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L C Campos

Publications and source records attributed to L C Campos.

13 recordsLinked to original sources

Genetic diversity and antibiotic resistance of clinical and environmental Vibrio Cholerae suggests that many serogroups are reservoirs of resistance.

Vibrio cholerae is an important human pathogen and the cause of cholera. Since genetic variation and antibiotic resistance of strains have implications for effective treatment of the disease, we examined the genetic diversity and antibiotic resistance profile in 92 clinical strains (serogroup O1) and 56 environmental strains (O1 antigen, 42 strains; non-O1 antigen, 14 strains) isolated in Brazil between 1991 and 1999. Clinical and environmental O1 strains showed greater drug resistance compared to environmental non-O1 strains. Nearly all clinical O1 strains were resistant to one or more antibiotics while half of the environmental O1 and non-O1 strains were resistant to one or more antibiotics. No plasmids or class 1 integrons were detected in the strains by PCR analysis. Multilocus enzyme electrophoresis analysis (MLEE) suggests most of the O1 strains belong to a single (South American) clone that is related but different to seventh-pandemic strains isolated from other parts of the world. Our results show that there is a close genetic relationship between clinical and environmental O1 strains and that many serogroups and the environment can be a reservoir for antibiotic resistance.

Anti-Infective Agents↗

Chemical composition and microbicidal activity of extracts from Brazilian and Bulgarian propolis.

AIMS: The chemical composition of ethanol extracts from a Brazilian (Et-Bra) and a Bulgarian (Et-Blg) propolis, and their activity against the protozoan Trypanosoma cruzi, several fungi and bacteria species were determined. METHODS AND RESULTS: The chemical composition was determined by high temperature high resolution gas chromatography coupled to mass spectrometry. Microbiological activity was assayed in vitro against T. cruzi, Candida albicans, Sporothrix schenckii, Paracoccidioides brasiliensis, Neisseria meningitidis, Streptococcus pneumoniae and Staphylococcus aureus. CONCLUSIONS: Et-Bra and Et-Blg, although with totally distinct compositions, were active against T. cruzi and the three species of fungi. Et-Blg was more effective than Et-Bra against bacteria, particularly N. meningitidis and Strep. pneumoniae. SIGNIFICANCE AND IMPACT OF THE STUDY: Although with different classes of components, both propolis extracts showed microbicidal activity. For the bactericidal activity it was possible to establish a positive correlation with the high content of flavonoids of the Bulgarian extract.

Animals↗

Biomass development in slow sand filters.

Microbial biomass development in the sand and schmutzdecke layer was determined in two full-scale slow sand filters, operated with and without a light excluding cover. A standard chloroform fumigation-extraction technique was adapted to routinely measure microbial biomass concentrations in the sand beds. Sand was sampled to a depth of 10 cm and schmutzdecke was also collected at the same random positions on the uncovered filter. Interstitial microbial biomass in the uncovered sand bed increased with time and decreased with sampling depth. There was a small accumulation of sand biomass with time in the covered filter, but no relationship was apparent between biomass concentration and depth in this filter. Schmutzdecke did not develop on the covered filter and was spatially highly variable in the uncovered condition compared to the consistent patterns observed in interstitial biomass production. It is speculated that microbial biomass in the sand of uncovered filters is largely related to carbon inputs from photosynthetic activity in the schmutzdecke and involves mechanisms that spatially distribute carbon substrate from the schmutzdecke to the sand. However, total organic carbon and dissolved organic carbon removals were similar in both filters suggesting that relatively small biomass populations in covered filters are sufficient to remove residual labile carbon during advanced water treatment and little further advantage to water purification and organic carbon removal is gained by the increased production of biomass in uncovered slow sand filter beds.

Biomass↗

Diffusely adhering Escherichia coli (DAEC) strains of fecal origin rarely express F1845 adhesin.

A total of 398 diffusely adhering Escherichia coli (DAEC) strains of fecal origin were analyzed for the presence of sequences homologous to the structural subunit gene (daaE) of the F1845 fimbria. For that purpose, a DNA fragment homologous to daaE, obtained by PCR, was used as a probe in colony hybridization assays. Only two strains carried daaE and expressed F1845, suggesting that this fimbria is rare among DAEC strains.

Adhesins, Escherichia coli↗

Virulence properties of Escherichia coli O111:H12 strains.

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.

Animals↗

Virulence properties and clonal structures of strains of Escherichia coli O119 serotypes.

A total of 110 Escherichia coli strains of serogroup O119 were examined for the presence of virulence properties characteristic of enteropathogenic E. coli (EPEC). Three virulence patterns were distinguished based on the detection of a chromosomal gene mediating intimate attachment (eaeA) and plasmid DNA involved in localized adherence (EAF and bfpA). The first pattern, represented by strains which hybridized with three gene probes, was the most common (68%) and, with a single exception, included only O119:H6 strains. Of these strains, 90% showed a typical localized adherence (LA) pattern in HEp-2 cells and 96% were positive for intimate attachment in a fluorescent-actin staining test with a 3-h incubation period. The second pattern was represented by strains which hybridized with the eaeA gene only. Most (89.5%) of these strains showed the LA phenotype but only after 6 h of incubation (LA-like phenotype). The third pattern consisted of strains which were positive for eaeA and bfpA but did not hybridize with the EAF probe. Most (80%) of these strains exhibited the LA-like phenotype. Analysis of several eaeA+ bfpA+ strains for the expression of the pilin subunit (BfpA) of the bundle-forming pili demonstrated that all LA strains expressed BfpA whereas the LA-like strains did not. The study of the clonal relationships, carried out by multilocus enzyme electrophoresis in 79 representative strains, defined 11 distinct electrophoretic types (ETs). ET1 included 66% of the strains, most of which displayed the eaeA+ bfpA+ EAF+ pattern and were serotyped as O119:H6 or O119:H-. The remaining 10 ETs were each represented by no more than five strains and, with the exception of ET8, included strains of a single serotype. The genetic relatedness of the ETs revealed two main clusters, with most strains in cluster A having the eaeA+ bfpA+ EAF+ combination and a O119:H6 serotype. Cluster B was represented by atypical EPEC strains with only the eaeA+ and the eaeA+ bfpA+ virulence pattern.

Bacterial Adhesion↗

Clonal structure and virulence factors in strains of Escherichia coli of the classic serogroup O55.

Virulence properties and genetic variation as determined by multilocus enzyme electrophoresis were studied in 70 strains of Escherichia coli 055, a common serogroup of enteropathogenic E. coli (EPEC), a major cause of infantile diarrhea in developing countries. Nearly 40% of the strains were originally isolated in Brazil and represented serotypes 055:H6, 055:H7, and 055:H51 and nonmotile (055:H-) strains. The analysis of electrophoretic variants of 20 enzymes defined seven distinct electrophoretic types (ETs). ET 1 was represented by 41% of the strains, including strains which usually hybridized with DNA probes for the intimin gene (eaeA), the EPEC adherence plasmid (EAF), and the gene for the pilin subunit of the bundle-forming pilus (bfpA). The ET 1 strains were also typically serotype 055:H6, displayed localized adherence (LA) in tissue culture assays, and were positive in the fluorescent-actin staining test for intimate cell adherence. These same characteristics were observed in the closely related ETs 2 to 4, which clustered in the same branch as ET 1. No known virulence marker could be identified in ET 6. ET 5 included 23 strains, all of which carried the eaeA gene but otherwise displayed a striking array of distinct virulence traits. This ET was represented by 055:H7 strains with phenotypes as diverse as the simultaneous expression of LA and diffuse adherence and the ability to form a newly described adherence pattern, called LA-like adherence. The results suggest that ET 5 marks a special pathogenic clone with a propensity to acquire virulence factors which may facilitate the emergence of new pathogenic strains.

Brazil↗

Diffuse adherence, ST-I enterotoxin and CFA/IV colonization factor are encoded by the same plasmid in the Escherichia coli O29:H21 strain.

Escherichia coli O29:H21 is a human enterotoxigenic serotype that produces heat-stable (ST-I) enterotoxin, adheres diffusely to HeLa cells, and presents colonization factor antigen IV (CFA/IV) composed of CS5CS6 surface antigens. In one strain studied the genes for diffuse adherence and CFA/IV (CS5CS6) production were found to be present in the same plasmid encoding ST-I. The virulence plasmid (Ent) presented two unrelated basic replicons homologous to repFIC and repW. Gene(s) encoding diffuse adherence did not share homology with the probe for F1845 fimbrial adhesin which is responsible for this phenotype in other E. coli strains. Ent plasmids containing genes for diffuse adherence have not been described previously.

Acridine Orange↗

Escherichia coli serogroup O111 includes several clones of diarrheagenic strains with different virulence properties.

Genetic variation among isolates of Escherichia coli O111 obtained mostly from patients with diarrhea in Brazil was assessed by multilocus enzyme electrophoresis to characterize chromosomal genotypes and by gene probes and adherence assays to characterize virulence properties. Among the 152 isolates, we resolved 16 distinct electrophoretic types (ETs), which differed on average at 40% of the enzyme loci. We identified four major bacterial O111 clones of different disease classes: ET 12, which includes the bulk of the enteropathogenic E. coli strains, typically showing localized adherence and intimate attachment in tissue culture assays; ET 1, which includes strains with a different set of virulence markers; ET 9, which includes strains that show intimate attachment but lack localized adherence and Shiga-like toxin genes; and ET 8, which includes strains that are Shiga-like toxin producers and have the corresponding traits of enterohemorrhagic E. coli. Enteroaggregative strains constituted ET 10 and also occurred in ET 1. Isolates of the major clones were found in South and North America and matched in ET and virulence factors to previously described diarrheagenic clones that are widely disseminated in the human population. Because the major clones are genetically distantly related and exhibit different combinations of virulence factors, we hypothesize that they have distinct mechanisms of pathogenesis. The results indicate that genetic divergence of bacteria with the O111 antigen, as measured by allelic variation in enzyme loci, is accompanied by divergence in virulence properties of clones so that identification and classification of pathogenic E. coli strains cannot be based solely on serotyping or a single virulence factor.

Bacterial Adhesion↗

In vitro susceptibility of Salmonella to ciprofloxacin and pefloxacin compared with three other antibiotics.

The in vitro susceptibilities of 703 clinical isolates of Salmonella to ciprofloxacin (CIP) and pefloxacin (PEF) were compared with those to trimethoprim-sulfamethoxazole (TMP-SMZ), chloramphenicol (CO) and ampicillin (AP). All isolates were susceptible to CIP, while PEF inhibited 90.7% of the strains. In contrast, resistance rates of 40, 29.2 and 27% were detected for AP, TMP-SMZ and CO, respectively. PEF resistance was detected in S. panama (1), S. typhi (3) and S. typhimurium (17), the latter representing the most frequently serovar isolated in our country. None of the S. typhi isolates was resistant to CO. Combined resistance was most frequently found among S. typhimurium isolates, with the patterns PEF-TMP-SMZ-AP (10) and PEF-TMP-SMZ-CO-AP (5) predominating.

Ampicillin↗

Antimicrobial resistance among Salmonella serovars isolated from different sources in Brazil during 1978-1983.

A total of 748 Salmonella strains (97 serovars) isolated from human (291), animal (119), environmental (141), food (102) and animal feed (95) sources were examined for resistance to 9 antimicrobial agents. Most of the human isolates were from hospitalized patients (282). An overall resistance rate of 98.8% was determined with 100% for human and environmental isolates. Resistance to sulfadiazine (87.7%) was most common, followed by streptomycin (61.2%), ampicillin (39%) and trimethoprim-sulphamethoxazole (37.9%). Fifty one different resistance patterns were identified with Su (164 strains), Su-Sm (122) and Su-Sm-Tc-Cm-Km-Ap-Nx-Gm-Tm (95) predominating, the latter occurring only in human isolates. Multiple resistance was most frequently found among human isolates, particularly in S. derby and S. typhimurium strains. The relationship between antibiotic resistance, serovar and source of isolation of the Salmonella strains is discussed.

Animals↗

Colicinogeny in Salmonella serovars isolated in Brazil.

A study of colicinogeny was made in 748 strains of Salmonella (97 serovars) isolated from different sources: human (291), animal (119), environmental (141), food (102) and animal feed (95). Colicin production was detected in 64 strains (8.6%), particularly isolated from foods (30.4%). Col E1 (53) and Ia (44) were the most frequently observed, especially in S. agona for environment and food sources. Col V production was identified in 5 strains of S. typhimurium within 8 producer cultures isolated from humans. Its relationship with the sources and serovars of Salmonella are discussed.

Animals↗