Nathalie van der Mee-Marquet

Identification of Streptococcus agalactiae Isolates from Various Phylogenetic Lineages by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry

ABSTRACT Variations in proteins related to bacterial diversity may affect species identification performed using matrix-assisted laser desorption ionization (MALDI)-time of flight mass spectrometry. Using this method, we identified 110 Streptococcus agalactiae isolates characterized by serotyping and multilocus sequence typing. Serotype III and sequence type 23 strains expressed the widest variation in molecular weight of putative “species-identifying” biomarker ions. Recognition of the diversity of MALDI patterns observed in strains that represent all major intraspecies lineages assists in the constitution of an optimal reference database.

Most Multidrug-Resistant Pseudomonas aeruginosa Isolates from Hospitals in Eastern France Belong to a Few Clonal Types

ABSTRACT This study aimed to determine the genetic diversity of clinical multidrug-resistant Pseudomonas aeruginosa. We used pulsed-field gel electrophoresis and multilocus sequence typing to analyze 187 strains isolated in different French hospitals. To illustrate the diversity of resistance mechanisms to antibiotics in a given clone, we identified β-lactamases with an extended spectrum by using phenotypic and genotypic methods. Typing results showed that the majority of our multidrug-resistant isolates belong to a few clonal types (ST235, ST111, and ST175) that are already spreading worldwide. These successful international clones sporadically produced extended-spectrum β-lactamase-encoding genes but mostly became extensively resistant to β-lactams after derepression of intrinsic resistance mechanisms (i.e., AmpC cephalosporinase). Our results indicate that cross-transmission plays a major role in the spread of multidrug-resistant P. aeruginosa in hospital settings.

Emergence of Unusual Bloodstream Infections Associated with Pig-Borne–Like Staphylococcus aureus ST398 in France

To the Editor—Staphylococcus aureus ST398 is a zoonotic agent primarily described in Europe that is becoming a worldwide threat associated with livestock , their human contacts, and food products. In animals, carriage is frequent , but infections are rare. In humans , infections consist in nosocomial bloodstream and wound infections [1] that are associated with spa types 011 or 034, tetracycline resistance, and the absence of panton-valentine leukocidin (PVL). Recently, a new population of ST398 strains has been isolated in China and from children adopted from China [2] that is responsible for pneumonia and skin and soft-tissue infections in patients without association with animals or animal farming and which is characterized by spa type 571, tetracy-cline susceptibility, and variable presence of PVL [3]. Annual surveys of bloodstream infection are performed in the center region of France [4, 5]. In 2009, we observed the emergence of cases associated with t571, TetS, and PVL-negative ST398 strains. Examination of patient histories revealed exposure to animals in 1 case, a fatal id-iopathic community-acquired bloodstream infection in an 84-year-old man who lived on a farm at which 1 pig was being raised. The remaining cases were hospital-acquired and included 1 case of catheter-associated infection observed in a 58-year-old man with advanced multiple myeloma, 1 case following elective digestive tract surgery in a 69-year-old woman, and 1 case following cardiac surgery in a 68-year-old man. Microarray and MLVA analysis [7] were performed to characterize the strains and compare them with Euro-pean pig-borne methicillin-resistant S. aureus and virulent strains (USA300, MW2, TW20, COL, and Newman). Most characteristics of the present strains were similar to those of the pig-borne strain. All were of accessory gene regulation type I, and none contained the genes encoding the following virulence factors: EsxA and EsxB proteins; leukocidin F; Panton-Valentine Leukocidin; TSST-1; ex-foliatins A and B; and enterotoxins A–E, G–R, and U. None harbored the lantibiotic epidermin/gallidermin genes epiA–epiF typically reported in virulent strains. In addition, similarly to the pig-borne strain, our strains harbored the cna gene, encoding a collagen adhesin associated with colonizing strains and involved in the pathogenesis of osteomyelitis and infectious arthritis [6]; the hyaluronidase gene, involved in the early stages of subcutaneous infections; and a factor SAV2371, associated with bacterial attachment to host cells and virulence. By contrast, unlike the pig-borne strain, the studied strains and the USA300 virulent clone shared cadC–cadM genes, which are responsible for cadmium resistance, …

Methicillin-Susceptible ST398 Staphylococcus aureus Responsible for Bloodstream Infections: An Emerging Human-Adapted Subclone?

In the course of an annual 3-month bloodstream infections (BSI) survey conducted during a four-year period in 31 healthcare institutions located in three noncontiguous French regions, we report 18 ST398 Staphylococcus aureus BSI. ST398 BSI incidence showed a seven-fold increase during the study period (0.002 per 1,000 patient days in 2007 vs. 0.014 in 2010). ST398 BSI isolates differed from the pig-borne multiresistant clone: 17/18 BSI isolates were methicillin susceptible and none was of t011, t034 or t108 pig-borne spa-types. ST398 BSI isolates had homogenous resistance patterns (15/18 with only Eryr) and prophagic content (all harboured the hlb-converting Sau3int phage). The clustering of BSI and pig-borne isolates by spa-typing and MLVA, the occurrence of Sau3int phage in BSI isolates and the lack of this phage in pig-borne isolates suggest that the emergence of BSI isolates could have arisen from horizontal transfer, at least of the Sau3int phage, in genetically diverse MSSA ST398 isolates. The acquisition of the phage likely plays a role in the increasing ability of the lysogenic ST398 isolates to colonize human. The mode of acquisition of the non pig-borne ST398 isolates by our 18 patients remains unclear. ST398 BSI were diagnosed in patients lacking livestock exposure and were significantly associated with digestive portals of entry (3/18 [16.7%] for ST398 vs. 19/767 [2.5%] for non ST398 BSI; p = .012). This raises the question of possible foodborne human infections. We suggest the need for active surveillance to study and control the spread of this human-adapted subclone increasingly isolated in the hospital setting.

Molecular Characterization of Human-Colonizing Streptococcus agalactiae Strains Isolated from Throat, Skin, Anal Margin, and Genital Body Sites

ABSTRACT 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 (STs) were identified. The MLST characteristics of the strains isolated from a single anatomic site—genital (vagina [women] or from a sample of the first urination after arising from a nights sleep [men]), throat, skin, or anal margin—suggest a body site colonization specificity for particular STs: strains of STs 2, 10, 19, and 196 were isolated only from genital sites; strains of STs 1, 8, and 23 were isolated more frequently from throat florae; and strains recovered only from anal margin samples were more closely related to strains isolated from throats than to those from genital sites. Most strains of STs 1, 8, and 23—STs that are increasingly described as being responsible for adult infections—did not carry any markers of strains virulent for neonates, suggesting that the virulence of these strains is probably associated with other genetic determinants. In addition, the genetic diversities of the strains varied between STs: STs 2, 8, 10, 23, and 196 were the most diverse; STs 1 and 19 were more homogeneous; and ST 17 strains formed three distant groups.

Evaluation of the Ability of Streptococcus agalactiae Strains Isolated from Genital and Neonatal Specimens To Bind to Human Fibrinogen and Correlation with Characteristics of the fbsA and fbsB Genes

ABSTRACT The ability of 111 Streptococcus agalactiae strains to bind to human fibrinogen was quantified. We correlated the percentages of bacteria that bound to immobilized fibrinogen with fibrinogen-binding (fbs) gene characteristics of strains and with clinical origin, serotypes, and phylogenetic positions of strains. Percentages varied from 0.4 to 29.9%. Fifty-five strains (49.5%) had the fbsB gene sensu stricto described by Gutekunst et al. (Infect. Immun., 72:3495-3504, 2004), allowing adhesion to human fibrinogen, and all of the other strains had an fgag variant gene. Ninety strains (81.1%) had a fbsA gene and 55 of them also had the fbsB gene. The other 21 strains (18.9%) had a truncated form of fbsA without the fbsB gene sensu stricto. The numbers of 48-nucleotide repeat sequences (rs) in the fbsA gene varied from 2 to 26. The population of strains with the highest ability to bind to human fibrinogen significantly more frequently had the fbsB gene sensu stricto and 4 to 7 rs in the fbsA gene (P < 0.05). However, the single strain that carried the highest number of rs (26 rs) in the fbsA gene showed high fibrinogen-binding activity (24.3%). Strains exhibiting significantly higher levels of binding to human fibrinogen belonged to a phylogenetic group of strains associated with neonatal meningitis, currently known as the ST-17 clone, that is mostly composed of serotype III strains. These findings indicate that S. agalactiae strains possess a wide variety of fbs gene content that markedly influences the ability of strains to bind to human fibrinogen. Variations in the configuration and the expression of the Fbs proteins may therefore partly explain the variability of virulence in S. agalactiae species.

Staphylococcal Exanthematous Disease in a Newborn Due to a Virulent Methicillin-Resistant Staphylococcus aureus Strain Containing the TSST-1 Gene in Europe: an Alert for Neonatologists

ABSTRACT We report a case of staphylococcal exanthematous disease in a newborn due to a toxic shock syndrome toxin 1- and SEC-producing methicillin-resistant Staphylococcus aureus strain and alert neonatologists to the probable emergence in France of the neonatal toxic shock syndrome-like exanthematous disease in newborns previously described in Japan. We advise neonatologists to pay careful attention to clinical parameters and to prescribe appropriate tests: platelet count, serum C-reactive protein concentration, and Vβ2-positive T-cell counts.

Molecular characterization of erythromycin-resistant Streptococcus agalactiae strains

OBJECTIVES The aim of this study is to identify the molecular characteristics of erythromycin-resistant (Erm(r)) Streptococcus agalactiae strains and to correlate with the clinical origin of strains. METHODS From 711 S. agalactiae strains, 119 Erm(r) strains (17%) were collected, serotyped and screened for macrolide resistance genes. The genetic relationship between strains was established by the PFGE analysis. Strains were tested for the group II intron GBSi1 downstream of the scpB gene, IS1548 in the hylB gene, four prophage DNA fragments and a lineage defined by multilocus sequence typing as ST-17. RESULTS Erythromycin resistance involved 8% of serotype Ia, 15% of serotype Ib, 9% of serotype II, 16% of serotype III, 31% of serotype IV and 35% of serotype V. The prevalence of Erm(r) strains was higher among strains isolated from the gastric fluid of neonates (33%) than in those isolated from bacteraemia and meningitis during early-onset disease (EOD) or late-onset disease (7% and 11%) (P = 0.001). In serotype III, Erm(r) strains were more frequent in vaginal carriage (22%) and colonized neonates (18%) than in EOD (0%) (P = 0.03). The mef(A) gene was the most common in serotype Ia (55%), the erm(A) gene in serotype Ib (75%) and the erm(B) gene in the other serotypes (56% to 75%). All resistant strains with IS1548 also had the erm(B) gene. Erm(r) strains were not randomly distributed in the different PFGE genogroups, and 11% had the GBSi1 intron, 37% had at least one prophage DNA fragment and 7% belonged to ST-17. CONCLUSIONS Erythromycin resistance varied according to the clinical origin, serotype and molecular characteristics of S. agalactiae strains.

Legionella anisa, a Possible Indicator of Water Contamination by Legionella pneumophila

ABSTRACT Legionella anisa is one of the most frequent species of Legionella other than Legionella pneumophila in the environment and may be hospital acquired in rare cases. We found that L. anisa may mask water contamination by L. pneumophila, suggesting that there is a risk of L. pneumophila infection in immunocompromised patients if water is found to be contaminated with Legionella species other than L. pneumophila.

Acquisition of Insertion Sequences and the GBSi1 Intron by Streptococcus agalactiae Isolates Correlates with the Evolution of the Species

The prevalence of insertion sequences IS1548, IS861, IS1381, and ISSa4 and of the group II intron GBSi1 within Streptococcus agalactiae human isolates strongly correlates with the genetic lineages obtained by multilocus sequence typing. Our results yielded an evolutionary scheme for the acquisition of these genetic elements linked to the ecosystems from which the isolates were obtained.