scholarly journals Synergistic Antibacterial and Antibiofilm Activity of the MreB Inhibitor A22 Hydrochloride in Combination with Conventional Antibiotics against Pseudomonas aeruginosa and Escherichia coli Clinical Isolates

2021 ◽  
Vol 2021 ◽  
pp. 1-17
Author(s):  
Anastasia Kotzialampou ◽  
Efthymia Protonotariou ◽  
Lemonia Skoura ◽  
Afroditi Sivropoulou
Keyword(s):  
Escherichia Coli ◽  
Pseudomonas Aeruginosa ◽  
Inhibitory Concentration ◽  
Human Cells ◽  
Laser Scanning Microscopy ◽  
Clinical Isolates ◽  
Antibiofilm Activity ◽  
Conventional Antibiotics ◽  
Time Kill ◽  
Synergistic Combinations

In the era of antibiotic resistance, the bacterial cytoskeletal protein MreB is presented as a potential target for the development of novel antimicrobials. Combined treatments of clinical antibiotics with anti-MreB compounds may be promising candidates in combating the resistance crisis, but also in preserving the potency of many conventional drugs. This study aimed to evaluate the synergistic antibacterial and antibiofilm activities of the MreB inhibitor A22 hydrochloride in combination with various antibiotics. The minimum inhibitory concentration (MIC) values of the individual compounds were determined by the broth microdilution method against 66 clinical isolates of Gram-negative bacteria. Synergy was assessed by the checkerboard assay. The fractional inhibitory concentration index was calculated for each of the A22-antibiotic combination. Bactericidal activity of the combinations was evaluated by time-kill curve assays. The antibiofilm activity of the most synergistic combinations was determined by crystal violet stain, methyl thiazol tetrazolium assay, and confocal laser scanning microscopy analysis. The combined cytotoxic and hemolytic activity was also evaluated toward human cells. According to our results, Pseudomonas aeruginosa and Escherichia coli isolates were resistant to conventional antibiotics to varying degrees. A22 inhibited the bacterial growth in a dose-dependent manner with MIC values ranging between 2 and 64 μg/mL. In combination studies, synergism occurred most frequently with A22-ceftazidime and A22-meropemen against Pseudomonas aeruginosa and A22-cefoxitin and A22-azithromycin against Escherichia coli. No antagonism was observed. In time-kill studies, synergism was observed with all expected combinations. Synergistic combinations even at the lowest tested concentrations were able to inhibit biofilm formation and eradicate mature biofilms in both strains. Cytotoxic and hemolytic effects of the same combinations toward human cells were not observed. The findings of the present study support previous research regarding the use of MreB as a novel antibiotic target. The obtained data expand the existing knowledge about the antimicrobial and antibiofilm activity of the A22 inhibitor, and they indicate that A22 can serve as a leading compound for studying potential synergism between MreB inhibitors and antibiotics in the future.

scholarly journals In vitro antibiofilm activity of resveratrol against avian pathogenic Escherichia coli

2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Xiangchun Ruan ◽  
Xiaoling Deng ◽  
Meiling Tan ◽  
Chengbo Yu ◽  
Meishi Zhang ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Biofilm Formation ◽  
Inhibitory Concentration ◽  
Laser Scanning Microscopy ◽  
Antibiofilm Activity ◽  
Dependent Manner ◽  
Swarming Motility ◽  
Semisolid Medium ◽  
Broth Microdilution Method

Abstract Background Avian pathogenic Escherichia coli (APEC) strains cause infectious diseases in poultry. Resveratrol is extracted from Polygonum cuspidatum, Cassia tora Linn and Vitis vinifera, and displays good antimicrobial activity. The present study aimed to investigate the antibiofilm effect of resveratrol on APEC in vitro. The minimum inhibitory concentration (MIC) of resveratrol and the antibiotic florfenicol toward APEC were detected using the broth microdilution method. Then, the effect of resveratrol on swimming and swarming motility was investigated using a semisolid medium culture method. Subsequently, the minimum biofilm inhibitory concentration (MBIC) and the biofilm eradication rate were evaluated using crystal violet staining. Finally, the antibiofilm activity of resveratrol was observed using scanning electron microscopy (SEM). Meanwhile, the effects of florfenicol combined with resveratrol against biofilm formation by APEC were evaluated using optical microscopy (OM) and a confocal laser scanning microscopy (CLSM). Results The MICs of resveratrol and florfenicol toward APEC were 128 μg/mL and 64 μg/mL, respectively. The swimming and swarming motility abilities of APEC were inhibited in a resveratrol dose-dependent manner. Furthermore, resveratrol showed a significant inhibitory activity against APEC biofilm formation at concentrations above 1 μg/mL (p < 0.01). Meanwhile, the inhibitory effect of resveratrol at 32 μg/mL on biofilm formation was observed using SEM. The APEC biofilm was eradicated at 32 μg/mL of resveratrol combined with 64 μg/mL of florfenicol, which was observed using CLSM and OM. Florfenicol had a slight eradication effect of biofilm formation, whereas resveratrol had a strong biofilm eradication effect toward APEC. Conclusion Resveratrol displayed good antibiofilm activity against APEC in vitro, including inhibition of swimming and swarming motility, biofilm formation, and could eradicate the biofilm.

Confocal and SEM imaging to demonstrate food pathogen- biofilm biocontrol by pyocyanin from Pseudomonas aeruginosa BTRY1

2016 ◽  
Vol 6 (01) ◽  
pp. 5218
Author(s):  
Laxmi Mohandas ◽  
Anju T. R. ◽  
Sarita G. Bhat*
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Biofilm Formation ◽  
Laser Scanning ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Antibiofilm Activity ◽  
Opportunistic Pathogens ◽  
Redox Active ◽  
Sem Imaging ◽  
The Impact

An assortment of redox-active phenazine compounds like pyocyanin with their characteristic blue-green colour are synthesized by Pseudomonas aeruginosa, Gram-negative opportunistic pathogens, which are also considered one of the most commercially valuable microorganisms. In this study, pyocyanin from Pseudomonas aeruginosa BTRY1 from food sample was assessed for its antibiofilm activity by micro titer plate assay against strong biofilm producers belonging to the genera Bacillus, Staphylococcus, Brevibacterium and Micrococcus. Pyocyanin inhibited biofilm activity in very minute concentrations. This was also confirmed by Scanning Electron Microscopy (SEM) and Confocal Laser Scanning Microscopy (CLSM). Both SEM and CLSM helped to visualize the biocontrol of biofilm formation by eight pathogens. The imaging and quantification by CLSM also established the impact of pyocyanin on biofilm-biocontrol mainly in the food industry.

scholarly journals Cephalosporin nitric oxide-donor prodrug DEA-C3D disperses biofilms formed by clinical cystic fibrosis isolates of Pseudomonas aeruginosa

2019 ◽  
Vol 75 (1) ◽  
pp. 117-125 ◽  
Author(s):  
Odel Soren ◽  
Ardeshir Rineh ◽  
Diogo G Silva ◽  
Yuming Cai ◽  
Robert P Howlin ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Cystic Fibrosis ◽  
Pseudomonas Aeruginosa ◽  
Laser Scanning ◽  
Laser Scanning Microscopy ◽  
Clinical Isolates ◽  
Nitric Oxide Donor ◽  
Confocal Laser ◽  
Broth Microdilution Method

Abstract Objectives The cephalosporin nitric oxide (NO)-donor prodrug DEA-C3D (‘DiEthylAmin-Cephalosporin-3′-Diazeniumdiolate’) has been shown to initiate the dispersal of biofilms formed by the Pseudomonas aeruginosa laboratory strain PAO1. In this study, we investigated whether DEA-C3D disperses biofilms formed by clinical cystic fibrosis (CF) isolates of P. aeruginosa and its effect in combination with two antipseudomonal antibiotics, tobramycin and colistin, in vitro. Methods β-Lactamase-triggered release of NO from DEA-C3D was confirmed using a gas-phase chemiluminescence detector. MICs for P. aeruginosa clinical isolates were determined using the broth microdilution method. A crystal violet staining technique and confocal laser scanning microscopy were used to evaluate the effects of DEA-C3D on P. aeruginosa biofilms alone and in combination with tobramycin and colistin. Results DEA-C3D was confirmed to selectively release NO in response to contact with bacterial β-lactamase. Despite lacking direct, cephalosporin/β-lactam-based antibacterial activity, DEA-C3D was able to disperse biofilms formed by three P. aeruginosa clinical isolates. Confocal microscopy revealed that DEA-C3D in combination with tobramycin produces similar reductions in biofilm to DEA-C3D alone, whereas the combination with colistin causes near complete eradication of P. aeruginosa biofilms in vitro. Conclusions DEA-C3D is effective in dispersing biofilms formed by multiple clinical isolates of P. aeruginosa and could hold promise as a new adjunctive therapy to patients with CF.

scholarly journals The Antibiofilm Effect of a Medical Device Containing TIAB on Microorganisms Associated with Surgical Site Infection

Molecules ◽  
2019 ◽  
Vol 24 (12) ◽  
pp. 2280 ◽  
Author(s):  
Valentina Puca ◽  
Tonino Traini ◽  
Simone Guarnieri ◽  
Simone Carradori ◽  
Francesca Sisto ◽  
...  
Keyword(s):  
Medical Device ◽  
Biofilm Formation ◽  
Inhibitory Concentration ◽  
Laser Scanning Microscopy ◽  
Test Method ◽  
Antibiofilm Activity ◽  
E Coli ◽  
Post Surgery ◽  
Agar Diffusion Test ◽  
Surgical Sutures

Surgical site infections (SSIs) represent the most common nosocomial infections, and surgical sutures are optimal surfaces for bacterial adhesion and biofilm formation. Staphylococcus spp., Enterococcus spp., and Escherichia coli are the most commonly isolated microorganisms. The aim of this research was to evaluate the antibiofilm activity of a medical device (MD) containing TIAB, which is a silver-nanotech patented product. The antibacterial effect was evaluated against Staphylococcus aureus ATCC 29213, Enterococcus faecalis ATCC 29212, and E. coli ATCC 25922 by assessing the minimum inhibitory concentration (MIC) by the Alamar Blue® (AB) assay. The antibiofilm effect was determined by evaluation of the minimum biofilm inhibitory concentration (MBIC) and colony-forming unit (CFU) count. Subsequently, the MD was applied on sutures exposed to the bacterial species. The antimicrobial and antibiofilm effects were evaluated by the agar diffusion test method, confocal laser scanning microscopy (CLSM), and scanning electron microscopy (SEM). The MIC was determined for S. aureus and E. faecalis at 2 mg/mL, while the MBIC was 1.5 mg/mL for S. aureus and 1 mg/mL for E. faecalis. The formation of an inhibition zone around three different treated sutures confirmed the antimicrobial activity, while the SEM and CLSM analysis performed on the MD-treated sutures underlined the presence of a few adhesive cells, which were for the most part dead. The MD showed antimicrobial and antibiofilm activities versus S. aureus and E. faecalis, but a lower efficacy against E. coli. Surgical sutures coated with the MD have the potential to reduce SSIs as well as the risk of biofilm formation post-surgery.

scholarly journals In VitroAntibacterial and Antibiofilm Activities of Chlorogenic Acid against Clinical Isolates ofStenotrophomonas maltophiliaincluding the Trimethoprim/Sulfamethoxazole Resistant Strain

2013 ◽  
Vol 2013 ◽  
pp. 1-7 ◽  
Author(s):  
Arunkumar Karunanidhi ◽  
Renjan Thomas ◽  
Alex van Belkum ◽  
Vasanthakumari Neela
Keyword(s):  
Chlorogenic Acid ◽  
Inhibition Zone ◽  
Clinical Isolates ◽  
Antibiofilm Activity ◽  
Log Reduction ◽  
Fold Reduction ◽  
Viable Bacteria ◽  
Study Support ◽  
Time Kill

Thein vitroantibacterial and antibiofilm activity of chlorogenic acid against clinical isolates ofStenotrophomonas maltophiliawas investigated through disk diffusion, minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), time-kill and biofilm assays. A total of 9 clinicalS. maltophiliaisolates including one isolate resistant to trimethoprim/sulfamethoxazole (TMP/SMX) were tested. The inhibition zone sizes for the isolates ranged from 17 to 29 mm, while the MIC and MBC values ranged from 8 to 16 μg mL−1and 16 to 32 μg mL−1. Chlorogenic acid appeared to be strongly bactericidal at 4x MIC, with a 2-log reduction in viable bacteria at 10 h.In vitroantibiofilm testing showed a 4-fold reduction in biofilm viability at 4x MIC compared to 1x MIC values (0.085<0.397A 490 nm) of chlorogenic acid. The data from this study support the notion that the chlorogenic acid has promisingin vitroantibacterial and antibiofilm activities againstS. maltophilia.

Serum Inhibitory and Bactericidal Activity of Ciprofloxacin following Intravenous Administration

DICP ◽  
1989 ◽  
Vol 23 (6) ◽  
pp. 456-460
Author(s):  
Michael N. Dudley ◽  
Hilary D. Mandler ◽  
Kenneth H. Mayer ◽  
Stephen H. Zinner
Keyword(s):  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Pseudomonas Aeruginosa ◽  
Minimum Inhibitory Concentration ◽  
Bactericidal Activity ◽  
Resistant Strain ◽  
Inhibitory Concentration ◽  
Minimum Bactericidal Concentration ◽  
E Coli ◽  
Dose Levels

Serum inhibitory and bactericidal titers were measured in nine healthy volunteers following single iv doses of ciprofloxacin 100, 150, and 200 mg. The median peak serum bactericidal titer (5 minutes following completion of a 30-minute infusion) against two highly susceptible strains of Escherichia coli ranged between 1:64 and 1:1024 and titers exceeded 1:8 for six hours for all dose levels. The bactericidal titers against two strains of Pseudomonas aeruginosa and a methicillin-resistant strain of Staphylococcus aureus were considerably lower, the median peak being 1:2 at all dose levels. Measured inhibitory and bactericidal titers at five minutes and one hour postinfusion were significantly greater than those predicted (measured serum ciprofloxacin concentration to minimum inhibitory concentration [MIC] or minimum bactericidal concentration [MBC]) for only one strain of E. coli. Intravenous doses of ciprofloxacin 100–200 mg produce high and sustained serum bactericidal titers against highly susceptible bacteria; considerably lower levels of activity are seen against bacteria having higher MICs and MBCs but still considered susceptible to the drug.

scholarly journals Synergy of gatifloxacin with cefoperazone and cefoperazone–sulbactam against resistant strains of Pseudomonas aeruginosa

2008 ◽  
Vol 57 (12) ◽  
pp. 1514-1517 ◽  
Author(s):  
N. Sivagurunathan ◽  
S. Krishnan ◽  
J. Venkat Rao ◽  
Anantha Naik Nagappa ◽  
V. M. Subrahmanyam ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Clinical Isolates ◽  
Significant Difference ◽  
Resistant Strains ◽  
Time Kill

Chequerboard and time–kill methods were used to compare the in vitro efficacies of the combinations gatifloxacin (GAT) with cefoperazone (CFP) and GAT with cefoperazone–sulbactam (CFP-SUL) against 58 clinical isolates of Pseudomonas aeruginosa. The combinations GAT+CFP and GAT+CFP-SUL were shown to be synergistic for 36.2 and 58.6 % of isolates tested, respectively, using the chequerboard method. Time–kill studies with 11 strains showed synergy in 54.5 % for the GAT+CFP combination and 72.7 % for the GAT+CFP-SUL combination. The agreement between these two methods was found to be 72–81 %. There was a significant difference in synergy between the two combinations tested (P=0.011).

scholarly journals Evaluation of the antibacterial and modulatory activities of ethanolic excract of Calotropis procera (Aiton) W.T. Aiton against multiresistant bacterial strains

2021 ◽  
pp. 205-209
Author(s):  
Yuri Geraldo ◽  
Livia Leandro ◽  
Ana Raquel Silva ◽  
Fábia Campina ◽  
Ana Carolina Araújo ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Pseudomonas Aeruginosa ◽  
Inhibitory Concentration ◽  
Ethanolic Extract ◽  
Calotropis Procera ◽  
Subinhibitory Concentration ◽  
Bacterial Strains ◽  
Resistant Species ◽  
Microdilution Method

The objective of this study was to evaluate the antibacterial and modulatory activities of ethanolic extract of Calotropis procera (Aiton) W.T. Aiton against resistant species. By microdilution method, the minimum inhibitory concentration (MIC) of the extract and modulation of the subinhibitory concentration MIC/8 to norfloxacine, gentamicin and imipenem against Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa. There was obtained 512 μg/mL to Pseudomonas aeruginosa. To Staphylococcus aureus, modulation showed synergism to norfloxacin and gentamicin, with imipenem against Pseudomonas aeruginosa and gentamicin against Escherichia coli. Based on these results, more studies are needed to test the antibacterial activity of the extract. El objetivo de este estudio fue evaluar la actividad antibacteriana y moduladora del extracto etanólico de Calotropis procera (Aiton) W.T. Aiton contra cepas multirresistentes de bacterias. Por el método de microdilución, fueron definidas la concentración inhibidora mínima (MIC) del extracto y la modulación con la concentración inhibidora CIM / 8 del extracto con norfloxacina, gentamicina e imipenem contra Staphylococcus aureus, Escherichia coli y Pseudomonas aeruginosa. Se obtuvo 512 μg/mL para Pseudomonas aeruginosa. Se descubrió sinergismo en el caso de Staphylococcus aureus, en la modulación con norfloxacina y gentamicina, mientras que con imipenem frente a Pseudomonas aeruginosa y con gentamicina para Escherichia coli. Con base en estos resultados, se necesitan más estudios para probar la actividad antibacteriana del extracto.

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