RESUMEN
We report a case of Enterocytozoon bieneusi infection in a pediatric hematopoietic stem cell transplant recipient in Argentina. Spores were visualized in feces using Calcofluor White and modified trichrome stainings. PCR and sequencing identified E. bieneusi genotype D in fecal samples and liver samples, confirming extraintestinal dissemination of the parasite.
Asunto(s)
Enterocytozoon , Trasplante de Células Madre Hematopoyéticas , Humanos , Niño , Argentina/epidemiología , Enterocytozoon/genética , Receptores de Trasplantes , Heces , Trasplante de Células Madre Hematopoyéticas/efectos adversosRESUMEN
Several PCR methodologies are available for the detection of Enterocytozoon hepatopenaei (EHP) that target the SSU rRNA gene. However, these methodologies are reported as unsuitable for the detection of EHP due to specificity issues. Here, we report the applicability of two commonly used SSU rRNA methodologies for the detection of additional microsporidia from the genus Vittaforma that is present in cultured Penaeus vannamei from Costa Rica. The molecular detection of DNA of the novel microsporidia can only be achieved using SSU rRNA targeting methodologies and does not cross-react with the highly specific spore wall protein gene PCR detection method.
Asunto(s)
Enterocytozoon , Microsporidia no Clasificados , Microsporidios , Penaeidae , Animales , Microsporidia no Clasificados/genética , Penaeidae/genética , Vittaforma/genética , Costa Rica , Reacción en Cadena de la Polimerasa/métodos , Enterocytozoon/genética , Microsporidios/genética , ARN RibosómicoRESUMEN
The aetiology of diarrhoea in a patient in Cuba with HIV was investigated. Although molecular diagnostics are still not used in many under-resourced settings, here traditional methods were supported by use of PCR. This approach enabled detection of a dual infection (Cystoisospora belli and Enterocytozoon bieneusi), the latter of which was not identified by microscopy with Didier's trichromic staining.
Asunto(s)
Coccidiosis/diagnóstico , Diarrea/diagnóstico , Enterocytozoon/aislamiento & purificación , Microsporidiosis/diagnóstico , Sarcocystidae/aislamiento & purificación , Adulto , Antiinfecciosos/uso terapéutico , Coccidiosis/tratamiento farmacológico , Cuba , Diarrea/microbiología , Diarrea/parasitología , Enterocytozoon/genética , Infecciones por VIH/complicaciones , Infecciones por VIH/tratamiento farmacológico , Humanos , Huésped Inmunocomprometido , Masculino , Microsporidiosis/tratamiento farmacológico , Técnicas de Diagnóstico Molecular , Reacción en Cadena de la Polimerasa , Sarcocystidae/genética , Sarcoma de Kaposi/complicaciones , Sarcoma de Kaposi/tratamiento farmacológico , Combinación Trimetoprim y Sulfametoxazol/uso terapéuticoRESUMEN
Abstract Microsporidia are obligate intracellular fungi with a remarkable ability to infect a wide range of invertebrate and vertebrate hosts. Namely, Enterocytozoon bieneusi is the most frequently microsporidia reported worldwide, and mainly associated with chronic diarrea and wasting syndrome in AIDS patients. Microscopy and PCR-based detection techniques are effective for diagnosis and identification of species and genotypes; however, these methods should be standardized in each laboratory. In this study, we performed microscopy and nested PCR techniques with PCR product sequencing to detect E. bieneusi in human stool samples. These techniques, if applied together, might prove useful for diagnosis and future epidemiological studies of intestinal microsporidiosis in Argentina.
Resumen Los microsporidios son hongos intracelulares obligados con una notable capacidad para infectar una amplia gama de hospedadores invertebrados y vertebrados. Enterocytozoon bieneusi es el microsporidio más frecuentemente reportado en todo el mundo, principalmente tricrómicaasociado con diarrea crónica y síndrome debilitante en pacientes con sida. Las técnicas dedetección basadas en microscopía y PCR son útiles para el diagnóstico y la identificación deespecies y genotipos, pero estos métodos deben estar estandarizados en cada laboratorio.En este estudio evaluamos técnicas de microscopía y PCR anidada, con secuenciación de losproductos, para detectar E. bieneusi en muestras de heces humanas. Estas técnicas, usadas con-juntamente, podrían ser útiles para su aplicación en el diagnóstico de microsporidiosis intestinaly para realizar estudios epidemiológicos de esta afección en Argentina.
Asunto(s)
Humanos , Microsporidios , Enterocytozoon , Esporas Fúngicas , Reacción en Cadena de la Polimerasa , Microsporidios/genética , Enterocytozoon/genética , HecesRESUMEN
Microsporidia are obligate intracellular fungi with a remarkable ability to infect a wide range of invertebrate and vertebrate hosts. Namely, Enterocytozoon bieneusi is the most frequently microsporidia reported worldwide, and mainly associated with chronic diarrhea and wasting syndrome in AIDS patients. Microscopy and PCR-based detection techniques are effective for diagnosis and identification of species and genotypes; however, these methods should be standardized in each laboratory. In this study, we performed microscopy and nested PCR techniques with PCR product sequencing to detect E. bieneusi in human stool samples. These techniques, if applied together, might prove useful for diagnosis and future epidemiological studies of intestinal microsporidiosis in Argentina.
Asunto(s)
Enterocytozoon , Microsporidios , Enterocytozoon/genética , Heces , Humanos , Microsporidios/genética , Reacción en Cadena de la Polimerasa , Esporas FúngicasRESUMEN
BACKGROUND: Enterocytozoon bieneusi is the most common microsporidian species causing diarrhea and other intestinal disorders in humans and animals. Like other infectious diseases, microsporidiosis usually disproportionately affects poor populations. In China, some ethnic minority areas remain poor. Currently, no information of E. bieneusi infection is available in minority populations. The present aims were to understand occurrence and genetic characterizations of E. bieneusi in ethnic minority groups from a poverty-stricken ethnic township in Yunnan Province, and to assess risk factors for E. bieneusi infection. METHODOLOGY/PRINCIPAL FINDINGS: 289 fecal specimens were collected from Yao people (one specimen each) with and without diarrhea, in Yunnan Province. E. bieneusi was identified and genotyped by PCR and sequence analysis of the ITS region of the rRNA gene. An average prevalence of 8.30% (24/289) was observed and four genotypes were identified-genotype Peru6 (n = 21) and three novel genotypes (one each). Genotype Peru6 was detected in two family members in each of three families. In a phylogenetic analysis, all of four genotypes fell into group 1 with zoonotic potential. The people owning individual pit toilets had a statistically higher prevalence of E. bieneusi (16.67%, 12/72) than those using public pit toilets (6.06%, 12/198). CONCLUSIONS/SIGNIFICANCE: This is the first report on occurrence and genetic characteristics of E. bieneusi in ethnic minority groups in China. Genotype Peru6 was found in humans in China for the first time and showed dominance in Yao people. The same genotype was found in some family members and all the genotypes fell into group 1, suggesting the possibility of anthroponotic and zoonotic transmissions. The majority (83.33%, 20/24) of E. bieneusi positive individuals did not present diarrhea. In any case, it is important to recognize their existence and the importance that asymptomatic individuals to E. bieneusi may have from an epidemiological point of view, as transmitters of this pathogen. The analysis of risk factors provides scientific evidence for the development of effective strategies for prevention and control of E. bieneusi infection.
Asunto(s)
Enterocytozoon/genética , Enterocytozoon/aislamiento & purificación , Microsporidiosis/microbiología , Adulto , Anciano , China/epidemiología , China/etnología , Enterocytozoon/clasificación , Etnicidad , Femenino , Variación Genética , Humanos , Masculino , Microsporidiosis/epidemiología , Microsporidiosis/etnología , Persona de Mediana Edad , Grupos Minoritarios , Filogenia , Reacción en Cadena de la Polimerasa , Factores de Riesgo , Adulto JovenRESUMEN
Abstract Enterocytozoon bieneusi is an opportunistic intestinal pathogen that infects humans and a wide variety of animals worldwide. Our aim in this study was to investigate the occurrence of E. bieneusi in a domestic cat population in Campo Grande, Mato Grosso do Sul, Brazil. Sixty fecal samples from diarrheic cats were subjected to polymerase chain reaction (PCR) and the amplicons were sequenced for identification. E. bieneusi was detected in two samples (3.3%), both identified as genotype D. This genotype has already been reported in animals and humans and is considered a zoonotic genotype. Our findings represent the first report of E. bieneusi in domestic cats in Brazil, reinforcing the importance of identifying this agent as a source of infection in animals and humans.
Resumo Enterocytozoon bieneusi é um patógeno intestinal oportunista que infecta humanos e uma variedade de animais em todo o mundo. O objetivo no presente estudo foi investigar a ocorrência de E. bieneusi em uma população de gatos domésticos em Campo Grande, Mato Grosso do Sul, Brasil. Sessenta amostras fecais de gatos diarréicos foram submetidas a reação em cadeia da polimerase (PCR) e os produtos de amplificação foram sequenciados para identificação molecular. E. bieneusi foi detectado em duas amostras (3,3%), ambos identificados como genótipo D. Esse genótipo tem sido relatado em animais e humanos e é considerado um genótipo zoonótico. Nossos resultados representam a primeira descrição de E. bieneusi em gatos domésticos no Brasil, reforçando a importância desse agente como fonte de infecção para animais e humanos.
Asunto(s)
Animales , Gatos , Enfermedades de los Gatos/diagnóstico , Microsporidiosis/veterinaria , Enterocytozoon/genética , Heces/microbiología , Brasil , Enfermedades de los Gatos/microbiología , Reacción en Cadena de la Polimerasa , Microsporidiosis/diagnóstico , Análisis de Secuencia de ADN , Enterocytozoon/aislamiento & purificación , GenotipoRESUMEN
Enterocytozoon bieneusi is an opportunistic intestinal pathogen that infects humans and a wide variety of animals worldwide. Our aim in this study was to investigate the occurrence of E. bieneusi in a domestic cat population in Campo Grande, Mato Grosso do Sul, Brazil. Sixty fecal samples from diarrheic cats were subjected to polymerase chain reaction (PCR) and the amplicons were sequenced for identification. E. bieneusi was detected in two samples (3.3%), both identified as genotype D. This genotype has already been reported in animals and humans and is considered a zoonotic genotype. Our findings represent the first report of E. bieneusi in domestic cats in Brazil, reinforcing the importance of identifying this agent as a source of infection in animals and humans.
Asunto(s)
Enfermedades de los Gatos/diagnóstico , Enterocytozoon/genética , Heces/microbiología , Microsporidiosis/veterinaria , Animales , Brasil , Enfermedades de los Gatos/microbiología , Gatos , Enterocytozoon/aislamiento & purificación , Genotipo , Microsporidiosis/diagnóstico , Reacción en Cadena de la Polimerasa , Análisis de Secuencia de ADNRESUMEN
BACKGROUND: Enterocytozoon bieneusi are the most common microsporidia associated with different clinical manifestations such as diarrhoea, respiratory tract inflammation and acalculous cholecystitis, especially in immunocompromised patients. Infection usually occurs by ingestion of food and water contaminated with spores, but can also result from direct contact with spores through broken skin, eye lesions, and sexual transmission, depending on the microsporidian species. Although there are reports of E. bieneusi found in humans and animals in Brazil, there are no published studies of environmental samples examined by molecular methods. OBJECTIVES: The purpose of this study was to verify the presence of E. bieneusi in raw sewage and treated effluent from a combined system by molecular methods. METHODS: Raw sewage and treated effluent samples collected from a combined system were analysed for the presence of E. bieneusi using the internal transcriber spacer (ITS) region of E. bieneusi by nested polymerase chain reaction. FINDINGS: The analysis revealed E. bieneusi presence and a novel genotype (EbRB) in one raw sewage sample and one treated effluent. MAIN CONCLUSIONS: The presence of E. bieneusi in final effluent indicates that the combined system may not remove microsporidian spores. This study is the first report of E. bieneusi in environmental samples in Brazil.
Asunto(s)
Enterocytozoon/genética , Enterocytozoon/aislamiento & purificación , Aguas del Alcantarillado/microbiología , Brasil , ADN de Hongos/genética , Genotipo , Filogenia , Reacción en Cadena de la Polimerasa , Análisis de Secuencia de ADNRESUMEN
BACKGROUND Enterocytozoon bieneusi are the most common microsporidia associated with different clinical manifestations such as diarrhoea, respiratory tract inflammation and acalculous cholecystitis, especially in immunocompromised patients. Infection usually occurs by ingestion of food and water contaminated with spores, but can also result from direct contact with spores through broken skin, eye lesions, and sexual transmission, depending on the microsporidian species. Although there are reports of E. bieneusi found in humans and animals in Brazil, there are no published studies of environmental samples examined by molecular methods. OBJECTIVES The purpose of this study was to verify the presence of E. bieneusi in raw sewage and treated effluent from a combined system by molecular methods. METHODS Raw sewage and treated effluent samples collected from a combined system were analysed for the presence of E. bieneusi using the internal transcriber spacer (ITS) region of E. bieneusi by nested polymerase chain reaction. FINDINGS The analysis revealed E. bieneusi presence and a novel genotype (EbRB) in one raw sewage sample and one treated effluent. MAIN CONCLUSIONS The presence of E. bieneusi in final effluent indicates that the combined system may not remove microsporidian spores. This study is the first report of E. bieneusi in environmental samples in Brazil.
Asunto(s)
ADN de Hongos/genética , Reacción en Cadena de la Polimerasa , Enterocytozoon/aislamiento & purificación , Enterocytozoon/genética , Filogenia , Brasil , Análisis de Secuencia , GenotipoRESUMEN
A total of 85 fecal samples from captive birds collected from October 2013 to September 2014 in Uberlândia and Belo Horizonte in the state of Minas Gerais (Brazil) were evaluated for the presence of Enterocytozoon bieneusi, Cryptosporidium, and Giardia by PCR. Of these, three birds were found positive for E. bieneusi (3.5%), two for Cryptosporidium (2.3%), and one for Giardia (1.2%). Two genotypes of E. bieneusi were detected by nucleotide sequence analysis of the ITS region, genotypes D and Peru 6 in a swan goose and in two rock pigeons, respectively. For Cryptosporidium and Giardia, nucleotide sequence analysis of the SSU rRNA identified Cryptosporidium baileyi and Duck genotype in a swan goose and a mandarin duck, respectively, and Giardia duodenalis assemblage A in a toco toucon. Our results demonstrate that human-pathogenic E. bieneusi genotypes D and Peru6 and G. duodenalis assemblage A are present in captive birds in Brazil, corroborating their potential role as a source of human infection and environmental contamination.
Asunto(s)
Enfermedades de las Aves/microbiología , Enfermedades de las Aves/parasitología , Criptosporidiosis/parasitología , Cryptosporidium/aislamiento & purificación , Enterocytozoon/aislamiento & purificación , Giardia/aislamiento & purificación , Giardiasis/veterinaria , Microsporidiosis/veterinaria , Animales , Enfermedades de las Aves/epidemiología , Aves/microbiología , Aves/parasitología , Brasil/epidemiología , Cryptosporidium/clasificación , Cryptosporidium/genética , Enterocytozoon/clasificación , Enterocytozoon/genética , Heces/microbiología , Heces/parasitología , Genotipo , Giardia/clasificación , Giardia/genética , Giardiasis/parasitología , Humanos , Microsporidiosis/microbiología , Reacción en Cadena de la PolimerasaRESUMEN
The presence of Enterocytozoon bieneusi in sheep has been reported in only three countries worldwide. The present study has found E. bieneusi in Brazilian sheep for the first time; in 24/125 (19.2%) fecal samples by PCR and on 8/10 (80%) farms from three diverse locations. A significantly greater number of lambs (34.1%) were found infected than older sheep (11.1%) (P=0.0036); most of the lambs were less than 6months of age. Farms with an intensive production system had a lower infection rate (10.5%) of infection than semi-intensive farms (23%), but this difference was not statistically significant. Sequencing analysis of the internal transcribed spacer (ITS) region of the rRNA gene revealed four known E. bieneusi genotypes (BEB6, BEB7, I, and LW1) and two novel genotypes (BEB18 and BEB19). Genotypes LW1 and BEB19 clustered within designated zoonotic Group 1 while genotypes BEB6, BEB7, I, and BEB18, and clustered within Group 2. BEB6 was the most prevalent (45.8%), followed by BEB7 (33.3%). Genotypes BEB6, I, and LW1 are zoonotic and can pose a risk to human health for immunocompromised individuals.
Asunto(s)
Enterocytozoon/genética , Microsporidiosis/veterinaria , Enfermedades de las Ovejas/microbiología , Envejecimiento , Animales , Brasil/epidemiología , Enterocytozoon/clasificación , Heces/parasitología , Genotipo , Humanos , Microsporidiosis/microbiología , Filogenia , Reacción en Cadena de la Polimerasa , Prevalencia , Ovinos , Enfermedades de las Ovejas/epidemiología , ZoonosisRESUMEN
A total of 151 fecal specimens from chickens were randomly collected from local markets in Uberlândia and Belo Horizonte in the state of Minas Gerais, Brazil, to evaluate the presence of Enterocytozoon bieneusi by polymerase chain reaction (PCR). Enterocytozoon bieneusi was identified in 24 fecal samples (15.9%). This represents the first report of E. bieneusi in chickens in Brazil. All PCR-positive specimens were sequenced and 4 genotypes were identified, Peru 6, Peru 11, Type IV, and D. All four genotypes have previously been reported as human pathogens and are potentially zoonotic. Our results demonstrate that human-pathogenic E. bieneusi genotypes are present in chickens in Brazil, corroborating their potential role as a source of human infection and environmental contamination.
Asunto(s)
Enterocytozoon/genética , Microsporidiosis/veterinaria , Enfermedades de las Aves de Corral/parasitología , Animales , Brasil/epidemiología , Pollos , ADN Espaciador Ribosómico/genética , Enterocytozoon/aislamiento & purificación , Variación Genética , Genotipo , Humanos , Microsporidiosis/diagnóstico , Microsporidiosis/epidemiología , Microsporidiosis/parasitología , Enfermedades de las Aves de Corral/diagnóstico , Enfermedades de las Aves de Corral/epidemiología , PrevalenciaRESUMEN
Microsporidia are widely recognized as important human pathogens with Enterocytozoon bieneusi as the most common species infecting humans and animals, including cattle. Although Brazil has the second largest cattle herd in the world and it is the largest exporter of beef there are no data on the presence or impact of E. bieneusi on this important population. To fill this knowledge gap, fecal specimens were collected from 452 cattle from pre-weaned calves to adult cattle in the state of Rio de Janeiro. Host factors including age, gender, dairy/beef, body composition, and fecal consistency were included in the study. Using molecular methods, E. bieneusi was found in 79/452 (17.5%) fecal specimens. This represents the first report of this parasite in Brazilian cattle. A significantly higher prevalence was found in calves less than 2 months of age (27.6%) and those 3-8 months of age (28.8%) versus heifers (14.1%) and adults (1.4%) (P<0.05). Dairy cattle (26.2%) had a higher prevalence than beef cattle (9.7%) (P<0.001). No correlation was found between infection and gender, body composition, and fecal consistency. Molecular characterization of the internal transcribed spacer (ITS) revealed 12 genotypes; five previously reported in cattle (BEB4, BEB8, D, EbpA and I), and seven novel genotypes (BEB11-BEB17). A phylogenetic analysis showed that 6 genotypes (D, EbpA, BEB12, BEB13, BEB15, and BEB16) identified in 18 animals clustered within the designated zoonotic Group 1 while the other 6 genotypes (I, BEB4, BEB8, BEB11, BEB14, BEB17) identified in 61 animals clustered within Group 2. The identification of genotypes in Brazilian cattle that have previously been reported in humans highlights the potential risk of zoonotic transmission and suggests that the role of cattle in transmission of human infections requires further study.
Asunto(s)
Enfermedades de los Bovinos/parasitología , Enterocytozoon/aislamiento & purificación , Microsporidiosis/veterinaria , Distribución por Edad , Animales , Secuencia de Bases , Brasil/epidemiología , Bovinos , Enfermedades de los Bovinos/epidemiología , Enterocytozoon/clasificación , Enterocytozoon/genética , Heces/química , Heces/parasitología , Femenino , Genotipo , Masculino , Microsporidiosis/epidemiología , Microsporidiosis/parasitología , Filogenia , PrevalenciaRESUMEN
Although Brazil is the world's fourth largest producer and exporter of pork, there is no information on Enterocytozoon bieneusi in pigs. This study was undertaken to determine the presence of E. bieneusi in pigs in the State of Rio de Janeiro, Brazil. Fecal samples were collected from 91 pigs (1- to 12-month-old) in 10 properties and examined by molecular methods. The presence of E. bieneusi was determined by PCR and all PCR positive specimens were sequenced to determine the genotype by nucleotide sequence analysis of the internal transcribed spacer of the rRNA gene. E. bieneusi was found in pigs in all farms. Fifty four (59.3%) samples were E. bieneusi-positive. A wide genetic diversity was found with 21 genotypes identified, 4 previously reported (O, EbpA, CS-1, and H) and 17 novel genotypes named PigEb1-PigEb17. All 17 novel genotypes identified in this study clustered within the previously designated zoonotic Group 1. The most prevalent genotypes were novel genotypes PigEb2 and PigEb4 (16/91, 17.6%, each). Mixed infections with 2 or 3 genotypes were detected in 13 pigs (24.1%). The high prevalence in pigs observed in this study, the description of two known zoonotic genotypes (EbpA and O), and the report of 17 new genotypes of E. bieneusi, represent an important advancement in the study of the wide genetic diversity of this organism, emphasizing the importance of further research, especially in geographical areas where little or no research has been conducted. The zoonotic risk of these novel genotypes and their importance to other animal species is still unknown, but needs to be further evaluated.
Asunto(s)
Enterocytozoon/aislamiento & purificación , Heces/microbiología , Ganado/microbiología , Microsporidiosis/veterinaria , Sus scrofa/microbiología , Enfermedades de los Porcinos/microbiología , Animales , Secuencia de Bases , Brasil/epidemiología , Coinfección/microbiología , Coinfección/veterinaria , ADN de Hongos/genética , ADN Espaciador Ribosómico/genética , Enterocytozoon/genética , Variación Genética , Genotipo , Humanos , Microsporidiosis/epidemiología , Microsporidiosis/microbiología , Datos de Secuencia Molecular , Filogenia , Reacción en Cadena de la Polimerasa , Prevalencia , Análisis de Secuencia de ADN , Porcinos , Enfermedades de los Porcinos/epidemiologíaRESUMEN
Microsporidia are ubiquitous parasites infecting all animal phyla and we present evidence that supports their zoonotic potential. Fecal samples taken from domestic (cats and dogs), farm (pigs, rabbits and ostriches) and wild animals (foxes) from different provinces of Spain were evaluated for microsporidia infection by light microscopy and PCR. After Microsporidia species identification, E. bieneusi genotypes were additionally studied by sequence analysis of the ITS region. Eighty-five samples out of 159 exhibited structures that were compatible with microsporidia spores by Webers stain with 37 of them being confirmed by PCR. Microsporidia species identified included E. bieneusi, E. intestinalis and A. algerae. We report the first diagnosis of E. intestinalis and E. bieneusi in ostriches and A. algerae in pigs. We also provide new information on the molecular characterization of E. bieneusi isolates both in rabbits and ostriches. All of the E. bieneusi genotypes identified belonged to the zoonotic group of genotypes (Group I) including genotypes A (dogs), I (pigs), D (rabbits and foxes) and type IV (ostriches). Our results demonstrate that microsporidia are present in domestic, farm and wild animals in Spain, corroborating their potential role as a source of human infection and environmental contamination.
Asunto(s)
Enterocytozoon/genética , Enterocytozoon/aislamiento & purificación , Técnicas de Genotipaje , Genotipo , Geografía , Humanos , Microsporidiosis/veterinaria , Datos de Secuencia Molecular , España , Especificidad de la EspecieRESUMEN
Fecal specimens were obtained from a total of 70 dairy calves less than two months old on 11 municipalities in Buenos Aires, Argentina. After removal of fecal debris by sieving and sucrose flotation, specimens were subjected to PCR to detect the presence of Enterocytozoon bieneusi. PCR revealed a 14.3% of prevalence for E. bieneusi with 10 positive calves from 7 municipalities. Gene sequence analysis conducted in all samples positives by PCR revealed the presence of six genotypes; four previously reported in cattle as well as humans (D, I, J, and BEB4), one never reported in cattle before but previously reported in humans (EbpC), and one novel genotype (BEB10). These results constitute the first molecular characterization of E. bieneusi in Argentina, and suggest a potential risk of zoonotic transmission in this area.
Asunto(s)
Enfermedades de los Bovinos/microbiología , Enterocytozoon/fisiología , Microsporidiosis/veterinaria , Animales , Argentina/epidemiología , Bovinos , Enfermedades de los Bovinos/epidemiología , ADN Espaciador Ribosómico/genética , Industria Lechera , Enterocytozoon/genética , Enterocytozoon/aislamiento & purificación , Heces/microbiología , Genotipo , Humanos , Microsporidiosis/epidemiología , Microsporidiosis/microbiología , Datos de Secuencia MolecularRESUMEN
The population structure of Enterocytozoon bieneusi was examined by multilocus sequence typing (MLST) of 64 specimens from AIDS patients in Peru, Nigeria, and India and five specimens from captive baboons in Kenya using a combination of the ribosomal internal transcribed spacer (ITS) and four microsatellite and minisatellite markers. Parasites in different geographic locations (Peru, India, and Nigeria) all had strong and significant linkage disequilibrium (LD) and only limited recombination, indicative of a clonal population structure in E. bieneusi from each location. When isolates of various geographical areas were treated as a single population, phylogenetic analysis and substructural analysis using STRUCTURE found no evidence for the existence of geographically segregated sub-populations. Nevertheless, both analyses revealed the presence of two major genetically isolated groups of E. bieneusi: one (sub-population 1) contained all isolates of the anthroponotic ITS genotype A, whereas the other (sub-population 2) harbored isolates of multiple ITS genotypes with zoonotic potential. This was also supported by FST analysis. The measurement of LD and recombination rates indicated that sub-population 2 had a clonal population structure, whereas sub-population 1 had an epidemic population structure. The data confirmed the existence of genetic sub-populations in E. bieneusi that may be transmitted differently in humans.
Asunto(s)
Enterocytozoon/clasificación , Enterocytozoon/genética , Tipificación de Secuencias Multilocus , Adulto , Animales , Niño , ADN Intergénico/genética , Frecuencia de los Genes , Genética de Población , Genotipo , Humanos , India , Kenia , Datos de Secuencia Molecular , Nigeria , Perú , Filogenia , Filogeografía , Polimorfismo GenéticoRESUMEN
Enterocytozoon bieneusi was detected in dogs in Bogota, Colombia for the first time. Of 120 dogs, 15% were positive by polymerase chain reaction (PCR). Infected dogs ranged from 2 to 14 years of age, and more male than female dogs were infected with E. bieneusi (20% versus 5%). All PCR-positive specimens were sequenced and three genotypes were identified. A dog-specific genotype (PtEbIX) was identified in feces from 16 dogs. For two dogs, the nucleotide sequences obtained were genetically identical to the E. bieneusi Peru 5 and K genotypes previously reported as human pathogens. This is the first time that Peru 5 and K genotypes have been identified in dogs. This study confirms that dogs are infected with both human pathogenic and host-specific genotypes.
Asunto(s)
Enfermedades de los Perros/microbiología , Enterocytozoon/genética , Microsporidiosis/veterinaria , Animales , Colombia/epidemiología , ADN de Hongos/química , Enfermedades de los Perros/epidemiología , Perros , Enterocytozoon/clasificación , Enterocytozoon/aislamiento & purificación , Heces/microbiología , Femenino , Genotipo , Masculino , Microsporidiosis/epidemiología , Microsporidiosis/microbiología , Reacción en Cadena de la Polimerasa/veterinaria , Prevalencia , Alineación de Secuencia/veterinaria , Análisis de Secuencia de ADN/veterinariaRESUMEN
This study involved ninety five formalin-fixed paraffin-embedded duodenal biopsy specimens retrieved from hospital files that were microscopically observed for the presence of microsporidia. Eleven samples that revealed compatible organisms were analyzed by the polymerase chain reaction (PCR) with four different protocols for the detection of Enterocytozoon bieneusi. Amplicons of the right size were obtained by at least one method for nine samples, remaining two negative ones. We report a PCR methodology that allows the use of archival specimens obtained for traditional pathology.