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BACKGROUNDNasopharyngeal carriage of pathogenic bacteria precedes invasive disease and higher rates are found in low socioeconomic-status (SES) settings. Local immune responses are important for controlling colonization, but whether SES affects these responses is currently unknown.OBJECTIVEExamining bacterial colonization and cytokine response in nasal mucosa of children from high and low SES.METHODSTwenty-five cytokines were measured in nasal fluid. qPCR was performed to determine carriage and density of Haemophilus influenzae (H. influenzae), Streptococcus pneumoniae (S. pneumoniae), Moraxella catarrhalis (M. catarrhalis) and Staphylococcus aureus (S. aureus).RESULTSThe densities of H. influenzae and S. pneumoniae were increased in low compared to the high SES (p=0.006, p=0.026), with respectively 6 and 67 times higher median densities. Densities of H. influenzae and S. pneumoniae were positively associated with levels of IL-1beta (p=0.002, p=0.008) and IL-6 (pp=0.006). After correcting for bacterial density, IL-6 levels were increased in colonized children from high compared to low SES for both H. influenzae and S. pneumoniae (both p=0.039).CONCLUSIONIncreased density of H. influenzae and S. pneumoniae was observed in low SES children, while IL-6 levels associated with colonization were reduced in these children, indicating that immune responses to bacterial colonization were altered by SES.

Aims To explore the in vitro ability of alpha haemolytic streptococcus (AHS) and lactobacilli (LBs), from Indigenous Australian children, to inhibit the growth of respiratory pathogens (Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis), also from Indigenous Australian children. Methods and Results The bacterial interference of 91 isolates, from Indigenous Australian children both with and without otitis media (OM) or rhinorrhoea, was investigated using agar overlay and cell-free supernatant. Promising isolates underwent whole genome sequencing to investigate upper respiratory tract tropism, antibiotic resistance and virulence. Antibiotic susceptibility was examined for ampicillin, amoxicillin +clavulanic acid and azithromycin. Differences in the strength of bacterial inferences in relation to OM was examined using a case series of three healthy and three children with OM. LBs readily inhibited the growth of pathogens. AHS were less effective, although several isolates inhibited S. pneumoniae. One L. rhamnosus had genes coding for pili to adhere to epithelial cells. We detected antibiotic resistance genes coding for antibiotic efflux pump and ribosomal protection protein. LBs were susceptible to antimicrobials in vitro. Screening for virulence detected genes encoding for two putative capsule proteins. Healthy children had AHS and LB that were more potent inhibitors of respiratory pathogens in vitro than children with OM. Conclusions L. rhamnosus from remote Indigenous Australian children are potent inhibitors of respiratory pathogens in vitro. Significance and Impact of Study Respiratory/ear disease are endemic in Indigenous Australians. There is an urgent call for more effective treatment/prevention; beneficial microbes have not been explored. L. rhamnosus investigated in this study are potent inhibitors of respiratory pathogens in vitro and require further investigation.

Objectives: Omadacycline was tested against 7000 bacterial isolates collected prospectively from medical centres in the USA during 2019. Methods: Antimicrobial susceptibility testing was performed according to Clinical and Laboratory Standards Institute (CLSI) guidelines. Results: Omadacycline was active against: Staphylococcus aureus (MIC50/90, 0.12/0.25 mg/L; 98.3% susceptible), including methicillin-resistant S. aureus (MRSA); Enterococcus faecalis (MIC50/90, 0.06/0.25 mg/L; 100.0% susceptible), including vancomycin-resistant enterococci (VRE); Streptococcus pneumoniae (MIC50/90, 0.06/0.06 mg/L; 99.8% susceptible); viridans group streptococci, including Streptococcus anginosus group (MIC50/90, 0.03/0.06 mg/L; 100.0% susceptible); β-haemolytic streptococci, including Streptococcus pyogenes (MIC50/90, 0.06/0.12 mg/L; 99.2% susceptible); Enterobacterales (MIC50/90, 1/8 mg/L; 86.9% inhibited at ≤4 mg/L), including Escherichia coli (MIC50/90, 0.5/2 mg/L; 99.6% inhibited at ≤4 mg/L); Enterobacter cloacae (MIC50/90, 2/4 mg/L; 98.5% susceptible); Klebsiella pneumoniae (MIC50/90, 1/4 mg/L; 93.2% susceptible); Acinetobacter baumannii (MIC50/90, 0.5/4 mg/L; 90.8% inhibited at ≤4 mg/L); Haemophilus influenzae (MIC50/90, 0.5/1 mg/L; 100.0% susceptible); and Moraxella catarrhalis (MIC50/90, ≤0.12/0.25 mg/L). Conclusion: The 2019 in vitro activity of omadacycline against key Gram-positive and Gram-negative pathogens has not changed compared with the prior 3 years of surveillance in the SENTRY Antimicrobial Surveillance Program. Omadacycline merits further study in serious infections where resistant pathogens may be encountered.

Study objective To study the bacteria and fungi causing vulvovaginitis in prepubertal girls. Design Swabs from vaginal introitus were collected from patients with vulvovaginitis in 2018, and cultured for the identification of microorganisms with standard microbiological techniques. Setting A children's hospital in Hangzhou, East China. Participants A total of 1235 Chinese prepubertal girls diagnosed with vulvovaginitis. Main Outcome Measure Bacteria or fungi in pure cultures or as the predominant organism were defined as pathogens. Results A total of 1235 cases were diagnosed as vulvovaginitis, and 515 isolates were identified from 494 patients (40%, 494/1235). The most common pathogen was Streptococcus pyogenes (27.6%,142/515), followed by Haemophilus influenzae (27.2%,140/515), Candida albicans (22.3%,115/515), Staphylococcus aureus (5.8%, 30/515) and Escherichia coli (4.7%,24/515). All S pyogenes isolates were sensitive to penicillin, whereas 53.7% (73/136) of H influenzae isolates were sensitive to ampicillin, and 70.4% (19/27) of S aureus isolates were sensitive to oxacillin. Conclusion S pyogenes and H influenzae were the 2 most commonly identified pathogenic bacteria found in prepubertal girls with vulvovaginitis. Vulvovaginitis in prepubertal girls caused by C albicans may be more common in older children.

On the first of January 2019, the European Committee on Antimicrobial Susceptibility Testing, EUCAST, introduced the concept of "area of technical uncertainty" (ATU). The aim was to report on the incidence of ATU test results in a selection of common bacterial species and the subsequent impact on antimicrobial resistance categorization and workload. A retrospective analysis of clinical samples collected from February 2019 until November 2019 was performed. Susceptibility to amoxicillin-clavulanic acid and piperacillin-tazobactam in Enterobacterales (Escherichia spp., Klebsiella spp., Proteus spp.), piperacillin-tazobactam in Pseudomonas aeruginosa, and amoxicillin-clavulanic acid and cefuroxime in Haemophilus influenzae was studied. Disk diffusion antibiotic susceptibility testing was read and interpreted by ADAGIO 93400 automated system (Bio-Rad, France). In case of an inhibition zone in the ATU, strains were retested using gradient minimal inhibitory concentration method (Etest, BioMerieux, France). Overall, 14,164 isolate-antibiotic combinations were tested in 7922 isolates, resulting in 1204 (8.5%) disk zone diameters in the ATU region. Retesting of ATUs with Etest resulted in a category change from S to R for amoxicillin-clavulanic acid in 63/498 (12.7%) of Escherichia spp., 2/58 (3.4%) of Klebsiella spp., 2/37 (5.4%) of Proteus spp., and 6/125 (4.8%) of Haemophilus influenzae. For piperacillin-tazobactam, a category change from S to R was found in 33/92 (35.9%) of Pseudomonas aeruginosa. We conclude that ATU testing has a substantial impact on the correct interpretation of antimicrobial resistance, at the expense of turn-around time and with the cost of additional workload.

Bacterial co-infections increase the severity of respiratory viral infections and were frequent causes of mortality in influenza pandemics, but have not been well characterized in patients with coronavirus disease 2019 (COVID-19). The aim of this review was to identify the frequency and microbial etiologies of bacterial co-infections that are present upon admission to the hospital and that occur during hospitalization for COVID-19. We found that bacterial co-infections were present in

Background The role of nasopharyngeal bacteria in respiratory syncytial virus (RSV) disease has been underestimated. We measured the frequency and burden of respiratory bacteria in the upper respiratory tract of infants with RSV infection over 7 respiratory seasons, and their impact on clinical outcomes. Methods Children Results S. pneumoniae, H. influenzae, and M. catarrhalis, but not S. aureus, were detected more frequently in RSV-infected children (84%) than healthy controls (46%; P Conclusions Nasopharyngeal codetection of S. pneumoniae and H. influenzae in infants with RSV infection is associated with increased disease severity.

Background. Upper respiratory tract infections (URTIs) are one of the most common diseases of childhood. It is known that some children tend to have recurrent bacterial complications of URTIs; the liability mentioned might be linked to different factors including local mucosal immune resistance malfunction. Infectious respiratory pathology is currently being considered comprehensively in the aspect of viral-bacterial-immune interaction, consequently the study of microbiological and immunological factor relations may be helpful in promoting the development of targeted therapeutic and preventive measures that may improve the efficiency of the management of such patients. Thereby, the study objectives were to investigate the local mucosal resistance state, features of microbial colonization of upper airways along with etiological structure complications in children susceptible to repeated bacterial complications of URTIs. Materials and methods. We enrolled 214 children aged 3 to 18 years with URTIs complicated with acute otitis media (AOM) or acute bacterial rhinosinusitis (ARS). Rate of bacterial complications of URTIs in 128 children was low (group I) and in 86 children it met the criteria of recurrent course (group II). We performed microbiological examination of nasopharyngeal secretions or middle ear exudates as well as local mucosal immunology study: lysozyme, lactoferrin, human cathelicidin hCap-18/LL-37, secretory IgA concentrations in oropharyngeal discharge measured twice during the disease, and after recovery. The controls were 36 children with uncomplicated URTIs, and the reference values were obtained in 30 healthy children. Results. In children with recurrent (versus episodic) bacterial complications of URTIs malfunction of airway local protective innate mucosal immune mechanisms were present: reduction of human cathelicidin level during the whole period of disease, reduction of lactoferrin at the early stages of bacterial process, and reduced concentrations of lysozyme at all phases of the study. It’s noteworthy that concentrations of hCAP/LL-37 and lactoferrin during the disease exceeded control and referent values in both groups; while lysozyme levels obtained from children of the II group were significantly lower compared to referent patients both initially and after recovery. The levels of antimicrobial peptides demonstrated the inverse correlation with URTI symptoms duration. In the II group the level of negative microbiological results for causative pathogen of AOM/ARS was about 35 % vs. about 20 % in group I (p = 0.008). Also lower rate of Streptococcus pneumoniae and nontypeable Haemophilus influenzae (pathogenic bacteria), and higher rate of Haemophilus parainfluenzae and Staphylococcus aureus (associated agents) isolates occurred in group II. Furthermore, associations of two non-causative bacterial agents were obtained only in group II. Non-causative bacteria colonization in general was associated with lower levels of mucosal antimicrobial peptides, but on the background of URTI it might activate tempora­rily lysozyme and lactoferrin response. Conclusions. Recurrent bacterial complications of URTIs are associated with local mucosal resistance insufficiency, and higher rate of St.aureus and H.parainfluenzae colonization. High rate of microbiological failures in determining of causative agent of AOM/ARS may be related to over diagnostics of bacterial complications which might be linked to misinterpretation of prolonged course of the disease due to mucosal immune dysfunction and non-causative bacteria colonization. Thus, di­sease duration shouldn’t be used as an independent criterion of bacterial process in this category of patients because it may lead to misdiagnose and antibiotic overuse.

Haemophilus influenzae is a leading cause of meningitis disease and mortality, particularly in young children. Since the introduction of a licensed conjugate vaccine (targeting the outer capsular polysaccharide) against the most prevalent serotype, Haemophilus influenzae serotype b, the epidemiology of the disease has changed and Haemophilus influenzae serotype a is on the rise, especially in Indigenous North American populations. Here we apply molecular modeling to explore the preferred conformations of the serotype a and b capsular polysaccharides as well as a modified hydrolysis resistant serotype b polysaccharide. Although both serotype b and the modified serotype b have similar random coil behavior, our simulations reveal some differences in the polysaccharide conformations and surfaces which may impact antibody cross-reactivity between these two antigens. Importantly, we find significant conformational differences between the serotype a and b polysaccharides, indicating a potential lack of cross-reactivity that is corroborated by immunological data showing little recognition or killing between heterologous serotypes. These findings support the current development of a serotype a conjugate vaccine.

Background/Purpose Ceftaroline, with a unique activity against methicillin-resistant Staphylococcus aureus (MRSA), was not launched in Taiwan before 2019. The in vitro susceptibility data of ceftaroline against important Taiwanese pathogens are lacking. Methods The in vitro susceptibility of ceftaroline against important pathogens collected from 2012 through 2018 were extracted from the Antimicrobial Testing Leadership and Surveillance program. Broth microdilution method was used to determine the minimum inhibitory concentrations (MICs) to ceftaroline against all isolates. Results During the study period, the in vitro data regarding isolates of S. aureus (n = 2049), Staphylococcus epidermidis (n = 185), Streptococcus pneumoniae (n = 334), Streptococcus pyogenes (n = 170), Haemophilus influenzae (n = 75), Haemophilus parainfluenzae (n = 10) and Klebsiella pneumoniae (n = 680) regardless of hospital sites of collection were analyzed. Among the S. aureus isolates studied, 19.4% showed MICs of 1 mg/L to ceftaroline, and 4.4% showed in vitro susceptible-dose dependent to ceftaroline (all MICs, 2 mg/L). Most of other Gram-positive cocci, all H. influenzae and H. parainfluenzae isolates were susceptible to ceftaroline. By contrast, about one-third (35.9%) of K. pneumoniae isolates, irrespective of infection sources, exhibited non-susceptibility to ceftaroline (MIC range, 0.015–256 mg/L; MIC50 and MIC90 values, 0.12 and 256 mg/L, respectively). Conclusions From the pharmacodynamic perspectives, the ceftaroline dosage of 600 mg as a 2-h intravenous infusion every 8 h is effective against all S. aureus and other Gram-positive isolates regardless of acquisition sites in Taiwan. Before ceftaroline is prescribed in treatment of the patient with Gram-negative infection, a cautious evaluation about patient's healthcare-associated factor is warranted.

Objectives To identify microbiological growth on bicanalicular silicone tubes (BST) placed during dacryocystorhinostomy (DCR) surgery and to analyze the association between culture results and surgical outcomes and BST removal time. Materials and methods A total of 80 lacrimal drainage systems of 68 patients who had external DCR with bicanalicular silicone intubation were included the study. Twenty-five tubes (31.3%) were removed up to 8 weeks, 28 tubes (35.0%) were removed between 9 and 11 weeks, and the remaining 27 tubes (33.7%) were removed 12 weeks or more after surgery. The tubes were transferred to Stuart medium and sent for microbiologic examination. The disc diffusion method was used to determine antibiotic resistance. Results Culture positivity was observed for 96.2% of the tubes. Among a total of 109 isolates, 63 were gram-positive bacteria (57.8%), 37 were gram-negative bacteria (34%), and 9 were fungi (8.2%). The most commonly isolated gram-positive and gram-negative bacteria were Staphylococcus aureus (66.6%) and Enterobacter spp. (29.7%), respectively. Penicillin, clindamycin, erythromycin, and tetracycline resistances were higher among gram-positive pathogens. Cephalothin, amoxicillin-clavulanic acid, and ampicillin resistances were higher among gram-negative pathogens. There was no significant difference in terms of the microbiological profile between the three groups of removed tubes. Haemophilus influenzae was isolated at a significantly higher rate in patients with surgical failure (p=0.04). Conclusion Although a variety of agents were isolated from removed BST, gram-positive organisms were more frequent than gram-negatives and fungi. S. aureus and Enterobacter were the most common gram-positive and gram-negative isolates. Later BST removal was associated with the isolation of significantly more bacterial strains per tube. There was no correlation between multiple infections and surgical failure. H. influenzae was more common in failed DCR cases.

Background Bacterial culture is the gold standard for the diagnosis of invasive bacterial diseases (IBDs) but molecular methods are more specific and sensitive. Fresh liquid samples (FLSs) show patent limitations for shipping and storage. We aimed to evaluate the sensitivity and specificity of real-time polymerase chain reaction (PCR) performed on dried sample spots (DSSs) obtained from different biological fluids compared with real-time PCR or culture performed on FLSs. Methods FLSs positive for Streptococcus pneumoniae, Neisseria meningitidis, Haemophilus influenzae, Escherichia coli, Streptococcus pyogenes, Staphylococcus aureus, Bordetella pertussis and/or Pseudomonas aeruginosa were spotted on filter paper. Real-time PCR was performed on both FLSs and DSSs and results were compared. The stability of the DSS results over time was evaluated. Results Real-time PCR performed on 114 DSSs showed a specificity of 99.1% and a sensitivity of 91.2% for IBD diagnosis. A positive correlation was found between FLS cycle threshold (Ct) and DSS Ct (r=0.84; r2=0.71) with the Pearson statistical test and Bland–Altman analysis showing that 95% of the specimens were within agreeable limits. Although we observed a trend towards signal reduction over time in the DSSs, there was no statistical evidence of an increase in Ct values. Real-time PCR on DSSs was 2.2 times more sensitive than culture. Conclusions Real-time PCR applied to DSSs may be a useful approach in different situations, such as IBD diagnosis, both for rural areas of low-income countries and family practitioners in various settings.

Background Haemophilus influenzae serotype a (Hia) can cause severe invasive disease, especially in young children. In 2018, 4 invasive Hia cases occurred in an Alaska community. We used whole-genome sequencing (WGS) to evaluate the relationship of the bacteria from this community and other Alaska patients with invasive Hia. Methods All carriage (n = 15) and invasive (n = 4) Hia isolates from the outbreak community, together with 15 nonoutbreak Alaska invasive Hia surveillance isolates from 2018, were tested for antimicrobial susceptibility and characterized using WGS. Results Phylogenetic analysis of both invasive and carriage Hia isolates revealed 2 major clades that differed by an average of 300 core single-nucleotide polymorphisms (SNPs). All isolates from the outbreak community were clustered in 1 subclade, within a larger clade containing 3 nonoutbreak invasive Hia isolates. Comparative genomics did not reveal any genetic mutations that distinguished carriage from invasive isolates. Three (20%) community isolates were rifampin resistant and had a previously unreported mutation in the rpoB gene. Conclusions In the outbreak community, Hia isolates from carriers were indistinguishable from the invasive Hia isolates. Overall, invasive Hia isolates from Alaska in 2018 were genetically similar. The rifampin resistance mutation is concerning as rifampin is the first-line medication for Hia prophylaxis.

Lefamulin (Xenleta) has been approved by the US FDA for the treatment of community-acquired bacterial pneumonia (CABP). It may be taken intravenously or orally and has activity against a broad range of pulmonary pathogens, including Streptococcus pneumoniae, Haemophilus influenzae, Mycoplasma pneumoniae, Legionella pneumophila, and Chlamydophila pneumonia, as well as methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococcus faecium. Lefamulin has an adverse effect profile that is similar to other antimicrobial agents commonly used to treat CABP. Despite these promising features, the use of lefamulin remains limited in clinical practice. However, given the rise of antibiotic-resistant organisms, this may soon change. This review examines what is known about the pharmacokinetics and pharmacodynamics of lefamulin and looks ahead to its potential applications in clinical practice, including the treatment of sexually transmitted infections such as multidrug-resistant Mycoplasma genitalium, as well as its role as a synergistic agent used in combination with other antimicrobials in the treatment of drug-resistant organisms.

Introduction Haemophilus influenzae with a reduced susceptibility to quinolones (quinolone low-susceptible H. influenzae) has recently emerged in Japan. In addition, the regional outbreak of the quinolone low-susceptible H. influenzae ST422 clone has been reported. In this study, we isolated this clone from an acute care hospital located in a geographically different area from the previous outbreak and characterised the nature of this clone. Methods Eighty-nine H. influenzae isolated between 2017 and 2019 were tested. The antimicrobial susceptibility was determined by the broth dilution method. The genetic background was analysed by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing. Growth ability and β-lactamase acquisition were evaluated by growth curve analysis and conjugative transfer experiments, respectively. Results Quinolone low-susceptible isolates accounted for 4.2% (1/24) in 2018 and 13.9% (5/36) in 2019. Most of the quinolone low-susceptible strains (83.3%) were classified as ST422 and had amino acid substitutions in quinolone resistance-determining regions in both GyrA and ParC. The patients’ backgrounds were highly diverse. In addition, these isolates showed the same PFGE pattern as outbreak strains. The growth of ST422 clone was relatively faster than other clones. Furthermore, ST422 clone was able to acquire β-lactamase from a β-lactamase positive strain by horizontal transfer, becoming highly resistant to β-lactams. Conclusion Our study indicated that the quinolone low-susceptible H. influenzae ST422 clone has been spreading in the community undetected. In addition, this clone has the potential to grow faster and become more resistant through exogenous gene transfer. Therefore, ST422 clone should be monitored attention throughout Japan.

Abstract The review covers the study of the protective properties of IgA1 protease and the possibility of creating a vaccine preparation for the prevention of bacterial meningitis of various origins on its basis. Bacterial meningitis belongs to the group of socially dangerous diseases and is characterized by a severe course, numerous complications and high mortality. The approaches used at present in world practice to create antimicrobial vaccines are based on a narrow targeting against a specific pathogen. The development of a monocomponent vaccine against a wide range of bacterial pathogens with a common virulence factor is still relevant. IgA1 protease, a protein that is one of the main virulence factors of a number of gram-negative and gram-positive bacteria, can serve as such an antigen. Bacterial IgA1 protease is uniquely specific for immunoglobulins A1 (IgA1), cleaving peptide bonds in the hinge regions of the IgA1 in humans and other higher primates. Bacteria, getting on the mucous membrane, destroy IgA1, which acts as the first barrier to protect the body from infections. Neutralization of IgA1 protease at this stage can become an obstacle to the development of infection, hindering the adhesion of a number of pathogens that produce this protein. The data available in the literature on the mechanism of antibacterial protection are scattered and ambiguous. The review considers the literature data and the results of our own experiments on the protective activity of IgA1 protease. We have shown that the recombinant meningococcal IgA1 protease and some of its fragments protect mice from infection with a live virulent culture not only of meningococci of the main epidemic serogroups (A, B, C, and W135), but also of some of the most common virulent pneumococcal serotypes. The data obtained indicate the possibility of creating a monocomponent vaccine against these and, possibly, other bacterial infections. Currently, significant progress has been made in studying the structure and functions of secreted proteins in the bacteria Neisseria meningitidis and Haemophilus influenzae. In this review we describe protein translocation systems of N. meningitidis, which are related to the secretion of proteins in these bacteria, and also present modern data on the functions of these proteins. Analysis of experimental data on the structure of IgA1 protease of N. meningitidis and the formation of immunity during vaccination is of key importance in the development of prophylactic preparations.

Background Nasopharyngeal colonisation with nontypeable Haemophilus influenzae (NTHi) is associated with development of infections including pneumonia and otitis media. The 10-valent pneumococcal conjugate vaccine (PCV10) uses NTHi Protein D (PD) as a carrier. Papua New Guinean children have exceptionally early and dense NTHi carriage, and high rates of NTHi-associated disease. Vaccination with PCV10 could potentially reduce NTHi carriage and disease in this population by inducing a NTHi PD immune response. Methods Serum and nasopharyngeal swabs were collected from 101 Papua New Guinean children at 1, 4, 9, 10, 23 and 24 months of age. Children received PCV10 (n = 55) or PCV13 (not containing NTHi PD) (n = 46) at 1, 2 and 3 months of age. NTHi carriage density was measured in swabs by qPCR. Serum PD-IgG levels were measured by bead-based immunoassay. Results Papua New Guinean children did naturally develop PD-IgG antibodies whose levels were increased at 4 months of age with PCV10 vaccination at 1–2-3 months. Despite this, most children were colonised with NTHi by 4 months of age (~95%) regardless of being vaccinated with PCV10 or PCV13, and PCV10 had no impact on NTHi carriage density. Conclusion Early vaccination of infants with PCV10 elicited a robust PD antibody response but this had no impact on NTHi carriage. Trial registration ClinicalTrials.gov CTN NCT01619462.

Objectives To investigate the etiology of common respiratory pathogens in children < 2 years of age hospitalized with pneumonia in Xiamen from 2014 to 2017. Methods The medical records of 5581 children with pneumonia were retrospectively reviewed. Direct immunofluorescent test was used for respiratory virus testing. Bacteria were detected by conventional culture method. The results of pathogen detection at admission were analyzed as well as the clinical outcomes of children. Results The burden of hospitalized children with pneumonia was highest among infants < 6 months old (58.2%). Respiratory syncytial virus (RSV) was the most common respiratory virus (26.0%) followed by parainfluenza (4.8%) and adenovirus (3.2%). Haemophilus influenzae was the most common bacteria detected (16.6%) followed by Moraxella catarrhalis (13.4%), Staphylococcus aureus (13.0%), Streptococcus pneumoniae (12.3%), Escherichia coli (5.1%) and Klebsiella pneumoniae (4.8%). Notably, RSV and K. pneumoniae were detected more frequently in severe pneumonia (35.0% and 10.9%) versus mild pneumonia (25.6% and 4.6%), with higher rates of ICU admissions, longer hospital stays and higher hospital costs compared to those infected with other respiratory pathogens. Conclusions Among children < 2 years of age hospitalized with pneumonia in Xiamen, RSV was the most common respiratory virus, while H. influenzae and S. pneumoniae remained the predominant bacterial pathogens detected. Considering the low implementation rate of vaccines against pneumococcal and Hib pneumonia in China, there is an urgent need to increase both vaccination rates to reduce pneumococcal and Hib disease burden.

Respiratory infection is one of the infections that can be transmitted in closed places due to the ease of transmission of pathogens from infected people to healthy people through droplets from coughing and sneezing, among these bacterial causes, Haemophilus influenza. Therefore, this study aimed to evaluate the prevalence of this type of bacteria among pilgrims coming from different geographical locations in the world and determine the antibiotic-resistant strains. Around 1226 nasopharyngeal swabs were collected from 613 pilgrims from several different nationalities before and after completing the Hajj. These specimens were cultivated on chocolate agar medium and the pathogens were identified according to Clinical Laboratory Standards Institute (CLSI) protocol and confirmed by an automated system machine (VITEK2). There is no H. influenzae was detected among pilgrims before performing Hajj and the rate of transmission of H. influenzae was 1% after performing Hajj. The most effective antibiotics against to H. influenza isolates from pilgrims was Rifampicin while all the isolates were resistant to ampicillin. The continuous monitorization of the rate of potentially pathogenic bacterial during the Hajj season is required in order to develop advanced strategies to confront any outbreak of any infectious disease among the pilgrims. Key words: Hajj season, nasopharyngeal swabs, ethnic group, Makkah.

Background Community-acquired pneumonia (CAP) is one of the most common infectious diseases and is a significant cause of mortality and morbidity globally. A microbial cause was not determined in a sizable percentage of patients with CAP; there are increasing data to suggest regional differences in bacterial aetiology. We devised a multiplex real-time PCR assay for detecting four microorganisms (Streptococcus pneumoniae, Haemophilus influenzae, Klebsiella pneumoniae and Burkholderia pseudomallei) of relevance to CAP infections in Asia. Methods Analytical validation was accomplished using bacterial isolates (n=10–33 of each target organism for analytical sensitivity and n=117 for analytical sensitivity) and clinical validation using 58 culture-positive respiratory tract specimens. Results The qPCR assay exhibited 100% analytical sensitivity and analytical specificity, and 100% clinical sensitivity and 94–100% clinical specificity. The limit of detection and efficiency for the multiplex PCR assay were 3–33 CFU/mL and 93–110%, respectively. The results showed that the PCR-based method had higher sensitivity than traditional culture-based methods. The assay also demonstrated an ability to semiquantify bacterial loads. Conclusion We have devised a reliable laboratory-developed multiplex qPCR assay, with a turnaround time of within one working day, for detection of four clinically important CAP-associated microorganisms in Asia. The availability of a test with improved diagnostic capabilities potentially leads to an informed choice of antibiotic usage and appropriate management of the patient to achieve a better treatment outcome and financial savings.

Objectives/hypothesis Acute rhinosinusitis is a frequent common cold-related complication in children. Despite the need for appropriate treatment, its underlying microbiology remains unclear. This study aimed to investigate the microbiology of acute rhinosinusitis in children. Study design Prospective non controlled study. Methods Thirty-one pediatric acute maxillary sinusitis patients with severe symptoms were assessed. The subjects were 17 males and 14 females aged 5 to 14 years (mean age, 9.1 years). Maxillary sinus aspirates were collected and cultured, with subsequent viral and bacterial polymerase chain reaction (PCR) analysis. Bacteria were analyzed using culturing and PCR, and viruses were analyzed using PCR. The PCR kits used identify 18 types of respiratory viruses and 13 types of bacteria. Results At least one pathogen was detected in 30 of 31 aspirates (97%) using PCR, and none of the aspirates contained respiratory viruses alone. Ten aspirates (32%) contained both viruses and bacteria. The most common viruses detected were rhinovirus (13%) and influenza virus (10%). The most common bacteria were Haemophilus influenzae (45%), Streptococcus pneumoniae (32%), Moraxella catarrhalis (16%), and Chlamydophila pneumoniae (13%). Bacteria were found in 21 of 31 cases (68%) via bacterial culturing. Culturing revealed that H influenzae was the most common pathogen (42%). Conclusions In pediatric acute maxillary sinusitis, respiratory bacteria were detected in 65% of the sinus aspirates and both bacteria and viruses in 32%. The most common viruses were rhinovirus and influenza virus, and the most common bacteria were H influenzae and S pneumoniae. Viral and bacterial PCR is useful for accurately investigating the microbiology in pediatric sinusitis. Level of evidence Level 3. Laryngoscope, 2021.

Background Recurrent tonsillitis is one of the most common otolaryngological disorders caused by cell-invading bacteria, such as Streptococcus pyogenes (S. pyogenes) and Haemophilus influenzae. The aim of this study was to investigate the effect of antibacterial agents against cell-invading bacteria. Methods The intracellular invasion of Detroit 562 cells by five strains of nontypeable Haemophilus influenzae (NTHi) and four strains of S. pyogenes was investigated. The antibacterial agents used were garenoxacin (GRNX), clarithromycin (CAM), amoxicillin (AMPC), cefditoren pivoxil (CDTR-PI), and levofloxacin (LVFX). Results Both NTHi and S. pyogenes fully invaded Detroit 562 cells in 6 h and were less sensitive to CAM. GRNX, CAM, and LVFX were effective against bacteria invading the cells, but AMPC and CDTR-PI were not effective. GRNX was the most effective. Conclusion GRNX was the most effective agent against bacteria invading cells.

Carbohydrate-binding antibodies play diverse and critical roles in human health. Endogenous carbohydrate-binding antibodies that recognize bacterial, fungal, and other microbial carbohydrates prevent systemic infections and help maintain microbiome homeostasis. Anti-glycan antibodies can have both beneficial and detrimental effects. For example, alloantibodies to ABO blood group carbohydrates can help reduce the spread of some infectious diseases, but they also impose limitations for blood transfusions. Antibodies that recognize self-glycans can contribute to autoimmune diseases, such as Guillain-Barre syndrome. In addition to endogenous antibodies that arise through natural processes, a variety of vaccines induce anti-glycan antibodies as a primary mechanism of protection. Some examples of approved carbohydrate-based vaccines that have had a major impact on human health are against pneumococcus, Haemophilus influeanza type b, and Neisseria meningitidis. Monoclonal antibodies specifically targeting pathogen associated or tumor associated carbohydrate antigens (TACAs) are used clinically for both diagnostic and therapeutic purposes. This review aims to highlight some of the well-studied and critically important applications of anti-carbohydrate antibodies.

RESUMEN Desde un punto de vista microbiológico, tanto en el tratamiento antimicrobiano empírico como el dirigido en la infección respiratoria se fundamenta en el perfil de sensibilidad de los microrganismos aislados y los posibles mecanismos de resistencia que puedan presentar. Estos últimos puedan variar en diferentes áreas geográficas según los perfiles de prescripción y los programas de vacunación. Los antibióticos betalactámicos, las fluoroquinolonas y los macrólidos son los antimicrobianos más empleados en la exacerbación de la enfermedad pulmonar obstructiva crónica y la neumonía adquirida en la comunidad. En su prescripción se deben tener en cuenta aspectos como la actividad intrínseca, el efecto bactericida y su capacidad para evitar el desarrollo de resistencias. Este último está relacionado con los parámetros PK/PD, la concentración preventiva de mutantes y la denominada ventana de selección. Mas reciente ha crecido en importancia el potencial impacto ecológico, no solo sobre la microbiota intestinal, sino también sobre la respiratoria. El mantenimiento del estado de eubiosis requiere el uso de antimicrobianos con bajo perfil de actuación sobre los microorganismos anaerobios. Con ellos se facilita la resiliencia de las poblaciones bacterianas que integran la microbiota, el estado de resistencia de colonización y se limita el daño colateral relacionado con la emergencia de resistencia a los anti-microbianos en los patógenos que producen las infecciones y las poblaciones bacterianas que integran la microbiota.

The quality of culture media for blood culture was checked: nutrient medium for children with an antibiotic neutralizer for the cultivation of aerobes, nutrient medium with an antibiotic neutralizer for the cultivation of anaerobes, a nutrient medium with an antibiotic neutralizer for the cultivation of aerobes, nutrient medium for the cultivation of aerobes UNONA® used in the automatic bacteriological analyzer JUNONA ®Labstar 50 (SCENKER Biological Technology Co., Ltd. China). Used tenfold dilutions from 18-24 hour cultures of reference strains: ATCC 13124 Clostridium perfringens; ATCC 25285 Bacteroides fragilis; NCTC 194I8 Haemophilus influenzae; ATCC 49619 Streptococcus pneumoniae; ATCC 16615 Streptococcus pyogenes; ATCC 27853 Pseudomonas aeruginosa; ATCC 25923 Staphylococcus aureus; ATCC 25922 Escherichia coli; BKPGU-401/-885-653 Candida albicans; ATCC13813 Streptococcus agalactiae; No. 186 Enterobacter cloacae; ATCC 29212 Enterococcus faecalis; clinical isolates: Acinetobacter lwofii, Enterobacter cloacae, Candida tropicalis. All investigated reference strains were isolated on nutrient media in accordance with their biological properties when inoculated with 50 CFU / ml less than 72 hours later, as stated by the manufacturer. The study has shown that growth factors must be used to test the quality of the culture media with Haemophilus influenzae bacteria and this must be reflected in the manufacturer’s instructions.

Non-typeable Haemophilus influenzae (NTHi) is the most common respiratory pathogen in patients with chronic obstructive disease. Limited data is available investigating the impact of NTHi infections on cellular re-differentiation processes in the bronchial mucosa. The aim of this study was to assess the effects of stimulation with NTHi on the bronchial epithelium regarding cellular re-differentiation processes using primary bronchial epithelial cells harvested from infection-free patients undergoing bronchoscopy. The cells were then cultivated using an air-liquid interface and stimulated with NTHi and TGF-β. Markers of epithelial and mesenchymal cells were analyzed using immunofluorescence, Western blot and qRT-PCR. Stimulation with both NTHi and TGF-ß led to a marked increase in the expression of the mesenchymal marker vimentin, while E-cadherin as an epithelial marker maintained a stable expression throughout the experiments. Furthermore, expression of collagen 4 and the matrix-metallopeptidases 2 and 9 were increased after stimulation, while the expression of tissue inhibitors of metallopeptidases was not affected by pathogen stimulation. In this study we show a direct pathogen-induced trans-differentiation of primary bronchial epithelial cells resulting in a co-localization of epithelial and mesenchymal markers and an up-regulation of extracellular matrix components.

Background Haemophilus influenzae can cause invasive infections, in which cefotaxime is among the first-line antibiotics for treatment. The prevalence of cefotaxime-resistant H. influenzae in Europe is reported to be on a low level. Nevertheless, systematic studies with a large set of invasive isolates are scarce. Objectives To provide prevalence data for cefotaxime resistance in invasive H. influenzae isolates in Germany 2016–19 and investigate the epidemiological relevance of PBP3 mutations known to elevate the cefotaxime MIC. Methods Cefotaxime susceptibility of invasive H. influenzae isolates, collected in the national laboratory surveillance programme, was examined by gradient agar diffusion (GAD) testing. Cefotaxime resistance was determined according to EUCAST guidelines (resistance breakpoint MIC >0.125 mg/L). Therefore, the MIC for all resistant isolates was verified by broth microdilution method (BMD). WGS was performed to investigate the genetic relationship of cefotaxime-resistant isolates and to analyse alterations in the PBP3. An analysis of the geographic distribution of the resistant isolates was performed. Results From 2016 to 2019, the German National Reference Laboratory for Meningococci and H. influenzae received 2432 invasive H. influenzae isolates from blood and CSF. According to GAD results, 27 strains were resistant to cefotaxime. BMD confirmed the resistance in 22 of these isolates, which equals a prevalence of cefotaxime resistance of 0.90% in invasive H. influenzae in Germany. Among cefotaxime-resistant isolates cgMLST revealed three clusters. PBP3 analysis showed previously described mutations in our strains. In comparison with cefotaxime-susceptible strains, the alterations L389F and Y557H were significantly associated with cefotaxime resistance, but were not present in all resistant strains. Geographic analysis showed that the disease cases with cefotaxime-resistant H. influenzae were evenly spread throughout the population in Germany. Conclusions Cefotaxime is still well suited for the treatment of invasive H. influenzae infections. Rarely occurring cefotaxime resistance is caused by sporadic mutations. The role of PBP3 mutations needs further investigation.

Background We explored the nasal microbiota in Indigenous Australian children in relation to ear and nasal health. Methods In total, 103 Indigenous Australian children aged 2–7 years (mean 4.7 years) were recruited from 2 Queensland communities. Children’s ears, nose, and throats were examined and upper respiratory tract (URT) swabs collected. Clinical histories were obtained from parents/medical records. URT microbiota were characterized using culturomics with Matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) identification. Real-time PCR was used to quantify otopathogen (Haemophilus influenzae, Streptococcus pneumoniae, and Moraxella catarrhalis) loads and detect respiratory viruses. Data were analyzed using beta diversity measures, regression modeling, and a correlation network analysis. Results Children with historical/current otitis media (OM) or URT infection (URTI) had higher nasal otopathogen detection and loads and rhinovirus detection compared with healthy children (all P < .04). Children with purulent rhinorrhea had higher nasal otopathogen detection and loads and rhinovirus detection (P < .04) compared with healthy children. High otopathogen loads were correlated in children with historical/current OM or URTI, whereas Corynebacterium pseudodiphtheriticum and Dolosigranulum pigrum were correlated in healthy children. Conclusions Corynebacterium pseudodiphtheriticum and D. pigrum are associated with URT and ear health. The importance of the main otopathogens in URT disease/OM was confirmed, and their role relates to co-colonization and high otopathogens loads.

Haemophilus influenzae has contributed to key bacterial genome sequencing hallmarks, as being not only the first bacterium to be genome-sequenced, but also starring the first genome-wide analysis of chromosomes directly transformed with DNA from a divergent genotype, and pioneering Tn-seq methodologies. Over the years, the phenomenal and constantly evolving development of –omic technologies applied to a whole range of biological questions of clinical relevance in the H. influenzae-host interplay, has greatly moved forward our understanding of this human-adapted pathogen, responsible for multiple acute and chronic infections of the respiratory tract. In this way, essential genes, virulence factors, pathoadaptive traits, and multi-layer gene expression regulatory networks with both genomic and epigenomic complexity levels are being elucidated. Likewise, the unstoppable increasing whole genome sequencing information underpinning H. influenzae great genomic plasticity, mainly when referring to non-capsulated strains, poses major challenges to understand the genomic basis of clinically relevant phenotypes and even more, to clearly highlight potential targets of clinical interest for diagnostic, therapeutic or vaccine development. We review here how genomic, transcriptomic, proteomic and metabolomic-based approaches are great contributors to our current understanding of the interactions between H. influenzae and the human airways, and point possible strategies to maximize their usefulness in the context of biomedical research and clinical needs on this human-adapted bacterial pathogen., N.L.-L. is funded by a PhD studentship from Regional Navarra Govern, Spain, reference 0011-1408-2017-000000. C.G.-C. is funded by a PhD studentship from Agencia Española de Investigación (AEI), Spain, reference PRE2019-088382. This work has been funded by grants from MINECO RTI2018-096369-B-I00, 875/2019 from SEPAR, and PC150-151–152 from Gobierno de Navarra to J.G. CIBER is an initiative from Instituto de Salud Carlos III (ISCIII), Madrid, Spain.

Background Bacteria colonizing the upper respiratory tract (URT) of young children play a key role in the pathogenesis of lower respiratory tract infection (LRTI). Objectives To systematically review the literature on the association between bacteria colonizing the URT and LRTI among young children. Data sources MEDLINE, Academic Search Premier, Africa-Wide Information and CINAHL, Scopus and Web of Science. Study eligibility criteria Studies published between 1923 and 2020, investigating URT bacteria from LRTI cases and controls. Participants Children under 5 years with and without acute LRTI. Methods Three reviewers independently screened titles, abstracts and full texts. Meta-analysis was done using Mantel–Haenszel fixed- or random-effects models. Results Most eligible studies (41/50) tested nasopharyngeal specimens when investigating URT bacteria. Most studies were of cross-sectional design (44/50). Twenty-four studies were performed in children in lower- or lower-middle-income countries (LMICs). There was higher prevalence of Haemophilus influenzae (pooled OR 1.60; 95% CI 1.23–2.07) and Klebsiella spp. (pooled OR 2.04; 95% CI 1.17–3.55) from URT specimens of cases versus controls. We observed a positive association between the detection of Streptococcus pneumoniae from URT specimens and LRTI after excluding studies where there was more antibiotic treatment prior to sampling in cases vs. controls (pooled OR 1.41; 95% CI 1.04–1.90). High density colonization with S. pneumoniae (>6.9 log10 copies/mL) was associated with an increased risk for LRTI. The associations between both Streptococcus and Haemophilus URT detection and LRTI were supported, at genus level, by 16S rRNA sequencing. Evidence for the role of Moraxella catarrhalis and Staphylococcus aureus was inconclusive. Conclusions Detection of H. influenzae or Klebsiella spp. in the URT was associated with LRTI, while evidence for association with S. pneumoniae was less conclusive. Longitudinal studies assessing URT microbial communities, together with environmental and host factors are needed to better understand pathogenesis of childhood LRTI.

We investigated the nasopharyngeal microbiota in preschool patients hospitalized with lower respiratory tract infection to clarify the relationships between culturable nasopharyngeal bacteria and prognosis. From 2016 to 2018, nasopharyngeal culture was performed on inpatients under 6 years of age with a lower respiratory tract infection. Among the 1,056 study patients, 1,046 provided nasopharyngeal samples that yielded positive cultures, yielding 1,676 isolated strains. Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis, were isolated in 25%, 27%, and 31% of the samples, respectively, and were the major causes of respiratory tract infection in these children. The only factor associated with the isolation of antibiotic-resistant strains from the nasopharynx was daycare attendance, which did not affect clinical severity, such as duration of fever and hospitalization. This study demonstrated that resistant bacteria in the nasopharynx did not affect the severity of lower respiratory tract infection and supports the use of narrow-spectrum antimicrobial agents in accordance with published guidelines when initiating therapy for pediatric patients with community-acquired pneumonia.

Introduction: Despite the successful implementation of Haemophilus influenzae vaccination, invasive serotypes still lead to a fatal infection. We recently cared for a patient with a ventriculoperitoneal shunt (VPS) and H. influenzae meningitis and septicemia complicated by vasospasm. Vasospasm caused by Haemophilus central nervous system infection has not been previously reported. Case Presentation: A 34-month-old patient with a recent VPS presented with H. influenzae meningitis and sepsis. Despite the explant of hardware, followed by maximum medical management, the patient developed stroke due to severe vasospasm, which led to diffused anoxic brain injury. Conclusions: We aim to alert for the possible critical condition caused by H. influenzae. It is essential to treat the underlying illness, despite the presence of a VPS. Surgical implant tends to be overlooked by other subspecialists. Being vaccinated to H. influenzae does not protect from different subtypes like non-typeable H. influenzae. The cause of vasospasm remains unclear.

Metal and metal oxide nanoparticles (NPs) have been increasingly utilized in many industries to harness their documented antibacterial properties. However, the mechanism(s) of action is still debated in the literature. The aim of this study is to understand how changes in outer membrane charge of a test bacteria Haemophilus influenzae alter the antibacterial activity of ZnO NPs of average sizes of 20 nM and 60 nM. H. influenzae outer membrane charge was altered through use of the wild strain (Rd) and mutant lines H543 and H446. Results indicate that antibacterial effects are both concentration and size dependent, with smaller NPs causing increased antibacterial response. Most critically, antibacterial assays and collected TEM images demonstrate that increasing negative charge on the outer membrane of bacteria decreased the antibacterial activity of the ZnO NPs. Finally, this work demonstrates the possibility of using ZnO NPs to treat H. influenzae infection in clinical settings.

Over a 4-year study period from 2015 to 2018, altogether 183 isolates of bacterial meningitis were collected from 12 hospitals covering the entire Moroccan territory. Neisseria meningitidis represented 58.5%, Streptococcus pneumoniae 35.5%, and Haemophilus influenzae type b 6%. H. influenzae type b mainly affected 5-year-olds and unvaccinated adults. N. meningitidis serogroup B represented 90.7% followed by serogroup W135 with 6.5%. Decreased susceptibility to penicillin G (DSPG) for all isolates accounted for 15.7%, with 11.6% being resistant to penicillin G (PG) and 4.1% decreased susceptibility. Cumulative results of all strains showed 2.7% decreased susceptibility to amoxicillin and 3.3% resistant, 2.2% of isolates were resistant to third-generation cephalosporin and 2.2% were decreased susceptible, 5.5% were resistant to chloramphenicol and 2.7% were resistant to rifampin. The frequency of DSPG observed in our study is more common in S. pneumoniae than in N. meningitidis (P < 0.05). These isolates have been found to be highly susceptible to antibiotics used for treatment and prophylaxis chemotherapy and the observed resistance remains rare. The impact of introduction of conjugate vaccines against H. influenzae type b and S. pneumoniae (PCVs) is an advantage in reducing meningitis cases due to these two species.

Secretory otitis media (SOM) is characterized by persistence of fluid in the middle ear, often following an episode of acute otitis media. Our hypothesis is that failure to eliminate bacterial or viral pathogens may result in persistent low-grade inflammation. In this study, we analyzed inflammatory mediators in middle ear fluids from 67 children with SOM. This was combined with determinations of viable bacteria by culture along with detection of bacterial and viral genetic material by real-time polymerase chain reaction (PCR). The inflammatory mediators found at the highest concentrations (>30 ng/mL) were stem cell growth factor-β (median 110 ng/mL), CXCL1, IL-16, IL-8, migration inhibitory factor, CXCL10, and CXCL9. Among bacterial pathogens, Moraxella catarrhalis and Haemophilus influenzae dominated, regardless of detection methods, while rhinovirus dominated among viral pathogens. Middle ear fluid levels of interleukin (IL)-1α, IL-17, IL-1β, fibroblast growth factor basic, and tumor necrosis factor correlated strongly with presence of bacteria detected either by culture or PCR, while IL-1RA, IL-3, IL-6, IL-8, CCL3, CCL4, and granulocyte-colony stimulating factor correlated significantly with real-time PCR values. CXCL10, CXCL9, CCL2, and TRAIL correlated significantly with viral nucleic acid levels. To conclude, persistence of viral and bacterial pathogens may fuel persistent inflammation in SOM. Bacteria caused a broad inflammatory response, while viruses chiefly elicited the interferon-induced chemokines CXCL9 and CXCL10.

Community-acquired pneumonia (CAP) in children is still one of the leading causes of morbidity and mortality, especially in developing countries. The World Health Organization (WHO) has reported that pneumonia accounts for 15% of all deaths of children under 5 years old. The aim of the present study was to find out the predominance of microorganisms in the respiratory tract in children. 334 strains of microorganisms were isolated: Gram-positive – 293 strains, Gram-negative – 41. From the pharynx 183 strains were isolated, from the nose – 94, from sputum – 57. Among Gram-positive microorganisms the following were isolated: Staphylococcus aureus 44 strains of microorganisms, S. epidermidis – 75, Group A β-hemolytic streptococci – 39, viridans streptococci – 55, Streptococcus pneumoniae – 34, Enterococcus faecalis – 2, Candida spp. – 38, Corynebacterium pseudodiphthericum – 6. Among Gram-negative microorganisms the following were isolated: Escherichia coli 4 strains of microorganisms, Klebsiella pneumoniae – 13, Pseudomonas aeruginosa – 6, Haemophilus influenzae – 11, Enterobacter cloacae – 7. Children were divided by age and diagnosis into four groups: I group – children with acute bronchitis (0–5 years of age), II group – children with acute bronchitis (5–18 years of age), III group – children with CAP (0–5 years of age), IV group – children with CAP (5–18 years of age). Materials used in the research – nasal swabs, throat swabs and sputum. Microorganisms were isolated and identified using standard microbiological methods. S. aureus was the predominant microorganism isolate from the pharynx in children with bronchitis (0–5 years). Group A β-hemolytic streptococci were isolated most often from the pharynx in children with bronchitis older than 5 years and in children with CAP less than 5 years of age. S. pneumoniae was most often isolated from the pharynx in children older than 5 years of age with CAP. S. aureus was the main microorganism, isolated from the nose in children with bronchitis in all age groups; in patients with CAP it was the predominant microorganism in children older than 5 years of age. S. aureus was the predominant microorganism, isolated from sputum in children with bronchitis older than 5 years. S. pneumoniae was the predominant microorganism, isolated from sputum in children with CAP older than 5 years. The research showed that S. pneumoniae is still one of the main pathogens that cause CAP in school aged children.

Objectives To describe the characteristics of patients with a positive urethral sample for Haemophilus spp. Material and methods We performed a retrospective study from January 2018 to July 2019 at the Bordeaux university hospital (France) of all urethral samples positive for Haemophilus spp. Results Haemophilus spp. was isolated in 10 urethral samples from nine patients. The mean age was 33.8 years. Most patients reported having unprotected sex. Haemophilus parainfluenzae was isolated in nine samples, and Haemophilus influenzae in one sample. Antibiotic susceptibility tests were performed in five samples; Haemophilus spp. was always resistant to amoxicillin and tetracycline. One patient had persistent symptoms after treatment for a multidrug-resistant Haemophilus parainfluenzae strain. Conclusion Haemophilus spp. is a rare pathogen of urethritis. Its responsibility should be considered in case of persistent symptoms. The emergence of multidrug-resistant Haemophilus spp. is becoming problematic.

Background In the pediatric age group, approximately 7.5% of upper respiratory tract infections (URIs) are complicated by acute bacterial sinusitis (ABS). Despite its prevalence, ABS is often overlooked in young children. The diagnosis and management present unique challenges in primary care. This is an updated narrative review on the evaluation, diagnosis, and management of ABS. Methods A PubMed search was performed using the key term 'acute sinusitis'. The search strategy included clinical trials, meta-analyses, randomized controlled trials, observational studies, and reviews. The search was restricted to the English literature and children. Results Haemophilus influenzae (non-typeable), Streptococcus pneumoniae, and Moraxella catarrhalis are the major pathogens in uncomplicated ABS in otherwise healthy children. In complicated ABS, polymicrobial infections are common. The diagnosis of acute sinusitis is mainly clinical and based on stringent criteria, including persistent symptoms and signs of a URI beyond 10 days, without appreciable improvement; a URI with high fever and purulent nasal discharge at onset lasting for at least 3 consecutive days; and biphasic or worsening symptoms. Conclusion Data from high-quality studies on the management of ABS are limited. The present consensus is that amoxicillin-clavulanate, at a standard dose of 45 mg/kg/day orally, is the drug of choice for most cases of uncomplicated ABS in children in whom antibacterial resistance is not suspected. Alternatively, oral amoxicillin 90 mg/kg/day can be administered. For those with severe ABS or uncomplicated acute sinusitis who are at risk for severe disease or antibiotic resistance, oral high-dose amoxicillin-clavulanate (90 mg/kg/day) is the drug of choice.

Objectives: To identifyHaemophilus species and characterise the antimicrobial susceptibility of isolates from patients with respiratory tract infections (RTIs) in Cameroon. Methods: Isolates (n = 95) were from patients with RTIs obtained from two hospitals in Yaoundé, Cameroon. Isolates were identified by biochemical assay, a polymerase chain reaction (PCR)-based method, matrix-assisted laser desorption ionization-time of flight (MALDI-TOF), and whole genome sequencing. Antibiotic minimum inhibitory concentrations were determined by E-test. Results: Haemophilus influenzae (H. influenzae) was the most prevalent species, varying from 76.8 to 84.2% according to the different methods. The isolates were mainly non-typeable (n = 70, 96%). Three H. influenzae isolates were capsulated (b, e and f). The isolates were genetically diverse and 40 unique sequence types were identified, including 11 new ones. Resistance to ampicillin was observed among 52 of 94 (55.3%), and 14 of the 52 (26.9%) produced TEM-1 β-lactamase. PBP3 mutations occurred in 40 of 52 (76.9%) ampicillin-resistant isolates. Eleven isolates were chloramphenicol-resistant, with eight of 10 (80%) producing chloramphenicol acetyltransferase. Four Haemophilus isolates were rifampicin-resistant, with two mutations in rpoB gene. Five isolates were ciprofloxacin-resistant and harboured mutations in the quinolone-resistance-determining regions of gyrA and parC genes. Conclusion: TheH. influenzae isolates were highly diverse and showed high levels of antibiotic resistance. H. influenzae serotype b is still circulating in the post-vaccination era.

Background and aim Kaffir lime fruit peel oil and Kaffir lime leaf oil have been reported for their activities against respiratory tract pathogens. The purpose of the study was to develop clear oral sprays to be used as a first-defense oral spray. Experimental procedure Clear antibacterial oral sprays were prepared and analyzed for their respective active major compounds, using GC-MS. The sprays were tested against a Gr. A streptococcal clinical isolate and 3 standard respiratory tract pathogens, using Broth microdilution method. A 4-month stability test was carried out as well. Results and conclusion Six clear oral sprays, three formulae composed of Kaffir lime fruit peel oil (6, 10, 13%v/v KLO) and the other three formulae containing Kaffir lime leaf oil (4, 8, 12%v/v KLLO), were developed. The active compounds in KLO were α-terpineol and terpinene-4-ol whereas that in KLLO was citronellal. All oral sprays exhibited antibacterial activity against one Group A streptococcal clinical isolate and three respiratory pathogenic pathogens, Staphylococcus aureus ATCC 29213, Streptococcus pneumoniae ATCC 49619, and Haemophilus influenzae ATCC 49247, among which the strongest activity was against H. influenzae ATCC 49247. The antibacterial activity of all oral sprays remained unchanged in an accelerated stability test, at 4, 30, and 45 °C under 75% relative humidity, throughout the 4-month storage., Graphical abstract Image 101, Highlights of the findings and novelties • Clear antibacterial oral sprays were prepared. • Active major compounds were alpha-terpineol, terpinene-4-ol, and citronellal. • The sprays showed strongest activities against Haemophilus influenzae ATCC 49247. • The antibacterial activity was shown throughout 4-month storage.

Background Meningitis is a clinical syndrome, characterized by the inflammation of the meninges, the protective tissues that envelop the brain and spinal cord. It can be due to bacterial, viraland even fungal pathogens. Objectives This study’s aim is to investigate and observe the different aspects of meningitis in Lebanon between 2011 and 2019, specifically bacterial meningitis, and compare the incidence rates to different countries. Materials and methods This study is a descriptive epidemiological study. Data were collected from the epidemiological surveillance unit of the Lebanese Ministry of Public Health. Incidence according to year, months, age groups, governorates and causative agents were analyzed in the 9-years period. Results With an incidence rate of approximately 7.56 cases a year per 100,000 individuals, bacterial meningitis accounted for most of the meningitis cases from 2011 to 2019. Four of the most commonly identified bacterial pathogens were studied: Streptococcus pneumoniae, Neisseria meningitidis, Haemophilus influenzae, and Mycobacterium tuberculosis, as well as other cases of meningitis with an identified or unidentified agent. The most common identifiable pathogen was S. pneumoniae (13.06%), followed by N. meningitidis (7.09%), and H. influenzae (0.56%). Conclusion The data provided by the MOPH was evaluated in order to get a better understanding of the disease and improve the plans of action towards its control and treatment. Efforts should be made to preserve and ameliorate the existing surveillance system, and to ensure that all cases of meningitis are reported and investigated in compliance with the WHO guidelines.

Background/purpose Antimicrobial resistance in Taiwan has been on the rise for two decades. The implementation of pneumococcal conjugate vaccination (PCV13) and enhanced antimicrobial control (2013–2015) by the government may have changed the antibiotic resistance. Methods Four respiratory pathogens (Streptococcus pneumoniae, Haemophilus influenzae, Streptococcus pyogenes, and Moraxella catarrhalis) isolated in a single medical center during 2008–2017 were studied. We defined three temporal stages: (a) the first era (2008–2012), prior to implementation of the national immunization program (PCV13 vaccination), (b) the second era (2013–2015), during which an enhanced antibiotic control strategy was implemented, and (c) the third era (2016–2017), after implementation. Antimicrobial drug sensitivities were collected from two other hospitals: one from east Taiwan, one from west-central Taiwan. Results S. pneumoniae was frequently isolated during the first era. It declined progressively during the second era of PCV13 vaccination. S. pyogenes and M. catarrhalis were not frequently isolated. The drug susceptibility of S. pneumoniae to ceftriaxone and vancomycin remained high. The antimicrobial susceptibility of H. influenzae to amoxillin/clavulante declined over the three temporal stages, from 91.9%-79.5%–58.5% (all p Conclusion Antimicrobial resistance of H. influenzae increased during the latter part of the study period. The PCV13 vaccination program reduced the invasive pneumococcal disease and reduced the stress on the emergent drug resistance. This enhanced antibiotic control strategy was effective in terms of nosocomial drug resistance but not for community-associated pathogens.

X-linked agammaglobulinemia (XLA) is characterized by severe or recurrent infections, hypogammaglobulinemia, and circulating B cell deficiency. The frequent pathogens seen in patients with XLA include Streptococcus pneumoniae, Haemophilus influenzae, Pseudomonas aeruginosa, and enterovirus as well as Campylobacter and Helicobacter species. Here, we describe two patients with XLA who developed cellulitis and bacteremia caused by Helicobacter cinaedi even when administered an appropriate immunoglobulin replacement therapy. H. cinaedi may be difficult to isolate using a conventional blood culture system and could be identified by sequence analysis and mass spectrometry. H. cinaedi infection causes recurrent symptoms frequently, and patients require a long course of antibiotic treatment. Recently, the case of non-H. pylori Helicobacter (NHPH) infection such as H. cinaedi and H. bilis infection is increasing in number in patients with XLA. Systemic NHPH infection should be suspected, and extensive microbiological analysis should be performed to appropriately treat patients with XLA who present with fever and skin lesions.

Individuals with immunosuppressive condition have a high risk of invasive Haemophilus influenzae type b (Hib) infection. In Japan, routine Hib vaccination program for children under 5 years old was introduced in December 2008. However, the national policy does not make provision for individuals aged ≥5 years who have medical conditions associated with a high risk of invasive Hib disease to receive Hib vaccine. We measured serum anti-polyribosylribitol phosphate specific (anti-PRP) antibodies to Hib in patients aged ≥5 years with hematological malignancies and asplenia and evaluated their levels of anti-PRP antibodies in post administration of Hib vaccine era. A total of 65 patients (48 with hematological malignancies, and 17 with asplenia) were included in this study, of which 84% had not received Hib vaccine. In addition, 95.4% had short-term protective levels of anti-PRP antibodies (defined as ≥0.15 μg/mL) and 41.5% had long-term protective levels of anti-PRP antibodies (defined as ≥1.0 μg/mL). Five patients had low anti-PRP antibody levels despite a history of Hib vaccination. Our results suggest that young patients with underlying diseases such as hematological malignancies and asplenia may be at risk of invasive Hib disease. Hence, we recommend they should receive Hib vaccines even if they are over the age limit for routine Hib vaccination program.

Background: Emergence of resistance to some antibiotics in Haemophilus influenzae, a respiratory pathogen is a cause of concern. The aim is to study the antibiotic susceptibility pattern of Haemophilus isolates from respiratory infections with reference to beta-lactam resistance. Methods: This is a laboratory based prospective study done in the department of microbiology in a tertiary care center after institutional ethics committee clearance. Haemophilus influenzae isolates from respiratory tract specimens over a period of one year were subjected to antibiotic susceptibility tests. Beta-lactamase production was detected by nitrocefin disc. hpd gene, blaTEM and blaROB genes were detected by PCR. The data was analysed using SPSS 11.5 version. Results: Of the 162 isolates, 89.5% were from sputum specimens. Ampicillin resistance was seen in 5 (3.09%) isolates. The ampicillin resistant strains were positive for beta-lactamase enzyme and blaTEM gene. BLNAR and isolates with blaROB gene were not found. Conclusion: In case of Haemophilus influenzae respiratory tract infection empirical treatment with amoxicillin clavulanate or third generation cephalosporin may be the drugs of choice in our geographic area.

Background Otitis media (OM) is a common and potentially serious disease of childhood. Although OM is multifactorial on origin, bacterial infection is a unifying component. Many studies have established a critical role for innate immunity in bacterial clearance and OM resolution. A key component of innate immunity is the recruitment of immune and inflammatory cells, including macrophages. Methods To explore the role of macrophages in OM, we evaluated the expression of genes related to macrophage function during a complete episode of acute OM in the mouse caused by middle ear (ME) inoculation with Haemophilus influenzae. We also combined CCR2 deficiency with chlodronate liposome toxicity to deplete macrophages during OM. Results Macrophage genes were robustly regulated during OM. Moreover, macrophage depletion enhanced and prolonged the infiltration of neutrophils into the infected ME and increased the persistence of bacterial infection. Conclusions The results illustrate the critical role played by macrophages in OM resolution.

Acute Otitis Media (AOM) is a multifactorial disease occurring mostly in young children who are immunologically naïve to AOM pathogens. This review focuses on work from Rochester NY, USA over the past 12 years among young children who had AOM infections microbiologically-confirmed by tympanocentesis, so called “stringently-defined”. Among stringently-defined otitis prone children deficiencies in fundamental immune defense mechanisms have been identified that contribute to the propensity of young children to experience recurrent AOM. Dysfunction in innate immune responses that cause an immunopathological impact in the nasopharynx have been discovered including inadequate proinflammatory cytokine response and poor epithelial cell repair. Adaptive immunity defects in B cell function and immunologic memory resulting in low levels of antibody to otopathogen-specific antigens allows repeated infections. CD4+ and CD8+ T cell function and memory defects significantly contribute. The immune profile of an otitis prone child resembles that of a neonate through the first year of life. Immunologic deficits in otitis prone children cause them to be unusually vulnerable to viral upper respiratory infections and respond inadequately to routine pediatric vaccines.

Цель работы – изучение патогенетической роли Toll-подобных рецепторов-4 (ТПР-4) в развитии анемии воспаления.Материалы и методы. Изучили содержание TПР-4, ферритина и железа в сыворотке крови 78 детей (средний возраст – 1,5 ± 0,4 года) методом иммуноферментного анализа. В основную группу включили 46 детей с острыми воспалительными бактериальными заболеваниями органов дыхания, которых поделили на 2 подгруппы: первая – 24 ребенка с анемией воспаления, вторая – 22 ребенка без анемии. В зависимости от этиологического фактора основную группу исследования разделили на подгруппы: 1а – 12 пациентов с анемией воспаления, возбудитель – Streptococcus pneumoniae, 1b – 12 больных с анемией воспаления, возбудитель – Haemophilus influenzae, 2a – 11 детей без анемии воспаления, возбудитель – Streptococcus pneumoniae, 2b – 11 пациентов без анемии воспаления, возбудитель – Haemophilus influenzae. Группу сравнения составили 16 детей с железодефицитной анемией без воспалительных проявлений, контрольную группу – 16 условно здоровых детей. Группы наблюдения репрезентативны по возрасту и полу детей.Результаты. Установлено, что содержание ТПР-4 у детей в основной группе зависело от бактериального возбудителя. Содержание ТПР-4 в подгруппе, где этиологическим фактором был Haemophilus influenzae, превышало более чем 2 раза результаты группы контроля (p < 0,05), в 1,7 раза – группы сравнения, в 2 раза больше относительно их содержания в группе, где заболевание было обусловлено Streptococcus pneumoniae (p < 0,05). Отмечена тесная прямая корреляционная связь между содержанием ТПР-4 в группе пациентов, заболевание которых вызвано грамотрицательной флорой, и содержанием ферритина (r = 0,8, p < 0,05). Содержание железа в сыворотке крови детей с анемией воспаления было достоверно ниже показателей групп сравнения и контроля в 1,6 раза (8,78 (6,82–15,30) нг/мл и 13,88 (12,74–16,52) нг/мл соответственно, p < 0,05).Выводы. Развитие анемии воспаления у детей раннего возраста сопровождается повышением уровня ТПР-4, прежде всего в ответ на внедрение грамотрицательной микрофлоры (Haemophilus influenzae). Содержание ферритина находится в прямой зависимости от их уровня, что позволяет предположить пусковую роль ТПР-4в защитном механизме секвестрации железа, который является важнейшим звеном в патогенезе развития анемии воспаления., Aim. To determine the pathogenetic role of Toll-like receptors-4 (TLRs-4) in the development of anemia of inflammation (AI).Materials and methods. The content of TLRs-4, ferritin and iron in blood serum of 78 children (1.5 ± 0.4 years old on the average) was studied by enzyme-linked immunosorbent assay. The main group included 46 children with acute inflammatory bacterial diseases of the respiratory system, which were divided into 2 subgroups: the first subgroup included 24 children with AI, the second – 22 children without anemia. Depending on the etiological factor, the main group was divided into subgroups: 1a – 12 patients with AI, Streptococcus pneumoniae was detected as the pathogen, 1b – 12 patients with AI, the pathogen – Haemophilus influenzae, 2a – 11 patients without AI, the pathogen – Streptococcus pneumoniae, 2b – 11 patients without AI, the pathogen – Haemophilus influenzae. The comparison group included 16 children with iron deficiency anemia without inflammatory manifestations. Control group included 16 conditionally healthy children. The observation groups were representative by age and sex of the children.Results. It was determined that the content of TLRs-4 in children in the main group depended on the bacterial pathogen. The content of TLRs-4 in the subgroup where Haemophilus influenzae was the etiological factor exceeded more than 2 times the results of the control group (P < 0.05), 1.7 times the comparison group, and 2 times relative to their content in the group, where the disease was caused by Streptococcus pneumoniae (P < 0.05). There was a close correlation between the content of TLRs-4 in the group of patients whose disease was caused by gram-negative flora and the content of ferritin (r = 0.8, P < 0.05). The iron in the blood serum of children with anemia of inflammation was significantly lower than in the comparison and control groups by 1.6 times (8.78 (6.8215.30) ng/ml and 13.88 (12.74–16.52) ng/ml, respectively, P < 0.05).Conclusions. The development of AI in young children is accompanied by an increase in TLRs-4, primarily in response to the intrusion of gram-negative microflora (Haemophilus influenzae). The content of ferritin is directly dependent on their level, which suggests the starting role in the protective mechanism of iron sequestration, which is an important link in the pathogenesis of the development of AI., Мета роботи – вивчення патогенетичної ролі Toll-подібних рецепторів-4 (ТПР-4) у розвитку анемії запалення.Матеріали та методи. Вивчили вміст TПР-4, феритину й заліза в сироватці крові 78 дітей (середній вік – 1,5 ± 0,4 року) методом імуноферментного аналізу. В основну групу включили 46 дітей, які мали гострі запальні бактеріальні захворювання органів дихання; групу поділили на 2 підгрупи: перша – 24 дитини з анемією запалення, друга – 22 дитини без анемії. Залежно від етіологічного чинника основну групу дослідження поділили на підгрупи: 1а – 12 пацієнтів з анемією запалення, збудник – Streptococcus pneumoniae, 1b – 12 дітей з анемією запалення, збудник – Haemophilus influenzae, 2a – 11 хворих без анемії запалення, збудник – Streptococcus pneumoniae, 2b – 11 осіб без анемії запалення, збудник – Haemophilus influenzae. У групу порівняння залучили 16 дітей із залізодефіцитною анемією без запальних проявів, у контрольну групу – 16 умовно здорових дітей. Групи спостереження репрезентативні за віком і статтю дітей.Результати. Встановили, що вміст ТПР-4 в дітей основної групи залежав від бактеріального збудника. Вміст ТПР-4 в підгрупі, де етіологічним фактором була Haemophilus influenzae, більш ніж удвічі перевищував результати групи контролю (p < 0,05), в 1,7 раза – групи порівняння, вдвічі – щодо вмісту у групі, де захворювання зумовлено Streptococcus pneumoniae (p < 0,05). Встановили тісний прямий кореляційний зв’язок між концентрацією ТПР-4 у групі пацієнтів, захворювання яких спричинене грамнегативною флорою, та вмістом феритину (r = 0,8, p < 0,05). Вміст заліза в сироватці крові дітей з анемією запалення був вірогідно нижчим, ніж показники груп порівняння та контролю в 1,6 раза (8,78 (6,82–15,30) нг/мл і 13,88 (12,74–16,52) нг/мл відповідно, p < 0,05).Висновки. Розвиток анемії запалення в дітей раннього віку супроводжується підвищенням рівня ТПР-4, передусім у відповідь на грамнегативну мікрофлору (Haemophilus influenzae). Вміст феритину прямо залежить від їхнього рівня, що дає підстави для припущення щодо пускової ролі ТПР-4 в захисному механізмі секвестрації заліза, який є найважливішою ланкою в патогенезі розвитку анемії запалення.

Objectives: The aims of this study were to analyse the serotypes of epidemic Haemophilus influenzae and changes in mechanisms of β-lactam resistance over the past decade. Results: Haemophilus influenzae isolates in Western Sichuan from 2013–2014 were non-typeable H. influenzae (NTHi). β-Lactam MICs for NTHi isolated during 2013–2014 were significantly higher than those from 2003–2004 (P < 0.05). Of 274 NTHi, 141 (51.5%) were β-lactamase-positive (TEM-1 type). There were 35 amino acid (AA) substitutions in ftsI among NTHi isolated from 2013–2014. However, NTHi isolates from 2003–2004 had only nine AA substitutions. Ordered multiple classification logistic regression analysis showed that different AA substitution patterns in ftsI had different effects on β-lactam MICs. The main factors affecting the ampicillin MIC were the mutations R517H (OR = 6.999), L389F (OR = 7.128), N526K (OR = 4.660) and D350N (OR = 0.450). The main factor influencing the amoxicillin/clavulanic acid MIC was an N526K mutation (OR = 9.349). The main factors affecting the cefuroxime MIC were the mutations S357N (OR = 37.453) and N526K (OR = 14.816). Compared with 2003–2004, gBLNAR and gBLPAR isolated from 2013–2014 increased significantly from 13.0% (7/54) and 9.3% (5/54) to 38.2% (84/220) and 45.5% (100/220), respectively (P < 0.001). In the ‘others’ group of ftsI gene mutations, 13 NTHi had the same ftsI gene mutation pattern and 24 AA substitutions. Conclusion: These results confirm that β-lactam-resistant NTHi isolates increased rapidly. AA substitutions in ftsI were more complex and diversified in 2013–2014.

Phytochemicals from Capsicum anum L. plant extract are traditionally used to cure bronchitis. Bronchitis is caused by Haemophilus influenzae. Molecular docking method applied using “Biovia Discovery Studio”. “High positive values of -CDOCKER energy and -CDOCKER interaction energy” suggested that myrcetin and quercetin can effectively deactivate the Palmitoyl-CoA hydrolase enzyme thereby interrupting the life cycle of Haemophilus influenza.