RESUMO
In the present study, 37 group A Streptococcus (GAS) strains belonging to 13 new emm sequence types identified among GAS strains randomly isolated in Brazil were characterized by using phenotypic and genotypic methods. The new types were designated st204, st211, st213, st809, st833, st854, st2904, st2911, st2917, st2926, st3757, st3765, and st6735. All isolates were susceptible to the antimicrobial agents tested, except to tetracycline. They all carried the speB gene, and 94.6% produced detectable SpeB. Most strains belonging to a given emm type had similar or highly related pulsed-field gel electrophoresis profiles that were distinct from profiles of strains of another type. The other characteristics were variable from isolate to isolate, although some associations were consistently found within some emm types. Unlike the other isolates, all type st213 isolates were speA positive and produced SpeA. Strains belonging to st3765 were T6 and opacity factor (OF) negative. Individual isolates within OF-positive emm types were associated with unique sof gene sequence types, while OF-negative isolates were sof negative by PCR. This report provides information on new emm sequence types first detected in GAS isolates from a geographic area not extensively surveyed. Such data can contribute to a better understanding of the local and global dynamics of GAS populations and of the epidemiological aspects of GAS infections occurring in tropical regions.
Assuntos
Antígenos de Bactérias , Proteínas da Membrana Bacteriana Externa/genética , Proteínas de Transporte/genética , Infecções Estreptocócicas/microbiologia , Streptococcus pyogenes/classificação , Proteínas da Membrana Bacteriana Externa/metabolismo , Técnicas de Tipagem Bacteriana , Brasil/epidemiologia , Proteínas de Transporte/metabolismo , Eletroforese em Gel de Campo Pulsado , Genótipo , Humanos , Dados de Sequência Molecular , Fenótipo , Reação em Cadeia da Polimerase , Análise de Sequência de DNA , Infecções Estreptocócicas/epidemiologia , Streptococcus pyogenes/genética , Streptococcus pyogenes/isolamento & purificaçãoRESUMO
All clinical S. pneumoniae specimens isolated from patients with invasive or sterile-site infections admitted to one regional general hospital in southern Chile were collected during a 5-year period (February 1994 to September 1999). A total of 247 strains belonging to 50 serotypes were isolated in this survey: 69 in patients under 5 years of age, 129 in patients 5 to 64 years old, and 49 from patients 65 years and older. Eight serotypes were identified in all age groups, while all other serotypes were found exclusively in one age group or in patients over 4 years of age. Serotype 3 was never found in patients under 5 years old, and serotype 14 was not found in patients >64 years of age. There was no difference in the serotypes causing infection in each one of the 5 years of the survey. Our results suggest that both bacterial virulence factors and host factors play an important role in the selection of S. pneumoniae serotypes causing invasive infection. Possible host factors include age-related differences in the immune response. Comparative studies with other areas of the world may help to further understanding of our observations in southern Chile.
Assuntos
Infecções Pneumocócicas/microbiologia , Streptococcus pneumoniae/patogenicidade , Adolescente , Adulto , Fatores Etários , Criança , Pré-Escolar , Humanos , Pessoa de Meia-Idade , Infecções Pneumocócicas/fisiopatologia , Streptococcus pneumoniae/genética , VirulênciaRESUMO
Seventy-two strains of pediococci isolated from human clinical sources were characterized by conventional physiological tests, chromogenic enzymatic tests, analysis of whole-cell protein profiles (WCPP) by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and analysis of chromosomal DNA restriction profiles by pulsed-field gel electrophoresis (PFGE). Conventional tests allowed identification of 67 isolates: 52 strains were identified as Pediococcus acidilactici, 15 strains were identified as Pediococcus pentosaceus, and 5 strains were not identified because of atypical reactions. Analysis of WCPP identified all isolates since each species had a unique WCPP. By the WCPP method, the atypical strains were identified as P. acidilactici (two strains) and P. pentosaceus (three strains). The chromogenic substrate test with o-nitrophenyl-beta-D-glucopyranoside differentiated all 54 strains of P. acidilactici (negative reactions) and 13 (72%) of 18 strains of P. pentosaceus (positive reactions). Isolates of both species were shown to be nonclonal as revealed by the genetic diversity when chromosomal DNA was analyzed by PFGE. Using WCPP as the definitive identification procedure, P. acidilactici (28 of 54 strains; 51.8%) was more likely than P. pentosaceus (4 of 18 strains; 22.3%) to be isolated from blood cultures.
Assuntos
Técnicas de Tipagem Bacteriana , Infecções por Bactérias Gram-Positivas/microbiologia , Pediococcus/classificação , Pediococcus/genética , Compostos Cromogênicos/metabolismo , DNA Bacteriano/análise , Eletroforese em Gel de Campo Pulsado , Eletroforese em Gel de Poliacrilamida , Genótipo , Humanos , Pediococcus/isolamento & purificação , Pediococcus/fisiologia , FenótipoRESUMO
BACKGROUND: Outbreaks of nephritis have been rare since the 1970s. From December, 1997, to July, 1998, 253 cases of acute nephritis were identified in Nova Serrana, Brazil. Seven patients required dialysis, and three patients died. We did a case-control study to investigate the cause of the outbreak. METHODS: Using a matched cluster design, we examined seven recent patients, their family members (n=23), and members of neighbourhood-matched control households (n=22). We subsequently interviewed 50 patients and 50 matched controls about exposure to various dairy products. We also cultured dairy foods and took udder-swab and milk samples from cows. FINDINGS: Throat cultures indicated that nephritis was associated with group C Streptococcus equi subspecies zooepidemicus, a cause of bovine mastitis. S. zooepidemicus was detected in four of seven case households (six of 30 people) and no control households (p=0.09). Patients were more likely than matched controls to have consumed a locally produced cheese called queijo fresco (matched odds ratio 2.1, p=0.05). The nephritis attack rate was 4.5 per 1000 in Nova Serrana but 18 per 1000 in the village Quilombo do Gaia (p=0.003). The largest supplier of unpasteurized queijo fresco was a farm in Quilombo do Gaia. S. zooepidemicus was not detected in food samples or in swabs collected from cows in August, 1998, although mastitis was evident among cows on the suspected farm. Throat cultures of the two women who prepared cheese on this farm yielded the outbreak strain of S. zooepidemicus. After the cheese was removed from the distribution system, no further cases were reported. INTERPRETATION: A large outbreak of glomerulonephritis was attributed to S. zooepidemicus in unpasteurised cheese. This outbreak highlights the dangers of consuming unpasteurized dairy products and need for global efforts to promote food safety.
Assuntos
Surtos de Doenças , Nefrite/epidemiologia , Infecções Estreptocócicas/epidemiologia , Streptococcus equi/isolamento & purificação , Adolescente , Adulto , Animais , Brasil/epidemiologia , Estudos de Casos e Controles , Bovinos , Feminino , Microbiologia de Alimentos , Humanos , Masculino , Nefrite/microbiologia , Infecções Estreptocócicas/microbiologiaRESUMO
To assess pneumococcal strain variability among young asymptomatic carriers in Chile, we used serotyping, antibiotic susceptibility testing, and genotyping to analyze 68 multidrug-resistant pneumococcal isolates recovered from 54 asymptomatic children 6 to 48 months of age. The isolates represented capsular serotypes 19F (43 isolates), 14 (14 isolates), 23F (7 isolates), 6B (3 isolates), and 6A (1 isolate). Genotypic analysis, which included pulsed-field gel electrophoresis (PFGE) of chromosomal digests, penicillin binding protein (PBP) gene fingerprinting, and dhf gene fingerprinting, revealed that the isolates represented six different genetic lineages. Clear circumstantial evidence of capsular switching was seen within each of four of the genetically related sets. The majority of the isolates, consisting of the 43 19F isolates and 2 type 6B isolates, appeared to represent a genetically highly related set distinct from previously characterized pneumococcal strains. Each of three other genetically defined lineages was closely related to one of the previously characterized clones Spain(6B)-2, France(9V)-3, or Spain(23F)-1. A fifth lineage was comprised of four type 23F isolates that, by the techniques used for this study, were genetically indistinguishable from three recent type 19F sterile-site isolates from the United States. Finally, a sixth lineage was represented by a single type 23F isolate which had a unique PFGE type and unique PBP and dhf gene fingerprints.
Assuntos
Portador Sadio/microbiologia , Infecções Pneumocócicas/microbiologia , Streptococcus pneumoniae/efeitos dos fármacos , Streptococcus pneumoniae/genética , Resistência beta-Lactâmica/genética , Sequência de Bases , Pré-Escolar , Chile , Primers do DNA/genética , Eletroforese em Gel de Campo Pulsado , Variação Genética , Genótipo , Humanos , Lactente , Testes de Sensibilidade Microbiana , Nasofaringe/microbiologia , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , Sorotipagem , Streptococcus pneumoniae/classificaçãoRESUMO
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.
Assuntos
Infecções Estreptocócicas/microbiologia , Streptococcus/classificação , Streptococcus/genética , Animais , Eletroforese , Enzimas/genética , Humanos , Hibridização de Ácido Nucleico , Fenótipo , Filogenia , Sorotipagem , Especificidade da Espécie , Streptococcus/isolamento & purificaçãoRESUMO
Until recently, group B streptococcus, serotype V (GBS-V), was an infrequent cause of disease. It is now recognized as a significant cause of infections in both children and adults. To determine if this increase was due to the recent introduction and spread of a single clone of GBS-V, we analyzed, by pulsed-field gel electrophoresis (PFGE), the SmaI chromosomal DNA digests of 45 bacteria: 41 isolated from human infections between 1986 and 1996 in the United States, 2 from human infections in Argentina, and 2 from naturally infected mice. Seventeen patterns were found and arbitrarily designated patterns A to Q. Pattern N constituted 24 (53%) of the isolates and was found in all of the years tested and from all surveillance areas, as well as in both isolates from Argentina, and was very similar to the GBS-V isolated from a mouse. Pattern P was found in three isolates, pattern F was found in two, and the remaining patterns were found in one isolate each. We concluded that the majority of isolates of GBS-V are of one PFGE subtype and that this subtype was predominate before the increase in disease caused by GBS-V and that GBS-V disease is caused by several different subtypes.
Assuntos
Eletroforese em Gel de Campo Pulsado , Infecções Estreptocócicas/microbiologia , Streptococcus agalactiae/classificação , Adulto , Animais , Argentina/epidemiologia , Técnicas de Tipagem Bacteriana , Criança , DNA Bacteriano/análise , Humanos , Camundongos , Sorotipagem , Infecções Estreptocócicas/epidemiologia , Streptococcus agalactiae/genética , Streptococcus agalactiae/isolamento & purificação , Estados Unidos/epidemiologiaRESUMO
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.
Assuntos
Antibacterianos/farmacologia , Técnicas de Tipagem Bacteriana , Enterococcus/classificação , Metilglucosídeos/metabolismo , Resistência Microbiana a Medicamentos , Enterococcus/efeitos dos fármacos , Enterococcus/metabolismo , Humanos , Testes de Sensibilidade Microbiana , Piridonas/farmacologia , Especificidade da EspécieRESUMO
We compared the incidence of nasopharyngeal colonization by Streptococcus pneumoniae, the serotypes causing mucosal and invasive diseases, and the antibiotic resistance of these strains in patients admitted to three large hospitals and children attending day care centers in two Chilean cities (Santiago and Temuco). The populations in both cities were similar in ethnic background, socioeconomic status, family size, and access to medical care. Significant differences in nasopharyngeal colonization rates, in serotypes causing infections, and in antibiotic resistance were found between the two cities. In children 0 to 2 years of age, 42% were colonized with S. pneumoniae in Santiago compared to 14% in Temuco. A total of 41 serotypes were identified in both Chilean cities studied. Six serotypes were found only in Santiago; 14 serotypes were found only in Temuco. Antibiotic-resistant serotypes 6A, 6B, 14, 19F, and 23F were detected only in Santiago. We show that important differences in the incidence of nasopharyngeal carriage, infection, and S. pneumoniae serotypes can exist in similar populations in different areas of the same country. Our findings are relevant for prevention strategies, antibiotic usage, and vaccine design.
Assuntos
Resistência Microbiana a Medicamentos/genética , Infecções Pneumocócicas/microbiologia , Streptococcus pneumoniae/isolamento & purificação , Adolescente , Criança , Pré-Escolar , Chile/epidemiologia , Humanos , Lactente , Infecções Pneumocócicas/epidemiologia , Sorotipagem , Streptococcus pneumoniae/efeitos dos fármacos , Streptococcus pneumoniae/genéticaRESUMO
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.