RESUMEN
INTRODUCTION: Several strains of the free-living genus Acanthamoeba can cause granulomatous amoebic encephalitis (GAE), a rare chronic and slowly progressive infection of the central nervous system (CNS), and Acanthamoeba keratitis (AK), a sight-threatening eye infectious disease. AK incidence has increased with the popularization of the contact lens wear and its treatment is currently limited and frequently unsuccessful. As imidazolium salts (IS), cationic imidazole derivatives, have promising antimicrobial potential. MATERIALS AND METHODS: The present study evaluated the amoebicidal activity of four IS; 1-n-hexadecyl-3-methylimidazolium methanesulfonate (C16MImMeS), chloride (C16MImCl) and bis (triluoromethylsulfonyl) imide (C16MImNTf2 ), and 1-methyl-3-n-octadecylimidazolium chloride (C18MImCl), against the Acanthamoeba castellanii (ATCC30010) environmental strain and a clinical isolate (genotype T4). RESULTS: Three IS showed being lethal to 100% of the Acanthamoeba trophozoites at the minimum inhibitory concentrations of 125 and 62.5 µg/mL (C16MImMeS), 31.25 and 62.5 µg/mL (C16MImCl), and 125 and 125 µg/mL (C18MImCl) for ATCC30010 and isolate T4, respectively. C16MImNTf2 did not demonstrate amoebicidal activity. All active IS caused the hemolysis of erythrocytes. The cytotoxic effect of the IS was tested in RAW macrophages and human brain microvascular endothelial cells, which demonstrated cytotoxicity in all concentrations tested against both cell lines. As a consequence, these IS with amoebicidal activity presented low selectivity index values (SI) (SI < 1.0), demonstrating lack of parasite selectivity. CONCLUSION: Thus, C16MImMeS, C16MImCl, and C18MImCl seem to hold greater promise as components for contact lens cleaning and disinfection solutions, instead of direct human application.
Asunto(s)
Acanthamoeba castellanii/efectos de los fármacos , Amebicidas/farmacología , Imidazoles/farmacología , Acanthamoeba castellanii/crecimiento & desarrollo , Amebicidas/química , Animales , Encéfalo/irrigación sanguínea , Encéfalo/citología , Células Cultivadas , Células Endoteliales , Ambiente , Hemólisis , Humanos , Imidazoles/química , Concentración 50 Inhibidora , Cinética , Imagen por Resonancia Magnética , Ratones , Pruebas de Sensibilidad Parasitaria , Células RAW 264.7 , Trofozoítos/efectos de los fármacosRESUMEN
Several strains of free-living amoebae belonging to the genus Acanthamoeba can cause a painful sight-threatening disease of the cornea known as Acanthamoeba keratitis (AK). The numbers of AK cases keep rising worldwide mainly due to an increase in contact lens wearers and lack of hygiene in the maintenance of contact lenses and their cases. We report a case of AK in a healthy young woman admitted to the Hospital de Clinicas in Porto Alegre, southern Brazil. Corneal scrapings were examined for the presence of Acanthamoeba strains. The initial isolate was characterized by morphological and genotypic properties. The isolate belonged to group III according to Pussard and Pons' cyst morphology. Analysis of its 18S rDNA sequence identified the isolate as genotype T4. The T4 genotype is the most commonly reported among keratitis isolates and the most common in environmental samples.
Asunto(s)
Queratitis por Acanthamoeba/parasitología , Acanthamoeba/patogenicidad , Lentes de Contacto/parasitología , Córnea/parasitología , Acanthamoeba/genética , Acanthamoeba/aislamiento & purificación , Acanthamoeba/ultraestructura , Queratitis por Acanthamoeba/epidemiología , Queratitis por Acanthamoeba/etiología , Adulto , Brasil/epidemiología , Córnea/patología , ADN Protozoario/genética , ADN Ribosómico/genética , Femenino , Genotipo , Humanos , Microscopía , Análisis de Secuencia de ADN , NataciónRESUMEN
Free-living amoebae (FLA) are widely distributed in soil and water. A few number of them are implicated in human disease: Acanthamoeba spp., Naegleria fowleri, Balamuthia mandrillaris and Sappinia diploidea. Species of Acanthamoeba can cause keratitis and brain infections. In this study, 72 water samples were taken from both hot tubs and thermal swimming pools in the city of Porto Alegre, RS, Brazil, to determine the presence of Acanthamoeba in the water as well as perform the phenotypic and genotypic characterization of the isolates. The identification of the isolates was based on the cysts morphology and PCR amplification using genus-specific oligonucleotides. When the isolates were submitted to PCR reaction only 8 were confirmed as belonging to the genus Acanthamoeba. The sequences analysis when compared to the sequences in the GenBank, showed genotype distribution in group T3 (12,5%), T5 (12,5%), T4 (25%) and T15 (50%). The results of this study confirmed the presence of potentially pathogenic isolates of free living amoebae in hot swimming pool and spas which can present risks to human health.