ABSTRACT
We aimed to assess the risks of
Subject(s)
Humans , China , Cryptosporidiosis/microbiology , Cryptosporidium/isolation & purification , Giardia/isolation & purification , Giardiasis/microbiology , Risk Assessment , Water Microbiology , Water Supply/statistics & numerical dataABSTRACT
ABSTRACT The study was conducted to compare the specificity of immunological diagnostic methods used for the diagnosis of Cryptosporidium species capable of causing life-threatening infection in both immunosuppressed and immunocompetent patients. For the detection of Cryptosporidium species in 79 animals with diarrhoea, we used three Copro-antigen tests: RIDASCREEN® Cryptosporidium test, Cryptosporidium 2nd Generation (ELISA) and RIDA®QUICK Cryptosporidium. For immunoassays we used positive and negative samples detected by means of polymerase chain reaction and validated by sequencing and nested polymerase chain reaction to confirm the presence six different species of Cryptosporidium species. Prevalence of cryptosporidiosis in the entire group determined by enzyme immunoassay, enzyme linked immunosorbent assay, immuno-chromatographic test and polymerase chain reaction was 34.17%, 27.84%, 6.33% and 27.84%, respectively. Sensitivity of animal samples with enzyme immunoassay, enzyme linked immunosorbent assay, and immuno-chromatographic test was 63.6%, 40.9% and 22.7%, resp., when questionable samples were considered positive, whereas specificity of enzyme immunoassay, enzyme linked immunosorbent assay and immuno-chromatographic test was 75.9%, 78.9% and 100%, respectively. Positive predictive values and negative predictive values were different for all the tests. These differences results are controversial and therefore reliability and reproducibility of immunoassays as the only diagnostic method is questionable. The use of various Cryptosporidium species in diagnosis based on immunological testing and different results obtained by individual tests indicate potential differences in Copro-antigens produced by individual Cryptosporidium species.
Subject(s)
Animals , Immunologic Tests/methods , Cryptosporidiosis/microbiology , Cryptosporidium/isolation & purification , Diarrhea/veterinary , Immunologic Tests/economics , Immunologic Tests/veterinary , Sensitivity and Specificity , Cryptosporidiosis/diagnosis , Cryptosporidium/genetics , Cryptosporidium/immunology , Diarrhea/diagnosis , Diarrhea/microbiologyABSTRACT
The identification and characterisation of Cryptosporidiumgenotypes and subtypes are fundamental to the study of cryptosporidiosis epidemiology, aiding in prevention and control strategies. The objective was to determine the genetic diversity ofCryptosporidium in samples obtained from hospitals of Rio de Janeiro, Brazil, and Buenos Aires, Argentina. Samples were analysed by microscopy and TaqMan polymerase chain reaction (PCR) assays forCryptosporidium detection, genotyped by nested-PCR-restriction fragment length polymorphism (RFLP) analysis of the 18S rRNA gene and subtyped by DNA sequencing of the gp60 gene. Among the 89 samples from Rio de Janeiro, Cryptosporidium spp were detected in 26 by microscopy/TaqMan PCR. In samples from Buenos Aires,Cryptosporidium was diagnosed in 15 patients of the 132 studied. The TaqMan PCR and the nested-PCR-RFLP detected Cryptosporidium parvum, Cryptosporidium hominis, and co-infections of both species. In Brazilian samples, the subtypes IbA10G2 and IIcA5G3 were observed. The subtypes found in Argentinean samples were IbA10G2, IaA10G1R4, IaA11G1R4, and IeA11G3T3, and mixed subtypes of Ia and IIa families were detected in the co-infections. C. hominis was the species more frequently detected, and subtype family Ib was reported in both countries. Subtype diversity was higher in Buenos Aires than in Rio de Janeiro and two new subtypes were described for the first time.
Subject(s)
Adolescent , Adult , Aged , Child , Child, Preschool , Female , Humans , Infant , Male , Middle Aged , Young Adult , Cryptosporidiosis/microbiology , Cryptosporidium/genetics , DNA, Protozoan/genetics , Genetic Variation/genetics , Argentina , Brazil , Cryptosporidium/classification , DNA, Ribosomal/genetics , Genotype , Polymorphism, Restriction Fragment Length , Sequence Analysis, DNAABSTRACT
Informar la experiencia de erradicación de Cryptosporidium y Microsporidium spp., en pacientes inmunocomprometidos con diarrea crónica utilizando nitazoxanida como terapia. Se evaluaron 50 pacientes con diarrea crónica (38 mujeres y 12 hombres), edad promedio de 45 y 56 años respectivamente; mayores de edad, dieron su consentimiento informado por escrito; con diagnósticos de cáncer de vías digestivas (veintidos), mama (diez y seis), cuello uterino (siete) y próstata (cinco); además 6 presentaban infección por VIH. Se hizo diagnóstico parasitológico empleando la coloración de Kinyoun. Recibieron nitazoxanida 500 mg/VO/BID por períodos de 3,6,9 días dependiendo del resultado de los controles microbiologicos. Se identificó como agente único a Cryptosporidium spp. en 2 pacientes y Microsporidium sp., en 6 pacientes, y combinados en 42 pacientes. Al tercer día de tratamiento, en 30 pacientes se logró erradicación de la criptosporidiosis, persistiendo con clínica diarreica en 28 de ellos; al sexto día se evidenció cura microbiológica (erradicación parasitaria de ambos microorganismos) y clínica (cese diarrea), en un total de 40 pacientes; al noveno día de tratamiento, se logró cura clínica y microbiológica en todos los pacientes incluyéndose en este último grupo todos los pacientes con infección VIH. Se evidenció epigastralgia en 4 pacientes que recibieron 9 días de terapia. Nitazoxanida parece ser un tratamieto eficaz y seguro en diarrea crónica micro y/o criptosporidiana en pacientes inmunocomprometidos oncológicos y es necesario realizar pesquisa microbiológica de parasitosis emergente y prolongación del tratamiento más allá de losa 3 días convencionalmente recomendados.
Subject(s)
Humans , Male , Adult , Female , Middle Aged , Cryptosporidiosis/microbiology , Diarrhea/diagnosis , Diarrhea/therapy , Microsporidia/immunology , Digestive System Neoplasms/etiology , Digestive System Neoplasms/pathology , Parasitology/methods , Infectious Disease Medicine , Medical OncologyABSTRACT
Amostras fecais de indivíduos portadores de HIV, e com Aids, foram submetidas ao diagnóstico laboratorial de criptosporidiose. Do total de 29 amostras, 15 apresentaram oocistos de Cryptosporidium sp no exame parasitológico, 13 foram reativas em ELISA para detecção de coproantígenos e em 16 foram obtidos produtos de amplificação por Nested-PCR. Foi realizada caracterização genotípica em cinco destes produtos amplificados, por meio de PCR-RFLP com a enzima de restrição AfaI (RsaI). Dois isolados foram identificados como Cryptosporidium hominis, um como Cryptosporidium parvum e dois apresentaram perfil eletroforético correspondente a Cryptosporidium meleagridis. Esses achados trazem evidências de que diferentes espécies de Cryptosporidium circulam no ambiente, traduzindo-se em risco tanto para humanos quanto para outros animais.
Subject(s)
Cryptosporidiosis/microbiology , Cryptosporidium/isolation & purification , Acquired Immunodeficiency Syndrome/parasitology , GenotypeABSTRACT
Cryptosporidium parvum oocysts were isolated from a child suffering from acute gastroenteritis and successfully passaged in a calf and mice (designated hereafter SNU-H1) in the Republic of Korea; its molecular genotype has been analyzed. The GAG microsatellite region was amplified by a polymerase chain reaction (PCR), with a 238 base pair product, which is commonly displayed in C. parvum. The isolate was shown to be a mixture of the genotypes 1 (anthroponotic) and 2 (zoonotic). To study its infectivity in animals, 2 calves and 3 strains of mice were infected with the SNU-H1; in these animals, the propagation of both genotypes was successful. In immunosuppressed (ImSP) BALB/c and C57BL/6 mice the number of oocysts decreased after day 10 post-infection (PI) ; but in ImSP ICR mice, they remained constant until day 27 PI. The results show that both the C. parvum genotypes 1 and 2 can be propagated in calves and ImSP mice.
Subject(s)
Animals , Cattle , Child , Humans , Male , Mice , Cryptosporidiosis/microbiology , Cryptosporidium parvum/genetics , Diarrhea/parasitology , Feces/parasitology , Genotype , Korea , Mice, Inbred BALB C , Mice, Inbred C57BL , Mice, Inbred ICR , Oocysts , Polymerase Chain Reaction , Zoonoses/parasitologyABSTRACT
Se examinaron durante un año 300 muestras de materia fecal de 210 niños diarreicos, para la búsqueda de Cryptosporidium sp., procedentes del Hospital Provincial del Centenario, Rosario. El rango de edad de los niños osciló entre la primer semana de vida y los 13 años, aunque su mayoría no superó los 3 años. Las heces se recolectaron en formol 10%, se enriquecieron por el método de formol-eter y se colorearon con las técnicas permanentes se Safranina 1% y Ziehl Neelsen modificada. Los ooquistes de Cryptosporidium sp. fueron detectados en 16 de los 210 niños examinados (7,6%). La cantidad de ooquistes presentes en las muestras positivas fue de moderada a abundante, con excepción de un solo niño donde el número de ooquistes fue demasiado escaso. Se obtuvieron muestras adicionales de sólo 5 de estos 16 niños. En 3 de ellos la segunda muestra se negativizó. En ninguna de las muestras positivas se detectaron simultáneamente trofozoitos, quistes, esporoquistes, huevos o larvas de otro enteoparásito. La búsqueda de Cryptosporidium sp. En niños diarreicos debe ser considerada como rutina parasitológica para el diagnóstico etiológico diferencial
Subject(s)
Humans , Child , Child, Preschool , Infant , Infant, Newborn , Adolescent , Animals , Coccidia/isolation & purification , Cryptosporidiosis/microbiology , Cryptosporidium/isolation & purification , Feces/microbiologyABSTRACT
Se señala la presencia de Cryptosporidium sp. en 14 casos humanos por primera vez en el Perú, sobre un total de 2.200 muestras de heces examinadas y se describe la forma de hallazgo, utilizando coloración y concentración