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Streptococcus agalactiae carriage was evaluated by sampling four body sites in a group of 249 healthy individuals including both sexes and a wide range of ages; the aims were to study the population structure of colonizing strains by multilocus sequence typing (MLST) and to evaluate their diversity by serotyping, SmaI macrorestriction analysis, and PCR screening for genetic markers of highly virulent clones for neonates. The prevalences of carriage were 27% in women and 32% in men. The major positive body site was the genital tract (23% in women and 21% in men); skin, throats, and anal margins were also positive in 2%, 4%, and 14%, respectively. These human-colonizing strains belonged mostly to serotypes III (24%), Ia (21%), V (18%), and Ib (17%). Twenty-three sequence types ( Streptococcus agalactiae is the pathogen most commonly responsible for maternofetal and neonatal infections, and serogroup III sequence type (ST) 17 strains of this species have been identified as more likely to cause these infections (3,17,(24)(25)(26)28).S. agalactiae infections in nonpregnant adults have been reported increasingly since the 1970s (5,7,10,14,20,21,23,26,27). The major clinical forms consist of skin, soft tissue, and bone infections; bacteremia; urinary tract infections; pneumonia; and peritonitis. More rarely, S. agalactiae is responsible for septic arthritis, meningitis, and endocarditis. Risk factors have been identified (16) and include an age of over 60 years and diagnoses of diabetes, cancer, decubitus acutus, AIDS, longcourse corticotherapy, chronic renal disease, cirrhosis, and neurological vessel disorders. However, cases of invasive S. agalactiae infections in immunocompetent nonpregnant adults have recently been reported (7,20,21,23). The physiopathology of these infections is not well understood and may be linked in part to S. agalactiae strains with particular properties.Reports on the genetic characteristics of the strains associated with infections in adults showed that they are diverse and belong mostly to serotypes Ia, III, and V (5, 14) and clonal complexes 1, 9, 17, 19, and 23 (17).The portal of entry of these infections in adults is not often well documented. A better knowledge of the genetic characteristics of the S. agalactiae strains colonizing various body sites would contribute to a better understanding of the origin of infections in adults. Although the molecular features of S. agalactiae strains isolated during infectious diseases have been described in many cases, little is known about the molecular characteristics of human-colonizing strains. Therefore, we screened for S. agalactiae carriage in 249 healthy individuals of both sexes; the individuals were living at home and covered a range of ages, from the start of the teenage period until old age. Genital, skin, throat, and anal margin carriage was sampled by self-swabbing. The strains isolated were serotyped. The population structure of the strains was investigated by multilocus sequence typing (MLST). The genetic diversity of isolates was ev...

Group B streptococcus (GBS) strains with the highest ability to bind to human fibrinogen belong to the highly invasive clonal complex (CC) 17. To investigate the fibrinogen-binding mechanisms of CC17 strains, we determined the prevalence of fibrinogen-binding genes (fbsA and fbsB), and fbs regulator genes (rogB encoding an fbsA activator, rovS encoding an fbsA repressor and rgf encoding a two-component system [TCS] whose role on fbs genes was not determined yet) in a collection of 134 strains representing the major CCs of the species. We showed that specific gene combinations were related to particular CCs; only CC17 strains contained the fbsA, fbsB, and rgf genes combination. Non polar rgfAC deletion mutants of three CC17 serotype III strains were constructed. They showed a 3.2- to 5.1-fold increase of fbsA transcripts, a 4.8- to 6.7-fold decrease of fbsB transcripts, and a 52% to 68% decreased fibrinogen-binding ability, demonstrating that the RgfA/RgfC TCS inhibits the fbsA gene and activates the fbsB gene. The relative contribution of the two fbs genes in fibrinogen-binding ability was determined by constructing isogenic fbsA, fbsB, deletion mutants of the three CC17 strains. The ability to bind to fibrinogen was reduced by 49% to 57% in ΔfbsA mutants, and by 78% to 80% in ΔfbsB mutants, suggesting that FbsB protein plays a greater role in the fibrinogen-binding ability of CC17 strains. Moreover, the relative transcription level of fbsB gene was 9.2- to 12.7-fold higher than that of fbsA gene for the three wild type strains. Fibrinogen-binding ability could be restored by plasmid-mediated expression of rgfAC, fbsA, and fbsB genes in the corresponding deletion mutants. Thus, our results demonstrate that a specific combination of fbs genes and fbs regulator genes account for the high fibrinogen-binding ability of CC17 strains that may participate to their enhanced invasiveness for neonates as compared to strains of other CCs.

The phylogenetic position and prophage DNA content of the genomes of 142 S. agalactiae (group-B streptococcus, GBS) isolates responsible for bacteremia and meningitis in adults and neonates were studied and compared. The distribution of the invasive isolates between the various serotypes, sequence types (STs) and clonal complexes (CCs) differed significantly between adult and neonatal isolates. Use of the neighbor-net algorithm with the PHI test revealed evidence for recombination in the population studied (PHI, P = 2.01×10−6), and the recombination-mutation ratio (R/M) was 6∶7. Nevertheless, the estimated R/M ratio differed between CCs. Analysis of the prophage DNA regions of the genomes of the isolates assigned 90% of the isolates to five major prophage DNA groups: A to E. The mean number of prophage DNA fragments amplified per isolate varied from 2.6 for the isolates of prophage DNA group E to 4.0 for the isolates of prophage DNA group C. The isolates from adults and neonates with invasive diseases were distributed differently between the various prophage DNA groups (P<0.00001). Group C prophage DNA fragments were found in 52% of adult invasive isolates, whereas 74% of neonatal invasive isolates had prophage DNA fragments of groups A and B. Differences in prophage DNA content were also found between serotypes, STs and CCs (P<0.00001). All the ST-1 and CC1 isolates, mostly of serotype V, belonged to the prophage DNA group C, whereas 84% of the ST-17 and CC17 isolates, all of serotype III, belonged to prophage DNA groups A and B. These data indicate that the transduction mechanisms, i.e., gene transfer from one bacterium to another by a bacteriophage, underlying genetic recombination in S. agalactiae species, are specific to each intraspecies lineage and population of strains responsible for invasive diseases in adults and neonates.