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Biomedical subjects

L M Teixeira

Publications and source records attributed to L M Teixeira.

At least 37 records · Page 2Linked to original sources

Leuconostoc pseudomesenteroides as a cause of nosocomial urinary tract infections.

The phenotypic and genotypic characterization of five clinical isolates of Leuconostoc pseudomesenteroides associated with nosocomially acquired urinary tract infections is described. All the strains were susceptible to chloramphenicol, clindamycin, erythromycin, gentamicin, and tetracycline; all were resistant to nalidixic acid, norfloxacin, and vancomycin; and all were intermediately affected by ampicillin and penicillin. Analysis of chromosomal DNA by pulsed-field gel electrophoresis after treatment with SmaI indicated a clonal relationship of the isolates. The results provide evidence for the possibility of nosocomial transmission of this unusual opportunistic, vancomycin-resistant pathogen.

Adolescent↗

Genetic relationships among the different phenotypes of Streptococcus dysgalactiae strains.

The species Streptococcus dysgalactiae was proposed to accommodate a heterogeneous group of streptococci associated with infections in animals and human beings. This taxon is now considered to include animal isolates of alpha-haemolytic group C streptococci, previously called S. dysgalactiae; animal and human isolates of beta-haemolytic group C streptococci, previously called 'S. equisimilis'; beta-haemolytic group L strains associated with infections in animals and, rarely, in humans; and beta-haemolytic group G strains isolated from humans. DNA-DNA reassociation experiments (hydroxyapatite method) and multilocus enzyme electrophoresis (MEE) were performed on reference strains and clinical isolates to determine the genetic relationships among these different phenotypic categories. DNA-DNA hybridization tests showed that they were related at the species level, despite the phenotypic and host heterogeneity. Both genotypic and phenotypic characterization indicated that S. dysgalactiae could be separated into two major sub-groups. The first sub-group contained alpha-haemolytic strains that showed levels of DNA relatedness with the type strain of S. dysgalactiae ranging from 84 to 90% and from 82 to 88% under optimal (55 degrees C) and stringent (70 degrees C) conditions, respectively. The second sub-group contained beta-haemolytic strains showing levels of relatedness ranging from 71 to 79% (55 degrees C) and from 62 to 73% (70 degrees C). Percentage divergence varied from 0.5 to 1.0% (alpha-haemolytic group) and from 2.0 to 3.5% (beta-haemolytic group). A dendrogram based on phenotypic similarity between the enzyme bands produced by MEE showed a Jaccard similarity coefficient of 0.45 between the subclusters formed by the two sub-groups. The results of phenotypic and genotypic characterization were consistent with a published proposal to divide S. dysgalactiae into two subspecies, S. dysgalactiae subsp. dysgalactiae and S. dysgalactiae subsp. equisimilis, with a few modifications.

Animals↗

Use of tests for acidification of methyl-alpha-D-glucopyranoside and susceptibility to efrotomycin for differentiation of strains of Enterococcus and some related genera.

A total of 107 Enterococcus strains, 10 Vagococcus fluvialis strains, and 8 Lactococcus garvieae strains were tested for acidification of methyl-alpha-D-glucopyranoside (MGP) and susceptibility to 100-microg efrotomycin (EFRO) disks. All 26 strains of Enterococcus casseliflavus, including 3 nonmotile and 2 nonpigmented strains, acidified MGP and were resistant to EFRO. All 22 strains of Enterococcus gallinarum, including 5 nonmotile strains, also acidified MGP and were resistant to EFRO. None of the 26 strains of Enterococcus faecium acidified MGP, and all were susceptible to EFRO. Although all 12 Enterococcus faecalis strains were also negative in the MGP test, they were resistant to EFRO. Other enterococcal strains gave variable results. All 10 strains of V. fluvialis and all 8 strains of L. garvieae gave positive and negative results, respectively, in the MGP test and were, respectively, resistant and susceptible to EFRO. These results indicate that tests of the production of acid from MGP and susceptibility to EFRO can be used as adjunct tests in the identification of typical and atypical strains of enterococci in the clinical microbiology laboratory.

Anti-Bacterial Agents↗

Detection and differentiation of vanC-1, vanC-2, and vanC-3 glycopeptide resistance genes in enterococci.

The VanC phenotype, as found in Enterococcus gallinarum, E. casseliflavus, and E. flavescens, is characterized by intrinsic low-level resistance to vancomycin. The nucleotide sequences of the vanC-1 gene in E. gallinarum, the vanC-2 gene in E. casseliflavus, and the vanC-3 gene in E. flavescens have been reported, although there is some disagreement as to whether E. flavescens is a legitimate enterococcal species. Previous attempts to differentiate the vanC-2 and vanC-3 genes by PCR analysis have been unsuccessful. The purpose of the present study was to detect and differentiate the three vanC determinants and examine the distribution of these genes in a collection of both typical and atypical enterococci. The 796-bp vanC-1 PCR product was amplified only from E. gallinarum isolates. As expected, due to the extensive homology in the vanC-2 and vanC-3 gene sequences, all of the E. casseliflavus and E. casseliflavus/flavescens isolates produced the 484-bp vanC-2 PCR product, although the E. gallinarum isolates were negative. Only the E. casseliflavus/flavescens isolates produced the 224-bp vanC-3 product. Using the three sets of primers, we were able to detect and distinguish the vanC-1, vanC-2, and vanC-3 genes from both typical and atypical enterococci strains. Antimicrobial susceptibility tests and analysis of genomic DNA by pulsed-field gel electrophoresis were also performed, but the results indicated that they were not able to distinguish among strains possessing the three vanC genotypes.

Anti-Bacterial Agents↗

Aerococcus urinae: intraspecies genetic and phenotypic relatedness.

A number of Aerococcus-like organisms were recently recognized as human pathogens. Five Aerococcus-like strains were proposed as members of the new species Aerococcus urinae (with type strain E2 [= NCTC 12142]) on the basis of the results of a 16S rRNA sequence analysis. The intraspecies phenotypic and genetic relatedness of 22 selected A. urinae strains was investigated, and a hitherto unrecognized esculin hydrolysis-positive biotype was identified. A total of 14 of the 15 more common esculin-negative strains exhibited very high DNA relatedness as determined by the hydroxyapatite method (the levels of relatedness were greater than 90% in 55 and 70 degrees C reactions, with 1.5% or less divergence in related sequences). The DNA relatedness among the six esculin-positive strains was more heterogeneous, and two DNA hybridization subgroups were formed. Our results are compatible with the hypothesis that both biotypes are members of the single species A. urinae, which contains two or more genetic subspecies. The putative subspecies have not been formally proposed since they cannot be definitively differentiated. The inclusion of A. urinae in the genus Aerococcus is supported by the results of 16S rRNA sequencing. The rRNA sequence data also is compatible with placing both biotypes in a single species.

Bacteriological Techniques↗

Phenotypic and genotypic characterization of Vagococcus fluvialis, including strains isolated from human sources.

This study presents phenotypic and genotypic data for seven isolates of Vagococcus fluvialis, including four strains recovered from human clinical sources, one strain isolated from an environmental source, and two strains isolated from pigs. On the basis of phenotypic characteristics, most isolates were initially classified as "unidentified enterococci," because they resembled atypical arginine-negative enterococcal species. All seven strains as well as the type strain of V. fluvialis reacted with the AccuProbe Enterococcus genetic probe. The seven isolates had virtually indistinguishable whole-cell protein profiles that were similar to that of the V. fluvialis type strain and distinct from those of Enterococcus and Lactococcus species. DNA-DNA reassociation experiments confirmed that the strains were V. fluvialis. They were 71% or more related to the V. fluvialis type strain under optimum and stringent conditions, with 2.5% or less divergence within related sequences. All strains were susceptible to ampicillin, cefotaxime, trimethoprim-sulfamethoxazole, and vancomycin and were resistant to clindamycin, lomefloxacin, and ofloxacin. Strain-to-strain variation was observed in relation to susceptibilities to 18 other antimicrobial agents. Chromosomal DNA was analyzed by pulsed-field gel electrophoresis (PFGE) after digestion with SmaI. Distinctive PFGE patterns were generated, suggesting the nonclonal nature of V. fluvialis strains. Although the number of strains was small, this report provides molecular characterization of V. fluvialis and the first evidence of a possible connection of this species with human infections.

Animals↗

Phenotypic and genotypic characterization of atypical Lactococcus garvieae strains isolated from water buffalos with subclinical mastitis and confirmation of L. garvieae as a senior subjective synonym of Enterococcus seriolicida.

During a survey of bacterial agents that cause subclinical mastitis in water buffalos, we isolated several strains of gram-positive cocci that appeared to be enterococci except that they grew very slowly at 45 degrees C and grew slowly in broth containing 6.5% NaCl. On the basis of the results of conventional physiologic tests, these strains were identified as Enterococcus durans. However, none of the strains reacted with the AccuProbe Enterococcus genetic probe. The whole-cell protein profiles of these organisms were compared with the profiles of Enterococcus and Lactococcus reference strains. apart from minor quantitative differences, the mastitis isolates had indistinguishable protein profiles that were similar to the profiles of the Lactococcus garvieae and Enterococcus seriolicida type strains. The results of DNA relatedness studies performed by using the hydroxyapatite method at 55 and 70 degrees C indicated that all of the mastitis isolates were related to the type strain of L. garvieae at the species level, despite the fact that they exhibited several uncommon phenotypic characteristics (growth at 45 degrees C, growth in broth containing 6.5% NaCl, and failure to produce acid from mannitol and sucrose). The high levels of DNA relatedness between strains of L. garvieae is a senior synonym of E. seriolicida, L. garvieae should be retained as the species name and strain ATCC 43921 should remain the type strain of this species.

Animals↗

Hemagglutination properties of Enterococcus.

In total, 86 enterococcal strains including representatives of most of the described species were tested for the ability to agglutinate human, sheep, and rabbit erythrocytes. Five strains did not react with any of the erythrocytes tested, and 81 (94.2%) strains agglutinated only rabbit erythrocytes. The hemagglutination titers ranged from 2 to 64. Loss of the hemagglutination activity was observed when rabbit erythrocytes were treated with trypsin or neuraminidase. Trypsin treatment of the bacterial suspensions also caused loss of the agglutination ability. On the other hand, heat treatment of bacterial suspensions increased the efficiency of the interactions, and higher titers were obtained. Assays for inhibition of hemagglutination were performed with alpha-D-fucose, alpha-D-galactose, beta-D-galactose, D-glucose, N-acetyl-galactosamine, N-acetyl-glucosamine, N-acetylneuraminic acid, N-acetylneuraminic acid-lactose, and fetuin. Only fetuin was able to inhibit the hemagglutination reactions. The results showed that hemagglutination properties are common to the different enterococcal species tested. They also suggest that enterococci possess hemagglutinins of proteic and non-proteic nature that are involved in the attachment to sialic acid-containing receptors on the surface of rabbit erythrocytes.

Adhesins, Bacterial↗

The electrokinetic surface of five enteropathogenic Escherichia coli serogroups.

The surface ionogenicity of five enteropathogenic Escherichia coli serogroups (O111:H2, O111:H12, O125:H9, O119:H6, and O26:H11) was investigated by electrokinetical approaches. All of the studied surfaces are negatively charged with their mean values of zeta potential (ZP) varying from -9.0 (O26:H11) to -11.9 mV (O111:H2). The populational behavior of the all bacteria are similar since very high ZP values varying from -26 to -30 mV were obtained in experiments carried out with the slip plane calculated at 6.83 nm from the cell surface. All the surfaces are extremely acidic, because the isoelectrophoretic points are localized at pH values below 3.0. Treatment of the microorganisms with neuraminidase did not alter their surface anionogenicity, while treatment with trypsin or phospholipase C reduced their negative charge.

Bacterial Adhesion↗

Partial characterization of the cohemolytic factor produced by Streptococcus uberis and comparison with the CAMP-factor.

Exosubstances (cohemolysins) produced by Streptococcus agalactiae (CAMP-factor) and Streptococcus uberis (Uberis-factor) showing hemolytic synergism with beta-lysin produced by Staphylococcus aureus were compared. Cohemolytic activity was evaluated in the supernatants of bacterial cultures, before and after ammonium sulfate precipitation. Sheep erythrocytes sensitized with beta-lysin were used as substrate. The assays were performed in microtiter plates and results were expressed as cohemolytic units/ml. Maximum cohemolytic activity was detected, respectively, after 8 h and 14 h of growth in Columbia broth in S. uberis and S. agalactiae cultures. Cohemolytic activities of both microorganisms showed similarities when submitted to various physical and chemical treatments. They were significantly decreased by heating at 60 degrees C and 100 degrees C, or in presence of trypsin, and were abolished in the presence of Tween 20. Activities were found to be stable in crude supernatants and concentrated preparations maintained at -20 degrees C for 3 months. Differences were related to levels of activity and kinetics of detection during the growth cycle. The results indicate the proteic nature, at least in part, of the Uberis factor. Analysis by PAGE in the presence or absence of SDS allowed us to correlate Uberis activity with a protein band with apparent molecular mass of 42 kDa, while CAMP activity was associated with a protein band of 27 kDa.

Bacterial Proteins↗

Correlation between phenotypic characteristics and DNA relatedness within Enterococcus faecium strains.

We noted that a number of enterococcal strains isolated from human clinical specimens resembled Enterococcus faecium but were able to produce acid from glycerol, raffinose, and/or sorbitol, while others failed to form acid from mannitol. An additional concern was that many of these strains with atypical phenotypic characteristics also appeared to acquire vancomycin resistance. In order to determine if such atypical strains were variants of E. faecium or new Enterococcus species, 35 E. faecium or E. faecium-like strains (grouped into 10 phenotypes on the basis of the results of the following tests: capacity to form acid from glycerol, mannitol, raffinose, or sorbitol and susceptibility to vancomycin) and four strains of Enterococcus faecalis were taken from our culture collection, analyzed for their whole-cell protein profiles by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and identified to the species level by DNA-DNA reassociation experiments. All E. faecium-like strains, including four mannitol-negative variants, conformed to at least two of three DNA-DNA relatedness criteria: they were 70% or more related to the type strain of E. faecium at optimal conditions, they had less than 5% divergence within the related sequences, and they had a relatedness of 60% or greater under stringent conditions. The protein profiles of atypical strains were similar to those of typical strains and were easily distinguishable from those of E. faecalis and other enterococcal species. The five E. faecalis strains were 12 to 16% related to the E. faecium type strain. These results indicate that the phenotypic description of E. faecium should include all of these variable characteristics.

Bacterial Proteins↗

Characterization of enterococci isolated from human and nonhuman sources in Brazil.

A total of 330 enterococci strains isolated from several human (272 strains) and animal (27) clinical specimens and environmental sources (31) in Brazil were identified to species level. Major human sources included urine (48.5%), blood (15.8%), and wounds (9.5%). Human isolates were identified as Enterococcus faecalis (90.0%), E. faecium (6.9%), E. gallinarum (1.1%), E. durans (0.8%), E. casseliflavus (0.4%), E. raffinosus (0.4%), and E. mundtii (0.4%). Strains isolated from animals were composed of E. hirae (40.7%), E. faecalis (33.3%), E. faecium (18.5%), and E. casseliflavus (7.5%). Among environmental isolates, 42.0% were E. faecalis, 35.4% E. faecium, 13.0% E. hirae, 6.4% E. casseliflavus, and 3.2% E. durans. Antimicrobial susceptibility tests were performed for 200 strains. Overall, high-level resistance (HLR) to aminoglycosides was found in 66 (33.0%) of the strains tested. HLR to gentamicin was detected in 11.5% of the strains, whereas 19.0% of the strains showed HLR to streptomycin and 26.0% showed HLR to kanamycin. Five (22.7%) of the E. faecium strains were resistant to ampicillin [minimum inhibitory concentration (MIC) > 32 micrograms/ml]. Vancomycin MIC50 and MIC90 were 2 and 4 micrograms/ml, respectively; only eight strains (identified as E. casseliflavus or E. gallinarum) had MIC of 8 micrograms/ml. No beta-lactamase activity was detected by the nitrocefin test.

Animals↗