Ines Sifaoui

Detection of Acanthamoeba on the ocular surface in a Spanish population using the Schirmer strip test: pathogenic potential, molecular classification and evaluation of the sensitivity to chlorhexidine and voriconazole of the isolated Acanthamoeba strains.

Pathogenic strains of Acanthamoeba are causative agents of a sight-threatening infection of the cornea known as Acanthamoeba keratitis, which is often associated with the misuse of contact lenses. However, there is still a question remaining to be answered, which is whether these micro-organisms are present on the ocular surface of healthy individuals. Therefore, the aim of this study was to determine the presence of Acanthamoeba on the ocular surface in healthy patients and also in those with other ocular surface infections. Sterile Schirmer test strips were used to collect samples from a group of patients who attended an ophthalmology consultation at the Hospital del Norte, Icod de los Vinos, Tenerife, Canary Islands. Most of the patients (46 individuals, 79.31  %) presented ocular surface pathologies such as blepharitis or conjunctivitis; the rest did not present any pathology. None of the patients included in the study wore contact lenses. The collected samples were cultured in 2  % non-nutrient agar plates and positive plates were then cultured in axenic conditions for further analyses. Molecular analysis classified all isolated strains as belonging to Acanthamoeba genotype tbl4, and osmotolerance and thermotolerance assays revealed that all strains were potentially pathogenic. Furthermore, all strains were assayed for sensitivity against voriconazole and chlorhexidine. Assays showed that both drugs were active against the tested strains. In conclusion, the Schirmer strip test is proposed as an effective tool for the detection of Acanthamoeba on the ocular surface.

Toxic effects of selected proprietary dry eye drops on Acanthamoeba

Amoebae of the genus Acanthamoeba are ubiquitous protists that have been isolated from many sources such as soils, water and the air. They are responsible for infections including fatal encephalitis and a severe keratitis in humans. To date, there is no satisfactorily effective therapeutic agent against this pathogen and the infections it causes are exacerbated by the existence of a resistant cyst stage produced by this amoeba. As dry eye syndrome is a risk factor for Acanthamoeba keratitis, we aimed to evaluate the anti-Acanthamoeba activity of a variety of proprietary eye drops intended to treat dry eye syndrome. From the nine eye drop formulations tested, “Systane Ultra” was determined to be the most active against all tested Acanthamoeba strains. During our investigations into the mode of action of Systane Ultra, we discovered that it decreases mitochondrial membrane potential and ATP levels, induces chromatin condensation, and increases the permeability of the plasma-membrane.

Therapeutic targets and investigated treatment strategies in Acanthamoeba keratitis

ABSTRACT Introduction: Free Living Amoebae (FLA) are unicellular protozoa common to most soil and aquatic environments. Only members of the genus Acanthamoeba and the species Naegleria fowleri, Balamuthia mandrillaris and Sappinia diploidea are known to infect humans and other animals, often with fatal consequences. Acanthamoeba species are the causative agents of a sight threating infection of the cornea known as Acanthamoeba keratitis (AK) that mainly affects immunocompetent contact lens wearers and has shown an increased incidence in the recent years worldwide. To date, most of the available treatments and contact lens maintenance solutions are not fully effective against these pathogens. Areas covered: There is a clear need for novel fully effective therapeutic agents against AK. In the recent years the application of ‘phylopharmacology’ and RNAi has shown to be a reliable tool for the elucidation of novel drug targets and therapeutic agents. Current therapeutic options for AK as well as ongoing research advances are presented and discussed. Expert opinion: The available AK treatments are not fully effective so far mostly due to the existence of a cyst stage in this pathogen. Moreover, most of the first line treatment options are not available in all countries. There is an urgent need to validate novel drug targets and therapeutic agents against AK. Recent approaches have allowed the validation of drug targets and novel compounds that could become fully effective therapies against these emerging pathogens.