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L R Trabulsi

Publications and source records attributed to L R Trabulsi.

At least 73 records · Page 4Linked to original sources

DNA probes for identification of enteroinvasive Escherichia coli.

Eighty-one Escherichia coli strains belonging to all known invasive O serogroups were tested with two distinct invasiveness probes (pMR17 and pSF55). All 54 Sereny test-positive strains and 5 strains that lost Sereny positivity during storage hybridized with both probes. Probe-positive strains carried a 120- to 140-megadalton plasmid, did not produce lysine decarboxylase, and, with the exception of certain serotypes, were nonmotile. Motile strains of serotype O144:H25 were for the first time characterized as invasive by hybridization with the probes.

DNA, Bacterial↗

Plasmids coding for drug resistance and localized adherence to HeLa cells in enteropathogenic Escherichia coli O55:H- and O55:H6.

Plasmids coding for drug resistance and localized adherence (LA) to HeLa cells were found in two enteropathogenic Escherichia coli strains belonging to serotypes O55:H- and O55:H6. Strain 49-81 HSJ (O55:H-) carries two plasmids, one coding for both ampicillin resistance (Apr) and LA (pMS49). Strain 71-82 HSJ (O55:H6) harbors only one plasmid, coding for resistance to sulfadiazine, chloramphenicol, kanamycin, ampicillin, and LA (pMS71). Plasmids pMS49 and pMS71 were transferred to E. coli K-12 711 and from this strain to E. coli K-12 J53. Curing with acridine orange of an Apr LA+ transconjugant showed that both characteristics were lost simultaneously. The plasmids have a molecular weight of approximately 55 X 10(6) and are the first naturally recombinant plasmids coding for adherence and drug resistance described in enteropathogenic E. coli.

Adhesiveness↗

Variation in chemical properties and antigenic determinants among type II heat-labile enterotoxins of Escherichia coli.

Type II heat-labile enterotoxin (LT-II) from Escherichia coli 41 was purified and compared with prototype LT-II encoded by genes from E. coli SA53. Both toxins were oligomeric proteins consisting of polypeptides A (Mr, 28,000) and B (Mr, 11,800). The A polypeptides were cleaved by trypsin into fragments A1 (Mr, 21,000) and A2 (Mr, about 7,000). These two toxins were shown to belong to two different subclasses of LT-II. We propose to designate the prototype toxin LT-IIa and the new variant LT-IIb. The pI of LT-IIb was between 5.2 and 5.6, significantly lower than the pI of 6.8 for LT-IIa, and the behavior of LT-IIb during purification differed significantly from that of LT-IIa. The toxic dose of unnicked LT-IIb in the Y1 adrenal-cell assay was 94 pg, but trypsin-treated, nicked LT-IIb was toxic at about 3 pg. In contrast, the toxic dose of LT-IIa was previously shown to be 0.5 to 1 pg for several preparations that varied from unnicked to partially nicked, and treatment with trypsin was not required for full toxicity. The titer of LT-II antiserum in neutralization tests was 100-fold greater against LT-IIa than against LT-IIb. In immunodiffusion tests, LT-IIa and LT-IIb gave a reaction of partial identity. In a radioimmunobinding assay, the titer of LT-IIa antiserum against homologous LT-IIa was approximately 10-fold greater than against LT-IIb. The cholera-E. coli family of heat-labile enterotoxins has been divided into serogroup I, which includes cholera toxin and the antigenic variants of E. coli heat-labile toxin designated LTh-I and LTp-I, and serogroup II, which includes LT-IIa and LT-IIb. The type I and type II toxins do not cross-react in neutralization or immunodiffusion tests. By using very sensitive radioimmunobinding assays, it was possible to demonstrate common antigenic determinants between the type I and type II toxins. However, the titers of antibodies in hyperimmune sera that recognized these common determinants were very low.

Antigen-Antibody Complex↗

Production of type II heat-labile enterotoxin by Escherichia coli isolated from food and human feces.

Escherichia coli strains isolated in Sao Paulo, Brazil, from feces of patients with diarrhea and from food samples produced toxin(s) that was shown to be related both immunologically and genetically to the recently characterized type II heat-labile enterotoxin of E. coli. The new isolates of type II heat-labile enterotoxin-producing E. coli belonged to five different serotypes and did not represent a single clone.

Bacterial Toxins↗

Enterotoxin production, presence of colonization factor antigen I, and adherence to HeLa cells by Escherichia coli O128 strains belonging to different O subgroups.

Strains of three subgroups of Escherichia coli O128 were studied. Enterotoxin production was observed in 30 (91%) O128ac strains, whereas strains of subgroups O128ab and O128ad were not toxigenic. CFA/I was only found in two serotypes of subgroup O128ac, all of them producing heat-stable enterotoxin except for one which produced both toxins. None of the strains studied produced CFA/II. In a binding test with HeLa cells, localized adherence was found only in strains of subgroup O128ab; diffuse adherence occurred in strains of subgroup O128ac. As flagellar antigens were specific in subgroups ab and ac and toxin production was observed only in subgroup ac, the present results suggest that subgroup and serotype are useful markers for O128 strains that are enterotoxigenic or enteropathogenic.

Antigens, Bacterial↗

Plasmid-mediated factors conferring diffuse and localized adherence of enteropathogenic Escherichia coli.

Histopathological evidence suggests that the adherence of enteropathogenic Escherichia coli (EPEC) to the mucosa of the small bowel is an important step in pathogenesis. Several reports have shown that many EPEC isolates adhere to HEp-2 and HeLa cells in tissue cultures. In the HeLa cell assay, there are at least two distinct patterns of adherence: localized adherence, which is characterized by the formation of bacterial microcolonies, and diffuse adherence, in which bacteria cover the cell uniformly. We have found that these two patterns can be demonstrated in HEp-2 cells as well as in HeLa cells and that the results of the two assays are closely correlated. Using a DNA probe which is sensitive and specific for localized adherence to HEp-2 cells, we provide evidence that localized adherence and diffuse adherence by EPEC are due to at least two genetically distinct adhesions which confer phenotypic differences in both the morphology of HEp-2 cell adherence and in surface hydrophobicity. The two factors are each encoded on plasmids which vary in size from 55 to 70 megadaltons; one strain exhibiting localized adherence carried these genes on the chromosome.

Adhesins, Escherichia coli↗

Detection of genes for heat-stable enterotoxin I in Escherichia coli strains isolated in Brazil.

Heat-stable enterotoxin I (STI) can be assayed in intestinal loops of pigs and rabbits and in the gut of infant mice. To produce a simpler and more discriminating assay procedure, we used three gene probes corresponding to three forms of STI called STIa, STIb, and STIc. We tested 159 Brazilian isolates, of which 40 were positive in the infant mouse assay. The STIb and STIc probes are similar (93% DNA homology) and are both different from the STIa probe (70% DNA homology). Of 33 strains that were still active for STI 3 years after their isolation, 25 reacted with both the STIb and STIc probes, 4 reacted with the STIc probe only, and 7 reacted strongly with the STIa probe and weakly or not at all with the other probes. Two strains reacted with all three probes. Further analysis showed that each of these two strains contains a small plasmid that reacts with the STIa probe and a large plasmid that reacts with the STIc probe in one strain and weakly with both the STIa and STIc probes in the other strain. It was also shown that the STIa probe reacts with the cloning vehicle pACYC184 used for the cloning of STIc. We conclude that the gene probes used can identify most STI-producing strains and that in cases of positive responses with several probes careful scrutiny is necessary for analysis.

Bacterial Toxins↗

Correlation between adherence to HeLa cells and serogroups, serotypes, and bioserotypes of Escherichia coli.

Four hundred fifty Escherichia coli strains of 45 O serogroups and subgroups and 112 serotypes were studied to determine their patterns of adherence to HeLa cells. Adherence was exhibited by strains of 17 O serogroups and subgroups, but within these groups more than one adherence pattern was frequently observed. However, within each serotype, the adherence pattern was highly consistent. Localized adherence (LA) was observed much more frequently in serotypes that we considered to be enteropathogenic E. coli serotypes (93%) than in other serotypes (14%), whereas diffuse adherence (DA) occurred predominantly among nonenteropathogenic E. coli strains. Determination of biochemical characteristics showed that within O serogroups, nonmotile strains tended to have the same behavior as motile strains with the LA adherence pattern, suggesting that they were derived from these motile strains. LA and non-LA strains of the same serotype differed biochemically. LA appears to be a property of most E. coli commonly considered to be enteropathogenic and should assist attempts to determine which E. coli are enteropathogenic and to elucidate their pathogenic mechanisms.

Adhesiveness↗

Evaluation of antisera used for detecting enterotoxigenic Escherichia coli in Sao Paulo.

The usefulness of antisera in detecting enterotoxigenic Escherichia coli (ETEC) strains in Sao Paulo was evaluated. Polyvalent antisera detected 49% of ETEC isolates and were more effective in identifying E. coli that produced heat-labile and heat-stable enterotoxins and in strains that produced only heat-stable enterotoxin. ETEC strains not detected by the antisera belonged to different serogroups not isolated in Sao Paulo before; 34% of these strains had undetermined O antigens, and most of them produced only heat-labile toxin. A variation of serogroups over time was especially observed among strains that produced heat-stable toxin. The importance of H-antigen determinations in the effectiveness of ETEC diagnosis by serological methods became evident, as non-ETEC strains were also detected by polyvalent antisera, but their serotypes were different from those of ETEC strains. Although antisera can be used to identify O:H types of ETEC strains with accuracy, serotyping cannot be recommended for routine diagnosis. However, such a procedure may be useful for studying outbreaks of ETEC diarrhea if the involved serotypes are already known.

Animals↗

Experimental guinea pig ocular infection by Salmonella typhimurium.

The clinical, microbiologic, and cytologic features of the guinea pig model of keratoconjunctivitis with enterobacteria, Salmonella typhimurium were elucidated. Guinea pig eyes were instilled with S. typhimurium and the eyes were studied by biomicroscopy, culture, cytology, pathology, and electron microscopy. All animals developed moderate to severe conjunctivitis that was present in 18% of the animals on day 1. It became more intense, appearing in all of the eyes on day 10 and disappeared before day 30. The cultures for S. typhimurium were almost all positive on days 1 and 2, declined steadily to 10% on day 10, and were negative after that. A coarse, epithelial punctate keratitis was present in more than 90% of the infected eyes at some time during the experiment. The keratitis had a biphasic clinical course. The first peak correlated with the maximum culture results, but during the second peak only 10% of the cultures were positive. Electron microscopy of the cornea showed the S. typhimurium at the epithelial surface within surface epithelial cells during the early phases of infection. The later phase keratitis, with negative culture results, resembles the keratitis of Reiter's syndrome.

Animals↗

A controlled study of endemic sporadic diarrhoea among adult residents of southern Brazil.

The aetiology of sporadic summer diarrhoea and enterotoxin (LT) antibody titres was studied among 43 adult patients in southern Brazil who had an acute, untreated diarrhoeal illness and 43 age- and sex-matched controls from the same area. A potential pathogen was identified in 33 of 34 patients and in 17 of 43 controls (p less than 0.01). 10 Shigella, one invasive Escherichia coli, one Salmonella and one Entamoeba histolytica were identified in 17 patients with inflammatory diarrhoea (faecal polymorphonuclear neutrophil leucocytes (PMN) present). In the other four only enterotoxigenic strains of E. coli were identified. Among 26 patients with non-inflammatory (no faecal PMN) diarrhoea, heat-stable (ST) toxin-producing coliforms were the most common pathogens isolated (27%). Heat-labile (LT) toxin-producing coliforms or serological rises to LT were seen in five patients, Strongyloides larvae in five, Giardia lamblia trophozoites in four and an LT-producing Salmonella in one. No pathogen was found in 10 patients with non-inflammatory diarrhoea. Among 43 controls, 11 LT coliforms, two Shigella and two Strongyloides larvae were found. 40 of 41 controls had detectable serum anti-LT antibody. A single ST-producing E. coli was found in one other control. No pathogen was identified in 26 controls. No rotaviruses or Norwalk-like viral agents were seen by direct or immune electron microscopy of stool filtrates. While LT-producing E. coli are frequently associated with endemic summer childhood diarrhoea in southern Brazil and are recognized causes of travellers' diarrhoea, this study shows that they are often carried asymptomatically by adults living in this region.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

The significance of hospitals as reservoirs for endemic multiresistant Salmonella typhimurium causing infection in urban Brazilian children.

To identify possible sources of multiply drug-resistant Salmonella typhimurium among children in São Paulo, Brazil, we reviewed records of 470 children who had visited eight outpatient clinics from March 1981 to May 1982 and of 28 children who had been admitted to one referral hospital between June and November of 1982, and we examined plasmid profiles of the Salmonella isolates by agarose-gel electrophoresis. S. typhimurium was identified in 37 of these children. Case-control studies showed that children with S. typhimurium infections were more likely to have been hospitalized before onset of diarrhea than were either age-matched children without diarrhea (P = .031) or age-matched children with nonbacterial diarrhea (P = .035). Four distinct plasmid profiles, each of which was temporally clustered, were identified in 20 (67%) of 30 S. typhimurium strains isolated from previously hospitalized children and in two (28%) of seven strains from children not previously hospitalized. Each of three of these four profiles was associated with a different hospital, results suggesting that multiresistant-S. typhimurium infections in São Paulo are often nosocomially acquired.

Anti-Bacterial Agents↗

Distinctive patterns of adherence of enteropathogenic Escherichia coli to HeLa cells.

We showed that Escherichia coli strains attach to HeLa cells in two different patterns. In one, the bacteria cover the whole surface of the cell (diffuse adherence), and in the other, attachment is limited to one or a few sites of the cell surface (localized adherence). Among the enteropathogenic strains, serogroups O55, O86, O11ab, O119, O125, O128ab, and O142 usually showed localized adherence when tested in the presence of D-mannose. Localized adherence was not shown either by E. coli strains isolated from urine or by enteroinvasive and enterotoxigenic E. coli strains. Some of these strains showed diffuse adherence. Some strains of serogroups O55, O111, and O119 showed both localized and diffuse adherence in the same preparation. Mannose-resistant adherence was not related to colonization factor antigens.

Adhesiveness↗

Enteropathogenic Escherichia coli serotypes and endemic diarrhea in infants.

Enteropathogenic Escherichia coli serotypes were searched for in feces of 550 children with endemic diarrhea and in 129 controls, in São Paulo, in 1978 and 1979; serotypes O111ab:H(-), O111ab:H(2), and O119:H6 were significantly associated with diarrhea in children 0 to 5 months old and were the most frequent agents of diarrhea in this age group as compared with enterotoxigenic and enteroinvasive E. coli, Salmonella sp., Shigella sp., and Yersinia enterocolitica. It is concluded that various enteropathogenic E. coli serotypes may be agents of endemic infantile diarrhea.

Diarrhea↗

Plasmid coding for drug resistance and production of heat-labile and heat-stable toxins harbored by an Escherichia coli strain of human origin.

In a study of enterotoxigenic strains of Escherichia coli isolated from children with diarrhea in São Paulo, Brazil, a new enterotoxin and antibiotic resistance plasmid that carries heat-labile toxin, heat-stable toxin, and drug resistance genes was found. This is the first such plasmid to be found in a human strain of E. coli. The plasmid is nonconjugative, has a molecular weight of at least 54 x 10(6), and is mobilized by the R plasmid present in the host strain.

Anti-Bacterial Agents↗