scholarly journals Antibiofilm Activity of Heather and Manuka Honeys and Antivirulence Potential of Some of Their Constituents on the DsbA1 Enzyme of Pseudomonas aeruginosa

Antibiotics ◽  
2020 ◽  
Vol 9 (12) ◽  
pp. 911
Author(s):  
Oscar Shirlaw ◽  
Zara Billah ◽  
Baraa Attar ◽  
Lisa Hughes ◽  
Rana M. Qasaymeh ◽  
...  

Heather honey was tested for its effect on the formation of biofilms by Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae, Enterococcus faecalis, Salmonella Enteriditis and Acinetobacter baumanii in comparison with Manuka honey. At 0.25 mg/mL, Heather honey inhibited biofilm formation in S. aureus, A. baumanii, E. coli, S. Enteriditis and P. aeruginosa, but promoted the growth of E. faecalis and K. pneumoniae biofilms. Manuka honey inhibited biofilm formation in K. pneumoniae, E. faecalis, and S. Enteriditis, A. baumanii, E. coli and P. aeruginosa, but promoted S. aureus biofilm formation. Molecular docking with Autodock Vina was performed to calculate the predictive binding affinities and ligand efficiencies of Manuka and Heather honey constituents for PaDsbA1, the main enzyme controlling the correct folding of virulence proteins in Pseudomonas aeruginosa. A number of constituents, including benzoic acid and methylglyoxal, present in Heather and/or Manuka honey, revealed high ligand efficiencies for the target enzyme. This helps support, to some extent, the decrease in P. aeruginosa biofilm formation observed for such honeys.

2016 ◽  
Vol 6 (01) ◽  
pp. 5218
Author(s):  
Laxmi Mohandas ◽  
Anju T. R. ◽  
Sarita G. Bhat*

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.


Molecules ◽  
2021 ◽  
Vol 26 (6) ◽  
pp. 1710
Author(s):  
Mahmoud Emam ◽  
Doaa R. Abdel-Haleem ◽  
Maha M. Salem ◽  
Lina Jamil M. Abdel-Hafez ◽  
Rasha R. Abdel Latif ◽  
...  

Infections associated with the emergence of multidrug resistance and mosquito-borne diseases have resulted in serious crises associated with high mortality and left behind a huge socioeconomic burden. The chemical investigation of Lavandulacoronopifolia aerial parts extract using HPLC–MS/MS led to the tentative identification of 46 compounds belonging to phenolic acids, flavonoids and their glycosides, and biflavonoids. The extract displayed larvicidal activity against Culex pipiens larvae (LC50 = 29.08 µg/mL at 72 h). It significantly inhibited cytochrome P-450 monooxygenase (CYP450), acetylcholinesterase (AChE), and carboxylesterase (CarE) enzymes with the comparable pattern to the control group, which could explain the mode of larvae toxification. The extract also inhibited the biofilm formation of Pseudomonas aeruginosa by 17–38% at different Minimum Inhibitory Concentrations (MICs) (0.5–0.125 mg/mL) while the activity was doubled when combined with ciprofloxacin (ratio = 1:1 v:v). In conclusion, the wild plant, L.coronopifolia, can be considered a promising natural source against resistant bacteria and infectious carriers.


2019 ◽  
Vol 30 (3) ◽  
pp. 252-258 ◽  
Author(s):  
Maurício Malheiros Badaró ◽  
Vanessa Maria Fagundes Leite-Fernandes ◽  
Luciano Trevisan Martin ◽  
Viviane de Cássia Oliveira ◽  
Evandro Watanabe ◽  
...  

Abstract The disadvantage of liners materials is the difficulty of biofilm control. It was compared an experimental dentifrice contained Ricinus communis, with commercials dentifrices as antibiofilm activity against microorganisms on denture liner. Six hundred specimens were distributed in 5 groups (n=18/ microorganism): water; experimental dentifrice; specific dentifrice for denture and two conventional dentifrices against C. albicans; C. glabrata; S. mutans; S. aureus; E. coli. Each group had a negative (n=5; without contamination) and positive control (n=15/ microorganism; without cleaning). The antibiofilm activity was evaluated by the method of biofilm formation in triplicate. The specimens were contaminated in a standard way and incubated. After that, manual brushing was performed (60 s), washed with PBS, immersed in liquid culture medium for resuspension and sowing in solid medium. The results (mean of triplicates) were expressed in CFU/mL. The data was submitted to Shapiro-Wilk, ANOVA and Tukey test (p<0.05). The specific dentifrice (1.27±1.20) was the most effective against S. mutans, followed by conventional (Trihydral, 3.13±0.88; Colgate, 2.16±2.02) and experimental (3.81±1.37) dentifrices, which were similar to each other (p=0.008). All of them were different from water (4.79±1.42). The specific (0.21±0.21) and experimental (0.36±0.25) dentifrices were similar against S. aureus, with a higher mean of CFU when compared to conventional (Colgate, 0.06±0.13), which was more efficient (p=0.000). For C. albicans, C. glabrata and E. coli, all dentifrices were similar to water (p=0.186). It was concluded, that the experimental dentifrice was effective against S. aureus and had not efficacy against Candida spp.; S. mutans; E. coli, as occurred with the commercials dentifrices.


2019 ◽  
Vol 2019 ◽  
pp. 1-12
Author(s):  
Edward Ntim Gasu ◽  
Hubert Senanu Ahor ◽  
Lawrence Sheringham Borquaye

Bacteria in biofilms are encased in an extracellular polymeric matrix that limits exposure of microbial cells to lethal doses of antimicrobial agents, leading to resistance. In Pseudomonas aeruginosa, biofilm formation is regulated by cell-to-cell communication, called quorum sensing. Quorum sensing facilitates a variety of bacterial physiological functions such as swarming motility and protease, pyoverdine, and pyocyanin productions. Peptide mix from the marine mollusc, Olivancillaria hiatula, has been studied for its antibiofilm activity against Pseudomonas aeruginosa. Microscopy and microtiter plate-based assays were used to evaluate biofilm inhibitory activities. Effect of the peptide mix on quorum sensing-mediated processes was also evaluated. Peptide mix proved to be a good antibiofilm agent, requiring less than 39 μg/mL to inhibit 50% biofilm formation. Micrographs obtained confirmed biofilm inhibition at 1/2 MIC whereas 2.5 mg/mL was required to degrade preformed biofilm. There was a marked attenuation in quorum sensing-mediated phenotypes as well. At 1/2 MIC of peptide, the expression of pyocyanin, pyoverdine, and protease was inhibited by 60%, 72%, and 54%, respectively. Additionally, swarming motility was repressed by peptide in a dose-dependent manner. These results suggest that the peptide mix from Olivancillaria hiatula probably inhibits biofilm formation by interfering with cell-to-cell communication in Pseudomonas aeruginosa.


Molecules ◽  
2019 ◽  
Vol 24 (12) ◽  
pp. 2280 ◽  
Author(s):  
Valentina Puca ◽  
Tonino Traini ◽  
Simone Guarnieri ◽  
Simone Carradori ◽  
Francesca Sisto ◽  
...  

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.


Author(s):  
Neda Fazeli ◽  
Akram Sadat Naeemi ◽  
Seyed Amir Hossein Jalali ◽  
Hojjatollah Zamani

Background: Staphylococcus aureus and Pseudomonas aeruginosa are important human bacterial pathogens, which are resistant to several antibiotics. One of the main causes of their resistance is the ability of biofilm formation. Objectives: The present study aimed to evaluate the antibacterial and antibiofilm activity of the extracts of Vibrio parahaemolyticus, V. alginolyticus, Pseudoalteromonas gelatinilytica, and Pseudoalteromonas piscicida isolated from sea anemone (Stichodactyla haddoni) against S. aureus and P. aeruginosa. Methods: Four isolated bacteria were identified using biochemical and molecular identification methods, and their extracts were obtained by mixing the cell-free supernatants from their old broth culture using ethyl acetate and methanol as the solvents. The agar well-diffusion and micro-dilution methods were also applied to determine the antibacterial activity, minimum bactericidal concentration (MBC), and minimum inhibitory concentration (MIC) of the extracts. The ability of the extracts to inhibit biofilm formation and disrupt the preformed biofilm of the pathogens was attained through crystal violet staining in 96-well microtiter plates. To determine the nature of the extracts, they were exposed to protease enzyme, and the antibiofilm activity was compared with the untreated extracts. Results: The extracts of the four isolated bacteria inhibited bacterial growth and biofilm formation and disrupted the preformed biofilm of S. aureus (MIC = BIC = 600 µg/mL) and P. aeruginosa (MIC = BIC = 300 µg/mL). In addition, the active compounds of the extracts with antibiofilm activities were mainly proteases. Conclusions: According to the results, V. parahaemolyticus, V. alginolyticus, P. gelatinilytica, and P. piscicida had antibacterial and antibiofilm potential against S. aureus and P. aeruginosa, and their extract could also be further analyzed as an alternative to antibiotics.


2020 ◽  
Vol 18 ◽  
Author(s):  
Sepideh Hassanzadeh ◽  
Sudabeh Ebrahimi ◽  
Sara Ganjloo ◽  
Saeid Amel Jamehdar ◽  
Samaneh Dolatabadi

Introduction: The biofilm formation by Pseudomonas aeruginosa seems to protect the bacteria from antibiotics since these entities are highly resistant to such antimicrobial agents. The aim of this study was to investigate the role of Lactobacillus salivarus, Lactobacillus plantarum supernatants and CuII Schiff base complex in eliminating planktonic cells and biofilm of P. aeruginosa. Methods: : One hundred specimens of blood, urine, cerebrospinal fluid, respiratory samples, and wound swabs were collected from patients attending three hospitals in Mashhad. All specimens were identified by biochemical tests. The susceptibility of the isolates to the conventional antibiotics were assessed using disk diffusion method. The biofilm formation ability of P. aeruginosa isolates was evaluated by crystal violet assay and confirmed using PCR. The anti-planktonic and anti-biofilm ability of L. salivarus, L. plantarum supernatants and CuII Schiff base complex was evaluated separately in P. aeruginosa isolates. Results and Conclusion: The highest and lowest resistance rates was detected in Cefazoline (95%) and cefepime (23%), respectively. The thickest biofilm was produced by 8% of P. aeruginosa isolates, 9% and 83% of the isolates were considered as moderate and weak biofilm producers, respectively. The rhlR and lasR genes was reported in 100% of the isolates, but algD gene was existence in 92% of them. Particularly, the CuII Schiff base complex could affect both planktonic and biofilm cells by the lowest concentration in comparison of probiotic supernatants. L. plantarum supernatant inhibited planktonic cells at a lower concentration than L. salivarius. Also, L. salivarius showed better antibiofilm activity than another probiotic in lower doses of supernatant. Unlike that these compounds have not completely eliminated biofilm cells, but only reduced the biofilm formation.Metal Schiff base complex and Lactobacillus supernatants is a potent antimicrobial agent against Pseudomonas aeruginosa biofilm cells.


2020 ◽  
Author(s):  
Anna Scherhag ◽  
Martina Rüger ◽  
Katrin Gerbracht ◽  
Jaqueline Rehner ◽  
Susanne Zehner ◽  
...  

&lt;p&gt;The molecule c-di-GMP is a bacterial second messenger that controls various processes such as motility or biofilm formation in bacteria [1]. To synthesize and degrade c-di-GMP, enzymes called diguanylate cyclases (DGC) containing a GGDEF-domain and phosphodiesterases (PDE) containing an EAL-domain or HD-GYP-domain are important [1, 2].&lt;em&gt; Pseudomonas aeruginosa&lt;/em&gt;, a model organism for biofilm formation and dispersion, encodes for 18 GGDEF, 5 EAL, 16 GGDEF / EAL, and 3 HD-GYP-domain-containing proteins [3].&lt;br /&gt;One of the GGDEF / EAL-containing proteins is NbdA. This protein also harbors an N-terminal membrane anchored MHYT-domain, that is predicted to be a sensor for NO, CO or O&lt;sub&gt;2&lt;/sub&gt; [4]. In this work, recombinant and affinity purified NbdA was tested for its PDE activity. Three different methods were used to measure the PDE activity of NbdA: a bis-pNPP-assay in which the conversion of the pseudosubstrate bis-pNPP into p-nitrophenol was detected spectroscopically, an HPLC-analysis of an enzymatic assay with the native substrate c-di-GMP, and a MANT-c-di-GMP-assay in which a fluorescently labeled form of the presumed substrate c-di-GMP was utilized.&lt;br /&gt;To establish these methods, the two known phosphodiesterases, PdeH from &lt;em&gt;Escherichia coli&lt;/em&gt; [5] and RocR from &lt;em&gt;P. aeruginosa&lt;/em&gt; [6], were also produced and tested. Subsequently, three variants of NbdA were investigated: the full-length version and two truncated versions of the protein. Activity was further assessed using functional complementation of an &lt;em&gt;E. coli&lt;/em&gt; phosphodiesterase deficient strain with full-length and truncated NbdA variants confirming PDE activity &lt;em&gt;in vivo&lt;/em&gt;.&lt;/p&gt; &lt;p&gt;&amp;#160;&lt;/p&gt; &lt;p&gt;&amp;#160;&lt;/p&gt; &lt;p&gt;[1] Hengge, R. (2009) Nature Rev. Microbiol. 7: 263-273.&lt;/p&gt; &lt;p&gt;[2] R&amp;#246;mling, U., Gomelsky, M., Galperin, M.Y. (2005). Mol. Microbiol. 57: 629&amp;#8211;639.&lt;/p&gt; &lt;p&gt;[3] Valentini, M., Filloux, A. (2016). J. Biol. Chem. 291: 12547&amp;#8211;12555.&lt;/p&gt; &lt;p&gt;[4] Galperin, M.Y., Gaidenko, T.A., Mulkidjanian, A.Y., Nakano, M., und Price, C.W. (2001). FEMS Microbiol. Lett. 205, 17&amp;#8211;23.&lt;/p&gt; &lt;p&gt;[5] Pesavento, C., Becker, G., Sommerfeldt, N., Possling, A., Tschowri, N., Mehlis, A., Hengge, R. (2008). Genes Dev. 22: 2434&amp;#8211;2446.&lt;/p&gt; &lt;p&gt;[6] Chen et al. (2012) Chen, M.W., Kotaka, M., Vonrhein, C., Bricogne, G., Rao, F., Chuah, M.L.C., Svergun, D., Schneider, G., Liang, Z.-X., Lescar, J.&amp;#160; (2012). Signaling. J. Bacteriol. 194: 4837&amp;#8211;4846&lt;/p&gt; &lt;p&gt;&amp;#160;&lt;/p&gt;


Author(s):  
H. Jayalekshmi ◽  
C. Harikrishnan ◽  
Sajin Sali ◽  
N. Kaushik ◽  
Norin Mary G. Victus ◽  
...  

Objective: The present study attempted to evaluate the anti-biofilm activity of D-amino acids (D-AAs) on Pseudomonas aeruginosa and determine if the combination of D-AAs with tetracycline enhances the anti-biofilm activity in vitro and ex vivo.Methods: Different D-AAs were tested for antibiofilm activity against wild type P. aeruginosa PAO1 and two multidrug resistant P. aeruginosa clinical strains in the presence of sub inhibitory concentrations of tetracycline using crystal violet microtitre plate assay. Results were further validated using in vitro wound dressing and ex vivo porcine skin models followed by cytotoxicity and hemocompatibility studies.Results: D-tryptophan (5 mmol) showed 61 % reduction in biofilm formation of P. aeruginosa. Interestingly combinatorial effect of 5 mmol D-tryptophan and 0.5 minimum inhibitory concentration (MIC) (7.5µg/ml) tetracycline showed 90% reduction in biofilm formation. 5 mmol D-methionine shows 28 % reduction and combination with tetracycline shows 41% reduction in biofilm formation of P. aeruginosa. D-leucine and D-tyrosine alone or in combination with tetracycline did not show significant anti-biofilm activity. D tryptophan-tetracycline combination could reduce 80 % and 77 % reduction in biofilm formation in two multi drug resistant P. aeruginosa clinical strains. D-tryptophan-tetracycline-combination could also reduce 76% and 66% reduction in biofilm formation in wound dressing model and porcine skin explant respectively. The cytotoxicity and hemocompatibility studies did not show significant toxicity when this combination was used.Conclusion: The results established the potential therapeutic application of D-tryptophan alone or in combination with tetracycline for treating biofilm associated clinical problems caused by P. aeruginosa.


2019 ◽  
Vol 12 (4) ◽  
pp. 186 ◽  
Author(s):  
Raffaella Campana ◽  
Alessio Merli ◽  
Michele Verboni ◽  
Francesca Biondo ◽  
Gianfranco Favi ◽  
...  

A small library of sugar-based (i.e., glucose, mannose and lactose) monoesters containing hydrophobic aliphatic or aromatic tails were synthesized and tested. The antimicrobial activity of the compounds against a target panel of Gram-positive, Gram-negative and fungi was assessed. Based on this preliminary screening, the antibiofilm activity of the most promising molecules was evaluated at different development times of selected food-borne pathogens (E. coli, L. monocytogenes, S. aureus, S. enteritidis). The antibiofilm activity during biofilm formation resulted in the following: mannose C10 > lactose biphenylacetate > glucose C10 > lactose C10. Among them, mannose C10 and lactose biphenylacetate showed an inhibition for E. coli 97% and 92%, respectively. At MICs values, no toxicity was observed on Caco-2 cell line for all the examined compounds. Overall, based on these results, all the sugar-based monoesters showed an interesting profile as safe antimicrobial agents. In particular, mannose C10 and lactose biphenylacetate are the most promising as possible biocompatible and safe preservatives for pharmaceutical and food applications.


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