Parasitosis ocular por Acanthamoeba / Acanthamoeba parasitic eye infection

RESUMEN El parasitismo es uno de los fenómenos más sorprendentes de los observados en los animales. El número de organismos conocidos de vida parasitaria es muy elevado. Existe una gran cantidad de especies cuya supervivencia está relacionada con la de otras y dependen, en distinta medida, de ellas. Esta dependencia no implica que los animales parásitos sean organismos degenerados o deficientes; al contrario, a la vida parasitaria se ha llegado tras largo tiempo de evolución, en el que los parásitos han ido superando barreras y adaptándose a vivir en, o sobre sus hospedadores. Las queratitis por Acanthamoeba son infrecuentes; pero se describe un aumento a nivel mundial relacionado con el uso creciente de lentes de contacto. El diagnóstico precoz y el tratamiento adecuado deben realizarse para evitar la pérdida de la visión. Se realizó una búsqueda de artículos publicados, con el objetivo de conocer sobre la Acanthamoeba como parásito y su afectación ocular. Se utilizó la plataforma Infomed, específicamente la Biblioteca Virtual de Salud(AU)

ABSTRACT Parasitism is one of the most surprising phenomena among those occurring in animals. The number of known parasitic organisms is very high. There is a large number of species whose survival is related to that of others and depend on them to a greater or lesser degree. Such dependence does not imply that parasitic animals are either degenerate or deficient. On the contrary, parasitic life has been the result of a long process of evolution along which parasites have gradually overcome hurdles and have adapted to live in or on their hosts. Acanthamoeba keratitis is a rare disease, but a worldwide increase has been reported due to the growing use of contact lenses. Early diagnosis and appropriate treatment are required to prevent sight loss. A search was conducted for published papers with the purpose of learning about Acanthamoeba as a parasite and the related eye conditions. Use was made of the platform Infomed, specifically the Virtual Health Library(AU)

Evaluation of the sensitivity to chlorhexidine, voriconazole and itraconazole of T4 genotype Acanthamoeba isolated from Mexico.

Free-living amoebae of the genus Acanthamoeba are the etiological agents of cutaneous lesions, granulomatous amoebic encephalitis (GAE) and amoebic keratitis (AK), which are chronic infections with poor prognosis if not diagnosed promptly. Currently, there is no optimal therapeutic scheme to eradicate the pathologies these protozoa cause. In this study we report the morphological and molecular identification of three species of the genus Acanthamoeba, belonging to T4 group; A. polyphaga isolated from the corneal ulcer of a patient sample of AK case; A. castellanii isolated from the contact lens of an AK patient and A. palestinensis obtained from a soil sample. The in vitro activity of chlorhexidine, itraconazole and voriconazole drugs against trophic stage was also evaluated through a colorimetric assay based on the oxidation-reduction of alamar blue. The strains in the study were sensitive to the evaluated drugs; although when determining the 50% inhibitory concentration (IC50) statistically significant differences were observed. A. castellanii showed to be highly sensitive to voriconazole (0.66 ±â€¯0.13 µM) but the least sensitive to chlorhexidine and itraconazole (8.61 ±â€¯1.63 and 20.14 ±â€¯4.93 µM, respectively), A. palestinensis showed the highest sensitivity to itraconazole (0.502 ±â€¯0.11 µM) and A. polyphaga expressed moderate sensitivity to chlorhexidine and itraconazole and lower sensitivity to voriconazole (10.10 ±â€¯2.21 µM). These results showed that species of the genus Acanthamoeba express different sensitivity to the tested drugs, which could explain the problems surrounding the establishment of a treatment of choice in the infections caused by these amoebae. We consider that although chlorhexidine and itraconazole show good activity on these amoebae and have been used in cases of AK in Mexico with acceptable results, voriconazole should be considered as the first therapeutic option of future Acanthamoeba infections that will be diagnosed in our country.

Acanthamoeba keratitis in Mexico: Report of a clinical case and importance of sensitivity assays for a better outcome.

Acanthamoeba keratitis (AK) is a sight-threatening corneal infection. The early symptoms include redness, pain, photophobia and intense tearing. Chronic infection usually progresses to stromal inflammation, ring ulcers, corneal opacification and hypopyon. Here we document an AK case in a high myopic 38-year-old woman from Mexico City, with a history of wearing contact lenses while swimming. Corneal scrapes cultures were positive only for amoebae, consequently a treatment including netilmicin 0.3% and oral itraconazole 100 mg/12 h was prescribed. The infection was resolved after 8 months, leaving a slight leucoma outside the visual axis, with a visual acuity of 20/150. In the laboratory, the amoebic isolate was axenized in PYG medium, with an optimal growth at 30 °C, and was identified morphologically as Acanthamoeba polyphaga according to the taxonomic criteria of Page (1988) and placed in the T4 group by genotyping. The virulence of this strain (40%) was determined by intranasal inoculation of 1 × 106/20 µl trophozoites in BALB/c mice recovering from brain, proving their invasion ability and by the interaction with monolayers of epithelial cells of the established MDCK line of canine kidney origin (1:2 ratio of interaction), at 1, 3, 6, 8 and 24 h; trophozoites migrated to cell junctions inducing few lytic zones. In addition to the biological characterization, in vitro drug sensitivity tests were performed using chlorhexidine, itraconazole, netilmicin and voriconazole. Results revealed that voriconazole was the most effective compound. A. polyphaga remains as one of the most frequently isolated species producing AK. The treatment of AK case using netilmicin and oral itraconazole solved the disease, but the healing process was wide-ranging (8 months). The use of voriconazole and chlorhexidine may be an alternative treatment of future AK cases in Mexico.

The first Acanthamoeba keratitis case in the Midwest region of Brazil: diagnosis, genotyping of the parasite and disease outcome.

We report an Acanthamoeba keratitis case associated with the use of contact lens in a 28-year-old female from Brasília, Brazil. Samples from corneal scraping and contact lens case were used for culture establishment, PCR amplification, and partial sequencing (fragments of ~400kb) of small subunit rDNA; both culture and PCR were positive. The sequence analyses of the cornea and of isolates from the contact lens case showed similarity with the T4 genotype. To the best of our knowledge, this is the first report of T4 Acanthamoeba keratitis case from the Midwest region of Brazil.

The first Acanthamoeba keratitis case in the Midwest region of Brazil: diagnosis, genotyping of the parasite and disease outcome

Abstract We report an Acanthamoeba keratitis case associated with the use of contact lens in a 28-year-old female from Brasília, Brazil. Samples from corneal scraping and contact lens case were used for culture establishment, PCR amplification, and partial sequencing (fragments of ~400kb) of small subunit rDNA; both culture and PCR were positive. The sequence analyses of the cornea and of isolates from the contact lens case showed similarity with the T4 genotype. To the best of our knowledge, this is the first report of T4 Acanthamoeba keratitis case from the Midwest region of Brazil.

Virulent T4 Acanthamoeba causing keratitis in a patient after swimming while wearing contact lenses in Southern Brazil.

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.

Molecular and morphological characterization of Acanthamoeba isolated from corneal scrapes and contact lens wearers in Argentina.

In this study, we describe the frequency of Acanthamoeba keratitis (AK) in patients that assisted in the Ophthalmology Department and determine the species/genotypes of free living amoebas (FLA) isolates. FLA from Corneal scrapes (CS) and contact lens (CL) wearers were studied by morphological and molecular characterization. A database was constructed with sociodemographic, clinical findings and history of use of CL variables. During January 2000 and September 2016 patients with corneal pathology admitted to the Ophthalmology Service of the University Hospital in Córdoba city, Argentina were included in the study. FLA were detected in 1.5% (11/739) and in 17% (11/65) of CS and CL analyzed respectively. FLA isolates from CL users evidenced an 80.9% of inappropriate lens maintenance, 4.8% (1/21) were not CL users that have been in contact with waters in outdoor environment and 14,3% (3/21) with no data about CL users. Acanthamoeba was confirmed in 100% and 82% of CS and LC respectively. The most frequent symptom associated with AK was red eye and photophobia. FLA from CS belonged to group II but 82% (9/11) and 18% (2/11) from CL belonged to group II and III respectively. T4 genotype and A. polyphaga species were detected in 100% of Acanthamoeba isolates. Poor CL hygiene practices, highlights the need for improved education about the severity of AK and consequences of improper CL hygiene. Genotype T4 detected in 100% of both CS and CL samples, consistently with previous findings indicating that this genotype is by far the most prevalent isolated from ocular infection.

Acanthamoeba genotypes T3 and T4 as causative agents of amoebic keratitis in Mexico.

Free-living amoebae (FLA) are widely distributed worldwide. Some genera included in this group act as opportunistic pathogens causing fatal encephalitis and Acanthamoeba keratitis (AK), a sight-threatening infection of the cornea associated with the use of soft contact lenses that could even end in blindness if an early diagnosis and treatment are not achieved. Furthermore, the numbers of AK cases keep rising worldwide mainly due to an increase of contact lens wearers and lack of hygiene in the maintenance of lenses and their cases. In Mexico, no cases of AK have been described so far although the isolation of other pathogenic FLA such as Naegleria fowleri and Balamuthia mandrillaris from both clinical and environmental sources has been reported. The present study reports two cases of Acanthamoeba keratitis diagnosed in two patients admitted to the Hospital "Luis Sánchez Bulnes" for Blindness Prevention in Mexico City, Mexico. Corneal scrapes and contact lenses were checked for the presence of Acanthamoeba strains in both patients. Strains were axenized after initial isolation to classify at the genotype level. After sequencing the diagnostic fragment 3 (DF3) region located on the 18S ribosomal DNA (rDNA) gene of Acanthamoeba, genotype T3 and genotype T4 were identified in clinical case 1 and 2, respectively. To our knowledge, these are the first reported cases of AK in Mexico in the literature and the first description of Acanthamoeba genotypes T3 and T4 as causative agents of amoebic infection.

Acanthamoeba spp: ecoepidemiología, biología, ultraestructura, patogénesis y diagnóstico en el hombre / Acanthamoeba: ecoepidemiology, biology, ultrastructure, pathogenesis and diagnosis in humans

En el presente trabajo presentamos nuestros estudios referidos al protozoo Acanthamoeba, desde lo morfológico, diagnóstico de laboratorio, patología en el ser humano, aspectos epidemiológicos y resistencia al cloro. Se aisló Acanthamoeba en el 28.6% de los tanques domiciliarios de agua, en el 71% de piscinas cubiertas, en la totalidad de las muestras del arroyo Napostá, que cruza la ciudad de Bahía Blanca, y no fue detectada en el agua de la red. Con referencia a la acción quisticida del cloro, se logró inhibir el crecimiento recién a las tres horas de exposición en 60 ppm de cloro. Realizamos el primer aislamiento de un paciente con queralitis, en Bahía Blanca. Hasta el momento no aislamos Acanthamoeba de otras muestras biológicas. Con referencia a los aspectos morfológicos, documentamos la ultraestructura de las cepas regionales de Acanthamoeba y realizamos la tipificación molecular de la cepa aislada del paciente con queralitis, una afección que, si bien es de baja prevalencia y su diagnóstico etiológico es de fácil ejecución y bajo costo, no justifica no realizar el estudio oportuna y rápidamente. La sensibilidad de la observación directa por microscopia óptica fue del 11.43%, comparada con los cultivos. Finalmente efectuamos un listado de recomendaciones para usuarios de lentes de contacto.(AU)

Acanthamoeba spp: ecoepidemiología, biología, ultraestructura, patogénesis y diagnóstico en el hombre / Acanthamoeba: ecoepidemiology, biology, ultrastructure, pathogenesis and diagnosis in humans

En el presente trabajo presentamos nuestros estudios referidos al protozoo Acanthamoeba, desde lo morfológico, diagnóstico de laboratorio, patología en el ser humano, aspectos epidemiológicos y resistencia al cloro. Se aisló Acanthamoeba en el 28.6% de los tanques domiciliarios de agua, en el 71% de piscinas cubiertas, en la totalidad de las muestras del arroyo Napostá, que cruza la ciudad de Bahía Blanca, y no fue detectada en el agua de la red. Con referencia a la acción quisticida del cloro, se logró inhibir el crecimiento recién a las tres horas de exposición en 60 ppm de cloro. Realizamos el primer aislamiento de un paciente con queralitis, en Bahía Blanca. Hasta el momento no aislamos Acanthamoeba de otras muestras biológicas. Con referencia a los aspectos morfológicos, documentamos la ultraestructura de las cepas regionales de Acanthamoeba y realizamos la tipificación molecular de la cepa aislada del paciente con queralitis, una afección que, si bien es de baja prevalencia y su diagnóstico etiológico es de fácil ejecución y bajo costo, no justifica no realizar el estudio oportuna y rápidamente. La sensibilidad de la observación directa por microscopia óptica fue del 11.43%, comparada con los cultivos. Finalmente efectuamos un listado de recomendaciones para usuarios de lentes de contacto.

Acanthamoeba castellanii: morphological analysis of the interaction with human cornea.

The present study demonstrates that when Acanthamoeba castellanii trophozoites are co-cultivated with isolated human corneas, the amoeba can be invasive and cause damage to the intact corneal epithelium without the requirement of previous corneal abrasion. After adhesion, A. castellanii trophozoites migrate between cells forming bumps on the corneal cell layers and reaching Bowman s membrane in 3h, although no evidence of cell damage was observed until the phagocytic process was detected. Likewise, conditioned medium produced damage to the corneal cells that was proportional to the time of incubation, but this cytophatic effect involved only the most superficial layer of the human cornea and was not enough to explain amoebic invasion of Bowman s membrane. As a result of our observations, we suggest that the mechanical action of the trophozoites and phagocytosis of corneal cells during the process of corneal invasion are more important than previously suggested.

Acanthamoeba spp. in domestic tap water in houses of contact lens wearers in the metropolitan area of Mexico City.

A survey was carried out in the metropolitan area of Mexico City to determine the presence of Acanthamoeba in the tap water of houses of contact lens wearers. Water samples were taken from the mains water entry, bathroom sinks and storage containers (roof tanks, cisterns) of 27 houses; and from the solution contained in the contact lens cases. Samples were filtered and cultured onto NNE medium. The isolates were identified based on their morphological features and pathogenicity. Total and fecal coliforms, water temperature, pH, dissolved oxygen and residual free-chlorine were measured by standard methods. Forty five isolates of Acanthamoeba from 200 water samples were obtained. The highest number of amoebae was isolated from cisterns and roof tanks. Most Acanthamoeba isolates were non-pathogenic, however, their presence in tap water is a potential hazard since some species can cause Acanthamoeba keratitis and granulomatous amoebic encephalitis.

Examen histopatológico a lente de contacto paciente con queratitis por Acanthamoeba: informe preliminar / Histopathologic examination to contact lens in patient with keratitis by Acanthamoeba: preliminary report

Un grupo de amebas de vida libre (AVL) es capaz de producir afecciones graves en la especie humana, tales como Meningoencefalitis Amebiana Primaria; Encefalitis Granulomatosa Amebiana; y Queratitis por colonización del epitelio corneal en individuos inmunocompetentes, especialmente en usuarios permanentes de lentes de contacto. Se procede a efectuar estudio histológico a lente de contacto proveniente de paciente de 45 años, con diagnóstico presuntivo de queratitis por Acanthamoeba sp. Y cuya muestra de raspado de córnea resulto positivo en medio de cultivo ANNE. Las muestras fueron teñidas con PAS y observadas al microscopio con 4x, 10x y 40x, observándose estructuras altamente sugerentes de quistes de este grupo de amebas. Por la importancia que requiere obtener un diagnóstico rápido y certero en esta patología ocular, debido a que los métodos tradicionales demoran alrededor de una semana, consideramos relevante dar a conocer este hallazgo citohistológico.

Human IgA inhibits adherence of Acanthamoeba polyphaga to epithelial cells and contact lenses.

Specific anti-Acanthamoeba IgA antibodies have been detected in the serum and tears of patients and healthy individuals. However, the role of human secretory IgA antibodies in inhibiting the adherence of Acanthamoeba had not been previously investigated. Therefore, the purpose of this study was to purify secretory IgA from human colostrum and analyze its effect on the adherence of Acanthamoeba trophozoites to contact lenses and Madin-Darby canine kidney (MDCK) cells. IgA antibodies to Acanthamoeba polyphaga in colostrum of healthy women as well as in saliva and serum of healthy subjects were analyzed by ELISA and Western blot analysis. In serum, saliva, and colostrum, we detected IgA antibodies that recognized several antigens of A. polyphaga. In addition, colostrum and IgA antibodies purified from it inhibited adherence of A. polyphaga trophozoites to contact lenses and MDCK cells. These results suggest that IgA antibodies may participate in the resistance to the amoebic infection, probably by inhibiting the adherence of the trophozoites to contact lenses and corneal epithelial cells.