ABSTRACT
RESUMEN Dientamoeba fragilis es un protozoário que parasita el intestino grueso del hombre y animales domésticos. Hasta el momento, aún no son claros aspectos de su ciclo de vida, como el rango de hospedadores, reservorios, mecanismo de infección, entre otros. Se postula que el cerdo es un hospedador natural para este protozoario y que, debido a su cercanía con el humano, podría facilitar una transmisión zoonótica. En Colombia no existen a la fecha estudios sobre la presencia de D. fragilis en hospedadores animales, incluidos los cerdos, y los datos sobre la prevalencia de otros protozoarios intestinales en estos animales son escasos. El objetivo fue determinar la frecuencia de protozoarios intestinales, incluyendo D. fragilis, en cerdos de una granja tecnificada de una zona rural de Medellín (región Andina de Colombia). Se recolectaron muestras de materia fecal de 70 cerdos en etapa de preceba, que fueron evaluadas mediante examen directo, tinción tricrómica y Ziehl-Neelsen modificado. Se realizaron análisis univariados con medidas de frecuencia relativa y tendencia central y análisis bivariados para la exploración de factores de riesgo. Se detectó la presencia de D. fragilis en un 13% de las muestras y de otros parásitos como Entamoeba spp. (66%), Blastocystis spp. (64%), Balantioides coli (36%), Cryptosporidium spp. (36%), Iodamoeba butschlii (17%), coccidias (4%) y Giardia spp. (1,4%). En este primer reporte de D. fragilis en cerdos en Colombia se observó la alta prevalencia de otros protozoarios patógenos, lo que corrobora el papel de los cerdos como importantes reservorios de infecciones humanas. Estudios sobre la presencia de este protozoario tanto en cerdos como en el personal encargado de su manejo contribuirían al conocimiento sobre su dinámica de transmisión.
ABSTRACT Dientamoeba fragilis is a protozoan that parasitizes the large intestine of humans and domestic animals. To date, some aspects regarding D. fragilis life cycle, including hosts, reservoirs, infection mechanism, among others, are not yet clear. Swine are considered natural hosts for this protozoan, therefore their close contact with humans promotes its zoonotic transmission. In Colombia there are no studies on the presence of D. fragilis in animal hosts, including pigs, and data about other intestinal protozoa are scarce. The objective was to determine the prevalence of intestinal protozoan, including D. fragilis, in pigs raised in a farm from a rural area of Medellin (Andean region of Colombia). Fecal samples from 70 pigs in prefattening stage were collected. Direct fecal smear examination, trichrome and modified Ziehl-Neelsen stain were used in the study. Univariate analysis (frequency distribution and central tendency measures) and bivariate analysis were used to explore risk factors. Dientamoeba fragilis was found in 13% of the evaluated fecal samples. Other parasites detected included: Entamoeba spp. (66%), Blastocystis spp. (64%), Balantioides coli (36%), Cryptosporidium spp. (36%), Iodamoeba butschlii (17%), coccidias (4%), and Giardia spp. (1,4%). This is the first report of D. fragilis in swine in Colombia, and the high prevalence of other pathogen protozoa was also observed, which corroborates the role of pigs as important reservoirs for human infections. Studies focused on the evaluation of both swine and swine-exposed farm workers should be done in order to know the dynamics of transmission of this parasite.
Subject(s)
Animals , Parasites , Protozoan Infections, Animal , Swine , Disease Reservoirs , Zoonoses , Risk Factors , Dientamoeba , Intestine, Large , Animals, Domestic , Water Reservoirs , Entamoeba , InfectionsABSTRACT
BACKGROUND: Because of a lack of quality control (QC) materials, stool examination has not been standardised. This study examined intestinal parasites in diarrhea specimens to manufacture and evaluate the performance stability of QC materials for stool examination. METHODS: This study examined diarrhea specimens submitted for stool culture. Microscopic examination was performed using the direct smear and formalin-ether concentration method (Military General Laboratory, MGL). Enzyme-linked immunosorbent assay (ELISA) kits (R-Biopharm AG, Germany) and xTAG Gastrointestinal Pathogen Panel (Luminex Corp., USA) were used for the three major protozoa: Cryptosporidium parvum, Giardia lamblia, and Entamoeba histolytica. Polymerase chain reaction (PCR) was performed for Dientamoeba fragilis and Blastocystis hominis. The QC materials for stool examination were generated using Diphyllobothrium nihonkaiense ova. The manufactured QC materials were evaluated under different storage conditions, with varying preservatives, temperatures, and storage times. RESULTS: From November 2015 to April 2016, 82 diarrhea specimens were collected and tested. All results from microscopy and ELISA were negative; C. parvum (n=2) and G. lamblia (n=1) were detected by xTAG, while D. fragilis (n=10) and B. hominis (n=2) were detected by PCR. High- and low-concentration QC materials were manufactured. Using the high-concentration QC material, ova were observed in all storage conditions using MGL. Using the low-concentration QC material, the ova were observed until 14 days, but not after 3 weeks. CONCLUSIONS: It should be considered for making QC materials for stool examinations that focus on D. fragilis and B. hominis frequently found in Korea and with the caution to the low-concentration of QC materials could be unstable.
Subject(s)
Blastocystis hominis , Cryptosporidium parvum , Diarrhea , Dientamoeba , Diphyllobothrium , Entamoeba histolytica , Enzyme-Linked Immunosorbent Assay , Giardia , Giardia lamblia , Korea , Methods , Microscopy , Ovum , Parasites , Polymerase Chain Reaction , Quality ControlABSTRACT
El análisis de las parasitosis a diferentes escalas (regional, local) se facilita por el Geographic Information System (GIS), que permite identificar áreas con distinto grado de vulnerabilidad. El objetivo del trabajo fue analizar la distribución espacial de las parasitosis intestinales en el partido de La Plata, estableciendo su relación con las condiciones socio-ambientales para identificar áreas con diferente riesgo epidemiológico. Se completó una encuesta epidemiológica y se tomaron muestras seriadas de materia fecal, analizadas por la técnica de Ritchie. Se calcularon niveles de precariedad y vulnerabilidad (Iv), y se compararon con los resultados parasitológicos. Se analizaron parasitológica y ambientalmente 653 personas, de las cuales 585 (89.6%) eran niños y 68 (10.4%) adultos. El análisis indicó que los más vulnerables (Iv = 3-4) estaban en el sector con menor acceso a servicios de infraestructura urbana. El índice se encontró fuertemente asociado a la prevalencia total de parasitosis intestinales y a las 4 especies patógenas asociadas a la contaminación fecal del ambiente: Trichuris trichiura, Ascaris lumbricoides, Hymenolepis nana y Giardia lamblia (p < 0.01). Enterobius vermicularis, parásito no vinculado a la condición sanitaria de sus hospedadores, no se asoció al Iv. Hubo asociación estadística entre precariedad y parasitosis y se halló que a mayor Iv mayor frecuencia de parasitosis (p < 0.01). El uso de GIS permitió zonificar las variables socioambientales en un gradiente creciente de condiciones desfavorables y su relación con la presencia de especies patógenas. La continuidad de estos estudios en diferentes regiones de Argentina contribuye a la determinación de áreas en riesgo sanitario.
Parasitosis analysis at regional levels is simplified by the use of the Geographic Information System (GIS), which enables the identification of areas with different degrees of vulnerability. We analyzed the spatial distribution of intestinal parasites in La Plata district and their relationship with socio-environmental conditions in order to identify areas with different degrees of epidemiological risk. An epidemiological survey was completed; stool samples were collected and analyzed by Ritchie's method. Levels of precariousness and vulnerability (Iv rate) were calculated and compared to the parasitological results. Parasitological and environmental analysis were carried out on a total of 653 individuals, of whom 585 (89.6%) were children and 68 (10.4%) adults. The analysis indicated that the most vulnerable (Iv = 3-4) were those located in areas with less access to urban infrastructure services. The Iv was strongly associated with the overall prevalence of intestinal parasitosis and the 4 pathogenic species related to the fecal contamination of the environment: Trichuris trichiura, Ascaris lumbricoides, Hymenolepis nana and Giardia lamblia (p < 0.01). On the other hand, Enterobius vermicularis, a parasite not related to the sanitary condition of its host, was not associated with the Iv. Statistical association between poverty and parasitosis was noted; the greater the Iv, higher the incidence of parasites (p < 0.01). GIS allowed zoning socio-environmental variables in an increasing gradient of unfavorable conditions and their relationship to the presence of pathogenic species. The continuity of these studies in different regions of Argentina contributes to the determination of health risk areas.
Subject(s)
Adult , Animals , Child , Female , Humans , Male , Intestinal Diseases, Parasitic/epidemiology , Intestines/parasitology , Amoebida/isolation & purification , Argentina/epidemiology , Dientamoeba/isolation & purification , Feces/parasitology , Geographic Information Systems , Giardia lamblia , Helminths/isolation & purification , Incidence , Prevalence , Risk Assessment , Retortamonadidae/isolation & purification , Socioeconomic Factors , Sanitation/statistics & numerical data , Urban PopulationABSTRACT
INTRODUCTION: Studies strongly indicate Dientamoeba fragilis as one of the causes of diarrhea in human immunodeficiency virus (HIV) patients. METHODS: The objective of the present study was to evaluate the prevalence of D. fragilis associated with the causes of diarrhea in 82 HIV/ AIDS patients hospitalized at the Instituto de Infectologia Emílio Ribas from September 2006 to November 2008. RESULTS: In total, 105 samples were collected from 82 patients. Unprotected sex was the most frequent cause of HIV infection (46.3%), followed by the use of injectable or non-injectable drugs (14.6%). Patients presented with viral loads of 49-750,000 copies/ mL (average: 73,849 ± 124,850 copies/mL) and CD4 counts ranging of 2-1,306 cells/mm³ (average: 159 ± 250 cells/mm³). On an average, the odds of obtaining a positive result by using the other techniques (Hoffman, Pons and Janer or Lutz; Ritchie) were 2.7 times higher than the chance of obtaining a positive result by using the simplified iron hematoxylin method. Significant differences were found between the methods (p = 0.003). CONCLUSIONS: The other techniques can detect a significantly greater amount of parasites than the simplified iron hematoxylin method, especially with respect to Isospora belli, Cryptosporidium sp., Schistosoma mansoni, and Strongyloides stercoralis, which were not detected using hematoxylin. Endolimax nana and D. fragilis were detected more frequently on using hematoxylin, and the only parasite not found by the other methods was D. fragilis.
INTRODUÇÃO: Estudos indicam a Dientamoeba fragilis como uma das causas de diarréia em pacientes com HIV/AIDS. MÉTODOS: Os objetivos deste estudo foram avaliar a prevalência de D. fragilis associadas com as causas de diarréia em pacientes com HIV/AIDS internados no Instituto de Infectologia Emílio Ribas (IIER). Oitenta e dois pacientes internados no IIER fizeram parte deste estudo de setembro de 2006 a novembro de 2008. RESULTADOS: No total, 105 amostras foram coletadas a partir de 82 pacientes neste estudo. Sexo desprotegido foi à causa mais frequente para a aquisição do HIV (46,3%), seguido pelo uso de drogas injetáveis ou não injetáveis (14,6%). Relações heterossexuais foram os mais citados (19,5%). Pacientes apresentaram carga viral entre 49 e 750.000 (média de 7.849 ± 124.850) e CD4 variando de 2 a 1.306 (média de 159 ± 250). Em média, as chances de um resultado ser positivo com outras técnicas foram 2,7 vezes maiores do que a chance de um resultado positivo com hematoxilina férrica simplificada. Foram encontradas diferenças significativas entre os métodos (p=0,003). CONCLUSÕES: As outras técnicas são capazes de detectar uma quantidade significativa maior de parasitas em comparação com a hematoxilina férrica simplificada, especialmente em relação à Isospora belli, Cryptosporidium sp., Schistossoma mansoni e Strongyloides stercoralis que não foram encontrados utilizando a hematoxilina e a Endolimax nana e D. fragilis foram mais detectados pela hematoxilina férrica simplificada, principalmente a D. fragilis que não foi detectada pelos outros métodos.
Subject(s)
Adult , Female , Humans , Male , Diarrhea/parasitology , Dientamoeba/isolation & purification , Dientamoebiasis/diagnosis , Feces/parasitology , HIV Enteropathy/parasitology , HIV Enteropathy/diagnosis , Hematoxylin , Prevalence , Staining and LabelingABSTRACT
The diagnosis of D. fragilis by microscopic identification of the parasite in stool is time consuming and relatively insensitive. To evaluate microscopy, culture and PCR for detection of D. fragilis in stool samples and to identify the genotypes of D. fragilis isolates among the study population. A total of 82 fresh human fecal samples were examined by microscopy using Wheatley's trichrome permanent staining, culture using the modified Boeck and Drbohlav medium and PCR targeting the small subunit [SSU] rRNA gene. Additionally, the existence of genetic variation among D. fragilis isolates [proved positive by PCR] was examined by restriction fragment length polymorphism analysis. PCR detected 25 isolates [30.5%], MBD culture detected 24 isolates [29.3%], while microscopy detected 8 isolates [9.8%]. Sensitivities of PCR, culture and microscopy were 92.6%, 88.9% and 29.6%, respectively. The agreement between PCR and culture results was substantial [KA=0.86]. PCR followed by RFLP analysis revealed the existence of two genetic variants among 25 D. fragilis isolates. Genotype I predominated in 23 [92%] samples, while the remaining two isolates corresponded to genotype II. It is recommended to use culture for routine diagnosis of D. fragilis in suspected gastrointestinal cases. Two genetic variants of D. fragilis existed in the Egyptian isolates
Subject(s)
Humans , Male , Female , Dientamoeba/genetics , Genotype , Polymerase Chain Reaction/methods , Microscopy , Culture Media , Comparative StudyABSTRACT
A diarreia, manifestação clínica comum em pacientes acometidos pela AIDS, pode levar a óbito em decorrência do diagnóstico tardio e da falta de tratamento adequado. Uma das causas de diarréia em pacientes com AIDS é atribuída ao Dientamoeba fragilis, cuja etiologia envolve diversos fatores, com a prevalência de patógenos emergentes e oportunistas, como: vírus da imunodeficiência adquirida (HIV) e outros agentes virais, bacterianos e fúngicos. Este estudo avaliou a eficiência da coloração de hematoxilina férrica simplificada na identificação da Dientamoeba fragilis, bem como verificou a prevalência do protozoário associada a causas de diarreia em pacientes com HIV/AIDS. A pesquisa contemplou 82 pacientes com idade compreendida entre 18 e 61 anos (idade média de 38 anos), com predominância do gênero masculino (75,6%), acompanhados no Instituto de Infectologia Emílio Ribas, de setembro de 2006 a novembro de 2008. Das amostras coletadas, com emprego de técnicas especiais de coloração (Hematoxilina Férrica), comparando-se a outros métodos com solução conservante Schaudinn, métodos coproparasitológicos (Hoffman, Pons e Janer, 1934; Richie, 1948). A técnica de hematoxicilina férrica simplificada detectou a Dientamoeba fragilis em 1,2% das amostras, sendo que as outras técnicas não foram capazes de detectar o protozoário. Concluiu-se que a coloração de hematoxilina férrica simplificada mostrou-se eficaz na detecção da Dientamoeba fragilis, é uma técnica rápida e prática para diagnóstico em laboratório de rotina, quando comparada com outros métodos de coloração. A Dientamoeba fragilis foi associada a uma das causas de diarreia em pacientes aidéticos.
Subject(s)
Humans , Male , Female , Acquired Immunodeficiency Syndrome , Diarrhea , Dientamoeba , HematoxylinABSTRACT
A diarreia, manifestação clínica comum em pacientes acometidos pela AIDS, pode levar a óbito em decorrência do diagnóstico tardio e da falta de tratamento adequado. Uma das causas de diarréia em pacientes com AIDS é atribuída ao Dientamoeba fragilis, cuja etiologia envolve diversos fatores, com a prevalência de patógenos emergentes e oportunistas, como: vírus da imunodeficiência adquirida (HIV) e outros agentes virais, bacterianos e fúngicos. Este estudo avaliou a eficiência da coloração de hematoxilina férrica simplificada na identificação da Dientamoeba fragilis, bem como verificou a prevalência do protozoário associada a causas de diarreia em pacientes com HIV/AIDS. A pesquisa contemplou 82 pacientes com idade compreendida entre 18 e 61 anos (idade média de 38 anos), com predominância do gênero masculino (75,6%), acompanhados no Instituto de Infectologia Emílio Ribas, de setembro de 2006 a novembro de 2008. Das amostras coletadas, com emprego de técnicas especiais de coloração (Hematoxilina Férrica), comparando-se a outros métodos com solução conservante Schaudinn, métodos coproparasitológicos (Hoffman, Pons e Janer, 1934; Richie, 1948). A técnica de hematoxicilina férrica simplificada detectou a Dientamoeba fragilis em 1,2% das amostras, sendo que as outras técnicas não foram capazes de detectar o protozoário. Concluiu-se que a coloração de hematoxilina férrica simplificada mostrou-se eficaz na detecção da Dientamoeba fragilis, é uma técnica rápida e prática para diagnóstico em laboratório de rotina, quando comparada com outros métodos de coloração. A Dientamoeba fragilis foi associada a uma das causas de diarreia em pacientes aidéticos.
Subject(s)
Male , Female , Humans , Diarrhea , Dientamoeba , Hematoxylin , Acquired Immunodeficiency SyndromeABSTRACT
It's a retrospective study dealing with 30573 stool exams and 5844 scotch tape tests done in the parasitology-mycology laboratory of Sfax hospital [Tunisia] during 1997 to 2006. 26.6% of stool exams were positive. Third of parasitized patients were children. Protozoa were found in 96.5% Flagellae [54.3%] dominated by Dientamaeba fragilis [30.3%] and Giardia lamblia [17%]; Amoeba [41 .9%] with E histolytica/E dispar in 2.2% of cases. Cryptosporidium parvum was found in 17 cases; Microsporidium sp in 2 HIV positive and Isospora belli in one case. Helminths found in 3.5% were represented by Enterobius vermicularis [49%], Hymenolepis nana [31.4%], Strongyloides stercoralis [0.3%], Taenia saginata [0.3%] and Ankylostoma duodenale [one case]. The social movements occurring in Sfax characterized by rural migration and installation of young persons working in the market gardening could maintain the parasitism frequency
Subject(s)
Humans , Male , Female , Retrospective Studies , Protozoan Infections , Helminths , Flagella , Dientamoeba , Giardia , Amoeba , Entamoeba histolytica , Cryptosporidium parvum , Microsporidia, Unclassified , Ancylostoma , Taenia saginata , Strongyloides stercoralis , Hymenolepis nana , Enterobius , IsosporaABSTRACT
The present study was conducted to determine the prevalence and clinical features of dientamoebiasis in patients presumed to be infected with intestinal parasites. A total of 168 patients were examined for D. fragilis using microscopy [after Wheatley's trichrome staining] and culture [using modified Boeck and Drbohlav's medium] D. fragilis trophozoites were detected in 15 samples [8.9%] examined using trichrome staining and in 50 samples [29.8%] by culture method. Other enteric parasites were common in the study population as 48.8% of patients [82/168] were found harboring intestinal parasites. Blastocystis hominis was the most common, identified in 33.3% [56/168] of the samples. Giardia lamblia was detected in 17.9% [30/168] and E. histolytica/E. dispar in 11.9% [20/168]. The symptoms most frequently encountered were diarrhea, abdominal pain and weight loss and fatigue. Diarrhea and abdominal pain were significantly more frequent in patients with dientamoebiasis compared to non pathogenic cases [P<0.05]. Diarrhea was 38.5% of patients infected with D. fragilis compared to 50% of patients infected with G. lamblia, while abdominal pain was encountered with D. fragilis in 41% compared to 33.3% with G. lamblia. These differences were insignificant [P>0.05]
Subject(s)
Humans , Male , Female , Dientamoeba/parasitology , Prevalence , Feces/parasitology , Signs and Symptoms , Diarrhea , Surveys and QuestionnairesABSTRACT
Se realizó un estudio de purificación del DNA de los trofozoitos de Dientamoeba fragilis a partir de muestras fecales humanas congeladas, no fijadas en formol, y con elevadas concentraciones de inhibidores de la DNA polimerasa. Para tal fin, se utilizaron columnas ("spin-columns") disponibles en el comercio. Esta metodología se comparó con una técnica tradicional de purificación con fenol/cloroformo. Ambos métodos fueron seguidos de la amplificación del DNA de D. fragilis por una "nested" PCR y posterior electroforesis en un gel de agarosa de los amplicones obtenidos. La extracción del DNA a partir de trofozoitos de D. fragilis - mediante el procedimiento de las columnas - produjo DNA de elevada calidad, libre de impurezas e inhibidores de la polimerasa. Por el contrario, con la técnica de fenol/cloroformo se observó la inhibición de la enzima, cuando se analizaron los productos de amplificación. Además, se confirmó que el agregado de SAF (solución de acetato de sodio - ácido acético - formol) a las muestras fecales afecta la integridad del DNA y como consecuencia impide su posterior amplificación.
Subject(s)
Humans , DNA, Protozoan/isolation & purification , DNA-Directed DNA Polymerase , Dientamoeba/genetics , Feces/parasitology , Polymerase Chain Reaction/methods , Culture Media , Electrophoresis, Agar Gel , Molecular Sequence DataABSTRACT
Differential diagnosis of Entamoeba histolytica and Entamoeba dispar, which are two morphologically identical species of amoebae is essential both for the treatment decision and public health aspects. The electrophoretic patterns of hexokinase and phosphoglucomutase have been widely used to distinguish E. histolytica from E. dispar and isoenzyme analysis is still a gold standard method. This study was done to determin the isoenzyme patterns of E. histolytica and E. dispar in Iran. In this study electrophoretic patterns of two Iranian Entamoeba isolates and four standard E. histolytica and E. dispar were analyzed by using four enzymes: Glucose phosphate isomerase [GPI], phosphoglucomutase [PGM], L-malate: NADP[+] oxido-reductase [ME] and hexokinase [HK]. Both Iranian isolates gave zymodeme I pattern of E. dispar. After axenization of the isolates in YIGADHA-S medium, the electerophoretic patterns of zymodeme I remained stable without any changes. Our study confirmed the existence of zymodeme I of E. dispar in Iran. The result of this research and another experiment on 8 isolates led us to declare that zymodeme I might be the dominant pattern of E. dispar zymodemes in Iran
Subject(s)
Humans , Protozoan Proteins , DientamoebaSubject(s)
Dientamoeba/cytology , Dientamoeba/parasitology , Dientamoebiasis/diagnosis , Dientamoebiasis/parasitology , Dientamoebiasis/pathology , Dysentery, Amebic/diagnosis , Dysentery, Amebic/parasitology , Dysentery, Amebic/pathology , Entamoebiasis/diagnosis , Entamoebiasis/parasitology , Entamoebiasis/pathology , Staining and LabelingABSTRACT
Se realizó una experiencia para evaluar la efectividad de las preparaciones permanentes de muestras obtenidas previa purga en fijador de Brooke-Goldman (PVA), con muestras espontáneas de los mismos pacientes, en fijador de Junod (SAF). También fueron comparados los porcentajes obtenidos para E. histolytica y D. fragilis, con los modelos de recolección y procesamiento de heces más usados en nuestro país. Fueron estudiados 248 pacientes. Se tomaron tres muestras de cada uno de ellos. La primera (M1) y segunda muestra (M2) fueron recolectados con heces espontáneas, de días no consecutivos, en fijador de Junod (SAF) y, una tercera (M3), previa purga, en fijador de Brooke-Goldman (PVA). Las muestras recolectadas en SAF (M1 y M2) fueron analizadas por microscopia de preparaciones húmedas directas y previo enriquecimiento por los métodos Ritchie y Faust. Las muestras fijadas en PVA se colorearon con la técnica tricrómica modificada. El análisis de los datos obtenidos mostró diferencias significativas entre las muestras espontáneas de días no consecutivos (M1 y M2) para: Entamoeba histolytica, Giordia lamblia y Endolimax nana. También el examen estadístico reveló diferencias significativas entre las muestras uno y dos, tomadas juntamente, con respecto a los resultados obtenidos en PVA (M3), para: Entamoeba histolytica, Giardia lambia, Dientamoeba fragilis y Endolimax nana. Se compararon los porcentajes obtenidos por coloraciones permanentes para E. hitolytica y D. fragilis, con los correspondientes a otras formas de diagnóstico: 9.088 muestras procesadas por los métodos directo y Ritchie, y 4.228 estudiados por los métodos directo, Ritchie, Faust y coloraciones sólo en casos sospechosos. Estos datos indicaron una mayor eficiencia de la recolección en PVA, previa purga y posterior coloración para el diagnóstico de los dos protozoarios.