Antimicrobial and anti-biofilm potencies of dermcidin-derived peptide DCD-1L against Acinetobacter baumannii: an in vivo wound healing model

Background The global emergence of Acinetobacter baumannii resistance to most conventional antibiotics presents a major therapeutic challenge and necessitates the discovery of new antibacterial agents. The purpose of this study was to investigate in vitro and in vivo anti-biofilm potency of dermcidin-1L (DCD-1L) against extensively drug-resistant (XDR)-, pandrug-resistant (PDR)-, and ATCC19606-A. baumannii. Methods After determination of minimum inhibitory concentration (MIC) of DCD-1L, in vitro anti-adhesive and anti-biofilm activities of DCD-1L were evaluated. Cytotoxicity, hemolytic activity, and the effect of DCD-1L treatment on the expression of various biofilm-associated genes were determined. The inhibitory effect of DCD-1L on biofilm formation in the model of catheter-associated infection, as well as, histopathological examination of the burn wound sites of mice treated with DCD-1L were assessed. Results The bacterial adhesion and biofilm formation in all A. baumannii isolates were inhibited at 2 × , 4 × , and 8 × MIC of DCD-1L, while only 8 × MIC of DCD-1L was able to destroy the pre-formed biofilm in vitro. Also, reduce the expression of genes involved in biofilm formation was observed following DCD-1L treatment. DCD-1L without cytotoxic and hemolytic activities significantly reduced the biofilm formation in the model of catheter-associated infection. In vivo results showed that the count of A. baumannii in infected wounds was significantly decreased and the promotion in wound healing by the acceleration of skin re-epithelialization in mice was observed following treatment with 8 × MIC of DCD-1L. Conclusions Results of this study demonstrated that DCD-1L can inhibit bacterial attachment and biofilm formation and prevent the onset of infection. Taking these properties together, DCD-1L appears as a promising candidate for antimicrobial and anti-biofilm drug development.

A metallic anti-biofouling surface with a hierarchical topography containing nanostructures on curved micro-riblets

Metallic surface finishes have been used in the anti-biofouling, but it is very difficult to produce surfaces with hierarchically ordered structures. In the present study, anti-biofouling metallic surfaces with nanostructures superimposed on curved micro-riblets were produced via top-down fabrication. According to the attachment theory, these surfaces feature few attachment points for organisms, the nanostructures prevent the attachment of bacteria and algal zoospores, while the micro-riblets prohibit the settlement of macrofoulers. Anodic oxidation was performed to induce superhydrophilicity. It forms a hydration layer on the surface, which physically blocks foulant adsorption along with the anti-biofouling topography. We characterized the surfaces via scanning electron and atomic force microscopy, contact-angle measurement, and wear-resistance testing. The contact angle of the hierarchical structures was less than 1°. Laboratory settlement assays verified that bacterial attachment was dramatically reduced by the nanostructures and/or the hydration layer, attributable to superhydrophilicity. The micro-riblets prohibited the settlement of macrofoulers. Over 77 days of static immersion in the sea during summer, the metallic surface showed significantly less biofouling compared to a surface painted with an anticorrosive coating.

The lolB gene in Xanthomonas campestris pv. campestris is required for bacterial attachment, stress tolerance, and virulence

Background Xanthomonas campestris pv. campestris (Xcc) is a Gram-negative bacterium that can cause black rot disease in crucifers. The lipoprotein outer membrane localization (Lol) system is involved in the lipoprotein sorting to the outer membrane. Although Xcc has a set of annotated lol genes, there is still little known about the physiological role in this phytopathogen. In this study, we aimed to characterize the role of LolB of Xcc in bacterial attachment, stress tolerance, and virulence. Results To characterize the role of LolB, lolB mutant was constructed and phenotypic evaluation was performed. The lolB mutant revealed reductions in bacterial attachment, extracellular enzyme production, and virulence. Mutation of lolB also resulted in reduced tolerance to a myriad of stresses, including heat and a range of membrane-perturbing agents. Trans-complementation of lolB mutant with intact lolB gene reverted these altered phenotypes to the wild-type levels. From subsequent reporter assay and reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) analysis, the expression of genes that encode the major extracellular enzymes and the stress-related proteins was reduced after lolB mutation. Conclusions The results in this work contribute to the functional understanding of lolB in Xanthomonas for the first time, and provide new insights into the function of lolB in bacteria.

Control of Multidrug-Resistant Pathogenic Staphylococci Associated with Vaginal Infection Using Biosurfactants Derived from Potential Probiotic Bacillus Strain

Biosurfactants exhibit antioxidant, antibacterial, antifungal, and antiviral activities. They can be used as therapeutic agents and in the fight against infectious diseases. Moreover, the anti-adhesive properties against several pathogens point to the possibility that they might serve as an anti-adhesive coating agent for medical inserts and prevent nosocomial infections, without using synthetic substances. In this study, the antimicrobial, antibiofilm, cell surface hydrophobicity, and antioxidative activities of biosurfactant extracted from Bacillus sp., against four pathogenic strains of Staphylococcus spp. associated with vaginal infection, were studied. Our results have shown that the tested biosurfactant possesses a promising antioxidant potential, and an antibacterial potency against multidrug clinical isolates of Staphylococcus, with an inhibitory diameter ranging between 27 and 37 mm, and a bacterial growth inhibition at an MIC of 1 mg/ mL, obtained. The BioSa3 was highly effective on the biofilm formation of different tested pathogenic strains. Following their treatment by BioSa3, a significant decrease in bacterial attachment (p < 0.05) was justified by the reduction in the optical (from 0.709 to 0.111) following their treatment by BioSa3. The antibiofilm effect can be attributed to its ability to alter the membrane physiology of the tested pathogens to cause a significant decrease (p < 0.05) of over 50% of the surface hydrophobicity. Based on the obtained result of the bioactivities in the current study, BioSa3 is a good candidate in new therapeutics to better control multidrug-resistant bacteria and overcome bacterial biofilm-associated infections by protecting surfaces from microbial contamination.

Facilitating GL13K Peptide Grafting on Polyetheretherketone via 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide: Surface Properties and Antibacterial Activity

In the present work, the antimicrobial peptide (AMP) of GL13K was successfully coated onto a polyetheretherketone (PEEK) substrate to investigate its antibacterial activities against Staphylococcus aureus (S. aureus) bacteria. To improve the coating efficiency, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) was mixed with a GL13K solution and coated on the PEEK surface for comparison. Both energy-dispersive X-ray spectroscopy (EDX) and X-ray photoelectron spectroscopy (XPS) data confirmed 30% greater peptide coating on PEEK/GL13K-EDC than PEEK without EDC treatment. The GL13K graft levels are depicted in the micrograms per square centimeter range. The PEEK/GL13K-EDC sample showed a smoother and lower roughness (Rq of 0.530 µm) than the PEEK/GL13K (0.634 µm) and PEEK (0.697 µm) samples. The surface of the PEEK/GL13K-EDC was more hydrophilic (with a water contact angle of 24°) than the PEEK/GL13K (40°) and pure PEEK (89°) samples. The pure PEEK disc did not exhibit any inhibition zone against S. aureus. After peptide coating, the samples demonstrated significant zones of inhibition: 28 mm and 25 mm for the PEEK/GL13K-EDC and PEEK/GL13K samples, respectively. The bacteria-challenged PEEK sample showed numerous bacteria clusters, whereas PEEK/GL13K contained a little bacteria and PEEK/GL13K-EDC had no bacterial attachment. The results confirm that the GL13K peptide coating was able to induce antibacterial and biofilm-inhibitory effects. To the best of our knowledge, this is the first report of successful GL13K peptide grafting on a PEEK substrate via EDC coupling. The present work illustrates a facile and promising coating technique for a polymeric surface to provide bactericidal activity and biofilm resistance to medical implantable devices.

Strategies for Antimicrobial Peptides Immobilization on Surfaces to Prevent Biofilm Growth on Biomedical Devices

Nosocomial and medical device-induced biofilm infections affect millions of lives and urgently require innovative preventive approaches. These pathologies have led to the development of numerous antimicrobial strategies, an emergent topic involving both natural and synthetic routes, among which some are currently under testing for clinical approval and use. Antimicrobial peptides (AMPs) are ideal candidates for this fight. Therefore, the strategies involving surface functionalization with AMPs to prevent bacterial attachment/biofilms formation have experienced a tremendous development over the last decade. In this review, we describe the different mechanisms of action by which AMPs prevent bacterial adhesion and/or biofilm formation to better address their potential as anti-infective agents. We additionally analyze AMP immobilization techniques on a variety of materials, with a focus on biomedical applications. Furthermore, we summarize the advances made to date regarding the immobilization strategies of AMPs on various surfaces and their ability to prevent the adhesion of various microorganisms. Progress toward the clinical approval of AMPs in antibiotherapy is also reviewed.

In situ control of root–bacteria interactions using optical trapping in transparent soil

Bacterial attachment on root surfaces is an important step preceding the colonisation or internalisation and subsequent infection of plants by pathogens. Unfortunately, bacterial attachment is not well understood because the phenomenon is difficult to observe. Here we assessed whether this limitation could be overcome using optical trapping approaches. We have developed a system based on counter-propagating beams and studied its ability to guide Pectobacterium atrosepticum (Pba) cells to different root cell types within the interstices of transparent soils. Bacterial cells were successfully trapped and guided to root hair cells, epidermis cells, border cells and tissues damaged by laser ablation. Finally, we used the system to quantify the bacterial cell detachment rate of Pba cells on root surfaces following reversible attachment. Optical trapping techniques could greatly enhance our ability to deterministically characterise mechanisms linked to attachment and formation of biofilms in the rhizosphere.

Inhibition of Bacterial Adhesion and Antibiofilm Activities of a Glycolipid Biosurfactant from Lactobacillus rhamnosus with Its Physicochemical and Functional Properties

Biosurfactants derived from different microbes are an alternative to chemical surfactants, which have broad applications in food, oil, biodegradation, cosmetic, agriculture, pesticide and medicine/pharmaceutical industries. This is due to their environmentally friendly, biocompatible, biodegradable, effectiveness to work under various environmental conditions and non-toxic nature. Lactic acid bacteria (LAB)-derived glycolipid biosurfactants can play a major role in preventing bacterial attachment, biofilm eradication and related infections in various clinical settings and industries. Hence, it is important to explore and identify the novel molecule/method for the treatment of biofilms of pathogenic bacteria. In the present study, a probiotic Lactobacillus rhamnosus (L. rhamnosus) strain was isolated from human breast milk. Firstly, its ability to produce biosurfactants, and its physicochemical and functional properties (critical micelle concentration (CMC), reduction in surface tension, emulsification index (% EI24), etc.) were evaluated. Secondly, inhibition of bacterial adhesion and biofilm eradication by cell-bound biosurfactants from L. rhamnosus was performed against various biofilm-forming pathogens (B. subtilis, P. aeruginosa, S. aureus and E. coli). Finally, bacterial cell damage, viability of cells within the biofilm, exopolysaccharide (EPS) production and identification of the structural analogues of the crude biosurfactant via gas chromatography–mass spectrometry (GC–MS) analysis were also evaluated. As a result, L. rhamnosus was found to produce 4.32 ± 0.19 g/L biosurfactant that displayed a CMC of 3.0 g/L and reduced the surface tension from 71.12 ± 0.73 mN/m to 41.76 ± 0.60 mN/m. L. rhamnosus cell-bound crude biosurfactant was found to be effective against all the tested bacterial pathogens. It displayed potent anti-adhesion and antibiofilm ability by inhibiting the bacterial attachment to surfaces, leading to the disruption of biofilm formation by altering the integrity and viability of bacterial cells within biofilms. Our results also confirm the ability of the L. rhamnosus cell-bound-derived biosurfactant to damage the architecture of the biofilm matrix, as a result of the reduced total EPS content. Our findings may be further explored as a green alternative/approach to chemically synthesized toxic antibiofilm agents for controlling bacterial adhesion and biofilm eradication.

Durability Assessment of a Plasma-Polymerized Coating with Anti-Biofilm Activity against L. monocytogenes Subjected to Repeated Sanitization

Biofilm formation on food-contact surfaces is a matter of major concern causing food safety and spoilage issues to this sector. The aim of this study was to assess the durability of the anti-biofilm capacity of a plasma-polymerized coating composed of a base coating of (3-aminopropyl)triethoxysilane (APTES) and a functional coating of acrylic acid (AcAc). Coated and uncoated AISI 316 stainless steel (SS) plates were subjected to five sanitization cycles with sodium hypochlorite (0.05%) and peracetic acid (0.5%). The effectiveness of the coating for the inhibition of multi-strain Listeria monocytogenes biofilm formation was confirmed using a three-strain cocktail, which was grown on the SS plates at 12 °C for 6 days. Compared to the uncoated SS, relative biofilm productions of 14.6% on the non-sanitized coating, 27.9% on the coating after sanitization with sodium hypochlorite, and 82.3% on the coating after sanitization with peracetic acid were obtained. Morphological and physicochemical characterization of the coatings suggested that the greater anti-biofilm effectiveness after sanitization with sodium hypochlorite was due to the high pH of this solution, which caused a deprotonation of the carboxylic acid groups of the functional coating. This fact conferred it a strong hydrophilicity and negatively charged its surface, which was favorable for preventing bacterial attachment and biofilm formation.