Preexisting Biofilms Research Articles

A perspective on nanomaterials against Campylobacter jejuni biofilm - new control strategies.

Campylobacter jejuni - a Gram-negative bacterium - is considered the fourth cause of diarrheic diseases that can form biofilms (mono and multi-species) or colonize pre-existing biofilms adhering to both, inert or biotic surfaces; its biofilms contribute to transmission through the food chain and survival under harsh environmental conditions. Thus, developing alternatives against this pathogen is compulsory. Nanomaterials have revolutionized the way of fighting infections related to biofilms due to their unique properties compared to traditional antibiotics. Nanomaterials have also been used against C. jejuni based on zinc, titanium, silver, molybdenum, magnesium, cobalt, erbium, lithium, nickel, hydroxide, polyethylene, graphene, lipids, chitosan, and poly(lactic-co-glycolic acid) (PLGA). Those organic and inorganic materials have synthesized nanoparticles, nanofillers, nanowires, nanoferrites, double layers, nanocomposites, and films that have encapsulated, entrapped, coated or doped molecules. Additionally, bare metal nanoparticles have been tested by their antimicrobial activity on planktonic and sessile forms. Therefore, the present review aimed to describe general biology virulence factors, host-pathogen relationships and biofilm formation, as well as nanomaterials and nanoparticles fighting against C. jejuni biofilms. Considerations are presented and placed in perspective.

Pillar[5]arene stabilized gold nanoparticles for the enhanced light-triggered nitric oxide release with antibacterial and antibiofilm activities

The reactivity of guests confined in well-defined host cavities act significantly different from that in bulk systems. In this work, pillar[5]arene not only works as a stabilizer for the fabrication of gold nanoparticles, but also act as an accelerator for the enhanced photo-triggered release of gaseous nitric oxide (NO) from encapsulated NO donor. The host–guest complex can dramatically improve the photoactivity of loaded guest molecule to generate antibacterial NO molecules through the confinement effect of pillar[5]arene cavity. The synergistic photo-triggered localized NO release and photo-thermal effect of host–guest complex demonstrated impressive antibacterial behaviors against Gram-negative bacterial strain Escherichia coli, Gram-positive bacterial strain Staphylococcus aureus and methicillin-resistant Staphylococcus aureus. Moreover, this biocompatible photo-responsive complex can also inhibit bacterial biofilm formation and disrupt the pre-existing biofilms.

Enhancing Antibacterial Properties of Titanium Implants through Covalent Conjugation of Self-Assembling Fmoc-Phe-Phe Dipeptide on Titania Nanotubes.

Bacterial infections and biofilm formation are significant challenges for medical implants. While titanium nanotube engineering improves biocompatibility, it cannot prevent bacterial adhesion and biofilm formation. Optimizing the biomaterial's surface chemistry is vital for its desired functioning in the biological environment. This study demonstrates the covalent conjugating of the self-assembling dipeptide N-fluorenylmethyloxycarbonyl-diphenylalanine (Fmoc-FF) onto titanium nanotube surfaces (TiNTs) without altering the topography. Fmoc-FF peptides, in conjugation with TiNTs, can inhibit biofilm formation, eradicate pre-existing biofilms, and kill bacteria. This functionalization imparts antibacterial properties to the surface while retaining beneficial nanotube topography, synergistically enhancing bioactivity. Surface characterization by XPS, FT-IR, EDS, and SEM confirmed the successful functionalization. Bacterial adhesion experiments showed a significantly improved antibacterial activity of the functionalized TiNT surfaces. This study opens future possibilities for associating biomedical applications such as cell-cell interactions, tissue engineering, and controlled drug delivery of multifunctional self-assembling short peptides with implant materials through surface functionalization.

Antibiofilm and antivirulence efficacy of Pleurotus platypus methanolic extract against Staphylococcus aureus through ROS generation and cell membrane disruption

Multi-drug resistance is recognized as a significant worldwide public health concern in the current century. Biofilm formation further exacerbates bacterial resistance to antibacterial medications, host immunological responses, and phagocytosis, resulting in long-lasting chronic illnesses. Investigating natural resources is a very potent approach for developing alternative anti-infective medications to effectively control multi-drug resistant bacterial infections. In this study, a unique mushroom species namely Pleurotus platypus had been discovered from the Terai–Duars region of West Bengal, India. The myco-chemical profiling and preliminary chemical analysis of Pleurotus platypus methanolic extract determined the significant presence of metabolites belonging to several major chemical classes such as flavonoid, alkaloid, triterpenoid, polyphenol, benzoic acids, coumarin, flavone etc. Most intriguingly, the extract possessed effective antibacterial, antibiofilm and antivirulence properties against Staphylococcus aureus and Methicillin resistant Staphylococcus aureus, one of the most notable drug-resistant opportunistic and nosocomial pathogens. Mechanistically, the mushroom extract enhanced the production of Reactive Oxygen Species (ROS) inside the targeted bacteria, causing alterations in membrane potential, damage to the cellular membrane and further release of intracellular DNA, destined to cell death. Moreover, the methanolic extract reported the eradication of pre-existing biofilms from the urinary catheter surface, hinting towards its future application in the related field. To summarize, Pleurotus platypus methanolic extract could be an excellent alternative antibacterial and antibiofilm therapeutic candidate for the effective management of Staphylococcus infections with improved outcome.

The Influence of Silver-Containing Bionanomaterials Based on Humic Ligands on Biofilm Formation in Opportunistic Pathogens.

The uncontrolled use of antibiotics has led to a global problem of antimicrobial resistance. One of the main mechanisms of bacterial resistance is the formation of biofilms. In order to prevent the growth of antimicrobial resistance, it is crucial to develop new antibacterial agents that are capable of inhibiting the formation of biofilms. This makes this area of research highly relevant today. Promising candidates for these antibacterial agents are new bionanomaterials made from natural humic substances and silver nanoparticles. These substances have the potential to not only directly kill microorganisms but also penetrate biofilms and inhibit their formation. The goal of this study is to synthesize active pharmaceutical substances in the form of bionanomaterials, using ultradispersed silver nanoparticles in a matrix of coal humic substances, perform their characterization (NMR spectroscopy, TEM, and ICP-AES methods), and research their influence on biofilm formation in the most dangerous opportunistic pathogens (E. coli, Methicillin-resistant St. Aureus, K. pneumoniae, P. aeruginosa, St. aureus, A. baumannii, and K. Pneumonia). The results showed that all of the studied bionanomaterials had antibacterial activity against all of the opportunistic pathogens. Furthermore, they were found to have a suppressive effect on both pre-existing biofilms of these bacteria and their formation.

Bioemulsifier from sponge-associated bacteria reduces staphylococcal biofilm

Biofilm formation is a major health concern and studies have been pursued to find compounds able to prevent biofilm establishment and remove pre-existing biofilms. While biosurfactants (BS) have been well-known for possessing antibiofilm activities, bioemulsifiers (BE) are still scarcely explored for this purpose. The present study aimed to evaluate the bioemulsifying properties of cell-free supernatants produced by Bacillaceae and Vibrio strains isolated from marine sponges and investigate their antiadhesive and antibiofilm activities against different pathogenic Gram-positive and Gram-negative bacteria. The BE production by the marine strains was confirmed by the emulsion test, drop-collapsing, oil-displacement, cell hydrophobicity and hemolysis assays. Notably, Bacillus cereus 64BHI1101 displayed remarkable emulsifying activity and the ultrastructure analysis of its BE extract (BE64-1) revealed the presence of structures typically observed in macromolecules composed of polysaccharides and proteins. BE64-1 showed notable antiadhesive and antibiofilm activities against Staphylococcus aureus, with a reduction of adherence of up to 100 % and a dispersion of biofilm of 80 %, without affecting its growth. BE64-1 also showed inhibition of Staphylococcus epidermidis and Escherichia coli biofilm formation and adhesion. Thus, this study provides a starting point for exploring the antiadhesive and antibiofilm activities of BE from sponge-associated bacteria, which could serve as a valuable tool for future research to combat S. aureus biofilms.

Antibacterial and Antibiofilm Effects of L-Carnitine-Fumarate on Oral Streptococcal Strains Streptococcus mutans and Streptococcus sobrinus.

Streptococcus mutans is a major pathogenic habitant of oral caries. Owing to its physiological and biochemical features, it prevails in the form of plaque biofilm together with another important mutans streptococci species, Streptococcus sobrinus. Both species are considered as initiators of cavity lesions, and biofilm is essential to the dental caries process. Compared with the planktonic populations, the biofilm form has higher resistance to environmental conditions and antibiotics. Dental plaques also secure the long-term survival of microorganisms and protection from any stress conditions. To address the need for new antibiofilm agents, we have focused on L-carnitine-fumarate, a fumarate-conjugated quaternary ammonium compound. Using the macro-broth susceptibility testing method, we established its MIC value as 6.0 mg/mL. The MBC value, determined from the broth dilution minimum inhibitory concentration test by sub-culturing it to BHI agar plates, was established as 7.0 mg/mL. Antibiofilm efficacy was tested in 96-well plates coated with saliva using BHI broth supplemented with 1% sucrose as a standard approach. The obtained results allowed us to assess the MIBC as 7.5 mg/mL and the MBBC value as 10.0 mg/mL. The latter concentration also caused approximately 20% eradication of pre-existing biofilm. EPS-rich matrix, forming the core of the biofilm and enabling a confined acidic microenvironment, was also examined and confirmed the effectiveness of 10.0 mg/mL L-carnitine-fumarate concentration in inhibiting EPS formation. Furthermore, the anti-adherent and anti-aciduric impacts of L-carnitine-fumarate were investigated and revealed significant inhibitory effects at sub-MIC concentrations. The influence of L-carnitine-fumarate on the phosphotransferase system was investigated as well. Our results provide a new insight into the antibacterial potential of L-carnitine-fumarate as a valuable compound to be considered for alternative or adjunct anti-caries and antibiofilm preventive approaches.

Isolation and identification of antimicrobial multicyclic terpenoids from the medicinal plant Salvia officinalis and development of a formulation against clinical Staphylococcus aureus strains.

Staphylococcus aureus, particularly multi-drug resistant strains, presents significant challenges in dairy farming due to its role in causing bovine mastitis, which leads to substantial economic losses and limited treatment options. Seeking alternative therapies, we investigated the potential of a topical formulation derived from the medicinal herb Salvia officinalis to combat S. aureus growth and biofilms associated with bovine mastitis. Through systematic extraction in different solvents and fractionation by column chromatography, we isolated and identified three key multicyclic terpenoids-ferruginol, sugiol, and sclareol-exhibiting significant antimicrobial activity. The formulation effectively inhibited biofilm formation, with minimum inhibitory concentration (MIC) values ranging from 0.09 to 0.74mg ml-1 against clinical S. aureus strains, comparable to or lower than those of the pure compounds. Moreover, it displayed robust anti-adhesive properties, reducing biofilm formation by 20%-79% at subinhibitory concentrations. Furthermore, the formulation successfully disrupted pre-existing biofilms, achieving reductions ranging from 30% to 82%. Cytotoxicity assays confirmed the safety of the formulation on mammary epithelial cells, with cell viability maintained at 100% at MIC. Our findings underscore the therapeutic potential of Sa. officinalis-derived compounds in managing bovine mastitis caused by S. aureus, emphasizing their antimicrobial efficacy and safety profile.

Bioinspired ferromagnetic NiFe2O4 nanoparticles: Eradication of fungal and drug-resistant bacterial pathogens and their established biofilm

Nickel ferrite nanoparticles (NiFe2O4 NPs) were synthesized using the medicinally important plant Aloe vera leaf extract, and their structural, morphological, and magnetic properties were characterized by x-ray diffraction (XRD), fourier transform infrared (FTIR), scanning electron microscopy (SEM), energy dispersive x-ray (EDX), and vibrating sample magnetometer (VSM). The synthesized NPs were soft ferromagnetic and spinel in nature, with an average particle size of 22.2 nm. To the best of our understanding, this is the first comprehensive investigation into the antibacterial, anticandidal, antibiofilm, and antihyphal properties of NiFe2O4 NPs against C. albicans as well as drug-resistant gram-positive methicillin-resistant Staphylococcus aureus (MRSA) and gram-negative multidrug resistant Pseudomonas aeruginosa (MDR-P. aeruginosa) bacteria. NiFe2O4 NPs showed potent antimicrobial activity (MIC 1.6–2 mg/mL) against the test pathogens. NiFe2O4 NPs at 0.5 mg/mL suppressed biofilm formation by 49.5–53.1 % in test pathogens. The study found that the NPs not only prevent the formation of biofilm, but also eliminate existing mature biofilms by 50.5–75.79 % at 0.5 mg/mL, which was further validated by SEM. SEM examination revealed a reduction in the number of cells that form biofilms and adhere to the surface. Additionally, it considerably impeded the colonization and aggregation of the biofilm strains on the glass surface. Light microscopic examination demonstrated that NPs effectively prevent the expansion of hyphae, filaments, and yeast-to-hyphae transformation in C. albicans, resulting in a substantial decrease in their ability to cause infection. Moreover, SEM images of the treated cells exhibited the presence of wrinkles, deformities, and impaired cell walls, which suggests an alteration and instability of the membrane. This study demonstrated the efficacy of the greenly manufactured NPs in suppressing the proliferation of candida, drug-resistant bacteria, and their preexisting biofilms, as well as yeast-to-hyphae transformation. Therefore, these NPs with broad spectrum applications could be utilized in health settings to mitigate biofilm-related health conditions caused by pathogenic microbial strains.

Synthesis, in vitro and in silico screening of novel ferrocenyl substituted cyclohexenone and indazole derivatives as effective antimycobacterial agents

In response to the escalating threat of mycobacterial infections, including extensively drug-resistant tuberculosis (XDR) and multidrug-resistant tuberculosis (MDR), we synthesized novel ferrocene incorporating cyclohexenone and indazole derivatives. The compounds were synthesized via multistep reaction approach starting from chalcone precursors and evaluated against Mycobacterium fortuitum. The MIC and MBC values were calculated by following CLSI guidelines, with a focus on their ability to inhibit bacterial growth and disrupt biofilm formation. All the test compounds showed potent in vitro antimycobacterial activity against M. fortuitum. The MIC values range from 3.907 µg/mL to 500 µg/mL, whereas the MBC values range from 15.625 µg/mL to >500 µg/mL. Among the synthesized compounds, compound 3b exhibited potent antimycobacterial activity compared with standard drug Amikacin used to treat multidrug-resistant tuberculosis. 3b demonstrated significant inhibition of bacterial growth and showed the capacity to effectively suppress biofilm formation and disrupt pre-existing biofilms. This molecule also displayed favourable ADMET profile and good oral bioavailability, besides showing potential interactions with the target enzyme as revealed by docking studies and validated by enzyme assay. Collectively, these studies position 3b as a potential lead molecule that could be further optimized to develop ferrocene incorporating indazole based antimycobacterial agents.

Evaluating a polymicrobial biofilm model for structural components by co-culturing Komagataeibacter hansenii produced bacterial cellulose with Pseudomonas aeruginosa PAO1

A polymicrobial biofilm model of Komagataeibacter hansenii and Pseudomonas aeruginosa was developed to understand whether a pre-existing matrix affects the ability of another species to build a biofilm. P. aeruginosa was inoculated onto the preformed K. hansenii biofilm consisting of a cellulose matrix. P. aeruginosa PAO1 colonized and infiltrated the K. hansenii bacterial cellulose biofilm (BC), as indicated by the presence of cells at 19 μm depth in the translucent hydrogel matrix. Bacterial cell density increased along the imaged depth of the biofilm (17-19 μm). On day 5, the average bacterial count across sections was 67 ± 4 % P. aeruginosa PAO1 and 33 ± 6 % K. hansenii. Biophysical characterization of the biofilm indicated that colonization by P. aeruginosa modified the biophysical properties of the BC matrix, which inlcuded increased density, heterogeneity, degradation temperature and thermal stability, and reduced crystallinity, swelling ability and moisture content. This further indicates colonization of the biofilm by P. aeruginosa. While eDNA fibres - a key viscoelastic component of P. aeruginosa biofilm - were present on the surface of the co-cultured biofilm on day 1, their abundance decreased over time, and by day 5, no eDNA was observed, either on the surface or within the matrix. P. aeruginosa-colonized biofilm devoid of eDNA retained its mechanical properties. The observations demonstrate that a pre-existing biofilm scaffold of K. hansenii inhibits P. aeruginosa PAO1 eDNA production and suggest that eDNA production is a response by P. aeruginosa to the viscoelastic properties of its environment.

Hydroquinine Enhances the Efficacy of Contact Lens Solutions for Inhibiting Pseudomonas aeruginosa Adhesion and Biofilm Formation.

P. aeruginosa is one of the most common bacteria causing contact lens-related microbial keratitis (CLMK). Previous studies report that disinfecting solutions were ineffective in preventing biofilm formation. Solutions containing novel natural agents may be an excellent alternative for reducing the risk of CLMK. Here, we investigate the disinfecting properties of hydroquinine in combination with multipurpose solutions (MPSs) to prevent P. aeruginosa adhesion and biofilm formation. We examined the antibacterial, anti-adhesion, and anti-biofilm properties of hydroquinine-formulated MPSs compared to MPSs alone. Using RT-qPCR, hydroquinine directly affected the expression levels of adhesion-related genes, namely, cgrC, cheY, cheZ, fimU, and pilV, resulting in reduced adhesion and anti-biofilm formation. Using ISO 14729 stand-alone testing, hydroquinine met the criteria (>99.9% killing at disinfection time) against both P. aeruginosa reference and clinical strains. Using the crystal violet retention assay and FE-SEM, MPSs combined with hydroquinine were effective in inhibiting P. aeruginosa adhesion and destroying preexisting biofilms. This report is the first to highlight the potential utility of hydroquinine-containing formulations as a disinfecting solution for contact lenses, specifically for inhibiting adhesion and destroying biofilm. These findings may aid in the development of novel disinfectants aimed at combating P. aeruginosa, thereby potentially reducing the incidence of CLMK.

Antimicrobial and Antibiofilm Activity of Monolaurin against Methicillin-Resistant Staphylococcus aureus Isolated from Wound Infections.

Methicillin-resistant Staphylococcus aureus (MRSA) is one of the major pathogens associated with life-threatening infections, showing resistance to various antibiotics. This study aimed to assess the influence of monolaurin on biofilm-forming MRSA. The agar dilution method determined the minimum inhibitory concentration (MIC) of monolaurin against MRSA isolates and explored its impact on the resistance profile of selected antibiotics. The assessment of combined therapy involving monolaurin and antibiotics was conducted using fractional inhibitory concentration (FIC). The tissue culture plate strategy appraised monolaurin's antibiofilm activity and its inhibitory concentration (IC50), with assessment via scanning electron microscopy. Reverse transcription polymerase chain reaction (RT-PCR) discerned a monolaurin effect on the expression of the icaD gene. Monolaurin exhibited MIC values ranging from 500 to 2000 μg/mL. FIC index showed a synergistic effect of monolaurin with β-lactam antibiotics ranging from 0.0039 to 0.25 (p < 0.001). Among the 103 investigated MRSA isolates, 44 (44.7%) displayed moderate biofilm formation, while 59 (55.3%) were strong biofilm producers. Antibiofilm activity demonstrated concentration dependence, confirming monolaurin's capacity to inhibit biofilm formation and exhibited strong eradicating effects against preformed MRSA biofilms with IC50 values of 203.6 μg/mL and 379.3 μg/mL, respectively. Scanning electron microscope analysis revealed reduced cell attachments and diminished biofilm formation compared to the control. The expression levels of the icaD gene were remarkably reduced at monolaurin concentrations of 250 and 500 μg/mL. Monolaurin had significant inhibitory effects on MRSA pre-existing biofilms as well as biofilm development. So, it can be employed in the treatment of severe infections, particularly those associated with biofilm formation including catheter-associated infections.

Investigating the effect of Lactiplantibacillus plantarum TW57-4 in preventing biofilm formation and expression of virulence genes in Listeria monocytogenes ATCC 19115

Listeria monocytogenes is a common foodborne pathogen associated with listeriosis, a severe illness with a mortality rate of 20–30%. This study aimed to assess the antimicrobial properties of Lactiplantibacillus plantarum TW57-4 cell-free supernatant (CFS) against L. monocytogenes ATCC 19115. The CFS exhibited a minimum inhibitory concentration (MIC) of 31.24 mg/mL against L. monocytogenes. Scanning electron microscopy confirmed its anti-biofilm effects. The impact of CFS on various aspects of L. monocytogenes, such as biofilm formation, auto-aggregation, cytotoxicity, hemolysin activity, adhesion, and invasion, was also investigated. CFS treatment effectively inhibited biofilm formation at concentrations ranging from 1/16MIC (9.67%) to MIC (80%). It also reduced preexisting biofilms by 4.2%–70.91%. The CFS demonstrated cytotoxic effects on Caco-2 cancer cells (IC50, 44.64 mg/mL) and diminished hemolytic activity of L. monocytogenes at concentrations from MIC to 1/4 MIC, with reduction percentages of 50.66% and 28.66% respectively. Moreover, CFS treatment suppressed the expression of virulence genes, inhibiting the pathogenicity of L. monocytogenes. In conclusion, Lpb. plantarum TW57-4 exhibits favorable probiotic properties, antibacterial effects, and holds potential as a natural bacteriostatic agent. Further research and practical applications are warranted. It also shows promise as a bio-intervention for controlling L. monocytogenes biofilm in the food industry.

Pythium insidiosum: insights into biofilm formation and antibiofilm activity of antifungal drugs.

In this study, we investigate the ability of Pythium insidiosum to form biofilms across various substrates and the antibiofilm efficacy of 8-hydroxyquinoline derivatives (8-HQs). Biofilms of P. insidiosum were cultured on polystyrene plates, contact lenses, and horsehair. We provide the first evidence of P. insidiosum's biofilm-forming capability, thus considerably expanding our understanding of its transmission and pathogenesis. Our results demonstrate that 8-HQs effectively inhibit biofilm formation and eradicate pre-existing biofilms, underscoring their potential as a novel treatment strategy for pythiosis, a disease currently lacking a gold-standard treatment. This finding has particular relevance for ocular pythiosis associated with contact lens usage and potential infection sources in animals. Our results contribute to the scientific knowledge base and directly impact innovative therapeutic interventions' development.

Whole genome sequencing and functional analysis of a novel biofilm-eradicating strain Nocardiopsis lucentensis EMB25.

The process of biofilm formation is intricate and multifaceted, requiring the individual cells to secrete extracellular polymeric substances (EPS) that subsequently aggregate and adhere to various surfaces. The issue of biofilms is a significant concern for public health due to the increased resistance of microorganisms associated with biofilms to antimicrobial agents. The current study describes the whole genome and corresponding functions of a biofilm inhibiting and eradicating actinobacteria isolate identified as Nocardiopsis lucentensis EMB25. The N. lucentensis EMB25 has 6.5 Mbp genome with 71.62% GC content. The genome analysis by BLAST Ring Image Generator (BRIG) revealed it to be closely related to Nocardiopsis dassonvillei NOCA502F. Interestingly, based on orthologous functional groups reflected by average nucleotide identity (ANI) analysis, it was 81.48% similar to N. arvandica DSM4527. Also, it produces lanthipeptides and linear azole(in)e-containing peptides (LAPs) akin to N. arvandica. The secondary metabolite search revealed the presence of major gene clusters involved in terpene, ectoine, siderophores, Lanthipeptides, RiPP-like, and T1PKS biosynthesis. After 24h of treatment, the cell-free extract effectively eradicates the pre-existing biofilm of P. aeruginosa PseA. Also, the isolated bacteria exhibited antibacterial activity against MRSA, Staphylococcus aureus and Bacillus subtilis bacteria. Overall, this finding offers valuable insights into the identification of BGCs, which contain enzymes that play a role in the biosynthesis of natural products. Specifically, it sheds light on the functional aspects of these BGCs in relation to N. lucentensis.

Antipathogenic Efficacy of Biogenic Silver Nanoparticles and Antibiofilm Activities Against Multi-drug-Resistant ESKAPE Pathogens.

The silver nanoparticles (AgNPs) were produced by employing a biogenic loom and tested for antipathogenic assets against multi-drug-resistant (MDR) ESKAPE bacteria. Biogenically synthesized AgNPs were characterized adopting various high-throughput techniques such as UHRTEM, SEM with EDX, DLS, TGA-DTA, and XRD and spectroscopic analysis showed polydispersion of nanoparticles. In this context, AgNPs with the attribute of spherical-shaped nanoparticles with an average size of 26nm were successfully synthesized utilizing bacterial supernatant. The antipathogenic activities of AgNPs were assessed against 11 strains of MDR ESKAPE bacteria including Enterococcus faecium; methicillin-resistant Staphylococcus aureus; Klebsiella pneumonia; Acinetobacter baumannii; Pseudomonas aeruginosa; Enterobacter aerogenes; and Enterobacter species. The exposure of biogenic AgNPs in a well diffusion assay showed all the growth inhibitions of ESKAPE bacteria at 200μg/ml after 18h of incubation. Growth kinetics demonstrated maximum killing at 60μg/ml after 4h of completion. The highest biofilm depletions were found at 100μg/ml in adhesion assay. Live/dead assays showed effective killing of the ESKAPE bacteria at 10μg/ml in pre-existing biofilms. The effective inhibitory concentrations of AgNPs were investigated ranging from 10 to 200μg/ml. The selected pathogens found sensitive to AgNPs are statistically significant (P < 0.05) than that of cefotaxime/AgNO3. Consequently, a broad spectrum of antimicrobial potentials of AgNPs can be alternative to conventional antimicrobial agents for future medicine.

Role of Virus on Oral Biofilm: Inducer or Eradicator?

Sessile forms of bacteria remain as an aggregation on biotic and abiotic surfaces, known as biofilm, that protects them from various environmental stress, like antibiotic and host immune response. The oral cavity is enriched with microbial biofilm, formed on dental surface, gingival plaques, and associated tissue. Several pathogenic viruses enter the oral cavity and form biofilms either on pre-existing biofilms or on cell surfaces. They achieved persistence and the ability to prompt dissemination in the biofilm. Dental biofilms of COVID-19 patients are found to harbor SARS-CoV-2 RNA and may act as a budding reservoir, which also promotes COVID-19 transmission. On the other hand, most of the prokaryotic viruses or bacteriophages essentially kill the host bacteria and thereby destroy the biofilm. Bacteria try to evade from phage attack by concealing in biofilm, whereas the eukaryotic virus often utilize bacterial biofilm to escape host's immune response and to achieve an easy way of dissemination. The opposite action of viruses as an inducer and eradicator of biofilm has made the oral biofilm a unique ecosystem.

Piperine, a Plant Alkaloid, Exhibits Efficient Disintegration of the Pre-existing Biofilm of Staphylococcus aureus: a Step Towards Effective Management of Biofilm Threats.

Staphylococcus aureus causes a range of chronic infections in humans by exploiting its biofilm machinery and drug-tolerance property. Although several strategies have been proposed to eradicate biofilm-linked issues, here, we have explored whether piperine, a bioactive plant alkaloid, can disintegrate an already existing Staphylococcal biofilm. Towards this direction, the cells of S. aureus were allowed to develop biofilm first followed by treatment with the test concentrations (8 and 16µg/mL) of piperine. In this connection, several assays such as total protein recovery assay, crystal violet assay, extracellular polymeric substances (EPS) measurement assay, fluorescein diacetate hydrolysis assay, and fluorescence microscopic image analysis confirmed the biofilm-disintegrating property of piperine against S. aureus. Piperine reduced the cellular auto-aggregation by decreasing the cell surface hydrophobicity. On further investigation, we observed that piperine could down regulate the dltA gene expression that might reduce the cell surface hydrophobicity of S. aureus. It was also observed that the piperine-induced accumulation of reactive oxygen species (ROS) could enhance biofilm disintegration by decreasing the cell surface hydrophobicity of the test organism. Together, all the observations suggested that piperine could be used as a potential molecule for the effective management of the pre-existing biofilm of S. aureus.

Synergy with farnesol rejuvenates colistin activity against Colistin-resistant Gram-negative bacteria in vitro and in vivo

Colistin (COL) is considered the last line of treatment against infections due to multidrug-resistant (MDR) Gram-negative bacteria (GNB). However, the increasing number of colistin-resistant (COL-R) bacteria is a great threat to public health. In this study, a strategy of combining farnesol (FAR), which has anti-inflammatory and antitumor properties, with COL to restart COL activity was proposed. The synergistic effect of FAR combined with COL against COL-R GNB in vivo and in vitro were investigated. The excellent synergistic antibacterial activity of the COL–FAR combination was confirmed by performing the checkerboard assay, time-killing assay, and LIVE/DEAD bacterial cell viability assay. Crystal violet staining and scanning electron microscopy results showed that COL–FAR prevented biofilm formation and eradicated pre-existing mature biofilm. Cytotoxicity assay showed that FAR at 64 µg/mL was not cytotoxic to RAW264.7 cells. In vivo infection experiments showed that COL–FAR increased the survival rate of infected Galleria mellonella and decreased the bacterial load in a mouse thigh infection model. These results indicate that COL–FAR is a potentially effective therapeutic option for combating COL-R GNB infections.