Acanthamoeba spp. y Naegleria spp. aisladas del Río Pamplonita, zona metropolitana de Cúcuta, Colombia / Acanthamoeba spp. and Naegleria spp. isolated from Pamplonita river, metropolitan area of Cucuta, Colombia

Introducción: Acanthamoeba y Naegleria son géneros de amebas de vida libre resistentes a cambios extremos de temperatura y pH, aislados de diversos ambientes (suelo, aire y agua). Debido a la cantidad de habitantes que se benefician de las aguas del Río Pamplonita, al desconocimiento de su presencia en la región y al aumento de enfermedades relacionadas con el consumo de agua, se considera necesario realizar estudios sobre la distribución de estos microorganismos. Objetivo: Identificar Acanthamoeba spp. y Naegleria spp. en aguas del curso principal del río Pamplonita de la zona metropolitana de Cúcuta, Colombia. Métodos: La búsqueda de las amebas de interés se realizó mediante examen directo y cultivo a 28, 37 y 42 °C en agar no nutritivo a partir de 21 muestras de agua de siete sectores del río Pamplonita. Adicionalmente, se midió pH y temperatura in situ y se determinó la carga de coliformes. Resultados: Se encontró que 76,2 por ciento de las muestras fueron positivas para alguna de las amebas. La Acanthamoeba fue la que se aisló con mayor frecuencia. El 28,6 por ciento de los cultivos incubados a 42 °C fueron positivos principalmente para Naegleria spp., lo que indica que estos aislados termotolerantes podrían tener potencial patógeno. Conclusiones: Acanthamoeba spp. y Naegleria spp. son frecuentes en aguas del Río Pamplonita en áreas de importante intervención humana. El hallazgo de amebas termotolerantes alerta sobre el riesgo de salud para la población que se encuentra expuesta a esta fuente hídrica(AU)

Introduction: Acanthamoeba and Naegleria are genus of free-living amoeba (AVL) resistant to extreme changes in temperature and pH, isolated from different environments (soil, air and water). Due to the number of inhabitants that benefit from the waters of Pamplonita River, the lack of knowledge about its presence in the region and the increase of diseases related to water consumption, it is necessary to carry out studies on its distribution. Objective: To identify Acanthamoeba spp and Naegleria spp in the water of the main course of Pamplonita River, in the metropolitan area of Cúcuta, Colombia. Methods: The search for the amoebas of interest was performed by direct examination and culture at 28, 37 and 42 °C on non-nutritive agar from 21 water samples from seven sectors of Pamplonita River. Additionally, pH and temperature were measured in situ and the coliforms´ load was also determined. Results: It was found that 76.2 percent of the samples were positive for some of the amoebas, with Acanthamoeba being the most frequently isolated. 28.6 percent of the cultures incubated at 42 °C were positive mainly for Naegleria spp, indicating that these thermotolerant isolates could have pathogen potential. Conclusions: Acanthamoeba spp and Naegleria spp are frequent in waters of Pamplonita River which are areas of important human intervention. The finding of thermotolerant amoebas warns about the health risk for the population that is exposed to this water source(AU)

Experimental demonstration of the possible role of Acanthamoeba polyphaga in the infection and disease progression in Buruli Ulcer (BU) using ICR mice.

The transmission of Buruli ulcer (BU), caused by Mycobacterium ulcerans (MU), remains puzzling although a number of hypothesis including through bites of infected aquatic insects have been proposed. We report the results of experiments using ICR mice that give credence to our hypothesis that Acanthamoeba species may play a role in BU transmission. We cocultured MU N2 and MU 1615 which expresses red fluorescent protein (RFP) and Acanthamoeba polyphaga (AP), and confirmed infected AP by Ziehl-Neelsen (ZN) staining. We tested for viability of MU inside AP and observed strong RFP signals inside both trophozoites and cysts after 3 and 42 days of coculturing respectively. ICR mice were topically treated, either on shaved intact or shaved pinpricked rumps, with one of the following; MU N2 only (2.25 x 106 colony forming units [CFU] / ml), MU N2:AP coculture (2.96 x 104 CFU: 1.6 x 106 cells/ml), AP only (1.6 x 106 cells/ml), PYG medium and sterile distilled water. Both MU N2 only and MU N2:AP elicited reddening on day (D) 31; edema on D 45 and D 44 respectively, and ulcers on D 49 at pinpricked sites only. To ascertain infectivity and pathogenicity of MU N2 only and MU N2:AP, and compare their virulence, the standard mouse footpad inoculation method was used. MU N2:AP elicited reddening in footpads by D 3 compared to D 14 with MU N2 only of the same dose of MU N2 (2.96 x 104 CFU). ZN-stained MU were observed in both thin sectioned and homogenized lesions, and aspirates from infected sites. Viable MU N2 were recovered from cultures of the homogenates and aspirates. This study demonstrates in ICR mice MU transmission via passive infection, and shows that punctures in the skin are prerequisite for infection, and that coculturing of MU with AP enhances pathogenesis.

Azole Antifungal Agents To Treat the Human Pathogens Acanthamoeba castellanii and Acanthamoeba polyphaga through Inhibition of Sterol 14α-Demethylase (CYP51).

In this study, we investigate the amebicidal activities of the pharmaceutical triazole CYP51 inhibitors fluconazole, itraconazole, and voriconazole against Acanthamoeba castellanii and Acanthamoeba polyphaga and assess their potential as therapeutic agents against Acanthamoeba infections in humans. Amebicidal activities of the triazoles were assessed by in vitro minimum inhibition concentration (MIC) determinations using trophozoites of A. castellanii and A. polyphaga. In addition, triazole effectiveness was assessed by ligand binding studies and inhibition of CYP51 activity of purified A. castellanii CYP51 (AcCYP51) that was heterologously expressed in Escherichia coli. Itraconazole and voriconazole bound tightly to AcCYP51 (dissociation constant [Kd] of 10 and 13 nM), whereas fluconazole bound weakly (Kd of 2,137 nM). Both itraconazole and voriconazole were confirmed to be strong inhibitors of AcCYP51 activity (50% inhibitory concentrations [IC50] of 0.23 and 0.39 µM), whereas inhibition by fluconazole was weak (IC50, 30 µM). However, itraconazole was 8- to 16-fold less effective (MIC, 16 mg/liter) at inhibiting A. polyphaga and A. castellanii cell proliferation than voriconazole (MIC, 1 to 2 mg/liter), while fluconazole did not inhibit Acanthamoeba cell division (MIC, >64 mg/liter) in vitro. Voriconazole was an effective inhibitor of trophozoite proliferation for A. castellanii and A. polyphaga; therefore, it should be evaluated in trials versus itraconazole for controlling Acanthamoeba infections.

Iodamoeba butschlii in an anal pap test confirmed by iodine stain.

We report the finding of Iodamoeba butschlii amebic cysts on a liquid-based anal Pap smear from an HIV-positive male. Iodine staining of the smear confirmed the diagnosis. It is important to distinguish I. butschlii from pathogenic ameobae and other organisms seen on anal Pap smears.

Nuevos métodos para el diagnóstico microbiológico de la amebiasis y de otros protozoarios intestinales / New methods for the microbiological diagnosis of amebiasis and other intestinal protozoa

La Organización Mundial de la Salud (OMS) ubica a las enfermedades diarreicas en segundo lugar como causa de morbi-mortalidad de niños en países en vías de desarrollo, siendo las infecciones por protozoarios intestinales proporcionalmente una causa trascendente en dichas regiones. La mayoría de estos parásitos se transmiten por vía fecal-oral o por contacto interpersonal y exhiben ciclosde vida que, en general se desarrollan en dos estadios: el trofozoíto que coloniza el intestino y el quiste que es excretado con las heces y cuya rígida pared protectora, le confiere resistencia en el ambiente, permitiendo de este modo la propagación de la enfermedad. Entamoeba histolytica es uno de los agentes infecciosos de mayor distribución mundial, y es aproximadamente la causa de 100.000 muertes por año, siendo responsable de uno de los problemas de salud más serios en países en vías de desarrollo. Se reconocen al menos ocho amebas que podrían colonizar el intestino del hombre: E. histolytica, E. dispar, E. moshkovskii, E. coli, E. hartmanni, E. polecki, Iodamoeba butschlii y Endolimax nana. Aunque sólo E. histolytica es considerada como el agente etiológico de la Amebiosis. Su presentación clínica va desde la colonización asintomática, la disentería amebiana, hasta la propagación extraintestinal, originando abscesos en diversos órganos y tejidos. Por este motivo, la correcta identificación de E. histolytica en heces y tejidos y diferenciándola de otras amebas comensales y de otros protozoarios representa un desafío en la práctica médica, a que de ello dependerá el tratamiento y el pronóstico de laenfermedad.

Abstract: The World Health Organization (WHO) includes diarrhea a the second leading cause of morbidity and mortality of children in developing countries. Among these causes, infections by intestinal protozoan parasites represent an important percentage in any place of the world. Most of these parasites are transmitted by the fecal-oral route or by inter personal contact and exhibit simple life cycles, consisting in the disease-causing, proliferating trophozoites and the dormant, resistant cyst responsible for the transmission of the infection among susceptible hosts. Entamoeba histolytica is one of the most frequent of those parasites, causing about 100.000 deaths per year in developing countries, being one of the major health problems in areas where basic sanitation practices are inefficient. At least eight species of Entamoeba have been reported to infect the human large intestine: E. histolytica, E. dispar, E. moshkovskii, E. coli, E. hartmanni, E. polecki, odamoeba butschlii and Endolimax nana, although E. histolytica is the only one considered to cause pathology in humans. Clinical manifestation of this infection varies from asymptomatic infection to dysentery and extraintestinal invasion, producing abscesses in many tissues. Therefore, the correct identification of E. histolytica in stool and tissue samples and its differential diagnostic is an important challenge in parasitology because treatment and prognosis depend of the valid identification of this parasite.The health issue of protozoan intestinal infections, both in developed and developing parts of the world, is of such importance that it is clearly necessary the development of novel, better, cheap, and faster diagnostic methods. The incorporation of new approaches and technology to the efficient and sensitive detection of these infections is significantly relevant.

In the case of transmission of Mycobacterium ulcerans in buruli ulcer disease Acanthamoeba species stand accused.

Buruli ulcer disease caused by Mycobacterium ulcerans results in extensive destruction of skin and soft tissue and long-term functional disabilities that ultimately require surgery and rehabilitation. The disease is associated with aquatic and swampy environments with the mycobacterium occurring in biofilms, soil, aquatic insects, fish and wildlife however, the mode of transmission to humans remains an enigma. Current transmission ideas including bites from predatory water bugs and mosquitoes, do not explain satisfactorily the spasmodic disease distribution in human populations. Here we argue that Acanthamoeba species are the natural hosts of M. ulcerans and are mainly responsible for disease transmission because; (i) Acanthamoebae are known natural hosts of several microbial pathogens including M. marinum, M. avium and Legionella pneumophila, (ii) culture of slow-to-grow microbial pathogens hosted in nature by Acanthamoeba spp is enhanced when the media is seeded with the protozoa, (iii) acanthamoebae and M. ulcerans share similar bio-ecological and epidemiological settings, (iv) documented evidence that prior growth of L. pneumophila and M. avium in acanthamoebae influences entry mechanisms, intracellular growth and virulence in human monocytes, (v) Acanthamoeba spp also infect humans and cause diseases via routes of openings including broken skin and sites of trauma similar to M. ulcerans and (vi) M. ulcerans is rather a fastidious intracellular organism as recent analysis of the genome indicate. We argue further that temperature plays a significant role in transmission determining the fate of either the intracellular microbe or the host cells. Also, Acanthamoeba-pathogen association has a long evolutionary history because the same set of bacterial genes and gene products e.g. in L. pneumophila are required for survival in both mammalian and protozoan host cells. We suggest that the involvement of Acanthamoeba in the transmission of M. ulcerans to humans better explains the disease's epidemiology.

Epizootic fatal amebiasis in an outdoor group of Old World monkeys.

BACKGROUND: Entamoeba (E.) histolytica is an obligate parasite of humans and non-human primates. METHODS: This report describes the pathomorphological, immunohistological, and microbiological findings of fatal E. histolytica infection in two mantled guerezas (Colobus guereza) and one Hanuman langur (Semnopithecus entellus) from an epizootic outbreak of amebiasis in an open-range recreation park. RESULTS: Pathomorphological examination revealed multifocal necrotizing and granulomatous hepatitis with intralesional protozoan trophozoites in all three cases. In addition, necrotizing and ulcerative gastritis was detected in both mantled guerezas. Furthermore, oligofocal acute pulmonary embolization was detected in one of these cases. No extra-hepatic lesions were observed in the Hanuman langur. Immunohistological examination confirmed the etiologic diagnosis of E. histolytica-induced lesions. CONCLUSIONS: Although E. histolytica is a rarely diagnosed pathogen in Western European countries, veterinarians and animal keepers involved in handling and care taking of non-human primates should be aware of the potential threat caused by this zoonotic parasite.

Emerging diseases in tropical dermatology.

All dermatologists worldwide should have at least passing familiarity with various tropical maladies that generate cutaneous manifestions. In addition to the standard infectious ailments associated with tropical environs, the authors have described herein five "emerging" illnesses that are gaining increasing attention for their capacity to cause human disease in those immigrating from, or traveling to, the tropical and subtropical world.

Histopathologic spectrum and immunohistochemical diagnosis of amebic meningoencephalitis.

Traditionally, Naegleria fowleri infections are labeled primary amebic meningoencephalitis because of prominent meningeal neutrophilic inflammation. Acanthamoeba spp. and Balamuthia mandrillaris are labeled granulomatous amebic encephalitis because of parenchymal granulomatous inflammation. We compared histopathologic and immunohistochemical features of 18 cases with central nervous system free-living ameba infections. Immunohistochemical assays using polyclonal antibodies that reacted specifically against each genus identified 11 patients with Balamuthia infection, four with N. fowleri, and three with Acanthamoeba. Immunohistochemical assays highlighted the presence of trophozoites that were difficult to identify with hematoxylin and eosin stains in areas of necrosis or where macrophages were abundant. Immunohistochemical assays also demonstrated the presence of granular antigens inside macrophages and blood vessel walls. Amebic cysts were observed in three patients with Acanthamoeba infection and in three with Balamuthia. Patients with Acanthamoeba infection showed granulomatous inflammation. Patients with Naegleria infection had neutrophilic inflammation. Balamuthia infections showed a spectrum of inflammation that ranged from primarily neutrophils to granulomas. Meningitis was present in 88% of cases. Immunohistochemical assays were useful to demonstrate the presence of granular antigens and confirmed the genus of the ameba. The spectrum of inflammation in cases of Balamuthia meningoencephalitis is broader than previously described. The term amebic meningoencephalitis describes better the histopathologic findings than the currently used classification of primary amebic meningoencephalitis and granulomatous amebic encephalitis.

Microfauna associated with amoebic gill disease in sea-farmed Atlantic salmon, Salmo salar L., smolts.

A study of microfauna, associated with pathological changes in the gills of Atlantic salmon, Salmo salar L., was conducted over 2001-2002. Monthly samples of 1(+) salmon smolts were taken, protozoan populations were quantified and gill health was assessed histologically. Protozoan densities were correlated with pathological changes, in order to determine their possible role in lesions in the gills. The most severe gill tissue changes were observed in summer/autumn and the least in spring. A diverse polyphyletic protozoan community was observed colonizing the gills, including Neoparamoeba sp., other amoebae, scuticociliates, Ichthyobodo-like flagellates, trichodinid ciliates and prostomatean ciliates. The earlier gill tissue changes in the gill were not always associated with the presence of these microorganisms, whereas amoebae (other than Neoparamoeba sp.), Ichthyobodo-like flagellates and trichodinid ciliates correlated with augmenting gill lesions. Neoparamoeba sp. was present, but its abundance did not correlate with the disease. This study suggests that a diversity of protozoans including Ichthyobodo-like flagellates, trichodinid ciliates and amoebae other than Neoparamoeba sp. are involved in the aetiology of amoebic gill disease in the Irish situation.

Ameba-associated microorganisms and diagnosis of nosocomial pneumonia.

To elucidate the role of ameba-associated microorganisms (AAMs) as etiologic agents of pneumonia, we screened for Legionella spp., Parachlamydia acanthamoeba, Afipia sp., Bosea spp., Bradyrhizobium spp., Mesorhizobium amorphae, Rasbo bacterium, Azorhizobium caulinodans, Acanthamoeba polyphaga mimivirus, and conventional microorganisms in 210 pneumonia patients in intensive-care units by using culture, polymerase chain reaction, and serologic testing. These resulted in 59 diagnoses in 40 patients. AAMs and non-AAMs were implicated in 10.5% of the patients. The infectious agents were identified in 15 patients: Acanthamoeba polyphaga mimivirus, 8; Legionella pneumophila, 3; L. anisa, 1; Parachlamydia sp., 1; Bosea massiliensis, L. worsleiensis, L. quinlivanii, and L. rubrilucens, 1; and M. amorphae and R. bacterium, 1. A. polyphaga mimivirus was the fourth most common etiologic agent, with a higher seroprevalence than noted in healthy controls. This finding suggested its clinical relevance. Therefore, AAM might cause nosocomial pneumonia and should be suspected when conventional microbiologic results are negative.

September 2004: a 6-year-old girl with headache and stiff neck.

Free-living amebas in the genera Naegleria, Acanthamoeba and Balamuthia are known to cause CNS infections. Here we report a case of fatal granulomatous amebic meningoencephalitis (GAE) caused by Balamuthia mandrillaris in a 6-year-old previously healthy girl who presented with headache and stiff neck. She was treated medically for brain abscess after a CT scan identified a ring-enhancing lesion in the right temporo-parietal area. A brain biopsy showed necrosis and granulomatous inflammation. Subsequently, multiple new lesions appeared in the brain bilaterally. A second brain biopsy revealed viable amebic trophozoites that were most abundant in perivascular spaces, accompanied by neutrophils, macrophages and eosinophils. Immunofluorescence study confirmed the amoeba as Balamuthia mandrillaris. This case demonstrates that making diagnosis of GAE pre-mortem requires a high index of suspicion. Amebic infection should be included in the differential diagnosis of any granulomatous lesion in CNS; and careful search for amebic parasites should be carried out especially when necrosis predominates in the pathological material.

Isolation of Acanthamoeba sp. from conjunctival sac of healthy individuals using swab.

Is Acanthamoeba sp. normally found in the eyes? A study was carried out to establish the possibility of Acanthamoeba sp. as a part of the normal conjunctival flora. Conjunctiva swabbing were carried out in 286 healthy Orang Asli school children using sterile cotton swab. The swab was then inoculated onto non-nutrient agar (NN-A). Heat killed Escherichia coli that was used as food source for the growth of the amoebae was pipetted onto and away from the smear. The plates were incubated at 30 degrees C and examined daily using an inverted microscope for 14 days. Morphology of the trophozoites and cysts of the amoebae were used as the taxonomic criteria for identification. Positive-controls and negative-controls were done to check for the consistency of the technique used and monitoring of contamination respectively. None of the conjunctiva swab cultured was positive for Acanthamoeba sp. This finding may indicate that Acanthamoeba sp. is not part of normal conjunctival flora or conjunctiva swab is an insensitive technique to isolate the organism. However, a more extensive research is needed to investigate these possibilities.

Pathogenesis of Acanthamoeba infections.

Acanthamoeba are free-living, harmless organisms, however, given the opportunity and the appropriate conditions, they can cause painful, sight-threatening as well as fatal infections and, thus, are considered opportunistic pathogens. Acanthamoeba infections have become increasingly important in the past few years due to increasing populations of contact lens users and AIDS patients. The mechanisms associated with the pathogenesis of Acanthamoeba tend to be highly complex, depending on parasite, host and the environmental factors. Elucidation of the biochemical, cellular and molecular basis of the pathogenesis of diseases caused by Acanthamoeba may lead to the development of therapeutic interventions.

Defense mechanisms of IFN-gamma and LPS-primed murine microglia against Acanthamoeba castellanii infection.

In the central nervous system (CNS), cytokine-primed microglia play a central role in host's defense against Acanthamoeba castellanii infection. In this study, the effect of recombinant interferon (rIFN)-gamma and Salmonella enterica serovar enteritidis lipopolysaccharide (LPS), both inflammatory stimuli, on A. castellanii infection in murine microglia was examined. Priming of microglia with rIFN-gamma and LPS synergistically triggered, in a dose-dependent manner, amebastatic activity in these cells. More than 52%, 88% or 95% of this function was then abrogated by anti-IL-1beta (but not anti-IL-1alpha), IL-6 or TNF-alpha neutralizing antibodies, suggesting that these endogenously produced cytokines may participate in the antimicrobial capacity. Consistent with these findings, the priming of microglia with rIFN-gamma and LPS elicited the release of proinflammatory interleukin (IL)-1alpha, IL-1beta, IL-6 and tumor necrosis factor (TNF)-alpha. Since L-canavanine affected amebastatic activity only during the priming process but not during the infection process, NO-dependent pathway appears to be not the sole antiparasitic mechanism involved in this function. These data suggest that rIFN-gamma and LPS, likely through a proinflammatory network, up-regulate the release of IL-beta, IL-6 and TNF-alpha, which could trigger antimicrobial activity against A. castellanii infection in the brain.

Endogenous amoebic endophthalmitis.

A 42-year-old man experienced abrupt reduction of vision in the left eye and had vitreous opacity and hypopyon on initial examination. He underwent an emergency vitrectomy and phacoemulsification with posterior capsulectomy, followed by a 2-week course of intravenous fluconazole and antibiotics. The final visual acuity was 20/20 OS. Amoebas, together with polymorphonuclear cells, were observed on examination of specimens of the aqueous humor and vitreous fluid obtained during the surgery. Staphylococcus epidermidis was also cultured from the vitreous fluid. The amoebas had a crescent-shaped periphery of the cytoplasm and ambiguous nuclei. Ultrastructurally, the cytoplasm was surrounded by a thick electron-dense coating. The early vitrectomy and combined use of antibiotics and antifungals might have led to the good visual outcome in this patient.

Molecular biology of Entamoeba histolytica: a review

Amebiasis is one of the main causes worldwide of morbidity and mortality by parasites. Application of recombinant DNA technology to the study of Entamoeba histolytica is bringing new light into our understanding of this remarkable protozoan parasite and of the disease it causes. New achievements affect the way we approach many essential questions about E. histolytica, form the mechanism of its pathogenicity to the definition of E. hitolytica as a separate species from the nonpathogenic E. dispar. To give a single example, transfection of trophozoites is now possible and a new generation of studies taking advantage of this capability of manipulation is expected in the short term. Our goal with this review is to provide an updated and simple guide to the growing information on the molecular biology of E. histolytica

Amibiasis espectro clínico y tratamiento

Se denomina amibiasis la infección por Entamoeba histolytica, independientemente de que dé lugar o no manifestaciones clínicas. El que estas se presnten está supeditado a la interaccción entre el parásito, el huésped y diversas peculiaridades del medio ambiente, ya que dependiendo de la correlación de fuerzas entre los agentes que intervienen en dicha interacción, una infección amibiana puede permanecer limitada a la luz del intestino (amibiasis no invasora) o transformarse a corto o largo plazo en una infección que invade la pared y a su vez se constituye en el punto de partida para el desarrollo de diferentes cuadros clínicos. Precisamente a la descripción, diagnóstico clínico, diagnóstico paraclínico y diagnóstico por imágenes, al igual que al tratamiento de cada uno de los diversos síndromes y complicaciones derivadas de la infección por Entamoeba histolytica está dedicada la presente actualización

Contribuiçäo ao estudo da suscetibilidade de Acanthamoeba spp a diferentes métodos de desinfecçäo de lentes de contacto gelatinosas

Desde 1980 vários casos de ceratite por Acanthamoeba têm sido diagnóstico e, ao contrário dos primeiros relatos onde o fator desencadeante era o trauma, atualmente a quase totalidade dos casos relaciona-se com o uso de lentes de contacto. Estudos têm sido desenvolvidos para esclarecer o envolvimento destas lentes na fisiopatologia da ceratite por Acanthamoeba. Dentre estes, a avaliaçäo dos diferentes sistemas de desinfecçäo empregados na sua manutençäo, aponta como um assunto contoverso e de interesse cientifico, uma vez que, um grande número de espécies e cepas estäo envolvidas, apresentando, cada uma delas, um comportamento diferente, em relaçäo a estes sistemas. Este estudo teve por objetivo testar a susceptibilidade de cinco diferentes espécies de Acanthomoeba, A. Astronyxis, A. Polyphaga, A. castellani, A. Cellbertsoni e Acanthamoeba sp a um sistema de desinfecçäo térmica química (Hidrocare e Flex-care). Dois experimentos foram realizados. No primeiro, lentes de contato gelatinosas foram contaminadas com as diferentes espécies do protozoário e em seguida submetidas a desinfecçäo e cultivo. No segundo, cistos e trofozoítos das mesmas espécies foram expostos diretamente aos sistemas de desinfecçäo, sendo posteriormente lavados, com o objetivo de eliminar a soluçäo química eventualmente presentes na superfície da ameba. O controle foi defínido pelo aparecimento de forma trofozoíticas. A desinfecçäo térmica mostrou-se efetiva em 100% dos testes realizados com as cinco diferentes espécies de Acanthamoeba avaliadas. A culbertsoni resistiu a desinfecçäo química com Flexcare, sendo susceptível ao Hidrocare, e Acanthomoeba sp resistiu aos dois compostos testados. O estudo permitiu concluir que, dos métodos testados, a desinfecçäo térmica é a única totalmente efetiva para a desinfecçäo de lentes de contacto gelatinosas contaminadas com Acanthamoeba