Protective eff ect and mechanism of nanoantimicrobial peptide ND-C14 against Streptococcus pneumoniae infection

BACKGROUND:Streptococcus pneumoniae(S.pneumoniae)is a common pathogen that causes bacterial pneumonia.However,with increasing bacterial resistance,there is an urgent need to develop new drugs to treat S.pneumoniae infections.Nanodefensin with a 14-carbon saturated fatty acid(ND-C14)is a novel nanoantimicrobial peptide designed by modifying myristic acid at the C-terminus of humanα-defensin 5(HD5)via an amide bond.However,it is unclear whether ND-C14 is effective against lung infections caused by S.pneumoniae.METHODS:In vitro,three groups were established,including the control group,and the HD5 and ND-C14 treatment groups.A virtual colony-count assay was used to evaluate the antibacterial activity of HD5 and ND-C14 against S.pneumoniae.The morphological changes of S.pneumoniae treated with HD5 or ND-C14 were observed by scanning electron microscopy.In vivo,mice were divided into sham,vehicle,and ND-C14 treatment groups.Mice in the sham group were treated with 25μL of phosphate-buffered saline(PBS).Mice in the vehicle and ND-C14 treatment groups were treated with intratracheal instillation of 25μL of bacterial suspension with 2×108 CFU/mL(total bacterial count:5×10^(6) CFU),and then the mice were given 25μL PBS or intratracheally injected with 25μL of ND-C14(including 20μg or 50μg),respectively.Survival rates were evaluated in the vehicle and ND-C14 treatment groups.Bacterial burden in the blood and bronchoalveolar lavage fluid were counted.The lung histology of the mice was assessed.A propidium iodide uptake assay was used to clarify the destructive eff ect of ND-C14 against S.pneumoniae.RESULTS:Compared with HD5,ND-C14 had a better bactericidal eff ect against S.pneumoniae because of its stronger ability to destroy the membrane structure of S.pneumoniae in vitro.In vivo,ND-C14 significantly delayed the death time and improved the survival rate of mice infected with S.pneumoniae.ND-C14 reduced bacterial burden and lung tissue injury.Moreover,ND-C14 had a membrane permeation eff ect on S.pneumoniae,and its destructive ability increased with increasing ND-C14 concentration.CONCLUSION:The ND-C14 may improve bactericidal eff ects on S.pneumoniae both in vitro and in vivo.

Acute coalescent mastoiditis in a 16-month-old child due to Streptococcus pneumoniae infection

Rationale:Acute otitis media is a common disease in early childhood,and is usually caused by Streptococcus pneumoniae(S.pneumoniae).Acute mastoiditis is a complication of acute otitis media and can involve not only the mucoperiosteum of the middle ear but can also spread to the periosteum by destroying the mastoid bone(acute coalescent mastoiditis).In addition,the infection can extend through the surrounding bones or the emissary veins beyond the mastoid’s air cells,leading to subperiosteal abscesses.Patient’s Concern:A 16-month-old female patient was hospitalized due to the purulent discharge of the left ear and the symptoms of right mastoiditis(swelling and redness of the skin).Diagnosis:Bilateral acute coalescent mastoiditis caused by S.pneumoniae infection.The computer tomography revealed bilateral bone destruction of the mastoid and abscesses found behind the auricle on both sides.Interventions:The patient underwent intravenous antibiotic therapy and surgical treatment.Outcomes:The patient was discharged 14 days after hospitalization with an improved condition.Lessons:Improperly treated acute coalescent mastoiditis can lead to extracranial and intracranial complications,sometimes serious and even life-threatening.Complications are prevalent in children under 2 years,in whom the disease progresses more rapidly and severely.The vaccination with a 13-valent vaccine may not result in sufficient immunity against S.pneumoniae,a predominant pathogen in children affected by acute coalescent mastoiditis.

Antibiotic Resistance Profile of Serotypes of Streptococcus pneumoniae Strains in Bangui, from 2017 to 2022: Case of Serotype 1

Goals: The aim of this study was to determine the antibiotic resistance profile of serotypes of Streptococcus pneumoniae strains circulating in Bangui. Methodology: A prospective and analytical analysis was carried out at the National Laboratory of Clinical Biology and Public Health from 2017 to 2022. The strains came from our study on the contribution to the study of antibiotic sensitivity of Streptococcus pneumoniae strains. The multiplex PCR test was used for its cost-effectiveness in terms of amplifiers which can be purified in order to be sequenced. It also makes it possible to detect several germs as well as their serotypes. For a PCR reaction, several elements are involved in the reaction medium or Master Mix. These are the desoxyribonucleotides (dNTPs), the magnesium ions (MgCl2) and the primers. A set of 14 primers divided into 3 classes were used. Class 1 primers served as an internal control by targeting the cpsA gene. It is a highly conserved gene found in capsular loci characterized to date. The primers of the second class were used to target specific serotypes by specific reactions (out of six possibilities). The group reaction was carried out using the primers of the third class in order to carry out an initial screening of the samples and to classify the pneumococcal isolates. Related serotypes were grouped based on the amplification of common genes. Using the technique of electrophoresis on agarose gel and an ultraviolet radiation device, the migration bands are then visualized and analyzed. The data collected had been entered into Excel 2010 and analyzed with Epi info 7. The exact Fischer chi2 test at the 5% threshold, the relative risk and its 95% confidence interval were used to compare the proportions and determine the associations. Results: 187 antibiotic-resistant strains of Streptococcus pneumoniae were collected. The average frequency of serotypes 1, 9A, 4 and untypeable identified were 43.59%, 18.18%, 18.27% and 39.57% respectively. The frequency of serotype 1 was predominant for the age group over five years old with 56.88%. The male sex was predominant with 55.08% for serotype 1. Resistance to penicillin and gentamicin for serotype 1 during this study, for the age group under 5 years old, was 77%. For serotypes 19A and 4, tetracycline resistance was predominant with 20% for the age group under 5 years. The resistance to penicillin and gentamicin of non-typeable serotypes was 33% for the age group under 5 years old. For the age group over 5 years old, resistance to erythromycin predominated at 37%. The distribution of serotypes by sex depending on antibiotic resistance was variable. There was a statistically significant association between identified serotypes and antibiotic resistance (p Conclusion: The study determined serotypes 1, serotypes 19A, serotypes 4 and non-typeable serotypes. These results would be due to the quality of vaccination or poor protection of vaccines.

A Cross-sectional Survey Assessing Carriage of Streptococcus pneumoniae in a Healthy Population in Xinjiang Uygur Autonomous Region of China

The carriage rate and serotype distribution of Streptococcus pneumoniae（S.pneumoniae）in a healthy population in China remains unclear.In this study,we collected the oropharyngeal swabs from513 individuals in Xinjiang,China.Real-time PCR targeting the lytA gene and 12 serotypes were assessed to identify S.pneumoniae carriage.

Antimicrobial susceptibility patterns of Streptococcus pneumoniae over 6 years at Gondar University Hospital, Northwest Ethiopia

Objective:To assess the magnitude and antimicrobial susceptibility patterns ofStreptococcus pneumoniaeisolates from various clinical specimens.Methods:A record based on retrospectivestudy was conducted at Gondar University Teaching Hospital from September 2007 to January 2012.All patients who visited Gondar University Hospital and provided clinical specimens(body fluids,discharge,swab and blood)for routine bacteriological culturing and antimicrobial susceptibilitytesting were taken for analysis.Clinical specimens were processed for bacterial culture accordingto the standard procedures.Antimicrobial susceptibility test for isolated organisms was doneusing agar disk diffusion method.The data were entered and analyzed using SPSS software version16 package.Results:One hundred and fifty threeStreptococcus pneumoniaewere isolated frompatients who visited Gondar University Teaching Hospital bacteriology laboratory for culture.Majority of the pneumococcal isolates were from inpatients[111(72.5%)],and 74(48.4%)were frombody fluids.Out of the total isolates,93(61%)were found to be resistant to at least one antibioticused for susceptibility testing.Forty eight(43.2%)of the isolates were multi-drug resistant(resistantto two or more drugs).The resistance rate noted for both ciprofloxacin 17(11.1%)and ceftriaxone15(9.8%)were alarming.Conclusions:High proportions of the isolates tend to be increasinglyresistant to the commonly prescribed drugs.The recommended drug of choice like ciprofloxacinand ceftriaxone were found to be less susceptible in the study area.Based on the findings,wetherefore recommend that antimicrobial agents should be inspected for acceptable activity beforethey are prescribed and administered empirically.Further study with a better design and surveyof antimicrobial susceptibility at large scale shoule be performed to draw advanced information.

Children with Pneumonia Caused by Streptococcus pneumoniae Resistance Analysis and Clinical Features

Objective: To analyze the causes of childhood pneumonia Streptococcus pneumoniae resistance and clinical characteristics, and provide a basis for better and timely clinical therapy, and medication to reduce blind-ness. Methods: MIC method in our hospital 114 under 2020 pediatric pneumococcal respiratory infection in children with lower respiratory tract specimens were isolated antimicrobial susceptibility testing, and analyzed retrospectively. Results: 84 male children, 30 female children, the largest of which 9 years old, the youngest two months, infants less than 1 year old, 90 people;suffering from bronchial pneumonia, 90 cases, 21 cases of pneumonia, wheezing, 3 cases of bronchitis, the average length of stay for about a week;improved in 79 cases, 33 cases were cured, 2 cases transferred to higher level hospitals. All children with throat congestion, swollen tonsils, lung breath sounds rough, smell and moist rales. 114 penicillin-resistant Streptococcus pneumoniae was 64.9%, erythromycin 97.4%, clindamycin 86.8%, tetracycline 87.7%, trimethoprim-sulfamethoxazole 82.5%, amoxicillin 21.9%, cefotaxime 49.1%, chloramphenicol 10.5%, was not found to levofloxacin and van-comycin. Conclusion: Penicillin, erythromycin, and clindamycin are not as pneumococcal pneumonia in children experience preferred medi-cation in children less than one year old child could easily cause lung chain Streptococcus pneumoniae. Therefore, the antimicrobial resistance of Streptococcus pneumoniae analysis provides a reference for experi-enced clinicians to adjust medication.

Expression,Purification,Characteristics and Homology Modeling of the HMGS from Streptococcus pneumoniae

Objective To understand the molecular basis for a potential reaction mechanism and develop novel antibiotics with homology modeling for 3-hydroxy-3-methylglutaryl coenzyme A （HMG-CoA） synthase （HMGS）. Methods The genetic engineering technology and the composer module of SYBYL7.0 program were used, while the HMGS three-dimensional structure was analyzed by homology modeling. Results The mvaS gene was cloned from Streptococcus pneumoniae and overexpressed in Escherichia coli from a pET28 vector. The expressed enzyme （about 46 kDa） was purified by affinity chromatography with a specific activity of 3.24 μmol/min/mg. Optimal conditions were pH 9.75 and 10 mmol/L MgCl2 at 37 ℃ The Vmax and Km were 4.69 μmol/min/mg and 213 μmol/L respectively. The 3D model of S.pneumoniae HMGS was established based on structure template of HMGS of Enterococcus faecalis. Conelusion The structure of HMGS will facilitate the structure-based design of alternative drugs to cholesterol-lowering therapies or to novel antibiotics to the Gram-positive cocci, whereas the recombinant HMGS will prove useful for drug development against a different enzyme in the mevalonate pathway.

Impact of the Pneumococcal Heptavalent Conjugated Vaccine on <i>Streptococcus pneumoniae</i>Nasopharyngeal Carriage and Antimicrobial Susceptibility in Children 2-5-Year-Old in Beijing, China

Streptococcus pneumoniae is a primary cause of illness and death among children younger than 5 years in China. The heptavalent pneumococcal conjugate vaccine (PCV7) was the only conjugated vaccine (PCV) available in China from 2008 to 2013. This randomized, controlled, open-label study conducted at 46 Beijing clinics involved 3281 healthy 2-5-year-old Chinese children, randomized 1:1 to receive one dose of the S. pneumoniae heptavalent conjugated vaccine (PCV7) (n = 1643) or Haemophilus influenzae type b conjugate vaccine (Hib) (n = 1638). The main objective of this study was to investigate the impact of PCV7 against that of Hib vaccination in the nasopharyngeal S. pneumoniae colonization in healthy Chinese children. Nasopharyngeal (NP) samples for culture, serotyping and antimicrobial susceptibility testing were collected before vaccination and at Day 60 and 180 post-vaccination. A total 3281 children were enrolled in the study. Demographic characteristics were similar among both study groups: 1641 children received PCV7. Before immunization, S. pneumoniae was isolated in 338 and 360 children in the PCV7 (144 PCV7 isolates) and Hib groups (145 PCV7 isolates), respectively. At Day 180, PCV7 vaccination was more effective than Hib vaccination in reduction NP carriage (20.2% [P = 0.052]) and new acquisition (19.0% [P = 0.066]). When reductions in NP carriage and new acquisition of PCV7 VT plus 6A was analyzed, reduction in the PCV7 vaccinated group achieved statistical significance (P = 0.034 and P = 0.042 versus Hib, respectively). NP carriage of NVT increased in both groups (P = 0.305 between study groups at Day 180). PCV7 decreased NP carriage of non-susceptible VT to amoxicillin (P = 0.000), ceftriaxone (P = 0.047) and MDR (P = 0.024) versus Hib. PCV7 vaccination in Chinese children 2 to 5 years of age was more effective than vaccination with Hib in the reduction of S. pneumoniae nasopharyngeal carriage, new acquisition and non-susceptible isolates.

Prevalence and Antimicrobial Susceptibility Pattern of Streptococcus pneumoniae at General Hospital in the Central Region of Japan from December 2013 to February 2014

Streptococcus pneumonia infection is important cause of morbidity and mortality. This study was conducted to find out the prevalence and antimicrobial susceptibility pattern of Streptococcus pneumoniae isolates at general hospitalin the central region of Japan from December 2013 to February 2014. Streptococcus pneumoniae was identified by standard laboratory procedure. Antimicrobial susceptibility testing was performed by micro dilution assay according to CLSI recommendation. One hundred fifty-three Streptococcus pneumoniae were isolated among which 80 (52.2%) were males and 73 (47.8%) were females. Nasal discharge (134%/87.6%) contributed more than other biological materials. The age incidence of (0 - 1) years, (1 - 10) years, (11 - 40) years, (41 - 60) years and >60 years age groups were 26 (17.0%), 110 (71.9%), 3 (2.0%), 10 (6.5%), and 4 (2.6%) respectively. Positive samples were received mostly from the pediatrics (137%/89.5%), respiratory medicine (12%/7.8%) and lowest from gastroenterology (1%/0.6%) and neurology (1/ 0.6%) department. Vancomycin and rifampicin were the most active antibiotics with 100% susceptibility rates. The next best were levofloxacin, penicillin G and ceftriaxone. Our study revealed that 82 Streptococcus pneumonia isolates had multidrug resistant ability (53.6%). Streptococcus pneumoniae infection spreads among community easily and inappropriate use of antibiotics contributes to their resistance. Continuous antimicrobial susceptible surveys are essential to guide policy on the adequate use of antibiotics to reduce the morbidity and mortality and reduce the emergency of antimicrobial resistance.

<i>Streptococcus pneumoniae</i>Upper Respiratory Carriage in Costa Rican Children with Otitis Media before the Introduction of the Heptavalent Conjugated Vaccine in the National Immunization Program

Objective: The aim of this study was to analyze the NP/OP S. pneumoniae serotype distribution and potential vaccine coverage in Costa Rican children with Otitis Media (OM) before the introduction of PCV-7 in the National Immunization Program (NIP). Methods: Between 2002 and 2006, NP and OP samples were obtained from 641 children from 6 to 79 months of age, at the time of OM diagnosis. S. pneumoniae serotyping and antimicrobial susceptibility were performed. Results: 386 S. pneumoniae isolates were recovered. The most common S. pneumoniae serotypes (ST) were: ST 6B, ST 14, ST 19F. Penicillin non-susceptibility was observed among 57% of the isolates obtained from children < 24 months of age. 15% strains were multidrug resistant. Potential vaccine coverage was: PCV-7: 60%;PCV-10: 62%;and PCV-13: 76% and against penicillin non-susceptible and multidrug resistant isolates was: PCV-7;59% and 83%, respectively;PCV-10: 60% and 85%, respectively and PCV-13: 74% and 96%, respectively. Conclusions: S. pneumoniae was isolated from the NP and/or OP in the majority (59%) of studied children with OM. At a statistical significant level, only serotype 3 was more frequently isolated among children >24 months of age. Antibiotic non-susceptibility and MDR were significantly higher in children <24 months of age. This study demonstrates that PCV-13 offers the highest potential vaccine coverage and serves to assess the impact of introduction of one of the conjugated vaccines in the NIP in Costa Rica.

Wrist Septic Arthritis Due to <i>Streptococcus pneumoniae</i>Revealing a Selective Immunoglobulin G4 Subclass Deficiciency: A Case Report

The pneumococcal septic arthritis is uncommon in healthy adults. In most cases, it occurs in patients with immune deficiency. Its exceptional location at the wrist can maintain doubt with rheumatic causes which are more frequent. IgG4 deficiency found in this patient was the only cause that could be impugned. We report the case of a young woman of 46 years without any evident cause of immunodeficiency and any particular risk factor, managed for septic arthritis of the wrist due to Streptococcus pneumoniae. This case without any apparent risk factors demonstrates that pneumococcal infection in a more distant articulation of the ENT is still possible. Scan has been a very significant contribution in confirming the positive diagnosis that leads to surgical drainage. The approach of the systematic search for the gateway must be done. In the study of the patient’s immune status, the realization of the protein profile could be interesting and then guide the practitioner in the decision to vaccinate the patient against Streptococcus pneumoniae if he was not. We recommend looking always for a cause of immunodeficiency including primary Ig defiencies even among adult.

Evolution of Erythromycin Resistance among Streptococcus pneumoniae Isolates in Malaysia from 2005 and 2010

Objectives: This study focuses on the antibiotic susceptibility pattern and distribution of the ermB and mefA virulence genes among the Streptococcus pneumoniae due to an increase in erythromycin resistance in S. pneumoniae worldwide. Methodology: We investigated 255 clinical isolates collected from 2005-2010 to determine the serotype distribution and resistance to erythromycin in comparison to penicillin, clindamycin, clarithromycin, azithromycin, and trimethoprim sulfamethoxazole. Multiplex PCR was carried out to detect erythromycin resistance genes (ermB and mefA). Results: There were 146 (57.3%) isolates resistant to erythromycin. MIC90 for erythromycin is at >256 mg/L and MIC50 is at 16 mg/L. The ermB gene was detected in 25.3% of the erythromycin-resistant isolates and mefA gene was detected in 50.7% of the isolates. The four most common serotypes encountered are 19F, 19A, 23F and 14. The serotype distribution among the erythromycin resistant isolates was 19F (42.0%) followed by serotype 19A (11.3%), serotype 23F (9.2%) and serogroup 14 (7.0%). Conclusion: In conclusion, there is a significant rise in erythromycin resistance among the Malaysian pneumococcal isolates. The emergence of serotype 19A together with increasing prevalence of resistance to macrolide warrants for a more extensive surveillance study.

Emergence of Optochin Resistance among <i>Streptococcus pneumoniae</i>in Japan

The optochin susceptibility test is a key method for differentiating Streptococcus pneumoniae from other α-hemolytic streptococci;however, optochin-resistant (Optr) S. pneumoniae have been reported in the last two decades. In this study, we investigated the isolation frequency of Optr S. pneumoniae in the North Kyushu area of Japan, and biochemically and genetically characterized Optr S. pneumoniae clinical isolates. Seven (0.68%) out of 1032 S. pneumoniae isolates collected by the North Fukuoka Infectious Diseases Working Group were found to be Optr S. pneumoniae. Resistant strains had MICs of optochin 2- to 64- fold higher than susceptible strains, possessed different antimicrobial resistance profiles, and belonged to different serotypes. All the seven Optr isolates had mutations in the nucleotide sequence code for subunit c of F0F1 ATPase. Three isolates had mutations in codon 48 (deduced amino acid substitution of valine with phenylalanine) and two isolates had mutations in codon 49 (substitution of alanine with threonine or serine). Of the remaining two isolates, one had mutation in codon 50 (substitution of phenylalanine with leucine) and the other had mutation in codon 44 (substitution of methionine with isoleucine, which was a novel mutation in this position). From these results, we identified the mutation in the H+-ATPase subunit c gene (atpC) of S. pneumoniae, which was not recognized earlier, and determined that Opt resistance among Japanese pneumococcal isolates is not related to a specific pneumococcal serotype or antimicrobial resistance profile. Furthermore, the results indicate that when α-hemolytic streptococci resistant to optochin are isolated from patients with invasive infectious diseases, such as meningitis and pneumonia, we should perform additional examinations such as bile solubility tests or PCR assays before confirming isolates as viridans streptococci. This is the first report of the characterization of Optr S. pneumoniae in Japan.

Streptococcus pneumoniae and Herpes Simplex Virus-1 Central Nervous System Co-Infection

Co-infections of the central nervous system (CNS) caused by bacterial and viral pathogens are considered to be rare. Herpes simplex virus type-1 (HSV-1) reactivation following Streptococcus pneumoniae infection is well described but most cases are related to oral or cutaneous lesions or in respiratory samples. HSV-1 CNS reactivation after Streptococcus pneumoniae meningitis is a very rare event and may have significant morbidity and mortality. In this case report, we describe a 71-year-old female patient that presented with a history of abdominal pain and confusion/disorientation that had tonic-clonic seizures while in the Emergency Department. The diagnostic work-up confirmed CNS co-infection caused by Streptococcus pneumoniae and HSV-1. Of note, beyond age, the patient had no known risk factors for both entities and recovered fully after antibiotic and antiviral therapy. This case underlines that clinicians must be aware of CNS co-infection despite being a rare diagnosis. This should be suspected particularly in patients who present an unusual clinical course of CNS infection.

Contribution to the Study of Antibiotic Sensitivity of Streptococcus pneumoniae Strains in Spinal Cerebral Fluids in Bangui from 2017 to 2022

A prospective and analytical study was carried out from January 2017 to Decem-ber 2022, at the National Laboratory of Clinical Biology and Public Health (LNBCSP) in Bangui. 352 samples of cerebrospinal fluid (CSF) were confirmed out of 2065, coming from the four hospitals in Bangui. This study aimed to evalu-ate the evolution of antibiotic sensitivity to strains of Streptococcus pneumoniae. CSF had been collected from patients who presented with meningeal syndromes. Based on the leukocyte count (White blood cells ≥ 5 mm3), an aliquot was trans-ferred to trans-isolate medium and sent to the LNBCSP for confirmatory testing, culture and determination of antibiotic sensitivity. The antibiotic sensitivity of Streptococcus pneumoniae strains was tested according to the methods recom-mended by the Antibiogram Committee of the French Society of Microbiology. The data collected was entered into Excel 2010 to be analyzed with Epi Info 7.2. Ficher’s exact test, chi2 at the 5% threshold (p < 0.05) was used to compare pro-portions and analyze associations between variables. The average sensitivity rate to β-lactams was 74.43%. The sensitivity rate of Fluoroquinolones was 54.54%. That of levofloxacin was 87%. The average rate of sensitivity to β-lactams for the age group under 5 years old was 79.25%. That of fluoroquinolones was 52.59%. Levofloxacin had 90.37%. The average sensitivity rate to β-lactams for the age group over 5 years old was 76.03%. Fluoroquinolones had 45.16%. Levofloxacin had 69.58%. The average sensitivity rate to β-lactams for males was 76.68%. Fluoroquinolones had 54.26%. That of levofloxacin was 83.40%. The sensitivity rate to β-lactams for females was 74.41%. That of fluoroquinolones was 51.16%. Levofloxacin had 67.44%. Cyclins had 28.68%. The study noted an association between age and sensitivity (p < 0.05;CI [1.05-2.57]). Strains of Streptococcus pneumoniae were always detected in the CSF. The average rate of sensitivity to macrolides was 36.93%;aminoglycosides 28.69%;phenicols 63.35%;sulfona-mides 39.2%. These results could suggest a reduced sensitivity to β lactams. .

Antimicrobial susceptibility and serotypes of Neisseria meningitidis and Streptococcus pneumoniae in Sri Lanka:Experience from the National Reference Laboratory

Objective:To determine the antimicrobial susceptibility and serotypes of Neisseria(N.)meningitidis and Streptococcus(S.)pneumoniae in Sri Lankan patients.Methods:We retrospectively analyzed 11 blood culture specimens from suspected patients with invasive meningococcal disease and 26 S.pneumoniae clinical isolates.We tested 6 antimicrobials against N.meningitidis and 12 antimicrobials against S.pneumoniae.Meningococcal serogroup was determined by realtime PCR and Quellung serotyping was used for pneumococcal analysis.Results:N.meningitidis serogroup B was the most common in this study.Intermediate-susceptibility to penicillin was seen in 75.0%(6/8)of strains.Susceptibility to ciprofloxacin,levofloxacin and cotrimoxazole was 62.5%(5/8),62.5%(5/8)and 87.5%(7/8),respectively.Excellent susceptibility was seen in cefotaxime and meropenem.In S.pneumoniae,the most common serotype was 19F in both invasive and non-invasive pneumococcal diseases.The majority of strains showed multidrug resistance.Penicillin non-susceptibility in non-meningeal strains were 13.6%and all meningeal strains were penicillin resistant.Erythromycin was highly resistant in both groups.Amoxicillin showed excellent susceptibility in non-invasive pneumococcal diseases strains.Linezolid,levofloxacin and vancomycin showed 100.0%susceptibility in all pneumococcal isolates.Conclusions:Implementation of vaccines should be considered,especially for children and high-risk populations.This may contribute to reducing pneumococcal and meningococcal invasive disease burden and help prevent emergence of antimicrobial resistant strains.

Development of protective antibodies to Streptococcus pneumoniae in healthy children

Aim: To study how immunity to Streptococcus pneumoniae normally develops in healthy children. Methods: Ninety two healthy children at 3 - 5, 7 - 9 and 13 - 15 years of age were recruited. No one of the children had previously been given pneumococcal vaccine. Serum was analysed for pooled antigens of the 23 most common pneumococcal polysaccharides with ELISA technique, and results are given in opitical density (OD). A three-level semi-quantitative system was used to assess degree of immunity to Streptococcus pneumoniae . Cut-off levels were OD ≤ 0.7 and OD ≥ 1.3, separating low, intermediate and high degree of immunity. Results: Median values for OD differed significantly between the groups, with OD 0.91, 1.18 and 1.10 respectively (p = 0.004). Levels were lower in the youngest age group, but from age 7-9 years, levels were similar trough out childhood. Twenty six percent of the children in the youngest age-group had a low degree of protection (OD ≤ 0.7) to Streptococcus pneumoniae . Such low levels were uncommon from 7 - 9 years of age, found in only 13% of the children. Conclusion: Protective antibodies to Streptococcus pneumoniae develops mainly during the preschool period. Thereafter, levels are stable throughout childhood up to the age of 15 years.

Penicillin Resistance in Nasopharyngeal Streptococcus pneumoniae among Children with Sickle Cell Disease Immunized with 7-Valent Pneumococcal Conjugate Vaccine

To assess the prevalence, the antimicrobials resistance and to identify risk factors of nasopharyngeal colonization by Streptococcus pneumoniae in children with sickle cell disease (SCD) using prophylactic penicillin and immunized with 7-valent pneumococcal conjugate vaccine. A total of 424 swabs were collected from the nasopharynx in 216 children with SCD and 109 samples from the control group, both from 2 to 60 months age range. Isolation and identification of pneumococci followed standard procedures. Minimum inhibitory concentration (MIC) for penicillin was determined by the E-test method. Prevalence of nasopharyngeal colonization by pneumococci in children with SCD was 17%, and 11% in the control group. The risk factors for increased colonization in children with SCD were the presence of more than five people at home, daycare/school attendance and low prophylaxis compliance. The prevalence of strains with penicillin resistance was 57.5% in patients with SCD and 25% in the control group. Conclusions: Prevalence of pneumococci nasopharyngeal colonization was similar among the study groups. Prophylactic use of penicillin may have increased the prevalence of resistant strains. The vaccine did not decrease the colonization with penicillin resistant strains.

Klebsiella pneumoniae infections after liver transplantation:Drug resistance and distribution of pathogens,risk factors,and influence on outcomes

BACKGROUND Liver transplantation(LT)is the only curative treatment for end-stage liver disease.However,LT recipients are susceptible to infection,which is the leading cause of early mortality after LT.Klebsiella pneumoniae infections(KPIs)in the bloodstream are common in LT recipients.We hypothesized that KPIs and carbapenemresistant Klebsiella pneumoniae(CRKP)infections may affect the outcomes of LT recipients.AIM To assess KPI incidence,timing,distribution,drug resistance,and risk factors following LT and its association with outcomes.METHODS This retrospective study included 406 patients undergoing LT at The Third Xiangya Hospital of Central South University,a tertiary hospital,from January 2015 to January 2023.We investigated the risk factors for KPIs and assessed the impact of KPIs and CRKP infections on the prognosis of LT recipients using logistic regression analysis.RESULTS KPI incidence was 7.9%(n=32),with lung/thoracic cavity the most frequent site of infection;the median time from LT to KPI onset was 7.5 d.Of 44 Klebsiella pneumoniae isolates,43(97.7%)and 34(77.3%)were susceptible to polymyxin B or ceftazidime/avibactam and tigecycline,respectively;>70%were resistant to piperacillin/tazobactam,ceftazidime,cefepime,aztreonam,meropenem,and levofloxacin.Female sex[odds ratio(OR)=2.827,95%confidence interval(CI):1.256-6.364;P=0.012],pre-LT diabetes(OR=2.794,95%CI:1.070-7.294;P=0.036),day 1 post-LT alanine aminotransferase(ALT)levels≥1500 U/L(OR=3.645,95%CI:1.671-7.950;P=0.001),and post-LT urethral catheter duration over 4 d(OR=2.266,95%CI:1.016-5.054;P=0.046)were risk factors for KPI.CRKP infections,but not KPIs,were risk factors for 6-month all-cause mortality post-LT.CONCLUSION KPIs occur frequently and rapidly after LT.Risk factors include female sex,pre-LT diabetes,increased post-LT ALT levels,and urethral catheter duration.CRKP infections,and not KPIs,affect mortality.

表达Strep-tag II的重组塞内卡病毒的构建与鉴定

为构建表达Strep-tag II的重组塞内卡病毒(SVA),本研究在感染性cDNA克隆p SVA-I212V/S460L的基础上,采用重叠延伸PCR扩增Strep-tag II并采用无缝克隆法将其插入SVA VP1基因C端,构建感染性cDNA克隆,命名为p SVA-Strep,并采用双酶切和测序鉴定正确后,将p SVA-Strep转染BHK-21细胞拯救重组病毒,收集病变细胞上清,并将含有病毒的细胞上清和野生型SVA(SVA-WT)分别感染BHK-21细胞,以未感染病毒的BHK-21细胞作为阴性对照,感染后24 h分别以鼠源SVA VP2蛋白单克隆抗体(MAb)5D10(1∶1000)、鼠源Strep-tag II MAb(1∶2000)作为一抗,以FITC标记的山羊抗小鼠Ig G(1∶2000)为二抗,采用间接免疫荧光试验(IFA)对重组病毒鉴定。IFA结果显示,以鼠源SVA VP2蛋白MAb 5D10为一抗,SVA-WT和重组病毒感染的细胞均出现绿色荧光,以Strep-tag II MAb作为一抗时,仅有重组病毒感染的细胞出现绿色荧光,经SVA-WT感染的BHK-21细胞及阴性对照细胞均无绿色荧光,表明拯救了Strep-tag II标记的重组病毒,并将其命名为r SVA-Strep。将r SVA-Strep在BHK-21细胞中连续传10代,每隔2代采用引物VP1-F/R经PCR鉴定并对第10代重组病毒的VP1基因测序以鉴定r SVA-Strep的遗传稳定性。结果显示,每隔两代的重组病毒经PCR扩增后均出现约780 bp的目的条带,第10代重组病毒扩增的VP1基因测序结果显示Strep-tag II基因仍在重组病毒中,且无基因突变,表明r SVA-Strep的遗传稳定性较强;将SVA-WT和r SVA-Strep分别以MOI 0.01感染BHK-21细胞,在感染后4 h、8 h、12 h、24 h、36 h、48 h和72 h收获病毒测定病毒滴度并绘制生长曲线,结果显示在病毒感染36 h内r SVA-Strep和SVA-WT生长动力学基本一致,表明Strep-tag II的引入在病毒感染36 h内对SVA的复制基本无影响。采用0.2%甲醛充分灭活r SVA-Strep,并利用Strep-TactinRXT Purification纯化试剂盒纯化该灭活病毒,将r SVA-Strep灭活病毒上清加至Strep-Tactin亲和层析柱,分别用不同体积洗脱缓冲液依次洗脱病毒并通过western blot鉴定病毒的纯化效果。对纯化过程中不同洗脱液洗脱病毒的western blot结果显示,不同体积的洗脱液中E1、E2和E3均出现VP0和VP2两条带,且E1和E2的条带最明显,表明洗脱缓冲液的最佳洗脱体积约为2.2 CV;以上结果表明r SVA-Strep可通过Strep-Tactin亲和层析柱一步法纯化。本研究在SVA中插入Strep-tag II标签,为SVA灭活疫苗的快速纯化提供技术手段,也为其他病原的快速纯化方法的建立提供参考。