonas aeruginosa PA14 and PAO1

2021 ◽  
Author(s):  
Swetha Kassety ◽  
Stefan Katharios-Lanwermeyer ◽  
George A. O’Toole ◽  
Carey D. Nadell

Pseudomonas aeruginosa strains PA14 and PAO1 are among the two best characterized model organisms used to study the mechanisms of biofilm formation, while also representing two distinct lineages of P. aeruginosa . Previous work has shown that PA14 and PAO1 use different strategies for surface colonization; they also have different extracellular matrix composition and different propensities to disperse from biofilms back into the planktonic phase surrounding them. We expand on this work here by exploring the consequences of these different biofilm production strategies during direct competition. Using differentially labeled strains and microfluidic culture methods, we show that PAO1 can outcompete PA14 in direct competition during early colonization and subsequent biofilm growth, that they can do so in constant and perturbed environments, and that this advantage is specific to biofilm growth and requires production of the Psl polysaccharide. In contrast, the P. aeruginosa PA14 is better able to invade pre-formed biofilms and is more inclined to remain surface-associated under starvation conditions. These data together suggest that while P. aeruginosa PAO1 and PA14 are both able to effectively colonize surfaces, they do so in different ways that are advantageous under different environmental settings. Importance Recent studies indicate that P. aeruginosa PAO1 and PA14 use distinct strategies to initiate biofilm formation. We investigated whether their respective colonization and matrix secretion strategies impact their ability to compete under different biofilm-forming regimes. Our work shows that these different strategies do indeed impact how these strains fair in direct competition: PAO1 dominates during colonization of a naïve surface, while PA14 is more effective in colonizing a pre-formed biofilm. These data suggest that even for very similar microbes there can be distinct strategies to successfully colonize and persist on surfaces during the biofilm life cycle.

2021 ◽  
Author(s):  
Stefan Katharios-Lanwermeyer ◽  
Swetha Kasetty ◽  
Carey D Nadell ◽  
George O'Toole

Pseudomonas aeruginosa strains PA14 and PAO1 are among the two best characterized model organisms used to study the mechanisms of biofilm formation, while also representing two distinct lineages of P. aeruginosa. Our previous work showed that P. aeruginosa PA14 and PAO1 use distinct strategies to initiate biofilm growth. Using differentially-labeled strains and microfluidic devices, we show that PAO1 can outcompete PA14 in a head-to-head competition during early colonization of a surface, can do so in constant and perturbed environments, that this advantage is specific to biofilm growth and requires production of the Psl polysaccharide. In contrast, the P. aeruginosa PA14 exhibits a competitive fitness advantage when invading a pre-formed biofilm and is better able to tolerate starvation than PAO1 in the biofilm context. These data support the model that while P. aeruginosa PAO1 and PA14 are both able to effectively colonize surfaces, these strains use distinct strategies that are advantageous under different environmental settings.


2021 ◽  
Vol 16 (1) ◽  
pp. 1934578X2098774
Author(s):  
Jinpeng Zou ◽  
Yang Liu ◽  
Ruiwei Guo ◽  
Yu Tang ◽  
Zhengrong Shi ◽  
...  

The drug resistance of Pseudomonas aeruginosa is a worldwide problem due to its great threat to human health. A crude extract of Angelica dahurica has been proved to have antibacterial properties, which suggested that it may be able to inhibit the biofilm formation of P. aeruginosa; initial exploration had shown that the crude extract could inhibit the growth of P. aeruginosa effectively. After the adaptive dose of coumarin was confirmed to be a potential treatment for the bacteria’s drug resistance, “coumarin-antibiotic combination treatments” (3 coumarins—simple coumarin, imperatorin, and isoimperatorin—combined with 2 antibiotics—ampicillin and ceftazidime) were examined to determine their capability to inhibit P. aeruginosa. The final results showed that (1) coumarin with either ampicillin or ceftazidime significantly inhibited the biofilm formation of P. aeruginosa; (2) coumarin could directly destroy mature biofilms; and (3) the combination treatment can synergistically enhance the inhibition of biofilm formation, which could significantly reduce the usage of antibiotics and bacterial resistance. To sum up, a coumarin-antibiotic combination treatment may be a potential way to inhibit the biofilm growth of P. aeruginosa and provides a reference for antibiotic resistance treatment.


2006 ◽  
Vol 69 (10) ◽  
pp. 2411-2416 ◽  
Author(s):  
L. LAGACÉ ◽  
M. JACQUES ◽  
A. A. MAFU ◽  
D. ROY

The susceptibility of planktonic and biofilm cells of Pseudomonas marginalis toward four commonly used biocides at different temperatures (15 and 30°C) and biofilm growth times (24 and 48 h) was assessed. Using the MBEC biofilm device, biofilm production in maple sap was shown to be highly reproducible for each set of conditions tested. Biofilm formation was influenced by growth temperature and time. A temperature of 15°C and incubation time of 24 h yielded fewer CFU per peg and showed fewer adhered cells and typical biofilm structures, based on scanning electron microscopy observations as compared with other conditions. Minimal biofilm eradication concentration values for P. marginalis were significantly greater (P < 0.001) than were MBCs for planktonic cells and for every biocide tested, with the exception of minimal biofilm eradication concentration values for peracetic acid at 15°C and 24 h. Sodium hypochlorite and peracetic acid sanitizers were able to eliminate P. marginalis biofilms at lower concentrations as compared with hydrogen peroxide– and quaternary ammonium– based sanitizers (P < 0.001). According to the results obtained, sodium hypochlorite and peracetic acid sanitizers would be more appropriate for maple sap collection system sanitation.


2013 ◽  
Vol 57 (4) ◽  
pp. 1921-1925 ◽  
Author(s):  
Kenneth S. Brandenburg ◽  
Karien J. Rodriguez ◽  
Jonathan F. McAnulty ◽  
Christopher J. Murphy ◽  
Nicholas L. Abbott ◽  
...  

ABSTRACTBiofilm formation byPseudomonas aeruginosahas been implicated in the pathology of chronic wounds. Both thedandlisoforms of tryptophan inhibitedP. aeruginosabiofilm formation on tissue culture plates, with an equimolar ratio ofdandlisoforms producing the greatest inhibitory effect. Addition ofd-/l-tryptophan to existing biofilms inhibited further biofilm growth and caused partial biofilm disassembly. Tryptophan significantly increased swimming motility, which may be responsible in part for diminished biofilm formation byP. aeruginosa.


2020 ◽  
Vol 51 (5) ◽  
pp. 1413-1419
Author(s):  
Mahmood & et al.

The current study was designed to explore the association between the pigments production and biofilm construction in local Pseudomonas aeruginosa isolates. Out of 143 patients suffering from burns, urinary tract infections (UTI), respiratory tract infections and cystic fibrosis obtained from previous study by Mahmood (2015), twenty two isolates  (15.38%) were identified  from (11) hospitals in Iraq, splitted  into three provinces, Baghdad, Al-Anbar and Karbala for the duration of June 2017 to April 2018.  Characterization was carried out by using microscopical, morphological and biochemical methods which showed that all these isolates belong to P. aeruginosa.  Screening of   biofilm production isolates was carried out by using nutrient broth supplemented with glucose (0.25%) production medium which encourage this biofilm production. The percentage of pigmented isolates were collected from a total of 143 samples, 2.8% of the isolates from burns, 2.1% isolates from cystic fibrosis and 0.7% isolates from UTI.  Quantitative assays for biofilm formation were conducted using ELIZA technique. The results showed that all (22) isolates produced biofilm except one (B1 isolate). Biofilm quantities were varied from strong to medium production in comparison with control (0.0663). Statistical analysis results using Fischer's Exact test (p<0.05) were non-significant, therefore the pigment production has no association with biofilm formation for all of them.


2022 ◽  
Vol 13 (1) ◽  
pp. 88-92
Author(s):  
M Swapna ◽  
G Sumathi ◽  
M Anitha

Background: Pseudomonas aeruginosa is one of the most prevalent nosocomial pathogens that cause a life-threatening infection. One of the important characteristics of P. aeruginosa is biofilm formation which leads to antibiotic resistance. Aims and Objectives: The aim of the study was to study the antibiotic resistance pattern of P. aeruginosa isolates and correlation with their biofilm-production. Materials and Methods: A total of 87 P. aeruginosa isolates from different clinical specimens were processed and confirmed by conventional microbiological methods as per standard methodology. Antibiotic sensitivity testing was done for all isolates. Biofilm producing isolates were identified by the microtiter plate method (MTPM). Results: Of 87 P. aeruginosa isolates, majority were from pus 33 (38%), followed by urine 26 (30%), sputum 19 (22%), body fluids 7 (8%), and blood 2 (2%). Biofilm producing isolates showed more resistance in comparison to non-biofilm producers. The observed difference between biofilm formation for multidrug resistant and susceptible isolates was found to be statistically significant. Conclusion: MTPM method was an effective test for detection of biofilm formation and was also able to verify biofilm production by P. aeruginosa. This indicated a higher propensity among the clinical isolates of P. aeruginosa to form biofilm and revealed a positive correlation between biofilm formation and antibiotic resistance. This indicates the need for testing of even susceptible isolates for virulence factors such as biofilm production.


Author(s):  
Adithi R. Varadarajan ◽  
Raymond N. Allan ◽  
Jules D. P. Valentin ◽  
Olga E. Castañeda Ocampo ◽  
Vincent Somerville ◽  
...  

AbstractPseudomonas aeruginosa MPAO1 is the parental strain of the widely utilized transposon mutant collection for this important clinical pathogen. Here, we validate a model system to identify genes involved in biofilm growth and antibiotic resistance.Our model employs a genomics-driven workflow to assemble the complete MPAO1 genome, identify unique and conserved genes by comparative genomics with the PAO1 reference strain and missed genes by proteogenomics. Among over 200 unique MPAO1 genes, we identified six general essential genes that were overlooked when mapping public Tn-seq datasets against PAO1, including an antitoxin. Genomic data were integrated with phenotypic data from an experimental workflow using a user-friendly, soft lithography-based microfluidic flow chamber for biofilm growth. Experiments conducted across three laboratories delivered reproducible data on P. aeruginosa biofilms and validated both known and novel genes involved in biofilm growth and antibiotic resistance identified in screens of the mutant collection. Differential protein expression data from planktonic cells versus biofilm confirmed upregulation of candidates known to affect biofilm formation, of structural and secreted proteins of type six secretion systems, and provided proteogenomic evidence for some missed MPAO1 genes. This integrated, broadly applicable model promises to improve the mechanistic understanding of biofilm formation, antimicrobial tolerance and resistance evolution.


2020 ◽  
Vol 6 (1) ◽  
Author(s):  
Adithi R. Varadarajan ◽  
Raymond N. Allan ◽  
Jules D. P. Valentin ◽  
Olga E. Castañeda Ocampo ◽  
Vincent Somerville ◽  
...  

Abstract Pseudomonas aeruginosa MPAO1 is the parental strain of the widely utilized transposon mutant collection for this important clinical pathogen. Here, we validate a model system to identify genes involved in biofilm growth and biofilm-associated antibiotic resistance. Our model employs a genomics-driven workflow to assemble the complete MPAO1 genome, identify unique and conserved genes by comparative genomics with the PAO1 reference strain and genes missed within existing assemblies by proteogenomics. Among over 200 unique MPAO1 genes, we identified six general essential genes that were overlooked when mapping public Tn-seq data sets against PAO1, including an antitoxin. Genomic data were integrated with phenotypic data from an experimental workflow using a user-friendly, soft lithography-based microfluidic flow chamber for biofilm growth and a screen with the Tn-mutant library in microtiter plates. The screen identified hitherto unknown genes involved in biofilm growth and antibiotic resistance. Experiments conducted with the flow chamber across three laboratories delivered reproducible data on P. aeruginosa biofilms and validated the function of both known genes and genes identified in the Tn-mutant screens. Differential protein abundance data from planktonic cells versus biofilm confirmed the upregulation of candidates known to affect biofilm formation, of structural and secreted proteins of type VI secretion systems, and provided proteogenomic evidence for some missed MPAO1 genes. This integrated, broadly applicable model promises to improve the mechanistic understanding of biofilm formation, antimicrobial tolerance, and resistance evolution in biofilms.


2014 ◽  
Vol 2014 ◽  
pp. 1-5 ◽  
Author(s):  
R. Seixas ◽  
M. Gabriel ◽  
J. Machado ◽  
L. Tavares ◽  
F. Bernardo ◽  
...  

SalmonellaTyphimurium 1,4,[5],12:i:- is a major serovar responsible for human salmonellosis whose biofilm-forming ability, influenced by environmental conditions like those found in the gastrointestinal tract, is one of the main contributing factors to its ability to persist in the host and thus one of the main causes of chronic relapsing infections. Most studies to evaluate biofilm formation are performed in microtiter assays using standard media. However, no reports are available on the ability of this serovar to produce biofilm underin vitrosimulated gastrointestinal conditions which better correlate with the environment found in the gastrointestinal tract. To address this, a modified biofilm assay simulating intestinal fluid was conceived to assess the biofilm formation of 133SalmonellaTyphimurium 1,4,[5],12:i:- isolates with and without agitation and at three different time points (24 h, 48 h, and 72 h). The results were then compared to the existing microtiter method using conventional biofilm growth medium (Mueller Hinton Broth). Statistical analysis revealed significant differences in the results obtained between the three protocols used. The simulated human intestinal environment impaired biofilm production demonstrating that conditions like pH, agitation or the presence of enzymes can influence biofilm production. Therefore, results fromin vitrosimulation ofin vivoconditions may contribute to unravelling factors relating to biofilm formation and persistence in the context of the human host.


2020 ◽  
Author(s):  
Todd C. Chappell ◽  
Nikhil U. Nair

AbstractBiofilms are an emerging target for new therapeutics in the effort to address the continued increase in resistance and tolerance to traditional antimicrobials. In particular, the distinct nature of the biofilm growth state often means that traditional antimicrobials, developed to combat planktonic cells, are ineffective. Biofilm treatments are designed to both reduce pathogen load at an infection site and decrease the development of resistance by rendering the embedded organisms more susceptible to treatment at lower antimicrobial concentrations. In this work, we developed a new antimicrobial treatment modality by characterizing the natural capacity of two lactobacilli, L. plantarum and L. rhamnosus, to inhibit P. aeruginosa growth, biofilm formation, and biofilm viability. We further engineered these lactic acid bacteria (LAB) to secrete enzymes known to degrade P. aeruginosa biofilms and show that our best performing engineered LAB, secreting a pathogen-derived enzyme (PelAhyd), degrades up to 85 % of P. aeruginosa biofilm.


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