Letter to the Editor

Lysostaphin and clarithromycin: a promising combination for the eradication of Staphylococcus aureus biofilms

Although biofilm-specific antibiotic susceptibility assays are available, they are time-consuming and resource-intensive, and hence they are not usually performed in clinical settings. Herein, we introduce a machine learning-based predictive modeling approach that uses routinely available and easily accessible data to qualitatively predict in vitro antibiofilm activity of antibiotics with relatively high accuracy. Three optimized models based on logistic regression, decision tree, and random forest algorithms were successfully developed in this study using data manually collected from published literature. In these models, independent variables that serve as significant predictors of antibiofilm activity are minimum inhibitory concentration, bacterial Gram type, biofilm formation method, in addition to antibiotic's mechanism of action, molecular weight, and pKa. The cross-validation method showed that the optimized models exhibit prediction accuracy of 67% ± 6.1% for the logistic regression model, 73% ± 5.8% for the decision tree model, and 74% ± 5% for the random forest model. However, the one-way ANOVA test revealed that the difference in prediction accuracy between the 3 models is not statistically significant, and hence they can be considered to have comparable performance. The presented modeling approach can serve as an alternative to the resource-intensive biofilm assays to rapidly and properly manage biofilm-associated infections, especially in resource-limited clinical settings.

Microbial adhesion to surfaces and the subsequent formation of biofilms is a frequent problem encountered during food production. Biofilms allow microorganisms to survive in hostile environments. These microorganisms are then difficult to eradicate as their compact and complex structure hampers penetration and access to pathogenic cells, making them extremely resistant to stress conditions, particularly antimicrobials. The increasing prevalence of antibiotic resistance coupled with the ineffective control of conventional cleansing and disinfection regimes has generated a need for new, complementary, and/or alternative removal perspectives. Emerging strategies for the control of food quality and safety have thus arisen that involve the use of enzymes as innovative antibiofilm agents because of their ability to disintegrate extracellular polymer substances matrix components (microbial DNA, polysaccharides, and proteins). This chapter aims to address the problem of biofilms in the food industry and the different strategies employed to reduce or prevent their formation through the application of enzymes. Specifically, the types of enzymes, their mode of action, and the effects of combining enzymes with each other and/or with other antimicrobial techniques are described. Finally, the advantages and disadvantages of using enzymes as an antibiofilm strategy and future perspectives on the subject are discussed.

Staphylococcus aureus is a leading cause of healthcare-associated infections. The ability of S. aureus to attach and subsequently accumulate on the surfaces of implanted medical devices and in host tissues makes infections caused by this pathogen difficult to treat. Current treatments have been shown to have limited effect on surface-associated S. aureus, and may be enhanced by the addition of a dispersal agent. This study assessed the enzymatic agents dispersin B, lysostaphin, alpha amylase, V8 protease and serrapeptase, alone and in combination with vancomycin and rifampicin, against biofilms formed by meticillin-resistant and -susceptible strains of S. aureus. The efficacy of both antibiotics was enhanced when combined with any of the dispersal agents. Lysostaphin and serrapeptase were the most effective dispersal agents against all strains tested. These data indicate that combinations of biofilm dispersal agents and antibiotics may extend the therapeutic options for the treatment of S. aureus biofilm-associated infections.

Staphylococcus aureus infections of the skin and soft tissue pose a major concern to public health, largely owing to the steadily increasing prevalence of drug resistant isolates. As an alternative mode of treatment both bacteriophage endolysins and bacteriocins have been shown to possess antimicrobial efficacy against multiple species of bacteria including otherwise drug resistant strains. Despite this, the administration and exposure of such antimicrobials should be restricted until required in order to discourage the continued evolution of bacterial resistance, whilst maintaining the activity and stability of such proteinaceous structures. Utilising the increase in skin temperature during infection, the truncated bacteriophage endolysin CHAP_K and the staphylococcal bacteriocin lysostaphin have been co-administered in a thermally triggered manner from Poly(N-isopropylacrylamide) (PNIPAM) nanoparticles. The thermoresponsive nature of the PNIPAM polymer has been employed in order to achieve the controlled expulsion of a synergistic enzybiotic cocktail consisting of CHAP_K and lysostaphin. The point at which this occurs is modifiable, in this case corresponding to the threshold temperature associated with an infected wound. Consequently, bacterial lysis was observed at 37 °C, whilst growth was maintained at the uninfected skin temperature of 32 °C.

Cell wall hydrolases (CWH) are enzymes that build, remodel and degrade peptidoglycan within bacterial cell walls and serve essential roles in cell-wall metabolism, bacteriophage adsorption and bacteriolysis, environmental niche expansion, as well as eukaryotic innate immune defense against bacterial infection. Some CWHs, when tested as recombinant purified proteins, have been shown to have bactericidal activities both as single agents and in combinations with other antimicrobials, displaying synergies in vitro and potent activities in animal models of infection greater than the single agents alone. We summarize in vitro, in vivo, and mechanistic studies that illustrate ACWH synergy with antibiotics, antimicrobial peptides, and other ACWHs, underscoring the overall synergistic potential of the ACWH class.