Microbial Biosurfactants: Antimicrobial Activity and Potential Biomedical and Therapeutic Exploits

The rapid emergence of multidrug-resistant pathogens worldwide has raised concerns regarding the effectiveness of conventional antibiotics. This can be observed in ESKAPE pathogens, among others, whose multiple resistance mechanisms have led to a reduction in effective treatment options. Innovative strategies aimed at mitigating the incidence of antibiotic-resistant pathogens encompass the potential use of biosurfactants. These surface-active agents comprise a group of unique amphiphilic molecules of microbial origin that are capable of interacting with the lipidic components of microorganisms. Biosurfactant interactions with different surfaces can affect their hydrophobic properties and as a result, their ability to alter microorganisms’ adhesion abilities and consequent biofilm formation. Unlike synthetic surfactants, biosurfactants present low toxicity and high biodegradability and remain stable under temperature and pH extremes, making them potentially suitable for targeted use in medical and pharmaceutical applications. This review discusses the development of biosurfactants in biomedical and therapeutic uses as antimicrobial and antibiofilm agents, in addition to considering the potential synergistic effect of biosurfactants in combination with antibiotics. Furthermore, the anti-cancer and anti-viral potential of biosurfactants in relation to COVID-19 is also discussed

Assessment of Photoactivated Chlorophyllin Production of Singlet Oxygen and Inactivation of Foodborne Pathogens

Singlet oxygen (1O2) is known to have antibacterial activity; however, production can involve complex processes with expensive chemical precursors and/or significant energy input. Recent studies have confirmed the generation of 1O2 through the activation of photosensitizer molecules (PSs) with visible light in the presence of oxygen. Given the increase in the incidence of foodborne diseases associated with cross-contamination in food-processing industries, which is becoming a major concern, food-safe additives, such as chlorophyllins, have been studied for their ability to act as PSs. The fluorescent probe Singlet Oxygen Sensor Green (SOSG®) was used to estimate 1O2 formation upon the irradiation of traditional PSs (rose bengal (RB), chlorin 6 (ce6)) and novel chlorophyllins, sodium magnesium (NaChl) and sodium copper (NaCuChl), with both simulated-solar and visible light. NaChl gave rise to a similar 1O2 production rate when compared to RB and ce6. Basic mixing was shown to introduce sufficient oxygen to the PS solutions, preventing the limitation of the 1O2 production rate. The NaChl-based inactivation of Gram-positive S. aureus and Gram-negative E. coli was demonstrated with a 5-log reduction with UV–Vis light. The NaChl-based inactivation of Gram-positive S. aureus was accomplished with a 2-log reduction after 105 min of visible-light irradiation and a 3-log reduction following 150 min of exposure from an initial viable bacterial concentration of 106 CFU mL−1. CHS-NaChl-based photosensitization under visible light enhanced Gram-negative E. coli inactivation and provided a strong bacteriostatic effect preventing E. coli proliferation. The difference in the ability of NaChl and CHS-NaChl complexes to inactivate Gram-positive and Gram-negative bacteria was confirmed to result from the cell wall structure, which impacted PS–bacteria attachment and therefore the production of localized singlet oxygen

Biological and synthetic surfactant exposure increases antimicrobial gene occurrence in a freshwater mixed microbial biofilm environment

Publication history: Accepted - 8 March 2023; Published - 17 March 2023.Aquatic habitats are particularly susceptible to chemical pollution, such as antimicrobials, from domestic, agricultural, and industrial sources. This has led to the rapid increase of antimicrobial resistance (AMR) gene prevalence. Alternate approaches to counteract pathogenic bacteria are in development including synthetic and biological surfactants such as sodium dodecyl sulfate (SDS) and rhamnolipids. In the aquatic environment, these surfactants may be present as pollutants with the potential to affect biofilm formation and AMR gene occurrence. We tested the effects of rhamnolipid and SDS on aquatic biofilms in a freshwater stream in Northern Ireland. We grew biofilms on contaminant exposure substrates deployed within the stream over 4 weeks. We then extracted DNA and carried out shotgun sequencing using a MinION portable sequencer to determine microbial community composition, with 16S rRNA analyses (64,678 classifiable reads identified), and AMR gene occurrence (81 instances of AMR genes over 9 AMR gene classes) through a metagenomic analysis. There were no significant changes in community composition within all systems; however, biofilm exposed to rhamnolipid had a greater number of unique taxa as compared to SDS treatments and controls. AMR gene prevalence was higher in surfactant-treated biofilms, although not significant, with biofilm exposed to rhamnolipids having the highest presence of AMR genes and classes compared to the control or SDS treatments. Our results suggest that the presence of rhamnolipid encourages an increase in the prevalence of AMR genes in biofilms produced in mixed-use water bodies.EPSRC, Grant/Award Number: EP/P032427/1; University of Ulste

Biofilm regulation in <i>Clostridioides difficile</i>: Novel systems linked to hypervirulence

Clostridiodes difficile (C. difficile) was ranked an “urgent threat” by the Centers for Disease Control and Prevention (CDC) in 2019. C. difficile infection (CDI) is the most common healthcare-associated infection (HAI) in the United States of America as well as the leading cause of antibiotic-associated gastrointestinal disease. C. difficile is a gram-positive, rod-shaped, spore-forming, anaerobic bacterium that causes infection of the epithelial lining of the gut. CDI occurs most commonly after disruption of the human gut microflora following the prolonged use of broad-spectrum antibiotics. However, the recurrent nature of this disease has led to the hypothesis that biofilm formation may play a role in its pathogenesis. Biofilms are sessile communities of bacteria protected from extracellular stresses by a matrix of self-produced proteins, polysaccharides, and extracellular DNA. Biofilm regulation in C. difficile is still incompletely understood, and its role in disease recurrence has yet to be fully elucidated. However, many factors have been found to influence biofilm formation in C. difficile, including motility, adhesion, and hydrophobicity of the bacterial cells. Small changes in one of these systems can greatly influence biofilm formation. Therefore, the biofilm regulatory system would need to coordinate all these systems to create optimal biofilm-forming physiology under appropriate environmental conditions. The coordination of these systems is complex and multifactorial, and any analysis must take into consideration the influences of the stress response, quorum sensing (QS), and gene regulation by second messenger molecule cyclic diguanosine monophosphate (c-di-GMP). However, the differences in biofilm-forming ability between C. difficile strains such as 630 and the “hypervirulent” strain, R20291, make it difficult to assign a “one size fits all” mechanism to biofilm regulation in C. difficile. This review seeks to consolidate published data regarding the regulation of C. difficile biofilms in order to identify gaps in knowledge and propose directions for future study

Bobbin-Tool Friction-Stir Welding of Thick-Walled Aluminum Alloy Pressure Vessels

It was desired to assemble thick-walled Al alloy 2219 pressure vessels by bobbin-tool friction-stir welding. To develop the welding-process, mechanical-property, and fitness-for-service information to support this effort, extensive friction-stir welding-parameter studies were conducted on 2.5 cm. and 3.8 cm. thick 2219 Al alloy plate. Starting conditions of the plate were the fully-heat-treated (-T62) and in the annealed (-O) conditions. The former condition was chosen with the intent of using the welds in either the 'as welded' condition or after a simple low-temperature aging treatment. Since preliminary stress-analyses showed that stresses in and near the welds would probably exceed the yield-strength of both 'as welded' and welded and aged weld-joints, a post-weld solution-treatment, quenching, and aging treatment was also examined. Once a suitable set of welding and post-weld heat-treatment parameters was established, the project divided into two parts. The first part concentrated on developing the necessary process information to be able to make defect-free friction-stir welds in 3.8 cm. thick Al alloy 2219 in the form of circumferential welds that would join two hemispherical forgings with a 102 cm. inside diameter. This necessitated going to a bobbin-tool welding-technique to simplify the tooling needed to react the large forces generated in friction-stir welding. The bobbin-tool technique was demonstrated on both flat-plates and plates that were bent to the curvature of the actual vessel. An additional issue was termination of the weld, i.e. closing out the hole left at the end of the weld by withdrawal of the friction-stir welding tool. This was accomplished by friction-plug welding a slightly-oversized Al alloy 2219 plug into the termination-hole, followed by machining the plug flush with both the inside and outside surfaces of the vessel. The second part of the project involved demonstrating that the welds were fit for the intended service. This involved determining the room-temperature tensile and elastic-plastic fracture-toughness properties of the bobbin-tool friction-stir welds after a post-weld solution-treatment, quenching, and aging heat-treatment. These mechanical properties were used to conduct fracture-mechanics analyses to determine critical flaw sizes. Phased-array and conventional ultrasonic non-destructive examination was used to demonstrate that no flaws that match or exceed the calculated critical flaw-sizes exist in or near the friction-stir welds

Impact of a (poly)phenol-rich extract from the brown algae Ascophyllum nodosum on DNA damage and antioxidant activity in an overweight or obese population: a randomized controlled trial

Background Epidemiologic evidence suggests that a diet rich in (poly)phenols has beneficial effects on many chronic diseases. Brown seaweed is a rich source of (poly)phenols. Objective The aim of this study was to investigate the bioavailability and effect of a brown seaweed (Ascophyllum nodosum) (poly)phenol extract on DNA damage, oxidative stress, and inflammation in vivo. Design A randomized, double-blind, placebo-controlled crossover trial was conducted in 80 participants aged 30–65 y with a body mass index (in kg/m2) ≥25. The participants consumed either a 400-mg capsule containing 100 mg seaweed (poly)phenol and 300 mg maltodextrin or a 400-mg maltodextrin placebo control capsule daily for an 8-wk period. Bioactivity was assessed with a panel of blood-based markers including lymphocyte DNA damage, plasma oxidant capacity, C-reactive protein (CRP), and inflammatory cytokines. To explore the bioavailability of seaweed phenolics, an untargeted metabolomics analysis of urine and plasma samples after seaweed consumption was determined by ultra-high-performance liquid chromatography–high-resolution mass spectrometry. Results Consumption of the seaweed (poly)phenols resulted in a modest decrease in DNA damage but only in a subset of the total population who were obese. There were no significant changes in CRP, antioxidant status, or inflammatory cytokines. We identified phlorotannin metabolites that are considered potential biomarkers of seaweed consumption including pyrogallol/phloroglucinol-sulfate, hydroxytrifurahol A-glucuronide, dioxinodehydroeckol-glucuronide, diphlorethol sulfates, C-O-C dimer of phloroglucinol sulfate, and C-O-C dimer of phloroglucinol. Conclusions To the best of our knowledge, this work represents the first comprehensive study investigating the bioactivity and bioavailability of seaweed (poly)phenolics in human participants. We identified several potential biomarkers of seaweed consumption. Intriguingly, the modest improvements in DNA damage were observed only in the obese subset of the total population. The subgroup analysis should be considered exploratory because it was not preplanned; therefore, it was not powered adequately. Elucidation of the biology underpinning this observation will require participant stratification according to weight in future studies. This trial was registered at clinicaltrials.gov as NCT02295878

Inactivation of the dnaK gene in Clostridium difficile 630 Δerm yields a temperature-sensitive phenotype and increases biofilm-forming ability

Abstract Clostridium difficile infection is a growing problem in healthcare settings worldwide and results in a considerable socioeconomic impact. New hypervirulent strains and acquisition of antibiotic resistance exacerbates pathogenesis; however, the survival strategy of C. difficile in the challenging gut environment still remains incompletely understood. We previously reported that clinically relevant heat-stress (37–41 °C) resulted in a classical heat-stress response with up-regulation of cellular chaperones. We used ClosTron to construct an insertional mutation in the dnaK gene of C. difficile 630 Δerm. The dnaK mutant exhibited temperature sensitivity, grew more slowly than C. difficile 630 Δerm and was less thermotolerant. Furthermore, the mutant was non-motile, had 4-fold lower expression of the fliC gene and lacked flagella on the cell surface. Mutant cells were some 50% longer than parental strain cells, and at optimal growth temperatures, they exhibited a 4-fold increase in the expression of class I chaperone genes including GroEL and GroES. Increased chaperone expression, in addition to the non-flagellated phenotype of the mutant, may account for the increased biofilm formation observed. Overall, the phenotype resulting from dnaK disruption is more akin to that observed in Escherichia coli dnaK mutants, rather than those in the Gram-positive model organism Bacillus subtilis

Increased sporulation underpins adaptation of Clostridium difficile strain 630 to a biologically–relevant faecal environment, with implications for pathogenicity

Abstract Clostridium difficile virulence is driven primarily by the processes of toxinogenesis and sporulation, however many in vitro experimental systems for studying C. difficile physiology have arguably limited relevance to the human colonic environment. We therefore created a more physiologically–relevant model of the colonic milieu to study gut pathogen biology, incorporating human faecal water (FW) into growth media and assessing the physiological effects of this on C. difficile strain 630. We identified a novel set of C. difficile–derived metabolites in culture supernatants, including hexanoyl– and pentanoyl–amino acid derivatives by LC-MSn. Growth of C. difficile strain 630 in FW media resulted in increased cell length without altering growth rate and RNA sequencing identified 889 transcripts as differentially expressed (p < 0.001). Significantly, up to 300–fold increases in the expression of sporulation–associated genes were observed in FW media–grown cells, along with reductions in motility and toxin genes’ expression. Moreover, the expression of classical stress–response genes did not change, showing that C. difficile is well–adapted to this faecal milieu. Using our novel approach we have shown that interaction with FW causes fundamental changes in C. difficile biology that will lead to increased disease transmissibility