Caracterización molecular de aislados humanos de Cryptosporidium spp. procedentes de 2 diferentes localizaciones de España / Molecular characterization of Cryptosporidium spp. isolated in humans in two different locations in Spain

Las técnicas de diagnóstico molecular por PCR permiten distinguir entre las diferentes especies de Cryptosporidium morfológicamente idénticas capaces de infectar a humanos. De las 23 especies actualmente reconocidas en el género, al menos 9 son capaces de infectar a humanos. Por ello, y debido a que la intensidad de las manifestaciones clínicas, la patogenicidad, la excreción de ooquistes y la incidencia varían entre ellas, la realización de estudios moleculares es crucial para una mejor comprensión de la epidemiología de la criptosporidiosis humana. En el presente trabajo se analizan muestras procedentes de 2 estudios independientes: uno formado por 23 muestras procedentes de Madrid y otro compuesto por 72 muestras procedentes de La Coruña, todas ellas positivas para Cryptosporidium spp. por métodos microscópicos y pertenecientes a casos aislados de criptosporidiosis. Para la identificación a nivel de especie se utilizaron las regiones de diagnóstico descritas para el ADNr 18S y las regiones de diagnóstico del gen de la COWP. De las 95 muestras analizadas, se consiguió extraer y amplificar ADN en 77 casos, en los que las especies causantes de la infección fueron: C. parvum (40 casos: 2 Madrid y 38 La Coruña), C. hominis (30 casos: 10 Madrid y 20 La Coruña) y C. meleagridis (2 casos: uno Madrid y uno La Coruña). En otros 5 casos fue imposible detectar la especie responsable de la infección, aunque se confirmara su positividad por PCR (4 Madrid y uno La Coruña). Los genotipos aislados en estos pacientes se correlacionaron con los hallados en animales de las mismas regiones (AU)

Molecular PCR based diagnostic techniques have enabled us to distinguish between the different, morphologically identical, Cryptosporidium species that can infect humans. Of the 23 recognized species in the genus, at least 9 are able to infect humans. As the intensity of the clinical manifestations, pathogenicity, excretion of oocysts, and incidence, are different between this species, molecular studies are crucial for a better understanding of the epidemiology of human cryptosporidiosis. Samples form two independent studies are analyzed in this publication. One included 23 samples from Madrid, and the other, 72 samples from La Coruña. All of them positive for Cryptosporidium spp. by microscopic methods and belonging to isolated cases of human cryptosporidiosis. For the identification of the species responsible for the infection, the 18S rDNA diagnostic region and the COWP gene diagnostic regions were used. Out of the 95 samples tested, in 77 cases we were able to extract and amplify DNA. In those cases the species responsible for the infection were: C. parvum (40 cases, 2 Madrid and 38 La Coruña), C. hominis (30 cases, 10 Madrid and 20 La Coruña) and C. meleagridis (2 cases, 1 Madrid and 1 La Coruña). In 5 samples it was impossible to detect the species responsible for the infection, but their positivity was confirmed by PCR (4 Madrid and 1 La Coruña). The genotypes of the isolates from patients correlated well with animals from the same regions

[Molecular characterization of Cryptosporidium spp. isolated in humans in two different locations in Spain]. / Caracterización molecular de aislados humanos de Cryptosporidium spp. procedentes de 2 diferentes localizaciones de España.

Molecular PCR based diagnostic techniques have enabled us to distinguish between the different, morphologically identical, Cryptosporidium species that can infect humans. Of the 23 recognized species in the genus, at least 9 are able to infect humans. As the intensity of the clinical manifestations, pathogenicity, excretion of oocysts, and incidence, are different between this species, molecular studies are crucial for a better understanding of the epidemiology of human cryptosporidiosis. Samples form two independent studies are analyzed in this publication. One included 23 samples from Madrid, and the other, 72 samples from La Coruña. All of them positive for Cryptosporidium spp. by microscopic methods and belonging to isolated cases of human cryptosporidiosis. For the identification of the species responsible for the infection, the 18S rDNA diagnostic region and the COWP gene diagnostic regions were used. Out of the 95 samples tested, in 77 cases we were able to extract and amplify DNA. In those cases the species responsible for the infection were: C. parvum (40 cases, 2 Madrid and 38 La Coruña), C. hominis (30 cases, 10 Madrid and 20 La Coruña) and C. meleagridis (2 cases, 1 Madrid and 1 La Coruña). In 5 samples it was impossible to detect the species responsible for the infection, but their positivity was confirmed by PCR (4 Madrid and 1 La Coruña). The genotypes of the isolates from patients correlated well with animals from the same regions.

[Cryptosporidium: a genus in revision. The situation in Spain]. / Cryptosporidium: un género en revisión. Situación en España.

Genus Cryptosporidium, has undergone major revisions in recent years. The identification of new species and their major reservoirs has contributed to the knowledge of the epidemiology of human infection. In Spain, although there are many publications, few studies have been conducted to identify the circulating species and genotypes. This fact has led us to review and update these new studies published in Spain, particularly those that use molecular methods in order to characterise the species and genotypes present in our country.

Cryptosporidium: un género en revisión. Situación en España / Cryptosporidium: a genus in revision. The Situation in Spain

El género Cryptosporidium, ha sido objeto de importantes revisiones en los últimos años, tanto a nivel taxonómico, con la identificación de nuevas especies y sus principales reservorios, como por la contribución de esta información al conocimiento de la epidemiología de la infección en humanos. En España, aunque son múltiples las publicaciones realizadas, todavía son pocos los estudios llevados a cabo para la identificación de las especies y genotipos circulantes. Este hecho nos ha motivado a realizar una revisión actualizando estas novedades y los estudios publicados en España, especialmente aquellos donde se utilicen métodos moleculares que permiten la caracterización de especies y genotipos presentes en nuestro país (AU)

Genus Cryptosporidium, has undergone major revisions in recent years. The identification of new species and their major reservoirs has contributed to the knowledge of the epidemiology of human infection. In Spain, although there are many publications, few studies have been conducted to identify the circulating species and genotypes. This fact has led us to review and update these new studies published in Spain, particularly those that use molecular methods in order to characterise the species and genotypes present in our country (AU)

Detection and molecular characterization of Cryptosporidium bovis-like isolate from a newborn lamb in Spain.

Species of Cryptosporidium infect a broad variety of animals. Because morphological features of the secreted oocysts are not useful in identifying the parasite at the species level, molecular tools were used to accomplish this task, leading to discovery of new Cryptosporidium species. With the use of this approach, Cryptosporidium bovis has recently been described as a new species infecting bovines and several other hosts, but clearly distinct from C. parvum. In this report, we present a description of a Cryptosporidium sp. isolate from a newborn lamb from a farm in Spain. The isolate seemed to be very similar to C. bovis based on the analysis of the gene that codes for the 18S rRNA.

Frecuencia de microsporidiosis intestinal en pacientes positivos para VIH mediante las técnicas de Gram cromotropo rápido y PCR / Frequency of intestinal microsporidian infections in HIV-positive patients, as diagnosis by quick hot Gram chromotrope staining and PCR

Los microsporidios son protozoos intracelulares obligados, implicados en procesos de diarrea persistente en pacientes con sida, aunque no son exclusivos de este grupo de pacientes. La prevalencia de microsporidios en diferentes países varía entre 8 por ciento y 52 por ciento. En nuestro medio no se conoce su frecuencia, por lo que este trabajo se propuso determinar la frecuencia de microsporidiosis intestinal en pacientes positivos para VIH, mediante la prueba del Gram cromotropo rápido ( quick hot Gram) y la PCR; para esto se realizó un estudio prospectivo, descriptivo, con una población intencional de todos los pacientes positivos para VIH remitidos al Laboratorio del Grupo Interdisciplinario para el Estudio de las Parasitosis Intestinales por las diferentes instituciones de atención de pacientes positivos para VIH de Medellín en el periodo comprendido entre agosto de 2001 y septiembre de 2002. Se hizo una encuesta clínico-epidemiológica y se practicaron análisis coprológicos seriados que incluían examen directo, por concentración y tinciones especiales para coccidias y microsporidios intestinales; además, se solicitó recuento de linfocitos TCD4+ y carga viral. Se estudiaron 103 pacientes en edades comprendidas entre 2 y 74 años; el 70 por ciento(72/103) presentaba diarrea al ingreso al estudio; la mayoría (83,5 por ciento) fueron hombres. La frecuencia global de microsporidiosis intestinal fue de 3,9 por ciento(4/103); se encontraron tres pacientes positivos para Enterocytozoon bieneusi y uno con Encephalitozoon intestinalis; otras parasitosis intestinales representaron el 39,8 por ciento. La frecuencia de microsporidiosis en este estudio fue relativamente baja; además, como era de esperarse, la mayoría de los casos de microsporidios estuvieron asociados con diarrea prolongada y recuentos de LTCD4+ menores de 100 cél/µl y cargas virales superiores a 100.000 copias (3/4)

[Frequency of intestinal microsporidian infections in HIV-positive patients, as diagnosis by quick hot Gram chromotrope staining and PCR]. / Frecuencia de microsporidiosis intestinal en pacientes positivos para VIH mediante las técnicas de Gram cromotropo rápido y PCR.

Microsporidia are intracellular obligate parasites, today mainly associated with diarrhea in AIDS patients. Microsporidia prevalence ranges from 8% to 52% in different countries, as evaluated by several diagnostic methods, such as the stain test and PCR. In Medellín, Colombia, its frequency is unknown, and hence, a study was undertaken to determine the frequency of intestinal microsporidiosis in HIV patients, by means of the quick-hot Gram chromotrope test and the PCR. A prospective and descriptive study of an intentional population of all HIV-positive patients was sent to the Grupo Interdisciplinario para el Estudio de las Parasitosis Intestinales laboratory by institutions treating the HIV-positive patients of Medellín between August 2001 and September 2002. The clinical-epidemiological survey included a serial stool test with direct concentration and special stains for coccidiae and intestinal microsporidia. In addition, counts of lymphocytes TCD4+ and viral load were requested. One hundred and three patients with ages ranging from 2-74 years were evaluated. Seventy percent presented with diarrhea--mostly in men (83.5%). The overall frequency of intestinal microsporidiosis was 3.9% and that of other intestinal parasitic infections was 39.8%. Three of the four patients positive for microsporida were infected with Enterocytozoon bieneusi and one with Encephalitozoon intestinalis. The microsporidiosis frequency was relatively low with 3 of the 4 cases associated with protracted diarrhea, counts of LTCD4+ below 100 cel/microl and viral loads up to 100,000 copies.