Real-time monitoring of Pseudomonas aeruginosa biofilm formation on endotracheal tubes in vitro

Introduction. Indwelling medical devices such as endotracheal tubes (ETTs), urinary catheters, vascular access devices, tracheostomies and feeding tubes are often associated with hospital-acquired infections. Bacterial biofilm formed on the ETTs in intubated patients is a significant risk factor associated with ventilator-associated pneumonia. Pseudomonas aeruginosa is one of the four frequently encountered bacteria responsible for causing pneumonia, and the biofilm formation on ETTs. However, understanding of biofilm formation on ETT and interventions to prevent biofilm remains lagging. The ability to sense and adapt to external cues contributes to their success. Thus, the biofilm formation is likely to be influenced by the two-component systems (TCSs) that are composed of a membrane-associated sensor kinase and an intracellular response regulator. Aim. This study aims to establish an in vitro method to analyse the P. aeruginosa biofilm formation on ETTs, and identify the TCSs that contribute to this process. Methodology. In total, 112 P. aeruginosa PA14 TCS mutants were tested for their ability to form biofilm on ETTs, their effect on quorum sensing (QS) and motility. Results. Out of 112 TCS mutants studied, 56 had altered biofilm biomass on ETTs. Although the biofilm formation on ETTs is QS-dependent, none of the 56 loci controlled quorum signal. Of these, 18 novel TCSs specific to ETT biofilm were identified, namely, AauS, AgtS, ColR, CopS, CprR, NasT, KdpD, ParS, PmrB, PprA, PvrS, RcsC, PA14_11120, PA14_32580, PA14_45880, PA14_49420, PA14_52240, PA14_70790. The set of 56 included the GacS network, TCS proteins involved in fimbriae synthesis, TCS proteins involved in antimicrobial peptide resistance, and surface-sensing. Additionally, several of the TCS-encoding genes involved in biofilm formation on ETTs were found to be linked to flagellum-dependent swimming motility. Conclusions. Our study established an in vitro method for studying P. aeruginosa biofilm formation on the ETT surfaces. We also identified novel ETT-specific TCSs that could serve as targets to prevent biofilm formation on indwelling devices frequently used in clinical settings.

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Bioactivity of Phenolic Compounds Extracted from Strawberry against Formation of Pseudomonas aeruginosa Biofilm

International Journal of Pharmaceutical Quality Assurance ◽

10.25258/ijpqa.10.1.27 ◽

2019 ◽

Vol 10 (01) ◽

Author(s):

Baydaa Hussein ◽

Zainab A. Aldhaher ◽

Shahrazad Najem Abdu-Allah ◽

Adel Hamdan

Keyword(s):

Pseudomonas Aeruginosa ◽

Phenolic Compounds ◽

Biofilm Formation ◽

Opportunistic Infections ◽

Formation Rate ◽

Hydrocarbon Chain ◽

Health Concern ◽

Gas Chromatography Mass Spectrometry ◽

Phenolic Contents

Background: Biofilm is a bacterial way of life prevalent in the world of microbes; in addition to that it is a source of alarm in the field of health concern. Pseudomonas aeruginosa is a pathogenic bacterium responsible for all opportunistic infections such as chronic and severe. Aim of this study: This paper aims to provide an overview of the promotion of isolates to produce a biofilm in vitro under special circumstances, to expose certain antibiotics to produce phenotypic evaluation of biofilm bacteria. Methods and Materials: Three diverse ways were used to inhibited biofilm formation of P.aeruginosa by effect of phenolic compounds extracts from strawberries. Isolates produced biofilm on agar MacConkey under certain circumstances. Results: The results showed that all isolates were resistant to antibiotics except sensitive to azithromycin (AZM, 15μg), and in this study was conducted on three ways to detect the biofilm produced, has been detected by the biofilm like Tissue culture plate (TCP), Tube method (TM), Congo Red Agar (CRA). These methods gave a clear result of these isolates under study. Active compounds were analyzed in both extracts by Gas Chromatography-mass Spectrometry which indicate High molecular weight compound with a long hydrocarbon chain. Conclusion: Phenolic compounds could behave as bioactive material and can be useful to be used in pharmaceutical synthesis. Phenolic contents which found in leaves and fruits extracts of strawberries shows antibacterial activity against all strains tested by the ability to reduce the production of biofilm formation rate.

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An In Vitro Coumarin-Antibiotic Combination Treatment of Pseudomonas aeruginosa Biofilms

Natural Product Communications ◽

10.1177/1934578x20987744 ◽

2021 ◽

Vol 16 (1) ◽

pp. 1934578X2098774

Author(s):

Jinpeng Zou ◽

Yang Liu ◽

Ruiwei Guo ◽

Yu Tang ◽

Zhengrong Shi ◽

...

Keyword(s):

Pseudomonas Aeruginosa ◽

Drug Resistance ◽

Combination Treatment ◽

Crude Extract ◽

Biofilm Formation ◽

Bacterial Resistance ◽

Antibacterial Properties ◽

Biofilm Growth ◽

Antibiotic Combination

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.

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Assessment of in vivo versus in vitro biofilm formation of clinical methicillin-resistant Staphylococcus aureus isolates from endotracheal tubes

Scientific Reports ◽

10.1038/s41598-018-30494-7 ◽

2018 ◽

Vol 8 (1) ◽

Cited By ~ 8

Author(s):

Laia Fernández-Barat ◽

Soumaya Ben-Aicha ◽

Anna Motos ◽

Jordi Vila ◽

Francesc Marco ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Biofilm Formation ◽

Methicillin Resistant Staphylococcus Aureus ◽

Methicillin Resistant ◽

Endotracheal Tubes

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Real time monitoring of peptide delivery in vitro using high payload pH responsive nanogels

Polymer Chemistry ◽

10.1039/c9py01120j ◽

2020 ◽

Vol 11 (2) ◽

pp. 425-432 ◽

Cited By ~ 2

Author(s):

Shegufta Farazi ◽

Fan Chen ◽

Henry Foster ◽

Raelene Boquiren ◽

Shelli R. McAlpine ◽

...

Keyword(s):

Real Time ◽

Intracellular Delivery ◽

Peptide Delivery ◽

High Loading ◽

Loading Capacity ◽

Ph Responsive ◽

Real Time Monitoring ◽

High Payload

A pH responsive pMAA nanogel that demonstrates high loading capacity and rapid intracellular delivery of hydrophilic peptides.

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Facile solvothermal synthesis of ultrathin spinel ZnMn2O4 nanospheres: An efficient electrocatalyst for in vivo and in vitro real time monitoring of H2O2

Journal of Electroanalytical Chemistry ◽

10.1016/j.jelechem.2021.115674 ◽

2021 ◽

Vol 900 ◽

pp. 115674

Author(s):

Muthaiah Annalakshmi ◽

Sakthivel Kumaravel ◽

T.S.T. Balamurugan ◽

Shen-Ming Chen ◽

Ju-Liang He

Keyword(s):

Real Time ◽

Solvothermal Synthesis ◽

Real Time Monitoring

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Non-invasive, real-time reporting drug release in vitro and in vivo

Chemical Communications ◽

10.1039/c4cc09920f ◽

2015 ◽

Vol 51 (32) ◽

pp. 6948-6951 ◽

Cited By ~ 31

Author(s):

Yanfeng Zhang ◽

Qian Yin ◽

Jonathan Yen ◽

Joanne Li ◽

Hanze Ying ◽

...

Keyword(s):

Drug Release ◽

Real Time ◽

Near Infrared ◽

Reporting System ◽

Real Time Monitoring ◽

Near Infrared Fluorescence ◽

Non Invasive ◽

Fluorescence Dye

Anin vitroandin vivodrug-reporting system is developed for real-time monitoring of drug release via the analysis of the concurrently released near-infrared fluorescence dye.

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Effect of sub-inhibitory concentrations of cefepime on biofilm formation by Pseudomonas aeruginosa

Canadian Journal of Microbiology ◽

10.1139/cjm-2021-0229 ◽

2021 ◽

pp. 1-8

Author(s):

Soheir A.A. Hagras ◽

Alaa El-Dien M.S. Hosny ◽

Omneya M. Helmy ◽

Mounir M. Salem-Bekhit ◽

Faiyaz Shakeel ◽

...

Keyword(s):

Pseudomonas Aeruginosa ◽

Biofilm Formation ◽

Clinical Isolates ◽

Polymeric Chain ◽

Rt Pcr ◽

Chain Reaction ◽

Protein Encoding ◽

Encoding Gene ◽

Fimbrial Protein

This study investigated the effect of cefepime at sub-minimum inhibitory concentrations (sub-MICs) on in vitro biofilm formation (BF) by clinical isolates of Pseudomonas aeruginosa. The effect of cefepime at sub-MIC levels (½–1/256 MIC) on in vitro BF by six clinical isolates of P. aeruginosa was phenotypically assessed following 24 and 48 h of challenge using the tissue culture plate (TCP) assay. Quantitative real-time polymeric chain reaction (qRT-PCR) was employed to observe the change in expression of three biofilm-related genes, namely, a protease-encoding gene (lasA), fimbrial protein-encoding gene (cupA1), and alginate-encoding gene (algC), in a weak biofilm-producing strain of P. aeruginosa following 24 and 48 h of challenge with sub-MICs of cefepime. The BF morphology in response to cefepime was imaged using scanning electron microscopy (SEM). The TCP assay showed strain-, time-, and concentration-dependent changes in in vitro BF in P. aeruginosa following challenge with sub-MICs of cefepime, with a profound increase in strains with inherently no or weak biofilm-producing ability. RT-PCR revealed time-dependent upregulation in the expression of the investigated genes following challenge with ½ and ¼ MIC levels, as confirmed by SEM. Cefepime at sub-MICs could upregulate the expression of BF-related genes and enhance BF by P. aeruginosa clinical isolates.

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NIR-Responsive Lysosomotropic Phototrigger: ʽAIE + ESIPTʼ Active Naphthalene Based Single Component Photoresponsive Nanocarrier with Two-Photon Uncaging and Real-Time Monitoring Ability

10.33774/chemrxiv-2021-nftrb ◽

2021 ◽

Author(s):

Biswajit Roy ◽

Rakesh Mengji ◽

Samrat Roy ◽

Bipul Pal ◽

Avijit Jana ◽

...

Keyword(s):

Drug Delivery ◽

Drug Release ◽

Real Time ◽

Near Infrared ◽

Single Component ◽

Photon Absorption ◽

Real Time Monitoring ◽

Two Photon ◽

Nir Light

In recent times, organelle-targeted drug delivery systems gained tremendous attention due to the site specific delivery of active drug molecules resulting in enhanced bioefficacy. In this context, the phototriggered drug delivery system (DDS) for releasing an active molecule is superior as it provides spatial and temporal control over the release. So far, near infrared (NIR) light responsive organelle targeted DDS has not yet been developed. Hence, we introduced a two-photon NIR-light responsive lysosome targeted ʽAIE + ESIPTʼ active single component DDS based on naphthalene chromophore. The Two-photon absorption cross-section of our DDS is 142 GM at 850 nm. The DDS was converted into pure organic nanoparticles for biological applications. Our nano-DDS is capable of selective targeting, AIE-luminogenic imaging, and drug release within the lysosome. In vitro studies using cancerous cell lines showed that our single component photoresponsive nanocarrier exhibited enhanced cytotoxicity and real-time monitoring ability of the drug release.

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Inhibitory effect of propafenone derivatives on pseudomonas aeruginosa biofilm and pyocyanin production

Srpski arhiv za celokupno lekarstvo ◽

10.2298/sarh180727102p ◽

2020 ◽

Vol 148 (3-4) ◽

pp. 196-202

Author(s):

Snjezana Petrovic ◽

Jasmina Basic ◽

Zoran Mandinic ◽

Dragana Bozic ◽

Marina Milenkovic ◽

...

Keyword(s):

Pseudomonas Aeruginosa ◽

Biofilm Formation ◽

Laboratory Strain ◽

Inhibitory Effect ◽

Negative Control ◽

Clinical Strains ◽

Control Value ◽

Essential Components ◽

Pyocyanin Production

Introduction/Objective. Biofilm and pyocyanin production are essential components of Pseudomonas aeruginosa virulence and antibiotic resistance. Our objective was to examine inhibitory effect of synthetized propafenone derivatives 3-(2-Fluorophenyl)- 1-(2- (2-hydroxy-3-propylamino-propoxy)-phenyl)-propan-1-one hydrochloride (5OF) and3-(2- Trifluoromethyl-phenyl)-1-(2-(2-hydroxy-3-propylamino-propoxy)-phenyl)-propan-1-one hydrochloride (5CF3) on biofilm and pyocyanin in Pseudomonas aeruginosa clinical strains. Methods. Effects were tested on nine clinical isolates and one control laboratory strain of P. aeruginosa. In vitro analysis of biofilm growing was performed by incubating bacteria (0.5 McFarland) with 5OF and 5CF3 (500?31.2 ?g/ml) and measuring optical density (OD) at 570 nm. Bacteria in medium without compounds were positive control. Blank medium (an uninoculated medium without test compounds) was used as negative control. Pyocyanin production was estimated by OD at 520 nm, after bacteria incubated with 5CF3 and 5OF (250 and 500 ?g/ml), treated with chloroform, and chloroform layer mixed with HCl. Results. A total of 500 ?g/ml of 5OF and 5CF3 completely inhibited biofilm formation in 10/10 and 4/10 strains, respectively. A total of 250 ?g/ml of 5OF and 5CF3 strongly inhibited biofilm formation in 7/10 strains, while inhibition with 125 ?g/ml of 5OF and 5CF3 was moderate. Lower concentrations had almost no effect on biofilm production. Pyocyanin production was reduced to less than 40% of the control value in 6/9, and less than 50% of the control in 7/9 strains with 500 ?g/ml of 5OF and 5CF3, respectively. At 250 ?g/ml 5OF and 5CF3, most strains had pyocyanin production above 50% of the control value. Conclusion. Synthetized propafenone derivatives, 5OF and 5CF3, inhibited biofilms and pyocyanin production of Pseudomonas aeruginosa clinical strains. Presented results suggest that propafenone derivatives are potential lead-compounds for synthesis of novel antipseudomonal drugs.

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