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Id: biblio-970491
Autor: Ferreira, M. R. A; Stella, A. E; Freitas-Filho, E. G; Silva, T. S; Nascimento, K. A; Pinto, J. F. N; Dias, M; Moreira, C. N.
Título: Distribution of the stx1 and stx2 genes in Escherichia coli isolated from milk cattle according to season, age, and production scale in southwestern region of Goiás, Brazil / Distribuição dos genes stx1 e stx2 em Escherichia coli isoladas de bovinos de leite de acordo com a estação, idade e escala de produção na região sudoeste de Goiás, Brasil
Fonte: Arq. bras. med. vet. zootec. (Online);70(6):1807-1813, nov.-dez. 2018. tab.
Idioma: en.
Resumo: This study determined the distribution of stx1 and stx2 genes in Escherichia coli isolated from dairy herds with regard to animal age, season, and farm production-scale, and analyzed the phylogenetic distribution of the groups A, B1, B2, and D of 276 isolates of bovine feces Shiga toxin-producing E. coli (STEC). The stx1 profile was the most common, detected in 20.4% (202/990) of the isolates, followed by stx2 (4.54%, 45/990) and stx1+stx2 (2.92%, 29/990). The stx1 gene was detected more frequently in calves than in adult animals. In the dry season (winter), the presence of stx1+stx2 profile in cattle feces was higher than in the rainy season (summer), while no significant changes were observed between seasons for the stx1 and stx2 profiles. The most predominant phylogenetic groups in adult animals were B1, A, and D, while groups A and B1 prevailed in calves. Our data highlight the importance of identifying STEC reservoirs, since 7.5% of the tested isolates were positive for stx2, the main profile responsible for the hemolytic-uremic syndrome (HUS). Moreover, these microorganisms are adapted to survive even in hostile environments and can contaminate the food production chain, posing a significant risk to consumers of animal products.(AU)

Esse estudo determinou a distribuição dos genes stx1 e stx2 em Escherichia coli isolados de rebanhos leiteiros em relação a idade, estação e produção, e analisaram a distribuição filogenética dos grupos A, B1, B2 e D de 276 E. coli produtoras de toxina Shiga (STEC). O perfil stx1 foi mais comum, detectado em 20,4% (202/990) dos isolados, seguido de stx2 (4,54%, 45/990) e stx1+stx2 (2,92%, 29/990). O gene stx1 foi detectado mais frequentemente em bezerros que animais adultos. No período de seca (inverno), a presença do perfil stx1+stx2 nas fezes dos bovinos foi mais prevalente que no período chuvoso (verão), apesar de não haver diferença significativa entre estações para os perfis stx1 e stx2. Os grupos filogenéticos mais predominantes em animais adultos foram B1, A e D, enquanto grupos A e B2 prevaleceram em bezerros. Nossos dados enfatizam a importância de se detectar reservatórios de STEC já que 7,5% dos isolados testados foram positivos para stx2, o perfil mais prevalente em casos de síndrome hemolítica-urêmica. Ademais, esses microorganismos são adaptados à sobreviver em ambientes hostis e contaminam a cadeia alimentar, levando a risco significativo para consumidores de alimentos de origem animal.(AU)
Descritores: Bovinos/genética
Toxina Shiga I/genética
Toxina Shiga II/genética
Escherichia coli/isolamento & purificação
Escherichia coli/genética
Limites: Animais
Bovinos
Responsável: BR68.1 - Biblioteca Virginie Buff D'Ápice


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Id: lil-751888
Autor: Amani, Jafar; Ahmadpour, Askary; Fooladi, Abbas Ali Imani; Nazarian, Shahram.
Título: Detection of E. coli O157:H7 and Shigella dysenteriae toxins in clinical samples by PCR-ELISA
Fonte: Braz. j. infect. dis;19(3):278-284, May-Jun/2015. tab, graf.
Idioma: en.
Resumo: Shiga toxin producing bacteria are potential causes of serious human disease such as hemorrhagic colitis, severe inflammations of ileocolonic regions of gastrointestinal tract, thrombocytopenia, septicemia, malignant disorders in urinary ducts, hemolytic uremic syndrome (HUS) Shiga toxin 1 (stx1), shiga toxin 2 (stx2), or a combination of both are responsible for most clinical symptoms of these diseases. A lot of methods have been developed so far to detect shiga toxins such as cell culture, ELISA, and RFPLA, but due to high costs and labor time in addition to low sensitivity, they have not received much attention. In this study, PCR-ELISA method was used to detect genes encoding shiga toxins 1 and 2 (stx1 and stx2). To detect stx1 and stx2 genes, two primer pairs were designed for Multiplex-PCR then PCR-ELISA. PCR products (490 and 275, respectively) were subsequently verified by sequencing. Sensitivity and specificity of PCR-ELISA method were determined by using genome serial dilution and Enterobacteria strains. PCR-ELISA method used in this study proved to be a rapid and precise approach to detect different types of shiga toxins and can be used to detect bacterial genes encoding shiga toxins.
Descritores: /química
ESCHERICHIA COLI OACIDULATED PHOSPHATE FLUORIDE/química
Toxina Shiga I/isolamento & purificação
/isolamento & purificação
SHIGA TOXIN TEMEFOS/isolamento & purificação
Shigella dysenteriae/química
-DNA Bacteriano/genética
Ensaio de Imunoadsorção Enzimática
/genética
ESCHERICHIA COLI OACIDULATED PHOSPHATE FLUORIDE/genética
Fezes/microbiologia
Genes Bacterianos/genética
Reação em Cadeia da Polimerase
Sensibilidade e Especificidade
Toxina Shiga I/genética
/genética
SHIGA TOXIN TEMEFOS/genética
Shigella dysenteriae/genética
Limites: Adulto
Idoso
Criança
Feminino
Humanos
Masculino
Pessoa de Meia-Idade
Tipo de Publ: Research Support, Non-U.S. Gov't
Responsável: BR1.1 - BIREME


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Id: lil-634501
Autor: Gomez, D.; Miliwebsky, E.; Silva, A.; Deza, N.; Zotta, C.; Cotella, O.; Martínez Espinosa, E.; Chinen, I.; Fernández Pascua, C.; Rivas, M..
Título: Aislamiento de Escherichia coli productor de toxina Shiga durante un brote de gastroenteritis en un Jardín Maternal de la Ciudad de Mar del Plata / Isolation of Shiga toxin-producing Escherichia coli strains during a gastrointestinal outbreak at a day care center in Mar del Plata City
Fonte: Rev. argent. microbiol;37(4):176-183, oct.-dic. 2005. ilus, graf, tab.
Idioma: es.
Resumo: Entre el 15 de octubre y el 8 de noviembre de 2003 ocurrió un brote de gastroenteritis en un Jardín Maternal de un Hospital de la ciudad de Mar del Plata. Catorce de un total de 80 niños (17,5%), edad promedio 23,6 ± 13,9 meses, presentaron diarrea, y un caso evolucionó a síndrome urémico hemolítico. La madre de uno de los afectados presentó diarrea simultáneamente. No se pudo establecer el origen del brote, pero probablemente la transmisión haya sido fundamentalmente persona a persona. Las prácticas habituales en el lactario del jardín maternal, y las condiciones inadecuadas de infraestructura y hábitos de higiene de la cocina del Hospital fueron señalados como factores de riesgo. En un caso se detectó Escherichia coli productor de toxina Shiga (STEC) O103:H2, y STEC O26:H11 en otro. En el niño infectado por STEC O26:H11, la excreción se extendió por un período de 37 días. La no detección de STEC en aquellos casos en los cuales el intervalo entre el inicio de los síntomas y la toma de muestra fue mayor a 6 días, enfatiza la necesidad de la recolección temprana de especímenes. Las principales conclusiones de este estudio fueron la necesidad de establecer normas óptimas de higiene, informar rápidamente la ocurrencia de casos de gastroenteritis y confirmar la negativización de la excreción del patógeno.

From October 15 to November 8, 2003, a gastrointestinal outbreak occurred at a day care center in a Hospital in Mar del Plata City. Fourteen out of 80 (17.5%) children, mean age 23.6 ± 13.9 months, and the mother of one of them had diarrhea. One case developed hemolytic uremic syndrome. No conclusive evidence of the origin of the outbreak was found, but the epidemic curve suggested person-to-person spread. The usual practices at the place where infant milk formula was prepared at the day care center, together with the inadequate infrastructure conditions and hygiene practices at the kitchen of the hospital, were considered risk factors. One case had Shiga toxin-producing Escherichia coli (STEC) O103:H2 infection and other STEC O26:H11.The duration of shedding for the child with O26:H11 infection was 37 days. In the other symptomatic children, the pathogen was not recovered from fecal samples collected 6 or more days after the onset of the illness. This emphasizes that the collection of early samples is necessary to recover STEC strains. In order to prevent and control enteric diseases in day care facilities the following measures are necessary: optimal hygiene standards, early case reporting, and exclusion of those who remain culture-positive.
Descritores: Creches
Surtos de Doenças
Diarreia/microbiologia
Infecções por Escherichia coli/microbiologia
Escherichia coli/isolamento & purificação
Toxina Shiga I/análise
/análise
SHIGA TOXIN TEMEFOS/análise
-Argentina/epidemiologia
Diarreia Infantil/epidemiologia
Diarreia Infantil/microbiologia
Diarreia/epidemiologia
Infecções por Escherichia coli/epidemiologia
Infecções por Escherichia coli/transmissão
Escherichia coli/classificação
Escherichia coli/metabolismo
Síndrome Hemolítico-Urêmica/microbiologia
Fatores de Risco
Sorotipagem
Limites: Adulto
Pré-Escolar
Feminino
Humanos
Lactente
Masculino
Responsável: AR1.2 - Instituto de Investigaciónes Epidemiológicas


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Id: lil-634483
Autor: Leotta, G.A.; Chinen, I.; Epszteyn, S.; Miliwebsky, E.; Melamed, I.C.; Motter, M.; Ferrer, M.; Marey, E.; Rivas, M..
Título: Validación de una técnica de PCR múltiple para la detección de Escherichia coli productor de toxina Shiga / Validation of a multiplex PCR for detection of Shiga toxin-producing Escherichia coli
Fonte: Rev. argent. microbiol;37(1):1-10, ene.-mar. 2005. ilus, tab.
Idioma: es.
Projeto: Consejo Nacional de Investigaciones Científicas y Tecnológicas. PIP Nº 0020/98.
Resumo: La infección por Escherichia coli productor de toxina Shiga (STEC) es causa de diarrea con o sin sangre, colitis hemorrágica y síndrome urémico hemolítico (SUH) en humanos. El objetivo de este trabajo fue validar una técnica de PCR múltiple para el diagnóstico de STEC basado en la detección de los genes stx1, stx2 y rfbO157. La validación de la técnica se realizó en dos laboratorios independientes, en forma paralela. Se determinó rango de trabajo, selectividad y robustez. Se evaluó el desempeño de la técnica al combinar distintas concentraciones de dos cepas con diferentes factores de virulencia. El rango de trabajo dependió de la cepa analizada, los valores máximos y mínimos fueron 6,6 x 107 y 1,0 x 104 UFC/50 µl. El límite de detección fue de 1,0 x 104 UFC/50 µl y el límite de corte de 1,0 x 105 UFC/50 µl. La robustez fue óptima al modificar diferentes variables. Se obtuvo 100% de inclusividad, exclusividad, precisión analítica, valor predictivo positivo y valor predictivo negativo. No se observó interferencia al combinar distintas concentraciones de los factores de virulencia blanco de la reacción. La técnica validada es una alternativa apropiada para la detección y confirmación de STEC O157 y no-O157 a partir de cultivos bacterianos.

Shiga toxin-producing Escherichia coli (STEC) cause non-bloody or bloody diarrhea, hemorrhagic colitis and hemolytic uremic syndrome (HUS) in humans. The aim of the present study was to validate a multiplex PCR for the STEC diagnosis based on the detection of stx1, stx2 and rfbO157 genes. The multiplex PCR validation was carried out in two independent laboratories in a parallel way. Work range, selectivity and robustness were established. The PCR performance was evaluated using different concentrations of two STEC strains harboring different target genes. The work range depended on the strain analyzed, the maximum and the minimum values were 6.6 x 107 and 1.0 x 104 CFU/50 µl. The detection limit was 1.0 x 104 CFU/50 µl and the cut limit 1.0 x 105 CFU/50 ml. A good robustness was observed when different variables were introduced. Inclusivity, exclusivity, positive predictivity, negative predictivity and analytical accuracy were of 100%. Interference was not shown when different concentrations of STEC strains, carrying different genes, were used. The validated technique is an appropriate alternative for detection and confirmation of STEC O157 and non-O157 strains from bacterial cultures.
Descritores: Escherichia coli/genética
Reação em Cadeia da Polimerase/métodos
Toxinas Shiga/genética
-Fracionamento Celular/instrumentação
Detergentes
DNA Bacteriano/genética
DNA Bacteriano/isolamento & purificação
Escherichia coli/química
Escherichia coli/classificação
Polietilenoglicóis
Especificidade da Espécie
Toxina Shiga I/genética
/genética
SHIGA TOXIN TEMEFOS/genética
Tipo de Publ: Estudo Comparativo
Research Support, Non-U.S. Gov't
Estudo de Validação
Responsável: AR1.2 - Instituto de Investigaciónes Epidemiológicas


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Id: lil-595679
Autor: Bouzari, Saeid; Aslani, Mohammad M; Oloomi, Mana; Jafari, Anis; Dashti, Amir.
Título: Comparison of multiplex PCR with serogrouping and PCR-RFLP of fliC gene for the detection of enteropathogenic Escherichia coli (EPEC)
Fonte: Braz. j. infect. dis;15(4):365-369, July-Aug. 2011. ilus, tab.
Idioma: en.
Projeto: Pasteur Institute of Iran.
Resumo: Enteropathogenic Escherichia coli (EPEC) comprise one of the six categories of diarrhoeagenic E. coli (DEC). EPEC is subgrouped into typical (tEPEC) and atypical (aEPEC). The identification of DEC cannot be based only on cultural and biochemical criteria, since they are indistinguishable from the non-pathogenic E. coli commonly found in human feces. Several PCR methods, with both single and multiple target genes, have been reported for detecting the different DEC pathotypes. In the present study five hundred E. coli isolates from children with diarrhea were subjected into multiplex PCR. Furthermore the strains were typed serologically with O antisera and their fliC gene was characterized by PCR-RFLP. The results obtained revealed that overall 41 (8.2 percent) isolates could be detected as EPEC by this multiplex PCR assay. Of these isolates; 27 (66 percent) were typical (escv+, bfp+) and 14 (34 percent) atypical EPEC (escv+, bfp-). None of these 41 isolates contained the Stx1 and Stx2 genes. Among 37 (90 percent) typeable strains, nine different serogroups were present. The most common serogroups were O111, followed by O86, O55 and O119 and 10 different H types were found among these isolates. The multiplex PCR assay was found to be rapid and reliable in comparison to serological test; especially when screening the large number of isolates.
Descritores: DNA Bacteriano/análise
Escherichia coli Enteropatogênica/classificação
Proteínas de Escherichia coli/genética
Reação em Cadeia da Polimerase Multiplex
Antígenos O/análise
Polimorfismo de Fragmento de Restrição
-Diarreia/microbiologia
Escherichia coli Enteropatogênica/genética
Escherichia coli Enteropatogênica/isolamento & purificação
Fezes/microbiologia
Sorotipagem/métodos
Toxina Shiga I/genética
/genética
SHIGA TOXIN TEMEFOS/genética
Limites: Criança
Humanos
Tipo de Publ: Estudo Comparativo
Research Support, Non-U.S. Gov't
Responsável: BR1.1 - BIREME


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Id: lil-445766
Autor: Valles, P. G; Pesle, S; Piovano, L; Davila, E; Peralta, M; Principi, I; Lo Giudice, P.
Título: Postdiarrheal Shiga toxin-mediated hemolytic uremic syndrome similar to septic shock
Fonte: Medicina (B.Aires);65(5):395-401, 2005. tab.
Idioma: en.
Conferência: Apresentado em: Congreso Argentino de Nefrología Pediátrica, 3, Buenos Aires, 20-23 june 2003.
Resumo: The inflammatory response of host endothelial cells is included in the development of vascular damage observed in enterohemorrhagic Escherichia coli (EHEC) infection, resulting in hemolytic uremic syndrome (HUS). The response to a non-conventional treatment for a group of D+ HUS (diarrhea positive HUS) patients, with clinical hemodynamic parameters of septic shock was evaluated in this prospective study (1999-2003). Twelve children 2.8 +/- 0.6 years old, with D+ HUS produced by E. coli infection with serological evidence of Shiga toxin, presenting severe unstable hemodynamic parameters and neurological dysfunction at onset, were studied. The protocol included fresh frozen plasma infusions, methylprednisolone pulses (10mg/k/day) for three consecutive days and plasma exchange for five days, starting after admission to the intensive care unit (ICU). The twelve patients with increased pediatric risk of mortality (PRISM) score: 18 +/- 2 after admission to intensive care unit (ICU), required dialysis for 17.4 +/- 4 days, mechanical ventilator assistance for 10 +/- 1 days and early inotropic drugs support for 10.5 +/- 1 days. Neurological dysfunction included generalized tonic-clonic seizures lasting for 5.4 +/- 1 days, n:8. Focal seizures were present in the remaining patients. Dilated cardiomyopathy was present in 6 children. Eight children suffered hemorrhagic colitis. Nine patients survived. Within one year of the injury, neurological sequelae, Glasgow outcome scale (GOS) 3 and 4, were present in two patients, chronic renal failure in one patient. We suggest that early introduction of this protocol could benefit D+ HUS patients with hemodynamic instability and neurological dysfunction at onset. Further studies are likely to elucidate the mechanisms involved in this early adverse clinical presentation of D+ HUS patients.

La respuesta inflamatoria de la célula endotelial se incluye en el desarrollo del daño vascular observado en la infección por Escherichia coli enterohemorrágica que deviene en Síndrome Urémico Hemolítico (SUH). Se evaluó en forma prospectiva, entre 1999 y 2003, la respuesta a un tratamiento no convencional, en doce pacientes, edad 2.8 ± 0.6 años, que desarrollaron SUH con presencia de diarrea sanguinolenta (SUH D+) y evidencia serológica de toxina Shiga, los cuales en fase inicial presentaron parámetros hemodinámicoscompatibles con shock séptico y compromiso neurológico grave. El protocolo incluyó transfusión de plasmafresco, pulsos de metilprednisolona (10mg/k/día) por tres días consecutivos y plasmaféresis por cinco días, iniciados en las primeras 48 horas. Los doce pacientes ingresaron en terapia intensiva, presentando unapuntuación de riesgo de mortalidad pediátrica (PRISM): 18 ± 2, con requerimiento de diálisis por 17.4 ± 4 días, asistencia ventilatoria mecánica por 10 ± 1días y soporte temprano con drogas inotrópicas por un período de10.5 ± 1 días. La disfunción neurológica se presentó con convulsiones tónico-clónicas generalizadas por 5.4 ±1 días en 8 pacientes y con convulsiones focalizadas en los restantes. Seis pacientes desarrollaron miocardiopatíadilatada y 8 presentaron colitis hemorrágica. Sobrevivieron a la etapa aguda de la enfermedad 9 pacientes. Alfinalizar el primer año de seguimiento, dos de ellos presentaban secuelas neurológicas (escala de seguimientode Glasgow; GOS 3 y 4 respectivamente) y uno, fallo renal crónico. La introducción temprana de este protocolo podría beneficiar a pacientes con SUH D+ con inestabilidad hemodinámica grave y disfunción neurológica al inicio. Los mecanismos involucrados en esta temprana presentación clínica adversa de SUH D+ permanecen aún sin dilucidar.
Descritores: Choque Séptico/fisiopatologia
Diarreia/fisiopatologia
Infecções por Escherichia coli/fisiopatologia
Síndrome Hemolítico-Urêmica/fisiopatologia
-Diarreia/complicações
Diarreia/terapia
/isolamento & purificação
ESCHERICHIA COLI OACIDULATED PHOSPHATE FLUORIDE/isolamento & purificação
Infecções por Escherichia coli/complicações
Infecções por Escherichia coli/terapia
Reação em Cadeia da Polimerase
Estudos Prospectivos
Toxina Shiga I
Toxina Shiga II
Estatísticas não Paramétricas
Síndrome Hemolítico-Urêmica/microbiologia
Síndrome Hemolítico-Urêmica/terapia
Resultado do Tratamento
Limites: Criança
Pré-Escolar
Humanos
Lactente
Responsável: BR1.1 - BIREME


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Id: lil-333517
Autor: Ramírez, Margarita; Morier, Luis; Alonso, Janette; Bravo, Laura; Cabrera, Roberto; Fernández, Anabel.
Título: Determinación de verotoxinas en cepas de Escherichia coli O157: H7 / Determination of verotoxins in strains of Escherichia coli O157: H7
Fonte: Rev. cuba. med. trop;51(3):152-155, Sept.-Dec. 1999.
Idioma: es.
Resumo: We performed an study to find out the main virulence factor in verotoxigenic Escherichia coli: the production of verotoxins in 50 non sorbitol-fermenting Escherichia coli strains (possible enterohemorrhage) which were isolated from children with acute diarrheas in the City of Havana and referred to the National Reference Laboratory for acute diarrheal diseases in "Pedro KourÝ" Institute. By using the agglutination technique with latex particles of E. coli O157:H7, we determined whether the verotoxins belonged to this serotype, we also researched the production of verotoxins in Vero cell culture. Ninety-six percent of the total number of strains were positive in the qualitative determination of this factor which was more frequently observed after 24 hours.
Descritores: Toxinas Bacterianas
Escherichia coli O157
-Toxina Shiga I
Responsável: BR1.1 - BIREME


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Id: lil-331803
Autor: Gómez, D.
Título: Aislamiento y caracterización de Escherichia coli productor de toxina Shiga en hamburguesas supercongeladas y quesos de pasta blanda / Isolation and characterization of Shiga-toxin-producing Escherichia coli from frozen hamburgers and soft cheeses
Fonte: Rev. argent. microbiol;34(2):66-71, abr.-jun. 2002.
Idioma: es.
Resumo: Shiga toxin producing-Escherichia coli (STEC), an important emerging foodborne pathogen, has been associated with bloody and non-bloody diarrhea, hemorrhagic colitis, hemolytic uremic syndrome (HUS) and thrombotic thrombocytopenic purpura. The cattle have been shown to be a major reservoir of STEC and raw foods such as ground beef and milk are the most common vehicles of infection. In the present study, the prevalence of STEC in 95 samples of frozen hamburgers and in 114 samples of soft cheese was established in 8.4 and 0.9, respectively. The genotypic and phenotypic characteristics of the strains were determined. The virulence genes stx1, stx2, eaeA and EHEC-hlyA were identified by PCR and by colony blot hybridization assays. Serotyping, antimicrobial susceptibility and production of Stx using specific cytotoxicity assays on Vero cells were also determined. All STEC strains were characterized as eaeA-/EHEC-hlyA+. The stx2 genotype was prevalent (77.8), and four different O:H serotypes were found, comprising: O8:H19 (5 strains), O113:H21 (1), O8:H16 (1), and O39:H49 (1). One STEC strain was nontypable. Although soft cheese complimented the microbiological quality controls for the coliform counts, the detection of STEC in one sample raises doubts concerning the effectiveness of the current quality controls. These data contribute to the implementation of strategies for the prevention and control of HUS.
Descritores: Queijo
Escherichia coli
Contaminação de Alimentos
Microbiologia de Alimentos
Carne
Toxina Shiga I
Toxina Shiga II
-Adesinas Bacterianas
Argentina
Toxinas Bacterianas
Chlorocebus aethiops
Criopreservação
Escherichia coli
Inspeção de Alimentos
Conservação de Alimentos
Genótipo
Proteínas Hemolisinas
Fenótipo
Proteínas de Escherichia coli/biossíntese
Proteínas de Transporte/biossíntese
Proteínas de Transporte/genética
Resistência a Medicamentos/genética
Toxina Shiga I
Toxina Shiga II
Células Vero
Virulência
Limites: Animais
Bovinos
Tipo de Publ: Research Support, Non-U.S. Gov't
Responsável: BR1.1 - BIREME



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