Co-infection with Neisseria mucosa in a patient with tuberculous otitis media

Tuberculous otitis media(TOM) is a rare manifestation caused by Mycobacterium tuberculosis with low incidence rates among extrapulmonary tuberculosis cases. Diagnosis is often delayed because of the presence of several clinical manifestations and the high prevalence of secondary bacterial infections. Few reports have attributed secondary bacterial infections in patients with TOM to commensal Neisseria. Thus, understanding the pathogenic mechanisms and clinical features of commensal Neisseria is important, considering its recent presentation as an infection-causing pathogen. Neisseria mucosa is a commensal inhabitant in humans and is generally considered non-pathogenic but can cause infection in rare cases. Here, we report an atypical secondary infection caused by Neisseria mucosa in an 81-year-old woman with TOM being treated for pulmonary tuberculosis. Direct purulent otorrhea smear microscopy revealed no acid-fast bacilli using Ziehl-Neelsen staining, whereas the phagocytosis of gram-negative cocci by white blood cells was confirmed using Gram staining. Otorrhea culture revealed the growth of N. mucosa. Subsequently, M. tuberculosis infection in the otorrhea was identified using a culture-based method. Vigilance is critical for the early detection of TOM to prevent further complications. This report raises awareness regarding TOM and provides insight into the pathogenicity of N. mucosa in otitis media.

Neisseria mucosa-A rare cause of peritoneal dialysis-related peritonitis:A case report

BACKGROUND Neisseria mucosa is a gram negative diplococcus belonging to the genus Neisseria found commonly in the upper respiratory tract.It is typically a commensal organism when it is parasitic on oral and nasal mucosa.To our knowledge,it does not cause disease in healthy individuals with normal immunity,but can be pathogenic in those with impaired immune function or change in bacterial colonization site.Neisseria mucosa has been reported to cause bacterial meningitis,conjunctivitis,pneumonia,endocarditis,peritonitis and urethritis.However,peritoneal dialysis-related peritonitis caused by Neisseria mucosa is extremely rare in clinical practice,which has not previously been reported in China.CASE SUMMARY A 55-year-old female presented to the nephrology clinic with upper abdominal pain without apparent cause,accompanied by nausea,vomiting and diarrhea for two days.The patient had a history of Stage 5 chronic kidney disease for five years,combined with renal hypertension and renal anemia,and was treated with peritoneal dialysis for renal replacement therapy.The patient was subsequently diagnosed with peritoneal dialysis-related peritonitis.Routine examination of peritoneal dialysis fluid showed abdominal infection,and the results of microbial culture of the peritoneal dialysis fluid confirmed Neisseria mucosa.Imi-penem/cilastatin 1.0 g q12h was added to peritoneal dialysis fluid for anti-infection treatment.After 24 d,the patient underwent upper extremity arteriovenous fistulation.One month later,the patient was discharged home in a clinically stable state.CONCLUSION Peritonitis caused by Neisseria mucosa is rare.Patients with home-based self-dialysis cannot guarantee good medical and health conditions,and require education on self-protection.

Meningitis Outbreak Caused by Neisseria meningitidis Serogroup C ST 10217 in 2019 in Diapaga, Burkina Faso

Introduction: Meningitis caused by Neisseria meningitidis constitutes a burden for the countries in the meningitis belt of sub-Saharan in general and particularly for Burkina Faso. In 2019 the Diapaga health district experienced a meningitis epidemic due to N. meningitidis serogroup C. Methods: This is a cross-sectional study with a descriptive aim in the health district of Diapaga where all cases of meningitis were included in this work. Rapid diagnostic tests (RDTs), culture as well as real-time PCR were used for the biological analysis of cerebral spinal fluid (CSF) samples. Results: Of 155 CSF samples analysed, 42% (65/155) were tested positve. Of them, N. meningitidis C accounted for 83% of all positive cases. Likewise, all thirteen (13) NmC strains were susceptible to oxacillin, ceftriaxone, penicillin and chloramphenicol. All strains of NmC belonged to the sequence type (ST) 10 217 and to the clonal complex (CC) 10 217. These CCs belonged to the same variant PorA type: P1.21-15.16;FetA type: F1-7;PorB type: 3-463. Conclusion: Burkina Faso had known an epidemic of meningitis caused by NmC in 2019 in the health district of Diapaga. This outbreak was contained in time due to the performance of the epidemiological surveillance system which made it possible to investigate on time and introduce the vaccine against the pathogen NmC.

Prevalence and Risk Factors of Penicillinase-Type β-Lactamase Producing Neisseria gonorrhoeae Isolated from Patients Attending Health-Facilities in Yaounde, Cameroon

Background and Objectives: Mitigation of antibiotic resistant Neisseria gonorrhoeae has become a priority due to considerable health and economical disabilities it generates. In order to tackle the emergence of resistant Neisseria gonorrhoeae, this study aimed to determine the prevalence and risk factors of penicillinase type β-lactamase-producing Neisseria gonorrheae among patients consulting for genital infectious disorders in two health-facilities in Yaounde, Cameroon. Materials and Method: A cross-sectional descriptive and analytical study was conducted over a 3-month period, from July 2nd to October 2nd, 2022. Vaginal and urethral secretions were collected. Biochemical identification tests were performed on colonies grown on chocolate agar + polyvitex using the Api NH gallery. The detection of penicillinases was equally performed using the API NH gallery and confirmed using the antimicrobial susceptibility testing. The Minimum Inhibitory Concentrations of some antibiotics were determined using the E-Test. Results: The results showed that out of the 198 patients sampled, 16 (8.08%) were positive for Neisseria gonorrhoeae, among which 13/16 (81.25%) were penicillinase-type β-lactamase producers. Antimicrobial susceptibility testing results showed high co-resistances to antibiotics, mainly ciprofloxacin (100%), nalidixic acid (92.31%) and azithromycin (84.62%). Moreover, high Minimum Inhibitory Concentrations of ceftriaxone (ranging from 6 to 24 mg/L) was observed toward Neisseria gonorrhoeae isolates. The risk factors of the carriage of penicillinase-type β-lactamase producing Neisseria gonorrhoeae identified were: a history of Sexually Transmitted infections (p = 0.01) and unprotected sexual intercourse (p = 0.01). Conclusion: The emergence of penicillinase-type β-lactamase producing Neisseria gonorrhoeae is increasing and the situation is becoming worrisome. The identified risk factors can constitute a basic outlook to tackle resistant Neisseria gonorrhoeae, and therefore sustain antibiotic stewardship.

PCR Detection of Neisseria Gonorrhoeae Infection in Women by Self Obtained Low Vaginal Swabs

Objective: To evaluate the performance of polymerasechain reaction (PCR) using self obtained low vaginalswabs (SOLVS) to detect Neisseria gonorrhoeae (NG).Methods: One SOLVS and two cervical swabs werecollected from each of 298 female STD clinic attendees.PCR and culture were performed on both samples todetect NG.Results: Thirty-three cases of gonorrhoeae werediagnosed. The sensitivity of cervical culture, cervi-cal swab PCR and SOLVS PCR were 75.8% (25/33),87.9% (29/33) and 97.0% (32/33), respectively. Thespecificities of the respective methods were 100%(265/265), 99.6% (264/265) and 99.6% (264/265).The positive predictive values (PPVs) of the respectivemethods were 100% (25/25), 96.7% (29/30) and97.0% (32/33). The negative predictive values (NPVs)of the respective methods were 97.1% (265/273),98.5% (264/268) and 99.6% (264/265).Conclusions: The performance of SOLVS PCR is atleast as good as that of conventional cervical PCR tech-niques for the detection of NG. SOLVS may take theplace of cervical swabs for screening of NG infectionin women by PCR.

Detection of Neisseria Gonorrhoeae from Urine with Ligase Chain Reaction

Objective: To evaluate the value of ligase chainreaction(LCR) in the diagnosis of diplococcusgonorrhoeae in urine. Methods: LCR detection of the urine for Neisseriagonorrhoeae and bacteria culture of discharge was per-formed simultaneously to 276 patients with urethritisor cervicitis seeking treatment in sex transmitted dis-eases (STDs) outpatient clinic. For specimens withdiscordant results, polymerase chain reaction wasconducted. The purpose was to detect the respectivesensitivity and specificity of bacteria culture and LCR. Results: 24 of 276(8.7%) patients had positive LCRresults and 21 of 276(7.6%) were positive for culture.5 specimens had discordant results from LCR andbacteria culture. The sensitivity and specificity of LCRin the diagnosis of gonorrhoeae were 92.3% and100% respectively. Conclusion: This study showed that LCR had ahigher sensitivity and specificity for the diagnosis ofgonorrhoeae from urine.

Molecular Characteristics of Neisseria meningitidis Isolated during an Outbreak in a Jail: Association with the Spread and Distribution of ST-4821 Complex Serogroup C Clone in China

Objective To characterize the meningococcal strains isolated from cases and close contacts with meningococcal disease associated with an outbreak in a jail in May 2010 by investigating the national distribution of hyperinvasive ST-4821 serogroup C clone associated with this outbreak. Methods The cases were described based on the clinical symptoms and laboratory results. Pharyngeal swabs were cultured for N. meningitidis from men in the jail. Meningococcal isolates were identified by serogrouping, pulsed-field gel electrophoresis （PFGE） and multilocus sequence typing （MLST）, respectively. Four hundred and sixteen serogroup C N. meningitidis strains were collected from 27 provinces between 2003 and 2010 for a nationwide survey and analyzed by PFGE and MLST. Results Three persons in a jail system were infected with invasive N. meningitidis serogroup C. All isolates tested had matching PFGE patterns and belonged to the multilocus sequence type （ST） 4821 clonal complex. All 47 N. meningitidis strains were identified from the pharyngeal swabs of 166 peoples in the jail, and 26 of them belonged to ST-4821 serogroup C clone, and 90.14% （375/416） serogroup C strains identified in the nationwide survey belonged to the ST-4821 complex. The ST-4821 serogroup C clone was spread nationwide, distributed in 24 provinces, especially in eastern provinces between 2003 and 2010. Conclusion Endemic transmission and carriage rate of ST-4821 serogroup C clone are high in this jail system. The ST-4821 serogroup C clone is spreading in China and nationwide distributed despite the existence of some effective vaccines.

Typing of Neisseria gonorrhoeae Opa and NG-MAST Gene of 12 Pairs of Sexual Contact Gonorrhea Patients in China

To identify the genomic species of Neisseria gonorrhoeae, evaluate the difference between two molecular epidemiological methods and examine the relationship between sex partners and genotypes of bacteria, 24 strains of Neisseria gonorrhoeae isolated from the outpatients with gonorrhea were identified by using the Opa genotyping and NG-MAST genotyping and the relationship between genotypes and phenotypes was studied. Twenty-four strains of Neisseria gonorrhoeae fell into 10 ST genotypes by NG-MAST genotyping, whereas these strains were classified into 12 OT Opa genotypes by Opa genotyping. A new epidemic strain of ST genotype （217-86% homologisation 178） in China was identified. It is concluded that genotypes of each pair of strains from a pair of patient/sex partner besides 45/46 are the same, indicating that contagious infection take place between patient and the sex partner. Opa genotyping was more effective than NG-MAST genotyping in identifying the genomic species of Neisseria gonorrhoeae. ST genotype could be further classified into different Opa-types.

Fluoroquinolone Resistance and Mutation Patterns in gyrA and parC Genes in Neisseria gonorrhoeae Isolates from Shanghai, China

In order to study the resistance of Neisseria （N.） gonorrhoeae to the fluoroquinolone and detect mutation patterns of quinolone resistance-determining regions （QRDRs） of clinical isolates in Shanghai, China, a total of 80 clinical isolates of N. gonorrhoeae were consecutively collected from Shanghai. The MIC of fluoroquinolone for the isolates was examined by using the agar dilution method and the mutation profiles of the QRDRs of gyrA and parC were analyzed by sequencing and restriction fragment length polymorphism （RFLP）. Chi-square test was used for comparison of the t：nutation patterns. The results showed that： （1） High percentages of the 8 isolates were resistant to ciprofloxacin （95.0%）, ofloxacin （95.0%） and lomefloxacin （97.5%）, only one strain was susceptible to the ciprofloxacin. （2） Sensitive strains had a substitute of Asp95→Ala in the gyrA, and all isolates that were resistant or intermediated to the ciprofloxacin, had a double mutation in the gyrA （Ser91, Ala 92 and Asp95）. Some strains also had a mutation in the parC. （3） The MICs of these isolates were significantly associated with the mutation patterns in the gyrA and parC. A double mutation of gyrA combined with parC87 mutation was a predominant pattern in Shanghai and could mediate high level resistance to ciprofloxacin. It suggests that mutations in the QRDRs of gyrA and parC may be responsible for the fluoroquinolone resistance. And fluoroquinolone could not be used as the first line antibiotics for gonorrhea treatment any more in Shanghai, China.

Relationship between Mutation of IR in the mtr System of Neisseria Gonorrhoeae and Multiple Antibiotic Resistance

To study the relationship between mutation of the inverted repeat sequence （IR） in the multiple transferable resistant system （mtr） of Neisseria gonorrhoeae （NG） and its multiple antibiotic resistance, minimal inhibitory concentrations （MICs） for the clinically isolated strains were tested by agar-dilution-method. The mtr system＇s IR gene of NG was sequenced after amplification by polymerase chain reaction （PCR）. Either two susceptive or five penicillin-resistant strains had no base mutation in IR gene, while all of the 13 strains with multiple-antibiotic-resistance had a singlebase deletion （A/T）. The result suggests that a single-base deletion of the thirteen-base IR sequence in mtr system of NG might result in multiple antibiotic resistance but is not associated with single antibiotic resistance.

Antibiotic resistance of Neisseria species in Iran: A systematic review and meta-analysis

Objective: To estimate the prevalence of antibiotic resistance of Neisseria species in Iran. Methods: A systematic and electronic search using relevant keywords in major national and international databases was performed until 6th July, 2018 in order to find studies reporting the prevalence of antibiotic resistance of Neisseria species in Iran. Results: A total of nine studies were found to be eligible based on predefined inclusion and exclusion criteria. Our analysis indicated that the prevalence of Neisseria gonorrhoeae resistance to different antibiotics was as follows: 66.9% to penicillin, 59.1% to ciprofloxacin, 11.1% to ceftriaxone, 21.6% to spectinomycin, 13.8% to cefixime, 82.4% to co-trimoxazole, 52.7% to tetracycline, 29.9% to gentamicin, 87.5% to ampicillin, 11.1% to azithromycin, 2.2% to chloramphenicol, 50.1% to cefepime and 50.0% to vancomycin. Antimicrobial resistance rates of Neisseria meningitidis was as follows: 30.0% to penicillin, 33.3% to amoxicillin, 33.3% to cephalexin, 55.6% to ampicillin and 0.0% to ciprofloxacin, ceftriaxone, cefotaxime, amikacin, co-trimoxazole, gentamicin, kanamycin, chloramphenicol and ceftizoxime. Conclusion: Neisseria gonorrhoeae and Neisseria meningitidis isolates of Iran show resistance to different types of antibiotics. Therefore, care should be exercised for the use of penicillin, ciprofloxacin, co-trimoxazole, tetracycline, gentamicin, ampicillin, cefepime and vancomycin for gonococcal infections, and also with respect to the use of penicillin, amoxicillin, ampicillin and cephalexin for meningococcal infections in Iran.

Proteome Analysis of Neisseria meningitidis Serogroup Strains C Associated with Outbreaks in China

Objective During 2003-2005, an outbreak of meningitis due to Neisseria meningitidis serogroup C occurred in China. With the aim to find strain clues result in the final epidemics, the ancestral strain 053442, a clinical isolate, and a carrier strain 053426 with different gene type were analyzed. Methods Clinical strain 053442 and carrier strain 053426 were cultured on GC agar plates under the same condition. Two-dimensional electrophoresis was performed using the pH 3–10 nonlinear IPG strips of 24 cm length, and all the protein spots were identified by matrix-assisted laser desorption/ionization time of flight spectrometry. Results 502 and 380 protein spots were identified in 053426 and 053442 respectively, relating to 266 and 202 different genes covering a wide range of cellular functions. The express volume and number of proteins involved in energy metabolism, protein synthesis and amino acid biosynthesis in 053426 were higher than in 053442. Virulence factor Opa, Opc and a series of proteins involved in pilus assembly and retraction were identified in 053442, which appear to be of primary importance in colonization and invasion of human cells. Compared to 053442, virulence protein species were less in 053426, with lower express volumes too. No Opa and Opc were detected in 053426. Conclusion The different protein expression profiles of the clinical strain 053442 and carrier strain 053426 in the present study provide some clues of the different pathogenicity of the two strains, which may account for result in the final epidemics.

Construction of Prokaryotic Expression Plasmid of mtrC Protein of Neisseria gonorrhoeae and Its Expression in E.Coli

Summary： In order to provide a rational research basis for detection of resistance of Neisseria gonorrhoeae to antimierobial hydrophobie agents and study on the resistant mechanism of multiple transferable resistance （mtr） efflux system, plasmid pET-28a（＋） encoding mtrC gene was constructed and the related target protein was expressed in Escherichia colt （E. cold DE3. The fragments of mtrC gene of Neisseria gonorrhoeae from the standard strains were amplified and cloned into prokaryotic expression plasmid pET-28a（＋） with restriction endonuelease to construct recombinant pET-mtrC which was verified by restriction endonuelease and DNA sequencing. The recom- binant was transformed into E. coli DE3 to express the protein mtrC induced by IPTG. The results showed mtrC DNA fragment was proved correct through restriction endonuelease and DNA sequencing. hs sequence was 99.5 % homologus to that published on GeneBank （U14993）. A 48.5 kD fusion protein which was induced by IPTG was detected by SDS-PAGE. It was concluded that the construction of prokaryotic expression plasmid of mtrC protein of Neisseria gonorrhoeae was correct and the fusion protein was successively expressed in E. coli.

Study on Serotypes and Auxotypes of Neisseria Gonorrhoeae in Guangzhou

Objective:To investigate the serotypes and auxotypes distribution of Neisseria gonorrhoeae in Guangzhou. Method: 131 strains of Neisseria gonorrhoeae wereserotyped by co-agglutination test and 108 strains wereauxotyped by La Scolea's method. Results: Out of 131 strains of Neisseria gonorrhoeae,87.8% (115/131) were WⅡ/WⅢ, while 9.9% (13/131) wereWⅠ. The most important auxotypes were Proto, Pro and ILe,42.6% (46/108), 21.3% (23/108) and 12.0%, respectively. WⅡ/WⅢ was distributed among the all auxotypes aboveand WI found only in both Proto and Pro. Conclusion: The study illustrated the prevailing serotype,WⅡ/WⅢ, and higher prevalence of Ile- in Guangzhou.

Cloning and expression of Neisseria meningitides luxS gene

Neisseria meningitides is a Gram-negative bacterium which is an important causative agent of septicemia and meningitis.Numerous pathogenic bacteria contain luxS,which is required for autoinducer-2 production.Neisseria meningitides contains a functional copy of luxS that is necessary for full meningococcal virulence.Neisseria meningitides DNA was extracted and its luxS gene was amplified by nested PCR.PCR product was purified and cloned in to pQE-30 expression vector.Recombinant plasmid was transformed and mass cultured.luxS was expressed in E.coli and confirmed by western blot analysis.In this study,N.meningitides luxS was amplified, cloned and expressed successfully.The sequencing of PCR product confirmed that amplified gene was luxS. Gene was expressed and observed in SDS-PAGE.Protein was reacted by his tag monoclonal antibody through western blot analysis.

Excellent outcome of primary Neisseria meningitidis keratoconjunctivitis

Infectious conjunctivitis is a very common presentation to medical professional and ophthalmologist all over the world.Although its typically self-limiting and treatable in almost all of the cases,but we need to be aware of the rare and potentially life threatening if the cause is not promptly identified and treated accordingly.In our case report,we highlighted the rare case of Neisseria meningitidis as a primary cause of keratoconjunctivitis.Neisseria meningitidis is a rare etiology of keratoconjunctivitis and its ocular presentations are quite similar with other bacterial or viral infection.The infection may potentially fatal if systemic invasion occurred,however with immediate and proper treatment the outcome is satisfactory.Early diagnosis and proper antibiotic treatment are critical to prevent systemic spread of the infection.Public health intervention is needed to prevent outbreak of the disease.

Construction of Prokaryotic Expression Plasmid of Fusion Protein Including Porin A and Porin B of Neisseria Gonorrhoeae and Its Expression in E.coli

In order to provide a rational research basis for clinical detection and genetic engineering vaccine, plasmid pET-28a (+) encoding both Porin gene PIA and PIB of Neisseria gonorrhoeae was constructed and a fusion protein in E.coli DE3 expressed. The fragments of PIA and PIB gene of Neisseria gonorrhoeae were amplified and cloned into prokaryotic expression plasmid pET-28a(+) with double restriction endonuclease cut to construct recombinant pET-PIB-PIA. The recombinant was verified with restriction endonuclease and sequenced and transformed into E.coli DE3 to express the fusion protein PIB-PIA after induced with IPTG. The results showed PIA-PIB fusion DNA fragment was proved correct through sequencing. A 67 kD (1 kD=0 992 1 ku) fusion protein had been detected by SDS-PAGE. It was concluded that the fusion protein was successively expressed.

Effect of Atmospheric Pressure Non-equilibrium Plasmas on Neisseria gonorrhoeae

In this study, the sterilizing effect of atmospheric pressure nonequilibrium plasmas （APNPs） on Neisseria gonorrhoeae （N. gonorrhoeae） was preliminarily examined and the possible mechanisms were explored. N. gonorrhoeae FA1090, FA19 and MSll were treated by APNPs and their survival rate was analyzed by using CFUs counting and structurally studied by laser scanning confocal microscopy. The morphological changes of bacterial cell membrane and wall were studied under TEM. Our results showed that APNPs had strong sterilizing effect on N. gonorrhoeae. The survival rate of MS11 in N. gonorrhoeae liquid medium was 60.65% after disinfection with the APNPs for 5 rain, whereas, the survival rate of FA19 was 92.60% and the rate of FA1090 was 96.40%. The survival rate of MS 11 was 21.13% after exposure to APNPs for 6 rain, whereas the survival rate of FA19 was 31.60% and the rate of FA1090 was 91.00%. N. gonorrhoeae was structurally damaged after treatment with APNPs. It is concluded that APNPs is able to effectively and quickly kill the N. gonorrhoeae, and the killing effect is related to the architectural damage of cell membrane.

Construtcion of Neisseria Gonorrhoeae Porin B Plasmid Recombinant and Its Expression in E.coli

A prokaryotic expression recombinant plasmid pET-PIB to express porin B (PIB) of Neisseria gonorrhoeae in E.coli DE_3 was constructed in order to provide a basis of research in detection, prophylactic and therapeutic vaccine against the pathogen infection. The gene encoding PIB was amplified by PCR from Neisseria gonorrhoeae and cloned into prokaryotic expression plasmid pET-28a(+) to construct a pET-PIB recombinant, which was verified by restriction endonuclease and DNA sequencing. Protein PIB was expressed in E.coli DE_3 induced with IPTG. The antigenicity of the expressed protein was evaluated by indirect ELISA. Rabbits were immunized with the protein and serum was collected after immunization. To assess the immunogenicity of the protein, the titer of serum to protein PIB was determined by ELISA. DNA sequence analysis showed that the nucleic acid sequence of PIB gene was 99.28 % of homology compared with that (NGPIB18) published in GenBank. A 41 kD fused protein was detected by SDS-PAGE and was proven to have reactivity with anti-PIB polyclonal antibody from mouse. A polyclonal antibody to PIB of 1:4000 titer determined by indirect ELISA was obtained from rabbit immunized with the purified product. Recombinant plasmid encoding PIB of Neisseria gonorrhoeae was constructed. Protein PIB with antigenicity and immunogenicity was successfully expressed.

In vitro Recombination and Identification of Mutated Fragment Corresponding to Regulation Region of mtrR Gene of Neisseria Gonorrhoeae

A site-directed mutant DNA fragment Neisseria Gonorrhoeae （NG） stains to construct the was synthesized and transfected into clinical transformants that contained the corresponding mutagenesis of regulation region of mtrR gene. According to the technique of gene splicing by overlap extension （SOEing）, a DNA segment with specific mutagenesis was constructed by two-step polymerase chain reaction （PCR）. The mutation fragments EF could be used for the next experiment in which the mutation NG strains were induced. By comparing the recombinant EF fragments to the corresponding DNA fragments of clinical NG strains, 2 of these were not compatible completely. The results of sequencing revealed that there was a 9 bp deletion between the 45 to 54 inverted repeat sequence localized within the mtrR promoter. It can be confirmed that the fragments EF are the specifically designed mutant fragments.