Aislamiento de Acanthamoeba spp. en salas cerradas de un hospital de la provincia de Buenos Aires, Argentina / Isolation of Acanthamoeba spp. in closed rooms of a hospital in Buenos Aires province, Argentina

Resumen Las amebas de vida libre (AVL) son protozoos ubicuos con cuatro géneros patógenos para el ser humano: Acanthamoeba, Naegleria, Balamuthia, y Sappinia. Acanthamoeba puede actuar como reservorio de microorganismos (endosimbiontes), por lo cual, en medio hospitalario, implicaría un riesgo para la transmisión de bacterias, virus y hongos intranosocomiales. Se investigó la presencia de AVL, con énfasis en Acanthamoeba spp., en un hospital pediátrico de la provincia de Buenos Aires, Argentina. Se colectaron 22 muestras de lavamanos e incubadoras en salas de Neonatología y Terapia Intensiva, las que fueron cultivadas a 37 y 42 °C. Los aislados fueron identificados molecularmente. El 63,64% de las muestras presentaron Acanthamoeba spp. Esta investigación representa el primer estudio realizado en la Argentina sobre la detección de Acanthamoeba spp. en salas cerradas de un hospital. Su presencia es una señal de alarma y resulta un blanco útil para investigar posibles reservorios de microorganismos patógenos en ambientes hospitalarios.

Abstract Free-living amoebae (FLA) are ubiquitous protozoa with four pathogenic genera for humans: Acanthamoeba, Naegleria, Balamuthia, and Sappinia. Acanthamoeba can act as a reservoir of microorganisms (endosymbionts), for which reason, in a hospital environment, it would imply a risk for transmission of nosocomial bacteria, viruses and fungi. The presence of AVL, with emphasis on Acanthamoeba spp., was investigated in a pediatric hospital. Twenty-two samples were collected from sinks and incubators in Neonatology and Intensive Care rooms, which were cultured at 37 and 42 °C. The isolates found were molecularly identified. A total of 63.64% of the samples presented Acanthamoeba spp. This research represents the first study in Argentina on the detection of Acanthamoeba spp. in closed rooms of a hospital. Its presence is an alarm signal, and it is a useful target to investigate possible reservoirs of pathogenic microorganisms in hospital environments.

Resumo As amebas de vida livre (AVL) são protozoários ubíquos com quatro gêneros patogênicos para o ser humano: Acanthamoeba, Naegleria, Balamuthia, e Sappinia. Acanthamoeba pode atuar como um reservatório de microrganismos (endossimbiontes), e portanto, em um ambiente hospitalar, representaria um risco de transmissão de bactérias, vírus e fungos intra-nosocomiais. A presença de AVL, com em Acanthamoeba spp. em um hospital pediátrico da província de Buenos Aires, Argentina, foi investigada. Vinte e duas amostras foram coletadas em lavatórios e incubadoras em Salas de Neonatologia e Cuidados Intensivos, cultivadas a 37 e 42 °C. Os isolados foram identificadas molecularmente. Foram encontradas Acanthamoeba spp. em 63,64% das amostras. Esta investigação representa o primeiro estudo realizado na Argentina sobre a detecção de Acanthamoeba spp. em salas fechadas de um hospital. A sua presença é um sinal de alarme e um alvo para investigar possíveis reservatórios de microrganismos patogênicos em ambientes hospitalares.

Presencia de Acanthamoeba spp. en piscinas recreativas al aire libre de la ciudad de Córdoba, Argentina / Presence of Acanthamoeba spp. in outdoor recreational swimming pools in Córdoba city, Argentina / Presença de Acanthamoeba spp. em piscinas recreativas ao ar livre da cidade de Córdoba, Argentina

Resumen El objetivo del estudio fue determinar la presencia de Acanthamoeba spp. en piscinas de uso recreacional, al aire libre, de la ciudad de Córdoba, Argentina. Se recolectaron 30 muestras de agua correspondientes a un total de 10 piscinas. Estas se sembraron en agar no nutritivo en presencia de Escherichia coli en solución de Page. Luego de 72 horas de incubación a 37 °C, la identificación genérica se realizó mediante criterios morfológicos. La prueba de transformación amebo-flagelar se realizó para diferenciar amebas de vida libre que desarrollan trofozoítos flagelados, como Naegleria. Se midieron parámetros fisicoquímicos en cada una de las piscinas. Se identificó Acanthamoeba spp. en el 20% del total de las muestras. Las piscinas en las que se aisló este parásito presentaron niveles no detectables de cloro residual, pH moderadamente alcalino y temperatura templada. Este estudio demuestra la presencia de Acanthamoeba spp. en piscinas recreativas de Córdoba, lo que puede representar un potencial riesgo para la salud pública.

Abstract The aim of the study was to determine the presence of Acanthamoeba spp. in outdoor recreational pools, in Córdoba city, Argentina. Thirty water samples corresponding to a total of 10 pools were collected. These samples were sown on a non-nutritive agar in the presence of Escherichia coli in Pages's solution. After 72 hours of incubation at 37 °C, the generic identification was done based on morphological criteria. The amoebo-flagellate transformation test was performed to differentiate from genera that develop flagellated trophozoites, such as Naegleria. Physicochemical parameters were measured in each of the pools. Acanthamoeba spp. was identified in 20% of the samples. The pools, where this parasite was isolated, presented undetectable levels of residual chlorine, moderately alkaline pH and warm temperature. This study demonstrates the presence of Acanthamoeba spp. in recreational pools in Córdoba, which may represent a potential risk to public health.

Resumo O objetivo do estudo foi determinar a presença de Acanthamoeba spp. em piscinas para uso recreativo, ao ar livre, na cidade de Córdoba, Argentina. Foram coletadas 30 amostras de água correspondentes a um total de 10 piscinas. Elas foram semeadas em ágar não nutritivo na presença de Escherichia coli em solução de Page. Após 72 horas de incubação a 37 °C, a identificação genérica foi realizada utilizando critérios morfológicos. O teste de transformação amebo-flagelar foi realizado para diferenciar amebas de vida livre que desenvolvem trofozoítos flagelados, como Naegleria. Parâmetros físico-químicos foram medidos em cada uma das piscinas. Acanthamoeba spp. foi identificada em 20% do total das amostras. As piscinas onde este parasita foi isolado apresentaram níveis indetectáveis de cloro residual, pH moderadamente alcalino e temperatura temperada. Esse estudo demonstra a presença de Acanthamoeba spp. em piscinas recreativas de Córdoba, o que pode representar um risco potencial para a saúde pública.

Encefalitis amebiana granulomatosa por Amebas de Vida Libre en un paciente pediátrico / Granulomatous amoebic encephalitis due to free-living amebas in a pediatric patient

Introducción: Las amebas de vida libre (AVL) Naegleria fowleri, Acanthamoeba spp., Balamuthia mandrillaris, son protozoos ampliamente distribuidos en la naturaleza. Son microorganismos oportunistas, que afectan preferentemente al sistema nervioso central causando cuadros de meningoencefalitis amebiana primaria o de encefalitis granulomatosa amebiana (EGA), ambas patologías de alta mortalidad. Descripsión del Caso: Paciente femenina de 10 años, que ingresa a nuestro hospital por presentar hemiparesia braquiocrural izquierda, vómitos y cefalea de 72 horas de evolución. Se realiza tomografía computada y Resonancia Magnética en la que se identifica una lesión expansiva, sólido-quística ubicada en la región fronto derecha. Se realiza exéresis completa de la misma y se implementa tratamiento farmacológico lográndose la restitución ad-integrum de la función motora y la resolución de la EGA a 30 meses de seguimiento. Discusión: La encefalitis granulomatosa amebiana es una patología causada por amebas de vida libre. Su presentacíon clínica puede ser indistinguible de otras causas de leptomeningitis o encefalitis, como las de origen bacteriano, viral o por micobacterias lo que impide conocer su real incidencia. Su tratamiento antiparasitario es muy complejo y su evolución es habitualmente fatal. Conclusión: Reportamos un caso de Encefalitis Amebiana Granulomatosa en una paciente pediátrica inmunocompetente, con excelente evolución.

Introduction: Free-living amoebas (FLA) Naegleria fowleri, Acanthamoeba spp., and Balamuthia mandrillaris, are protist widely distributed in nature. Are opportunistic microorganisms, preferentially affect the central nervous system causing primary amoebic meningoencephalitis or amoebic granulomatous encephalitis (AGE), both with high mortality. Case report: A 10 year-old female patient was admitted with a three-day history of left hemiparesis accompanied with headaches and vomiting. Computed tomography and magnetic resonance were performed, in which an expansive solid cystic mass was observed in the right fronto-parietal region. Complete resection was performed and pharmacological treatment was started, achieving complete restitution of motor function and resolution of AGE after 30 months of follow-up. Discussion: AGE is a desease caused by free-living amoebas. Its clinical presentation is similar to other leptomeningitis or encephalitis of different etiology such as bacterial, viral or by mycobacterial, which cannot know its real incidence. Its pharmacological treatment is complex and its evolution is usually fatal. Conclusion: We report a case of Amoebic Granulomatous Encephalitis in an immunocompetent pediatric patient with good outcome.

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)

Balamuthia mandrillaris en el Perú, lesiones cutáneas, meningoencefalitis y métodos de cultivo / Balamuthia mandrillaris in Perú, skin lesions, meningoencephalitis and cultivation methods

Las amebas de vida libre son microorganismos abundantes en el suelo y agua en todo el planeta. Algunas especies de estas amebas son capaces de causar muerte en el ser humano y animales, asi como Naegleria fowleri (N. fowleri), Acanthamoeba sp. T4 principalmente y Balamuthia mandrillaris (B. mandrillaris). El 98% de los casos reportados en el mundo han fallecido. En Peru alrededor de 8 casos de meningoencefalitis por Acanthamoeba sp. se han reportado y mas de 55 casos por B. mandrillaris . Ningun caso por N. fowleri ha sido documentado oficialmente en el Peru. B. mandrillaris es de dificil diagnostico y aislamiento en medios de cultivo. En esta revision pretendo documentar la historia de casos reportados de los ultimos 40 años y describir los medios de cultivo utiles para su crecimiento. La identificacion morfologica y molecular de Balamuthia es critica en el diagnostico de meningoencefalitis amebiana.

Free-living amoebae are abundant microorganisms in soil and water worldwide. Some species of these amoebae are capable of causing death in humans and animals, such as Naegleria fowleri (N. fowleri) , Acanthamoeba sp . T4 , and Balamuthia mandrillaris (B. mandrillaris) . Some 98% of cases reported in the world have resulted in death. In Peru, 8 cases of meningoencephalitis due to Acanthamoeba sp. have been reported and more than 55 cases per B. mandrillaris . No case of N. fowleri has been officially documented in Peru. B. mandrillaris is difficult to diagnose and isolate in culture media. In this review we document the history of reported cases of the last 40 years and describe useful methods for their growth. Morphological and molecular identification of Balamuthia is critical to the diagnosis of amoebic meningoencephalitis.

Reporte de caso y revisión de la literatura de caso de paciente con meningoencefalitis por amebas de vida libre / Case report and literature review of a patient with meningoencephalitis from free-living amoebae

Paciente de 14 años, proveniente de Apartadó (Antioquia), remitido al Hospital Pablo Tobón Uribe de Medellín por un cuadro febril agudo asociado a deterioro rápido del estado de conciencia, decantándose posteriormente como causa de su deterioro una ameba de vida libre (Naegleria fowleri) . Las amebas de vida libre son una causa etiológica poco común de meningitis, tienen un curso fulminante y requieren de un alto índice de sospecha para hacer un tratamiento oportuno.

A 14-year-old patient from Apartado (Antioquia, Colombia) was referred to Hospital Pablo Tobon Uribe in Medellin for acute febrile illness associated with headache and rapid loss of consciousness. The etiologic agent causing this deterioration was found to be a free-living amoeba ( Naegleria fowleri ). Free-living amoebae are a rare etiology for meningoencephalitis, have a fulminant course and require a high index of suspicion for early diagnosis.

Acanthamoeba spp. en piscinas cubiertas de la ciudad de Bahía Blanca, provincia de Buenos Aires, Argentina / Acanthamoeba spp. in indoor swimming pools in Bahía Blanca city, Buenos Aires province, Argentina

Acanthamoeba ha sido aislada de numerosos hábitats incluyendo piletas de natación. Estudiar su distribución es importante ya que algunas especies causan enfermedad en el hombre. El objetivo del presente trabajo fue la búsqueda, aislamiento y caracterización de protozoos del género Acanthamoeba en piscinas cubiertas de la ciudad de Bahía Blanca, Argentina, en las cuatro estaciones del año durante 2007-2008. Se estudiaron 7 piscinas y en cada una se tomaron cuatro muestras: fondo, superficie, raspado de pared y para análisis bacteriológico. Las muestras se analizaron por observación directa y por cultivo en agar no nutritivo a 37 °C y a 42 °C. La identificación genérica se realizó de acuerdo con las características morfológicas de quistes y trofozoítos y para identificar Naegleria se realizó la prueba de transformación ameboflagelar. En 5 de las 7 piscinas se aislaron amebas de vida libre al menos en una época del año. La prueba de transformación ameboflagelar resultó negativa, descartando al género Naegleria. Todos los aislamientos correspondieron al género Acanthamoeba Grupos II y III de Pussard y Pons. Si bien la eliminación de Acanthamoeba en las aguas de natatorios resulta muy difícil por tratarse de un protozoario ubicuo y sumamente resistente a los desinfectantes comúnmente utilizados, se recomienda una limpieza profunda de las piscinas que minimice los riesgos de infección.

Acanthamoeba spp. has been isolated from many habitats, including swimming pools. Investigations on its distribution are relevant because many of its species cause human diseases. The aim of the present work was to investigate, isolate and characterize protozoan of the genus Acanthamoeba from indoor swimming pools in Bahía Blanca, Argentina, in the four seasons, during the 2007-2008 period. Seven pools were studied and samples were collected from the bottom, surface and wall by scraping them. Besides, samples for bacteriological analysis were taken from each pool. The samples were analyzed by direct observation and by culturing on non nutritive agar at 37 °C and at 42 °C. The generic identification was performed according to the morphologic characteristics of cysts and trofozoites, while the amoebo-flagellate transformation test was carried out to identify Naegleria. Free-living amoebas were isolated from five of the seven swimming pools examinated, in at least one season. Naegleria genus was not found as the amoebo-flagellate transformation tests were negative in all samples. All the isolations corresponded to the genus Acanthamoeba belonging to Pussard & Pons morphological Groups II and III. Although elimination of Acanthamoeba from the water is difficult because it is a ubiquitous protozoan. which is highly resistant to the commonly used disinfectants, a thorough cleaning of the pools to minimize the risks of infection is recommended.

Free-living Acanthamoeba and Naegleria spp. amebae in water sources of León, Nicaragua

free-living amebae (FLA) are known to occur worldwide in water-related biotopes, but only limited information is available on these organisms in developing countries and so far no information on their presence is available from Nicaragua. The aims of this study were to evaluate the prevalence of potentially pathogenic Acanthamoeba spp. and Naegleria spp. in different water sources to which the population of León municipality is exposed. Since pathogenic amebae are thermotolerant, we were especially interested in the occurrence of FLA in geothermal areas. Water samples were collected from León area in Nicaragua: 88 samples were from rivers and springs, 111 from wells, 74 from water taps and 21 from water tanks in urban and suburban León and from three nearby geothermal areas of San Jacinto, Posoltega and Tipitapa. Amebae were identified using morphological and physiological criteria, immunohistochemical staining procedures and molecular methods. Indirect immunofluorescent test was performed on cysts and trophozoites fixed on microscopical slides and incubated for 30 min at room temperature in separate experiments with the following antibodies: rabbit-anti N. fowleri/N. lovanensis (Nf-Pab), mouse monoclonal antibody anti N. fowleri (Nf-5D12u), rabbit antibodies against Acanthamoeba spp. And fluorescent in situ hybridization (FISH) was performed using 18S rRNA-targeted fluorescent oligonucleotide probes. Probes: GSP for the detection of Acanthamoeba and NAEG1088 for the detection of Naegleria. Free-living amebae were recovered from approximately 43 % of the samples. Acanthamoeba spp was found in 21 % of samples from León municipality and in 2 % of samples from geothermal areas. Amoeboflagellates were found in 10 % of samples from León and in 19 % in geothermal areas. Fifty three percent of tested wells in the geothermal area contained thermotolerant amoeboflagellates. Naegleria spp. was identified in 24 out of 39 (61.5 %) of isolated amoeboflagellates. Twelve of them were assigned to N. lovanenesis while none of the isolates could be identified as N. fowleri. However, the common presence of thermotolerant Naegleria in water, specially N. lovanensis, which is an indicator species for N. fowleri, suggests that also this pathogenic amoeba may pose a risk to public health in the area. On the other side, direct pathogenicity, free-living amebae are receiving increasing attention as reservoirs and potential vehicles for the transmission of bacteria in the environment. Thus the information provided in this study may serve as base-line for future studies on the role of free-living amebae e.g. in waterborne-disease outbreaks in the region. Among such potentially important enteropathgens are Vibrio cholerae, E. coli 0157, and Helicobacter pylori. Rev. Biol. Trop. 56 (2): 439-446. Epub 2008 June 30.

Las amebas de vida libre (AVL) son un grupo de organismos de distribución mundial. Entre las AVL hay parásitos facultativos en humanos y otros animales, los cuales pertenecen a los géneros Acanthamoeba, Naegleria y Balamuthia que causan infecciones severas en el sistema nervioso central. Sin embargo no se tiene ninguna información de Nicaragua. El objetivo de este estudio fue evaluar la presencia de amebas de vida libre, en diferentes fuentes de agua de la parte urbana y rural del Departamento de León, y áreas geotérmicas de Nicaragua. Estas amebas fueron identificadas usando criterios morfológicos, fisiológicos, histoquímicos y moleculares. En los resultados se encontró amebas de vida libre en el 43% del total de las muestras. En la municipalidad de León, se encontraron 21% de Acanthamopeba sp. y en las áreas geotérmicas un 2%. Las amebaflagelados tipo Naegleria fueron 10% y 19% respectivamente. Del grupo amebaflagelados fueron 24, de las cuales 12 se dentificaron como N. lovanensis. En estas muestras no se aisló N. fowleri (ameba patógena), sin embargo, la presencia de N. lovanensis es in indicador de la presencia de N. fowleri, la cual puede ser un riesgo a la salud pública. Además, estas amebas pueden servir como vectores de bacterias enteropatógenas.