Pneumococcal Strains Research Articles

Dynamics of resistance of Streptococcus pneumoniae strains to antibacterial drugs for the period 2015–2019 in Samara: opportunities for overcoming

According to the World Health Organization (WHO), diseases caused by Streptococcus pneumoniae are one of the most dangerous diseases, that can be completely prevented by vaccination. Studies conducted in various regions of the Russian Federation and other countries have revealed certain features in the prevalence of pneumococcus strains, resistant to different antibacterial drugs, which requires local studies among patients of different age groups.The aim of the study was to determine the level of sensitivity of S. pneumoniae, isolated from adult patients, to antibacterial drugs (ABD) and the dynamics of resistance to ABD in Samara for the period of 2015–2019.Methods. A retrospective analysis of data from the microbiological department of clinical diagnostic laboratories of the Clinics of Samara State Medical University, Healthcare Ministry of Russia for 2015–2019 was carried out. The microorganism resistance to several groups of ABD was studied – macrolides (erythromycin, azithromycin, clarithro mycin), cephalosporins (ceftriaxone, cefotaxime), fluoroquinolones (levofloxacin) and penicillin. The sensitivity was determined by the disk diffusion method. Statistical processing was performed using the SPSS Statistic 22.0 program. When comparing groups, the χ2 criterion was used; to compare small groups (n < 5), the exact Fisher test was used.Results. Since 2017, there has been a tendency to increase the frequency of pneumococcus isolation in the carrier group. At the same time, a 3-fold decrease in S. pneumoniae excretion from patients with ENT pathology occurred. The highest sensitivity was detected for cephalosporins – 100% sensitivity of all S. pneumoniae strains, detected from ENT organs and during carriage. There is a low level of pneumococcus resistance to levofloxacin. The general sensitivity to ABD from penicillin group is 96.4–98.8%, while the most common resistant strains were found in patients with respiratory tract damage. The highest resistance of S. pneumoniae was revealed to ABD from the macrolide group with a tendency to increase from 4.6% in 2016 to 14.0% in 2019.Conclusion. According to the study results, a rather low level of resistance of S. pneumoniae to most ABD groups, with the exception of macrolides, was demonstrated in Samara. The greatest number of ABD-insensitive pathogens was isolated in patients with pathology of the bronchopulmonary system. A mandatory microbiological examination of the sputum of such patients is necessary with a determination of the sensitivity of the isolated microorganisms to ABD and subsequent correction of therapy upon the identification of resistant strains. Given the increasing resistance of S. pneumoniae in carriers, it is necessary to vaccinate patients in this group.

Nasopharyngeal Colonization and levofloxacin Susceptibility of Streptococcus pneumoniae among Healthcare Workers in a Teaching Hospital in Tehran, Iran

Background: Streptococcus pneumoniae commonly asymptomatically colonizes the human upper respiratory tract and the carriage rate varies between geographical regions. The colonized individuals are not only at risk of infections but also can be a source of transmission of the pathogen. The risk of the droplet or airborne transmission of pneumococcal strains healthcare workers is considerable. The current study aimed to determine the extent of nasopharyngeal colonization with S. pneumoniae and their levofloxacin susceptibility at a tertiary hospital in Tehran, Iran. Materials and Methods: During a six-month period, the nasopharyngeal swab samples collected from 300 volunteer healthcare workers of Imam Hossein Hospital. Samples screened for S. pneumoniae using standard conventional biochemical methods. The minimum inhibitory concentration (MIC) of levofloxacin was determined using a commercially available strip antibiotic test according to the clinical laboratory standards institute (CLSI) guidelines. Results: A total of 19 (6.3%) enrolled healthcare workers were colonized with S. pneumoniae. Amongst the enrolled volunteers, nurses had a higher rate of pneumococcal colonization (47.3%) followed by interns (21%) and laboratory workers (15.8%). Our analysis revealed that there was a significant correlation between smoking and pneumococcal colonization. The antimicrobial susceptibility testing showed that all of the isolates were susceptible to levofloxacin (MIC≤2 μg/ml). Conclusion: This low rate of pneumococcal colonization amongst healthcare professionals may be attributed to the low risk of horizontal transmission of severe pneumococcal infections in the hospital. Additionally, our findings indicated that levofloxacin was an effective antimicrobial agent for the treatment of pneumococcal infections.

Adenosine Triphosphate Neutralizes Pneumolysin-Induced Neutrophil Activation

In tissue infections, adenosine triphosphate (ATP) is released into extracellular space and contributes to purinergic chemotaxis. Neutrophils are important players in bacterial clearance and are recruited to the site of tissue infections. Pneumococcal infections can lead to uncontrolled hyperinflammation of the tissue along with substantial tissue damage through excessive neutrophil activation and uncontrolled granule release. We aimed to investigate the role of ATP in neutrophil response to pneumococcal infections. Primary human neutrophils were exposed to the pneumococcal strain TIGR4 and its pneumolysin-deficient mutant or directly to different concentrations of recombinant pneumolysin. Neutrophil activation was assessed by measurement of secreted azurophilic granule protein resistin and profiling of the secretome, using mass spectrometry. Pneumococci are potent inducers of neutrophil degranulation. Pneumolysin was identified as a major trigger of neutrophil activation. This process is partially lysis independent and inhibited by ATP. Pneumolysin and ATP interact with each other in the extracellular space leading to reduced neutrophil activation. Proteome analyses of the neutrophil secretome confirmed that ATP inhibits pneumolysin-dependent neutrophil activation. Our findings suggest that despite its cytolytic activity, pneumolysin serves as a potent neutrophil activating factor. Extracellular ATP mitigates pneumolysin-induced neutrophil activation.

Disruption of Aspergillus fumigatus biofilm by Streptococcus pneumoniae: Mycelial fragmentation by hydrogen peroxide

Biofilm is a complex structure consisting of microorganisms such as bacteria, fungi and an extracellular matrix (ECM). Biofilms are involved in most microbial infections and show persistent resistance to antibiotic treatment and immune response. Both Aspergillus fumigatus and Streptococcus pneumoniae are colonizers that can form biofilms in the respiratory tract. These pathogens have been simultaneously isolated from the same patient, but their interaction is poorly understood. We observed morphological changes in single- and mixed-species biofilms prepared for confocal laser scanning microscopy and scanning electron microscopy (SEM). Pneumococci suppressed the development of a fungal biofilm, and it even disrupted a preformed fungal biofilm. When a preformed fungal biofilm was treated with pneumococci, the mycelial network was fragmented, and only bacteria could develop. SEM revealed that the fragmented mycelium was further disrupted into fine filaments as treatment time progressed, and that the ECM of the preformed fungal biofilm had disappeared. The pneumococcal culture supernatant contained mycelial fragmentation activity that was heat-sensitive. The culture supernatant of a mutant pneumococcal strain deficient in pneumolysin (Δply) also exhibited the mycelial fragmentation activity. Enolase and lactate oxidase, which are involved in glycolysis and hydrogen peroxide production, were identified in the culture supernatant of the Δply mutant. Neither the wild type nor the mutant strain could fragment the mycelium in the presence of catalase. These data suggest that hydrogen peroxide could fragment the mycelium and would terminate the co-existence of A. fumigatus and S. pneumoniae in biofilm.

Lactoperoxidase-generated hypothiocyanite efficiently kills Streptococcus pneumoniae

Abstract Streptococcus pneumoniae (pneumococcus) infections affect millions of people worldwide, cause serious mortality and represent a major economic burden. Airway epithelial cells, the primary responders to pneumococcal infection, orchestrate an extracellular antimicrobial system consisting of lactoperoxidase, thiocyanate anion and hydrogen peroxide (H2O2). Lactoperoxidase oxidizes thiocyanate using H2O2 into the final product hypothiocyanite. While hypothiocyanite has known antimicrobial effects against a wide range of microorganisms, its effect on pneumococcus has never been studied. To test this, pneumococci were exposed to hypothiocyanite generated enzymatically in a cell-free in vitro system. Bacterial killing was determined by colony forming units on agar plates while bacteriostatic effects were measured by changes in optical density of liquid cultures. We demonstrate hypothiocyanite exerted robust killing of several pneumococcal strains. Nonencapsulated pneumococcal mutants were killed to the same extent as their parental strains, indicating no role of the capsule in protection against hypothiocyanite. Catalase, an H2O2 scavenger, inhibited pneumococcal killing by hypothiocyanite under all circumstances. Interestingly, a pneumococcal mutant deficient in pyruvate oxidase, the main bacterial H2O2 source, had enhanced susceptibility to hypothiocyanite indicating the role of this gene in partial protection of the bacterium against this oxidizing agent. Overall, numerous pneumococcal strains were found to be susceptible to lactoperoxidase-generated hypothiocyanite in vitro, which presents an opportunity to explore hypothiocyanite as a potential novel anti-pneumococcal therapy.

Lung resident memory B cells are a common and functionally significant component of lung adaptive immunity

Abstract Resident memory B cells (BRM) in influenza-recovered mouse lungs were recently described, but whether other types of infections elicit these cells is unknown. The relevance of BRM in human lungs and to lung immune defenses also remains unexplored. Using flow cytometry and immunofluorescence, we found that respiratory pneumococcal exposures in mice elicited lung BRM without concurrent tertiary lymphoid structure formation. Additionally, flow cytometry analysis of normal human lung tissue showed that human lungs are enriched compared to human blood for B cells bearing a resident memory phenotype. These findings indicate that lung BRM are a common feature of antigen-experienced lungs. Multiple mouse models were used to address the contributions of B cell immunity to anti-pneumococcal lung defenses. Mice exposed to a low virulence pneumococcal strain 4 weeks previously were well-protected from a serotype-mismatched pneumococcal challenge. When previously exposed mice were depleted of circulating B cells (but not lung B cells) with anti-CD20 treatment before the challenge infection, there was no effect on the acquired lung immunity. However, a genetically engineered mouse strain allowed effective depletion of lung B cells bearing PD-L2 (a mouse memory B cell marker) from previously exposed mice, and doing so before the virulent pneumococcal challenge resulted in substantial defects in bacterial clearance compared to mice with lung B cells intact. These results provide the first direct evidence of a role for lung BRM in anti-bacterial lung immunity. Notably, this defense was pneumococcal serotype-independent, distinguishing it from the serotype-specific immunity elicited by current pneumococcal vaccines.

The impact of PCV7/13 on the distribution of carried pneumococcal serotypes and on pilus prevalence; 14 years of repeated cross-sectional surveillance

BackgroundS. pneumoniae carriage by children is a major source of pneumococcal transmission, and the initial step prior to infection. Pilus type 1, reported in ~30% of pneumococcal strains in the pre-vaccine era, contributes to pneumococcal colonization and virulence. In this study, we report the impact of the pneumococcal conjugate vaccine (PCV), PCV7/PCV13 sequential implementation on serotype distribution, and on the prevalence of piliated strains among carried pneumococci during the pre- and post-vaccine eras. MethodsDuring 2002-2016, 12 repeated cross-sectional surveillances of nasopharyngeal S. pneumoniae carriage were conducted among 8,473 children <5.5 years old visiting primary care physicians in Central Israel. Seven biannual surveillances in the pre-PCV period, 2 surveillances after PCV7 was licensed but before implementation in the National Immunization Plan, and 3 additional surveillances in the post-PCV period. S. pneumoniae serotype distribution and prevalence of piliated strains were assessed. ResultsCarriage of S. pneumoniae was relatively stable (45.4%). The prevalence of serotypes included in PCV13 was 65.7%, in the pre-vaccine period and the pilus was present in 26.4% of isolates. The distribution of serotypes and the pilus prevalence in the pre-PCV period was relatively stable except for a decrease in prevalence of piliated 19F, observed following the first study year. Following PCV7/PCV13 implementation, vaccine type 13 (VT13) strains were nearly eliminated to 3.3% by 2016. Piliated strains, which were primarily of VT13 serotypes, initially followed a similar trend and were nearly eliminated by 2014 (1.7%). Yet, two years later, pilus prevalence re-emerged among non-VT strains to 12.8% of all pneumococci. ConclusionsFollowing PCV implementation, a dramatic and rapid decrease in VT strains prevalence was observed with a concomitant increase in non-VT strains. Piliated strains were nearly eliminated, yet re-emerged 7 years following PCV7/PCV13 implementation in various non-VT strains. This suggests that the pilus confers an advantage in colonization.

Evolución de la distribución de serotipos de Streptococcus pneumoniae aislados en líquido pleural en la Comunidad de Madrid entre los años 2007 y 2018

IntroductionThe aim of this study was to describe the distribution of Streptococcus pneumoniae serotypes in isolates from pleural fluid in the Madrid Autonomous Community between the years 2007-2018. MethodsInvasive pneumococcal disease strains isolated during the period 2007-2018 were studied. The frequency of serotypes from pleural fluid was compared with that observed in other samples. ResultsA total of 6,115 pneumococcal invasive isolates were processed. Of them, 182 (3%) were isolated from pleural fluid. A total of 70.9% of isolates belonged to some of the following 6serotypes: 1, 3, 19A, 8, 7F and 5. The serotypes 3 and 8 increased significantly from 9.6% to 30.8%, and from 5.3% to 20.5%, respectively, over the periods 2007-2010 to 2015-2018. ConclusionsPneumococcal serotypes 3 and 8 are currently significant causes of infection of pleural fluid in our region.

Virulent Properties of Serotypes of Streptococcus pneumoniae from Child Carriers in the Republic of Tatarstan

The paper covers research results evaluating the occurrence of various serotypes of circulating pneumococcal strains in preschool children in the Republic of Tatarstan in 2017–2018, the dominance of “vaccine” serotypes and the emergence of new “non-vaccine” strains are shown. It has been shown that 86.5% of clinical isolates of Streptococcus pneumoniae from healthy carrier children have pronounced IgA-protease activity. Serotypes 14, 19F, 7F, 23F, 16F have high protease activity. Discrepancies in protein expression between strains with different levels of IgA protease activity were studied using differential gel electrophoresis followed by mass spectrometric identification. Study of the protein expression profiles by gel electrophoresis in S. pneumoniae strains with different proteolytic activity resulted in the identification of 5 protein spots which showed a significant difference in expression (at least a 2-fold change between the two phenotypes).

RETRACTED: Co infection of respiratory syncytial viruses (RSV) and Streptococcus pneumonia modulates pathogenesis and dependent of serotype and phase variant

Streptococcus pneumoniae (pneumococcus) is touted to be the generally found pathogen in patients with respiratory issues and there is an epidemiologic linkage present between Respiratory syncytial virus (RSV). This study aim at investigating the interaction between RSV and two serotypes of S. pneumoniae using a distinct animal model and a well-established colonizing pneumococcal strain. Phase variants phenotype of each strain was determined under oblique light. Co infection model was developed using BALB/c mice housed in a BSL-2 facility. Coinfection experiments were performed and number of bacterial colonies was quantified and phase determination was evaluated. RSV was detected in sample through real-time quantitative PCR. Adherence assays were performed to determine adherence of Spn strains and its knock out ΔNanA to nasopharyngeal carcinoma (NPC) epithelial CNE3 cell line. The biofilm viability was determined and phase composition was counted using plate count. Neuraminidase activity was measured in fluorometircassessed using 2'-(4-methylumbelliferyl)-α-D-N-acetylneuraminic acid (MUAN) as substrate as described in earlier literature. The GraphPad Software version 5.01 i.e., GraphPad Prism was used to conduct the statistical analysis. The extent of bacterial colonization was increased significantly (p<0.05), when the mice were co infected. Nasal epithelium remained intact in mock sample with features of a thick mucociliary border. A small percentage of pneumococci exhibit phase variation between opaque phase and transparent phase. The percentage adherent of both phase were not found to be varying significantly within serotype but it was seen that nonpathogenic type 27 was more adherent. Biofilm formation was selectively more for transparent phase from a mixed-phase inoculum. Adherence of both phase variant of S. pneumoniae to nasopharyngeal epithelial cells 2h post infection expressed as the percentage of adherent bacteria relative to the inoculum. In absence of viral infection, the nasal colonization of the opaque and the transparent variant was increased many folds, which was a significant differences. The extent of nasal colonization by the ΔNanA mutant strain were significantly reduced post-bacterial infection for both type of wild-type (P<0.05). The findings explore insights into the interactions occurring between S. pneumoniae and RSV during respiratory infections and pneumococcal acquisition, indicate that pneumococcal serotypes have different ability to cause infection as well as co infections and potentially follow an unappreciated mechanism. Much more research work is needed to further understand the minutiae of this interaction within co-infection process.

Process intensification for production of Streptococcus pneumoniae whole-cell vaccine.

The available pneumococcal conjugate vaccines provide protection against only those serotypes that are included in the vaccine, which leads to a selective pressure and serotype replacement in the population. An alternative low-cost, safe and serotype-independent vaccine was developed based on a nonencapsulated pneumococcus strain. This study evaluates process intensification to improve biomass production and shows for the first time the use of perfusion-batch with cell recycling for bacterial vaccine production. Batch, fed-batch, and perfusion-batch were performed at 10 L scale using a complex animal component-free culture medium. Cells were harvested at the highest optical density, concentrated and washed using microfiltration or centrifugation to compare cell separation methods. Higherbiomass was achieved using perfusion-batch, which removes lactate while retaining cells. The biomass produced in perfusion-batch would represent at least a fourfold greater number of doses per cultivation than in the previously described batch process. Each strategy yielded similar vaccines in terms of quality as evaluated by western blot and animal immunization assays, indicating thatso far, perfusion-batch is the best strategy for the intensification of pneumococcal whole-cell vaccine production, as it can be integrated to the cell separation process keeping the same vaccine quality.

Evaluation of protective immunity responses against pneumococcal PhtD and its C-terminal in combination with outer-membrane vesicles as adjuvants.

Introduction. Streptococcus pneumoniae is a significant bacterial pathogen in humans. Currently, there are two types of pneumococcal vaccines, but there are concerns regarding their application.Aim. Since many pneumococcal proteins are serotype-independent, polyhistidine triad protein D (PhtD) has been selected as a vaccine candidate.Methodology. We prepared recombinant PhtD and its C-terminal fragment (PhtD-C) using alum and outer-membrane vesicles (OMVs) as adjuvants. The combinations were injected intraperitoneally into mice, and then total immunoglobulin G (IgG) and specific IgG, IgG1 and IgG2a were measured. A serum bactericidal assay and opsonophagocytosis were also performed as complementary tests. Meningococcal OMVs were used as an adjuvant.Results. The levels of specific IgG and IgG1 against combinations of PhtD and its C-terminal with OMVs and alum as adjuvants increased at the time of the third mouse immunization on day 35. Forty per cent and 60% of S. pneumoniae ATCC 6303 (serotype 3) as a virulent pneumococcal strain, respectively, were killed in the opsonophagocytosis test and these results could also be observed in the serum bactericidal assay. Mice mmunized iwith PhtD and its C-terminal with OMVs and alum as adjuvants survived after 10 days of pneumococcal challenge.Conclusion. The combination of PhtD and PhtD-C with alum produced optimal results, but the combination of PhtD and PhtD-C with OMVs produced minimal results by comparison. The survival rates were also measured, and these corresponded with the results of the immunological assessments. Our findings showed that mice receiving PhtD and PhtD-C plus OMV and alum had higher survival rates than the mice in the other groups.

Rising Pneumococcal Antibiotic Resistance in the Post-13-Valent Pneumococcal Conjugate Vaccine Era in Pediatric Isolates From a Primary Care Setting.

Antibiotic-resistant Streptococcus pneumoniae strains may cause infections that fail to respond to antimicrobial therapy. Results reported from hospitalized patients with invasive, bacteremic infections may not be the same as those observed in a primary care setting where young children receive care for noninvasive infections. Young children experience the highest burden of pneumococcal disease. The aim of this study was to determine the antibiotic susceptibility of S. pneumoniae strains isolated from children in a primary care setting in the post-13-valent pneumococcal conjugate vaccine (PCV13) era. This was a prospective collection of 1201 isolates of S. pneumoniae from 2006 through 2016 in a primary care setting. Antibiotic susceptibility testing to 16 different antibiotics of 10 classes was performed. Participants were children aged 6-36 months. Nasopharyngeal swabs were obtained from patients during acute otitis media (AOM) visits and routine healthy visits. Middle ear fluid was obtained by tympanocentesis. After introduction of PCV13, antibiotic susceptibility of pneumococci, especially to penicillin, initially improved largely due to disappearance of serotype 19A, included in PCV13. However, beginning in 2013, antibiotic susceptibility among pneumococcal strains began decreasing due to new serotypes not included in PCV13. In addition to reduced susceptibility to penicillin, the most recent isolates show reduced susceptibility to third-generation cephalosporins, fluoroquinolones, and carbapenems, antibiotics commonly used to treat life-threatening, invasive pneumococcal diseases. In recent years, pneumococcal nasopharyngeal and AOM isolates from children exhibit reduced susceptibility to penicillin, third-generation cephalosporin, fluoroquinolone, and carbapenem antibiotics. The new strains have a different profile of resistance compared to the pre-PCV13 era.

How nasopharyngeal pneumococcal carriage evolved during and after a PCV13-to-PCV10 vaccination programme switch in Belgium, 2016 to 2018.

BackgroundThe current carriage study was set up to reinforce surveillance during/after the PCV13-to-PCVC10 switch in Belgium.AimThis observational study monitored carriage of Streptococcus pneumoniae (Sp) serotypes, particularly those no longer covered (3, 6A, 19A), as well as Haemophilus influenzae (Hi), because PCV10 contains the non-typeable Hi protein D.MethodsA total of 2,615 nasopharyngeal swabs from children (6–30 months old) attending day care were collected in three periods over 2016–2018. Children’s demographic and clinical characteristics and vaccination status were obtained through a questionnaire. Sp and Hi were identified by culture and PCR. Pneumococcal strains were tested for antimicrobial (non-)susceptibility by disc diffusion and serotyped by Quellung-reaction (Quellung-reaction and PCR for serotypes 3, 6A, 19A).ResultsThe carriage prevalence of Sp (> 75%) remained stable over the successive periods but that of Hi increased (87.4%, 664 Hi-carriers/760 in 2016 vs 93.9%, 895/953 in 2017–2018). The proportion of non-PCV13 vaccine serotypes decreased (94.6%, 438 isolates/463 in 2016 vs 89.7%, 599/668 in 2017–2018) while that of PCV13-non-PCV10 vaccine serotypes (3 + 6A + 19A) increased (0.9%, 4 isolates/463 in 2016 vs 7.8%, 52/668 in 2017–2018), with serotype 19A most frequently identified (87.9%, 58/66 isolates). Non-susceptibility of pneumococci against any of the tested antibiotics was stable over the study period (> 44%).ConclusionsDuring and after the PCV13-to-PCV10 vaccine switch, the proportion of non-PCV13 serotypes decreased, mainly due to a serotype 19A carriage prevalence increase. These results complement invasive pneumococcal disease surveillance data, providing further basis for pneumococcal vaccination programme policy making.

Serotype and clonal distribution dynamics of invasive pneumococcal strains after PCV13 introduction (2011-2016): Surveillance data from 23 sites in Catalonia, Spain.

BackgroundThe objective of this study is to describe incidence and shifts of serotype and clonal distribution of invasive Streptococcus pneumoniae strains in four different age groups (<5 years, 5–17 years, 18–64 years and >65 years) during a period of intermediate PCV13 vaccination coverage (2011–2016) in Catalonia, Spain.MethodsWe included all pneumococcal strains systematically sent to the Catalan support laboratory for molecular surveillance of invasive pneumococcal disease (IPD) located at Hospital Sant Joan de Deu, Barcelona. Two study periods were considered: 2011–13, early PCV13 vaccination period (EVP) and 2014–2016, late vaccination period (LVP).ResultsA total of 2142 strains were included in the study. Five years after intermediate introduction of PCV13 in our population, a significant decrease of overall incidence of IPD in children <5 years was observed (incidence rate ratio 0.5, 95% confidence interval 0.4–0.8). However, in seniors older than 65 years, a significant increase of overall incidence of IPD was observed (IRR 1.4, 95% CI 1.1–1.7). The contribution of PCV13 vaccine serotypes to IPD declined significantly in all age groups: from 59% to 38.1% in <5 years; 82.7% to 59% in 5–17 years, 47.8% to 34.1% in 18–64 years and 48.2% to 37% in >65 years. Results found when comparing both periods were consistent with IRRs observed year by year. In children <5 years, the three major serotypes detected were 1, 24F and 19A in EVP vs 24F, 14 and 10A in LVP. Among patients 5–17 years the first three serotypes were 1, 12F and 14 both in EVP and LVP. Among adults 18–64, the three major serotypes detected were 1, 12F and 8 vs 8, 12F and 3, respectively. Finally, in patients >65 years the most frequently isolated serotypes were 3, 19A and 7F vs 3, 14 and 12F, respectively. Regarding clonal complexes (CCs) expressing mainly PCV13 serotypes, significant decreases of the proportions of CC306, CC191 and CC320 were observed, while CC156 showed a significant increase. As for CCs expressing mostly non-PCV13 serotypes, significant increases in ST989, CC53 and CC404 were showed.ConclusionsDespite low vaccine coverage in our setting a significant decrease of incidence of IPD was observed in children younger than 5 years. The modest indirect protection against vaccine serotypes causing IPD in elderly indicate the need for the inclusion of more serotypes in future high-valent PCV and vaccinating old adults should be considered.

Impact of thirteen-valent pneumococcal conjugate vaccine on nasopharyngeal carriage in healthy children under 24 months in Okinawa, Japan

In November 2013, a 13-valent pneumococcal conjugate vaccine (PCV13) for all infants aged younger than 5 years was incorporated into the Japan national immunization program. An annual survey of nasopharyngeal carriage rates was performed on healthy infants aged 2–24 months from Okinawa, Japan to evaluate the effect of PCV13 on pneumococcal colonization. Of 756 evaluable infants, 203 pneumococcal strains were detected in 193 infants. The overall nasopharyngeal carriage rate was 25.5%, which was not different from our previously reported isolation rate before the introduction of PCV13. The main serotypes of the Streptococcus pneumoniae strains are 15A (18.2%), non-typeable (14.8%), and 15B (11.8%). The carriage rates of pneumococcal strains with 7-valent pneumococcal conjugate vaccine serotypes and PCV13 serotypes were 3.0% and 9.9%, respectively. These values were significantly lower than we reported before the introduction of PCV13. However, the carriage rates of non-PCV13 serotypes have increased. Multivariate logistic regression analysis suggested that siblings and day care attendance are risk factors for pneumococcal carriage.

Chimeric oligosaccharide conjugate induces opsonic antibodies against Streptococcus pneumoniae serotypes 19A and 19F.

Streptococcus pneumoniae 19A (ST19A) and 19F (ST19F) are among the prevalent serotypes causing pneumococcal disease worldwide even after introduction of a 13-valent pneumococcal conjugate vaccine (PCV13). Synthetic glycoconjugate vaccines have defined chemical structures rather than isolated polysaccharide mixtures utilized in marketed vaccines. Ideally, a minimal number of synthetic antigens would cover as many bacterial serotypes to lower cost of goods and minimize the response to carrier proteins. To demonstrate that a chimeric oligosaccharide antigen can induce a protective immune response against multiple serotypes, we synthesized a chimeric antigen (ST19AF) that is comprised of a repeating unit of ST19A and ST19F capsular polysaccharide each. Synthetic glycan epitopes representing only ST19A, and ST19F were prepared for comparison. Semisynthetic glycoconjugates containing chimeric antigen ST19AF induced high antibody titers able to recognize native CPS from ST19A and ST19F in rabbits. The antibodies were able to kill both strains of pneumococci. Chimeric antigens are an attractive means to induce an immune response against multiple bacterial serotypes.

Факторы риска формирования резистентности Streptococcus pneumoniae к антибиотикам

Streptococcus pneumoniae is responsible for a wide range of human diseases, often with a severe course and prognosis. The growing resistance of pneumococci to certain classes of antibiotics limits the treatment of pneumococcal infections. The study of factors that influence the formation of antibiotic resistance of the pathogen is important for developing a strategy to counter this phenomenon and for predicting the presence of microbial resistance in a particular patient when choosing a drug for empirical therapy. This review presents the results of studies of some potential risk factors for the formation of Streptococcus pneumoniae resistance to antibiotics. The most important of these is the previous administration of antibiotics, especially in the last few months. Previous diseases, previous hospitalization, and the nosocomial nature of pneumonia are also considered as factors that allow predicting the colonization of loci by antibiotic-resistant strains of pneumococci. Staying at long-term care facilities and attending organized groups (schools and kindergartens) may also increase the risk of colonization of the respiratory tract by antibiotic-resistant strains of pneumococci. The frequency of isolation of pneumococcus strains insensitive to certain antibiotics is also characterized by wide geographical variability.

Karakterizacija sojeva Streptococcus pneumoniae izolovanih kod pacijenata sa meningitisom pre uvođenja pneumokokne konjugovane vakcine u Srbiji

Introduction: Pneumococcal meningitis is a serious disease which affects mostly children ≤ 2 years, adults ≥ 65 years and immunodeficient patients. The introduction of pneumococcal conjugate vaccine (PCV) into immunization programs worldwide has led to a significant decrease in the incidence of invasive pneumococcal disease, reduction of antibiotic resistance and changes in the distribution of pneumococcal serotypes. In 2018, PCV10 was introduced into the National Immunization Program in Serbia. Aim: The aim of this study was to analyze the serotype distribution and antibiotic susceptibility of pneumococcal strains isolated from meningitis cases in the pre-vaccinal period (2009-2018) in Serbia. Material and methods: Meningeal isolates were sent into the National Reference Laboratory for streptococci between January 2009 and December 2018 for serotyping and antimicrobial susceptibility testing (AST). Serotyping was performed by Quellung reaction, while AST was performed using disk diffusion method and E-test. Results: A total of 199 strains were analyzed and 32 different serotypes have been identified. Among the 55 (27.64%) strains from children ≤16 years, 17 different serotypes were detected of which 19F, 14, 6A, and 6B were the most common. Regarding the 144 (72.36%) adult isolates, 30 different serotypes were present, the most common being 3, 19F, 14, 23F, 6A and 6B. The coverage of pediatric serotypes was 61.82% for PCV10 and 78.18% for PCV13. Statistically significant number of isolates showed resistance to: penicillin (53.26%), erythromycin (45.73%), clindamycin (40.20%), trimethoprim-sulfamethoxazole (34.17%) and tetracycline (34.17%). The isolates from children were more resistant to beta-lactams and macrolides (p &lt; 0.05). Conclusion: During the pre-vaccinal period in Serbia, vaccinal serotypes dominated over non-vaccinal serotypes. Resistance is intermediate to high in the dominant serotypes of children and low in the most common adult serotype 3. Taking into consideration the possible changes in the pneumococcal population in the future, continued monitoring of post-vaccine serotype and resistance trends are essential.

Invasive pneumococcal disease among hospitalized children in Brazil before and after the introduction of a pneumococcal conjugate vaccine

BackgroundMost of the available data on invasive pneumococcal disease in Latin America are derived from laboratory-based surveillance systems. There is a lack of epidemiological data on the disease severity and mortality from hospitalized patients with pneumococcal infection. MethodsIn this hospital-based retrospective historical series of hospitalized children with laboratory-confirmed IPD, we evaluated changes in disease episodes, in-hospital fatality rates, and need for intensive care unit admission after the inclusion of PCV10 in the Brazilian vaccination schedule. Invasive pneumococcal strains isolated by culture were serotyped. Changes over time were assessed, and pre-vaccination (2005–2009) to post-vaccination (2011–2015) disease rates and serotypes were compared. Results260 patients with IPD and positive pneumococcal isolates were identified (198 during the pre-PCV10 period). When comparing both periods, hospitalizations were reduced from 20 cases to 5 cases per 10,000 pediatric admissions (p < 0.0001). Likewise, fatalities reduced from 6.6 to 2.0 cases per 10,000 pediatric admissions (p < 0.0001). Pneumonia was the most frequent clinical diagnosis (58%) – of which 49.6% had pleural effusion – followed by meningitis (22%) and bacteremia (15.9%). Overall 30% of cases were sent to ICU, with no percentual changes after PCV10. Additional PCV13 serotypes increased from 7% before vaccine introduction to 21% after PCV10 use. Similarly, serotypes not included in PCV13 increased from 11% to 29%. ConclusionsThere was a significant reduction in the hospitalizations rates, ICU admissions, and fatalities due to IPD after PCV10 introduction in Brazil. Cases due to PCV10 serotypes were reduced, while infections rates caused by non-PCV10 serotypes increased.