High rate of fecal carriage of extended-spectrum-ß-lactamase-producing Escherichia coli in healthy children in Gipuzkoa, northern Spain.

The prevalence of extended-spectrum-ß-lactamase-producing Enterobacteriaceae (ESBLPE) was studied in stool samples from 125 8- to 16-month-old healthy children. Twenty-four percent of them and 10.7% of the 318 fecal samples studied yielded extended-spectrum-ß-lactamase-producing Escherichia coli, with the types being SHV-12, CTX-M-1, CTX-M-14, and TEM-52, the most common types of ß-lactamases. This high prevalence of ESBLPE in healthy people, which is to our knowledge the highest currently reported in Europe, may represent a risk for increased infections by these organisms in the future.

What causes decreased erythromycin resistance in Streptococcus pyogenes? Dynamics of four clones in a southern European region from 2005 to 2012.

OBJECTIVES: To survey antibiotic resistance among Streptococcus pyogenes isolates collected from 2005 to 2012, to characterize those showing erythromycin resistance and to analyse the association of certain emm types with erythromycin resistance or susceptibility. METHODS: Resistance determinants or mutations conferring erythromycin, clindamycin, tetracycline and fluoroquinolone resistance were analysed. All erythromycin-resistant isolates and a sample of erythromycin-susceptible isolates were emm typed. Multilocus sequence typing was performed for representative emm types. RESULTS: Antimicrobial susceptibility was studied for 12 346 S. pyogenes isolates. Erythromycin, clindamycin and tetracycline resistance showed a decreasing trend. In 2012, 2.8% of isolates were erythromycin resistant versus 7.5% in 2005 and 11.7% in 2006. Although 21 clones were involved, 4 clones accounted for almost 90% of erythromycin-resistant isolates. The emm12/ST36 clone, carrying the mef(A) gene, was the predominant (41.1%) erythromycin-resistant clone, with an incidence peak in 2008, followed by a gradual decline. The M phenotype predominated each year except for 2005, when two of the main erythromycin-resistant clones (emm11/ST403 and emm28/ST52) harboured an erm(B) gene. Erythromycin resistance was significantly higher in adults than in children. Skin isolates showed the highest erythromycin resistance rate; among these, perianal isolates frequently belonged to the emm28/ST52 clone. The emm type was not a predictor of erythromycin resistance; however, most emm11 and emm12 were erythromycin-resistant isolates. Macrolide consumption was similar throughout the study period. Only two isolates with a high level of levofloxacin resistance were detected. CONCLUSIONS: Resistance was mainly related to the circulation of emm12/ST36, emm11/ST403, emm28/ST52 and emm4/ST39 clones, all of which declined throughout the study period.

Prevalence of genosubtypes (PorA types) of serogroup B invasive meningococcus in the north of Spain from 2000 to 2003.

The composition of new vaccines against Neisseria meningitidis serogroup B is based on differences in the variable regions VR1 and VR2 of the class 1 outer-membrane protein (PorA) of meningococci. Genosubtyping of 96 N. meningitidis B isolates from blood or cerebrospinal fluid from 2000 to 2003 in the north of Spain allowed characterization of all the strains. Twenty-six genosubtypes or distinct PorA types were obtained. The most prevalent were P1.5-1, 10-8 (20 strains), P1.19, 15 (14 strains), P1.22, 9 (11 strains) and P1.5, 2 (nine strains), while 17 genosubtypes were represented by only one or two strains. The wide diversity of genosubtypes observed and their differences compared with those found in other regions reveals the difficulty in designing a useful outer-membrane vesicle vaccine applicable to different regions of the world.

Progressive decrease in the potential usefulness of meningococcal serogroup B vaccine (4CMenB, Bexsero®) in Gipuzkoa, Northern Spain.

The effectiveness of a vaccine is determined not only by the immunogenicity of its components, but especially by how widely it covers the disease-causing strains circulating in a given region. Because vaccine coverage varies over time, this study aimed to detect possible changes that could affect vaccine protection during a specific period in a southern European region. The 4CMenB vaccine is licensed for use in Europe, Canada, and Australia and is mainly directed against Neisseria meningitidis serogroup B. This vaccine contains four main immunogenic components: three recombinant proteins, FHbp, Nhba and NadA, and an outer membrane vesicle [PorA P1.4]. The allelic distribution of FHbp, Nhba, NadA, and PorA antigens in 82 invasive isolates (B and non-B serogroups) isolated from January 2008 to December 2013 were analyzed. 4CMenB was likely protective against 61.8% and 50% of serogroup B and non-B meningococci, respectively, in the entire period, but between 2012 and 2013, the predicted protection fell below 45% (42.1% for serogroup B isolates).The observed decreasing trend in the predicted protection during the 6 years of the study (Χ2 for trend  = 4.68, p = 0.03) coincided with a progressive decrease of several clonal complexes (e.g., cc11, cc32 and cc41/44), which had one or more antigens against which the vaccine would offer protection.

Fatal Neisseria meningitidis serogroup X sepsis in immunocompromised patients in Spain. Virulence of clinical isolates.

BACKGROUND: Neisseria meningitidis X (NmX) is a minority meningococcus serogroup for which no vaccine is available. Huge meningococcus X outbreaks occurred in West Africa but developed countries have reported only sporadic cases. Two invasive fatal cases are presented. METHODS: Isolates were characterized by serogrouping (latex agglutination and genogroup X-specific polymerase-chain-reaction), PorA and FetA typing, multilocus sequence typing and pulsed-field gel electrophoresis. Immunocompetent female balm/C mice inoculated intraperitoneally were use to test virulence of invasive and carrier isolates. RESULTS: Until April 2010, NmX was absent among 868 invasive meningococci characterized in the Basque Country in a 20-year period. In April 2010, two fatal NmX: P1.21,16: F5-5/ST750 episodes were detected in unrelated immunodeficient patients. After analysis of 803 meningococcal isolates from asymptomatic carriers obtained between 1988 and 2010, eight NmX isolates were detected. The genotype of the two invasive isolates bore no relation to any of the NmX isolates detected in healthy individuals from the Basque Country or to isolates from outbreaks in Africa. CONCLUSIONS: NmX isolates in the north of Spain can cause severe disease in humans, despite their low prevalence. The in vivo animal study showed that virulence of isolates was more closely associated with the genotype than with the serogroup.

Influence of two vaccination campaigns on genetic diversity of invasive Neisseria meningitidis isolates in northern Spain (1997-2008).

BACKGROUND: Neisseria meningitidis diversifies rapidly, due to its high recombination rates. The aim of this study was to analyze the possible impact of two vaccination campaigns (a once-off A/C polysaccharide vaccination campaign in people aged 18 months to 20 years old in 1997, and a meningococcal C conjugate vaccination campaign in children aged < or = 6 years old from 2000 to 2008) on diversification of the population of invasive isolates obtained between 1997 and 2008. All of the 461 available isolates were included (2, 319, 123, 11 and 6 belonging to serogroups A, B, C, Y and W-135, respectively). METHODOLOGY/PRINCIPAL FINDINGS: The isolates were analyzed for diversity using multilocus sequence typing, eBURST and the S.T.A.R.T.2 program. One hundred and seven sequence types (ST) and 20 clonal complexes were obtained. Five different STs (ST11, ST8, ST33, ST1163 and ST3496) included 56.4% of the isolates. With the exception of ST11, all other STs were associated with a specific serogroup. Epidemic circulation of serogroup C ST8 isolates was detected in 1997-1998, as well as epidemic circulation of ST11 isolates (serogroups B and C) in 2002-2004. The epidemic behavior of serogroup B ST11 (ST11_B:2a:P1.5) was similar, although with lesser intensity, to that of ST11 of serogroup C. Although clonality increased during epidemic years, the overall diversity of the meningococcal population did not increase throughout the 12 years of the study. CONCLUSION: The overall diversity of the meningococcal population, measured by the frequency of STs and clonal complexes, numbers of alleles, polymorphic sites, and index of association, remained relatively constant throughout the study period, contradicting previous findings by other researchers.