Infestación por Cryptosporidium Spp e Isospora Belli en preescolar inmunocompetente: a propósito de un caso / Infestation by Crytosporidium Spp and Isospora belli in immunocompetent preschool: a report of a case

El Cryptosporidium spp e Isospora belli son parásitos emergentes, que representan la cuarta causa de diarrea a nivel mundial, principalmente en niños y en pacientes inmunocomprometidos. Producen diarrea aguda o crónica dependiendo de la edad del paciente, estado nutricional e inmunológico asociado a factores sanitarios desfavorables. El diagnostico se realiza por visualización directa en heces con tinción de Zelh Neelsen modificado o Kinyou y biopsia intestinal con presencia de protozoos en las criptas y atrofia vellositaria de acuerdo al grado de infestación. Se reporta el caso de preescolar de 2 años de edad, eutrófico e inmunocompetente, perteneciente a estrato social bajo; con episodios de diarreas acuosas autolimitadas, dolor y distensión abdominal frecuentes. La biopsia intestinal revelo atrofia vellositaria e infestación simultanea por Cryptosporidium spp e Isospora belli corroborado por Tinción de Kinyou en heces; se descarto además Alergia Alimentaria, Enfermedad Celiaca e Inmunodeficiencias. El propósito de este caso clínico es alertar sobre la necesidad de incluir dentro del protocolo de estudio de diarrea crónica, la búsqueda de protozoarios formadores de esporas, mediante tinción especial en heces; un método no invasivo y sencillo, no solicitado en forma rutinaria.

Cryptosporidium spp and Isospora belli parasites are emerging that represent the fourth leading cause of diarrhea worldwide, mainly in children and in immunocompromised patients. Acute or chronic diarrhea occur depending on the patient's age, nutritional status and immunological factors associated with adverse health. The diagnosis is made by direct visualization in feces Neelsen stain Zelh Kinyou modified or intestinal biopsy and the presence of protozoa in the crypts and villous atrophy according to the degree of infestation. We report the case of preschool age 2, eutrophic immunocompetent, belonging to low socioeconomic levels, with self-limiting episodes of acute watery diarrhea, frequent abdominal pain and bloating. The intestinal biopsy revealed villous atrophy and simultaneous infestation by Cryptosporidium spp and Isospora belli Kinyou confirmed by staining in feces, while discarding also Food Allergy, Celiac disease and immunodeficiencies. The purpose of this case to alert about the need to include in the study protocol of chronic diarrhea, the search for spore-forming protozoa by special staining in feces, a noninvasive and simple method, not routinely requested.

Anticorpos anti-intiminas α, β, e γ de Escherichia coli em soros e colostros de adultos saudáveis da Grande São Paulo / Antibodies reactive with α, β,and γ intimins of Escherichia coli in serum and colostrum samples of healthy adults living in São Paulo, Brazil

A diarréia é um importante problema de saúde pública no mundo inteiro e a Escherichía coli é um dos mais freqüentes microorganismos causadores desta doença. A Escherichia coli enteropatogênica (EPEC), um dos principais agentes etiológicos das diarréias infantis no nosso país, é genética e fenotipicamente relacionada com a E. colí enterohemorrágica (EHEC) que além de provocar diarréia é responsável por complicações como síndrome hemolítica urêmica (HUS) e colite hemorrágica (HC). Embora a EHEC seja considerada emergente pela OMS, no Brasil poucos casos de complicações como HUS e HC foram reportados. O mecanismo de patogenicidade comum entre EPEC e EHEC é conhecido como a lesão "attaching and effacing" nos microvilos do enterócito. Esta lesão é mediada por um conjunto de fatores de virulência, dentre eles a intimina. A intimina é uma proteína de membrana externa, responsável pelo íntimo contato da bactéria com o enterócito, possui uma região N-terminal que é altamente conservada e uma região C-terminal que é variável. De acordo com a região variável, existem vários subtipos de intimina, dentre eles as intiminas , α, β e γ...

Successful treatment of rotavirus-induced diarrhoea in suckling mice with egg yolk immunoglobulin.

The role of specific immunoglobulins at mucosal sites in imparting protection against disease, such as rotavirus-associated diarrhoea, is well-established. Oral immunoglobulin therapy with egg yolk-derived anti-rotavirus immunoglobulins has previously been shown to achieve moderate therapeutic effect in diarrhoea due to rotavirus in a clinical trial. Here, data on the therapeutic potential of the same immunoglobulin preparation in an infant mouse model of rotavirus-induced diarrhoea is presented. The use of an animal model has allowed therapy to be evaluated with higher doses of immunoglobulins and has suggested that an improved therapeutic effect can be achieved by increasing the dose in the clinical setting.

Development of a monoclonal antibody-based co-agglutination test to detect enterotoxigenic Escherichia coli isolated from diarrheic neonatal calves

Escherichia coli (E. coli) strains were collected from young diarrheic calves in farms and field. Strains that expressed the K99 (F5) antigen were identified by agglutination tests using reference antibodies to K99 antigen and electron microscopy. The K99 antigen from a selected field strain (SAR-14) was heat-extracted and fractionated on a Sepharose CL-4B column. Further purification was carried out by sodium deoxycholate treatment and/or ion-exchange chromatography. Monoclonal antibodies to purified K99 antigen were produced by the hybridoma technique, and a specific clone, NEK99-5.6.12, was selected for propagation in tissue culture. The antibodies, thus obtained, were affinity-purified, characterized and coated onto Giemsastained Cowan-I strain of Staphylococcus aureus (S. aureus). The antibody-coated S. aureus were used in a coagglutination test to detect K99+ E. coli isolated from feces of diarrheic calves. The specificity of the test was validated against reference monoclonal antibodies used in co-agglutination tests, as well as in ELISA. Specificity of the monoclonal antibodies was also tested against various Gram negative bacteria. The developed antibodies specifically detected purified K99 antigen in immunoblots, as well as K99+ E. coli in ELISA and co-agglutination tests. The co-agglutination test was specific and convenient for large-scale screening of K99+ E. coli isolates.

Study of cryptosporidiasis on immune competent and immune suppressed children using antigen prepared from calves after experimental infection by Indirect Fluorescent Antibody Technique [IFAT]

An epidemiological study was done on 2 groups of children infected with watery diarrhea in some hospitals of Baghdad [Group 1: Immunocompetent outpatients. Group 2: Immunosuppressed inpatients]. Modified Cold Ziehl Neelsen stain was used in diagnosis. The whole oocysts antigen prepared after conduction of experimental infection in 2 calves by using isolated oocysts from children stool, and this prepared antigen was used in Indirect Fluorescent Antibody Technique [IFAT]. The percentage of infection in the 2 groups of outpatients and inpatients were 21 2% and 32% respectively, and there was a difference in percentage of infection between the 3 groups of children, the highest was 36.4% in group infected with leukemia and received chemical and radiotherapy. There was no significant difference noticed between the two sexes. The clinical signs of experimental infection began after 4 days and continued for 10 days. The oocysts appeared in IFAT as spherical in shape with clear walls and bright yellowish green color. There was no significant difference noticed between the 2 methods of diagnosis [stain and immunological test]

Immunization against the colonization factor antigen I of enterotoxigenic Escherichia coli by administration of a bivalent Salmonella typhimurium aroA strain

An expression plasmid (pCFA-1) carrying the cfaB gene that codes for the enterotoxigenic Escherichia coli (ETEC) fimbrial adhesin colonization factor antigen I(CFA/I) subunit was constructed and used to transform a derivative of the attenuated Salmonella typhimurium aroA vaccine strain SL3261 carrying an F'lacl(q). Treatment of the transformed strain with isopropyl-beta-D-thiogalactopyranoside (IPTG) resulted in elevated in vitro expression of the CFA/I subunit. Although flagellar function and lipopolysaccharide (LPS) synthesis were similar in both the parental and the recombinant strains, spleen colonization was reduced in the recombinant strain. AII BALB/c mice parenteally inoculated with the recombinant strain developed significant anti-CFA/I and anti-LPS serum antibody titers (P<0.05). Moreover, 2 of 5 mice orally inoculated with the engineered Salmonella strain developed anti-CFA/I intestinal IgA (P>0.05) while 4/5 of the same mice developed anti-LPS (P<0.05). The results indicate that the vaccine strain elicited an antibody response against the bacterial host both after oral and intravenous immunization while the response against the CFA/I antigen was significant only after inoculation by the intravenous route.

Diarrea crónica y Cryptosporidium en pacientes diabéticos con subpoblación linfocitaria normal: informe de dos casos / Chronic diarrhea and Cryptosporidium in diabetic patients with normal lymphocyte subset counts: report of two cases

Se describen dos casos de pacientes diabéticos con cuentas normales de linfocitos TCD4+, que presentaban diarrea crónica y en los cuales se detectó Cryptosporidium en heces. En ambos casos las pruebas serológicas para VIH resultaron negativas. El hecho de que estos pacientes desarrollaran una patología que se observa comunmente en presencia de cuentas bajas de linfocitos TCD4+, sugiere que una alteración inmunológica distinta de la celular pudiera estar involucrada en la patogénesis de esta infección. Los autores sugieren que la búsqueda intencionada de Cryptosporidium debe considerarse en el estudio de la diarrea crónica del paciente diabético

Assessment of the immune and nutritional status of the host in childhood diarrhoea due to cryptosporidium.

Cryptosporidium oocysts were detected microscopically in the concentrated faecal smears (stained by modified kinyoun's acid fast stain) in 13 out of 100 (13 per cent) cases of acute diarrhoea (AD < 2 weeks duration), 7 out of 50 (14 per cent) cases of chronic diarrhoea (CD > 2 weeks duration) and none in 50 age matched controls. The grades of malnutrition of the cases and controls were calculated by the weight for age criteria and the immune status assessed by the levels of serum immunoglobulins and SIgA in duodenal fluids. Malnutrition was observed in 6 out of 13 cases (46.1 per cent) in acute and 6 out of 7 cases (85.71 per cent) in chronic cryptosporidial diarrhoeas. There was no significant statistical difference (P > 0.05) in serum immunoglobulins and SIgA levels in chronic cryptosporidiosis. SIgA was significantly reduced (P > 0.05) in cases of acute cryptosporidiosis. Cryptosporidium is an important cause of symptomatic infection in apparently immunocompetent children not having been detected in a single non-diarrhoeal control. Further a low SIgA could contribute to acute symptomatic cryptosporidiosis by favouring colonization with the parasite.

Flora autóctona intestinal: Revisión Bibliográfica / Autochthonous intestinal flora: bibliographic review

Se revisan diversos aspectos sobre la flora autóctona intestinal en el niño sano y en el enfermo. Su asociación con la antibioticoterapia y la dieta, los efectos metabólicos sobre la flora intestinal, su manipulación y su relación con el sistema inmunológico

Isospora belli in immunosuppressed patients

Two hundred diarrhoeic immunosuppressed patients were chosen from the outpatient, inpatient and oncology department of Zagazig University Hospital. Stool samples were taken and concentrated by formol ether and Sheather's sucrose techniques to detect Isospora and other protozoal parasites. Differentiation of the collected protozoal parasites were done by using modified acid fast, Giemsa and Iron haematoxylin stains. Isospora belli was found alone in 13 patients [6.5%] and associated with other protozoa in 10 patients [5%]. The modified acid fast stain after formal in ether concentration was the best method for differentiation of Isospora belli oocyst in stool. In association with diarrhoea, abdominal pain, malaise and flatulence were the main clinical presentation in infected patient. Diarrhoea was watery, chronic [up to 6 months] and of moderate degree [6 - 10 motions/day] in patients aged 20 - 40 years. Mucus and charcot leyden crystals were the main stool characters. For the first time, IFAT was done by using antigen from the pure strain of Eimeria stiedae, [rabbit hepatic coccidiosis], for serological diagnosis of Isospora belli. The specificity of the test was 100% and the sensitivity was 69%

Prioridades en la investigación de vacunas contra las enfermedades diarreicas / Research priorities for diarrheal disease vaccines: Memorandum from a WHO Meeting

Las diarreas causadas por rotavirus, Shigella, Vibrio cholera y Escherichia coli enterotoxigena representan una carga importante para los servicios de salud en los países en desarrollo y han estimulado numerosas actividades orientadas a preparar vacunas contra las enfermedades producidas por estos cuatro organismos patogenos. Este memorandum describe los adelantos mas recientes y senala el camino para futuras investigaciónes y ensayos clinicos en esta área

Diarreas infecciosas / Infections Diarrhea

Los autores de esta obra conscientes del problema de esta patología como problema de salud pública, trataron de organizar el material bibliográfico disponible a nivel nacional e internacional para incluir los temas en este texto, el cual es accesible a especialistas, medicos generales, estudiantes de medicina y publico en general

Vacunas contra Escherichia coli causante de diarrea en humanos / Vaccines against scherichia coli causing of diarrhea in humans

En este capítulo se describen las interacciones entre diferentes tipos de cepas de Escherichia coli con las células del epitelio intestinal, lo cual lleva a que el hospedero presente diarrea. El conocimiento de estas interacciones, emanado de veinte años de investigación, han permitido en fechas recientes plantear el uso de vacunas como medida de control de la enfermedad diarreica. La asociación constante entre los serotipos E. coli con procesos diarreicos, ha permitido plantear medidas de control a través de la búsqueda de los mecanismos comunes de patogenicidad que comparten la mayoría de los serotipos de E. coli. Estos mecanismos corresponden fundamentalmente a tres tipos: 1) adhesivos, que permiten a las bacterias acercarse, pegarse y colonizar el epitelio de ciertas áreas del intestino; 2) producción de proteínas bacterianas (toxinas) que estimulan la secreción de agua y electrolitos al interactuar con mecanismos bioquímicos de las células del huésped y 3) invasión y reproducción dentro del citoplasma de células epiteliales del intestino para evadir los mecanismos de protección del hospedero

Estudo em camundongos (CBA x C57B1/10) F1 da influência da enterotoxina termoestável (STa) de Escherichia coli na fagocitose / Study in mice (CBA x C57B1/10) F1 of the influence of termostable (STa) enterotoxin from Escherichia coli in phagocytosis

Camundongos (CBA x C57B1/10) F1 foram utilizados para se estudar a atividade fagocitária de seus macrófagos após a inoculaçäo, nos animais, da enterotoxina termoestável (STa) de Escherichia coli, obtida a partir de um amostra origem humana (sorogrupo 0128), semipurificada de acordo com Ricci e col (1982). Foi utilizada a técnica de Depuraçäo de Carbono Coloidal em camundongos inoculados com 250 unidades tóxicas (UT) de STa, em diferentes intervalos de tempo. Os resultados mostraram que 25OUT de STa aumentaram a velocidade de depuraçäo do carbono coloidal quando inoculados com a toxina 24, 48, 72 e 96 horas antes da administraçäo da substância coloidal. Em incoulaçöes feitas antes e após estes tempos näo houve interferência signficativa da toxina, sendo que a injeçäo de STa 48 horas antes do carbono coloidal, correspondia ao pico de depuraçäo do mesmo