Sodium Metabisulfite Inhibits Acanthamoeba Trophozoite Growth through Thiamine Depletion.

Acanthamoeba keratitis (AK) is a severe infection of the cornea. Prevention and treatment are difficult due to the inefficacy of currently available compounds. The impact of many commonly used compounds for routine examinations of Acanthamoeba is unexplored but might offer insight useful in combatting AK. In this study, we demonstrate that sodium metabisulfite, a common preservation constituent of eye care solutions, was found to be active against Acanthamoeba trophozoites at concentrations lower than that commonly found in eye drops (IC50 0.03 mg/mL). We demonstrate that sodium metabisulfite depletes thiamine from growth medium and that Acanthamoeba is a thiamine auxotroph, requiring thiamine salvage for growth. The inhibitory effects of sodium metabisulfite can be overcome by thiamine supplementation. These results are consistent with the lack of key enzymes for thiamine biosynthesis in the genome of Acanthamoeba, an area which might prove exploitable using new or existing compounds. Indeed, this study highlights sodium metabisulfite as a useful inhibitor of Acanthamoeba castellanii trophozoites in vitro and that it acts, at least in part, by limiting available thiamine.

The synergistic antibacterial activity of ozone and surfactant mists.

The microbiological safety of medical equipment and general surfaces is paramount to both the well-being of patients and the public. The application of ozone (a potent oxidant) has been recognised and implemented for this purpose, globally. However, it has primarily been utilised in the gaseous and aqueous forms. In this study, we investigate the potency of fine ozone mists and evaluate the synergistic effect when combined with cationic, anionic and non-ionic surfactants (dodecyl trimethyl ammonium bromide - DTAB, sodium dodecyl sulfate - SDS, alkyl polyglycoside - APG) as well as polyethylene glycol (PEG). Ozone mist is generated via a nebuliser (equipped with a compressed gas stream) and the piezoelectric method; whereas fabric substrates contaminated with Escherichia coli and Staphylococcus aureus are utilised in this study. Contamination levels on the fabric swatches are evaluated using agar dipslides. Compared to gaseous ozonation and aqueous ozonation (via nanobubble generation), the produced ozone mists showed significantly inferior antimicrobial properties for the tested conditions (6 ppm, 5-15 min). However, the hybrid mist-based application of 'ozone + surfactants' and 'ozone + PEG' showed considerable improvements compared to their independent applications (ozone mist only and surfactant mist only). The 'ozone + DTAB' mist had the highest activity, with better results observed with the micron-mist nebuliser than the piezoelectric transducer. We propose a likely mechanism for this synergistic performance (micellar encapsulation) and demonstrate the necessity for continued developments of novel decontamination technologies.

Alkyl-carbon chain length of two distinct compounds and derivatives are key determinants of their anti-Acanthamoeba activities.

The opportunistic pathogen, Acanthamoeba castellanii is the causative agent for the sight threatening infection Acanthamoeba keratitis (AK). It is commonly associated with contact lens wearers, and prevalence is increasing at an alarming rate due to an inadequate preventive strategy to protect the lens from this protist. This problem is compounded by the lack of an effective acanthamoebocide, particularly with cysticidal activity in the contact lens solutions. We have used cytotoxicity assays and a variety of biophysical approaches to show that two molecules with tails made of alkyl carbon, alkylphosphocholines (APCs) and quaternary ammonium compounds (QACs) had significant chain-length dependent efficacy against A. castellanii trophozoites, the latter producing death via permeabilization, and DNA complexing. QACs were more effective than APCs and had activity against cysts. Conversely, the QAC with 12 alkyl carbon chain, was non toxic, its presence increased A. castellanii trophozoites biomass and delayed encystation by 96 h. Interestingly, it was unable to induce excystation and increased trophozoite sensitivity to APC16. These results present a mono- and multi-inhibitor management strategy effective against trophozoites and cysts that may be useful for formulating into contact lense cleaning solutions and reducing AK incidence.