Enhancement of Antibiofilm Activity of Ciprofloxacin against Staphylococcus aureus by Administration of Antimicrobial Peptides

Staphylococcus aureus can develop resistance by mutation, transfection or biofilm formation. Resistance was induced in S. aureus by growth in sub-inhibitory concentrations of ciprofloxacin for 30 days. The ability of the antimicrobials to disrupt biofilms was determined using crystal violet and live/dead staining. Effects on the cell membranes of biofilm cells were evaluated by measuring release of dyes and ATP, and nucleic acids. None of the strains developed resistance to AMPs while only S. aureus ATCC 25923 developed resistance (128 times) to ciprofloxacin after 30 passages. Only peptides reduced biofilms of ciprofloxacin-resistant cells. The antibiofilm effect of melimine with ciprofloxacin was more (27%) than with melimine alone at 1X MIC (p < 0.001). Similarly, at 1X MIC the combination of Mel4 and ciprofloxacin produced more (48%) biofilm disruption than Mel4 alone (p < 0.001). Combinations of either of the peptides with ciprofloxacin at 2X MIC released ≥ 66 nM ATP, more than either peptide alone (p ≤ 0.005). At 2X MIC, only melimine in combination with ciprofloxacin released DNA/RNA which was three times more than that released by melimine alone (p = 0.043). These results suggest the potential use of melimine and Mel4 with conventional antibiotics for the treatment of S. aureus biofilms.

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Enhancement of Antibiofilm Activity of Ciprofloxacin against Staphylococcus aureus by Administration of Antimicrobial Peptides

10.20944/preprints202108.0451.v1 ◽

2021 ◽

Author(s):

Muhammad Yasir ◽

Debarun Dutta ◽

Mark Duncan Perry Willcox

Keyword(s):

Staphylococcus Aureus ◽

Antimicrobial Peptides ◽

Nucleic Acids ◽

Crystal Violet ◽

Biofilm Formation ◽

Cell Membranes ◽

Antibiofilm Activity ◽

Biofilm Disruption ◽

Conventional Antibiotics ◽

Potential Use

Staphylococcus aureus can develop resistance by mutation, tranfection or biofilm formation. Resistance was induced in S. aureus by growth in sub-inhibitory concentrations of ciprofloxacin for 30 days. The ability of the antimicrobials to disrupt biofilms was determined using crystal violet and live/dead staining. Effects on the cell membranes of biofilm cells was evaluated by measuring release of dyes and ATP and nucleic acids. S. aureus did not develop resistance to the AMPs but resistance increased to ciprofloxacin by 128 times after 30 passages. Only peptides reduced biofilms of ciprofloxacin resistant cells. The antibiofilm effect of melimine with ciprofloxacin was more (27%) than with melimine alone at 1X MIC (p < 0.001). Similarly, at 1X MIC the combination of Mel4 and ciprofloxacin produced more (48%) biofilm disruption than Mel4 alone (p < 0.001). Combinations of either of the peptides with ciprofloxacin at 2X MIC released  66 nM ATP, more than either peptide alone (p  0.005). At 2X MIC, only melimine in combination with ciprofloxacin released DNA/RNA which was 3 times more than released by melimine alone (p = 0.043). These results suggest the potential use of melimine and Mel4 with conventional antibiotics for the treatments of S. aureus biofilms.

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Inhibition of Biofilm Formation by Esomeprazole in Pseudomonas aeruginosa and Staphylococcus aureus

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00544-12 ◽

2012 ◽

Vol 56 (8) ◽

pp. 4360-4364 ◽

Cited By ~ 17

Author(s):

Vandana Singh ◽

Vaneet Arora ◽

M. Jahangir Alam ◽

Kevin W. Garey

Keyword(s):

Staphylococcus Aureus ◽

Pseudomonas Aeruginosa ◽

Bacterial Growth ◽

Biofilm Formation ◽

Antimicrobial Properties ◽

Antibiofilm Activity ◽

Biofilm Growth ◽

Content Type ◽

Conventional Antibiotics ◽

And Staphylococcus Aureus

ABSTRACTStaphylococcus aureusandPseudomonas aeruginosaare common nosocomial pathogens responsible for biofilm-associated infections. Proton pump inhibitors (PPI), such as esomeprazole, may have novel antimicrobial properties. The objective of this study was to assess whether esomeprazole prevents sessile bacterial growth and biofilm formation and whether it may have synergistic killing effects with standard antibiotics. The antibiofilm activity of esomeprazole at 0.25 mM was tested against two strains each ofS. aureusandP. aeruginosa. Bacterial biofilms were prepared using a commercially available 96-peg-plate Calgary biofilm device. Sessile bacterial CFU counts and biomass were assessed during 72 hours of esomeprazole exposure. The killing activities after an additional 24 hours of vancomycin (againstS. aureus) and meropenem (againstP. aeruginosa) treatment with or without preexposure to esomeprazole were also assessed by CFU and biomass analyses.P. aeruginosaandS. aureusstrains exposed to esomeprazole displayed decreased sessile bacterial growth and biomass (P< 0.001, each parameter). After 72 h of exposure, there was a 1-log10decrease in the CFU/ml of esomeprazole-exposedP. aeruginosaandS. aureusstrains compared to controls (P< 0.001). After 72 h of exposure, measured absorbance was 100% greater inP. aeruginosacontrol strains than in esomeprazole-exposed strains (P< 0.001). Increased killing and decreased biomass were observed for esomeprazole-treated bacteria compared to untreated controls exposed to conventional antibiotics (P< 0.001, each parameter). Reduced biofilm growth after 24 h was visibly apparent by light micrographs forP. aeruginosaandS. aureusisolates exposed to esomeprazole compared to untreated controls. In conclusion, esomeprazole demonstrated an antibiofilm effect against biofilm-producingS. aureusandP. aeruginosa.

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Antimicrobial and Antibiofilm Peptides

Biomolecules ◽

10.3390/biom10040652 ◽

2020 ◽

Vol 10 (4) ◽

pp. 652 ◽

Cited By ~ 12

Author(s):

Angela Di Somma ◽

Antonio Moretta ◽

Carolina Canè ◽

Arianna Cirillo ◽

Angela Duilio

Keyword(s):

Antimicrobial Peptides ◽

Biofilm Formation ◽

Bacterial Resistance ◽

Antimicrobial Agents ◽

Multidrug Resistant ◽

Resistant Bacteria ◽

Antibiofilm Activity ◽

Chronic Infections ◽

Planktonic Bacteria ◽

Hostile Environments

The increasing onset of multidrug-resistant bacteria has propelled microbiology research towards antimicrobial peptides as new possible antibiotics from natural sources. Antimicrobial peptides are short peptides endowed with a broad range of activity against both Gram-positive and Gram-negative bacteria and are less prone to trigger resistance. Besides their activity against planktonic bacteria, many antimicrobial peptides also show antibiofilm activity. Biofilms are ubiquitous in nature, having the ability to adhere to virtually any surface, either biotic or abiotic, including medical devices, causing chronic infections that are difficult to eradicate. The biofilm matrix protects bacteria from hostile environments, thus contributing to the bacterial resistance to antimicrobial agents. Biofilms are very difficult to treat, with options restricted to the use of large doses of antibiotics or the removal of the infected device. Antimicrobial peptides could represent good candidates to develop new antibiofilm drugs as they can act at different stages of biofilm formation, on disparate molecular targets and with various mechanisms of action. These include inhibition of biofilm formation and adhesion, downregulation of quorum sensing factors, and disruption of the pre-formed biofilm. This review focuses on the proprieties of antimicrobial and antibiofilm peptides, with a particular emphasis on their mechanism of action, reporting several examples of peptides that over time have been shown to have activity against biofilm.

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Antimicrobial and Antibiofilm Activity against Staphylococcus aureus of Opuntia ficus-indica (L.) Mill. Cladode Polyphenolic Extracts

Antioxidants ◽

10.3390/antiox8050117 ◽

2019 ◽

Vol 8 (5) ◽

pp. 117 ◽

Cited By ~ 17

Author(s):

Federica Blando ◽

Rossella Russo ◽

Carmine Negro ◽

Luigi De Bellis ◽

Stefania Frassinetti

Keyword(s):

Staphylococcus Aureus ◽

Antioxidant Activity ◽

Antioxidant Capacity ◽

Biofilm Formation ◽

Ex Vivo ◽

Trolox Equivalent Antioxidant Capacity ◽

Total Phenolic ◽

Antibiofilm Activity ◽

Opuntia Ficus Indica

Plant extracts are a rich source of natural compounds with antimicrobial properties, which are able to prevent, at some extent, the growth of foodborne pathogens. The aim of this study was to investigate the potential of polyphenolic extracts from cladodes of Opuntia ficus-indica (L.) Mill. to inhibit the growth of some enterobacteria and the biofilm formation by Staphylococcus aureus. Opuntia ficus-indica cladodes at two stages of development were analysed for total phenolic content and antioxidant activity by Oxygen Radical Absorbance Capacity (ORAC) and Trolox equivalent antioxidant capacity (TEAC) (in vitro assays) and by cellular antioxidant activity in red blood cells (CAA-RBC) (ex vivo assay). The Liquid Chromatography Time-of-Flight Mass Spectrometry (LC/MS–TOF) analysis of the polyphenolic extracts revealed high levels of piscidic acid, eucomic acid, isorhamnetin derivatives and rutin, particularly in the immature cladode extracts. Opuntia cladodes extracts showed a remarkable antioxidant activity (in vitro and ex vivo), a selective inhibition of the growth of Gram-positive bacteria, and an inhibition of Staphylococcus aureus biofilm formation. Our results suggest and confirm that Opuntia ficus-indica cladode extracts could be employed as functional food, due to the high polyphenolic content and antioxidant capacity, and used as natural additive for food process control and food safety.

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Antibacterial and Antibiofilm Potential of Sea Anemone (Stichodactyla haddoni)-Isolated Vibrio (V. parahaemolyticus and V. alginolyticus), and Pseudoalteromonas (P. gelatinilytica and P. piscicida) Against Staphylococcus aureus and Pseudomonas aeruginosa

Journal of Kermanshah University of Medical Sciences ◽

10.5812/jkums.108653 ◽

2021 ◽

Vol 25 (1) ◽

Author(s):

Neda Fazeli ◽

Akram Sadat Naeemi ◽

Seyed Amir Hossein Jalali ◽

Hojjatollah Zamani

Keyword(s):

Staphylococcus Aureus ◽

Pseudomonas Aeruginosa ◽

Biofilm Formation ◽

Sea Anemone ◽

Broth Culture ◽

Antibiofilm Activity ◽

Microtiter Plates ◽

Protease Enzyme ◽

Molecular Identification Methods ◽

Stichodactyla Haddoni

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.

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Piperine Exhibits Promising Antibiofilm Activity Against Staphylococcus Aureus by Accumulating Reactive Oxygen Species (ROS)

10.21203/rs.3.rs-514513/v1 ◽

2021 ◽

Author(s):

Sharmistha Das ◽

Payel Paul ◽

Sudipta Chatterjee ◽

Poulomi Chakraborty ◽

Ranojit K. Sarker ◽

...

Keyword(s):

Reactive Oxygen Species ◽

Staphylococcus Aureus ◽

Biofilm Formation ◽

Reactive Oxygen ◽

Antibiofilm Activity ◽

Host Immune System ◽

Oxygen Species ◽

Bacterial Cells ◽

Protein Assay ◽

Area Of Concern

Abstract Biofilm, an aggregated form of microbial existence has been a major area of concern in the healthcare units. These sessile microbes not only protect themselves from the host immune system but also exhibit high resistance against several antimicrobials. One such widely reported Gram-positive pathogen is Staphylococcus aureus. This human commensal is known to cause severe harmful diseases like bacteremia, sepsis, pneumonia, etc. Thus, strategies need to be undertaken to deal with such biofilm challenges. In this respect, we aimed to inhibit microbial biofilm formation of Staphylococcus aureus under the influence of a natural compound, piperine. Our study revealed that the higher concentrations of piperine exhibited considerable antimicrobial activity against Staphylococcus aureus. Hence, lower concentrations of piperine were tested to examine its antibiofilm activity. Several experiments like crystal violet (CV) assay, total biofilm protein assay, and fluorescence microscopy observation established that lower concentrations (8 µg/mL and 16 µg/mL) of piperine showed efficient antibiofilm activity against Staphylococcus aureus. It was also noticed that the lower concentrations of piperine did not compromise the microbial growth of Staphylococcus aureus while exhibiting antibiofilm activity. In this connection, we also noticed that the lower concentrations of piperine showed a considerable reduction in microbial metabolic activity. Furthermore, we observed that the compound was found to accumulate reactive oxygen species in the bacterial cells that could play an important role in the inhibition of biofilm formation. Thus, piperine could be considered as a potential antibiofilm agent against the biofilm formation caused by Staphylococcus aureus.

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Tannic Acid Inhibits Staphylococcus aureus Surface Colonization in an IsaA-Dependent Manner

Infection and Immunity ◽

10.1128/iai.00877-12 ◽

2012 ◽

Vol 81 (2) ◽

pp. 496-504 ◽

Cited By ~ 59

Author(s):

David E. Payne ◽

Nicholas R. Martin ◽

Katherine R. Parzych ◽

Alex H. Rickard ◽

Adam Underwood ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Tannic Acid ◽

Biofilm Formation ◽

Antibiofilm Activity ◽

Host Immune System ◽

Dependent Manner ◽

Content Type ◽

Surface Colonization ◽

Biofilm Development ◽

Insight Into

ABSTRACTStaphylococcus aureusis a human commensal and pathogen that is capable of forming biofilms on a variety of host tissues and implanted medical devices. Biofilm-associated infections resist antimicrobial chemotherapy and attack from the host immune system, making these infections particularly difficult to treat. In order to gain insight into environmental conditions that influenceS. aureusbiofilm development, we screened a library of small molecules for the ability to inhibitS. aureusbiofilm formation. This led to the finding that the polyphenolic compound tannic acid inhibitsS. aureusbiofilm formation in multiple biofilm models without inhibiting bacterial growth. We present evidence that tannic acid inhibitsS. aureusbiofilm formation via a mechanism dependent upon the putative transglycosylase IsaA. Tannic acid did not inhibit biofilm formation of anisaAmutant. Overexpression of wild-type IsaA inhibited biofilm formation, whereas overexpression of a catalytically dead IsaA had no effect. Tannin-containing drinks like tea have been found to reduce methicillin-resistantS. aureusnasal colonization. We found that black tea inhibitedS. aureusbiofilm development and that anisaAmutant resisted this inhibition. Antibiofilm activity was eliminated from tea when milk was added to precipitate the tannic acid. Finally, we developed a rodent model forS. aureusthroat colonization and found that tea consumption reducedS. aureusthroat colonization via anisaA-dependent mechanism. These findings provide insight into a molecular mechanism by which commonly consumed polyphenolic compounds, such as tannins, influenceS. aureussurface colonization.

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Citral modulates virulence factors in methicillin-resistant Staphylococcus aureus

Scientific Reports ◽

10.1038/s41598-021-95971-y ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Hellen Braga Martins Oliveira ◽

Nathan das Neves Selis ◽

Beatriz Almeida Sampaio ◽

Manoel Neres Santos Júnior ◽

Suzi Pacheco de Carvalho ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Antimicrobial Activity ◽

Biofilm Formation ◽

Production Capacity ◽

High Morbidity ◽

Methicillin Resistant ◽

Biofilm Production ◽

Initial Stage ◽

Mrsa Strains ◽

Conventional Antibiotics

AbstractMethicillin-resistant Staphylococcus aureus (MRSA) is responsible for high morbidity and mortality rates. Citral has been studied in the pharmaceutical industry and has shown antimicrobial activity. This study aimed to analyze the antimicrobial activity of citral in inhibiting biofilm formation and modulating virulence genes, with the ultimate goal of finding a strategy for treating infections caused by MRSA strains. Citral showed antimicrobial activity against MRSA isolates with minimum inhibitory concentration (MIC) values between 5 mg/mL (0.5%) and 40 mg/mL (4%), and minimum bactericidal concentration (MBC) values between 10 mg/mL (1%) and 40 mg/mL (4%). The sub-inhibitory dose was 2.5 mg/mL (0.25%). Citral, in an antibiogram, modulated synergistically, antagonistically, or indifferent to the different antibiotics tested. Prior to evaluating the antibiofilm effects of citral, we classified the bacteria according to their biofilm production capacity. Citral showed greater efficacy in the initial stage, and there was a significant reduction in biofilm formation compared to the mature biofilm. qPCR was used to assess the modulation of virulence factor genes, and icaA underexpression was observed in isolates 20 and 48. For icaD, seg, and sei, an increase was observed in the expression of ATCC 33,591. No significant differences were found for eta and etb. Citral could be used as a supplement to conventional antibiotics for MRSA infections.

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Lipidation of Temporin-1CEb Derivatives as a Tool for Activity Improvement, Pros and Cons of the Approach

International Journal of Molecular Sciences ◽

10.3390/ijms22136679 ◽

2021 ◽

Vol 22 (13) ◽

pp. 6679

Author(s):

Paulina Kosikowska-Adamus ◽

Emilia Sikorska ◽

Dariusz Wyrzykowski ◽

Aleksandra Walewska ◽

Anna Golda ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Antimicrobial Peptides ◽

Antimicrobial Properties ◽

Carbon Chain ◽

Microbial Pathogens ◽

Multi Drug Resistance ◽

Aggregation State ◽

Conventional Antibiotics ◽

Biofilm Development

The alarming raise of multi-drug resistance among human microbial pathogens makes the development of novel therapeutics a priority task. In contrast to conventional antibiotics, antimicrobial peptides (AMPs), besides evoking a broad spectrum of activity against microorganisms, could offer additional benefits, such as the ability to neutralize toxins, modulate inflammatory response, eradicate bacterial and fungal biofilms or prevent their development. The latter properties are of special interest, as most antibiotics available on the market have limited ability to diffuse through rigid structures of biofilms. Lipidation of AMPs is considered as an effective approach for enhancement of their antimicrobial potential and in vivo stability; however, it could also have undesired impact on selectivity, solubility or the aggregation state of the modified peptides. In the present work, we describe the results of structural modifications of compounds designed based on cationic antimicrobial peptides DK5 and CAR-PEG-DK5, derivatized at their N-terminal part with fatty acids with different lengths of carbon chain. The proposed modifications substantially improved antimicrobial properties of the final compounds and their effectiveness in inhibition of biofilm development as well as eradication of pre-formed 24 h old biofilms of Candida albicans and Staphylococcus aureus. The most active compounds (C5-DK5, C12-DK5 and C12-CAR-PEG-DK5) were also potent against multi-drug resistant Staphylococcus aureus USA300 strain and clinical isolates of Pseudomonas aeruginosa. Both experimental and in silico methods revealed strong correlation between the length of fatty acid attached to the peptides and their final membranolytic properties, tendency to self-assemble and cytotoxicity.

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Small-Molecule Compound SYG-180-2-2 to Effectively Prevent the Biofilm Formation of Methicillin-Resistant Staphylococcus aureus

Frontiers in Microbiology ◽

10.3389/fmicb.2021.770657 ◽

2022 ◽

Vol 12 ◽

Author(s):

Lulin Rao ◽

Yaoguang Sheng ◽

Jiao Zhang ◽

Yanlei Xu ◽

Jingyi Yu ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Small Molecule ◽

Biofilm Formation ◽

Bacterial Biofilm ◽

Alveolar Epithelial Cells ◽

Laser Scanning Microscopy ◽

Confocal Laser ◽

Antibiofilm Activity ◽

Methicillin Resistant ◽

Small Molecule Compound

The resistance of methicillin-resistant Staphylococcus aureus (MRSA) has augmented due to the abuse of antibiotics, bringing about difficulties in the treatment of infection especially with the formation of biofilm. Thus, it is essential to develop antimicrobials. Here we synthesized a novel small-molecule compound, which we termed SYG-180-2-2 (C21H16N2OSe), that had antibiofilm activity. The aim of this study was to demonstrate the antibiofilm effect of SYG-180-2-2 against clinical MRSA isolates at a subinhibitory concentration (4 μg/ml). In this study, it was showed that significant suppression in biofilm formation occurred with SYG-180-2-2 treatment, the inhibition ranged between 65.0 and 85.2%. Subsequently, confocal laser scanning microscopy and a bacterial biofilm metabolism activity assay further demonstrated that SYG-180-2-2 could suppress biofilm. Additionally, SYG-180-2-2 reduced bacterial adhesion and polysaccharide intercellular adhesin (PIA) production. It was found that the expression of icaA and other biofilm-related genes were downregulated as evaluated by RT-qPCR. At the same time, icaR and codY were upregulated when biofilms were treated with SYG-180-2-2. Based on the above results, we speculate that SYG-180-2-2 inhibits the formation of biofilm by affecting cell adhesion and the expression of genes related to PIA production. Above all, SYG-180-2-2 had no toxic effects on human normal alveolar epithelial cells BEAS-2B. Collectively, the small-molecule compound SYG-180-2-2 is a safe and effective antibacterial agent for inhibiting MRSA biofilm.

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