Modulation of Biofilm Growth by Sub‐Inhibitory Amounts of Antibacterial Substances

It is generally accepted that bacteria in biofilm are more resistant to antibacterials than their planktonic counterparts. For numerous antibiotics, it has been shown that minimal inhibitory concentrations (MICs) for bacteria grown in broth are much lower than the minimal biofilm inhibition concentrations. While sub‐inhibitory concentrations, that is, amounts of antibacterials below the MIC, do not either influence or suppress to some extent or other the bacterial growth in liquid media, these same amounts of drugs, natural substances, etc., may have diverse effects on bacterial biofilms, ranging from suppression to stimulation of the sessile growth and varying with regard to the bacterial species and strains. This is a source of additional risks for both biofilm infection of host tissues and contamination indwelling devices. When considering the data for biofilm modulation, differences in experimental protocols should be taken into account, as well as the strain‐specific mechanisms of biofilm formation

Plastic Degradation by Extremophilic Bacteria

Intensive exploitation, poor recycling, low repeatable use, and unusual resistance of plastics to environmental and microbiological action result in accumulation of huge waste amounts in terrestrial and marine environments, causing enormous hazard for human and animal life. In the last decades, much scientific interest has been focused on plastic biodegradation. Due to the comparatively short evolutionary period of their appearance in nature, sufficiently effective enzymes for their biodegradation are not available. Plastics are designed for use in conditions typical for human activity, and their physicochemical properties roughly change at extreme environmental parameters like low temperatures, salt, or low or high pH that are typical for the life of extremophilic microorganisms and the activity of their enzymes. This review represents a first attempt to summarize the extraordinarily limited information on biodegradation of conventional synthetic plastics by thermophilic, alkaliphilic, halophilic, and psychrophilic bacteria in natural environments and laboratory conditions. Most of the available data was reported in the last several years and concerns moderate extremophiles. Two main questions are highlighted in it: which extremophilic bacteria and their enzymes are reported to be involved in the degradation of different synthetic plastics, and what could be the impact of extremophiles in future technologies for resolving of pollution problems

The concept for the antivirulence therapeutics approach as alternative to antibiotics: hope or still a fiction?

AbstractWhile the development of antibiotics since their first discovery brought about a revolutionary step forward in the fight against infectious microorganisms, unfortunately its side effect was the highly increasing risks of antibiotic resistance. Resistance development poses the urgent task for discovery of novel prospective approaches in the fight against multidrug resistant bacteria. Together with the search for new antibacterials, there is a growing interest in novel non-traditional approaches. Such non-traditional approaches are the attempts to suppress bacterial virulence and the development of virulence-related phenotypes, instead of killing the bacteria. The focus of this review falls on the bacterial regulatory mechanisms of virulence expression via quorum sensing (QS), and the formation of multicellular communities—biofilms, that protect bacteria from the host defenses and the antibacterial substances. The review gives a general outline of two types of approaches for control of bacterial virulence-related phenotypes. One is the search for reagents with expected antivirulence efficacy via the inhibition of QS, for example among low molecular weight metabolites of different medicinal plants. The other is directed to the possible prevention and/or destruction of bacterial biofilms, which are a well-recognized source of chronic, persistent and recurrent infections. The concerns regarding possible practical applications are considered as well

Effects of cationic polymers on the viability of microbial biofilms

Introduction: The number of published biofilm studies and novel ways for studying them has risen dramatically in recent years, owing to the broad application of biofilms in medicine. Some bacteria develop biofilms that are highly resistant to antimicrobial agents, resulting in persistent infections. This necessitates the development of alternative methods for combating biofilms. In this regard, the application of cationic polymers is a good candidate for realization of this strategy. Aim: The aim of our study was to investigate the potential of a newly synthesized covalently attached star copolymer of N,N’-dimethylaminoethyl methacrylate and hydroxyl-bearing oligo(ethylene glycol) methacrylate [P(DMAEMA-co-HOEGMA)] to silica surfaces and its quaternized version [P(QDMAEMA-co-HOEGMA)] for destruction of biofilms formed by Bacillus subtilis or Pseudomonas aeruginosa. Materials and methods: Model strains representing different genera and taxonomic groups were selected for the study. The anti-biofilm activities of two different newly synthesized cationic polymers were investigated by observation (live/dead staining) of the viability of bacterial cells within the biofilm. Results: The results obtained by the live/dead labeling of bacterial biofilms show a substantial decrease in the viability of population in the presence of cationic polymers, better expressed at B. subtilis. Conclusions: The studied two immobilized on silica wafers newly synthesized star copolymers exhibited potential for anti-biofilm effects. The results demonstrated combined potential for reducing the viability of bacterial cells within the biofilms and probably for loosening the biofilm matrix. The effect was better expressed in B. subtilis

Ciprofloxacin-Loaded Mixed Polymeric Micelles as Antibiofilm Agents

In this work, mixed polymeric micelles (MPMs) based on a cationic poly(2-(dimethylamino)ethyl methacrylate)-b-poly(ε-caprolactone)-b-poly(2-(dimethylamino)ethyl methacrylate) (PDMAEMA29-b-PCL70-b-PDMAEMA29) and a non-ionic poly(ethylene oxide)–b-poly(propylene oxide)–b-poly(ethylene oxide) (PEO99-b-PPO67-b-PEO99) triblock copolymers, blended at different molar ratios, were developed. The key physicochemical parameters of MPMs, including size, size distribution, and critical micellar concentration (CMC), were evaluated. The resulting MPMs are nanoscopic with a hydrodynamic diameter of around 35 nm, and the ζ-potential and CMC values strongly depend on the MPM’s composition. Ciprofloxacin (CF) was solubilized by the micelles via hydrophobic interaction with the micellar core and electrostatic interaction between the polycationic blocks, and the drug localized it, to some extent, in the micellar corona. The effect of a polymer-to-drug mass ratio on the drug-loading content (DLC) and encapsulation efficiency (EE) of MPMs was assessed. MPMs prepared at a polymer-to-drug mass ratio of 10:1 exhibited very high EE and a prolonged release profile. All micellar systems demonstrated their capability to detach pre-formed Gram-positive and Gram-negative bacterial biofilms and significantly reduced their biomass. The metabolic activity of the biofilm was strongly suppressed by the CF-loaded MPMs indicating the successful drug delivery and release. The cytotoxicity of empty and CF-loaded MPMs was evaluated. The test reveals composition-dependent cell viability without cell destruction or morphological signs of cell death

Effects of Aromatic Compounds Degradation on Bacterial Cell Morphology

The aim of the present study was to evaluate in parallel the capacity of three bacterial strains originating from oil-polluted soils to degrade monoaromatic compounds and the alterations in the bacterial cell morphology as a result of the biodegradation. The strain Gordonia sp. 12/5 can grow well in media containing catechol, o-, m-, and p-cresol without significant morphological changes in the cells, as shown by scanning electron microscopy. This implies good adaptation of the strain for growth in hydrocarbon-containing media and indicates it is a proper candidate strain for further development of purification methodologies applicable to ecosystems contaminated with such compounds. The growth of the two Rhodococcus strains in the presence of the above carbon sources is accompanied by changes in cell size characteristic of stress conditions. Nevertheless, their hydrocarbon-degrading capacity should not be neglected for future applications. In summary, the established ability to degrade monoaromatic compounds, in parallel with the morphological changes of the bacterial cells, can be used as a valuable indicator of the strain’s vitality in the presence of tested aromatic compounds and, accordingly, of its applicability for bioremediation purposes

An Overview of Biofilm-Associated Infections and the Role of Phytochemicals and Nanomaterials in Their Control and Prevention

Biofilm formation is considered one of the primary virulence mechanisms in Gram-positive and Gram-negative pathogenic species, particularly those responsible for chronic infections and promoting bacterial survival within the host. In recent years, there has been a growing interest in discovering new compounds capable of inhibiting biofilm formation. This is considered a promising antivirulence strategy that could potentially overcome antibiotic resistance issues. Effective antibiofilm agents should possess distinctive properties. They should be structurally unique, enable easy entry into cells, influence quorum sensing signaling, and synergize with other antibacterial agents. Many of these properties are found in both natural systems that are isolated from plants and in synthetic systems like nanoparticles and nanocomposites. In this review, we discuss the clinical nature of biofilm-associated infections and some of the mechanisms associated with their antibiotic tolerance. We focus on the advantages and efficacy of various natural and synthetic compounds as a new therapeutic approach to control bacterial biofilms and address multidrug resistance in bacteria

Polyphasic characterisation of Microcoleus autumnalis (Gomont, 1892) Strunecky, Komárek & J.R.Johansen, 2013 (Oscillatoriales, Cyanobacteria) using a metabolomic approach as a complementary tool

As a result of the continuous revision of cyanobacterial taxonomy, Phormidium autumnale (Agardh) Trevisan ex Gomont, 1892 has been transferred to the genus Microcoleus as Microcoleus autumnalis (Gomont, 1892) Strunecky, Komárek & J.R.Johansen, 2013. This transfer was based on a single strain and literature data. In the present study, we revise the taxonomic position of Microcoleus autumnalis by applying the classical approach of polyphasic taxonomy and additionally using metabolomics. Cyanobacterial strains identified as Phormidium autumnale and Microcoleus vaginatus (type species of the genus Microcoleus) were used for comparative analyses. In addition, the taxonomic relationship between the species Phormidium autumnale and Phormidium uncinatum was determined on the basis of polyphasic characteristics. Monitoring of the morphological variability of Phormidium autumnale and Microcoleus vaginatus strains showed a difference in the morphology concerning the ends of the trichomes, the shape of the apical cells, as well as the presence/absence of the calyptra and its shape. The performed TEM analysis of the thylakoid arrangement of the studied strains showed parietal arrangement of the thylakoids in the representatives of genus Phormidium and fascicular arrangement in genus Microcoleus. Molecular genetic analyses, based on 16S rDNA, revealed grouping of the investigated P. autumnale strains in a separate clade. This clade is far from the subtree, which is very clearly formed by the representatives of the type species of genus Microcoleus, namely M. vaginatus. The metabolomic analysis involving P. autumnale and M. vaginatus strains identified 39 compounds that could be used as potential biochemical markers to distinguish the two cyanobacterial species. Based on the data obtained, we suggest changing of the current status of Microcoleus autumnalis by restoring its previous appurtenance to the genus Phormidium under the name Phormidium autumnale (Agardh) Trevisan ex Gomont, 1892 and distinguishing this species from genus Microcoleus

Structural, Thermal, and Storage Stability of Rapana Thomasiana Hemocyanin in the Presence of Cholinium-Amino Acid-Based Ionic Liquids

Novel biocompatible compounds that stabilize proteins in solution are in demand for biomedical and/or biotechnological applications. Here, we evaluated the effect of six ionic liquids, containing mono- or dicholinium [Chol]1or2 cation and anions of charged amino acids such as lysine [Lys], arginine [Arg], aspartic acid [Asp], or glutamic acid [Glu], on the structure, thermal, and storage stability of the Rapana thomasiana hemocyanin (RtH). RtH is a protein with huge biomedicinal potential due to its therapeutic, drug carrier, and adjuvant properties. Overall, the ionic liquids (ILs) induce changes in the secondary structure of RtH. However, the structure near the Cu-active site seems unaltered and the oxygen-binding capacity of the protein is preserved. The ILs showed weak antibacterial activity when tested against three Gram-negative and three Gram-positive bacterial strains. On the contrary, [Chol][Arg] and [Chol][Lys] exhibited high anti-biofilm activity against E. coli 25213 and S. aureus 29213 strains. In addition, the two ILs were able to protect RtH from chemical and microbiological degradation. Maintained or enhanced thermal stability of RtH was observed in the presence of all ILs tested, except for RtH-[Chol]2[Glu]