Multiple-Drug Resistant Nasopharyngeal Streptococcus pneumoniae Isolated in Russia: Serotypes, Antimicrobial Susceptibility, and Molecular Characterization of the Emergent Serotype 13/ST2754 Lineage.

The pneumococcal population structure and drug resistance patterns are constantly changing worldwide. In this study, we described serotypes and antimicrobial susceptibility among 478 multiple-drug resistant (MDR) pediatric nasopharyngeal pneumococci recovered in 2010-2017. The majority of isolates (89.3%; n = 427) carried pneumococcal conjugate vaccine (PCV)13 serotypes, predominantly 6A/B, 14, 19A/F, and 23F. A non-PCV13 serotype capsule was detected in 44 (9.2%) MDR pneumococci, including serotypes 23A (n = 8), 13 (n = 7), 28F (n = 6), 11A (n = 5), and serogroup 35 (n = 10) isolates. The remaining seven (1.5%) MDR isolates were nontypeable. The majority of non-PCV13-serotype isolates were resistant to tetracycline, erythromycin, and clindamycin; most harbored both the ermB and mef genes. Among the 44 serotyped MDR non-PCV13 isolates, multilocus sequence typing analysis revealed 24 different sequence types (STs). ST2754 was the most abundant lineage demonstrating an unusual association with serotypes 13 (n = 7) and 9N (n = 1). The whole-genome sequencing-based analysis demonstrated that the serotype 13/ST2754 lineage was closely related to the serotype 13/ST2754 isolate recovered in Africa (Malawi) in 2013, possessed a Tn6002-like transposon carrying the erm(B) and tet(M) genes, and harbored additional virulence determinants, including arginine metabolism genes and a putative bacteriocin locus. Such a favorable genetic background may provide competitive advantages and potential for spreading and expansion of this clone among pneumococci. These data warrant further molecular monitoring of the genetic composition of the changing pneumococcal population.

Changing serotype distribution and resistance patterns among pediatric nasopharyngeal pneumococci collected in Moscow, 2010-2017.

Serotype distribution and antimicrobial resistance were analyzed in 632 nasopharyngeal pneumococcal isolates collected at a single pediatric center in 2010-2017 before and following the introduction of the 13-valent pneumococcal conjugated vaccine (PCV13) in Russia in 2014. The mean prevalence of PCV13 serotypes was 77.7% in 2010-2015 with a significant decline to 58.5% in 2017, which was accompanied by an elevation in serotype 15B/C prevalence (15.1% in 2017), 66% and 26% of 15B/C-pneumococci related to ST1025 and ST1262, respectively. The rate of oxacillin, erythromycin, and clindamycin resistance has increased by 15-20 percentage points from 2010 to 2016, approaching a 40-45% prevalence in 2016. The resistance rates significantly increased over time only in a group of PCV13 serotypes. The growing resistance among serotype 14 pneumococci was associated with expansion of a multidrug-resistant clone of ST143. These results emphasize the need for close monitoring of the constantly changing pneumococcal population.

Emergence of the Uncommon Clone ST944/ST78 Carrying blaOXA-40-like and blaCTX-M-like Genes Among Carbapenem-Nonsusceptible Acinetobacter baumannii in Moscow, Russia.

Carbapenem-nonsusceptible (Carba-NS) Acinetobacter baumannii has emerged as an important cause of nosocomial infections. In the present study, we characterized 91 Carba-NS A. baumannii isolates collected from patients of surgical departments and intensive care units at three hospitals in Moscow in 2012-2015. Multilocus sequence typing (MLST) using the Oxford (Oxf) scheme identified 16 sequence types (STs) of three clonal complexes (CCs), including CC92Oxf (67%), CC109Oxf (1%), CC944Oxf (29%), and the singleton ST1100Oxf (3%). CC944Oxf was composed of ST944Oxf (n = 16) and two of its newly described single locus variants ST1103Oxf (n = 3) and ST1104Oxf (n = 7); all the three STs were identical to the Pasteur (Pas) MLST scheme ST78. All CC944Oxf/ST78Pas isolates were blaOXA-40-like positive and all but one isolate harbored a blaCTX-M-like gene. ST944Oxf was the only ST found in each of the three study hospitals. Biofilm growth capacity was similar among Carba-NS and nonclonal carbapenem-susceptible isolates. Our data demonstrate the predominance of two clonal lineages among Carba-NS A. baumannii. One of these, the uncommon blaOXA-40-like/blaCTX-M-like-positive clone of CC944Oxf/ST78Pas, seems to be endemic in Russia.

Detection of respiratory pathogens in pediatric acute otitis media by PCR and comparison of findings in the middle ear and nasopharynx.

We conducted a series of polymerase chain reactions (PCRs) in order to detect bacteria (7 species) and viruses (17 species) in middle ear fluid (MEF) and nasopharynx (Nph) of children with acute otitis media (AOM; n=179). Bacterial and viral nucleic acids were detected in MEF of 78.8% and 14.5% patients, respectively. The prevalence was as follows: Streptococcus pneumoniae, 70.4%; Haemophilus influenzae, 17.9%; Staphylococcus aureus, 16.8%; Streptococcus pyogenes, 12.3%; Moraxella catarrhalis, 9.5%; rhinovirus, 9.5%; and adenovirus, 3.4%. The overall rate of PCR-positive specimens for bacterial pathogens was 2.6 times higher, compared to culture results. The rate of PCR-positive results and the distribution of pathogens in the Nph were similar to those in the MEF. Nph PCR results had variable positive predictive values and high negative predictive values in predicting MEF findings. Our results indicate that Nph PCR could be a practical tool for examining respiratory pathogens in children with acute infections.

Bacterial etiology of acute otitis media and characterization of pneumococcal serotypes and genotypes among children in Moscow, Russia.

BACKGROUND: We aimed to describe bacterial etiology of acute otitis media (AOM) and characterize resistance, serotypes and genotype profiles of AOM-causing pneumococci recovered in Moscow children. METHODS: Children with AOM and an available middle ear fluid specimen were prospectively enrolled in this study. Streptococcus pneumoniae, Streptococcus pyogenes, Haemophilus influenzae and Moraxella catarrhalis were considered as true otopathogens. All pneumococcal isolates were serotyped using the Quellung reaction; multidrug-resistant (MDR) pneumococci underwent multilocus sequence typing. RESULTS: In 172 of 541 enrolled AOM patients (32%) at least 1 otopathogen was recovered, with S. pneumoniae having the highest rate of 63% (109/172). When adjusted for antibiotic treatment before sampling, in untreated patients the rate of culture-positive AOM was 35% (124/352), S. pneumoniae had a prevalence of 69% (86/124), S. pyogenes 19% (24/124), H. influenzae 13% (16/124) and M. catarrhalis 9% (11/124). Among 107 examined pneumococci, 45% were penicillin-nonsusceptible, 34 and 30% were resistant to erythromycin and clindamycin, respectively; 30% had an MDR phenotype, but no amoxicillin-resistant isolates were found. Ten of 32 (31%) MDR pneumococci related to clonal complex 320, the remaining isolates belonged to 7 different clonal complex. Six leading serotypes were 19F (27%), 3 (12%), 6B (11%), 14 (11%), 19A (9%) and 23F (8%); overall polysaccharide conjugate vaccine13 coverage was 93%. CONCLUSIONS: S. pneumoniae, the leading bacterial AOM pathogen in Moscow children, is characterized by a substantial rate of antibiotic nonsusceptibility and clonality. A polysaccharide conjugate vaccine with expanded coverage seems to fit the current AOM pneumococcal serotype distribution in Russia better.

Serotypes and antibiotic resistance of non-invasive Streptococcus pneumoniae circulating in pediatric hospitals in Moscow, Russia.

BACKGROUND: Pneumococcal infections remain a major medical problem associated with high morbidity and mortality. Moreover, the resistance of Streptococcus pneumoniae to conventional antibiotics is constantly growing. The implementation of pneumococcal conjugate vaccines (PCVs) in the last decade has dramatically reduced the incidence of the vaccine type-associated invasive pneumococcal diseases in many countries. However, information on the seroepidemiology of S. pneumoniae in Russia is limited. METHODS: We report the results of serotyping and antibiotic susceptibility testing performed on 863 non-invasive pneumococcal isolates collected prospectively in 2009-2013 from children (median age 3.5 years) who sought medical care at five pediatric hospitals in Moscow. The isolates were recovered from the nasopharynx (71.2%), middle ear fluid (14.3%), and lower respiratory tract specimens (13.6%). RESULTS: In total, we identified 45 different serotypes. The six leading serotypes (prevalence >5%) included 19F (21.7%), 6B (12.8%), 23F (10.1%), 14 (9.0%), 6A (8.4%), and 3 (7.5%). Serotype 19A isolates had a prevalence of 2.3%. The proportion of PCV-13 serotypes was 78%; the coverage by PCV-7 was 58.2% and was similar to that of PCV-10 (59.8%). The rate of multidrug-resistant pneumococci (i.e., resistant to ≥3 antimicrobials) was 22%. The majority of the multidrug-resistant isolates were serotype 6B, 14, 19A, and 19F. Penicillin non-susceptibility was displayed by 28% of the isolates. The resistance rate to erythromycin was 26%. Among the examined erythromycin-resistant strains, 54% had the erm(B) gene and 13% had the mef gene as a single resistance determinant, whereas both determinants were found in 31% of these strains. CONCLUSIONS: Our data predict a good coverage of the circulating S. pneumoniae by the PCVs and could be useful for evaluating the serotype distribution in support of the introduction of PCV in Russia. In addition, the antimicrobial resistance rate of S. pneumoniae in Russia is substantial, and the emergence of pneumococcal strains with a dual macrolide resistance mechanism is alarming.