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1.
Vet Microbiol ; 105(1): 29-36, 2005 Jan 05.
Artigo em Inglês | MEDLINE | ID: mdl-15607081

RESUMO

In order to determine the occurrence, serotypes and virulence markers of Shiga toxin-producing Escherichia coli (STEC) strains, 153 fecal samples of cattle randomly selected from six dairy farms in Sao Paulo State, Brazil, were examined for Shiga toxin (Stx) production by the Vero cell assay. Feces were directly streaked onto MacConkey Sorbitol Agar and incubated at 37 degrees C overnight. Sorbitol-negative colonies (maximum 20) and up to 10 sorbitol-positive colonies from each plate were subcultured onto presumptive diagnostic medium IAL. Sorbitol-negative isolates were screened with O157 antiserum for identification of O157:H7 E. coli. Isolates presenting cytotoxic activity were submitted to colony hybridization assays with specific DNA probes for stx1, stx2, eae, Ehly and astA genes. The isolation rate of STEC ranged from 3.8 to 84.6% depending on the farm analysed. STEC was identified in 25.5% of the animals, and most of them (64.1%) carried a single STEC serotype. A total of 202 STEC isolates were recovered from the animals, and except for the 2 O157:H7 isolates all the others expressed cytotoxic activity. The great majority of the STEC isolates carried both stx1 and stx2 genes (114/202, 56.4%) or stx2 (82/202, 40.6%); and whereas the Ehly sequence occurred in most of them (88%) eae was only observed in O157:H7 and O111:HNM isolates. Serotypes O113:H21, O178:H19 and O79:H14 were the most frequent STEC serotypes identified and widely distributed among animals from different farms, while others such as O77:H18, O88:H25 and O98:H17 occurred only in particular farms. This is the first report on the occurrence of STEC in dairy cattle in Sao Paulo State, and the results point to substantial differences in rate of isolation, serotypes and genetic profile of STEC that has been previously described among beef cattle in our community. Moreover, to our knowledge O79:H14 and O98:H17 represent new STEC serotypes, while O178:H19 has only been recently reported in Spain.


Assuntos
Doenças dos Bovinos/microbiologia , Infecções por Escherichia coli/veterinária , Escherichia coli O157/classificação , Toxina Shiga I/biossíntese , Toxina Shiga II/biossíntese , Adesinas Bacterianas/genética , Adesinas Bacterianas/metabolismo , Animais , Toxinas Bacterianas/genética , Toxinas Bacterianas/metabolismo , Brasil , Bovinos , DNA Bacteriano/química , DNA Bacteriano/genética , Indústria de Laticínios , Infecções por Escherichia coli/microbiologia , Escherichia coli O157/genética , Escherichia coli O157/crescimento & desenvolvimento , Escherichia coli O157/patogenicidade , Proteínas de Escherichia coli/genética , Proteínas de Escherichia coli/metabolismo , Fezes/microbiologia , Feminino , Hibridização de Ácido Nucleico , Antígenos O/metabolismo , Sorotipagem , Toxina Shiga I/genética , Toxina Shiga II/genética
2.
Clin Infect Dis ; 38(11): 1545-51, 2004 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-15156440

RESUMO

Diarrheal diseases are a leading cause of childhood morbidity and mortality in Latin America. Most studies have focused on infants but not on older children. We enrolled 505 children (age, 12-59 months) with diarrhea and age-matched controls in a case-control study in Sao Paulo, Brazil. Independent risk factors for diarrhea included another household member with diarrhea (matched odds ratio [mOR], 8.1; attributable fraction [AF], 0.17; P<.001) and consumption of homemade juice (mOR, 1.8; AF, 0.10; P=.01); protective factors included boiling of the baby bottle or nipple (mOR, 0.60; AF, 0.19; P=.026), childcare at home (mOR, 0.58; AF, 0.12; P=.004), and piped sewage (mOR, 0.58; AF, 0.05; P=.047). Hand washing by the caretaker after helping the child defecate protected against Shigella infection (mOR, 0.35; P<.05). Preparation of rice, beans, or soup in the morning and serving it to children after noon were associated with enterotoxigenic Escherichia coli infection (mOR, 8.0; P<.05). In these poor households, 28% of cases of diarrhea in 1-4-year-old children was attributable to easily modifiable exposures.


Assuntos
Diarreia Infantil/epidemiologia , Diarreia/epidemiologia , Infecções por Adenovirus Humanos/epidemiologia , Infecções por Adenovirus Humanos/prevenção & controle , Idoso , Brasil/epidemiologia , Infecções por Campylobacter/epidemiologia , Infecções por Campylobacter/prevenção & controle , Estudos de Casos e Controles , Pré-Escolar , Diarreia/microbiologia , Diarreia/prevenção & controle , Diarreia/virologia , Diarreia Infantil/microbiologia , Diarreia Infantil/prevenção & controle , Diarreia Infantil/virologia , Disenteria Bacilar/epidemiologia , Disenteria Bacilar/prevenção & controle , Infecções por Escherichia coli/epidemiologia , Infecções por Escherichia coli/prevenção & controle , Fezes/microbiologia , Fezes/virologia , Feminino , Humanos , Lactente , Masculino , Seleção de Pacientes , Valor Preditivo dos Testes , Fatores de Risco , Infecções por Rotavirus/epidemiologia , Infecções por Rotavirus/prevenção & controle , Distribuição por Sexo , Shigella/isolamento & purificação , Shigella/patogenicidade , Especificidade da Espécie
3.
Biomaterials ; 25(2): 327-33, 2004 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-14585720

RESUMO

The aim of this study was to evaluate the physical-chemical properties, in vivo biocompatibility and antimicrobial activity of bioactive glasses (BG) used as a controlled release device for tetracycline hydrochloride and an inclusion complex formed by tetracycline and beta-cyclodextrin at 1:1 molar ratio. The BG as well as their compounds loaded with tetracycline (BT) and tetracycline:beta-cyclodextrin (BTC) were characterized by FTIR spectroscopy, X-ray powder diffraction, differential scanning calorimetry and by scanning electron microscopy and energy dispersive spectroscopy. The in vivo test was carried out with female mice split into three groups treated with bioactive glass either without drugs, or associated with tetracycline, or with tetracycline:beta-cyclodextrin by subcutaneous implantation. The histological examination of tissue at the site of implantation showed moderate inflammatory reactions in all groups after 72 h. The bacterial effect was tested on A. actinomycetemcomitans suspended in BHI broth, with or without bioactive particles. A considerable bacteriostatic activity was found with BT and BTC glasses, as compared to plain glass. The presence of cyclodextrin was important to slow down the release of tetracycline for a long period of time and it was verified that the presence of tetracycline or its inclusion complex, tetracycline:beta-cyclodextrin, did not affect the bioactivity of the glass.


Assuntos
Ciclodextrinas/farmacocinética , Sistemas de Liberação de Medicamentos , Vidro , Tetraciclina/farmacocinética , beta-Ciclodextrinas , Animais , Materiais Biocompatíveis/química , Ciclodextrinas/administração & dosagem , Ciclodextrinas/química , Vidro/química , Temperatura Alta , Camundongos , Espectroscopia de Infravermelho com Transformada de Fourier , Tetraciclina/administração & dosagem , Tetraciclina/química
4.
FEMS Microbiol Lett ; 152(1): 95-100, 1997 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-9228775

RESUMO

Genotypic and phenotypic virulence markers of the different categories of diarrheagenic Escherichia coli were investigated in 76 strains of the enteropathogenic E. coli (EPEC) serogroup O125. The most frequent serotype found was O125ac:H21. None of the serotypes behaved as EPEC, i.e. carried the eaeA, bfpA, and EAF DNA sequences simultaneously and presented localized adherence to HeLa cells. All strains of O125ac:H6 were atypical EPEC since they carried eaeA only, and presented an indefinite pattern of adherence. All strains of O125ab:H9, O125ac:H9, O125?:H16, and O125ab:H21 and 79% of the O125ac:H21 strains were enteroaggregative E. coli, since they carried a specific DNA sequence and presented the typical aggregative adherence pattern.


Assuntos
Diarreia Infantil/microbiologia , Escherichia coli/classificação , Humanos , Lactente , Recém-Nascido , Sorotipagem , Virulência
5.
FEMS Microbiol Lett ; 203(2): 199-205, 2001 Sep 25.
Artigo em Inglês | MEDLINE | ID: mdl-11583848

RESUMO

The pathogenic mechanisms of enteroaggregative Escherichia coli (EAEC) are not well defined. We investigated the interaction of EAEC strain 236 (serotype O111:H12) with polarised Caco-2 and T84 human intestinal epithelial cells lines, and with human jejunal and colonic mucosa. Strain 236 adhered to both polarised cell lines and to both intestinal tissue types, but caused severe damage and was invasive only in T84 cells and colonic mucosa. In contrast, prototype EAEC strain 042, which also adhered to the cultured intestinal cell lines, did not adhere to or invade jejunal or colonic tissue. These observations suggest a heterogeneity of virulence properties within the EAEC category of diarrhoea-causing E. coli.


Assuntos
Colo/microbiologia , Escherichia coli/patogenicidade , Mucosa Intestinal/microbiologia , Aderência Bacteriana , Células CACO-2 , Linhagem Celular , Criança , Escherichia coli/classificação , Humanos , Jejuno/microbiologia , Microscopia Eletrônica , Sorotipagem , Virulência
6.
FEMS Microbiol Lett ; 201(2): 163-8, 2001 Jul 24.
Artigo em Inglês | MEDLINE | ID: mdl-11470356

RESUMO

The expression of surface structures and the presence of DNA sequences related to putative virulence factors were investigated in 22 enteroaggregative Escherichia coli strains (EAEC). Fimbria was the most frequent (72.7%) structure identified. Only strains hybridising with the EAEC DNA probe carried aggA, but one strain produced a similar but unrelated bundle-like structure. All probe-positive and 62.5% of the probe-negative strains carried the virulence genes tested; aspU and irp2 prevailed among the former strains. The EAEC probe-positive strains were more diverse, and some of these strains, which promoted cell detachment, also carried the hly and pap sequences, thus suggesting they might represent uropathogenic E. coli.


Assuntos
Proteínas da Membrana Bacteriana Externa/química , Sondas de DNA/genética , Infecções por Escherichia coli/microbiologia , Escherichia coli/química , Escherichia coli/patogenicidade , Variação Genética/genética , Adesinas Bacterianas/análise , Adesinas Bacterianas/química , Adesinas Bacterianas/genética , Proteínas da Membrana Bacteriana Externa/análise , Proteínas da Membrana Bacteriana Externa/genética , Pré-Escolar , Diarreia/microbiologia , Escherichia coli/classificação , Escherichia coli/genética , Fímbrias Bacterianas/genética , Fímbrias Bacterianas/ultraestrutura , Marcadores Genéticos/genética , Genótipo , Testes de Hemaglutinação , Humanos , Lactente , Recém-Nascido , Microscopia Eletrônica , Virulência/genética
7.
FEMS Microbiol Lett ; 200(1): 117-22, 2001 Jun 12.
Artigo em Inglês | MEDLINE | ID: mdl-11410359

RESUMO

Escherichia coli strains of non-EPEC serotypes that carry eae and lack the EAF and the Shiga toxin (stx) gene sequences have been found in acute diarrhea. Both the cell association and the cell entry of these strains in human intestinal epithelial cells were studied as a function of cell differentiation and polarization. The eae+/EAF-/stx- non-EPEC E. coli strains invaded undifferentiated Caco-2 cells more efficiently than differentiated cells. In contrast, prototype EPEC strain E2348/69 did not show significative differences from invasion rates of undifferentiated and differentiated cells. The uptake of these strains was greatly enhanced by pretreatment of differentiated Caco-2 cells with EGTA. These results suggest that the eae+/EAF-/stx- non-EPEC E. coli invasion of intestinal cells may be dependent on receptors expressed on the surface of undifferentiated cells and the basolateral pole of differentiated cells.


Assuntos
Adesinas Bacterianas , Proteínas de Transporte , Enterócitos/microbiologia , Proteínas de Escherichia coli , Escherichia coli/patogenicidade , Genes Bacterianos , Intestinos/microbiologia , Aderência Bacteriana , Proteínas da Membrana Bacteriana Externa/genética , Células CACO-2 , Diferenciação Celular , Polaridade Celular , Enterócitos/citologia , Enterócitos/ultraestrutura , Escherichia coli/genética , Escherichia coli/ultraestrutura , Humanos , Intestinos/citologia , Intestinos/ultraestrutura , Plasmídeos/genética , Sorotipagem , Toxina Shiga/genética
8.
FEMS Microbiol Lett ; 146(1): 123-8, 1997 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-8997716

RESUMO

Seventeen Escherichia coli O111:H12 strains isolated from the feces of children with acute diarrhea were studied with regard to their adherence properties and other virulence characteristics. All strains showed an aggregative adherence pattern to HEp-2 cells and agglutinated bovine and sheep red cells in the presence of mannose. These strains did not have gene sequences homologous to the aggregative adherence fimbria I gene and did not react with any of the DNA probes used to detect other virulence genes in enteropathogens. With one exception, the O111:H12 strains did not induce fluid accumulation in the rabbit ileal loop assay, although 16 of the strains had the enteroaggregative E. coli heat-stable enterotoxin 1 (EAST) gene sequences. A 60-70 MDa plasmid was present in 16 of the strains studied. We conclude that the O111:H12 serotype, one of the first E. coli identified in infantile diarrhea, belongs to the enteroaggregative E. coli category but the genes encoding its adherence phenotype are distinct from those previously described.


Assuntos
Escherichia coli/classificação , Escherichia coli/patogenicidade , Animais , Aderência Bacteriana/genética , Sequência de Bases , Bovinos , Linhagem Celular , Criança , Primers do DNA/genética , Diarreia/microbiologia , Escherichia coli/genética , Infecções por Escherichia coli/microbiologia , Fímbrias Bacterianas/genética , Genes Bacterianos , Humanos , Reação em Cadeia da Polimerase , Coelhos , Sorotipagem , Virulência/genética
9.
FEMS Microbiol Lett ; 204(1): 105-10, 2001 Oct 16.
Artigo em Inglês | MEDLINE | ID: mdl-11682187

RESUMO

We evaluated a biochemical assay based on the ability to metabolise beta-phenylpropionic acid (PPA) as a diagnostic aid in the identification of typical enteropathogenic Escherichia coli (EPEC) strains. A total of 1061 E. coli strains of serogroups O55, O111, and O119 were initially characterised regarding their H types (serotypes) and the presence of EPEC DNA sequences, eae, EAF, and bfpA. In case of the serogroup O111 strains, 84.6% carried the typical EPEC markers, and the great majority of those (98.1%) were PPA-positive. In contrast, only 0.9% of the serogroups O55 and O119 strains carrying the typical EPEC markers (53.6% and 75.4%, respectively) were PPA-positive. We conclude that the PPA test is a useful method to detect typical EPEC strains only among strains of the O111 serogroup.


Assuntos
Infecções por Escherichia coli/diagnóstico , Escherichia coli/classificação , Escherichia coli/metabolismo , Fenilpropionatos/metabolismo , Proteínas de Bactérias/genética , Técnicas de Tipagem Bacteriana/métodos , Criança , Pré-Escolar , Escherichia coli/genética , Escherichia coli/patogenicidade , Infecções por Escherichia coli/microbiologia , Humanos , Sorotipagem , Virulência/genética
10.
FEMS Microbiol Lett ; 190(2): 293-8, 2000 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-11034294

RESUMO

A total of 919 Escherichia coli isolates from 125 children with diarrhoea (cases) and 98 controls were assayed for adherence to HEp-2 cells. Localised adherence was found only in isolates from cases. Diffuse, aggregative (AA), chain-like adherence (CLA) and variants of the AA pattern were found in both cases and controls. The AA isolates were tested for gene sequences associated with enteroaggregative E. coli (EAEC). Only 25% of the isolates hybridised with the EAEC probe, and the aafA, astA and pet gene sequences were found in 7.9%, 44.7% and 7.9% of the isolates, respectively. The aggA gene was not found, although 7.9% were positive for aggC. The CLA isolates reacted with the EAEC probe (55.6%), and the aggC, astA and pet gene sequences were found in 66.7%, 33.3% and 11.1%, respectively. The aggR (55.6%), aspU (55.6%), shf (33.3%) and she (22.2%) genes were also found in CLA isolates.


Assuntos
Aderência Bacteriana/fisiologia , Diarreia/microbiologia , Infecções por Escherichia coli/microbiologia , Escherichia coli/fisiologia , Escherichia coli/patogenicidade , Adesinas de Escherichia coli/genética , Adesinas de Escherichia coli/metabolismo , Brasil/epidemiologia , Criança , Pré-Escolar , Diarreia/epidemiologia , Escherichia coli/genética , Escherichia coli/isolamento & purificação , Infecções por Escherichia coli/epidemiologia , Humanos , Prevalência , Células Tumorais Cultivadas
11.
J Med Microbiol ; 47(9): 781-90, 1998 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-9736160

RESUMO

Faeces from urban children < 2 years old with acute diarrhoeal illness and from non-diarrhoeal infants (controls) were examined for Escherichia coli and other enteropathogens. A total of 990 E. coli isolates from 100 patients and 50 controls was tested for enteropathogenic E. coli (EPEC) serotype (O:H), adherence to HEp-2 cells after incubation for 3 and 6 h, fluorescent actin staining (FAS), DNA hybridisation with EAF, eaeA, STh, STp and EAggEC probes and production of heat-labile enterotoxin (LT) and verocytotoxin (VT) with Y1 and Vero cells. EPEC were the most prevalent enteropathogens in patients (32.7%; and 14% in controls). Enteroinvasive E. coli (EIEC) and Vero cytotoxin-producing E. coli (VTEC) were not detected. The rate of isolation of enterotoxigenic E. coli (ETEC) was identical in both groups. Among the EPEC isolates the prevalent serotypes were O111:H2, O55:NM and O119:H6. Localised adherence (LA) was found significantly more frequently in isolates from patients (19.6%) than controls (2.1%). All LA-positive EPEC isolates were FAS+ and eaeA+, but only 75.2% of them hybridised with the EAF probe. Diffusely adhering E. coli (DAEC) and enteroaggregative E. coli (EAggEC) were found with equal frequency in patients and controls. Twenty-seven E. coli isolates were negative for EAF but positive for eaeA and FAS and produced LA in 6-h adherence tests. These EAF-/eaeA+ strains were the only putative enteropathogen identified in seven patients and were not found in controls. The ability of these strains to elicit ultrastructural cell alterations and cell-signalling events was evaluated in Caco-2 cells (human colon carcinoma cell line) by the gentamicin invasion assay and by transmission electron microscopy. The numbers of intracellular bacteria in cell invasion tests varied from 0.4% to 1.6% of the cell-associated bacteria after a 6-h incubation period. Tyrosine phosphorylation of host cell proteins was assessed in HEp-2 cells by immunofluorescence microscopy and all strains gave positive results. EAF-/eaeA+ E. coli strains express most of the virulence properties found among true EPEC strains and can be a relevant cause of infant diarrhoea in developing countries.


Assuntos
Diarreia Infantil/microbiologia , Infecções por Escherichia coli/microbiologia , Proteínas de Escherichia coli , Escherichia coli/patogenicidade , Doença Aguda , Testes de Aglutinação , Aderência Bacteriana , Toxinas Bacterianas/biossíntese , Brasil/epidemiologia , Células CACO-2 , Linhagem Celular , Citotoxinas/biossíntese , Sondas de DNA , Diarreia Infantil/epidemiologia , Enterotoxinas/biossíntese , Escherichia coli/classificação , Escherichia coli/isolamento & purificação , Infecções por Escherichia coli/epidemiologia , Fezes/microbiologia , Humanos , Incidência , Lactente , Hibridização de Ácido Nucleico , Prevalência , Sorotipagem , Toxina Shiga I , População Urbana , Virulência
12.
Int J Food Microbiol ; 12(4): 333-8, 1991 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-1677256

RESUMO

Escherichia coli strains were isolated from 96 food samples (32 milks, 4 dairy products, 36 raw meats, 7 meat products, 7 sandwiches and 10 ready-to-eat meals). A total of 306 colonies was submitted to hybridization assays with DNA probes for the following virulence factors: heat-labile toxins (LT-I and LT-II), heat-stable toxins (ST-h and ST-p). Shiga-like toxins (SLT-I and SLT-II), adherence factor of enteropathogenic E. coli (EAF) and invasive factor (INV). Six colonies isolated from 4 food samples hybridized with the probes for LT-II (3 colonies isolated from a milk sample), SLT-I and SLT-II (1 colony isolated from raw bovine meat) or EAF (2 colonies isolated from two raw chicken meat samples).


Assuntos
Sondas de DNA , DNA Bacteriano/análise , Proteínas de Escherichia coli , Escherichia coli/patogenicidade , Microbiologia de Alimentos , Adesinas de Escherichia coli , Animais , Aderência Bacteriana , Proteínas da Membrana Bacteriana Externa/análise , Proteínas da Membrana Bacteriana Externa/genética , Toxinas Bacterianas/análise , Toxinas Bacterianas/genética , Laticínios , Enterotoxinas/análise , Enterotoxinas/genética , Escherichia coli/genética , Escherichia coli/isolamento & purificação , Carne , Produtos da Carne , Leite , Hibridização de Ácido Nucleico , Plasmídeos , Toxina Shiga I , Toxina Shiga II , Virulência
13.
J Infect ; 48(2): 161-7, 2004 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-14720492

RESUMO

The frequency of diarrhoeagenic Escherichia coli (DEC) strains was investigated in 253 children up to 3 years old, with (patient group, PG, 199 children) and without (control group, CG, 54 children) diarrhoea, living in Rio de Janeiro, Brazil. DEC strains were detected in 70 (27.6%) children, including 54 (27.1%) with diarrhoea and 16 (29.6%) without diarrhoea. Enteroaggregative E. coli (EAEC) was the most frequent DEC category, accounting for 14.6% of the isolates in the PG and for 11.1% in the CG. E. coli strains carrying enteropathogenic E. coli (EPEC) virulence markers showed higher incidence in the CG (12.9%) than in the PG (8.0%). E. coli strains belonging to non-classical EPEC groups that carried eae only or eae and bfpA, designated as attaching-effacing E. coli (AEEC) were the most frequent (79.1%). Simultaneous presence of multiple EPEC virulence factors (EAF/eae/bfpA) were only detected among strains isolated from the PG. Enterotoxigenic E. coli (ETEC) strains were isolated from 5.5% of the children in the CG and from 3.5% of those in the PG. Most of the ETEC isolates were LT-probe positive (70%) and none carried both LT-I and ST-I probe sequences. One enteroinvasive E. coli (EIEC) strain was recovered from a child with diarrhoea. No stx-probe positive E. coli strains were detected. Overall, DEC strains were not found to be significantly associated with diarrhoea (p>0.05). However, the higher incidence of EAEC, the most frequent DEC category, among children with diarrhoea, suggests a potential role of EAEC as an important enteric pathogen in the community investigated.


Assuntos
Diarreia/microbiologia , Infecções por Escherichia coli/microbiologia , Escherichia coli/classificação , Testes de Aglutinação , Toxinas Bacterianas/genética , Toxinas Bacterianas/isolamento & purificação , Brasil/epidemiologia , Pré-Escolar , DNA Bacteriano/química , DNA Bacteriano/genética , Diarreia/epidemiologia , Enterotoxinas/genética , Enterotoxinas/isolamento & purificação , Escherichia coli/genética , Escherichia coli/isolamento & purificação , Escherichia coli/patogenicidade , Infecções por Escherichia coli/epidemiologia , Fezes/microbiologia , Feminino , Humanos , Incidência , Lactente , Masculino , Hibridização de Ácido Nucleico , Virulência
14.
Vet Microbiol ; 42(2-3): 105-10, 1994 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-7886925

RESUMO

Forty-six verotoxin-producing Escherichia coli (VTEC) strains isolated from diarrhoeic and healthy calves in Spain were examined for DNA sequences homologous to genes for verotoxins (VT1 and VT2) and enterotoxins (LT-I, LT-II, STaH, STaP and STb). Hybridisation showed that 26 (57%) of VTEC strains carried VT1 genes, 13 (28%) possessed VT2 genes, and 7 (15%) carried both VT1 and VT2 genes. No VTEC strains hybridised with DNA probes for enterotoxins. A correlation was found between the serotype and type of VT produced. Thus, all strains of serotypes O26:K-:H11 (13 strains), O103:K-:H2 (3 strains) and O128:K?:H- (4 strains) hybridised with the VT1 probe only, whereas all strains of serotypes O4:K-:H4 (3 strains) and O113:K-:H21 (4 strains) were positive with the VT2 probe only. By contrast, O81:K?:H28 (2 strains) and O157:K-:H- (2 strains) strains hybridised with both VT1 and VT2 probes. One strain of serotype O157:K-:H7 was VT2 positive.


Assuntos
Toxinas Bacterianas/biossíntese , Toxinas Bacterianas/genética , Bovinos/microbiologia , Escherichia coli/genética , Genes Bacterianos , Animais , Doenças dos Bovinos/microbiologia , Sondas de DNA , Diarreia/microbiologia , Diarreia/veterinária , Reservatórios de Doenças , Enterotoxinas/genética , Escherichia coli/classificação , Escherichia coli/patogenicidade , Humanos , Sorotipagem , Toxina Shiga I , Toxina Shiga II , Espanha
15.
Vet Microbiol ; 83(1): 71-80, 2001 Oct 22.
Artigo em Inglês | MEDLINE | ID: mdl-11524167

RESUMO

Eight Escherichia coli isolates from ostriches with respiratory disease were investigated for the presence of genes encoding the following adhesins: type 1 pili (fim), pili associated with pyelonephritis (pap), S fimbriae (sfa), afimbrial adhesin (afaI), temperature regulated adhesin, curli (crl, csgA) and temperature-sensitive hemagglutinin (tsh). Genes for heat labile (LT) and heat stable (STa and STb) enterotoxins, Shiga toxins (stx1 and stx2), cytotoxic necrotizing factor 1 (cnf), alpha-haemolysin (hly) and aerobactin (aer) production were also investigated. Other characteristics investigated were the presence of hemagglutination activity, growth on an iron-deficient medium, aerobactin production, serum resistance, adherence to chicken tracheal cells, pathogenicity for day-old chicks, and serogroup. Serogrouping showed that four isolates belonged to serogroup O2, two to serogroup O78, one to serogroup O9, and one to serogroup O21. The virulence genes found were: fim in all eight isolates, csgA in seven, aer in six, and pap, crl and tsh in one isolate each. All isolates analyzed were positive for mannose-resistant hemagglutination, adhered in vitro to ciliated tracheal epithelium, grew on iron-deficient medium, and showed serum resistance. Pathogenicity tests on day-old chickens revealed one highly pathogenic isolate, three of low pathogenicity and four isolates with intermediate pathogenicity.


Assuntos
Doenças das Aves/microbiologia , Escherichia coli/patogenicidade , Pulmão/microbiologia , Infecções Respiratórias/veterinária , Struthioniformes , Adesinas de Escherichia coli , Animais , Sequência de Bases , Primers do DNA , Escherichia coli/genética , Fímbrias Bacterianas , Testes de Hemaglutinação/veterinária , Ácidos Hidroxâmicos , Reação em Cadeia da Polimerase/veterinária , Infecções Respiratórias/microbiologia , Sorotipagem/veterinária , Virulência/genética
16.
Vet Microbiol ; 101(4): 269-77, 2004 Aug 06.
Artigo em Inglês | MEDLINE | ID: mdl-15262000

RESUMO

Escherichia coli isolates recovered from 182 fecal specimens from dogs up to five months old from the cities of São Paulo and Campinas, SP, Brazil, were examined by polymerase chain reaction (PCR) for several virulence factors and properties. The eae gene was found in 23 isolates of E. coli from 22 dogs, 19 of 146 (13%) from dogs with diarrhea and 3 of 36 (8.3%) from dogs with no diarrhea. Two different eae+ isolates were recovered from one dog with diarrhea. Isolates from two dogs with diarrhea harbored the bfpA gene, and none of the isolates possessed genes for enterotoxins, the EAF plasmid or Shiga toxins. PCR showed that, among the 23 isolates, eight were positive for beta intimin, six for gamma, two for, one for alpha, one for kappa, and five showed no amplification with any of the nine pairs of specific intimin primers used. PCR also showed that the LEE (locus of enterocyte effacement) was inserted in selC in four isolates, likely in pheU in seven isolates, and in undetermined sites in twelve isolates. Fifteen isolates adhered to HEp-2 cells and were fluorescence actin staining (FAS) positive. The predominant adherence pattern was the localized adherence-like (LAL) pattern. The eae-positive isolates belonged to a wide diversity of serotypes, including O111:H25, O119:H2 and O142:H6, which are serotypes that are common among human EPEC. These results confirmed the presence of EPEC in dogs (DEPEC) with and without diarrhea. The virulence factors found in these strains were similar to those in human EPEC, leading to the possibility that EPEC may move back and forth among human and canine populations.


Assuntos
Aderência Bacteriana , Doenças do Cão/microbiologia , Infecções por Escherichia coli/veterinária , Escherichia coli/patogenicidade , Adesinas Bacterianas/genética , Animais , Sequência de Bases , Brasil , Diarreia/microbiologia , Diarreia/veterinária , Cães , Escherichia coli/genética , Escherichia coli/isolamento & purificação , Infecções por Escherichia coli/microbiologia , Proteínas de Escherichia coli/genética , Fezes/microbiologia , Proteínas de Fímbrias/genética , Genes Bacterianos , Humanos , Reação em Cadeia da Polimerase/veterinária , Sorotipagem/veterinária , Virulência/genética
17.
Braz J Med Biol Res ; 25(7): 667-72, 1992.
Artigo em Inglês | MEDLINE | ID: mdl-1342597

RESUMO

1. The usefulness of plasmid profile analysis to differentiate strains of enteropathogenic Escherichia coli (EPEC) was evaluated by studying 123 strains of the most prevalent serotypes causing infant diarrhea in the city of São Paulo, Brazil, i.e., O111ab:H-, O111ab:H2 and O119:H6. 2. No common profiles were found among strains of distinct serotypes. However, within each serotype, most of the strains were grouped within a few major profiles. More than 68% of the strains of serotypes O111ab:H- and O111ab:H2 were included in 6 and 9 major profiles, respectively. In serotype O119:H6, about 48% of the strains were included in 3 major profiles. 3. This analysis suggests that only a few EPEC clones are causing infant diarrhea in São Paulo and revealed that the distribution of serotypes O111ab:H- and O111ab:H2 during the one-year study was at least partly determined by small outbreaks of the most common profiles. 4. We conclude that plasmid profile analysis is very useful to differentiate strains within specific EPEC serotypes.


Assuntos
Escherichia coli/classificação , Plasmídeos , Brasil/epidemiologia , Diarreia Infantil/epidemiologia , Diarreia Infantil/microbiologia , Escherichia coli/genética , Escherichia coli/isolamento & purificação , Infecções por Escherichia coli/epidemiologia , Infecções por Escherichia coli/microbiologia , Humanos , Lactente , Peso Molecular , Prevalência , Sorotipagem
18.
Arq Gastroenterol ; 35(1): 62-8, 1998.
Artigo em Português | MEDLINE | ID: mdl-9711316

RESUMO

OBJECTS: Persistent diarrhea is a frequent disease in developing countries. In this research, we studied 21 patients that passed away among 189 that were hospitalized with persistent diarrhea at the "Hospital Italo Brasileiro Umberto I Foundation", from January of 1985 until December of 1992. PATIENTS AND METHODS: The patients were distributed into two groups: survival and dead, in accord of the clinical evolution at the end of the internment. The analyzed parameters were: birth weight, sex, age, provenance, diarrhea period before the admission, nutritional status, hydration status, coprologic results, occurrence of food intolerance, internment period and the age of ending breast-feeding. RESULTS: The parameters that showed significantly association with the death were: age, with relative risk = 3 for children with age below 6 months old; provenance, with relative risk = 3.4 for patients who were arrived from other hospitals; third grade dehydration at the admission (relative risk = 2.9); enteropathogenic Escherichia coli (EPEC) isolated in feces (relative risk = 3.3), and use of total parenteral nutrition. The etiologic research was positive in 57.1% of the cases. The enteropathogen more frequently isolated in dead group, was EPEC (42.9%), followed by Shigella (9.5%) and Salmonella (5.9%). From the isolated EPEC (35/189), 26 (74.3%) were belonged to the OIII sorogroup (6/26). From these children, 23.1% died. From the 35 patients with EPEC isolated in feces, 25 were below 6 months old, and from these one, eight died. The relative risk to die for lactents with less than 6 months old and EPEC in feces was equal 3.2. Sepsis was considered the most important cause of death for hospitalized lactents with persistent diarrhea.


Assuntos
Diarreia Infantil/mortalidade , Fatores Etários , Feminino , Humanos , Lactente , Enteropatias/etiologia , Tempo de Internação , Masculino , Fatores de Risco , Desmame
19.
Arq Gastroenterol ; 34(2): 112-20, 1997.
Artigo em Português | MEDLINE | ID: mdl-9496427

RESUMO

Persistent diarrhea very often leads children to malnutrition. It has become the major cause of death resulting from acute diarrhea episodes in developing countries. In order to determine the ultrastructural alterations of the small bowel that occur in the syndrome, 16 infants with severe persistent diarrhea were studied, utilizing light microscopy and the scanning electron microscope. Stool and jejunal fluid samples were collected for culture, rotavirus, ova and parasite search. Enteropathogenic agents were isolated in stools from 11 (68.7%) patients and bacterial proliferation in the small bowel was detected in 11 (68.7%) patients. EPEC strains were the most frequent enteropathogenic agent isolated both from stool and jejunal fluid cultures. The stool cultures revealed the presence of the following enteropathogenic microorganisms: EPEC 0111 in four, EPEC 0119 in one, EAggEC in five, Shigella flexneri in two, and Shigella sonnei in one; mixed infections due to EAggEC associated with EPEC 0111 were seen in two patients. The light microscopic analysis revealed that 56.2% of the patients suffered moderate villous atrophy most frequently associated with effacement of the microvilli, intracytoplasmatic vacuolization, increased number of multivesicular bodied and increased lymphocytic and eosinophylic infiltration in the lamina propria. The scanning electron microscopic analysis revealed in all cases shortening of the villi and enterocyte derangements; very often there was a total lack and/or effacement of the microvilli; in half of the patients there was a mucoid material covering the enterocytes tightly adhered to the apical epithelium surface. The scanning ultrastructural alterations observed in these patients are probably due to an association of factors brought about by the presence of enteropathogenic microorganisms and the resulting food intolerance that is responsible for perpetuation of diarrhea.


Assuntos
Diarreia Infantil/patologia , Intestino Delgado/ultraestrutura , Diarreia Infantil/microbiologia , Duodeno/ultraestrutura , Células Epiteliais/ultraestrutura , Epitélio/ultraestrutura , Feminino , Humanos , Lactente , Masculino , Microscopia Eletrônica de Varredura , Microvilosidades , Estudos Prospectivos
20.
Braz J Med Biol Res ; 44(10): 1018-24, 2011 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-21989977

RESUMO

Although enteropathogenic Escherichia coli (EPEC) are well-recognized diarrheal agents, their ability to translocate and cause extraintestinal alterations is not known. We investigated whether a typical EPEC (tEPEC) and an atypical EPEC (aEPEC) strain translocate and cause microcirculation injury under conditions of intestinal bacterial overgrowth. Bacterial translocation (BT) was induced in female Wistar-EPM rats (200-250 g) by oroduodenal catheterization and inoculation of 10 mL 10(10) colony forming unit (CFU)/mL, with the bacteria being confined between the duodenum and ileum with ligatures. After 2 h, mesenteric lymph nodes (MLN), liver and spleen were cultured for translocated bacteria and BT-related microcirculation changes were monitored in mesenteric and abdominal organs by intravital microscopy and laser Doppler flow, respectively. tEPEC (N = 11) and aEPEC (N = 11) were recovered from MLN (100%), spleen (36.4 and 45.5%), and liver (45.5 and 72.7%) of the animals, respectively. Recovery of the positive control E. coli R-6 (N = 6) was 100% for all compartments. Bacteria were not recovered from extraintestinal sites of controls inoculated with non-pathogenic E. coli strains HB101 (N = 6) and HS (N = 10), or saline. Mesenteric microcirculation injuries were detected with both EPEC strains, but only aEPEC was similar to E. coli R-6 with regard to systemic tissue hypoperfusion. In conclusion, overgrowth of certain aEPEC strains may lead to BT and impairment of the microcirculation in systemic organs.


Assuntos
Translocação Bacteriana/fisiologia , Escherichia coli Enteropatogênica/fisiologia , Infecções por Escherichia coli/microbiologia , Intestinos/microbiologia , Microcirculação , Animais , Criança , Feminino , Humanos , Fígado/microbiologia , Linfonodos/microbiologia , Mesentério/microbiologia , Ratos , Ratos Wistar , Baço/microbiologia
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