N-Nonyloxypentyl-l-Deoxynojirimycin Inhibits Growth, Biofilm Formation and Virulence Factors Expression of Staphylococcus aureus

Staphylococcus aureus is one of the major causes of hospital- and community-associated bacterial infections throughout the world, which are difficult to treat due to the rising number of drug-resistant strains. New molecules displaying potent activity against this bacterium are urgently needed. In this study, d- and l-deoxynojirimycin (DNJ) and a small library of their N-alkyl derivatives were screened against S. aureus ATCC 29213, with the aim to identify novel candidates with inhibitory potential. Among them, N-nonyloxypentyl-l-DNJ (l-NPDNJ) proved to be the most active compound against S. aureus ATCC 29213 and its clinical isolates, with the minimum inhibitory concentration (MIC) value of 128 μg/mL. l-NPDNJ also displayed an additive effect with gentamicin and oxacillin against the gentamicin- and methicillin-resistant S. aureus isolate 00717. Sub-MIC values of l-NPDNJ affected S. aureus biofilm development in a dose-dependent manner, inducing a strong reduction in biofilm biomass. Moreover, real-time reverse transcriptase PCR analysis revealed that l-NPDNJ effectively inhibited at sub-MIC values the transcription of the spa, hla, hlb and sea virulence genes, as well as the agrA and saeR response regulator genes.

Download Full-text

Rosmarinic Acid Interaction with Planktonic and Biofilm Staphylococcus aureus

Natural Product Communications ◽

10.1177/1934578x1300801223 ◽

2013 ◽

Vol 8 (12) ◽

pp. 1934578X1300801 ◽

Cited By ~ 5

Author(s):

Lívia Slobodníková ◽

Silvia Fialová ◽

Helena Hupková ◽

Daniel Grančai

Keyword(s):

Staphylococcus Aureus ◽

Rosmarinic Acid ◽

Antibacterial Activities ◽

Microtiter Plate ◽

Dependent Manner ◽

Early Stages ◽

Microtiter Plates ◽

Sole Agent ◽

The Subject ◽

Biofilm Development

The subject of study was the evaluation of antibacterial activities of rosmarinic acid (RA) on clinical Staphylococcus aureus strains obtained from catheter-related infections. Minimal inhibitory (MIC) and minimal bactericidal concentrations (MBC) of RA were tested by broth microdilution assay. Biofilm-eradication activity was detected on 24-hour biofilm in microtiter plates using a regrowth technique; activity on biofilm formation was measured by a microtiter plate method after RA application to bacterial samples after 0, 1, 3 and 6 hours of biofilm development. RA had antimicrobial activity on all tested strains in concentrations from 625 to 1250 μg.mL−1 (MICs equal to MBCs). No biofilm-eradication activity on 24-hour biofilm was observed in the tested range of concentrations (from 156 to 5000 μg.mL−1). Subinhibitory RA concentrations suppressed the biofilm production, when applied at early stages of its development. Concentrations lower than subinhibitory stimulated the biofilm mass production in a concentration- and time-dependent manner. Considering our results, RA could be a candidate for a topical antimicrobial agent with killing activity on planktonic forms of bacteria and suppressing activity in the early stages of biofilm development, but probably not for the therapy of catheter-related infections as a sole agent.

Download Full-text

CidR and CcpA Synergistically Regulate Staphylococcus aureus cidABC Expression

Journal of Bacteriology ◽

10.1128/jb.00371-19 ◽

2019 ◽

Vol 201 (23) ◽

Cited By ~ 1

Author(s):

Marat R. Sadykov ◽

Ian H. Windham ◽

Todd J. Widhelm ◽

Vijaya Kumar Yajjala ◽

Sean M. Watson ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Carbon Metabolism ◽

Protein A ◽

Transcriptional Regulators ◽

Extracellular Dna ◽

Regulatory Control ◽

Pcr Analysis ◽

Mobility Shift ◽

Content Type ◽

Biofilm Development

ABSTRACT The death and lysis of a subpopulation of Staphylococcus aureus cells during biofilm development benefit the whole bacterial population through the release of an important component of the biofilm matrix, extracellular DNA. Previously, we have demonstrated that these processes are affected by the gene products of the cidABC operon, the expression of which is controlled by the LysR-type transcriptional regulator, CidR. In this study, we characterized cis- and trans-acting elements essential for the induction of the cidABC operon. In addition to a CidR-binding site located within the cidABC promoter region, sequence analysis revealed the presence of a putative catabolite responsive element (cre box), suggestive of the involvement of the catabolite control protein A (CcpA) in the regulation of cidABC expression. This was confirmed using electrophoretic mobility shift assays and real-time reverse transcriptase PCR analysis demonstrating the direct positive control of cidABC transcription by the master regulator of carbon metabolism. Furthermore, the importance of CcpA and the identified cre site for the induction of the cidABC operon was demonstrated by examining the expression of PcidABC-lacZ reporter fusions in various mutant strains in which the genes involved in carbon metabolism and carbon catabolite repression were disrupted. Together the results of this study demonstrate the necessity of both transcriptional regulators, CidR and CcpA, for the induction of the cidABC operon and reveal the complexity of molecular interactions controlling its expression. IMPORTANCE This work focuses on the characterization of cis- and trans-acting elements essential for the induction of the cidABC operon in S. aureus. The results of this study are the first to demonstrate the synergistic control of cidABC expression by transcriptional regulators CidR and CcpA during carbohydrate metabolism. We established that the full induction of cidABC expression depends on the metabolic state of bacteria and requires both CidR and CcpA. Together, these findings delineate regulatory control of cidABC expression under different metabolic conditions and provide important new insights into our understanding of cell death mechanisms during biofilm development in S. aureus.

Download Full-text

Protein A-Mediated Multicellular Behavior in Staphylococcus aureus

Journal of Bacteriology ◽

10.1128/jb.01222-08 ◽

2008 ◽

Vol 191 (3) ◽

pp. 832-843 ◽

Cited By ~ 170

Author(s):

Nekane Merino ◽

Alejandro Toledo-Arana ◽

Marta Vergara-Irigaray ◽

Jaione Valle ◽

Cristina Solano ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Biofilm Formation ◽

Protein A ◽

Surface Proteins ◽

Growth Media ◽

Dependent Manner ◽

Chronic Infections ◽

The Matrix ◽

Implanted Medical Devices ◽

Biofilm Development

ABSTRACT The capacity of Staphylococcus aureus to form biofilms on host tissues and implanted medical devices is one of the major virulence traits underlying persistent and chronic infections. The matrix in which S. aureus cells are encased in a biofilm often consists of the polysaccharide intercellular adhesin (PIA) or poly-N-acetyl glucosamine (PNAG). However, surface proteins capable of promoting biofilm development in the absence of PIA/PNAG exopolysaccharide have been described. Here, we used two-dimensional nano-liquid chromatography and mass spectrometry to investigate the composition of a proteinaceous biofilm matrix and identified protein A (spa) as an essential component of the biofilm; protein A induced bacterial aggregation in liquid medium and biofilm formation under standing and flow conditions. Exogenous addition of synthetic protein A or supernatants containing secreted protein A to growth media induced biofilm development, indicating that protein A can promote biofilm development without being covalently anchored to the cell wall. Protein A-mediated biofilm formation was completely inhibited in a dose-dependent manner by addition of serum, purified immunoglobulin G, or anti-protein A-specific antibodies. A murine model of subcutaneous catheter infection unveiled a significant role for protein A in the development of biofilm-associated infections, as the amount of protein A-deficient bacteria recovered from the catheter was significantly lower than that of wild-type bacteria when both strains were used to coinfect the implanted medical device. Our results suggest a novel role for protein A complementary to its known capacity to interact with multiple immunologically important eukaryotic receptors.

Download Full-text

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.

Download Full-text

ω-Hydroxyemodin Limits Staphylococcus aureus Quorum Sensing-Mediated Pathogenesis and Inflammation

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.04564-14 ◽

2015 ◽

Vol 59 (4) ◽

pp. 2223-2235 ◽

Cited By ~ 63

Author(s):

Seth M. Daly ◽

Bradley O. Elmore ◽

Jeffrey S. Kavanaugh ◽

Kathleen D. Triplett ◽

Mario Figueroa ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Quorum Sensing ◽

Immune Cell ◽

Solid Phase ◽

Response Regulator ◽

Dependent Manner ◽

Accessory Gene ◽

Antibiotic Resistant ◽

Content Type

ABSTRACTAntibiotic-resistant pathogens are a global health threat. Small molecules that inhibit bacterial virulence have been suggested as alternatives or adjuncts to conventional antibiotics, as they may limit pathogenesis and increase bacterial susceptibility to host killing.Staphylococcus aureusis a major cause of invasive skin and soft tissue infections (SSTIs) in both the hospital and community settings, and it is also becoming increasingly antibiotic resistant. Quorum sensing (QS) mediated by the accessory gene regulator (agr) controls virulence factor production essential for causing SSTIs. We recently identified ω-hydroxyemodin (OHM), a polyhydroxyanthraquinone isolated from solid-phase cultures ofPenicillium restrictum, as a suppressor of QS and a compound sought for the further characterization of the mechanism of action. At concentrations that are nontoxic to eukaryotic cells and subinhibitory to bacterial growth, OHM preventedagrsignaling by all fourS. aureus agralleles. OHM inhibited QS by direct binding to AgrA, the response regulator encoded by theagroperon, preventing the interaction of AgrA with theagrP2 promoter. Importantly, OHM was efficacious in a mouse model ofS. aureusSSTI. Decreased dermonecrosis with OHM treatment was associated with enhanced bacterial clearance and reductions in inflammatory cytokine transcription and expression at the site of infection. Furthermore, OHM treatment enhanced the immune cell killing ofS. aureusin vitroin anagr-dependent manner. These data suggest that bacterial disarmament through the suppression ofS. aureusQS may bolster the host innate immune response and limit inflammation.

Download Full-text

Quantitative PCR analysis of genes expressed during biofilm development of methicillin resistant Staphylococcus aureus (MRSA)

Infection Genetics and Evolution ◽

10.1016/j.meegid.2013.05.002 ◽

2013 ◽

Vol 18 ◽

pp. 106-112 ◽

Cited By ~ 46

Author(s):

Salman Sahab Atshan ◽

Mariana Nor Shamsudin ◽

Arunkumar Karunanidhi ◽

Alex van Belkum ◽

Leslie Than Thian Lung ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Quantitative Pcr ◽

Methicillin Resistant Staphylococcus Aureus ◽

Pcr Analysis ◽

Methicillin Resistant ◽

Biofilm Development

Download Full-text

Discovery of Antivirulence Agents against Methicillin-Resistant Staphylococcus aureus

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00269-13 ◽

2013 ◽

Vol 57 (8) ◽

pp. 3645-3652 ◽

Cited By ~ 72

Author(s):

Varandt Khodaverdian ◽

Michelle Pesho ◽

Barbara Truitt ◽

Lucy Bollinger ◽

Parita Patel ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Virtual Screening ◽

Virulence Factors ◽

Small Molecule ◽

Response Regulator ◽

The United States ◽

Dependent Manner ◽

Methicillin Resistant ◽

Content Type ◽

Antivirulence Agents

ABSTRACTAntivirulence agents inhibit the production of disease-causing virulence factors but are neither bacteriostatic nor bactericidal. Antivirulence agents against methicillin-resistantStaphylococcus aureus(MRSA) strain USA300, the most widespread community-associated MRSA strain in the United States, were discovered by virtual screening against the response regulator AgrA, which acts as a transcription factor for the expression of several of the most prominentS. aureustoxins and virulence factors involved in pathogenesis. Virtual screening was followed by similarity searches in the databases of commercial vendors. The small-molecule compounds discovered inhibit the production of the toxins alpha-hemolysin and phenol-soluble modulin α in a dose-dependent manner without inhibiting bacterial growth. These antivirulence agents are small-molecule biaryl compounds in which the aromatic rings either are fused or are separated by a short linker. One of these compounds is the FDA-approved nonsteroidal anti-inflammatory drug diflunisal. This represents a new use for an old drug. Antivirulence agents might be useful in prophylaxis and as adjuvants in antibiotic therapy for MRSA infections.

Download Full-text

5-Dodecanolide interferes with biofilm formation and reduces the virulence of Methicillin-resistant Staphylococcus aureus (MRSA) through up regulation of agr system

Scientific Reports ◽

10.1038/s41598-019-50207-y ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 16

Author(s):

Alaguvel Valliammai ◽

Sivasamy Sethupathy ◽

Arumugam Priya ◽

Anthonymuthu Selvaraj ◽

James Prabhanand Bhaskar ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Biofilm Formation ◽

Bacterial Infections ◽

Pcr Analysis ◽

High Morbidity ◽

Healthy Human ◽

Methicillin Resistant ◽

Gene Expression Studies ◽

Human Infections

Abstract Methicillin resistant Staphylococcus aureus (MRSA) is a predominant human pathogen with high morbidity that is listed in the WHO high priority pathogen list. Being a primary cause of persistent human infections, biofilm forming ability of S. aureus plays a pivotal role in the development of antibiotic resistance. Hence, targeting biofilm is an alternative strategy to fight bacterial infections. The present study for the first time demonstrates the non-antibacterial biofilm inhibitory efficacy of 5-Dodecanolide (DD) against ATCC strain and clinical isolates of S. aureus. In addition, DD is able to inhibit adherence of MRSA on human plasma coated Titanium surface. Further, treatment with DD significantly reduced the eDNA synthesis, autoaggregation, staphyloxanthin biosynthesis and ring biofilm formation. Reduction in staphyloxanthin in turn increased the susceptibility of MRSA to healthy human blood and H2O2 exposure. Quantitative PCR analysis revealed the induced expression of agrA and agrC upon DD treatment. This resulted down regulation of genes involved in biofilm formation such as fnbA and fnbB and up regulation of RNAIII, hld, psmα and genes involved in biofilm matrix degradation such as aur and nuc. Inefficacy of DD on the biofilm formation of agr mutant further validated the agr mediated antibiofilm potential of DD. Notably, DD was efficient in reducing the in vivo colonization of MRSA in Caenorhabditis elegans. Results of gene expression studies and physiological assays unveiled the agr mediated antibiofilm efficacy of DD.

Download Full-text

Broad Spectrum Antimicrobial Activity of Licochalcones A and E against MDR (Multidrug Resistant) Strains of Clinical Origin

Natural Product Communications ◽

10.1177/1934578x1701201123 ◽

2017 ◽

Vol 12 (11) ◽

pp. 1934578X1701201 ◽

Cited By ~ 1

Author(s):

Jung-Eun Kim ◽

Goo Yoon ◽

Jung-Hyun Shim ◽

Seung-Sik Cho

Keyword(s):

Staphylococcus Aureus ◽

Antimicrobial Activity ◽

Bacterial Infections ◽

Multidrug Resistant ◽

Drug Resistant ◽

Baseline Information ◽

Resistant Strains ◽

Glycyrrhiza Inflata ◽

Drug Resistant Strains ◽

Potential Use

The aim of this study was to evaluate the antibacterial activity of the licochalcones A (1) and E (2) against drug resistant strains of clinical origin. The results indicate that the licochalcones had a broad inhibitory activity against tested bacteria. Compared to vancomycin and teicoplanin, these compounds provided weaker activity against non-MDR Staphylococcus aureus and Enterococcus but broader activity against MRSA and VRE strains. The results provide promising baseline information for the potential use of 1 and 2 from Glycyrrhiza inflata in the treatment of drug resistant bacterial infections.

Download Full-text

Mode of action of Elasnin as biofilm-formation eradicator of Methicillin-Resistant Staphylococcus aureus

10.21203/rs.3.rs-752510/v1 ◽

2021 ◽

Author(s):

Lexin Long ◽

Jordy Evan Sulaiman ◽

Yao Xiao ◽

Aifang Cheng ◽

Ruojun WANG ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Biofilm Formation ◽

Bacterial Infections ◽

Penicillin G ◽

Dependent Manner ◽

Resistance Development ◽

Methicillin Resistant ◽

Wide Range ◽

Increased Sensitivity ◽

Low Cytotoxicity

Abstract Biofilm is made by microbes and often offers protection by making them more tolerant, resistant, and resilient to wide-range antimicrobials. Biofilm-related infections account for more than 80% of human bacterial infections and are especially prevalent in chronic tissue and device-related infections. Owing to the great challenge in treating biofilms, novel and effective antibiofilm compounds urgently need to be identified. We herein identified elasnin as a potent biofilm-targeting compound against methicillin-resistant Staphylococcus aureus (MRSA) through bioassay-guided isolation of bioactive compounds from Actinobacteria. Elasnin effectively inhibited biofilm formation and eradicated pre-formed biofilms of MRSA, and it displays low cytotoxicity with a low risk of resistance development. Transcriptomic and proteomic analyses combined with confocal microscopy observation revealed that elasnin destroyed the biofilm matrix in a time-dependent manner and interfered with cell cycle during the exponential phase, primarily by repressing the expression of virulence factors. Moreover, the biofilm cells released from elasnin treatment showed increased sensitivity to penicillin G. Overall, this study identified elasnin as a promising biofilm-eradicating compound against MRSA and shed light on its action mechanism.

Download Full-text